Zlib - Pub Practical Skills in Chemistry

Practical Skills in Chemistry John R. Dean • Alan M.
Jones • David Holmes

John R. Dean • Alan M. Jones • David Holmes
Rob Reed • Jonathan Weyers • Allan Jones Rob Reed • Jonathan Weyers • Allan Jones
If you are studying chemistry, or a chemistry-related course, then this book will be an
indispensable companion throughout your entire degree programme. This ‘one-stop’ text will
guide you through the wide range of practical, analytical and data-handling skills that you will
need during your studies. It will also give you a solid grounding in wider transferable skills such Practical Skills in
Chemistry
as teamwork, using information technology, communicating information and study skills.
Chemistry
Practical Skills in
Now in its third edition, Practical Skills in Chemistry, 3e, has been enhanced and updated
throughout to provide a complete and easy-to-read guide to the developing skills required
from your first day through to graduation, further strengthening its reputation as the practical
resource for students of chemistry and related discipline areas.
This edition provides:

• New content layout to aid readability.
• ‘One-stop’ guide to the key practical skills needed in chemistry. THIRD EDITION
• Information presented in a clear and user friendly manner, tailored directly for the mastery of
basic laboratory skills.
• Numerous margin tips and hints, ‘how to’ boxes, checklists, worked examples and study
exercises.
• Guidance on numerical aspects, including statistics.
• Chapters grouped into key topics.
• Fully updated guide to health and safety, project work, Raman spectroscopy, internet
resources and enhancing your CV.
• New chapters on molecular formulae and X-ray diffraction.
• Generic guidance regarding Microsoft Office software, rather than version-specific details.
Practical Skills in Chemistry, 3rd edition, is an indispensable book for undergraduate

students in chemistry and related discipline areas and a useful primer for post-graduate
students. It is also a valuable resource for teachers in secondary schools.
THIRD
John R. Dean is Professor in Analytical and Environmental Sciences at the University of
Northumbria at Newcastle.
EDITION
Alan M. Jones was Head of Chemical Sciences at the University of Northumbria at Newcastle.
Reed • Weyers • Jones

Dean • Jones • Holmes
David Holmes is Associate Dean of the Keith B. Taylor Global Scholars Programme,
St George’s University School of Medicine (Grenada), based at the University of Northumbria at
Newcastle.
Rob Reed is Adjunct Professor (Education & Science) at Central Queensland University,
Australia.
Jonathan Weyers is Honorary Senior Lecturer at the University of Dundee.
Allan Jones is the Chancellor’s Award Fellow in Ecology, Environmental Science and Zoology
at the University of Dundee.
Cover image: Michael Rozewski/Getty Images www.pearson-books.com
CVR_DEAN_03_39920.indd 1 02/05/2017 14:01

Practical Skills in Chemistry
A01 Practical Skills in Chemistry 39920.indd 1 11/05/2017 09:13

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Practical Skills in Chemistry
Third Edition
John R. Dean
Alan M. Jones
David Holmes
Rob Reed
Jonathan Weyers
Allan Jones
Harlow, England • London • New York • Boston • San Francisco • Toronto • Sydney • Dubai • Singapore • Hong Kong
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First published 2002 (print)

Second edition published 2011 (print and electronic)
Third edition published 2017 (print and electronic)
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© Pearson Education Limited 2011, 2017 (print and electronic)
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to be identified as authors of this work have been asserted by them in accordance with the Copyright,
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ISBN: 978-1-292-13992-0 (print)

978-1-292-13993-7 (PDF)
978-1-292-13994-4 (ePub)
British Library Cataloguing-in-Publication Data

A catalogue record for the print edition is available from the British Library
Library of Congress Cataloging-in-Publication Data

Names: Dean, John R.
Title: Practical skills in chemistry.
Description: Third edition / John R. Dean [and five others]. | Harlow,
England : Pearson, 2017. | Includes bibliographical references and index.
Identifiers: LCCN 2017002095| ISBN 9781292139920 (print) | ISBN 9781292139937
(pdf) | ISBN 9781292139944 (epub)
Subjects: LCSH: Chemistry--Study and teaching.
Classification: LCC QD33.2 .P73 2017 | DDC 542--dc23
LC record available at https://lccn.loc.gov/2017002095
10 9 8 7 6 5 4 3 2 1
21 20 19 18 17
Print edition typeset in 10/12pt Times LT Pro by Spi Global

Printed and bound in Malaysia
NOTE THAT ANY PAGE CROSS REFERENCES REFER TO THE PRINT EDITION

Contents
List of boxes viii

Preface to the third edition xi
Guided tour xii
For the student xiv
Acknowledgements xv
List of abbreviations xvi
The investigative approach 1

1. Essentials of practical work 3
2. Health and safety 6
3. Making measurements 36
4. SI units and their use 40
5. Scientific method and design of experiments 46
6. Making notes of practical work 52
7. Project work 56
Fundamental laboratory techniques 61

8. Working with liquids 63
9. Basic laboratory procedures I 70
10. Basic laboratory procedures II 82
11. Principles of solution chemistry 102
12. pH and buffer solutions 113
Laboratory techniques 121

13. Melting points 123
14. Recrystallisation 128
15. Solvent extraction 139
16. Distillation 145
17. Reflux 155
18. Evaporation 161
19. Inert atmosphere methods 166
20. Combinatorial chemistry 173
Classical techniques 177

21. Qualitative techniques for inorganic analysis 179
22. Gravimetry 184
23. Molecular formulae 187
24. Procedures in volumetric analysis 190
25. Acid–base titrations 199
26. Complexometric titrations 203
27. Redox titrations 208
28. Precipitation titrations 211

Contents
Instrumental techniques 215

29. Basic spectroscopy 217
30. Atomic spectroscopy 226
31. X-ray fluorescence spectroscopy 243
32. Chromatography 250
33. Electrophoresis 287
34. Electroanalytical techniques 302
35. Using radioisotopes 309
36. Infrared and Raman spectroscopy 318
37. Nuclear magnetic resonance spectrometry 332
38. Mass spectrometry 353
39. X-ray diffraction 362
40. Thermal analysis 370
Information technology and library resources 373

41. Finding and citing published information 375
42. Evaluating information 382
43. Using online resources 389
44. Internet resources 399
45. Using spreadsheets 421
46. Using word processors, databases and other packages 428
Analysis and presentation of data 435

47. Fundamental principles of quantitative chemical analysis 437
48. Calibration and quantitative analysis 441
49. Using graphs 447
50. Presenting data in tables 459
51. Hints for solving numerical problems 463
52. Manipulating and transforming raw data 472
53. Descriptive statistics 476
54. Choosing and using statistical tests 487
55. Drawing chemical structures 501
56. Chemometrics 507
57. Computational chemistry 514
Study and examination skills 521

58. The importance of transferable skills 523
59. Managing your time 529
60. Working with others 534
61. Taking notes from lectures and texts 538
62. Learning effectively 544
63. Revision strategies 551
64. Assignments and exams 556
65. Preparing your curriculum vitae 566
vi

Contents
Communicating information 573

66. Organising a poster display 575
67. Giving a spoken presentation 581
68. General aspects of scientific writing 587
69. Writing essays 594
70. Reporting practical and project work 597
71. Writing literature surveys and reviews 606
Answers to study exercises 609

Index 627
vii

List of boxes
2.1 How to perform a risk matrix analysis 8

4.1 Conversion factors between some redundant units and the SI 42
4.2 How to interconvert SI units 44
5.1 Checklist for designing and performing an experiment 49
7.1 How to write a project proposal 57
8.1 Using a pipettor to deliver accurate, reproducible volumes of liquid 66
9.1 How to make up an aqueous solution of known concentration from
a solid chemical 72
9.2 How to make up an aqueous solution of known concentration from
a solid chemical for use in quantitative analysis 74
9.3 How to make up a linear dilution series for use in quantitative analysis 77
9.4 How to weigh out a sample of a solid for use in quantitative analysis 80
10.1 How to flute a filter paper for gravity filtration 84
10.2 Isolation of a solid by suction filtration 86
10.3 How to dry a solution over magnesium sulphate 98
11.1 Useful procedures for calculations involving molar concentrations 103
11.2 How to convert ppm into mass of chemical required 104
11.3 An example of complex formation 108
11.4 The use of oxidation numbers to identify redox systems 110
11.5 How to balance redox equations from partial ionic equations using the
ion–electron method 110
12.1 Using a glass pH electrode and meter to measure the pH of a solution 115
14.1 How to carry out a solvent selection for recrystallisation of an unknown compound 131
14.2 How to carry out a mixed-solvent selection for recrystallisation
of an unknown compound 132
14.3 How to carry out a single-solvent recrystallisation 133
14.4 How to carry out a mixed-solvent recrystallisation 135
15.1 How to separate a carboxylic acid and a hydrocarbon using solvent extraction 142
15.2 How to set up a Soxhlet extraction system 143
16.1 How to assemble the apparatus for a simple distillation 147
16.2 How to carry out a simple distillation 147
16.3 How to carry out a fractional distillation 148
16.4 How to carry out a reduced-pressure distillation using a water pump 150
16.5 How to carry out a steam distillation 153
17. 1 How to set up a simple reflux apparatus 156
17. 2 How to set up the apparatus for reflux with mechanical stirring 157
18.1 How to use a rotary film evaporator 162
19.1 How to transfer an air-sensitive reagent using a syringe 170
21.1 How to use a low-speed bench centrifuge 181
22.1 How to carry out gravimetric analysis 184
23.1 How to calculate a molecular formula from microanalytical data 188
24.1 Types of calculations used in volumetric analysis – titrations 191
24.2 How to fill a pipette 195
24.3 How to carry out a titration 195
29.1 How to use a UV/visible spectrophotometer 220
29.2 How to use a flame photometer 223
30.1 How to prepare a 1000 mg mL-1 stock solution of a metal ion from a metal salt 226
30.2 How to prepare a set of five calibration solutions in the concentration range
0910 mg mL-1 (mg L-1) 227
30.3 How to analyse a sample using the method of standard additions in FAAS 229
30.4 Sample size and certified reference materials 229
viii

List of boxes
30.5 Analysis of a sample: dilution factor 230

30.6 How to operate a flame atomic absorption spectrometer 233
30.7 How to acid-digest a sample using a hot plate 240
31.1 How to avoid problems with liquid samples in XRF 247
31.2 How to prepare a loose powder sample for XRF analysis 249
32.1 How to prepare a set of five calibration solutions in the concentration range
0910 mg mL-1 (mg L-1) 257
32.2 How to make micropipettes for TLC 261
32.3 How to run a TLC 262
32.4 How to prepare and run a flash column 264
32.5 How to use a Soxtec extractor 273
32.6 Procedure for Shake Flask extraction 274
32.7 Procedure for ultrasonic extraction 275
32.8 How to operate a typical supercritical fluid extraction system 275
32.9 How to operate a typical pressurised microwave-assisted extraction (MAE) system 276
32.10 How to operate a typical pressurised fluid extraction (PFE) system 277
32.11 How to use a low-speed bench centrifuge 277
32.12 How to pre-concentrate a sample using a reversed phase C18 solid phase
extraction (SPE) cartridge 280
32.13 How to concentrate a sample of an organic compound in an aqueous sample
using direct solid phase microextraction (SPME) 281
32.14 How to concentrate a sample of an organic compound in an aqueous sample
using headspace solid phase microextraction (SPME) 281
33.1 How to carry out agarose gel electrophoresis of DNA 290
33.2 How to carry out SDS-PAGE for protein separation 294
35.1 How to determine the specific activity of an experimental solution 312
35.2 Tips for preparing samples for liquid scintillation counting 314
36.1 How to run an infrared spectrum of a liquid, solid film, mull or KBr disk 321
36.2 How to prepare liquid and solid films and mulls 323
36.3 How to prepare a KBr disk 324
36.4 How to interpret an IR spectrum 327
37.1 How to prepare a sample for NMR spectroscopy 335
37.2 How to interpret a 1H9 1H@COSY spectrum 349
38.1 How to identify the number of bromine or chlorine atoms in a molecule
from the molecular ion 355
38.2 Idealised fragmentation processes for the molecular ion (M +) 356
39.1 Press and Pull method for mounting of specimen 364
39.2 Back-loading method for mounting of specimen 364
39.3 Side-loading method for mounting of specimen 365
40.1 How to interpret a thermal analysis trace 371
42.1 How to avoid plagiarism and copyright infringement 383
43.1 Important guidelines for using PCs and networks 390
43.2 Getting to grips with e-learning 391
43.3 Useful tips for using search engines 394
43.4 Getting the most from Google searches 395
43.5 How to evaluate information on the Web 396
48.1 The stages involved in preparing and using a calibration curve 442
48.2 How to use a spreadsheet (a Microsoft Excel ) to produce a linear regression plot 444
49.1 Checklist for the stages in drawing a graph 450
49.2 How to create and amend graphs within a spreadsheet (Microsoft Excel )
for use in coursework reports and dissertations 450
49.3 How graphs can misrepresent and mislead 456
50.1 Checklist for preparing a table 460
50.2 How to use a word processor (Microsoft Word) or a spreadsheet (Microsoft Excel)
to create a table for use in coursework reports and dissertations 461
ix

List of boxes
5 1.1 Example of using the algebraic rules of Table 51.2 465

53.1 Descriptive statistics for a sample of data – an example 478
53.2 Three examples where simple arithmetic means are inappropriate 479
53.3 How to use a spreadsheet (Microsoft Excel) to calculate descriptive statistics 484
54.1 How to carry out a t-test 492
54.2 Worked example of a t-test 493
54.3 Using a spreadsheet (Microsoft Excel) to calculate hypothesis-testing statistics 497
56.1 Example of a two-level factorial design 509
56.2 Example of principal component analysis 512
58.1 How to carry out a personal skills audit 526
59.1 Tips for effective planning and working 532
61.1 The SQ3R technique for skimming texts 542
62.1 How to diagnose your learning preferences using the VAK scheme 546
62.2 How to accommodate different lecturers, teaching styles 548
63.1 How to use past exam papers in your revision 553
63.2 How to prepare and use a revision timetable 553
63.3 How to revise actively 554
64.1 Problem-based learning (PBL) 557
64.2 Writing under exam conditions 559
64.3 Reasons for poor exam answers to essay-style questions 560
64.4 Strategies for combating the symptoms of exam anxiety 564
65.1 The structure and components of a typical CV and covering letter 568
66.1 How to create a poster using Microsoft PowerPoint 579
67.1 Tips on preparing and using Microsoft PowerPoint slides in a spoken presentation 582
67.2 Hints on spoken presentations 585
68.1 How to achieve a clear, readable style 590
68.2 Using appropriate writing styles for different purposes (with examples) 591
68.3 How to improve your writing ability by consulting a personal reference library 592
70.1 The structure of reports of experimental work using the ‘IERaD’ structure 598
70.2 Writing experimental procedures 600
70.3 How to write up your research project (dissertation) or thesis 602
70.4 Steps in producing a scientific paper 604
71.1 How to analyse a topic using the SPSER approach 607

Preface to the third edition
‘Chemistry can be defined as the science that studies practical skills. Specifically, ‘to develop in students a range
systematically the composition properties, and reactivity of practical skills so that they can understand and assess
of matter at the atomic and molecular level. Since matter risks and work safely and competently in the laboratory’
is everything that can be touched, made visible, smelt or [for undergraduate students] and ‘to provide students with
tasted, it follows that the scope of chemistry as a subject the ability to plan and carry out experiments independently
is very broad’. and assess the significance of outcomes’ [for postgraduate
students].
To students who buy this book, we hope you will find
QAA for HE Subject Benchmark it useful in the laboratory during your practical classes and
Statement for Chemistry (2014). in your project work – this is not a book to be left on the
Practical skills form the cornerstone of chemistry. Howev- bookshelf.
er, the diversity of skills required in the laboratory means We would like to take this opportunity to thank our wives
that a student’s experience may be limited. While some and families for their continued support, and to recognise
techniques do require specific skills, many of them are the following colleagues and friends who have provided as-
transferable generic skills that are required throughout the sistance, comment and food for thought at various points
subject area. during the production of all editions: Gary Askwith, Dave
The time constraints of the modern curriculum often Bannister, Jon Bookham, Samantha Bowerbank, Susan
preclude or minimise laboratory time. It is the aim of this Carlile, Michelle Carlin, Jim Creighton, Sarah
Cresswell,
book to provide general guidance for use in and out of prac- Martin Davies, Mike Deary, Les Dix, M arcus Durrant,
tical sessions and also to cover a range of techniques from Jackie Eager, Gordon Forrest, Derek Holmes, Ed Ludkin,
the basic to the more advanced. Dave Osborne, Justin Perry, Lee Rounds, Jane Shaw, Tony
In creating the third edition of Practical Skills in Chem- Simpson, Dave Wealleans and Ian Winship. We would also
istry, we have maintained the approach of the previous like to thank the staff of Pearson Education for the friend-
editions, with the aim of providing support to students tak- ly support over the years, and would wish to acknowledge
ing chemistry based courses in a concise and user friendly Richelle Zakrewski, Rufus Cornow, Pat Bond, Owen Knight,
manner. Key points, definitions, illustrations, ‘how to’ box- Simon Lake, Alex Seabrook and Pauline Gillett.
es, checklists, worked examples, tips and hints are includ- As with previous editions, we would be grateful to
ed where appropriate. However, we have also used this hear of any errors you might notice, so that these can be
opportunity of the new edition to restructure the layout, put right at the earliest opportunity.
to literally start at the beginning of the laboratory process
and progress to the end, with the dissemination of results.
In updating and thoroughly revising the book to in-
clude a ‘taste’ of the latest developments in methodology, JOHN R. DEAN (john.dean@northumbria.ac.uk)
we have considered carefully the Quality Assurance Agen- ALAN M. JONES (alanjones559@btinternet.com)
cy UK Subject Benchmarking statements for Chemistry, re-
viewed and updated in 2014, and have attempted to cover DAVE HOLMES (david.holmes@northumbria.ac.uk)
all of the generic skills, along with the practical aspects ROB REED (r.reed@cqu.edu.au)
of the subject specific topics in chemistry. We have been
ALLAN M. JONES (a.m.jones@dundee.ac.uk)
mindful of two of the QAA’s aims for chemistry degree
(under- and post-graduate) programmes in the context of JONATHON WEYERS (j.d.b.weyers@dundee.ac.uk)

xi

Guided tour
43 Using online resources
Information and communication technology (ICT) is vital in the modern aca-

Definitions demic world and ‘IT literacy’ is a core skill for all bioscientists. This involves
Definitions of key terms and

a wide range of computer-based skills, including:
Browser – a program to display web
pages and other Internet resources. ●● Accessing web pages using a ‘browser’ such as Internet Explorer, Firefox
Safari or Chrome.
concepts are highlighted in FAQ – Frequently Asked Question; a file
or web page giving information on com- ●● Searching the web for useful information and resources using a search
engine such as Google, or a meta-search engine such as Dogpile.
mon queries, sometimes used as a file
the text margin. extension (.faq).
FTP – File Transfer Protocol; a mecha-
●● Finding what you need using online databases, such as library catalogues
or complex websites, such as your university’s homepage.
nism for downloading files. ●● Downloading, storing and manipulating files.
URL – Uniform Resource Locator; the ●● Communicating via the Internet.
‘address’ for web resources.
●● Using e-learning facilities effectively.
●● Working with ‘Office’-type programs and other software (dealt with in
detail in Chapters 45 and 46).
Academic use of ICT resources – a
range of in appropriate activities will be You will probably receive an introduction to your university’s networked
identified in your university’s rules for IT systems and you will be required to follow rules and regulations that are
use of ICT systems. They may include: important for the operation of these systems. Whatever your level of experience
hacking, spamming, using another per- with PCs and the Internet, you should also follow the basic guidelines shown
son’s account, and copyright infringe- in Box 43.1. Reminding yourself of these from time to time will reduce your
ment, as well as broader aspects of chances of losing data.
behaviour covered by a code of conduct
or student charter The Internet as a global resource
The Internet is a complex network of computer networks; it is loosely organised
and no one group organises it or owns it. Instead, many private organisations,
universities and government organisations fund and operate discrete parts of it.
The web is the most popular application of the Internet. It allows easy
Tips and Hints provide use- links to information and files which may be located on networked computers
across the world. The web enables you to access millions of ‘homepages’ or
ful hints and practical advice,

‘websites’ – the initial point of reference with many individuals, institutions
and companies. Besides text and images, these sites may contain ‘hypertext
links’, highlighted words or phrases that take you to another Internet location
and are highlighted in the text Understanding the technology – you

do not need to understand the work-
via a single mouse click.
You can gain access to the Internet either through a network at your uni-
versity, at most public libraries, at a commercial ‘Internet cafe’, or from home
margin. ings of the Internet to use it – most of
it is invisible to the user. To ensure you
via a modem connected to a broadband or dial-up internet service provider
(e.g. Virgin Media, BT or Sky).
obtain the right facilities, you may need
to know some jargon, such as terms for
Key PoInT Most material on the Internet has not been subject
Key Points highlight critical
the speed of data transfer (megabits)
and the nature of internet addresses. to peer review or vetting. Information obtained from the web or
Setting up a modem and/or local posted on newsgroups may be inaccurate, biased or spoof; do
features of methodology. wireless network can be complex, but

instructions are usually provided with
not assume that everything you read is true, or even legal.
the hardware. White and Downs (2014) online communication

and Gralla (2006) are useful texts if you
wish to learn more about computing
You will be allocated an email account by your university and should use
and the Internet.
this routinely for communicating with staff and fellow students, rather than
using a personal account. You may be asked to use email to submit work as
M43 Practical Skills in Chemistry 39920.indd 389 10/05/2017 20:22
Basic laboratory procedures I
35 Using radioisotopes Box 9.4 How to weigh out a sample of a solid for use in
quantitative analysis
The isotopes of a particular element have the same number of protons in the 1. Place the clean, dry weighing boat or sample tube on a general-
Examples 126 C, 136 C and 146 C are three of purpose two-decimal-place balance and zero the balance.
nucleus but different numbers of neutrons, giving them the same proton number
the isotopes of carbon. About 98.9% (atomic number) but different nucleon numbers (mass number, i.e. number of 2. Weigh out the calculated amount of chemical within the accuracy of
of naturally occurring carbon is in the
protons + number of neutrons). Isotopes may be stable or radioactive. Radi- the balance.
stable 126 C form. 136 C is also a stable
oactive isotopes (radioisotopes) disintegrate spontaneously at random to yield
isotope but it only occurs at 1.1% natu- 3. Check the zero reading on the analytical balance by pressing the bar/
ral abundance. Trace amounts of radio- radiation and a decay product. button with the balance doors closed.
active 146 C are found naturally; this is a
4. Relock the balance pan by pressing the bar/button.
negatron-emitting radioisotope. Radioactive decay
5. Carefully transfer the weighing boat or sample tube to the balance
There are three forms of radioactivity (Table 35.1) arising from three main
pan of the analytical balance (for very accurate work use tweezers
types of nuclear decay: or fine tongs since the sweat from your fingers will contribute to the
●● Alpha decay. This involves the loss of a particle equivalent to a helium weight recorded) and close the balance door.
Examples 226Ra decays to 222Rn by nucleus. Alpha (a) particles, being relatively large and positively charged,
loss of an alpha particle, as follows:
6. Release the balance pan by pressing the bar/button, allow the balance
do not penetrate far in living tissue, but they do cause ionisation damage and to stabilise and record the weight of the chemical and container. If the
226
88 Ra S 222 4
86 Rn + 2He
2+ this makes them generally unsuitable for tracer studies. last decimal place ‘cycles’ between two or three numbers, determine
the mid-point of the ‘cycle’ and record this value as the weight.
14
C shows beta decay, as follows: ●● Beta decay. This involves the loss or gain of an electron or its positive
14 S 147 N + b-
counterpart, the positron. There are three subtypes: 7. Lock the balance pan by pressing the bar/button, remove the sample
6C
container and transfer the solid to your volumetric flask, beaker or
22 (a) Negatron (b- ) emission: loss of an electron from the nucleus when
Na decays by positron emission, as conical flask by pouring, but do not wet the weighing boat or sample
follows: a neutron transforms into a proton. This is the most important form tube with solvent.
of decay for radioactive tracers used in chemistry. Negatron-emitting
22 S 22 +
11Na 10Ne + b isotopes of importance include 3H, 14C, 32P and 35S. 8. Replace the weighing container on the analytical balance pan, close
55 the balance door and weigh the container. Again decide on the mid-
Fe decays by electron capture and (b) Positron (b+ ) emission: loss of a positron when a proton transforms into point weight if the last decimal place ‘cycles’ and record this value
the production of an X-ray, as follows: a neutron. This only occurs when sufficient energy is available from as the weight of the ‘empty’ weighing container.
55
26Fe S 55
25Mn + X the transition and may involve the production of gamma rays when the
positron is later annihilated by collision with an electron. 9. Lock the balance pan by pressing the bar/button and remove the
The decay of 22Na by positron emis- weighing container from the balance.
sion (b+) leads to the production of a (c) Electron capture (EC): when a proton ‘captures’ an electron and trans- Fig. 9.5 Transferring a solid using glazed
g ray when the positron is annihilated paper. 10. Subtract the weight of the ‘empty’ weighing container from that of
forms into a neutron. This may involve the production of X-rays as
on collision with an electron. the weighing container plus sample and you now know the mass
electrons ‘shuffle’ about in the atom (as with 125I) and it frequently
of chemical, to an accuracy of four decimal places, which has been
involves electron emission.
transferred into your volumetric flask, beaker or conical flask.
●● Gamma emission. Internal transition involves the emission of electromag-
netic radiation in the form of gamma (g) rays from a nucleus in a metastable
state and always follows initial alpha or beta decay. Emission of gamma amounts of solid are to be transferred, it is advisable to use a wide-necked filter
folded glazed paper
radiation leads to no further change in atomic number or mass. funnel called a ‘powder funnel’.
solid In many preparative experiments, which are carried out on a small scale
Note from the above that more than one type of radiation may be emitted (involving 1 g to 10 g of solids), the most useful weighing container is spe-
when a radioisotope decays. The main radioisotopes used in chemistry and rolled glazed paper
cial glazed paper, provided that the chemicals do not react with the paper.
their properties are listed in Table 35.2. A creased square of glazed paper is ‘tared’ on the balance pan and the solid
weighed out directly onto it. The chemical can then be allowed to flow down
Table 35.1 Types of radioactivity and their properties. the crease into the vessel (Fig. 9.5). Furthermore, when attempting to transfer
small amounts of solid in vessels with narrow-bore ground-glass joints (see
Range of maximum Penetration range Suitable shielding flask p. 98) it is important not to allow the solid to contact the joint, because the joint
Radiation energies (MeV*) in air (m) material will not seal correctly. Use a filter funnel or roll a piece of glazed paper into
Alpha (a) 4–8 0.025–0.080 Unnecessary a funnel, insert the stem of the paper funnel to below the joint and then run in
the solid from the creased weighing paper (Fig. 9.6). Paper used in this manner
Beta (b) 0.01–3 0.150–16 Plastic (e.g. Perspex)
Fig. 9.6 Transferring a solid to a narrow-necked is much cheaper than proprietary weighing dishes and is a useful method of
Gamma (g) 0.03–3 1.3–13† Lead flask. recycling out-of-date manufacturers’ catalogues!
*Note that 1 MeV = 1.6 * 10 -13 J.
† Distance at which radiation intensity is reduced to half. 80 Fundamental laboratory techniques
M35 Practical Skills in Chemistry 39920.indd 309 10/05/2017 19:58 Worked examples and ‘How to’ boxes
Examples are included in the margin to illustrate important set out the essential procedures in a
points without interrupting the flow of the main text. step-by-step manner.
xii

Guided tour
Chromatography
100% dimethylsiloxane:
the least polar bonded phase. Used for boiling point
separations (solvents, petroleum products, etc.).
Typical names: DB–1, HP–1, Rtx–1
CH3
Si O
CH3
Figures are used to illustrate (a) (b)
95% dimethylsiloxane–5% diphenylypolysiloxane:
key points, techniques and a non-polar phase. Used for separation of environmental
samples, e.g. polycyclic aromatic hydrocarbons.
equipment. CH3
Si O Si O
CH3
(c) (d) 5% 95%
Fig. 32.11 Sample injection in GC. (a) Fill the syringe, (b) wipe clean the Fig. 32.13 Common stationary phases for
Chromatography
outside of the syringe needle, (c) place the syringe needle into the injector capillary GC.
and (d) depress the plunger on the syringe to inject the sample.
Polyimide syringe
coating
Fused-silica PTFE block
Stationary
phase
syringe
Fig. 32.12 Schematic diagram of a GC column.

to purge
valve ball valve
septum
Box 32.1 How to prepare a set of five calibration solutions in the concentration range
0910 mg mL −1 (mg L -1)
syringe needle
glass liner
Assuming that we are starting with a 1000 mg mL-1 [32.9]
column end
stock solution of a particular organic compound (e.g.
carrier in the 10.00 mL
so 20 mg 2-chlorophenol was placed
2-chlorophenol) you will need the following: 6 * 10.00 mL gas inlet
grade A volumetric
column flasks and a syringe (09100.00 mL). volumetric flask. So,
compressed
1. ensure that all the glassware is clean (see p. 68). 20 mg air
= 2 mg mL-12 @chlorophenol
for cooling [32.10]
10 mL
2. Add ?9 mL of organic solvent (e.g. dichloromethane) inside the oven
carrier to split valve
to a 10.00 mL with dichloromethane.
gas inlet
You now have a 2 mg mL-1 calibration solution of
2-chlorophenol.
3. Quantitatively transfer 20.00 mL of the stock solu-
tion into the 10.00 mL volumetric flask. Inject the 5. Similarly transfer 0,syringe
40.00,needle
60.00, 80.00 and 100.00 mL
solution from the syringe below the surface of the volumes into separate volumetric flasks and dilute
dichloromethane. Then, dilute to 10.00 mL with to 10.00 mL with dichloromethane and label as 0,
(a) (b)
dichloromethane. 4, 6, 8 and 10 mg mL-1 2-chlorophenol calibration
Fig. 32.10 Sample introduction in GC: (a) split/splitless injector; (b) solutions.
on-column injector.
4. What is the concentration of this new solution?
Remember that we started with an initial 1000 mg mL-1 6. take the 0, 2, 4, 6, 8 and 10 mg mL-1 2-chlorophe-
2-chlorophenol stock solution. nol calibration solutions to the chromatograph for
1000 mg analysis.
* 20 * 10-3 mL = 20 mg 2@chlorophenol
Fig. 32.12: at normal operating temperatures. This behaves in a similar man-
mL compounds by GC in the
Analysing
ner to a liquid film, but is far more robust. Common stationary phases for GC
split mode – make sure no hazardous
are shown in Fig. 32.13. The mobile phase (‘carrier gas’) is usually nitrogen
materials enter the laboratory atmos- Instrumental techniques
or helium. Selective separation is achieved as a result of the differential parti- 257
phere through the split vent. A charcoal
tioning of individual compounds between the carrier gas and silicone polymer
split-vent trap may be required to elim-
phases. The separation of most organic molecules is influenced by the temper-
inate potential hazards.
ature of the column, which may be constant during the analysis (‘isothermal’ –
usually 50–250 °C) or, more commonly, may increase in a pre-programmed
M32 Practical Skills in Chemistry 39920.indd 257 manner (e.g. from 50 °C to 250 °C at 10 °C per min). 10/05/2017 19:49
Selecting an appropriate column for capillary GC is a difficult task and

SI units and their use one which is usually left to the technician. However, it is important to be Precipitation titrations
aware of some general issues and what influence they can have on the sep-
aration. The column internal diameter can affect both resolution and speed
of analysis. Smaller internal diameters columns (0.25 mm i.d.) can provide
Table 28.2 Selected applications of precip- 3. Fajans titration, which involves the adsorption of a coloured indicator onto
good resolution of early eluting peaks (Fig. 32.14(a)). However, the problem
itation titrations thethat
precipitate at the end-point, resulting in a colour change. During this
Box 4.2 How to interconvert SI units is that the analysis times of the eluting components may be longer and the
linear dynamic Analyte
range may beComments restricted. In contrast, larger internaladsorption
diameter process a change occurs in the indicator resulting in a change
Example: You are required to calculate the molecular 2. Look at the units and decide which are common: columns (0.53 mm i.d.) provide less resolution for early eluting compounds of colour. The indicators used for this are often anionic dyes, e.g. fluores-
weight of a polymer by measurements of its osmotic since the gas constant is expressed in joules, you (Fig. 32.14b), butCl - ,this
Br - is reflected
Mohr in
method: 2CrO4 used
shorterAganalysis cein or lin-
times and a greater eosin. The most common indicator for AgCl is dichlorofluorescein
pressure in solution. At infinite dilution, measured should convert the osmotic pressure term into joules. ear dynamic range. This type as of end-point
column may provide sufficient resolution(Fig. 28.2)
for (this is greenish yellow in solution but changes colour to pink
graphically from your experiments, the equation below The derived unit for pressure is N m -2 and, since the Br - , I - , AsO
the analysis of complex -
mixtures. Fig.
Volhard
4 precipitate the effects when
32.14 illustrates
method: it is adsorbed on AgCl).
of column
applies: derived units for N are J m -1, the full derived unit of removal is unnecessary
internal diameter. Selected examples of precipitation titrations are shown in Table 28.2.
pressure is (J m -1) * m -2 = J m -3.
Π RT Cl - , CN- , Volhard method: precipitate
= CO32- removal is required
c Mr 3. Substitute the units into the equation for M:
256 Instrumental techniques
RTc JK -1
mol -1
* K * kg m -3 Cl - , Br - , I - , Fajans method: titration with
where Π = osmotic pressure at infinite dilution (Pa),
Mr = = = kg mol-1 SCN- Ag+ . Detection with fluorescein,
R = gas constant (J K -1mol-1), T = temperature (K), Π J m -3 dichlorofluorescein and eosin
c = concentration of solution (kg m -3) and Mr = molecular
weight. 4. Substitute the appropriate numerical values into the F- Titration with Th(NO3)4 to
equation for Mr: you know that the units of the cal- produce ThF4. End-point
1. Re-arrange the equation for Mr: M32 Practical Skills in Chemistry 39920.indd
culation will be correct since the molecular weight is
256 detection with alizarin red S 10/05/2017 19:49
the weight of 1 mole of polymer, expressed in kg. Adapted from: Quantitative Chemical Analysis,
RTc
Mr = 4th edn, D.C. Harris, W.H. Freeman, New York
Π
(1995), p. 176.
Temperature
Definitions
The SI unit is the kelvin, K. The degree Celsius scale has units of the same Sources for further study
STP - Standard Temperature and magnitude, °C, but starts at 273.15 K, the melting point of ice at STP. Temper-
Christian, G.D., Dasgupta, P.K. and Schug, K.A. (2014) Chemical Analysis, 6th edn. Prentice Hall, Harlow,
Pressure = 293.15 K and 101325Pa (or ature is similar to time in that the Celsius scale is in widespread use, but note
Analytical Chemistry, 7th edn. John Wiley & Sons Ltd, Essex.
101.325 kPa or 0.101 325 M Pa). that conversions to K may be required for calculations. Note also that you must
Chichester. McPherson, P. (2014) Practical Volumetric Analysis. RSC,
not use the degree sign (°) with K and that this symbol must be in upper case
to avoid confusion with k for kilo; however, you should retain the degree sign Harris, D.C. (2010) Quantitative Chemical Analysis, 8th Cambridge.
with °C to avoid confusion with the coulomb, C. edn. W.H. Freeman Co., New York. Rubinson, J.F. and Rubinson, K.A. (2003) Contemporary
Jander, G., Jahr, K.-F., Schulze, G. and Simon, J. (2009) Volu- Chemical Analysis. Pearson, Harlow, Essex.
Interconversion of SI units metric Analysis, 17th edn. Walter de Gruyter Inc., Germany. Skoog, D.A., West, D.M., Holler, F.J. and Crouch, S.R.
You will find that the use of SI units simplifies mathematical manipulations Mendham, J., Denney, R.C., Barnes, J.D. and Thomas, (2014) Fundamentals of Analytical Chemistry, 9th edn.
and ensures that you obtain the correct units for the parameter being calculated. M.J.K. (2000) Vogel ’s Textbook of Quantitative Brooks Cole, Belmont, CA.
Remember that you must convert all units into the appropriate SI units, e.g.
masses must be expressed as kg, volumes as m3 and concentrations as kg m -3
or mol m -3, etc., and that you may need to use alternatives in derived units
(Table 4.2). The application of these principles is shown in Box 4.2.
Study exercise
Sources for further study 28.1 You are a major manufacturer of fireworks and ‘potassium nitrate’ (4.0124 g) was dissolved
Anon. (2000) The NIST Reference on Constants, Units and Anon. (2014). Measurement units: the SI. Available: you suspect that your bulk supplier of potas- in water and made up to 250.00 mL. This solu-
Uncertainty. Available: http://physics.nist.gov/cuu/units/ www.bipm.org/en/measurement-units/ sium nitrate, a white crystalline solid, has tion (25.00 mL) required silver nitrate solution
index.html Last accessed 05/01/16. been adulterating the potassium nitrate with (10.3 mL; 0.1 M) for equivalence using dichlor-
Last accessed 05/01/16. [Online information includes the SI brochure: The salt to increase his profits. Your analysis of the ofluorescein as indicator. Calculate the % (w/w)
Anon. Measurement units. Available: www.npl.co.uk/ International System of Units (SI), 9th edn (draft).] ‘potassium nitrate’ gave the following results: salt in the ‘potassium nitrate’.
reference/measurement-units/ Blackman, A. and Gahan, L. (2014) Aylward and Findlay’s
Last accessed 05/01/16. SI Chemical Data, 7th edn. John Wiley & Sons Ltd,
Chichester.
44 The investigative approach Classical techniques 213
M04 Practical Skills in Chemistry 39920.indd 44 10/05/2017 15:49 M28 Practical Skills in Chemistry 39920.indd 213 11/05/2017 09:12
Sources for further study – every chapter is Study exercises are included in every chapter to
supported by a section giving printed and electronic reinforce learning with problems and practical exer-
sources for further study. cises.
xiii

For the student
This book aims to provide guidance and support over the spectroscopy), to separation techniques (chromatography
broad range of undergraduate courses, as well as some and electrophoresis), to electrochemistry, use of radioiso-
postgraduate courses, including laboratory classes, project topes and structural techniques (infrared spectroscopy, nu-
work, lectures, tutorials, seminars and examinations, as clear magnetic resonance spectrometry, mass spectrome-
outlined below: try, X-ray diffraction and thermal analysis).
Chapters 1–7 (The investigative approach) Chapters 41–57 (IT, internet and data
Introduce the initial key aspects of all laboratory work. analysis)
Specifically, the essentials of all practical work, health and Cover all aspects of data, from finding useful and relevant
safety aspects (Risk Assessment and COSHH), making information to solving a problem to useful references on
measurements, SI Units and their use, scientific method ‘how to’ perform statistical tests.
and design of experiments, making notes of practical exer-
cises and project work.
Chapters 58–65 (Study and examination
skills)
Chapters 8–12 (Fundamental laboratory
techniques) Focus on the specific skills that will allow you to work ef-
fectively to achieve optimum success during your course
Cover all aspects of laboratory procedures including work-
and beyond.
ing with liquids, solution chemistry and pH and buffer solu-
tions.
Chapters 66–71 (Communicating
Chapters 13–28 (Laboratory techniques) information)
Introduce all the basic laboratory techniques for use Are key to success in chemistry; these chapters provide
in chemistry. Their contents range from basic tech- the essential components that you need to consolidate or
niques used in synthetic chemistry (e.g. melting point, improve upon to succeed.
recrystallisation, solvent extraction, distillation, reflux and
evaporation) through to more advanced areas (e.g. inert Study exercises
atmosphere techniques and combinatorial chemistry). In Provide a valuable resource to allow you to practice and
addition, classical techniques for qualitative inorganic analysis revise key aspects of selected chapters. Answers are
are covered as well as quantitative approaches (including provided at the back of the book. For numerical exercis-
gravimetry, molecular formulae and titrimetry techniques). es, the working out is also provided, as well as the final
answer. In some cases, the answer is in the form of tips
Chapters 29–40 (Instrumental techniques) to allow you to investigate further or provide the direc-
Cover essential relevant analytical instrumental techniques tion for a suitable answer.
from the analysis of molecules (basic spectroscopy), ele- We hope that you find this book a useful resource
mental analysis (atomic spectroscopy, X-ray fluorescence throughout your chosen course, and beyond.
xiv

Acknowledgements
We are grateful to the following for permission to repro- Chemistry; Screenshots 44.4, 44.5 from www.rsc.org/
duce copyright material: merck-index, reproduced with permission from the Royal
Society of Chemistry; Screenshots 44.6, 44.7 from www.
EPSRC funded National Chemical Database Service hosted
chemspider.com, reproduced with permission from the
by the Royal Society of Chemistry; MassBank: a public repos-
Royal Society of Chemistry; Screenshots 44.10, 44.11
itory for sharing mass spectral data for life sciences; North-
from http://www.massbank.jp/, reproduced with permis-
umbria University for the Risk Assessment, COSHH (short)
sion of MassBank Project.
and COSHH (extended) forms; and, Sigma Aldrich Ltd. for
Hazards Statements, Precautionary Statements, Pictograms
– hazard codes and MSDS for phenol (as an example). Tables
While every effort has been made to trace the own- Table 2.1 from http://www.sigmaaldrich.com/help-
ers of copyright material, in a few cases this has proved welcome/hazard-and-precautionary-statements.html,
impossible and we take this opportunity to offer our apol- reproduced with permission from Merck; Table 2.4 from P
ogies to any copyright holders whose rights we may have Statements, http://www.sigmaaldrich.com/help-welcome/
unwittingly infringed. hazard-and-precautionary-statements.html, reproduced
with permission from Merck; Table 44.4 from http://
Figures usefulchem.wikispaces.com/EXP284, McBride, M.J. and
Figure 2.4 from P Statements, http://www.sigmaaldrich. Bradley, J.C., work was completed using Open Notebook
com/help-welcome/hazard-and-precautionary-statements. Science under the supervision of the late Dr. Jean-Claude
html, reproduced with permission from Merck; Figure 2.5 Bradley.
from www.sigmaaldrich.com, reproduced with permis-
sion from Merck; Figure 33.11 from Protein concentration
Picture Credits
by precipitation with pyrogallol red prior to electropho-
resis, Electrophoresis (Marshall, T., Abbott, N.J., Fox, P., The publisher would like to thank the following for their
and Williams, K.M. 1995), Courtesy of Marshall, T. and kind permission to reproduce their photographs:
Williams, K.M., International Electrophoresis Society,
(Key: b-bottom; c-centre; l-left; r-right; t-top)
reproduced with permission of John Wiley & Sons Inc. via
Copyright Clearance Center; Figures 33.13, 33.14 from 123RF.com: Vitaliy Malievsky 240b; John Dean: 175/20.2,
http://www.beckmancoulter.com/products/splashpage/ 240tl/30.21, 247/31.8, 247/31.9, 248/31.13, 248br/31.14,
chiral38/default.asp/, Copyright © 1909–2002, Beckman 272/32.24, 316/35.5; Science Photo Library Ltd: Klaus
Coulter, Inc. Guldbrandsen 274t; Shutterstock.com: Strelch 240bl
/30.22
Screenshots
Screenshots 44.1, 44.2, 44.3 from http://cds.rsc.org, All other images © Pearson Education
reproduced with permission from the Royal Society of

xv

List of abbreviations
A absorbance EOF electroosmotic flow

AAS atomic absorption spectroscopy ESI electrospray ionisation
AC affinity chromatography
ACN acetonitrile F Faraday constant
ACS American Chemical Society FAAS flame atomic absorption spectroscopy
AES atomic emission spectroscopy FID flame ionisation detector
ANOVA analysis of variance FT Fourier transform
AO atomic orbital FT–IR Fourier transform–infrared spectroscopy
APCI atmospheric pressure chemical ionisation GC gas chromatography
Ar relative atomic mass GC–MS gas chromatography–mass spectrometry
ASE accelerated solvent extraction GFC gel filtration chromatography
ATP adenosine triphosphate GPC gel permeation chromatography
ATR Attentuated Total Reflection
h Planck constant
b.pt. boiling point HASAW hazards at work
CCD central composite design HCB hexachloro-1,3-butadiene
CCP cubic close packed HCL hollow cathode lamp
CE capillary electrophoresis HCP hexagonal close packed
CEC capillary electrochromatography HEPES N-(2-hydroxyethyl)-N’-piperazine ethane
CGE capillary gel electrophoresis sulphonic acid
CI chemical ionisation HIC hydrophobic interaction chromatography
COSHH control of substances hazardous to health HPLC high performance liquid chromatography
COSY Correlation Spectroscopy HS headspace
CoV coefficient of variation HRMS high resolution mass spectrum
CRM certified reference material HTML hypertext markup language
CW continuous wave ICP inductively coupled plasma
CZE capillary zone electrophoresis ICP–MS inductively coupled plasma–mass
spectrometry
dp decimal point
IEC ion exchange chromatography
DAD diode array detection
IEF isoelectric focusing
DCM dichloromethane
IR infrared (radiation)
DEPT distortionless enhancement by polarisation
transfer ISE ion selective electrode
DNA deoxyribonucleic acid IUPAC International Union of Pure and Applied
Chemistry
dpm disintegrations per minute
DSC differential scanning colorimetry Ka acid dissociation constant
DTA differential thermal analysis kg kilogram
DVB divinylbenzene Kow octanol–water partition coefficient
Ks solubility product
ECD electron capture detector
Kw ion product of water
EDTA ethylenediaminetetraacetic acid
EI electron impact (ionisation) LC–MS liquid chromatography–mass spectrometry
EIE easily ionisable element LGC Laboratory of the Government Chemist
EF empirical formula LOD limit of detection
EMR electromagnetic radiation LOQ limit of quantitation
en ethylenediamine LRMS low resolution mass spectrum
xvi

List of abbreviations
m.pt. melting point RA relative abundance

MAE microwave assisted extraction Rf relative frontal mobility
MDL minimum detectable level RNA ribonucleic acid
MEKC micellar electrokinetic chromatography RP–HPLC reversed phase high performance liquid
MEL maximum exposure limit chromatography
MF molecular formula rpm revolutions per minute
MO molecular orbital RSC Royal Society of Chemistry
Mr relative molecular mass RSD relative standard deviation
MS mass spectrometry SAX strong anion exchange
NH null hypothesis SCOT support coated open tubular (column)
NIST National Institute of Standards and SCX strong cation exchange
Technology SDS sodium dodecyl sulphate
NMR nuclear magnetic resonance SE standard error (of the sample mean)
NP–HPLC normal phase high performance liquid SEM scanning electron microscopy
chromatography SFE supercritical fluid extraction
SI Système Internationale D’Unités
ODS octadecylsilane SPE solid phase extraction
OEL occupational exposure standard SPME solid phase microextraction
PAGE polyacrylamide gel electrophoresis STP standard temperature and pressure
PCA principal component analysis TCA trichloroacetic acid
pdf portable document format TCD thermal conductivity detector
PDMS polydimethylsiloxane TG thermogravimetry
PEEK poly(etheretherketone) TLC thin layer chromatography
PFA perfluoroalkoxyvinylether TMS tetramethylsilane
PFA perfluoroalkoxy fluorocarbon TOF-MS time-of-flight mass spectrometry
PFE pressurised fluid extraction TRIS tris(hydroxymethyl)aminomethane or
pH log10 proton concentration (activity) 2-amino-2-hydroxymethyl-1,3-propanediol
PLOT porous layer open tubular (column)
UKAS United Kingdom Accreditation Services
PMT photomultiplier tube
URL uniform resource locator
ppb parts per billion (109)
USEPA United States Environmental Protection
PPE personal protection equipment Agency
ppm parts per million (106) UV ultraviolet
PTFE polytetrafluoroethylene
WCOT wall-coated open tubular (column)
QA quality assurance www World Wide Web
R universal gas constant z net charge on an ion
xvii

The investigative approach
1. Essentials of practical work 3
2. Health and safety 6
3. Making measurements 36
4. SI units and their use 40
5. Scientific method and design of experiments 46
6. Making notes of practical work 52
7. Project work 56

1 Essentials of practical work
All knowledge and theory in science has originated from practical observation
Developing practical skills – these will
and experimentation: this is equally true for chemical disciplines as diverse as
include:
analysis and synthesis. Laboratory work is an essential part of all chemistry
●● designing experiments
●● observing and measuring
courses and often accounts for a significant proportion of the assessment marks.
●● recording data
The skills and abilities developed in practical classes will continue to be useful
●● analysing and interpreting data throughout your course and beyond, some within science and others in any
●● reporting/presenting. career you choose (see Chapter 58).
Being prepared
Safety Note Mobile phones –
these should never be used in a lab Key Point You will get the most out of laboratory work if you
class, as there is a risk of contamination prepare well. Do not go into a practical session assuming that
from hazardous substances. Always everything will be provided, without any input on your part.
switch off your mobile phone before
entering a laboratory. The main points to remember are:
●● Read any handouts in advance: make sure you understand the purpose
of the practical and the particular skills involved. Does the practical relate
to, or expand upon, a current topic in your lectures? Is there any additional
preparatory reading that will help?
Using textbooks in the lab – take this

●● Take along appropriate textbooks, to explain aspects in the practical.
book (or photocopies of relevant pages) ●● Consider what safety hazards might be involved, and any precautions
along to the relevant classes, so that you might need to take, before you begin (p. 6).
you can make full use of the informa-
tion during the practical sessions.
●● Listen carefully to any introductory guidance and note any important
points: adjust your schedule/handout, as necessary.
●● During the practical session, organise your bench space – make sure your
lab book is adjacent to, but not within, your working area. You will often find
it easiest to keep clean items of glassware, etc., on one side of your working
Safety Note If in doubt over any space, with used equipment on the other side.
part of the practical procedure – ASK!
There is no such thing as a silly ques-
●● All chemical waste (solid or liquid) should be disposed of in the
tion in the laboratory.
appropriate containers provided (consult the demonstrator or lecturer-
in-charge).
●● Write up your work as soon as possible and submit it on time or you may
lose marks.
●● Catch up on any work you have missed as soon as possible – preferably
before the next practical session.
Basic requirements
Presenting results – while you don’t Recording practical results
need to be a graphic designer to pro-
duce work of a satisfactory standard,
An A4 loose-leaf ring binder offers flexibility, since you can insert laboratory
presentation and layout are important
handouts, and lined and graph paper, at appropriate points. The danger of losing
and you will lose marks for poorly pre-
one or more pages from a loose-leaf system is the main drawback. Bound books
sented work.
avoid this problem, although those containing alternating lined/graph or lined/
blank pages tend to be wasteful – it is often better to paste sheets of graph paper
into a bound book, as required.

Essentials of practical work
All experimental observations and data should be recorded in a notebook

Presenting results – layout and pres-
in ink at the time they are made because it is easy to forget when you are busy.
entation of work are important. Ensure
A good-quality HB pencil or propelling pencil is recommended for making
that the information presented is legi-
diagrams, etc. as mistakes are easily corrected with a vinyl eraser. Buy a black,
ble. You will lose marks for poorly pre-
spirit-based (permanent) marker to label experimental glassware, sample tubes,
sented work. Chapter 6 gives further
etc. Fibre-tipped fine line drawing/lettering pens are useful for preparing final
practical advice.
versions of graphs and diagrams for assessment purposes. Use a clear ruler
(with an undamaged edge) for graph drawing, so that you can see data points/
information below the ruler as you draw.
Using calculators for numerical Calculators

problems – Chapter 6 gives further These range from basic machines with no pre-programmed functions and only
advice. one memory, to sophisticated programmable minicomputers with many mem-
ories. Note: Many university departments specify a particular make and model
of calculator for use in examinations. It is important that you purchase and
become familiar with the use of this calculator. The following may be helpful
when using a calculator:
●● Power sources. Choose a battery-powered machine, rather than a mains-
operated or solar-powered type. You will need one with basic mathematical/
scientific operations including powers, logarithms (p. 472), roots and paren-
Using calculators – take particular care theses (brackets), together with statistical functions such as sample means
when using the exponential key ‘EXP’ and standard deviations (Chapter 53).
or ‘EE’. Pressing this key produces
10 something. For example, if you want
●● Mode of operation. Calculators fall into two distinct groups. The older
to enter 2 * 10 -4 , the order entry is 2,
system used by, for example, Hewlett Packard calculators is known as the
EXP, - , 4 not 2, * , 10, EXP, - , 4.
reverse Polish notation: to calculate the sum of two numbers, the sequence
is 2 [enter] 4 + and the answer 6 is displayed. The more usual method of
calculating this equation is as 2 + 4 =, which is the system used by the
majority of modern calculators. Most newcomers find the latter approach
Using inexpensive calculators – many to be more straightforward. Spend some time finding out how a calculator
unsophisticated calculators have a operates, e.g. does it have true algebraic logic (U then number, rather than
restricted display for exponential num- number then U)? How does it deal with scientific notation (p. 471)?
bers and do not show the ‘power of 10’,
e.g. displaying 2.4 * 10-5 as 2.4-05, or
●● Display. Some calculators will display an entire mathematical operation
2.4E–05, or even 2.4–05.
(e.g. ‘2 + 4 = 6 ’), while others simply display the last number/operation.
The former type may offer advantages in tracing errors.
●● Complexity. In the early stages, it is usually better to avoid the more com-
plex machines, full of impressive-looking, but often unused preprogrammed
functions – go for more memory, parentheses or statistical functions rather
than engineering or mathematical constants. Programmable calculators may
be worth considering for more advanced studies. However, it is important to
note that such calculators are often unacceptable for exams.
Presenting more advanced practical work

In some practical reports and in project work, you may need to use more
sophisticated presentation equipment. Word processing may be essential and
Presenting graphs and diagrams –
computer-based graphics packages can be useful. Choose easily read fonts
ensure these are large enough to be
such as Arial or Times New Roman for project work and posters and consider
easily read: a common error is to pres-
the layout and content carefully (p. 601). Alternatively, you could use fine line
ent graphs or diagrams that are too
drawing pens plus dry-transfer lettering and symbols, such as those made by
small, with poorly chosen scales (see
Letraset®, although this approach is usually more time consuming and less
p. 453).
flexible than computer-based system, e.g. using Microsoft Excel.
4 The investigative approach

Essentials of practical work
The use of Microsoft PowerPoint® as a presentation package is common

Printing on acetates – standard over-
place. It is common to find a computer and presenter available for student use.
head transparencies are not suitable for
Advice on content and presentation is given in Chapter 68.
use in laser printers or photocopiers:
you need to make sure that you use the
correct type.
Source for further study

Bennett, S.W. and O’Neale, K. (1999) Progressive Devel- Overton, T., Johnson, S. and Scott, J. (2015) Study and
opment of Practical Skills in Chemistry. A guide to Communication Skills for the Chemical Sciences,
early-undergraduate experimental work. Royal Society 2nd edn., Oxford University Press, Oxford.
of Chemistry, Cambridge.
Study exercises
1.1 Consider the value of practical work. Spend a few 1.3 Check your calculator skills. Carry out the fol-
minutes thinking about the purpose of practical lowing mathematical operations, using either
work within a specific part of your course (e.g. a a hand-held calculator or a PC with appropriate
particular first year module) and then write a list of ‘calculator’ software.
the six most important points. Compare your list
(a) 5 * (2 + 6)
with the generic list we have provided on p. 602,
(b) [8.3 , (6.4 - 1.9)] * 24 (to 4 significant
which is based on our experience as lecturers –
figures)
does it differ much from your list, which is drawn
(c) (1 , 32) * (5 , 8) (to 3 significant figures)
up from a student perspective?
(d) 1.2 * 10 5 + 4.0 * 10 4 in scientific notation
1.2 Make a list of items required for a particular prac- (see p. 471)
tical experiment. This exercise is likely to be most (e) 3.4 * 10 -2 - 2.7 * 10 -3 in ‘normal’ notation
useful if you can relate it to an appropriate prac- (i.e. conventional notation, not scientific for-
tical session on your course. However, we have mat) and to 3 decimal places.
given a model list for a recrystallisation of an
impure compound from water as an example. (See also numerical exercises in Chapter 51)

2 Health and safety
Health and safety law requires institutions to provide a working environment

Health and Safety Legislation – in the
that is safe and without risk to health. Where appropriate, training and informa-
UK, the Health and Safety at Work Act
tion on safe working practices must be provided. Students and staff must take
1974 provides the main legal framework
reasonable care to ensure the health and safety of themselves and of others, and
for health and safety.The Control of Sub-
must not misuse any safety equipment.
stances Hazardous to Health (COSHH)
Regulations 2002 impose specific legal
requirements for risk assessment wher- Key Point All practical work must be carried out with safety
ever hazardous chemicals or biological in mind, to minimise the risk of harm to yourself and to others –
agents are used, with Approved Codes safety is everyone’s responsibility by law.
of Practice for the control of hazardous
substances, carcinogens and biological
agents, including pathogenic microbes. Risk assessment
A risk assessment is a systematic approach to hazard identification and control.
It is essential to consider what aspects of a laboratory activity can cause injury
to people and then to introduce control measures that will reduce the risk of
injury to an acceptable level. Important aspects to consider are:
●● Substance hazards
●● How the substance is to be used
Definitions
●● How it can be controlled
Hazard – the potential of a substance to
cause harm. ●● Who is exposed
Risk – the likelihood that a substance ●● How much exposure
will harm you and the severity of harm
in the actual circumstances of use. ●● The duration of exposure
Key Point It is important to distinguish between the HAZARD

of a substance and the RISK resulting from exposure.
The risk assessment process

The five step process requires you to:
1. Identify the hazards and risk: One way to do this is by using ‘PEME,’
i.e. People, Equipment, Materials and Environment.
a. ‘People’ hazards can cover a range of issues including the individual
themselves and the systems that people have to use. In this ‘people’
context consider the following terms: training, capabilities /restrictions,
supervision, communication, adequate numbers and human error.
b. ‘Equipment’ hazards relate to the equipment to be used; it will also
consider related aspects of the equipment including repair, mainte-
nance, handling, storage, cleaning and operation of the equipment.
c. ‘Materials’ hazards cover any liquid, solid or gas associated with
the task. This aspect also covers any by-products or waste generated
by the activity.

Health and safety
splash d. ‘Environment’ hazards relate to the surrounds you are working in.
inhalation
Examples include poor lighting, heating and ventilation, poor access
and egress, tripping/slipping hazards, restricted space/visibility and
ingestion
other activities taking place nearby.
2. Identify who can be harmed and how: Who – Athough a task may
intravenous
or seem to be well managed, if control measures fail then a whole range of
absorption people could be injured, e.g. co-lab workers in the area or people visiting
the area. Your risk assessment should consider all those people who could
potentially be harmed if the control measures fail. How – the five routes of
chemical exposure (Fig. 2.1) are: inhalation – breathing in small particles
or chemical vapours is the most common exposure pathway; dermal –
some chemicals can be absorbed into the body; ingestion – inadvertent
absorption
hand-to-mouth transmission; intravenously – improper use of needles/
(dermal) from glass pipettes and their disposal can lead to inadvertent exposure; eye
spillage contact – rubbing your eyes after chemical exposure with your hands (with
or without gloves).
Fig. 2.1 Major routes of entry of harmful 3. Identify the current controls and decide if more is required
substances into the body. a. Identify the control measures currently in place for each hazard
you have identified i.e. physical controls (i.e. local exhaust ventila-
tion); procedural controls (i.e. a safe working procedure for the task);
and behavioural controls (i.e. adequate supervision and monitoring of
Safety Note behaviour).
Protective clothing is worn as a first b. Identify the risks and decide on precautions – a risk matrix anal-
barrier to spillage of chemicals on to ysis. A risk analysis is a qualitative estimate of risk associated with
your body. each applicable risk; it assumes that the planned or existing controls
Lab coats are for protection of you and are in place. Box 2.1 shows you how to undertake a risk matrix anal-
your clothing. ysis. The risk matrix evaluates the risk by allocating a numeric risk
Eye protection special spectacles with level and the tolerability of the hazard.
side pieces to protect you from your
own mistakes and those of your col- 4. Record your findings – you will need to record your assessments. You
leagues. If you wear spectacles, eye will need to:
protection with prescription lenses a. state clearly what task /activity the risk assessment covers
and side pieces is available from your
optician, an expensive but worthwhile b. ensure that the hazards and controls are clearly listed
investment. Otherwise goggles can be
worn over spectacles. c. consider all those people who could potentially be harmed
Contact lenses should not be worn in d. ensure that the appropriate member of staff signs off the assessment
the laboratory. Chemicals can get under (e.g. technical demonstrator; lecturer-in-charge; project supervisor)
the lens and damage the eye before the
lens can be removed. It is often very dif- e. make sure the completed risk assessments are readily available to
ficult to remove the contact lens from those who might need them (e.g. module tutor).
the eye after a chemical splash.
5. Review as necessary. Risk assessments should be reviewed on a regular
Shoes should cover the feet: no open-
basis. The period of review should reflect the hazards: the greater the
toed sandals, for example.
hazards the more frequent the review. The risk assessments should also be
Long hair should be tied back and hats
reviewed, if for example, the experiment is modified in any way.
(e.g. baseball caps) should not be worn.

Health and safety
Box 2.1 How to perform a risk matrix analysis
A risk matrix analysis allows you to prioritise the likeli- environment / manner it is used in; in the absence of
hood and severity of risk to an individual from the hazard any specific control measures you should indicate the
identified. highest likelihood among the various risks (Table 2.3).
1. Using the form in Fig. 2.2 conduct a COSHH assess- 5. Assess the ‘severity’; this should be substance-spe-
ment of the chemical to be used in a practical labo- cific rather than activity-specific. This should relate
ratory class. If the Signal word is DANGER then the directly to the information provided on the MSDS
extended COSHH form should be used (Fig. 2.3). sheet (provided by the manufacturer); use the high-
est severity assessment among the various risks
2. First consult the Material Safety Data Sheet (MSDS) (Table 2.3).
supplied; all manufacturers of hazardous chemicals
are required to provide one of these sheets for all 6. Then, calculate the risk rating using the risk matrix
products which they sell. (Table 2.4). The risk is calculated by multiplying the
likelihood by the severity before any control meas-
3. Consult the Hazard pictograms (Fig. 2.4) for visible ures additional to Good Laboratory Practice (GLP)
relevant information. In addition, H (hazard) state- / Personal Protective Equipment (PPE) – laboratory
ments (Table 2.1) and P (precautionary) statements coat and safety glasses are factored in. This calcula-
(Table 2.2) are available on MSDS sheets and /or tion of risk should quote the highest risk associated
at: http://www.sigmaaldrich.com/help-welcome/ with the substance (i.e. what is the most dangerous
hazard-and-precautionary-statements.html. Enter feature of the substance).
the compound name in the search facility, then click
‘MSDS’ at the appropriate product line. 7. You are aiming to reduce the likelihood to as close to
1 as you can get (e.g. by performing the experiment
4. Assess the ‘likelihood’ of harm coming to pass in a fume cupboard).
given the amount/nature of substance used, the
An example MSDS sheet for phenol is shown in Fig. 2.5. In addition, an exam-
ple of a completed COSHH form for phenol is shown in Fig. 2.6. In addition,
as the Signal word is Danger an extended COSHH form (Fig. 2.3) would be
All manufacturers of hazardous chem-
required to be completed.
icals are required to provide a Material
Safety Data Sheet, or MSDS. The MSDS
Hazard statements – There are 72 individual and 17 combined Hazard state-
will contain the following information:
ments (Table 2.1). Each one of them is assigned a unique alphanumerical code
●● Manufacturer which consists of one letter and three numbers as follows:
●● Name of Chemical
●● Chemical Components ●● the letter ‘H’ (for ‘hazard statement’);
●● Hazards Associated with the Product ●● a number designating the type of hazard: ‘2’ for physical hazards; ‘3’ for
●● First Aid Measures
health hazards; and ‘4’ for environmental hazards; and finally,
●● Fire Fighting Measures
●● Handling and Storage ●● two numbers corresponding to the sequential numbering of hazards aris-
●● Accidental Release Procedures ing from the intrinsic properties of the substance or mixture, i.e. explosive
●● Exposure Control and Personal properties (codes from 200 to 210), flammability (codes from 220 to
Protection
230), etc.
●● Physical and Chemical Properties
●● Stability and Reactivity
●● Toxicological and Ecological Precautionary statements – There are 116 individual and 33 combined Pre-
Information cautionary statements (Table 2.2). These are assigned a unique alphanumerical
●● Disposal Practices code which consists of one letter and three numbers as follows:
●● Other miscellaneous information
●● the letter “P” (for ‘precautionary statement’);

Health and safety
●● one number designating the type of precautionary statement: ‘1’ for gen-
eral precautionary statements; ‘2’ for prevention precautionary statements;
‘3’ for response precautionary statements; ‘4’ for storage precautionary
statements; and, ‘5’ for disposal precautionary statements; and finally
●● two numbers (corresponding to the sequential numbering of precaution-
ary statements).
Fig. 2.2 Control of Substances Hazardous to Health (COSHH) form

Health and safety
Fig. 2.3 Extended Control of Substances Hazardous to Health (COSHH) form

Health and safety
Fig. 2.3 (continued)

Health and safety

Health and safety
Guidance notes for completing the COSHH form

1. H statements are available on MSDS sheets and/or at http://www.
sigmaaldrich.com/help-welcome/hazard-and-precautionary-statements.
html (Table 2.1) (enter compound name in search facility, click ‘MSDS’
at appropriate product line)
2. Each key/significant substance hazards identified on MSDS
3. If Signal word is DANGER then the Extended COSHH assessment
(Fig. 2.3) must be completed and the appropriate controls, storage, dis-
posal and emergency procedures transferred to this form. Provide as much
detail as is required to make this record suitable and sufficient
4. ‘Likelihood’ = Assessment of ‘likelihood’ of harm coming to pass given
the amount/nature of substance used, the environment/manner it’s used
in and in the absence of any specific Control Measures – indicate the
highest likelihood among the various risks
5. ‘Severity’: substance-specific rather than activity specific – indicated/
intimated on MSDS – use the highest severity assessment among the var-
ious risks.
6. Risk = Likelihood * Severity before control measures additional to
GLP (good lab practice)/PPE (Personal Protective Equipment = lab coat
and safety glasses are factored in). Note this column should quote the
highest risk associated with the substance (i.e. what is the most dangerous
feature of the substance).

Health and safety
Fig. 2.4 Hazard warning pictograms and their hazard codes

Table 2.1 Hazard statements
Signal
Code Hazard statements Hazard class Category word Pictogram P-Codes
H200 Unstable explosive Explosives Unstable Danger P201, P202, P281, P372, P373, P380, P401, P501
Explosive
H201 Explosive; mass explosion hazard Explosives Div 1.1 Danger P210, P230, P240, P250, P280, P370 +P380,
M02 Practical Skills in Chemistry 39920.indd 15

P372, P373, P401, P501
H202 Explosive; severe projection hazard Explosives Div 1.2 Danger P210, P230, P240, P250, P280, P370 +P380,
P372, P373, P401, P501
H203 Explosive; fire, blast or projection Explosives Div 1.3 Danger P210, P230, P240, P250, P280, P370 +P380,
hazard P372, P373, P401, P501
H204 Fire or projection hazard Explosives Div 1.4 Warning P210, P240, P250, P280, P370 +P380, P372,
P373, P374, P401, P501
H205 May mass explode in fire Explosives Div 1.5 Danger P210, P230, P240, P250, P280, P370 +P380,
P372, P373, P401, P501
Div 1.6
H220 Extremely flammable gas Flammable gases Category 1 Danger P210, P377, P381, P403
H221 Flammable gas Flammable gases Category 2 Warning P210, P377, P381, P403
H222 Extremely flammable aerosol Flammable aerosols Category 1 Danger P210, P211, P251, P410 +P412
H223 Flammable aerosol Flammable aerosols Category 2 Warning P210, P211, P251, P410 +P412
H224 Extremely flammable liquid and Flammable liquids Category 1 Danger
vapour
H225 Highly flammable liquid and Flammable liquids Category 2 Danger P210, P233, P240, P241, P242, P243, P280,
vapour P303 +P361 +P353, P370 +P378, P403 +P235,
P501
H226 Flammable liquid and vapour Flammable liquids Category 3 Warning
H227 Combustible liquid Flammable liquids Category 4 Warning P210, P280, P370 +P378, P403 +P235, P501
H228 Flammable solid Flammable solids Category 1 Danger P210, P240, P241, P280, P370 +P378
H228 Flammable solid Flammable solids Category 2 Warning
H240 Heating may cause an explosion Self-reactive sub- Type A Danger P210, P220, P234, P280,
stances and mix- P370 +P378, P370 +P380 +P375, P403 +P235,
tures; and Organic P411, P420, P501
peroxides
H241 Heating may cause a fire or Self-reactive sub- Type B Danger P210, P220, P234, P280,
explosion stances and mix- P370 +P378, P370 +P380 +P375, P403 +P235,
tures; and Organic P411, P420, P501
peroxides

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(continued)
10/05/2017 15:32
Table 2.1 (continued)
Signal
H242 Heating may cause a fire Self-reactive sub- Type C, D, Danger P210, P220, P234, P280,
stances and mix- P370 +P378, P403 +P235, P411, P420, P501

tures; and organic
peroxides
H242 Heating may cause a fire Self-reactive sub- Type E, F Warning P210, P220, P234, P280,
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stances and mix- P370 +P378, P403 +P235, P411, P420, P501
tures; and organic
peroxides

Type G
H250 Catches fire spontaneously if Pyrophoric liquids; Category 1 Danger P210, P222, P280, P302 +P334, P370 +P378,
exposed to air Pyrorophoric solids P422
H251 Self-heating; may catch fire Self-heating sub- Category 1 Danger

stances and mixtures
H252 Self-heating in large quantities; Self-heating sub- Category 2 Warning P235 +P410, P280, P407, P413, P420
may catch fire stances and mixtures
H260 In contact with water releases Substances and mix- Category 1 Danger
flammable gases which may ignite tures which, in con-
spontaneously tact with water, emit
flammable gases
H261 In contact with water releases flam- Substances and mix- Category 2 Danger P223, P231 +P232, P280,
mable gas tures which, in con- P335 +P334, P370 +P378, P402 +P404, P501
tact with water, emit
flammable gases
H261 In contact with water releases flam- Substances and mix- Category 3 Warning P231 +P232, P280, P370 +P378,
mable gas tures which, in con- P402 +P404, P501
tact with water, emit
flammable gases
H270 May cause or intensify fire; oxidiser Oxidising gases Category 1 Danger P220, P244, P370 +P376, P403
H271 May cause fire or explosion; strong Oxidising liquids; Category 1 Danger P210, P220, P221, P280, P283,
oxidiser Oxidising solids P306 +P360, P371 +P380 +P375, P370 +P378,
P501
H272 May intensify fire; oxidiser Oxidising liquids; Category 2 Danger
Oxidising solids
H272 May intensify fire; oxidiser Oxidising liquids; Category 3 Warning P210, P220, P221, P280, P370 +P378, P501
Oxidising solids
10/05/2017 15:32
H280 Contains gas under pressure; may Gases under Compressed Warning P410 +P403
explode if heated pressure gas
Liquefied gas
Dissolved
gas
H281 Contains refrigerated gas; may Gases under Refrigerated Warning P282, P336, P315, P403
cause cryogenic burns or injury pressure liquefied gas

H290 May be corrosive to metals Corrosive to metals Category 1 Warning P234, P390, P404
H300 Fatal if swallowed Acute toxicity, oral Category 1, 2 Danger P264, P270, P301 +P310, P321, P330, P405,
P501
H301 Toxic if swalloed Acute toxicity, oral Category 3 Danger P264, P270, P301 +P310, P321, P330, P405,
P501
H302 Harmful if swallowed Acute toxicity, oral Category 4 Warning P264, P270, P301 +P312, P330, P501
H303 May be harmful if swallowed Acute toxicity, oral Category 5 P312
H304 May be fatal if swallowed and Aspiration hazard Category 1 Danger

enters airways
H305 May be fatal if swallowed and Aspiration hazard Category 2 Warning P301 +P310, P331, P405, P501
enters airways
H310 Fatal in contact with skin Acute toxicity, dermal Category 1, 2 Danger P262, P264, P270, P280, P302 +P350, P310,
P322, P361, P363, P405, P501
H311 Toxic in contact with skin Acute toxicity, dermal Category 3 Danger P280, P302 +P352, P312, P322, P361, P363,
P405, P501
H312 Harmful in contact with skin Acute toxicity, dermal Category 4 Warning P280, P302 +P352, P312, P322, P363, P501
H313 May be harmful in contact with skin Acute toxicity, dermal Category 5 P312
H314 Causes severe skin burns and eye Skin corrosion/ Category 1A, Danger P260, P264, P280,
damage irritation B, C P301 +P330 +P331, P303 +P361 +P353, P363,
P304 +P340, P310, P321, P305 +P351 +P338,
P405, P501
H315 Causes skin irritation Skin corrosion/ Category 2 Warning P264 P280, P302 +P352, P321, P332 +P313,
irritation P362
H316 Causes mild skin irritation Skin c orrosion/ Category 3 Warning P332 +P313
irritation
H317 May cause an allergic skin reaction Sensitisation, skin Category 1 Warning P261, P272, P280, P302 +P352, P333 +P313,
P321, P363, P501

Health and safety
(continued)
10/05/2017 15:32
Table 2.1 (continued)
Signal
H318 Causes serious eye damage Serious eye damage/ Category 1 Danger P280, P305 +P351 +P338, P310
eye irritation

H319 Causes serious eye irritation Serious eye damage/ Category 2A Warning P264, P280, P305 +P351 +P338, P337 +P313P
eye irritation
Health and safety
H320 Causes eye irritation Serious eye damage/ Category 2B Warning P264, P305 +P351 +P338, P337 +P313
eye irritation
H330 Fatal if inhaled Acute toxicity, Category 1, 2 Danger P260, P271, P284, P304 +P340, P310, P320,

inhalation P403 +P233, P405, P501
H331 Toxic if inhaled Acute toxicity, Category 3 Danger P261, P271, P304 +P340 P311, P321,
inhalation P403 +P233, P405, P501
H332 Harmful if inhaled Acute toxicity, Category 4 Warning
inhalation
H333 May be harmful if inhaled Acute toxicity, Category 5 P261, P271, P304 +P340, P312 P304 +P312
inhalation
H334 May cause allergy or asthma symp- Sensitisation, Category 1 Danger P261, P285, P304 +P341, P342 +P311, P501
toms or breathing difficulties if respiratory
inhaled
H335 May cause respiratory irritation Specific target organ Category 3 Warning
toxicity, single expo-
sure; respiratory
tract irritation
H336 May cause drowsiness or dizziness Specific target organ Category 3 Warning P261, P271, P304 +P340, P312, P403 +P233,
toxicity, single expo- P405, P501
sure; Narcotic effcts
H340 May cause genetic defects Germ cell Category 1A, Danger
mutagenicity 1B
H341 Suspected of causing genetic Germ cat Category 2 Warning P201, P202, P281, P308 +P313, P405, P501
defects mutagenicity
H350 May cause cancer Carcinogenicity Category 1A, Danger

1B
H351 Suspected of causing cancer Carcinogenicity Category 2 Warning P201, P202, P281, P308 +P313, P405, P501
H360 May damage fertility or the unborn Reproductive toxicity Categoty 1A, Danger
child 1B
H361 Suspected of damaging fertility or Reproductive toxicity Category 2 Warning P201, P202, P281, P308 +P313, P405, P501
the unborn child
10/05/2017 15:32
H362 May cause harm to breast-fed Reproductive toxic- Additional P201, P260, P263, P264, P270, P308 +P313
children ity, effects on or via category
lactation
H370 Causes damage to organs Specific target Category 1 Danger P260, P264, P270, P307 +P311, P321, P405,
organ toxicity, single P501
exposure
H371 May cause damage to organs Specific target Category 2 Warning P260, P264, P270, P309 +P311, P405, P501

organ toxicity, single
exposure
H372 Causes damage to organs through Specific target organ Category 1 Danger P260, P264, P270, P314, P501
prolonged or repeated exposure toxicity, repeated
exposure
H373 Causes damage to organs through Specific target organ Category 2 Warning P260, P314, P501
prolonged or repeated exposure toxicity, repeated
exposure
H400 Very toxic to aquatic life Hazardous to the Category 1 Warning P273, P391, P501
aquatic environment,
acute hazard
H401 Toxic to aquatic life Hazardous to the Category 2 P273, P501

acute hazard
H402 Harmful to aquatic life Hazardous to the Category 3
acute hazard
H410 Very toxic to aquatic file with long Hazardous to the Category 1 Warning P273, P391, P501
lasting effects aquatic environment,
long-term hazard
H411 Toxic to aquatic life with long last- Hazardous to the Category 2
ing effects aquatic environment,
long-term hazard
H412 Harmful to aquatic life with long Hazardous to the Category 3
lasting effects aquatic environment,
long-term hazard
H413 May cause long lasting harmful Hazardous to the Category 4 P273, P501
effects to aquatic life aquatic environment,
long-term hazard
H420 Harms public health and the envi- Hazardous to the Category 1 Warning P502
ronment by destroying ozone in the ozone layer
upper atmosphere

Health and safety
10/05/2017 15:32
Health and safety
7. If the risk is low (1 to 10 inclusive) then only GLP and use of PPE is
required (this is standard for laboratory work). However, you may never-
theless deem additional control appropriate.
If risk is high (12 to 18 inclusive) then additional control measures
must be indicated.
If risk is very high (2 0 +) then this is probably not appropriate for
general undergraduate laboratory student activity, only trained per-
sonnel should be involved or demonstrations used.
Detail other control measures required to bring risk down to an
acceptable level, e.g. gloves, engineering (extraction), management
controls (SOPs, training).
Some specific hazard types require control even if risk is low, e.g.
highly toxic, carcinogen.
8. Controlled risk: additional control measures should reduce the ‘likelihood’
to 1 – the severity will be unchanged which means that this moderated risk
should now be the same number as the hazard severity.
9. P statements (Table 2.2) from the MSDS give advice on storage, precau-
tions during use, dealing with spillage, etc. for the concentrated materials.
10. Safe storage information relating to the form of the material actually in use
in the experiment.
11. List any key emergency procedures required in the event of spillage,
fire, etc.; this is for the material in the form used in the practical or
preparation.
12. Disposal must be through the correct waste route as specified by MSDS
or laboratory protocol. This information must be detailed enough that the
person actually disposing of the material or waste can do so both legally
and safely.
Table 2.2 Precautionary statements
Precautionary statements – General

P101 If medical advice is needed, have product container or label at hand.
P102 Keep out of reach of children.
P103 Read label before use.
Precautionary statements – Prevention

P201 Obtain special instructions before use.
P202 Do not handle until all safety precautions have been read and understood.
P210 Keep away from heat/sparks/open flames/hot surfaces – No smoking.
P211 Do not spray on an open flame or other ignition source.
P220 Keep/Store away from clothing/. . . /combustible materials.
P221 Take any precaution to avoid mixing with combustibles/. . .

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P222 Do not allow contact with air.

P223 Keep away from any possible contact with water, because of violent reaction and possible flash fire.
P230 Keep wetted with . . .
P231 Handle under inert gas.
P232 Protect from moisture.
P233 Keep container tightly closed.
P234 Keep only in original container.
P235 Keep cool.
P240 Ground/bond container and receiving equipment.
P241 Use explosion-proof electrical/ventilating/lighting/. . . /equipment.
P242 Use only non-sparking tools.
P243 Take precautionary measures against static discharge.
P244 Keep reduction valves free from grease and oil.
P250 Do not subject to grinding/shock/. . . /friction.
P251 Pressurised container: Do not pierce or burn, even after use.
P260 Do not breathe dust/fume/gas/mist/vapours/spray.
P261 Avoid breathing dust/fume/gas/mist/vapours/spray.
P262 Do not get in eyes, on skin, or on clothing.
P263 Avoid contact during pregnancy/while nursing.
P264 Wash hands thoroughly after handling.
P264 Wash skin thouroughly after handling.
P270 Do not eat, drink or smoke when using this product.
P271 Use only outdoors or in a well-ventilated area.
P272 Contaminated work clothing should not be allowed out of the workplace.
P273 Avoid release to the environment.
P280 Wear protective gloves/protective clothing/eye protection/face protection.
P281 Use personal protective equipment as required.
P282 Wear cold insulating gloves/face shield/eye protection.
P283 Wear fire/flame resistant/retardant clothing.
P284 Wear respiratory protection.
P285 In case of inadequate ventilation wear respiratory protection.
P231 + P232 Handle under inert gas. Protect from moisture.
P235 + P410 Keep cool. Protect from sunlight.
Precautionary statements – Response

P301 IF SWALLOWED:
P304 IF INHALED:
P305 IF IN EYES:
P306 IF ON CLOTHING:
P307 IF exposed:
P308 IF exposed or concerned:
(continued)

Health and safety
P309 IF exposed or if you feel unwell:

P310 Immediately call a POISON CENTER or doctor/physician.
P311 Call a POISON CENTER or doctor/physician.
P312 Call a POISON CENTER or doctor/physician if you feel unwell.
P313 Get medical advice/attention.
P314 Get medical advice/attention if you feel unwell.
P315 Get immediate medical advice/attention.
P320 Specific treatment is urgent (see . . . on this label).
P321 Specific treatment (see . . . on this label).
P322 Specific measures (see . . . on this label).
P330 Rinse mouth.
P331 Do NOT induce vomiting.
P332 IF SKIN irritation occurs:
P333 If skin irritation or rash occurs:
P334 Immerse in cool water/wrap in wet bandages.
P335 Brush off loose particles from skin.
P336 Thaw frosted parts with lukewarm water. Do not rub affected area.
P337 If eye irritation persists:
P338 Remove contact lenses, if present and easy to do. Continue rinsing.
P340 Remove victim to fresh air and keep at rest in a position comfortable for breathing.
P341 If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable for
breathing.
P342 If experiencing respiratory symptoms:
P350 Gently wash with plenty of soap and water.
P351 Rinse cautiously with water for several minutes.
P352 Wash with plenty of soap and water.
P353 Rinse skin with water/shower.
P360 Rinse immediately contaminated clothing and skin with plenty of water before removing clothes.
P361 Remove/Take off immediately all contaminated clothing.
P362 Take off contaminated clothing and wash before reuse.
P363 Wash contaminated clothing before reuse.
P370 In case of fire:
P371 In case of major fire and large quantities:
P372 Explosion risk in case of fire.
P373 DO NOT fight fire when fire reaches explosives.
P374 Fight fire with normal precautions from a reasonable distance.
P376 Stop leak if safe to do so. Oxidising gases (section 2.4) 1
P377 Leaking gas fire: Do not extinguish, unless leak can be stopped safely.
P378 Use . . . for extinction.
P380 Evacuate area.
P381 Eliminate all ignition sources if safe to do so.

Health and safety
P390 Absorb spillage to prevent material damage.

P391 Collect spillage. Hazardous to the aquatic environment
P301 + P310 IF SWALLOWED: Immediately call a POISON CENTER or doctor/physician.
P301 + P312 IF SWALLOWED: call a POISON CENTER or doctor/physician IF you feel unwell.
P301 + P330 + P331 IF SWALLOWED: Rinse mouth. Do NOT induce vomiting.
P302 + P334 IF ON SKIN: Immerse in cool water/wrap in wet bandages.
P302 + P350 IF ON SKIN: Gently wash with plenty of soap and water.
P302 + P352 IF ON SKIN: Wash with plenty of soap and water.
P303 + P361 + P353 IF ON SKIN (or hair): Remove/Take off immediately all contaminated clothing. Rinse SKIN with water/
shower.
P304 + P312 IF INHALED: Call a POISON CENTER or doctor/physician if you feel unwell.
P304 + P340 IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing.
P304 + P341 IF INHALED: If breathing is difficult, remove victim to fresh air and keep at rest in a position comfortable
for breathing.
P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy
to do. Continue rinsing.
P306 + P360 IF ON CLOTHING: Rinse immediately contaminated CLOTHING and SKIN with plenty of water before
removing clothes.
P307 + P311 IF exposed: Call a POISON CENTER or doctor/physician.
P308 + P313 IF exposed or concerned: Get medical advice/attention.
P309 + P311 IF exposed or if you feel unwell: Call a POISON CENTER or doctor/physician.
P332 + P313 IF SKIN irritation occurs: Get medical advice/attention.
P333 + P313 IF SKIN irritation or rash occurs: Get medical advice/attention.
P335 + P334 Brush off loose particles from skin. Immerse in cool water/wrap in wet bandages.
P337 + P313 IF eye irritation persists: Get medical advice/attention.
P342 + P311 IF experiencing respiratory symptoms: Call a POISON CENTER or doctor/physician.
P370 + P376 In case of fire: Stop leak if safe to Do so.
P370 + P378 In case of fire: Use . . . for extinction.
P370 + P380 In case of fire: Evacuate area.
P370 + P380 + P375 In case of fire: Evacuate area. Fight fire remotely due to the risk of explosion.
P371 + P380 + P375 In case of major fire and large quantities: Evacuate area. Fight fire remotely due to the risk of explosion.
Precautionary statements – Storage

P401 Store . . . .
P402 Store in a dry place.
P403 Store in a well-ventilated place.
P404 Store in a closed container.
P405 Store locked up.
P406 Store in corrosive resistant/. . . container with a resistant inner liner.
P407 Maintain air gap between stacks/pallets.
P410 Protect from sunlight.
P411 Store at temperatures not exceeding . . . °C/. . . °F.
(continued)

Health and safety
P412 Do not expose to temperatures exceeding 50 °C/ 122 °F.

P413 Store bulk masses greater than . . . kg/. . . lbs at temperatures not exceeding . . . °C/. . . °F.
P420 Store away from other materials.
P422 Store contents under . . .
P402 + P404 Store in a dry place. Store in a closed container.
P403 + P233 Store in a well-ventilated place. Keep container tightly closed.
P403 + P235 Store in a well-ventilated place. Keep cool.
P410 + P403 Protect from sunlight. Store in a well-ventilated place.
P410 + P412 Protect from sunlight. Do not expose to temperatures exceeding 50 °C/122 °F.
P411 + P235 Store at temperatures not exceeding . . . °C/. . . °F. Keep cool.
Precautionary statements – Disposal

P501 Dispose of contents/container to . . . .
P502 Refer to manufacturer/supplier for information on recovery/recycling.
Table 2.3 Assessing the likelihood and severity
Likelihood Severity
Certain / imminent 6 Multiple fatalities 6

Very likely 5 Single fatalities 5
Likely 4 Major injury 4
May occur 3 Lost time injury 3
Unlikely 2 Minor injury 2
Remote 1 Delay only 1
Table 2.4 Risk matrix
Risk: the risk in using the substance = Likelihood : Severity
LOW 1 to 10 Good laboratory practice required

HIGH 12 to 18 Specific identified control measures must be used
VERY HIGH 20 + Trained personnel only

Health and safety
Fig. 2.5 Example MSDS sheet for phenol

Health and safety

Health and safety

Health and safety

Health and safety

Health and safety

Health and safety

Health and safety

Health and safety
Fig. 2.6 Example Control of Substances Hazardous to Health (COSHH) form for phenol
Basic rules for laboratory work

●● Wear appropriate protective clothing at all times – a clean lab coat (buttoned
up), eye protection, appropriate footwear – and ensure your hair does not
constitute a hazard.
●● Never smoke, eat or drink in any laboratory, because of the risks of contam-
ination by inhalation or ingestion (Fig. 2.1).
●● Never work alone in a laboratory.
●● Make sure that you know what to do in case of fire, including exit routes,
how to raise the alarm, and where to gather on leaving the building. Remem-
ber that the most important consideration is human safety: do not attempt to
fight a fire unless it is safe to do so.
●● All laboratories display notices telling you where to find the first aid kit and
who to contact in case of accident/emergency. Report all accidents, even
those appearing insignificant – your department will have a reporting pro-
cedure to comply with safety legislation.
●● Know the hazard warning pictograms for specific chemical hazards (Figure 2.4).
●● Never touch chemicals unless they are known to have minimal hazard: use a
spatula to transfer and manipulate solids, and pipettes for liquids – see p. 63.

Health and safety
●● Never mouth pipette any liquid. Use a pipette filler (see p. 65).
●● Take care when handling glassware – see p. 68 for details.
●● Use a fume cupboard for hazardous chemicals. Make sure that it is working
and then open the front only as far as necessary: many fume cupboards are
marked with a maximum opening.
●● Always use the minimum quantity of any hazardous materials.
●● Work in a logical, tidy manner and minimise risks by thinking ahead.
●● Alway clear up spillages, especially around balances, infrared sample prepa-
ration areas, etc., for the next worker.
●● Always clear up at the end of each session. This is an important aspect of
safety, encouraging a responsible attitude towards laboratory work.
Sources for further study

Anon. (2003) Safety in Academic Chemistry Laboratories. Anon. (2016) Control of Substances Hazardous to Health
Accident prevention for college and university students, (COSHH). Health and Safety Executive. Available:
7th edn, Vol. 1 (Student copy). American Chemical www.hse.gov.uk/COSHH/inex.htm
Society. [Online access to COSHH including what the law
Anon. SIRI MSDS Index. Available: requires and advice on completing assessments.]
http://www.hazard.com/msds Last accessed 05/01/16.
Last accessed 05/03/16. Anon. (2016) Health and Safety Essentials. Learn Chemis-
[Online access to Materials Safety Data Sheets (MSDS) try, Royal Society of Chemistry. Available www.rsc.org/
for manufacturers’ chemicals]. learn-chemistry/collections health-and-safety.
Anon. (2010) Doing things safely is not just the right way Last accessed 05/01/16.
to work – it’s the only way! Safety for Introductory [Online access to information on: Accidents and Emer-
Chemistry Students. American Chemical Society. gencies; Chemical Safety; COSHH; Fire Safety in the
Laboratory; Laboratory Best Practices; Laboratory waste;
Anon. (2016) Chemical Health and Safety Resources.
Long Working; New and Expectant mothers; Risk Assess-
American Chemical Society. Available: www.acs.org/
ment; Safety Data Sheets; and, Workers with Disabilities.]
content/acs/en/education/policies/safety/chemical-
health-and-safety.html Day, R., Reader, J. A. and Rowland, E. (2003) Health,
[Online access to a range of resources.] Safety and Environmental Legislation. Royal Society of
Last accessed 05/01/16. Chemistry, Cambridge.
Study exercises
2.1 Test your knowledge of safe working proce- 2.2 Locate the relevant health and safety features in a
dures. After reading the appropriate sections of laboratory. Find each of the following in one of the
this book, can you remember the following: main laboratories used as part of your course (draw
a simple location map, if this seems appropriate):
(a) the four main steps involved in the process of
risk assessment; (a) fire exit(s);
(b) the major routes of entry of harmful sub- (b) fire-fighting equipment;
stances into the body; (c) first-aid kit;
(c) the warning labels for the major chemical haz- (d) eye-wash station;
ard symbols (either describe them or draw (e) waste flammable solvent container;
them from memory; (f) waste chlorinated solvent container;
(d) the international symbol for radioactivity? (g) broken glass container.

Health and safety
2.3 Investigate the health and safety procedures in 2.4 Carry out risk assessments for specific chemical
operation at your university. Can you find out the hazards. Look up the hazards associated with the
following: use of the following chemicals and list the appro-
priate protective measures required to minimise
(a) your university’s procedure in case of fire;
the risk during use in a lab class:
(b) the colour coding for fire extinguishers availa-
ble in your department and the recommenda- (a) ethanol to be used as a recrystallisation sol-
tions for use; vent for a solid (about 10 g);
(c) the accident reporting procedure used in your (b) sodium oxalate to be used to make a volumet-
department; ric standard solution (250.00 mL; 0.1 M);
(d) your department’s Code of Safe Practice relat- (c) sodium hydroxide, used in solid form to pre-
ing to project work? pare a dilute solution for neutralisation.

3 Making measurements
The term data (singular = datum, or data value) refers to items of information,
Definitions and you will use different types of data from a wide range of sources during
your practical work. Consequently, it is important to appreciate the underlying
Variable – any characteristic or property
features of data collection and measurement.
which can take one of a range of values
(contrast this definition with that for a
parameter, which is a numerical con- Variables
stant in any particular instance).
Variables (Fig. 3.1) can be classified as follows:
Statistic – an estimate of a parameter,
obtained from a sample (e.g. the height Quantitative variables
of 18-year-old females based on those
These are characteristics whose differing states can be described by means of
in your class).
a number. They are of two basic types:
●● Continuous variables, such as length; these are usually measured against a
numerical scale. Theoretically, they can take any value on the measurement
scale. In practice, the number of significant figures of a measurement is
directly related to the precision of your measuring system; for example,
Working with discontinuous variables – volumes of a pipette beaker measured with Vernier callipers will provide
note that while the original data values readings of greater precision than a millimetre ruler. Many of the variables
must be integers, derived data and sta- measured in sciences are continuous and quantitative, e.g. mass, temperature,
tistical values do not have to be whole time and amount of product formed by reaction.
numbers. Thus, it is perfectly acceptable
to express the mean number of children ●● Discontinuous (discrete) variables; these are always obtained by counting
per family as 2.4. and therefore the data values must be whole numbers (integers), with no
intermediate values.
Ranked variables
These provide data which can be listed in order of magnitude (i.e. ranked). A famil-
iar example is the abundance of an organism in a sample, which is often expressed
as a series of ranks, e.g. rare = 1, occasional = 2, frequent = 3, common = 4
and abundant = 5. When such data are given numerical ranks, rather than
descriptive terms, they are sometimes called ‘semi-quantitative data’. Note that
the difference in magnitude between ranks need not be consistent. For example,
VARIABLES
regardless of whether there was a one-year or a live-year gap between offspring
in a family, their ranks in order of birth would be the same.
quantitative Qualitative variables (attributes)
These are non-numerical and descriptive; they have no order of preference
continuous discontinuous ranked qualitative and, therefore, are not measured on a numerical scale nor ranked in order of
magnitude, but are described in terms of categories, e.g. the detection of the
presence or absence of a chemical by a colour test or precipitate.
height of number of order of sex of Variables may be independent or dependent. Usually, the variable under
children children birth of children
in a in a children
the control of the experimenter (e.g. time) is the independent variable, while
in a
family family in a family family the variable being measured is the dependent variable (p. 48). Sometimes it is
inappropriate to describe variables in this way and they are then referred to as
(1.3 m, (3, 0, 2) (1, 2, 3) (male,
0.653 m)
interdependent variables.
female)
The majority of data values are recorded as direct measurements, readings
Fig. 3.1 Examples of the different types of or counts, but there is an important group, called derived (or computed), which
variables as used to describe some charac- result from calculations based on two or more data values, e.g. ratios, percent-
teristics of families. ages, indices and rates.

Making measurements
Measurement scales
Variables may be measured on different types of scale:
●● Nominal scale: this classifies objects into categories based on a descriptive
characteristic. It is the only scale suitable for qualitative data.
●● Ordinal scale: this classifies by rank. There is a logical order in any number
scale used.
Example A nominal scale for temper- ●● Interval scale: this is used for quantitative variables. Numbers on an equal
ature is not feasible, since the relevant unit scale are related to an arbitrary zero point.
descriptive terms can be ranked in
order of magnitude. ●● Ratio scale: this is similar to the interval scale, except that the zero point now
represents an absence of that character (i.e. it is an absolute zero). In contrast
An ordinal scale for temperature
to the interval scale, the ratio of two values is meaningful (e.g. a temperature
measurement might use descriptive
terms, ranked in ascending order,
of 200 K is twice that of 100 K).
e.g. cold = 1, cool = 2, warm = 3, The measurement scale is important in determining the mathematical and
hot = 4. statistical methods used to analyse your data. Table 3.1 presents a summary of
The Celsius scale is an interval scale the important properties of these scales. Note that you may be able to meas-
for temperature measurement, since
ure a characteristic in more than one way, or you may be able to convert data
the arbitrary zero corresponds to the
freezing point of water (0°C).
collected in one form to a different form. For instance, you might measure
The Kelvin scale is a ratio scale for light in terms of the photon flux density between particular wavelengths of the
temperature measurement since 0 K EMR spectrum (ratio scale), or simply as ‘blue’ or ‘red’ (nominal scale); you
represents a temperature of absolute could find out the dates of birth of individuals (interval scale) but then use this
zero (for information, the freezing information to rank them in order of birth (ordinal scale). Where there are no
point of water is 273.15 K on this other constraints, you should use a ratio scale to measure a quantitative vari-
scale). able, since this will allow you to use the broadest range of mathematical and
statistical procedures (Table 3.1).
Table 3.1 Some important features of scales of measurement
Measurement scale
Nominal Ordinal Interval Ratio
Type of variable Qualitative (Ranked)* Ranked (Quantitative)* Quantitative Quantitative

(Quantitative)*
Examples Species Abundance scales Fahrenheit temperature Kelvin temperature
Sex Reproductive condition scale scale Weight Length
Colour Optical assessment of Date (BC/AD) Response time Most
colour development physical measurements
Mathematical Identity Identity Identity Magnitude Identity Magnitude
properties Magnitude Equal intervals Equal intervals True zero
point
Mathematical None Rank Rank Rank Addition Sub-
operations possible Addition Subtraction traction Multiplication
on data Division
Typical statistics used Only those based on Non-parametric Almost all types of Almost all types of
frequency of counts methods, sign tests. test, t-test, analysis test, t-test, ANOVA,
made: contingency Mann-Whitney U-test of variance (ANOVA), etc. (check distribution
tables, frequency etc. (check distribution before using, Chapter 53
distributions, etc. before using, Chapter 53 and 54)
Chi-square test and 54)
* In some instances (see text for examples).

Making measurements
Accuracy and precision

precise Accuracy is the closeness of a measured or derived data value to its true
but not value, while precision is the closeness of repeated measurements to each other
accurate
(Fig. 3.2), A balance with a fault in it (i.e. a bias, see below) could give precise
(i.e. very repeatable) but inaccurate (i.e. untrue) results. Unless there is bias
in a measuring system, precision will lead to accuracy and it is precision that
is generally the most important practical consideration, if there is no reason
accurate to suspect bias. You can investigate the precision of any measuring system by
but not repeated measurements of individual samples.
precise
Absolute accuracy and precision are impossible to achieve, due to both
the limitations of measuring systems for continuous quantitative data and the
fact that you are usually working with incomplete data sets (samples). It is
particularly important to avoid spurious accuracy in the presentation of results;
inaccurate
include only those digits that the accuracy of the measuring system implies.
and This type of error is common when changing units (e.g. inches to metres) and
imprecise
in derived data, especially when calculators give results to a large number of
decimal places. Further advice is given on p. 470.
Bias (systematic error) and consistency

precise
and
Bias is a systematic or non-random distortion and is one of the most trou-
accurate blesome difficulties in using numerical data. Biases may be associated with
incorrectly calibrated instruments, e.g. an electrode or syringe, or with exper-
imental manipulations, e.g. decomposition of a chemical during storage. Bias
Fig. 3.2 ‘Target’ diagrams illustrating preci-
in measurement can also be subjective, or personal, e.g. an experimenter’s
sion and accuracy. preconceived ideas about an ’expected’ result.
Bias can be minimised by using a carefully standardised procedure, with
fully calibrated instruments. You can investigate bias in ‘trial runs’ by measur-
ing a single variable in several different ways, to see whether the same result
is obtained.
To avoid personal bias, ‘blind’ measurements should be made where the
identity of individual samples is not known to the operator, e.g. using a coding
system.
Measurement error
All measurements are subject to error, but the dangers of misinterpretation are
reduced by recognising and understanding the likely sources of error and by
adopting appropriate protocols and calculation procedures.
A common source of measurement error is carelessness, e.g. reading a
scale in the wrong direction or parallax errors. This can be reduced greatly by
Minimising errors – determine early in careful recording and may be detected by repeating the measurement.
your study what the dominant errors Other errors arise from faulty or inaccurate equipment, but even a perfectly
are likely to be and concentrate your functioning machine has distinct limits to the accuracy and precision of its
time and effort on reducing these. measurements. These limits are often quoted in manufacturers’ specifications
and are applicable when an instrument is new; however, you should allow for
some deterioration with age. Further errors are introduced when the subject
being studied is open to influences outside your control. Resolving such prob-
Working with derived data – special
lems requires appropriate experimental design and representative sampling.
effort should be made to reduce meas-
One major influence virtually impossible to eliminate is the effect of the
urement errors because their effects
investigation itself: even putting a thermometer in a liquid may change the
can be magnified when differences,
temperature of the liquid. The very act of measurement may give rise to a
ratios, indices or rates are calculated.
confounding variable (p. 48), as discussed in Chapter 5. You should include

Making measurements
descriptions of possible sources of errors and estimates of their likely impor-

tance in any report and these should not be used as a catch-all excuse for poor
technique or inadequacies in your experimental design.

Anon. Suggestions for Keeping Laboratory Notebooks. Helmenstine, M.A. How to write a Lab Report. Available:
Available: http://otl.stanford.edu/inventors/resources/ http://www.chemistry.about.com
labnotebooks.html Zubrick, J.W. (2015) The Organic Chem Lab Survival
Last accessed 15/01/17. Manual. A student’s guide to techniques, 10th edn. John
[A Stanford University website which looks at the labo- Wiley & Sons Ltd, Chichester.
ratory notebook from a patent perspective]
Beavon, J.R.G. Writing the Laboratory Notebook. Avail-
able: http://www.home.dara/rod.beavon/lab_book.htm
Last accessed 15/01/17.
Study exercises
3.1 Classify variables. Decide on the type of varia- and obtained the readings shown in the table
bles used for the following measures, indicating alongside. Explain these results in terms of the
whether they are quantitative or qualitative, con- type of error involved in each case.
tinuous or discontinuous, and the type of scale
Companion of weights of masses on two balances
that would be used.
(a) time of a reaction Standard mass (g)

(b) titre of a titration 10 25 50 100 250
(c) percentage yield of a reaction Reading 10.050 25.049 50.051 100.048 250.052
(d) date of a sample (balance A)
3.2 Investigate types of error. A student weighed a Reading 10.004 25.011 50.021 100.039 250.102
set of standard masses on two electronic balances (balance B)
Answer to these study exercises are available at www.pearsoried.co.uk/practicalskills.

4 SI units and their use
When describing a measurement, you normally state both a number and a

Dimensionless measurements – some
unit (e.g. ‘the length is 1.85 metres’). The number expresses the ratio of the
quantities can be expressed as dimen-
measured quantity to a fixed standard, while the unit identifies that standard
sionless ratios or logarithms (e.g.
measure or dimension. Clearly, a single unified system of units is essential for
absorbance and pH), and in these cases
efficient communication of such data within the scientific community. The
you do not need to use a qualifying unit.
Système International d’Unités (SI) is the internationally ratified form of the
metre–kilogram–second system of measurement and represents the accepted
scientific convention for measurements of physical quantities.
Another important reason for adopting consistent units is to simplify com-
Table 4.1 The base and supplementary plex calculations where you may be dealing with several measured quantities
SI units
(see p. 468). Although the rules of the SI are complex and the scale of the base
Measured Name of
units is sometimes inconvenient, to gain the full benefits of the system you
quantity SI unit Symbol should observe its conventions strictly.
The description of measurements in SI involves:
Base units
●● seven base units and two supplementary units, each having a specified
Length metre m
abbreviation or symbol (Table 4.1);
Mass kilogram kg
Amount of substance mole mol
●● derived units, obtained from combinations of base and supplementary units,
which may also be given special symbols (Table 4.2);
Time second s
Electric current ampere A
●● a set of prefixes to denote multiplication factors of 103, used for convenience
to express multiples or fractions of units (Table 4.3).
Temperature kelvin K
Luminous intensity candela cd
Table 4.2 Some important derived SI units
Supplementary units
Plane angle radian rad Alternative
Solid angle steradian sr Definition in in derived
Measured quantity Name of unit Symbol base units units
Energy joule J m2 kg s -2 Nm
Force newton N m kg s -2 J m-1

Table 4.3 Prefixes used in the SI
Pressure pascal Pa kg m -1 s -2 N m-2
Multiple Prefix Symbol Multiple Prefix Symbol Power watt W m2 kg s -3 J s -1
10 -3 milli m 103 kilo k Electric charge coulomb C As J V-1
10 -6 micro m 106 mega M Electric potential
difference volt V m2 kg A-1s -3 J C-1
10 -9 nano n 109 giga G
Electric resistance ohm Ω m2 kg A-2 s -3 V A-1
10 -12 pico p 1012 tera T
Electric conductance siemens S s3 A2 kg-1m-2 A V-1 or Ω -1
10 -15 femto f 1015 peta P
Electric capacitance farad F s4A2 kg-1 m-2 C V-1
10 -18 atto a 1018 exa E
Luminous flux lumen lm cd sr
Illumination lux lx cd sr m-2 lm m-2
Frequency hertz Hz s -1
Example 10 mg is correct, while
Radioactivity becquerel Bq s -1
10mg, 10mg. and 10m g are incorrect;
2.6 mol is right, but 2.6 mols is wrong. Enzyme activity katal kat mol substrate s -1

SI units and their use
Recommendations for describing measurements in SI units

Basic format
●● Express each measurement as a number separated from its units by a
space. If a prefix is required, no space is left between the prefix and the unit
it refers to. Symbols for units are only written in their singular form and do
Example n stands for nano and
N for newtons.
not require full stops to show that they are abbreviated or that they are being
multiplied together.
●● Give symbols and prefixes appropriate upper or lower case ini-
tial letters as this may define their meaning. Upper case symbols are
Example 123456.789111 becomes
123 457.
named after persons but when written out in full they are not given initial
capital letters.
●● Show the decimal sign as a full point on the line. Some metric countries
continue to use the comma for this purpose and you may come across this
in the literature: commas should not therefore be used to separate groups
of thousands. In numbers that contain many significant figures, you should
separate multiples of 103 by spaces rather than commas.
Compound expressions for derived units

●● Take care to separate symbols in compound expressions by a space
to avoid the potential for confusion with prefixes. Note, for example, that
200 m s (metre seconds) is different from 200 ms (milliseconds).
●● Express compound units using negative powers rather than a solidus (/):
for example, write mol m -3 rather than mol/m3. The solidus is reserved for
separating a descriptive label from its units (see p. 451).
●● Use parentheses to enclose expressions being raised to a power if this
avoids confusion: for example J (k-1)mol-1.
●● Where there is a choice, select relevant (natural) combinations of derived
and base units, e.g. you might choose units of Pa m -1 to describe a hydro-
static pressure gradient rather than kg m -2s-1, even though these units are
equivalent and the measurements are numerically the same.
Use of prefixes
Examples 10 mm is preferred to 0.000 ●● Use prefixes to denote multiples of 103 (Table 4.3) so that numbers are
01 m or 0.010 mm.
kept between 0.1 and 1000.
1 mm2 = 1 * (10-3 m)2 = 1 * 10-6 m2
(not one-thousandth of a square metre). ●● Treat a combination of a prefix and a symbol as a single symbol. Thus,
3
1 dm (1 litre) is more properly when a modified unit is raised to a power, this refers to the whole unit includ-
expressed as 1 * (10-1m)3 = ing the prefix.
1 * 10-3 m3. ●● Avoid the prefixes deci (d) for 10−1, centi (c) for 10−2, deca (da) for 10
Avogadro’s constant is 6.022174 * and hecto (h) for 100 as they are not strictly SI.
1023 mol-1.
●● Express very large or small numbers as a number between 1 and 10
State as MW m -2 rather than W mm -2. multiplied by a power of 10 if they are outside the range of prefixes shown
in Table 4.3.
●● Do not use prefixes in the middle of derived units: they should be
attached only to a unit in the numerator (the exception is in the unit for
mass, kg).

Note – In this book, we use L and mL Key Point For the foreseeable future, you will need to make
where you would normally find equip- conversions from other units to SI units, as much of the litera-
ment calibrated in that way, but use SI ture quotes data using imperial, c.g.s. or other systems. You will
units where this simplifies calculations. need to recognise these units and find the conversion factors
In formal scientific writing, construc- required. Examples relevant to chemistry are given in Box 4.1.
tions such as 1 * 10-6m3 ( = 1 mL) and Table 4.4 provides values of some important physical constants
1 mm3 ( = 1 mL) may be used. in SI units.
Box 4.1 Conversion factors between some redundant units and the SI
Multiply number in old Multiply number in SI

unit by this factor for unit by this factor for
Quantity SI unit/symbol Old unit/symbol equivalent in SI unit* equivalent in old unit*
Area square metre/m2 acre 4.046 86 * 103 0.247 105 * 10 -3

hectare/ha 10 * 10 3
0.1 * 10 -3
square foot/ft 2 0.092 903 10.763 9
square inch/in2 645.16 * 10 -9 1.550 00 * 106
square yard/yd 2 0.836 127 1.195 99
Angle radian/rad degree/° 17.453 2 * 10 -3 57.295 8
Energy joule/J erg 0.1 * 10 -6 10 * 106

kilowatt hour/kWh 3.6 * 106 0.277 778 * 10 -6
Length metre/m Ångstrom/Å 0.1 * 10 -9 10 * 109
foot/ft 0.304 8 3.280 84
inch/in 25.4 * 10 -3 39.370 1
mile 1.609 34 * 103 0.621 373 * 10 -3
yard/yd 0.9144 1.093 61
Mass kilogram/kg ounce/oz 28.349 5 * 10 -3 35.274 0
pound/lb 0.453 592 2.204 62
stone 6.350 29 0.157 473
hundredweight/cwt 50.802 4 19.684 1 * 10 -3
ton (UK) 1.016 05 * 10 3
0.984 203 * 10 -3
Pressure pascal/Pa atmosphere/atm 101 325 9.869 23 * 10 -6
bar/b 100 000 10 * 10 -6
millimetre of mercury/mmHg 133.322 7.500 64 * 10 -3
torr/Torr 133.322 7.500 64 * 10 -3
Radioactivity becquerel/Bq curie/Ci 37 * 109 27.027 0 * 10 -12
Temperature kelvin/K centigrade (Celsius) degree/°C °C + 273.15 K - 273.15
Fahrenheit degree/°F (°F + 459.67) * 5/9 (K * 9/5) - 459.67
Volume cubic metre/m3 cubic foot/ft3 0.028 3168 35.314 7
3 -6
cubic inch/in 16.387 1 * 10 61.023 6 * 103
cubic yard/yd 3 0.764 555 1.307 95
UK pint/pt 0.568 261 * 10 -3 1 759.75
US pint/liq pt 0.473 176 * 10 -3 2 113.38
UK gallon/gal 4.546 09 * 10 -3 219.969
US gallon/gal 3.785 41 * 10 -3 264.172
*In the case of temperature measurements, use formulae shown.

Table 4.4 Some physical constants in SI terms

Converting between concentration
units – This is an important skill that Physical constant Symbol Value and units
should be practised.
Avogadro’s constant NA 6.022 174 * 1023 mol -1
Boltzmann’s constant k 1.380 626 JK-1
Charge of electron e 1.602 192 * 10 -19 C
Gas constant R 8.314 43 J K-1 mol -1
Faraday’s constant F 9.648 675 * 104 C mol -1
Molar volume of ideal gas at STP V0 0.022 414 * m3mol -1
Speed of light in vacuo c 2.997 924 * 108 ms -1
Planck constant h 6.626 205 * 10 -34 J s
Acceleration due to gravity g 9.807 m s -1
Atomic mass unit mu 1660 5402 * 10 -27 kg
Rydberg constant R∞ 1.097 * 107 m -1
Permitivity of vacuum e0 8.854 * 10 -12 F m-1
Some implications of SI in chemistry

Note – The other common non-SI
unit of volume is the cubic centimetre, Volume
cm3, (10 -2m)3. Even though they are The SI unit of volume is the cubic metre, m3, which is rather large for practical
not exactly the same, mL and cm3 are purposes. The litre (L) and the millilitre (mL) are technically obsolete, but are
used interchangeably, as are cubic dec- widely used and glassware is still calibrated using them. Note: The US spelling
imetre dm3 (10 -1m)3 = 10 -3 m3 and of litre is liter.
litre (L).
Mass
Conversion of non-SI units – the fol- The SI unit for mass is the kilogram (kg) rather than the gram (g): this is unu-
lowing conversions are useful when sual because the base unit has a prefix applied.
dealing with ‘traditional’ units:
1 L ≈ 1 dm3 = 1 * (10-1m)3 = Amount of substance

1 * 10-3 m3 You should use the mole (mol, i.e. Avogadro’s constant, see Table 4.4) to
1 mL = 1 * 10-3 * 10-3 m3 = express very large numbers. The mole gives the number of atoms in the atomic
1 * 1 * 10-6 m3 mass, a convenient constant.
1 mL = 1 * 10-6 * 10-3 m3 =
1 * 10-9 m3
Concentration
The SI unit of concentration, mol m -3, is not convenient for general laboratory
work. It is equivalent to the non-SI term ‘millimolar’ (mM K mmol L-1) while
A Unit oddity – An example of nomen- ‘molar’ (M K mol L-1) becomes kmol m -3. If the solvent is not specified, then
clature that does not fit easily into the it is assumed to be water (see Chapter 11).
SI system.
The UK convention is 10/03/17 for the Time
10th March 2017, but in the USA this
would mean the 3rd October 2017. Be
In general, use the second (s) when reporting physical quantities having a time
clear which convention you are adopt-
element. Hours (h), days (d) and years should be used if seconds are clearly
ing for the date.
absurd (e.g. samples were taken over a 5-year period). Note, however, that you
may have to convert these units to seconds when doing calculations.

Box 4.2 How to interconvert SI units
Example: You are required to calculate the molecular 2. Look at the units and decide which are common:
weight of a polymer by measurements of its osmotic since the gas constant is expressed in joules, you
pressure in solution. At infinite dilution, measured should convert the osmotic pressure term into joules.
graphically from your experiments, the equation below The derived unit for pressure is N m -2 and, since the
applies: derived units for N are J m -1, the full derived unit of
pressure is (J m -1) * m -2 = J m -3.
Π RT
=
c Mr 3. Substitute the units into the equation for M:
where Π = osmotic pressure at infinite dilution (Pa), RTc J K -1 mol-1 * K * kg m -3
Mr = = = kg mol-1
R = gas constant (J K -1mol-1), T = temperature (K), Π J m -3
c = concentration of solution (kg m -3) and Mr = molecular
weight. 4. Substitute the appropriate numerical values into the
equation for Mr: you know that the units of the cal-
1. Re-arrange the equation for Mr: culation will be correct since the molecular weight is
the weight of 1 mole of polymer, expressed in kg.
RTc
Mr =
Π
Temperature
Definitions
The SI unit is the kelvin, K. The degree Celsius scale has units of the same
STP - Standard Temperature and magnitude, °C, but starts at 273.15 K, the melting point of ice at STP. Temper-
Pressure = 293.15 K and 101325Pa (or ature is similar to time in that the Celsius scale is in widespread use, but note
101.325 kPa or 0.101 325 M Pa). that conversions to K may be required for calculations. Note also that you must
not use the degree sign (°) with K and that this symbol must be in upper case
to avoid confusion with k for kilo; however, you should retain the degree sign
with °C to avoid confusion with the coulomb, C.
Interconversion of SI units
You will find that the use of SI units simplifies mathematical manipulations
and ensures that you obtain the correct units for the parameter being calculated.
Remember that you must convert all units into the appropriate SI units, e.g.
masses must be expressed as kg, volumes as m3 and concentrations as kg m -3
or mol m -3, etc., and that you may need to use alternatives in derived units
(Table 4.2). The application of these principles is shown in Box 4.2.

Anon. (2000) The NIST Reference on Constants, Units and Anon. (2014). Measurement units: the SI. Available:
Uncertainty. Available: http://physics.nist.gov/cuu/units/ www.bipm.org/en/measurement-units/
index.html Last accessed 05/01/16.
Last accessed 05/01/16. [Online information includes the SI brochure: The
Anon. Measurement units. Available: www.npl.co.uk/ International System of Units (SI), 9th edn (draft).]
reference/measurement-units/ Blackman, A. and Gahan, L. (2014) Aylward and Findlay’s
Last accessed 05/01/16. SI Chemical Data, 7th edn. John Wiley & Sons Ltd,
Chichester.

Study exercises
4.1 Practise manipulation of equations and deduction A = absorbance (dimensionless);

of SI base units in the following examples using c = concentration (mol L-1);
the procedures outlined in Box 4.2. l = length (cm); e = molar absorptivity (?)
(a) PV = nRT 1 k
P = pressure (kg m -1s-2): V = volume (m3); (c) v =
2p A m
n = number of moles (mol); v = frequency (s-1); m = reduced mass (kg);
T = temperature (K); R = gas constant (?). p = 3.142
(b) A = ecl k = force constant (?)

5 Scientific method and design of
experiments
Science is a body of knowledge based on observation and experiment. Scien-

Definitions tists attempt to explain their observations in terms of theories and hypotheses.
They make predictions from these hypotheses and test them by experiment or
Paradigm. Theoretical framework so
further observations. The philosophy and sociology that underlie this process
successful and well-confirmed that
are complex topics. Any brief description must involve simplifications.
most research is carried out within its
context and doesn’t challenge it – even Figure 5.1 models the scientific process you are most likely to be involved
significant difficulties can be ‘shelved’ in in testing ‘small-scale’ hypotheses. These represent the sorts of explanation
favour of its retention. that can give rise to predictions which can be tested by an experiment or a
Theory. A collection of hypotheses that series of observations. For example, you might put forward the hypothesis that
covers a range of natural phenomena – the rate of loss of a substance from a sample is dependent on temperature. An
a ‘larger-scale’ idea than a hypothesis. experiment could be set up to test this hypothesis, and the results would either
Note that a theory may be ‘hypotheti- confirm or refute the hypothesis.
cal’, in the sense that it is a tentative If confirmed, a hypothesis is retained with greater confidence. If refuted,
explanation. it is either rejected outright as false, or modified and retested. Alternatively, it
Hypothesis. An explanation tested in a might be decided that the experiment was not a valid test of the hypothesis –
specific experiment or by a set of obser- perhaps because it was later found that the substance was sample dependent.
vations. Tends to involve a ‘small scale’ Nearly all scientific research deals with the testing of small-scale hypoth-
idea. eses. These hypotheses operate within a theoretical framework that has proven
(Scientific) Law. This concept can be to be successful i.e. is confirmed by many experiments and is consistently
summarised as an equation (law) that predictive. This operating model or ‘paradigm’ is not changed readily, and, even
provides a succinct encapsulation of a
if a result appears that seems to challenge the conventional view, would not
system, often in the form of a mathe-
be overturned immediately. The conflicting result would be ‘shelved’ until an
matical relationship. The term is often
used in the physical sciences (e.g. explanation was found after further investigation. In the example used above, a
‘Beer’s Law’, p. 217). relevant paradigm could be the notion that life processes are ultimately chem-
ical in nature.
Although changes in paradigms are rare, they are important, and the sci-
entists who recognise them become famous. Generally, however, results from
hypothesis-testing tend to support and develop (‘articulate’) this paradigm,
Paradigm enhancing its relevance and strengthening its status.
Where do ideas for small-scale hypotheses come from? They arise from
one or more thought processes on the part of a scientist:
Articulation
Observations, ●● analogy with other systems;
techniques,
Hypothesis
concepts and ●● recognition of a pattern;
ideas
Prediction
and ●● recognition of departure from a pattern;
testing
●● invention of new analytical methods;
Observations and
experiments
●● development of a mathematical model;
Fig. 5.1 A model of scientific method as ●● intuition;
used when testing hypotheses on a small
scale. Hypotheses can arise as a result of ●● imagination.
various thought-processes on the part of Recently, it has been recognised that the process of science is not an entirely
the scientist, and are consistent with the
objective one. For instance, the choice of analogy which led to a new hypoth-
overlying paradigm. Each hypothesis is
testable by experiment or observation,
esis might well be subjective, depending on past knowledge or understanding.
leading to its confirmation or rejection. Also, science is a social activity, where researchers put forward and defend
Confirmed hypotheses act to strengthen viewpoints against those who hold an opposing view; where groups may work
the status of the paradigm, but rejected together towards a common goal; and where effort may depend on externally
ones do not immediately result in the par- dictated financial opportunities and constraints. As with any other human activ-
adigm’s replacement. ity, science is bound to involve an element of subjectivity.

Scientific method and design of experiments
How are decisions made about whether to accept or reject a hypothesis?

This is sometimes clear-cut, where experiments can be set up to result in a
Deciding whether to accept or reject a
binary outcome. In many other cases, the existence of ‘natural variation’ means
hypothesis – this is sometimes clear-
that statistical techniques need to be employed (Chapters 53 and 54).
cut, as in some areas of genetics where
No hypothesis can ever be rejected with certainty. Statistics might allow us
experiments can be set up to result in a
to quantify as vanishingly small the probability of an erroneous conclusion, but
binary outcome. In many other cases,
we are nevertheless left in the position of never being 100% certain that we have
the existence of ‘chemical variation’
rejected all relevant alternative hypotheses, nor 100% certain that our decision to
means that statistical techniques need
reject some alternative hypotheses was correct! However, despite these problems,
to be employed (Chapters 53 and 54).
experimental science has yielded and continues to yield many important findings.
Key Point The fallibility of scientific ‘facts’ is essential to

grasp. No explanation can ever be 100% certain as it is always
possible for a new alternative hypothesis to be generated. Our
understanding of science changes all the time as new observa-
tions and methods force old hypotheses to be retested.
Quantitative hypotheses, those involving a mathematical description of the

system, have become very important, because they can be formulated concisely
by mathematical models. Formulating models is often useful because it forces
deeper thought about mechanisms and encourages simplification of the system.
A mathematical model:
●● is inherently testable through experiment;
Definition ●● identifies areas where information is lacking or uncertain;

●● encapsulates many observations;
Mathematical model. An algebraic
summary of the relationship between ●● allows you to predict the behaviour of a system.
the variables in a system.
Remember, however, that assumptions and simplifications required to create a
model may result in it being unrealistic. Further, the results obtained from any
model are only as good as the information put into it.
The terminology of experimentation

field (whole area blocks (subdivisions
available)
In many experiments, the aim is to provide evidence for causality. If x causes y,
of field)
we expect, repeatably, to find that a change in x results in a change in y. Hence,
the ideal experiment of this kind involves measurement of y, the dependent
(measured) variable, at one or more values of x, the independent variable, and
subsequent demonstration of some relationship between them. Experiments
therefore involve comparisons of the results of treatments – changes in the
independent variable as applied to an experimental subject. The change is engi-
neered by the experimenter under controlled conditions.
Subjects given the same treatment are known as replicates. A ‘block’ is a
grouping of replicates. The blocks are contained in a ‘field’, i.e. the whole area
(or time) available for the experiment (Fig. 5.2). These terms originated from
the statistical analysis of agricultural experiments, but they are now used for
replicates within block = plots
many other areas of science.
Fig. 5.2 Terminology and physical arrange-
ment of elements in an experiment. Each Why you need to control variables in experiments
block should contain the complete range of
treatments (treatments may be replicated Interpretation of experiments is seldom clear-cut because uncontrolled varia-
more than once in each block). bles always change when treatments are given.

Confounding variables
These increase or decrease systematically as the independent variable increases
or decreases. Their effects are known as systematic variation. This form of
variation can be disentangled from that caused directly by treatments by
incorporating appropriate controls in the experiment. A control is really just
another treatment where a potentially confounding variable is adjusted so that
its effects, if any, can be taken into account. The results from a control may
therefore allow an alternative hypothesis to be rejected. There are often many
potential controls for any experiment.
The consequence of systematic variation is that you can never be certain
that the treatment, and the treatment alone, has caused an observed result, espe-
cially when using biological material. By careful design you can, however, ‘min-
imise the uncertainty’ involved in your conclusion. Methods available include:
●● Ensuring, through experimental design, that the independent variable is the
only major factor that changes in any treatment.
●● Incorporating appropriate controls to show that potential confounding vari-
ables have little or no effect.
●● Selecting experimental subjects randomly to cancel out systematic variation
arising from biased selection.
●● Matching or pairing individuals among treatments so that differences in
response due to their initial status are eliminated.
●● Arranging subjects and treatments randomly so that responses to systematic
differences in conditions do not influence the results.
Reducing edge effects – One way to do ●● Ensuring that experimental conditions are uniform so that responses to
this is to incorporate a ‘buffer zone’ of systematic differences in conditions are minimised. When attempting this,
untreated subjects around the experi- beware of ‘edge effects’ where subjects on the periphery of the layout expe-
ment proper. rience substantially different conditions from those in the centre.
Nuisance variables
These are uncontrolled variables which cause differences in the value of y
independently of the value of x, resulting in random variation. Experimental
science is characterised by the high number of nuisance variables that are found
and their relatively great influence on results – some chemical data tend to
have large errors! To reduce and assess the consequences of nuisance variables:
●● incorporate replicates to allow random variation to be quantified;
●● choose subjects that are as similar as possible;
●● control random fluctuations in environmental conditions.
Constraints on experimental design

Box 5.1 outlines the important stages in designing an experiment. At an early
Evaluating design constraints – a good stage, you should find out how resources may constrain the design. For exam-
way to do this is by processing an indi- ple, limits may be set by availability of subject materials, cost of treatment,
vidual subject through the experimental availability of a chemical or bench space. Logistics may be a factor (e.g. time
procedures – this type of ‘preliminary taken to record or analyse data).
run’ can help to identify potential Your equipment or facilities may affect design because you cannot regulate
difficulties. conditions as well as you might desire. For example, you may be unable to ensure
that the temperature is constant over an experiment laid out in an open laboratory.

Box 5.1 Checklist for designing and performing an experiment
1. Preliminaries 3. Planning
(a) Read background material and decide on a sub- (a) List all the materials you will need – order any
ject area to investigate. chemicals and make up solutions; obtain the
(b) Formulate a simple hypothesis to test – it experimental material you require; check equip-
is preferable to have a clear answer to one ment is available.
question than to be uncertain about several (b) Organise space and/or time in which to do the
questions. experiment.
(c) Decide which dependent variable you are going (c) Account for the time taken to apply treatments
to measure and how – is it relevant to the prob- and record results – make out a timesheet if
lem? Can you measure it accurately, precisely things will be hectic.
and without bias?
4. Carrying out the experiment
(d) Think about and plan the statistical analysis of
your results – will this affect your design? (a) Record the results and make careful notes of
everything you do (see p. 601) – make additional
observations to those planned if interesting things
2. Designing happen (see Resources section; Bird et al., 2013).
(b) Repeat the experiment if time and resources
(a) Find out the limitations on your resources.
allow.
(b) Choose treatments and conditions that alter the
minimum of confounding variables. 5. Analysing
(c) Incorporate as many effective controls as (a) Graph data as soon as possible (during the
possible. experiment if you can) – this will allow you to
(d) Keep the number of replicates as high as is visualise what has happened and make adjust-
feasible. ments to the design, if this seems appropriate
(e) Ensure that the same number of replicates is (e.g. timing of measurements).
present in each treatment. (b) Carry out the planned statistical analysis.
(f) Use effective randomisation and blocking (c) Jot down conclusions and new hypotheses
arrangements. arising from the experiment.
Using replicates
Replicate results show how variable the response is within treatments. They
Deciding the number of replicates in allow you to compare the differences among treatments in the context of the
each treatment – try to maximise the variability within treatments – you can do this via statistical tests such as anal-
number of replicates in each treat- ysis of variance (Chapter 53). Larger sample sizes tend to increase the precision
ment within the constraints of time and of estimates of parameters and increase the chances of showing a significant
resources available, using the same difference between treatments, if one exists. For statistical reasons (weighting,
number of replicates for each treatment. ease of calculation, fitting data to certain tests), it is often best to keep the num-
ber of replicates even. Remember that the degree of independence of replicates
is important – sub-samples cannot act as replicate samples, they tell you about
variability in the measurement method but not in the quantity being measured.
If the total number of replicates available for an experiment is limited by
resources, you may need to compromise between the number of treatments and
Definition the number of replicates per treatment. Statistics can help here, for it is pos-
sible to work out the minimum number of replicates you would need to show
Interaction. Where the effect of treat-
a certain difference between pairs of means (say 10%) at a specified level of
ments given together is greater or less
significance (say P = 0.05). For this, you need to obtain a prior estimate of
than the sum of their individual effects.
variability within treatments.

2
Factor B
1
Multi-factorial experiments
The simplest experiments are those in which one treatment (factor) is applied
2 at a time to the subjects. This approach is likely to give clear-cut answers, but
0 b
it could be criticised for lacking realism. In particular, it cannot take account of
interactions among two or more conditions that are likely to occur in real life.
Factor
A A multi-factorial experiment (Fig. 5.3) is an attempt to do this the interactions
among treatments can be analysed by specialised statistics.
1 a a1b1c Multi-factorial experiments are economical on resources because of
‘hidden replication’. This arises when two or more treatments are given to a
subject because the result acts statistically as a replicate for each treatment.
Choice of relevant treatments to combine is important in multi-factorial exper-
Fig. 5.3 Design of a simple multi-factorial
experiment. Factors A and B have effects a
iments for instance, an interaction may be present at certain concentrations
and b when applied alone. When both are of a chemical but not at others (perhaps because the response is saturated).
applied together, the effect is denoted by It is also important that the measurement scale for the response is consistent,
a + b + c. otherwise spurious interactions may occur. Beware when planning a multi-
factorial experiment that the number of replicates do not get out of hand – you
●● If c = 0, there is no interaction (e.g.
may have to restrict the treatments to ‘plus’ or ‘minus’ the factor of interest
2 + 2 + c = 4).
●● If c is positive, there is a positive interac-
(as in Fig. 5.3).
tion (synergism) between A and B (e.g.
2 + 2 + c = 5). Repetition of experiments
●● If c is negative, there is a negative inter-
action (antagonism) between A and B Even if your experiment is well designed and analysed, you should understand
(e.g. 2 + 2 + c = 3). that only limited conclusions can be made. Firstly, what you can say is valid for
a particular place and time, with a particular investigator, experimental subject
Reporting correctly – it is good prac- and method of applying treatments. Secondly, if your results were significant
tice to report how many times your at the 5% level of probability (p. 492), there is still an approximately one-
experiments were repeated (in Mate- in-twenty chance that the results did arise by chance. To guard against these
rials and Methods). In the Results possibilities, it is important that experiments are repeated. Ideally, this would
section, you should add a statement be done by an independent scientist with independent materials. However, it
saying that the illustrated experiment makes sense to repeat work yourself so that you can have full confidence in
is representative. your conclusions. Many scientists recommend that experiments be done three
times in total, but this may not be possible in undergraduate work.
Text reference
Bird, C. L., Willough by, C. and Frey, J. G. (2013) Labo- record keeping for chemistry and other sciences. Chem.
ratory notebooks in the digital era: the role of ELNs in Soc. Rev. 42, 8157–8175.
For further study

Zubrick, J.W. (2015) The Organic Chem Lab Survival
Manual. A student’s guide to techniques, 10th edn. John
Wiley & Sons Ltd, Chichester.

Study exercises
5.1 Generate random numbers. Produce a list of 20 test formula(e) to several cells to test empirically
random whole numbers between 1 and 5 using whether it works.
a spreadsheet. If using Excel, investigate the 5.2 Design and plan an experiment. Using one of
RAND( ) and INT functions. The RAND( ) function your practicals, determine the purpose of the
produces a random number between 0 and 1, so experiment and what it aims to achieve. List any-
you will need to multiply by a constant factor to thing else you could do to make the results more
scale your final output appropriately. Copy your robust.
Resources
Hardbounds books: examples Mitchells Laboratory Note- labfolder.com. Available: www.labfolder.com
books. Avaliable: www.laboratorynotebooks.co.uk Last accessed 05/01/16.
Last accessed 05/01/16. labarchives.com. Available: www.labarchives.com
Eureka Lab Book. Available: www.eurekalabbook.com Last accessed 05/01/16.
Last accessed 05/01/16. Marsfield, N. (2007). Key Skills for scientists. Getting the
Electronic Lab Notebooks (ELNs): examples cambridge- message across. Section 2 Keeping a laboratory note-
soft.com. Available: www.cambridgesoft.com book, Royal Society of Chemistry, Cambridge.

6 Making notes of practical work
When carrying out lab work or research projects, you will need to master
the important skills of recording and managing data. Individual observations
(e.g. laboratory temperature) can be recorded in the text of your notes, but
tables are the most convenient way to collect large amounts of information.
●● An accurate and neat record helps when using information later, perhaps for
exam purposes or when writing a report.
Understanding what’s expected – espe-
●● It allows you to practise important skills such as scientific writing, drawing
cially when taking notes for a lab-based
diagrams, preparing graphs and tables and interpreting results.
practical, pay special attention to the
aims and learning objectives (p. 551) ●● Analysing and writing up your data as you go along prevents a backlog at
of the session, as these will indicate the end of your study time.
the sorts of notes you should be taking,
●● You can show your work to a future employer to prove you have developed
including content and diagrams, and the
the skills necessary for writing up properly; in industry, this is vital so that
ways in which you should present these
others in your team can interpret and develop your work.
for assessment.
Key Point A good set of lab notes should:

Recording primary data – never be
●● outline the purpose of your experiment or observation;
tempted to jot down data on scraps of
●● set down all the information required to describe your exper-
paper: you are likely to lose them, or to
imental section;
forget what individual values mean.
●● record all relevant information about your results or observa-
tions and provide a visual representation of the data;
Means of recording notes – the tra-
●● note your immediate conclusions and suggestions for further
ditional paper-based bound notebook
experiments.
is still favoured by many, and has the
advantage of reliability and portability. Collecting and recording primary data
Increasingly, laptop and notebook com-
When preparing a table for data collection, you should:
puters are used; and an advantage here
is that recorded data can be transferred 1. Use a concise title or a numbered code for cross-referencing.
easily between applications and asso-
2. Decide on the number of variables to be measured and their relationship
ciated files for analysis. In both cases,
with each other and lay out the table appropriately:
backup of data is essential.
(a) The first column of your table should show values of the independ-
ent (controlled) variable, with subsequent columns for the individual
(measured) values for each replicate or sample.
Labelling your notes – always add a (b) If several variables are measured for the same sample, each should be
date (and time) to each of your primary given a row.
record files.
(c) In time-course studies, put the replicates as columns grouped accord-
ing to treatment, with the rows relating to different times.
3. Make sure the arrangement reflects the order in which the values will
be collected. Your table should be designed to make the recording process
as straightforward as possible, to minimise the possibility of mistakes. For
final presentation, a different arrangement may be best (Chapter 50).
Designing a table for data collection –
make sure there is sufficient space in
4. Consider whether additional columns are required for subsequent
each column for the values; if in doubt,
calculations. Create a separate column for each mathematical manipula-
err on the generous side.
tion, so the step-by-step calculations are clearly visible. Use a computer
spreadsheet (p. 425) if you are manipulating lots of data.

Making notes of practical work
5. Take sufficient time to record quantitative data unambiguously –

Recording numerical data – write
use large, clear numbers, making sure that individual numerals cannot be
down only those numbers that can be
confused.
justified by your measurement tech-
nique (significant figures). 6. Record numerical data to an appropriate number of significant figures,
reflecting the accuracy and precision of your measurement (p. 38). Do not
round off data values, as this might affect the subsequent analysis.
7. Record the actual observations, not your interpretation, e.g. the colour
of a particular chemical test, rather than whether the test was positive or
negative. Take care not to lose any of the information content of the data:
for instance, if you only write down means and not individual values, this
will affect your ability to carry out subsequent statistical analyses.
8. Prepare duplicated recording tables/checklists for repeated
experiments.
9. Explain any unusual results in a footnote. Don’t rely on memory.
Recording details of project work

The recommended system is one where you make a dual record.
Primary record
The primary record is made at the bench and you must concentrate on the detail
Tips for recording notes on pages:
of materials, methods and results. Include information that would not be used
●● use a refill pad or spiral-band note-
book, so you can easily deal with
elsewhere, but which might prove useful in error tracing: for example, if you
mistakes. note how a solution was made up (exact volumes and weights used rather than
●● If you need to use loose paper, make concentration alone), this could reveal whether a miscalculation had been the
sure each sheet is dated and num- cause of a rogue result. Note the origin, type and state of the chemicals used.
bered (and stored appropriately, In the experimental section, the basic rule is to record enough information to
e.g. in a ring binder). allow a reasonably competent scientist to repeat your work exactly. You must
●● Write clearly, taking care that indi- tread a line between the extremes of pedantic, irrelevant detail and the omission
vidual numerals cannot be confused. of information essential for proper interpretation – better perhaps to err on the
side of extra detail to begin with. An experienced worker can tell you which
subtle shifts in technique are important (e.g. batch numbers for an important
Electronic Laboratory Notebooks
chemical, or when a new stock solution is prepared). Many important scientific
(ELNs). – A number of commercial
advances have been made because of careful observation and record taking and
software products are available for tak-
because coincident data were recorded that did not seem of immediate value.
ing lab notes, e.g. Cambridge soft.
Make rough diagrams to show the arrangement of replicates, equipment, etc.
If forced to use loose paper to record data, make sure each sheet is dated and
taped to your lab book, collected in a ring binder, or attached with a treasury
tag. The same applies to traces, printouts and graphs.
The basic order of the primary record should mirror that of a research
report (see p. 601), including: the title and date, brief introduction, a compre-
hensive experimental section, the data and short conclusions.
Secondary record
You should make a secondary record concurrently or later in a bound book
and it ought to be neater, in both organisation and presentation. This book will
be used when discussing results with your supervisor, and when writing up a
report or thesis, and may be part of your course assessment. Writing a second,
neater version forces you to consider again details that might have been over-
looked in the primary record and provides a duplicate in case of loss or damage.
While these notes should retain the essential features of the primary record,

they should be more concise and the emphasis should move towards analysis
Analyse your data as soon as possi-
of the experiment. Don’t repeat the experimental section for a series of similar
ble – always analyse and think about
experiments; use devices such as ‘method as for Expt B4’. A photocopy may
data immediately after collection as
be sufficient if the method is derived from a text or article (check with your
this may influence your subsequent
supervisor). Outline the aims more carefully at the start and link the experiment
activities.
to others in a series (e.g. ‘Following the results of Expt D24, I decided to test
●● A graphical indication of what whether. . . ’). You should present data in an easily digested form, e.g. as tables
has happened can be particularly of means or as summary graphs. Use appropriate statistical tests (p. 491) to
valuable.
support your analysis of the results. Always analyse and think about data imme-
●● Carry out statistical analyses before
diately after collecting them as this may influence your subsequent activities.
moving on to the next experiment
because apparent differences Write down any conclusions: sometimes those which seem obvious at the time
among treatments may not turn out of doing the work are forgotten when the time comes to write up a report or
to be statistically significant when thesis. Likewise, ideas for further studies may prove valuable later. Even if your
tested. experiment appears to be a failure, suggestions as to the likely causes might
●● Write down any conclusions you prove useful.
make while analysing your data:
sometimes those which seem obvi-
ous at the time of doing the work are
Using communal records
forgotten when the time comes to If working with a research team, you may need to use their communal data-
write up a report or thesis. bases. These avoid duplication of effort and ensure uniformity in techniques.
●● Note ideas for further studies as they They may also form part of the legal safety requirements for lab work. They
occur to you – these may prove val-
might include:
uable later. Even if your experiment
appears to be a failure, suggestions ●● a shared notebook of common techniques (e.g. solutions or calibration
as to the likely causes might prove technique);
useful.
●● a set of simplified step-by-step instructions for use of equipment;
●● an alphabetical list of suppliers of equipment and consumables;
Safety Note Maintaining and
consulting communal lab records – ●● a list of chemicals required by the group and where they are stored;
these activities may form a part of the
●● the risk assessment sheets for dangerous procedures (p. 6);
safety requirements for working in a
laboratory. ●● a record of the use and disposal of chemicals and solvents.

Anon. Suggestions for Keeping Laboratory Notebooks. Currano, J. and Roth, D. (2013) Chemical information for
Available: http://otl.stanford.edu/inventors/resources/ Chemists. Royal Society of Chemistry, Cambridge.
labnotebooks.html Helmenstine, M.A. How to write a Lab Report. Available:
Last accessed 15/01/17. http://www.chemistry.about.com
[A Stanford University website which looks at the labo- Last accessed 01/07/09.
ratory notebook from a patent perspective] Zubrick, J.W. (2015) The Organic Chem Lab Survival
Beavon, J.R.G. Writing the Laboratory Notebook. Avail- Manual. A student’s guide to techniques, 10th edn. John
able: http://www.home.clara/rod.beavon/lab_book.htm Wiley & Sons Ltd, Chichester.

Resources
Eureka Lab Book. Available: www.eurekalabbook.com labfolder.com. Available: www.labfolder.com
Last accessed 05/01/16. Last accessed 05/01/16.
Electronic Lab Notebooks (ELNs): examples cambridge- labarchives.com. Available: www.labarchives.com
soft.com Available: www.cambridgesoft.com Last accessed 05/01/16.
Last accessed 05/01/16. Marsfield, N. (2007). Key Skills for scientists. Getting the
Hardbounds books: examples Mitchells Laboratory Note- message across. Section 2 Keeping a laboratory note-
books. Avaliable: www.laboratorynotebooks.co.uk book, Royal Society of Chemistry, Cambridge.
Study exercises
6.1 Design a primary data collection sheet for a lab of ten replicates for each of five different treat-
experiment. Imagine you wish to carry out a gravi- ments. Assume that you need to calculate means,
metric analysis (Box 22.1). what might you need to variances, etc., and then compare the results for
record on your data sheet? each treatment.
6.2 Design a secondary record table for the example
in 6.1. for the mass of precipitate formed in each
Answers to these study exercises are available at www.pearsoned.co.uk/practicalskills.

7 Project work
Research projects are an important component of the final-year syllabus for

most degree programmes in chemistry, while shorter projects may also be car-
ried out during courses in earlier years. Project work can be extremely reward-
ing, although it does present a number of challenges. The assessment of your
project is likely to contribute significantly to your degree grade, so all aspects
of this work should be approached in a thorough manner.
Deciding on a topic to study
Using the Internet as an information

Assuming you have a choice, this important decision should be researched
source for project work – since many
carefully. Make appointments to visit possible supervisors and ask them for
university departments and research
advice on topics that you find interesting. Use library texts and research papers
groups have homepages on the web,
to obtain further background information. Perhaps the most important criterion
searches using relevant key words may
is whether the topic will sustain your interest over the whole period of the
indicate where research in your area is
project. Other things to look for include:
currently being carried out Academics ●● Opportunities to learn new skills. Ideally, you should attempt to gain
usually respond positively to emailed experience and skills that you might be able to ‘sell’ to a potential employer.
questions about their area of expertise.
●● Ease of obtaining valid results. An ideal project provides a means to obtain
‘guaranteed’ data for your report, but also the chance to extend knowledge
by doing genuinely novel research.
Asking around – one of the best sources ●● Assistance. What help will be available to you during the project? A busy
of information about supervisors, labo- lab with many research students might provide a supportive environment
ratories and projects is past students. should your potential supervisor be too busy to meet you often; on the other
Some of the postgraduates in your hand, a smaller lab may provide the opportunity for more personal interac-
department may be products of your tion with your supervisor.
own system and they could provide an
alternative source of advice.
●● Impact. Your project may result in publishable data: discuss this with your
prospective supervisor.
Writing a proposal
You may be expected to submit a proposal for your project, especially where
it is expected that you should define the precise area of study yourself. The
structure for the proposal will probably be provided in the course handbook,
or as a form you must complete. Box 7.1 outlines some common features of
project proposals and what you should do to complete each part. The aim of
formulating a proposal is to ensure that:
●● the project has an appropriate theoretical background;
●● the objectives you have set are achievable;
●● the methods chosen are appropriate;
●● safety and ethical issues have been considered;
●● sufficient resources are available to complete the work, such as matching
students to available labs and supervisors;
●● you have set yourself a timetable with milestones on which your progress
can be judged;
●● you obtain feedback and suggestions about your plans.

Project work
Box 7.1 How to write a project proposal
Below are listed some common elements of project pro- (b) that you understand how to cite them properly
posals, with guidance regarding approach and content. (Chapter 41).
Not every part may be included, and the titles for sections
Research methods – this describes how you plan to
may differ – always follow the exact format specified for
carry out the investigation. Be quite specific, so the
your course.
committee can arrive at a valid judgement. Quote ref-
Contact details and suggested supervisor(s) – you erences for methods and techniques, where available.
may be asked whether you have discussed the project Consider use of appropriate controls, experimental
with the named supervisor(s). design and sampling procedures (Chapters 5 and 6).
Check that the results obtained will tell you what you
Proposed title – this should be relatively short, follow-
need to know to achieve your objectives and to test
ing the style used in research papers. It may change
your hypothesis.
for the final report.
Resources required – the samples, chemicals, instru-
Aims – a general statement of what you plan to
ments, etc. required to carry out your investigation.
achieve.
Listing these will require quite detailed consider-
Objectives – a listing of specific outcomes you expect ation of the experimental design or field area, as
to fulfil. Typically you will have several specific objec- relevant, and a thorough knowledge of methods,
tives that all fit within the overall aim. such as amounts or volumes of reagents used
(Chapters 8–12).
Brief description of the subject – this section might
have an alternative title like Summary, Background, Timetable/plan – a realistic breakdown of the work
Review of Subject Area or Statement of the Prob- required, with milestones leading to completion of
lem to be Addressed, it should contain a brief syn- the project (see Chapter 58 for advice on time man-
opsis of past work, a summary of current ideas and, agement). Always allow time for the unexpected.
if relevant, the hypothesis to be tested. In some
Statement or declaration in relation to safety – this
cases, it will consist of a mini-review of the subject
is to confirm that you have read and understood rel-
area and act as a template for the introduction to
evant issues, have completed relevant forms and
your report.
processes, and are in a position to proceed with the
Preliminary bibliography – this will ensure (a) that research. You may be required to attach copies of
you have read and understood relevant papers and COSHH/safety forms (Chapter 2).
Your proposal outline might be considered by your supervisor or by a

formal committee before approval.
Liaising with your supervisor(s) – this

is essential if your work is to proceed Planning your work
efficiently. Specific meetings may be
timetabled, e.g. to discuss a term’s
As with any lengthy exercise, planning is required to make the best use of the
progress, review your work plan or
time allocated (p. 533). This is true on a daily basis as well as over the entire
consider a draft introduction. Most
period of the project. It is especially important not to underestimate the time it
supervisors also have an ‘open-door’
will take to write and produce your thesis (see below). If you wish to benefit
policy, allowing you to air current prob-
from feedback given by your supervisor you should aim to have drafts in his/
lems or discuss your results. Prepare
her hands in good time. Since a large proportion of marks will be allocated to
well for all meetings; have a list of ques-
the report, you should not rush its production.
tions ready before the meeting; provide
If your department requires you to write an interim report, look on this
results in an easily digestible form (but
as a good opportuuity to clarify your thoughts and get some of the time-
take your lab notebook along) be clear
consuming preparative work out of the way. If not, you should set your own
about your future plans for work.
deadlines for producing drafts of the introduction, materials and methods
section, etc.

Project work
Choose project subject area

Key Point Project work can be very time consuming at times.
Try not to neglect other aspects of your course – make sure your
Read around subject
lecture notes are up to date and collect relevant supporting
information as you go along.
Create hypotheses and design

experiments Getting started
Fig. 7.1 is a flowchart illustrating how a project might proceed; at the start,
Consider safety don’t spend too long reading the literature and working out a lengthy pro-
aspects of work
including risk gramme of research. Get stuck in and do an experiment. There’s no substitute
assessment for ‘getting your hands dirty’ for stimulating new ideas;
(see Chapter 2)
●● even a ‘failed’ experiment will provide some useful information which may
allow you to create a new or modified hypothesis;
Start Start writing:
experimental • introduction ●● pilot experiments may point out deficiencies in experimental technique that
work; keep • materials and will need to be rectified;
detailed methods
records; • ideas for new
analyse and experiments
●● the experience will help you create a realistic plan of work.
graph data as and
you proceed discussion
• reference list
Designing experiments or sampling procedures
Design of experiments is covered in Chapter 5. Avoid being too ambitious at
the start of your work! It is generally best to work with a simple hypothesis and
Write final report
design your experiments or sampling around this. A small pilot experiment or
test sample will highlight potential stumbling blocks including resource limi-
Fig. 7.1 Flowchart showing a recommended
sequence of events in carrying out an tations, whether in materials or time or both.
undergraduate research project. Note the
potentially iterative nature of the process,
reflecting the scientific method outlined in Working in a laboratory environment
Chapter 5. During your time as a project student, you are effectively a guest in your super-
visor’s laboratory.
●● Be considerate – keep your ‘area’ tidy and offer to do your share of lab
duties such as calibrating the pH meter, replenishing stock solutions, distilled
water, cleaning used glassware, etc.
●● Use instruments carefully – they could be worth more than you’d think.
Careless use may invalidate calibration settings and ruin other people’s work
as well as your own.
●● Do your homework on techniques you intend to use – there’s less chance
of making costly mistakes if you have a good background understanding of
the methods you will be using.
●● Always seek advice if you are unsure of what you are doing.
Key Point It is essential that you follow all the safety rules
applying to the laboratory. Make sure you are acquainted with
all relevant procedures – normally there will be prominent warn-
ings about these. If in doubt, ask!

Project work
Keeping notes and analysing your results

Sources of further information for
project work: Tidy record keeping is often associated with good research, and you should
●● experimental design checklist follow the advice and hints given in Chapter 6. Try to keep copies of all files
(Box 5.1); relating to your project. As you obtain results, you should always calculate,
●● advice on recording results (p. 53); analyse and graph data as soon as you can (see Fig. 7.1). This can reveal aspects
●● describing and analysing numerical that may not be obvious in numerical or readout form. Don’t be worried by
data (p. 480); negative results – these can sometimes be as useful as positive results if they
●● checklist for presenting your final allow you to eliminate hypotheses – and don’t be dispirited if things do not
report (Box 70.1); work first time. Thomas Edison’s maxim ‘Genius is 1% inspiration and 99%
perspiration’ certainly applies to research work!
Writing the report

The structure of scientific reports is dealt with in Chapter 70. The following
advice concerns methods of accumulating relevant information.
Brushing up on IT skills – word proces-

Introduction This is a big piece of writing that can be very time-consuming.
Therefore, the more work you can do on it early on, the better. You should
sors and spreadsheets are extremely
allocate some time at the start for library work (without neglecting benchwork),
useful when producing a thesis. Chap-
ters 45 and 46 detail key features of
so that you can build up a database of references (p. 436). While photocopying
these programs. You might benefit from
can be expensive, you will find it valuable to have copies of key reviews and
attending courses on the relevant pro-
references handy when writing away from the library. Discuss proposals for
grams or studying manuals or texts so
content and structure with your supervisor to make sure your effort is relevant.
that you can use them more efficiently.
Leave space at the end for a section on aims and objectives. This is important
to orientate readers (including assessors), but you may prefer to finalise the
content after the results have been analysed!
Experimental You should note as many details as possible when doing the
experiment or making observations. Don’t rely on your memory or hope that
the information will still be available when you come to write up. Even if it is,
Using drawings or photographs – these
chasing these details might waste valuable time.
can provide valuable records of sam-
pling sites or experimental set-ups
and could be useful in your report.
Results Show your supervisor graphed and tabulated versions of your data
Plan ahead and do the relevant work at
promptly. These can easily be produced using a spreadsheet (p. 427), but
the time of carrying out your research
you should seek your supervisor’s advice on whether the design and print
rather than afterwards.
quality is appropriate to be included in your thesis. You may wish to access
a specialist graphics program to produce publishable-quality graphs and
charts: allow some time for learning its idiosyncrasies! If you are producing
a poster for assessment (Chapter 66), be sure to mock up the design well in
advance. Similarly, think ahead about your needs for any seminar or poster
you will present.
Using a word processor to record your
ideas – remember that you can note
down your thoughts and any other
Discussion Because this comes at the end of your thesis, and some parts
important information relevant to the
can only be written after you have all the results in place, the temptation is
Results and Discussion sections of your
to leave the discussion to last. This means that it can be rushed – not a good
project in a file that can then form the
idea because of the weight attached by assessors to your analysis of data and
basis of your first draft (p. 433); that
thoughts about future experiments. It will help greatly if you keep notes of
way, you won’t forget to include these
aims, conclusions and ideas for future work as you go along (Fig. 7.1). Another
points in your final report.
useful tip is to make notes of comparable data and conclusions from the liter-
ature as you read papers and reviews.

Project work
Acknowledgements Make a special place in your notebook for noting all

those who have helped you carry out the work, for use when writing this section
of the report.
References Because of the complex formats involved (p. 382), these can
be tricky to type. To save time, process them in batches as you go along –
bibliographic software (e.g. Endnote) can help with the organisation of refer-
ences (p. 382).
KEY POINT Make sure you are absolutely certain about the
deadline for submitting your report and try to submit a few days
before it. If you leave things until the last moment, you may find
access to printers, photocopiers and binding machines is difficult.

Ebel, H.F., Bliefert, C. and Russey, W.E. (2004) The Art of Overton, T., Johnson, S. and Scott, J. (2015) Study and
Scientific Writing: from Student Reports to Professional Communication Skills for the Chemical Sciences,
Publications in Chemistry and Related Fields, 2nd edn. 2nd edn., Oxford University Press, Oxford.
Wiley-VCH, Weinheim, Germany. Weyers, J. and McMillan, K. (2011) How to Write Disser-
Learn Chemistry. Royal Society of Chemistry. Available: www tations & Project Reports. 2nd edn., Pearson, Harlow.
.rsc.org/learn-chemistry/collections/Higher-Education/
he-resources/teaching-and-learning-chemistry
[Online Highr Education resources.]
Study exercises
Note: These exercises assume that you have started a write up the experimental section, analyse the data
research project, or are about to start one, as part of and draw up graphs as soon as you can. While you
your studies. may reject, or modify, some of these drafts at a
later stage, this approach will spread out the major-
7.1 Prepare a project plan. Make a formal plan for your
ity of the effort and allow time for critical thinking
research project, incorporating any milestones dic-
close to the final submission date.
tated by your department, such as interim reports
and final submission dates. Discuss your plan with 7.3 Devise a computer database for keeping details of
your supervisor and incorporate his or her com- your references. Keeping these records up to date
ments. Refer back to the plan frequently during will save you a lot of time when writing up. You
your project, to see how well you are meeting your will need to decide on an appropriate referencing
deadlines. format or find out about the one followed by your
department (see Chapter 41).
7.2 Resolve to write up your work as you go along. Each
time you complete an experiment or observation,

Fundamental laboratory techniques
8. Working with liquids 63
9. Basic laboratory procedures I 70
10. Basic laboratory procedures II 82
11. Principles of solution chemistry 102
12. pH and buffer solutions 113

8 Working with liquids
Measuring and dispensing liquids

Safety Note Using hazardous
liquids and solutions (flammable, cor- The equipment you should choose to measure out liquids depends upon the
rosive, toxic, etc.) – make sure the liq- volume to be dispensed, the accuracy required and the number of times the job
uid is properly contained and work out must be repeated (Table 8.1).
how to deal with any spillages before
you begin.
Table 8.1 Criteria for choosing a method for measuring out a liquid
Best volume Usefulness for repetitive

Reading any volumetric scale – make Method range Accuracy measurement
sure your eye is level with the bottom of
Pasteur pipette 1–5 mL Low Convenient
the liquid’s meniscus and take the read-
ing from this point. Conical flask/beaker 25–5000 mL Very low Convenient
Measuring cylinder 5–2000 mL Medium Convenient
Volumetric flask 5–2000 mL High Convenient
thumb and index Burette 1–100 mL High Convenient
finger provide
pressure on bulb Glass pipette 1–100 mL High Convenient
Mechanical pipettor 591000 mL High* Convenient
Syringe 0.5920 mL Medium** Convenient
Microsyringe 0.5950 mL High Convenient
Weighing Any (depends Very high Inconvenient
on accuracy of
balance
*If calibrated correctly and used properly (see p. 66).

**Accuracy depends on width of barrel: large volumes less accurate.
middle finger
at side of pipette
Conical flasks, beakers, measuring cylinders and volumetric flasks meas-
barrel supporting ure the volume of liquid contained in them, while burettes, pipettes, pipettors,
the pipette
syringes and microsyringes mostly measure the volume delivered from them:
Fig. 8.1 How to hold a Pasteur pipette. think about the requirements of the experiment.
Certain liquids may cause problems:
●● High-viscosity liquids are difficult to dispense: allow time for the liquid to
transfer.
●● Organic solvents may evaporate rapidly, making measurements inaccurate:
work quickly; seal containers quickly.
●● Solutions prone to frothing (e.g. surfactant solutions) are difficult to measure
and dispense: avoid forming bubbles; do not transfer quickly.
Safety Note Plastic, disposable Pasteur pipettes
Pasteur pipettes, ‘Pastettes’, may be Hold correctly during use (Fig. 8.1) – keep the pipette vertical, with the middle
used for aqueous solutions but NOT for finger gripping the barrel to support the pipette while the thumb and index
organic liquids. finger provide controlled pressure on the bulb, and squeeze gently to provide
individual drops.
To prevent liquid being sucked into the bulb and hence cross-contamination:
●● Ensure that the capacity of the bulb does not exceed that of the barrel.

Working with liquids
●● Do not remove the tip of the pipette from the liquid while drawing up the liq-
uid; the inrush of air may splash the liquid into the bulb. This is particularly
0 true when you lose patience trying to draw up viscous liquids.
1 10
●● Do not lay the pipette on its side during use.
2 9
Conversely, if volatile liquids such as dichloromethane (DCM), ethanol,
3 8 propanone (acetone) or diethylether (ether), for example, are to be dispensed,
4 7
the warmth of the glass pipette will cause the liquid to squirt from the pipette
25
without any pressure on the bulb. To prevent this, suck up the liquid several
5 6
times into the pipette so as to cool the glass and then dispense as normal.
6 5
Conical flasks and beakers
7 4
These have approximate graduations and should only be used for measuring
8 3 volumes of solutions/liquids where accuracy is unimportant.
9 2 Measuring cylinders and volumetric flasks
10 1 These must be used on a level surface (the laboratory bench) so that the scale is
horizontal; you should first fill with solution until just below the desired mark,
then fill slowly (e.g. using a Pasteur pipette) until the bottom of the meniscus
is level with the mark. Remember to allow time for the solution to run down
(a) (b) (c) the walls of the vessel and to bend down so that your eyes are level with the
graduation mark(s) and the meniscus.
Fig. 8.2 Glass pipettes: (a) graduated pip-
ette, reading from zero to shoulder; (b) Burettes
graduated pipette, reading from maxi- These must be mounted vertically in a clamp – don’t over-tighten the clamp
mum to tip, by gravity; (c) bulb (volumetric) (see p. 82) – or in a burette holder, on a stand. First ensure that the tap is closed
pipette, showing volume on bulb.
and, using a funnel, add a little of the solution to be dispensed, rinse the burette
and discard the washings through the tap: this is vital in titrations where a little
water in the burette will alter the concentration of the solution. Refill the burette
Glass pipettes – always check that the with solution, open the tap and allow the liquid to fill the barrel below the tap,
pipette is a ‘drain-down’ type. There then take a meniscus reading, noting the value in your notebook. Dispense
may be old ‘blow-out’ pipettes lurking the solution via the tap and measure the new meniscus reading. The volume
in the back of drawers. dispensed is the difference between the two readings.
Pipettes
There are various designs, including graduated and bulb (volumetric) pipettes
Using rubber bulb pipette fillers – infor- (Fig. 8.2). Take care to look at the volume scale before use: some graduated
mation on their use is given in pipettes empty from full volume to zero, others from zero to full volume; some
Chapter 24 (p. 195). scales refer to the shoulder of the tip, others to the tip by gravity. Never blow out
volumetric (bulb) pipettes, just touch the tip against the inside wall of the vessel.
Rinse out pipettes with a little of the solution to be delivered before com-
mencing the accurate measurement. To prevent cross-contamination, never
draw the solution into the pipette filler.
Key Point For safety reasons, it is no longer permissible to

Using a pipettor – check your mouth pipette – various aids (pipette fillers) are available, such
technique (precision) by dispens- as the rubber-bulb and Pi-Pump® (Fig. 8.3).
ing volumes of distilled water and
weighing on a balance, assuming
1 mg = 1 mL = 1 mm3. Fo r s m a l l Pipettors (autopipettors)
volumes, measure several aliquots There are two basic types:
together, e.g. 10 * 15 mL = 150 mg.
Aim for accuracy of {1%.
1. Air displacement pipettors. For routine work with dilute aqueous solu-
tions. One of the most widely used is the Gilson Pipetman® (Fig. 8.4). Box
8.1 gives details on its use.
64 Fundamental laboratory techniques

2. Positive displacement pipettors. For non-standard applications, includ-

evacuate ing dispensing viscous, dense or volatile liquids where an air displacement
bulb
pipettor might create aerosols leading to errors.
Air displacement and positive displacement pipettors may be:
fill
●● Fixed volume: capable of delivering a single factory-set volume.
pipette
●● Adjustable: where the volume delivered is determined by the operator
across a particular range of values.
●● Pre-set: movable between a limited number of values.
empty ●● Multichannel: able to deliver several replicate volumes at the same time.
pipette
Whichever type of these routine but expensive devices you use, you must
ensure that you understand the operating principles of the volume scale and the
(a) (b) method for changing the volume delivered – some pipettors are easily misread.
A pipettor must be fitted with the correct disposable tip before use and
Fig. 8.3 Pipette fillers: (a) rubber-bulb
each manufacturer produces different tips to fit particular models. Specialised
type; (b) Pi-Pump®.
tips are available for particular applications.
pushbutton If you accidentally draw liquid into the barrel, seek assistance from your
plunger demonstrator/supervisor since the barrel will need to be cleaned before further
tip ejector button
use (to prevent cross-contamination) and unskilled dismantling of the device
adjustment ring
will cause irreparable damage.
volume scale
(volumeter
Syringes
Syringes should be used by placing the tip of the needle into the solution and
slowly drawing the plunger up to the required point on the scale. Check the
barrel to make sure no air bubbles have been drawn up, and expel the solution
barrel
slowly, touching the needle tip on the side of the vessel to remove any adher-
ing solution. If there is air in the barrel, fill past the mark, invert the syringe
and push the plunger to the mark so that the air and a little of the solution are
expelled into a waste collection vessel. Then dispense the solution. The use of
syringes for dispensing air-sensitive reagents is described in Chapter 19.
tip ejector Microsyringes should always be cleaned before and after use by repeat-
edly drawing up and expelling pure solvent. The dead space in the needle can
occupy up to 4% of the nominal syringe volume. Some micro-syringes have
disposable tip a fine wire attached to the plunger, which fills the dead space. Never pull the
plunger out of the barrel.
Balances
Fig. 8.4 A pipettor – the Gilson Pipetman®.
These can be used to weigh accurately (p. 78) how much liquid you have dis-
pensed. Convert mass to volume using the equation:
Safety Note Using syringes – take
great care when handling syringe nee- Mass/density = volume
dles. They are very sharp and may be For example, a liquid (9.0 g) of density (1.2 g mL-1) = 7.5 mL. Densities
contaminated by chemicals. of common solvents and common chemicals can be found in Haynes (2015).
You will also need to know the liquid’s temperature, since density is temper-
ature dependent.
Using pipettors and syringes to deliver
small volumes – if you find that your
Holding and storing liquids
hard is ‘shaky’ when using these
devices, stabilise the body of the device Test tubes
with your second hard, making sure you
Both ‘normal’ and the much smaller ‘semi-micro’ test tubes are used for small-
do not touch the tip/needle.
scale reactions and tests, e.g. qualitative analysis (p. 179) or solvent selection
for recrystallisation (p. 130).

Box 8.1 Using a pipettor to deliver accurate, reproducible volumes of liquid
A pipettor can be used to dispense volumes with accu- measure several ‘squirts’ together, e.g. 20 ‘squirts’ of
racy and precision, by following this stepwise procedure: 5 mL = 100 mg. If the pipettor is inaccurate (p. 38) giv-
ing a biased result (e.g. delivering significantly more
1. Select a pipettor that operates over the appropriate or less than the volume set), you can make a tempo-
range. Most adjustable pipettors are accurate only rary correction by adjusting the volumeter scale down
over a particular working range and should not be or up accordingly (the volume delivered is more
used to deliver volumes below the manufacturer’s important than the value displayed on the volume-
specifications (minimum volume is usually 10%–20% ter), or have the pipettor recalibrated. If the pipettor
of maximum value). Do not attempt to set the vol- is imprecise (p. 38), delivering a variable amount of
ume above the maximum limit, or the pipettor may liquid each time, it may need to be serviced. After
be damaged. calibration, fit a clean (sterile) tip if necessary.
2. Set the volume to be delivered. In some pipettors, 5. Draw up the appropriate volume. Holding the
you ‘dial up’ the required volume. Types like the Gil- pipettor vertically, press down on the plunger/push-
son Pipetman® have a system where the scale (or ‘vol- button until a resistance (spring-loaded stop) is met.
umeter’) consists of three numbers, read from top to Then place the end of the tip in the liquid. Keeping
bottom of the barrel, and adjusted using the black your thumb on the plunger/push-button, release the
knurled adjustment ring (Fig. 8.4). This number gives pressure slowly and evenly: watch the liquid being
the first three digits of the volume scale and thus can drawn up into the tip, to confirm that no air bubbles
only be understood by establishing the maximum are present. Wait a second or so, to confirm that the
volume of the Pipetman®, as shown on the push-but- liquid has been taken up, then withdraw the end of
ton on the end of the plunger (Fig. 8.4). The following the tip from the liquid. Inexperienced users often
examples illustrate the principle for two common have problems caused by drawing up the liquid too
sizes of Pipetman®: quickly/carelessly. If you accidentally draw liquid into
P1000 Pipetman® P20 Pipetman®
the barrel, seek assistance from your demonstrator
(maximum volume 1000 mL) (maximum volume 20 mL) or supervisor as the barrel will need to be cleaned
if you dial up if you dial up before further use.
6. Make a quick visual check on the liquid in the

1 1
tip. Does the volume seem reasonable? (e.g. a
0 0 100 mL volume should occupy approximately half the
0 0 volume of a P200 tip). The liquid will remain in the tip,
without dripping, as long as the tip is fitted correctly
and the pipettor is not tilted too far from a vertical
the volume is set at 1000 mL the volume is set at 10.0 mL position.
7. Deliver the liquid. Place the end of the tip against

3. Fit a new disposable tip to the end of the bar- the wall of the vessel at a slight angle (10–15° from
rel. Make sure that it is the appropriate type for vertical) and press the plunger/push-button slowly
your pipettor and that it is correctly fitted. Press the and smoothly to the first (spring-loaded) stop. Wait
tip on firmly using a slight twisting motion – if not, a second or two, to allow any residual liquid to run
you will take up less than the set volume and liquid down the inside of the tip, then press again to the
will drip from the tip during use. Tips are often sup- final stop, dispensing any remaining liquid. Remove
plied in boxes, for ease of use: if sterility is important, from the vessel with the plunger/push-button still
make sure you use appropriate sterile technique at depressed.
all times. Never, ever, try to use a pipettor without its
disposable tip. 8. Eject the tip. Press the tip ejector button if present
(Fig. 8.4). If the tip is contaminated, eject directly into
4. Check your delivery. Confirm that the pipettor deliv- an appropriate container, e.g. a labelled container for
ers the correct volume by dispensing volumes of dis- hazardous solutions (p. 3). For repeat delivery, fit a
tilled water and weighing on a balance, assuming new tip if necessary and begin again at step 5 above.
1 mg = 1 mL = 1 mm3. The value should be within Always make sure that the tip is ejected before put-
1% of the selected volume. For small volumes, ting a pipettor on the bench.

Beakers
Beakers are used for general purposes, e.g. heating a non-volatile solvent while
the solute dissolves, ‘working up’ a reaction where liquid/solid products need
to be accessible for manipulation (stirring with a glass rod), or titrations using
electrodes where a wide opening is essential (see p. 305). The volume gradua-
tions on the side are inaccurate and should only be used where approximations
will suffice. The lip on the beaker is specifically designed to aid quantitative
transfer of solutions (see p. 75).
Volume accuracy – the experimen- Conical (Erlenmeyer) flasks

tal protocol may give an indication of These can be used for general purposes, but they have more specialist appli-
the equipment needed to measure the cations. The narrow mouth and sloping shoulders reduce losses on stirring/
volume of the liquid required. Terms swirling and evaporation and make them easier to seal. The absence of a lip
like 25.00 mL and 500.00 mL imply the does not favour quantitative transfer: useful in manual titrations (p. 194) and
use of a pipette or volumetric flask but recrystallisations (p. 133). Volume markings are approximate.
10 mL and 50 mL imply the use of a
measuring cylinder.
Bottles and vials
These can be used when the solution needs to be sealed for safety, storage
or to prevent evaporation or oxidation. They usually have a screwtop, plastic
Storing light-sensitive chemicals –
cap or stopper or a ground-glass stopper and come in various styles and sizes:
use a brown-glass vessel or wrap alu-
from 2.5 L glass bottles used for storing large volumes of solutions to small
minium foil around a clear vessel and
plastic-capped vials (5 mL) for saving small amounts of reaction products.
its stopper.
Seal the vessels in an appropriate manner, using a stopper, cap or sealing
film such as Parafilm® or Nescofilm®, bearing in mind the nature of the con-
tents – sealing film should only be used for water solutions since it dissolves in
some organic solvents and plasticisers may be extracted. Do not use corks, they
are not air-tight. Do not use rubber bungs to seal containers containing organic
solvents: they can swell even over a short period of time making removal very
difficult.
You should clearly label all stored solutions (see p. 76) including all
relevant hazard information.
Creating specialised apparatus

Glassware systems incorporating ground-glass connections, such as Quickfit®,
are essential for setting up combinations of standard glass components for reac-
tions, distillation, etc. In project work you may need to adapt standard forms
Table 8.2 Spectral cutoff values for glass of glassware for a special need. It may be necessary to contact a glassblowing
and plastics (l50 = wavelength at which service to make special items to order.
transmission of electromagnetic radiation
is reduced to 50%)
Choosing between glass and plastic containers
Material l50 (nm) Bear in mind the following points:
Routine glassware 340 ●● Reactivity. Plastic vessels often distort at relatively low temperature, may be
Pyrex glassware
®
292 inflammable, may dissolve in certain organic solvents and may be affected
Polycarbonate 396 by prolonged exposure to ultraviolet (UV) light.
Acrylic 342 ●● Opacity. Both glass and plastic absorb light in the UV range of the
Polyester 318 electromagnetic spectrum (Table 8.2). Quartz should be used where
this is important, e.g. in cells for UV spectrophotometry (see p. 218) or
Quartz 220
photochemistry.

●● Contamination. Some plasticisers may leach from vessels, especially

with some organic solvents, such as DCM. Glass may adsorb ions and
other molecules and then leach them into solutions, especially under acidic
or alkaline conditions. Pyrex® glass is stronger than ordinary soda glass
(rarely found except in specific items such as Pasteur pipettes and melting
point tubes, but check if you are not sure) and can withstand temperatures
up to 500°C.
●● Rigidity and resilience. Plastic vessels are not recommended where volume
is critical as they may distort through time. Glass vessels are more easily
broken than plastic.
●● Disposability. Plastic items may be cheap enough to make them disposable,
an advantage where there is a risk of chemical contamination.
Cleaning glass and plastic

Safety Note Special cleaning In quantitative analytical work, beware of contamination arising from prior
of glass – for an acid wash use dilute use of chemicals or inadequate rinsing following washing. A thorough rinse
acid, rinse with tap water (three times) with distilled or deionised water immediately before use will remove dust and
to remove the washing solution and other soluble deposits, but ensure that the rinsing solution is not left in the
then rinse thoroughly at least three vessel. For analyses on the ‘mg scale’ and below, you should clean glass and
times with distilled or deionised water. plastic containers by immersion in nitric acid (10% v/v) overnight and then
To remove acidic deposits, wash with rinsing with a large volume of distilled or deionised water. The clean vessels
KOH/ethanol solution followed by rins- should be stored either upside down or covered with Clingfilm®, to prevent
ing with deonised water. Glassware, dust contamination.
which must be exceptionally clean, For general work, ‘strong’ basic detergents (e.g. Decon® or Pyroneg®) are
should be washed in a chromic acid good for solubilising acidic deposits and an acid wash will remove remaining
bath, but this must only be made up basic residues. A rinse with ethanol or propanone (acetone) will remove many
and used under supervision. organic deposits.
Safety with glass

Many minor accidents in the laboratory are due to lack of care with glassware.
You should follow these general precautions:
●● Wear eye protection at all times.
WRONG ●● Don’t use chipped or cracked glassware and examine the equipment for ‘star’
cracks – it may break under very slight strains and should be disposed of in
the broken glassware bin. All laboratories will have a waste bin dedicated to
broken glass. Never put broken glass into other bins.
●● If heating glassware, use a ‘soft’ Bunsen flame (half-open air vent) or ‘wave’
the flame around the heating point – this avoids creating a hot spot where
cracks may start. Always use special heat-resistant gloves or rubber ‘fingers’
(see p. 92) when handling hot glassware.
●● When clamping glassware (see p. 82) ensure that the clamp has a cork, rub-
ber or plastic ‘cushion’ in the jaws to prevent breakages. There must be no
metal – glass contact and you must not over-tighten the clamp.
RIGHT
●● Take care when attaching rubber or plastic tubing to glass tubes, condensers,
etc., and inserting thermometers and glass tubes into screwcap adapters (see
p. 100). Always hold the tube and the ‘hole’ close together (Fig. 8.5) and
Fig. 8.5 Handling glass tubing. wear thick gloves where appropriate.

●● Don’t force bungs too firmly into bottles (see p. 67), they can be difficult
to remove. If you need a tight seal, use a screwtop bottle, with a rubber or
plastic seal, Parafilm® or ground-glass jointware, such as Quickfit®.
●● Never carry large bottles (7 1 L) by their necks – carry them in a bottle
basket.
●● Dispose of broken glass thoroughly and with great care – use disposable
paper towels, tongs or dust-pan and brush and thick gloves. Always put
pieces of broken glass in the correct bin.
Text reference
Haynes, W.M. (ed.) (2015) CRC Handbook of Chemistry
and Physics, 96th edn. CRC Press, Boca Raton.

Beram, J.A. (2014) Laboratory Manual for Principles
of General Chemistry, 10th edn. John Wiley & Sons,
Chichester.
Study exercises
8.1 Decide on the appropriate methods and equip- 8.2 Write a protocol for calibrating and using a pipet-
ment for the following procedures: tor. After reading this chapter, prepare a street-
wise protocol explaining how to use a pipettor to
(a) Preparing one litre of ethanol at approximately
deliver a specific volume, say 500 mL (e.g. using a
70% v/v in water for use as a general-purpose
Gilson Pipetman® or an alternative if your depart-
reagent.
ment does not use this type). Ask another student
(b) Adding 25.00 mL of concentrated hydrochloric
to evaluate your protocol and provide you with
acid to water (100 mL).
feedback – either by reading through your protocol
(c) Adding 10 mL of propanone to a volumetric
or by trying it out with a pipettor as part of a class
flask (100.00 mL) to produce an aqueous solu-
exercise (check with a member of staff before you
tion calibration standard for chromatography.
attempt this in a laboratory).
(d) Titrating an unknown solution of sodium
hydroxide (approximate concentration 0.1 M)
with sulphuric acid (0.5 M).

9 Basic laboratory procedures I
Using chemicals
Finding out about chemicals – The
Merck Index (O’Neil et al., 2013) and the
CRC Handbook of Chemistry and Phys-
Safety aspects
ics (Haynes, 2015) are useful sources on In practical classes, the person in charge has the responsibility to inform you
the chemical and physical properties of of any hazards associated with the use of chemicals. In project work, your
elements and compounds, including first duty when using an unfamiliar chemical is to find out about its proper-
melting and boiling points, solubility ties, especially those relating to safety. For routine practical procedures, a risk
and toxicity, etc. Manufacturers’ cat- assessment may have been carried out by a member of staff and the relevant
alogues now include hazard data and safety information may be included in the practical schedule: an example is
disposal procedures. shown in Table 9.1. In project work, your first duty when using an unfamiliar
chemical is to find out about its properties, especially those relating to safety.
Your department must provide the relevant information to allow you to do this.
Table 9.1 Representative risk assess-
If your supervisor has filled out the form, read it carefully before signing.
ment information for a practical exercise
in organic chemistry: the synthesis of
N-phenylethanamide (acetanilide) Key Point Before you use any chemical you MUST find out
whether safety precautions need to be taken, and complete
Substance Hazards/comments
the appropriate forms confirming that you appreciate the risks
Aminobenzene Toxic, harmful by involved.
skin absorption. Wear
gloves, dispense in
fume cupboard Essential safety points when handling chemicals are:
Ethanoic anhydride Corrosive, flamma- ●● Treat all chemicals as potentially dangerous.
ble, toxic, reacts with
water. ●● Wear a lab coat, with the buttons fastened, at all times.
Wear gloves, dispense ●● Wear eye protection at all times.
in fume cupboard
●● Make sure you know where safety devices such as eye bath, fire extinguisher,
first aid kit are kept before you start work in the lab.
Using chemicals responsibly – be con-
●● Wear gloves for toxic, irritant or corrosive chemicals and carry out proce-
siderate to others: always return store-
dures with them in the fume cupboard.
room chemicals promptly to the correct
place. Report when supplies are getting ●● Use appropriate aids such as spatulae, pipette fillers, Pasteur pipettes, etc. to
low to the person responsible for look- minimise risk of contact.
ing after the store. If you empty an aspi-
●● Label all solutions, samples, etc. appropriately (see p. 78).
rator or wash bottle, fill it up from the
appropriate source. ●● Extinguish all naked flames when working with flammable substances.
●● Never drink, eat or smoke where chemicals are being handled.
Molar solutions – you will find that ●● Report all spillages and clean them up appropriately.
chemists talk about ‘0.1 Molar solutions’ ●● Dispose of chemicals in the correct manner.
or you may see ‘0.1 M’ as a concentra-
tion written on flasks or in books and Selection
journals. The term ‘Molar’ (abbreviated
to M) means number of moles per litre.
Chemicals are supplied in varying degrees of purity and this is always stated
Hence an aqueous solution of hydro-
on the manufacturer’s containers. Suppliers differ in the names given to the
chloric acid (0.1 M) has a concentration
grades and there is no conformity in purity standards. Very pure chemicals
of 0.1 mol L-1 equivalent to 3.65 g of
cost more, often very much more, and should only be used when the situation
hydrogen chloride per litre of solution.
demands. If you need to order a chemical, your department will have a defined
procedure for doing this.

Preparing solutions
Solutions are usually prepared with respect to their molar concentrations (e.g.
mol L-1, mol dm -3 or mol m -3) or mass concentrations (e.g. g L-1 or kg m -3):
both can be regarded as an amount (usually mass) per unit volume, in accord-
ance with the relationship:
amount
concentration = [9.1]
volume
Percentage concentration – you may The most important aspect of eqn [9.1] is to recognise clearly the units
find that the concentration of a solution involved, and to prepare the solution accordingly: for molar concentrations
is expressed in percentage terms. Thus you will need the relative molecular mass of the chemical, so that you can
sodium carbonate (5% w/v) – the sym- calculate the mass of substance required. Further advice on concentrations and
bol w indicates weight of solute and v interconversion of units is given on p. 102.
is the volume of solution. The resulting In general, there are two levels of accuracy required for the preparation
solution is a general-purpose dilute of solutions:
aqueous solution of sodium carbonate 1. General-purpose solutions – solutions of chemicals used in qualitative and
prepared from sodium carbonate (5 g) preparative procedures (p. 72) when the concentration of the chemical
made up to 100 mL in water and used need not be known to more than one or two decimal places. For example:
for neutralising acid.
(a) solutions used in extraction and washing processes, e.g. hydrochlo-
ric acid (0.1 mol L-1), sodium hydroxide (2 M), sodium carbonate
(5% w/v);
(b) solutions of chemicals used in preparative experiments where the
techniques of purification – distillation, recrystallisation, filtration,
etc. – introduce intrinsic losses of substances that make accuracy to
The levels of accuracy of solution prepa- any greater level meaningless.
ration required are usually indicated in
the protocol or by the nature of the 2. Analytical solutions – solutions used in quantitative analytical procedures
experiment. – Look for phrases such (p. 74) when the concentration needs to be known to an accuracy of four
as ‘accurately weighed’ which means decimal places (e.g. 0.0001 mol L-1). For example in:
to four decimal places on an analyti-
cal balance, together with quantitative (a) volumetric procedures (titrations) and gravimetric analysis, where the
transfer. Volumes quoted as 250.00 mL, concentrations of standard solutions of reagents and compounds to be
100.00 mL, 25.00 mL imply the use of analysed need to be accurately known;
volumetric flasks and pipettes. Volumes (b) spectroscopy, e.g. quantitative UV and visible spectroscopy, atomic
given in mL imply the use of pipettors absorption spectroscopy and flame photometry;
or syringes.
(c) electrochemical measurements: pH titrations, conductance measure-
ments and polarography;
(d) chromatographic methods.
The procedures for weighing and the glassware used in the preparation of
solutions differ according to the level of accuracy required.
Preparation of general-purpose solutions
Box 9.1 shows the steps involved in making up general-purpose aqueous
solutions.
The concentration you require is likely to be defined by the protocol you
are following and the grade of chemical and supplier may also be specified. To
avoid waste, think carefully about the volume of solution you require, though
it is always advisable to err on the high side because you may spill some, make

Box 9.1 How to make up an aqueous solution of known concentration from a solid chemical
1. Find out or decide the concentration of chemical (e) T h e r e f o r e , y o u n e e d t o m a k e u p

required and the degree of purity necessary. 0.2 * 286.14 = 57.2 g up to 100 mL of solution
using distilled water.
2. Decide on the volume of solution required.
3. Find out the relative molecular mass of the chem- 5. Weigh out the required mass of chemical to an appro-
ical (Mr). This is the sum of the atomic (elemental) priate accuracy. If the mass is too small to weigh with
masses of the component element(s) and can usually the desired degree of accuracy, consider the follow-
be found on the container. If the chemical is hydrated, ing options:
i.e. has water molecules associated with it, these must
be included when calculating the mass required. (a) Make up a greater volume of solution.
(b) Make up a more concentrated solution, which
4. Work out the mass of chemical that will give you can be diluted at a later stage.
the concentration desired in the volume required,
bearing in mind the quoted percentage purity of the (c) Weigh the mass first, and calculate what volume to
chemical. make the solution up to afterwards using eqn [9.1].
Example 1: Suppose your procedure requires you to

6. Add the chemical to a beaker or conical flask and then
prepare 250 mL of 0.1 mol L-1 sodium chloride solution.
add a little less water than the final volume required.
(a) Begin by expressing all volumes in the same units, If some of the chemical sticks to the weighing recep-
either millilitres or litres (e.g. 250 mL as 0.25 L). tacle, use some of the water to wash it off. For accu-
rate solutions, see p. 74 for accurate weighing and
(b) Calculate the number of moles required from eqn
quantitative transfer.
[9.1]: 0.1 = amount (mol) , 0.25.
By rearrangement, the required number of moles 7. Stir and, if necessary, heat the solution to ensure all
is thus 0.1 * 0.25 = 0.025 mol. the chemical dissolves. You can determine visually
when this has happened by observing the disappear-
(c) Convert from mol to g by multiplying by the relative
ance of the crystals or powder. Allow the solution to
molecular mass (Mr for NaCl = 58.44 g mol-1).
cool, if heated.
(d) T h e r e f o r e , y o u n e e d t o m a k e u p
0.025 * 58.44 = 1.46 g up to 250 mL of solution, 8. Make up the solution to the desired volume. If the
using distilled water. concentration needs to be accurate, use a volumetric
flask (see p. 74 for accurate weighing and quantitative
Example 2: Suppose you are required to make up 100 transfer); if a high degree of accuracy is not required,
mL of sodium carbonate (2 M). use a measuring cylinder.
(a) Convert 2 M into mol L-1; concentration (a) Pour the solution from the beaker into the meas-
required = 2 mol L-1. uring vessel using a funnel to avoid spillage,
using water to rinse out the vessel.
(b) Express all volumes in the same units: therefore
100 mL = 0.1 L. (b) Make up the volume so that the meniscus comes
up to the appropriate measurement line. If accu-
(c) Calculate the number of moles required from racy is not a major concern, the graduation marks
eqn [9.1]: 2 = amount (mol) , 0.1. By rear- on the beaker or conical flask may be used to
rangement, the required number of moles is thus establish the approximate volume.
2 * 0.1 = 0.2 mol.
(d) Convert from mol to g by multiplying by the Mr
9. Transfer the solution to a reagent bottle or conical
but note from the container that the compound
flask and label the vessel clearly, including hazard
is Na2CO3.10H2O. Therefore the Mr required
information, where appropriate. Do not use water in
must include the water of crystallisation and
this final transfer since you will alter the concentra-
Mr = 286.14 g mol-1.
tion of the solution by dilution.
a mistake when dispensing or need to repeat part of the experiment. Choose

one of the standard volumes for vessels, as this will make measuring out easier.
Use distilled or deionised water to make up aqueous solutions and stir with
a clean Pyrex® glass rod or magnetic stirrer bar (‘flea’) until all the chemical

is dissolved. Magnetic stirrers are a convenient means of stirring solutions but

precautions should be taken to prevent losses by splashing. Add the flea to the
empty beaker or conical flask, add the chemical and then some water. Place the
vessel centrally on the stirrer plate, switch on the stirrer and gradually increase
the speed of stirring. When the crystals or powder have dissolved, switch off
the stirrer and remove the flea with a magnet. Take care not to contaminate
your solution when you do this and rinse the flea into the solution with distilled
water. In general, it is convenient to use glass rods with beakers – ease of access
for stirring – and magnetic fleas with conical flasks – lower losses through
splashing – but often it is a matter of your preference and laboratory skills.
‘Obstinate’ solutions may require heating, but only do this if you know that
the chemical will not be damaged at the temperature used. Use a stirrer-heater
to keep the solution mixed as you heat it. Allow the solution to cool to room
temperature before you finalise its volume.
Preparation of analytical solutions

The key features in the preparation of solutions for analytical purposes are:
●● Make sure that you have the most accurate available knowledge of masses
of the chemicals used.
●● Ensure that you have the most accurate available knowledge of the volumes
of solutions used.
To achieve these features exact techniques of weighing and solution trans-
fer must be used and the procedure is illustrated in the following example.
‘Prepare a standard solution (250.00 mL) of ammonium ferrous sulphate
(approximately 0.1 M), which is to be used to determine the concentration
A standard solution is one in which
of a solution of potassium permanganate by titration’.
the solute is weighed out to an accuracy
of 4 decimal places and is made up in a You must be aware of the following embedded information:
volumetric flask.
●● This is a quantitative experiment, therefore requiring an analytical solution
A stock solution is one from which to be prepared.
dilutions are made.
●● You must use a 250.00 mL volumetric flask, which you should note is cal-
ibrated at 20 °C.
●● You must weigh accurately, to four decimal places, the mass of the chemical.
●● It is almost impossible to weigh the exact mass of chemical for a specific
concentration. For example, the mass of (NH4)2FeSO4.6H2O required to
prepare 250.00 mL of 0.1 M solution is 9.8035 g and you cannot weigh out
this exact mass. However, you can weigh out a known mass to four decimal
places accurately. From this you can then calculate the exact concentration
of the chemical in solution, since you will know both mass and volume to a
high degree of accuracy.
Box 9.2 shows the method for the preparation of the standard solution. The
main practical point is that you must not lose, by splashing or failure to transfer
by inadequate rinsing, any of the solution being prepared in the beaker and you
must transfer all of the solution, by repeated rinsing, into the volumetric flask.
Therefore it is good practice to use only a glass rod to stir the solution gently
to dissolve the solid and to use the glass rod, as shown in Fig. 9.1, to pour the
solution into the filter funnel. This technique, with practice, prevents losses of
solution down the side of the beaker via the spout; rinsing with water can be
achieved by use of a wash bottle squirted directly into the beaker.

Box 9.2 H
ow to make up an aqueous solution of known concentration from a solid chemical
for use in quantitative analysis
Example: Suppose you are to prepare a standard solu- chemical in the beaker: 11.9726 - 2.1564 = 9.8162 g
tion (250.00 mL) of ammonium ferrous sulphate (approx- of (NH4)2FeSO4.6H2O.
imately 0.1 M), which is to be used to determine the
concentration of a solution of potassium permanganate. 7. Add deionised or distilled water (about 100 mL) to
the beaker and stir the mixture gently with a clean
1. This is a quantitative experiment so the ammonium Pyrex® glass rod until all the solid has dissolved. Do
ferrous sulphate must be of the highest purity avail- not splash or spill any of this solution or you can-
able to you. not calculate its concentration. Remove the glass rod
from the solution, rinsing it with a little distilled water
2. Work out the mass of chemical that will give you the into the solution.
concentration desired in the volume required.
8. Clamp a clean volumetric flask for support (see p. 82)
(a) Convert 1.0 M into mol L-1; concentration and place a clean, dry filter funnel in the top supported
required = 1.0 mol L-1. by a ring. Carefully pour the solution into the volumet-
(b) Express all volumes in the same units: therefore ric flask, ensuring no spillage of solution by using the
250.00 mL = 0.25 L. technique illustrated in Fig. 9.1 and pouring slowly so
(c) Calculate the number of moles required from that no air-lock is formed and no solution runs down the
eqn [9.1]: 1.0 = amount (mol) , 0.25. By rear- side of the beaker. When the addition is complete, do
rangement, the required number of moles is thus not move the beaker from its position over the funnel.
1.0 * 0.25 = 0.025 mol. 9. Rinse the inside of the beaker several times with
(d) Convert from mol to g by multiplying by the Mr, a distilled water wash bottle to transfer all of the
but note from the container that the compound solution into the volumetric flask, paying particular
is (NH4)2FeSO4.6H2O. Therefore the Mr required attention to the ‘spout’ and glass rod. Then place the
must include the water of crystallisation and beaker aside and lift the funnel from the flask while
Mr = 392.14 g mol-1. rinsing it with distilled water. You have now achieved
(e) Therefore, you need to weigh out a quantitative transfer. Swirl the liquid in the flask to
0.025 * 392.14 = 9.8035 g of (NH4)2FeSO4.6H2O. prevent density gradients.
10. Make the solution up to the mark using distilled

3. Place a clean, dry weighing boat or appropriately water, stopper the flask and mix thoroughly by gen-
sized sample tube onto a simple two-decimal-place tle inversion (10 times) of the flask while holding the
balance (see p. 78) and zero (tare) the balance and stopper in place.
weigh about 9.80 g of the chemical. You now have a solution (250.00 mL), which con-
tains (NH4)2FeSO4.6H2O (9.8162 g).
4. Carefully transfer the sample tube plus chemical to a The concentration of this solution is expressed as:
four-decimal-place analytical balance (see p. 79) and
record the accurate mass: say 11.9726 g. 250.00 mL of the solution contains 9.8162 g of
(NH4)2FeSO4.6H2O
5. Remove the sample and container from the balance
and tip the contents into a clean, dry beaker (400 mL), Therefore:
ensuring that there is no spillage outside the beaker. 1000.00 mL of solution contains
Do not attempt to wash out the sample tube with
(4 * 9.8162) = 39.2648 g of (NH4)2FeSO4.6H2O
water.
The concentration of the solution is
6. Immediately reweigh the sample tube on the ana-
lytical balance: say 2.1564 g. This is the mass of 39.2648 g L-1 = 39.2648 , 392.14 =
the container together with a few crystals of the
chemical which have remained in the container. 0.1001 mol L-1 = 0.1001 M
However, you now know exactly the mass of the
You should not use a flea to stir a solution in the preparation of a standard
solution, since this introduces more washing steps – washing the flea and the
‘flea extractor’ – and you still need to use the glass rod for quantitative transfer.

Procedure required with analytical solutions prepared from liquids

Many experiments in analytical chemistry, such as chromatography and spec-
troscopy, require the preparation of a standard solution of a liquid organic
compound. Therefore you must know accurately the mass of the liquid. The com-
pound can be dispensed by the methods described in Chapter 8, provided that
the pipette, syringe, etc., is accurate, and thus the mass = volume * density,
bearing in mind the temperature factor.
Alternatively you can use a weighing bottle as shown in Fig. 9.2. The
liquid is placed in the bottle, weighed accurately and then the approximate
amount required is added to the volumetric flask containing some solvent. The
volumetric flask is stoppered immediately, the weighing bottle reweighed and
the weight of liquid dispensed is calculated. The volumetric flask is then made
up to the mark and stoppered. You now know the concentration of the standard
solution to four decimal places.
Fig. 9.1 Pouring a solution using a glass rod. Preparing dilutions

Making a single dilution
teat
In analytical work, you may need to dilute a standard solution to give a
particular mass concentration or molar concentration. Use the following
Pasteur pipette
procedure.
1. Transfer an appropriate volume of standard solution to a volumetric flask,
using appropriate equipment (Table 8.1).
screwcap adapter 2. Make up to the calibration mark with solvent – add the last few drops
from a Pasteur pipette until the bottom of the meniscus is level with the
calibration mark.
3. Mix thoroughly, either by repeated inversion (holding the stopper firmly)
or by prolonged stirring, using a magnetic stirrer. Make sure that you add
5mL Quickfit® conical flask
the magnetic flea after the volume adjustment step.
For general-purpose work using dilute aqueous solutions where the higher
Fig. 9.2 A weighing bottle.
degree of accuracy is not required, it may be acceptable to substitute conical
flasks, beakers or test tubes for volumetric flasks and use measuring cylinders for
volume measurements. In such cases you would calculate the volumes of ‘stock’
Making a dilution – use the relation-
solutions (usually ‘bench’ reagents) and diluent required, with the assumption
ship [C1]V1 = [C2]V2 to determine vol-
that the final volume is determined by the individual volumes of stock solution
ume or concentration (see p. 103).
and diluent used. Thus a two-fold dilution would be prepared by using one vol-
ume of stock solution and one volume of diluent. The dilution factor is obtained
from the initial concentration of the stock solution and the final concentration
Using the correct volumes for dilu- of the diluted solution. The dilution factor can be used to calculate the volumes
tions – it is important to distinguish and stock and diluent required in a particular instance. For example, suppose that
between the volumes of the various you wanted to prepare 100 mL of a solution of NaOH at 0.1 mol L-1. Using the
liquids: a one-in-ten dilution is obtained bench reagent, commonly containing 2.0 mol L-1 (2.0 M), the dilution factor is
using one volume of stock solution plus 0.1 , 2.0 = 0.05 = 1/20 (a 20-fold dilution). Therefore the amount of stock
nine volumes of diluent (1 + 9 = 10). solution required is 1/20th of 100 mL = 5 mL and the amount of diluent needed
is 19/20th of 100 mL = 95 mL.
Preparing a dilution series

Dilution series are used in a wide range of procedures including the preparation of
standard curves for the calibration of analytical instruments (p. 445). A variety of
different approaches can be used but the most common is a linear dilution series.

In a linear dilution series, the concentrations are separated by an equal

amount, e.g. a series containing cadmium at 0, 0.2, 0.4, 0.6, 0.8, 1.0 mmol L-1
might be used to prepare a calibration curve for atomic absorption spectros-
copy (p. 342) when assaying polluted soil samples. Use [C1]V1 = [C2]V2 to
calculate the volume of standard solution for each member of the series and
pipette or syringe the calculated volume into an appropriately sized volumet-
ric flask as described above. Remember to label clearly each diluted solution
as you prepare it, since it is easy to get confused. The process is outlined in
Box 9.3.
Key Point Make all the dilutions from the working stock solu-
tion to the required solution. Do not make a solution of lower
dilution from one already prepared: if you have made an error
in the first dilution, it will be repeated for the second dilution.
Mixing solutions and suspensions

Solutions must be thoroughly mixed before measuring out volumes for the next
dilution. Use a fresh measuring vessel for each dilution to avoid contamination,
or wash your vessel thoroughly between dilutions. Clearly label the vessel con-
taining each dilution when it is made: it is easy to get confused! When deciding
on the volumes required, allow for the aliquot removed when making up the
next member in the series. Remember to discard any excess from the last in the
series if volumes are critical.
Various devices may be used, including:
●● Magnetic stirrers and fleas. Magnetic fleas come in a range of shapes and
sizes, and some stirrers have integral heaters. During use, stirrer speed may
increase as the instrument warms up.
●● Orbital shakers and shaking water baths. To provide controlled mixing
at a particular temperature, e.g. for long-term incubation.
●● Bottle rollers. For liquid–liquid extraction work, ensuring gentle, contin-
uous mixing.
Safety Note Using a vortex mixer
with open or capped test tubes – do not
●● Vortex mixers. For vigorous mixing of small volumes of solution, e.g.
vortex too vigorously or liquid will spill
when preparing a dilution series in test tubes. Take care when adjusting the
from the top of the tube, creating a con-
mixing speed – if the setting is too low, the test tube will vibrate rather than
tamination risk,
creating a vortex, giving inadequate mixing. If the setting is too high, the test
tube may slip from your hand.
Storing chemicals and solutions
Sealing flasks – think carefully about
the sealing system for a flask to be Chemicals which decompose easily (labile chemicals) may be stored in a fridge
stored at low temperature. Cooling will or freezer. Take special care when using chemicals which have been stored at
reduce the volume of vapour (includ- low temperature: the container and its contents must be allowed to warm up to
ing air) in the flask and create a partial room temperature before use, otherwise water will condense onto the chemical.
vacuum. Rubber bungs can be irre- This may render accurate weighing impossible and you may ruin the chemical.
movable after cooling (another good Chemicals and solutions to be stored at low temperatures must be in
reason for never using them) and even stoppered or sealed vessels. Do not store aqueous solutions below 0 °C since
ground-glass joints can seize up. Plastic freezing can occur and, with the resulting expansion of the volume, the vessel
stoppers, screwtops or lightly greased may crack. Solutions containing flammable solvents should only be stored in
glass stoppers, as appropriate, are specialised ‘spark-proof’ fridges: consult your laboratory instructor.
recommended. You must be aware of the particular problems of storing solutions in flasks
with ground-glass joints. If you are using aqueous solutions you should lightly

Box 9.3 How to make up a linear dilution series for use in quantitative analysis
The experimental protocol states: ‘Prepare a standard 5. Transfer the standard solution (2.00 mL) to the volu-
solution (250.00 mL; 0.01 M) of cadmium ions using cad- metric flask (100.00 mL) and make up to the mark with
mium nitrate and use this solution to produce a linear distilled water.
dilution series of solutions (100.00 mL) of accurately
known concentrations of approximately 0.0, 0.2, 0.4, 0.6, 6. Repeat the calculation for each of the other diluted
0.8 and 1.0 mmol L-1. solutions as required, but note that a short cut
is possible in this case: since you require stock
1. Calculate the amount of cadmium nitrate required for solution (2.00 mL) for the diluted solution of con-
the standard solution: cadmium nitrate is supplied as the centration 0.2 mmol L-1, you will need 4.00 mL for the
tetrahydrate Cd(NO3)2 . 4H2O; Mr = 308.47 g mol-1. 0.4 mmol L-1 solution, etc. Use pure distilled water
Therefore, using eqn [9.1], you should calculate for the solution of concentration 0.0 mol L-1.
250.00 mL of a 0.01 M solution of cadmium ions requires
0.01 * 0.25 = 0.0025 mol = 0.0025 * 308.47 7. Calculate the exact concentrations of the diluted
= 0.7712 g of Cd(NO3)2 . 4H2O. Remember: this solu- solutions using [C1]V1 = [C2]V2, since the concen-
tion will contain 0.0025 moles of ‘cadmium nitrate’ tration of the standard solution is most unlikely to
or 0.0025 moles of cadmium ions and 0.005 moles of be exactly 0.0100 mol L-1 (see Box 9.2). For example,
nitrate ions. if the concentration of the standard solution [C1] is
actually 0.009 87 mol L-1 = 9.87 * 10-3 mol L-1, then
2. Make the standard solution by the quantitative for the dilute solution of approximate concentration
method described in Box 9.2 and calculate its con- 0.2 mmol L-1, the actual concentration [C2] is:
centration to four decimal places.
3. Calculate the volume of solution required for [C2] = {V1 * [C1]} , V2

the dilution series using [C1]V1 = [C2]V2, where 0.2 mL * 9.87 * 10-3 mol L-1
V1 = volume (mL) of standard solution required; =
100.00 mL
C1 = concentration of standard solution;
V2 = volume of diluted solution (100.00 mL) and = 2 * 9.87 * 10-5 mol L-1 = 0.1974 mmol L-1.
C2 = concentration of diluted solution.
Note: It is much simpler to measure out whole-number
4. Express the concentrations in the same units, the volumes (2.00 mL, 4.00 mL, etc.) using a pipette and pro-
most convenient in this case being mol L-1. Therefore, duce diluted solutions of accurately known concentra-
C1 = 0.01 M = 1 * 10-2 mol L-1 and for the diluted tions (but not necessarily whole numbers) rather than to
solution of concentration 0.2 mmol L-1, C2 = 0.2 * try to produce whole-number concentrations by measur-
10 - 3 mol L-1. By rearrangement ing out non-whole-number volumes.
V1 = {[C2]V2} , [C1]
0.2 * 10-3 mol L-1 * 100.00 mL

= = 2.00 mL
1 * 10-2 mol L-1
grease the joint and stopper with petroleum jelly, since the water will not dis-
solve the grease as it is poured from the flask. The stopper can be removed
easily and the solution will be uncontaminated.
Conversely, if you are using solutions made up from organic solvents, you
Greasing joints – if you are using should not grease the joints since the organic solvent will dissolve the grease as
solutions of NaOH or KOH you must you pour it from the flask and contaminate the solution. Moreover, you should
grease the ground-glass joint and stop- not allow the solution to come in contact with the ungreased joints, since the
per since the surfaces are attacked by solvent will evaporate and leave the solute to ‘weld’ the stopper to the flask.
strong alkalis. Fill the flask with solution, using a filter funnel with the stem of the funnel
positioned well below the joint. See p. 98 for use of ground-glass jointware.

Key Point Always label all stored chemicals clearly with the
Weighing – never weigh anything
following information: the name (if a solution, state solute(s)
directly onto a balance’s pan: you may
and concentration(s)), plus any relevant hazard warning infor-
contaminate it for others. Use an appro-
mation, the date made up, and your name.
priate weighing container such as a
weighing boat, sample tube, weighing
paper, conical flask, beaker. Balances and weighing
Electronic single-pan balances with digital readouts are now favoured over
mechanical types and are common in most laboratories. There are essentially
‘Weighing paper’ – it is common prac-
two types of balance:
tice to put a piece of paper onto the pan
of general-purpose balances. The mass 1. General purpose balances which weigh to the nearest 0.01 g with a capac-
of the paper is then ‘tared off’ before ity of about 300 g. Chemicals may be dispensed for weighing, into a suit-
the weighing container is placed on able weighing container, directly onto these balances.
the balance pan. The paper protects the
2. Analytical four-figure balances for quantitative work, which weigh to the
balance pan from corrosion by spillages
nearest 0.0001 g (0.1 mg) and have a maximum capacity of about 100 g.
and also allows you to discard easily
Chemicals must not be transferred onto the balance at any time and ana-
any material spilt without affecting the
lytical balances must only be used for weighing by difference.
weighing.
Both types are illustrated in Fig. 9.3 and you should familiarise yourself
with their operation before use.
General-purpose balances
The most useful feature of this type of balance is the electronic zero facility
(self-taring), which means the mass of the weighing container can be subtracted
automatically before weighing chemicals.
To operate a standard self-taring balance:
1. Check that it is level, using the adjustable feet to centre the bubble in the
spirit level (usually at the back of the machine). For relatively accurate
work or when using in a fume cupboard, make sure that the draught shield
(a) is in place.
2. Ensure that the balance is switched on: the display should be lit.
3. Place an empty weighing container (see p. 79) centrally on the balance pan
and allow the reading to stabilise. If the object is larger than the pan, take care
that no part rests on the body of the balance or the draught shield as this will
invalidate the reading. Press the tare bar to bring the reading to zero.
4. Place the chemical or object carefully in the weighing vessel:
(a) Solid chemicals should be dispensed with a suitably sized clean
spatula.
(b) Non-volatile liquids should be dispensed using a Pasteur pipette but
take the weighing container off the balance pan before dispensing;
then reweigh the liquid plus container. Repeat until the desired weight
is obtained.
5. Allow the reading to stabilise and make a note of the reading.
6. If you have added excess chemical, take great care when removing it.
(b) Remove the container from the balance, remove the solid (with a spatula)
or liquid (with a Pasteur pipette) and reweigh.
Fig. 9.3 Single-pan balance: (a) general
purpose, two decimal places; (b) analytical, 7. If you need to clean any deposit accidentally left on or around the balance,
four decimal places. switch off the balance.

Take care not to exceed the limits for the balance: while most have devices
to protect against overloading, you may damage the mechanism.
Analytical balances
These are delicate precision instruments and as such are likely to be
found away from the open laboratory in draught-free conditions on a
v ibration-dampened surface. Analytical balances are maintained to the
highest specifications and should need no adjustments on your part, such as
levelling and zero adjustment. The key points for using an analytical balance
are summarised below:
●● No chemicals must be transferred within the weighing compartment of the
balance.
Using a balance – It is poor experi-
●● If it has a ‘locking’ function, the balance pan must always be ‘locked’ when
mental technique to use large contain-
placing and removing objects onto and from the balance pan.
ers to weigh out small masses: you
are attempting to measure small dif- ●● The doors of the balance must always be closed when taking measurements.
ferences between large numbers. For
The procedure for weighing a solid chemical for the preparation of an analytical
small masses, use a small weighing
standard solution is shown in Box 9.4.
container (Fig. 9.4).
Weighing containers
These come in various materials, shapes and sizes: from glass weighing boats
to beakers and even special glazed paper. The weighing container to be used
depends on several factors:
●● The amount of chemical to be weighed.
●● The properties of the chemical to be weighed: is it solid, liquid, volatile,
corrosive, deliquescent, hygroscopic?
●● How and into what type of vessel it is to be transferred.
(a)
●● The accuracy to which it is to be weighed.
Some of the common types of weighing container are shown in Fig. 9.4.
For analytical procedures, only weighing boats, weighing bottles or glass
or plastic sample tubes should be used. Weighing boats are used to transfer a
solid directly into a volumetric flask via the neck of the weighing boat: this
procedure is recommended when the chemical is known to be totally soluble in
the solvent and allows you to omit the solution preparation stage in a beaker or
conical flask (see Box 9.2). You must ensure that the neck of the weighing boat
(b) will fit well inside the ground-glass joint of the neck of the volumetric flask so
that all the chemical can be washed down the sides of the volumetric flask and
does not stick to the ground-glass joint during the quantitative transfer.
For general-purpose work, weighings can be made directly into pre-
weighed or ‘tared’ conical flasks or beakers, again to avoid a transfer stage.
Much more common is the use of disposable plastic weighing dishes of the
appropriate size. The edges of these dishes can be squeezed together to form
a ‘funnel’ to prevent losses when transferring the solid. Remember that plastic
disposable dishes may dissolve in organic solvents such as propanone (ace-
(c)
tone), toluene, etc., and should not be used for low-melting organic solids or
liquids. Watch- and clock-glasses should be avoided if you wish to transfer the
Fig. 9.4 Weighing containers: (a) plastic solid into narrow-necked vessels such as conical flasks or sample tubes since
weighing dishes; (b) weighing bottles; (c) it is very difficult to direct the solid into the narrow opening of the vessel from
weighing boat. the large ‘flat’ surface of the watch- or clock-glass. In such cases, or when large

Box 9.4 H
ow to weigh out a sample of a solid for use in
1. Place the clean, dry weighing boat or sample tube on a general-

purpose two-decimal-place balance and zero the balance.
2. Weigh out the calculated amount of chemical within the accuracy of

the balance.
3. Check the zero reading on the analytical balance by pressing the bar/
button with the balance doors closed.
4. Relock the balance pan by pressing the bar/button.

5. Carefully transfer the weighing boat or sample tube to the balance
pan of the analytical balance (for very accurate work use tweezers
or fine tongs since the sweat from your fingers will contribute to the
weight recorded) and close the balance door.
6. Release the balance pan by pressing the bar/button, allow the balance
to stabilise and record the weight of the chemical and container. If the
last decimal place ‘cycles’ between two or three numbers, determine
the mid-point of the ‘cycle’ and record this value as the weight.
7. Lock the balance pan by pressing the bar/button, remove the sample
container and transfer the solid to your volumetric flask, beaker or
conical flask by pouring, but do not wet the weighing boat or sample
tube with solvent.
8. Replace the weighing container on the analytical balance pan, close

the balance door and weigh the container. Again decide on the mid-
point weight if the last decimal place ‘cycles’ and record this value
as the weight of the ‘empty’ weighing container.
9. Lock the balance pan by pressing the bar/button and remove the
weighing container from the balance.
Fig. 9.5 Transferring a solid using glazed
paper. 10. Subtract the weight of the ‘empty’ weighing container from that of
the weighing container plus sample and you now know the mass
of chemical, to an accuracy of four decimal places, which has been
transferred into your volumetric flask, beaker or conical flask.
amounts of solid are to be transferred, it is advisable to use a wide-necked filter

folded glazed paper
funnel called a ‘powder funnel’.
solid In many preparative experiments, which are carried out on a small scale
(involving 1 g to 10 g of solids), the most useful weighing container is spe-
rolled glazed paper
cial glazed paper, provided that the chemicals do not react with the paper.
A creased square of glazed paper is ‘tared’ on the balance pan and the solid
weighed out directly onto it. The chemical can then be allowed to flow down
the crease into the vessel (Fig. 9.5). Furthermore, when attempting to transfer
small amounts of solid in vessels with narrow-bore ground-glass joints (see
flask p. 98) it is important not to allow the solid to contact the joint, because the joint
will not seal correctly. Use a filter funnel or roll a piece of glazed paper into
a funnel, insert the stem of the paper funnel to below the joint and then run in
the solid from the creased weighing paper (Fig. 9.6). Paper used in this manner
Fig. 9.6 Transferring a solid to a narrow-necked is much cheaper than proprietary weighing dishes and is a useful method of
flask. recycling out-of-date manufacturers’ catalogues!

Text references
Haynes, W.M. (ed.) (2015) CRC Handbook of Chemistry and Biologicals, 15th edn. Merck & Co. Inc., White-
and Physics, 96th edn. CRC Press, Boca Raton. house Station.
O’Neil, M.J., Smith, A. and Henkelmann, P.E. (2013) The
Merck Index: An Encyclopaedia of Chemicals, Drugs

Anon. SIRI MSDS Index. Available: http://www.hazard Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch-
.com/msds ell, A.R. (1991) Vogel’s Textbook of Practical Organic
Last accessed 05/03/16. Chemistry, 5th edn. Longman, Harlow, Essex.
[Online access to Materials Safety Data Sheets (MSDS) Mendham, J., Denney, R.C., Barnes, J.D. and Thomas,
for manufacturers’ chemicals] M.J.K. (2000) Vogel’s Textbook of Quantitative Chemi-
cal Analysis, 6th edn. Prentice Hall, Harlow, Essex.
Study exercises
9.1 Practise the calculations involved in preparing spe- (a) If you added sodium chloride solution
cific volumes of aqueous solutions of known con- (1.00 mL; 0.4 M) to a volumetric flask
centrations. What mass of substance would be (10.00 mL) and made up to the mark with
required to prepare each of the following (answer water, what would be the concentration of
in each case to four decimal places): the diluted solution?
(b) Calculate the volume a solution of copper (II)
(a) 250.00 mL of sodium chloride (0.05 M) from
sulphate (0.1 M) required to produce 500.00
NaCl?
mL of 0.02 M solution.
(b) 100.00 mL of potassium iodate (0.02 M) from
(c) What would be the concentration of a solution
KIO3?
of Fe2+ ions if 10.00 mL is diluted to 250.00 mL
(c) 50.00 mL of sodium thiosulphate (0.05 M) from
to give a concentration of 0.001 M?
Na2S2O3.5H2O?
(d) You are provided with a solution of
(d) 250.00 mL of copper ions (0.1 M) from
KMnO4(0.02 M); to what volume must 5.00 mL
CuSO4.5H2O?
of this solution be diluted to give a concentra-
(e) 100.00 mL of potassium ions (0.05 M) from
tion of 0.001 M?
K2SO4?
9.2 Practise the calculations involved in dilu-

tions (answer in each case to four decimal places).

10 Basic laboratory procedures II
Clamps and support stands

When carrying out experiments it is vital that all apparatus is held in place
securely during the procedure. It is essential that you know how to assemble
supporting and securing equipment to the highest possible standards of safety.
The most common types of supporting and securing equipment are shown in
Fig. 10.1.
Support stands
These are also known as ‘retort stands’ and comprise an aluminium or steel rod
screwed into a heavy metal base. Always check that the base sits level and that
the rod is tightened fully in place.
support stand
Clamp holders
These are also described as ‘clamp bosses’ or ‘bosses’. Make sure that the
locking screws move freely and are not distorted. When you attach the clamp
holder to the support stand, tighten the screw firmly and ensure that the open
‘slot’ to be used for the clamp is pointing upwards (Fig. 10.2).
Clamps
three-finger clamp clamp holder General-purpose clamps are used for securing glassware – therefore make sure
that the inner surfaces of the clamp ‘jaws’ and the ‘fingers’ are covered with
cork or rubber to provide a cushion for the glass: there must be no metal to
glass contact in case you overtighten the clamp and crush the glass. Tighten the
clamp firmly and ensure that the clamped glassware does not move.
Conical flasks should be clamped at the neck and ground-glass joint-
ware should be clamped at the joint – this usually has the greatest thickness
clamp metal ring
of glass.
Key Point Take particular care when using parallel-sided

separatory funnels and chromatography columns (Fig. 10.3),
where clamping in the middle of the funnel can be the same as
squeezing the middle of a large-diameter glass tube. Clamp at
the ground-glass joint using a ‘well-cushioned’ clamp.
burette clamp
Fig. 10.1 Clamps and supporting equipment. In most clamps, only one of the jaws moves when turning the screw. When
you use the clamp in a horizontal position, make sure that the movable jaw is
at the top (see p. 147).
Burette clamps are specially designed to hold the burette vertically. Springs
hold the burette at two points about 5 cm apart – again check for the presence
of a rubber or plastic ‘cushion’ at the points of contact – to prevent slipping and
lateral movement. Since the burette clamp slides down the rod of the support
stand, then provided the support stand is vertical, the burette will be vertical.
Support rings
Using support rings – make sure that
These metal rings come in various diameters to support filter funnels and sep-
the tap on the separating funnel will go
aratory funnels. Often these support rings are coated in plastic to provide the
through the ring.
cushion between metal and glass.
If your support ring is metal, you can make a ‘cushion’ by finding a piece
of thin-walled rubber tubing of the same bore as the metal of the ring, cutting
it to a length equivalent to the circumference of the ring and then cutting down

Basic laboratory procedures II
the length of the rubber tubing. You can then slide the tubing around the ring
to provide the ‘cushion’.
Safety Note When using clamps, support rings and support

RIGHT WRONG stands make sure that the clamped/supported apparatus is always in
position above the base of the support stand (Fig. 10.4) to prevent the
movable jaw on TOP stand toppling over.
Cork rings
These are used to hold round-bottom flasks on flat surfaces while manipula-
fixed jaw underneath to
WRONG tions are being carried out. Since they are light in weight they can be used to
support, e.g. a condenser
hold round-bottom flasks on a general-purpose balance.
Fig. 10.2 Right and wrong ways of using
support stands, clamp holders and clamps. Filtration
Filtration is the physical separation of a solid from a liquid and is a process
encountered in experimental procedures such as gravimetric analysis (p. 184),
recrystallisation (p. 128) and solvent drying (p. 95). In principle, the mixture of
Gravity filtration to obtain a solid – the solid and liquid is passed through a porous material, filter paper or sintered
there are a few occasions when these glass, and the solid is trapped on the porous material while the liquid passes
rules do not apply, e.g. when the solid through.
is to be decomposed by heat and the The type of filtration equipment you select for use depends upon which of
filter paper destroyed, a process known the two components, the solid or the liquid, you are trying to isolate. In general:
as ‘ashing’. Some gravimetric analyses,
such as the determination of sulphate ●● If you wish to isolate the liquid – use gravity filtration.
as barium sulphate, require retention ●● If you wish to isolate the solid – use suction (vacuum) filtration
of the precipitate from gravity filtration
since BaSO4 is too fine to be collected Gravity filtration
on a vacuum filtration system. BaSO4 In gravity filtration you need to pass the liquid through the porous material
is thermally stable up to 600 °C, so the and retain all the unwanted solid in the filter. In general, the best material to
filter paper can be burned away. use is a filter paper of the appropriate porosity to trap all the solid particles
and with the greatest surface area to allow the liquid to pass through quickly.
The apparatus required for gravity filtration is shown in Fig. 10.5. The filter
funnels are usually made of glass, but if organic solvents are not involved in
the filtration, plastic funnels can be used. Glass filter funnels with the pipe
cut off are known as ‘stemless’ filter funnels and have a specific use in hot
filtration (p. 134).
The key to successful gravity filtration is the fluted filter paper. A fluted
filter paper decreases the area of contact between the filter paper and the funnel,
thus allowing rapid filtration. If you use ‘traditional’ cone-folded filter paper,
note that all sides of the paper are touching the sides of the funnel and on half
the filter paper the liquid has to pass through three thicknesses of paper, all
of which slow the rate of filtration. Slow filtration can lead to disaster in hot
filtration during recrystallisation (p. 136).
Since filter funnels and filter papers come in different sizes, choose a filter
paper of diameter just less than twice the diameter of the funnel. When fluted,
the filter paper will be just below the rim of the funnel. There are many ways
to fold (flute) a filter paper, but one of the simplest is shown in Box 10.1.
To filter the mixture, swirl the suspension of the solid in the liquid so that
there is a fairly even distribution of solid in the liquid, and then pour the mix-
RIGHT
ture into the filter cone, making sure that you do not pour any of the mixture
Fig. 10.3 Clamping a parallel-sided funnel. outside the filter paper otherwise you will need to repeat the filtration, and do

Box 10.1 How to flute a filter paper for gravity filtration
1. Fold the filter paper in half (a), open it out then fold into quarters
(b), open it out and then fold into eighths (c) and (d), all in the same
direction.
2. Turn the paper over and fold each sector in half (e); you are creating
sixteenths but with each fold in the opposite direction to (a), (b), (c)
and (d).
3. Finally, fold the paper into a cone (g) ensuring that all the folds are
RIGHT WRONG sharp and that the base of the cone comes to a sharp point.
Fig. 10.4 Correct use of support stands. 4. The flutes ensure that the filter paper has minimum contact with the
filter funnel and the sharp point ensures that the liquid flows rapidly
out of the cone and out of the funnel.
5. If your cone point is blunt it will cover the stem of the funnel and so
all the liquid must pass through this part of the filter paper, slowing
the filtration.
ring clamp
supported
on stand
(a) (b) (c)
(f) (e) (d)
Fig. 10.5 Gravity filtration.

(g)
not overfill the filter cone. Transfer all the mixture in this way and finally wash
Filtering solids – if you allow the solid
the last bit of solid and liquid into the filter cone with a small amount of filtered
to settle you will have difficulty pouring
solution and then a little pure solvent.
the thick slurry of solid and liquid from
the bottom of the beaker or conical flask Suction filtration
into the filter cone and you will need to This technique is used for the isolation of a solid from a suspension of a solid
use more filtrate to complete the filtra- in a liquid and relies on producing a partial vacuum in the receiving flask. The
tion effectively. essential components of a suction filtration system are:
●● A ceramic funnel containing a flat perforated plate: there are two types based
on size and shape called Büchner funnels or Hirsch funnels. When you are
filtering, the perforated plate is covered by a filter paper.
●● A receiver flask with a side arm for attachment of the vacuum source. Büch-
ner flasks are conical flasks made from thickened glass, and Hirsch tubes

(also known as side-arm boiling tubes or test tubes depending upon size) are
Filtering hot mixtures – never attempt
capable of withstanding weak vacuum, e.g. a water pump.
to use suction filtration on a hot mix-
ture of solid and liquid. As the liquid ●● A flexible seal between the ceramic funnel and the receiving vessel: a Büch-
filters through into the vacuum it may ner collar or filter seal.
boil under the reduced pressure and be
●● A source of vacuum, usually a water pump (water aspirator) which is con-
drawn into the pressure tubing. If the
nected to the receiving flask by thick walled rubber tubing (pressure tubing).
liquid is saturated with the solid, the
Usually there will be a trap between the water pump and the receiving flask.
evaporating liquid will deposit the solid
in the holes of the perforated plate and The various types of these components are shown in Fig. 10.6: typical appa-
block them. ratus is shown in Fig. 10.7 and the general procedure for suction filtration is
described in Box 10.2.
Filtering charcoal – never attempt to

remove finely divided charcoal, used
in decolorisation during recrystallisa-
tion or as a catalyst support, by suction
filtration. It is a very fine powder and
will always leak into the filtrate. Filter
off charcoal by gravity filtration.
Büchner funnel Hirsch funnel
Büchner collar
side-arm
Büchner flask boiling tube
or test tube
Fig. 10.6 Equipment for suction filtration.
thick-walled tubing
flat filter paper vacuum release tap
to vacuum
Büchner
collar
trap
clamp
thick-walled filter flask

(Büchner flask)
Fig. 10.7 Suction filtration using a Büchner funnel.

Box 10.2 Isolation of a solid by suction filtration
1. Select the appropriate size of apparatus based on the need to pull it off quickly if something goes wrong
amount of solid you expect to isolate and the volume (see 1B and 5). The filter paper will be pulled down
of liquid to be collected in the receiver flask. Consider onto the perforated plate by the vacuum.
the following points:
9. Turn on the tap to the water pump to the maximum
A. There is no point in using a large Büchner funnel water flow. If you do not do this, the water pump is
for a small amount of solid since you will collect a not working at its maximum efficiency and the vac-
layer of solid ‘one molecule thick’ and be unable uum created in your filtration system may cause
to scrape it from the filter paper cleanly. If there is water to be sucked into a trap, or receiving flask, from
too much solid for the size of the funnel, you will the water pump. This is called ‘suck-back’.
have to repeat the filtration with a second set of
apparatus or the solid may not suck dry quickly. 10. Swirl the mixture to be filtered and then slowly pour
it into the Büchner or Hirsch funnel at such a rate
B. If you use a side-arm boiling tube to try to col- so that the filtration is rapid. Note that the rate of
lect 100 mL of liquid, you will overfill the tube filtration may slow as the ‘cake’ of solid on the filter
and liquid: (i) will flow into the pressure tubing, becomes thicker.
contaminating it for your fellow students; (ii) may
fill the intermediate trap, if there is one, and you 11. To transfer the last of the solid/liquid from its beaker
will need to dismantle and clean it; (iii) may be or conical flask into the funnel use a little of the filtrate
sucked into the water pump causing corrosion in the receiving flask. Release the vacuum by opening
and loss of performance. the tap on the trap or pull off the vacuum tubing, but
do not turn off the tap on the water pump (there is a
2. Clean and dry all the apparatus to be used.
possibility of ‘suck-back’ (see 9 above)). Dismantle the
3. Clamp the receiving vessel to a support stand: pres- apparatus, pour a little of the filtrate into the beaker or
sure tubing is heavy and even large Büchner flasks conical flask, reassemble the apparatus and continue
will fall over: do not think that a test tube rack will the filtration. Repeat until all the material has been fil-
hold a side-arm boiling tube safely. tered. Use the filtrate to wash down any of the solid
sticking to the sides of the funnel onto the filter ‘cake’
4. Place the correct-sized Büchner collar in the neck of
– it will not dry quickly on the sides of the funnel.
the receiving flask: it should sit well into the neck
and fit the funnel to form a good seal. 12. Release the vacuum, by pulling the vacuum tubing
5. Place the funnel into the collar/seal: note that the from the flask or opening the tap on the trap and
funnel has a ‘point’ at the bottom of the stem. Make turn down the water pressure on the water pump.
sure that this ‘point’ is as far away as possible from Transfer the filtrate to a clean beaker or conical flask.
the vacuum attachment side arm of the receiver Add a little pure, ice-cold solvent to the filter cake and
flask, since the filtering liquid runs off this ‘point’ and reconnect the vacuum to provide gentle suction. This
if the point is near the vacuum inlet, the liquid may will wash the solid. Turn up the vacuum to maximum
be drawn into the side arm and then into the trap or and suck air through the solid to dry it as much as
water pump (see 1B above). possible. If ‘cracks’ appear in the filter ‘cake’, close
them by pressing gently with a clean spatula and
6. Select a filter paper, which fits exactly over the per- repeat until no more filtrate appears to be sucked out.
foration in the base of the funnel. The filter paper
should not fold or crease up the sides of the funnel 13. When drying is complete, release the vacuum, turn
because the solid will be sucked round the edge of off the water tap and remove the filter funnel from
the paper into the receiver flask. If the paper does not the apparatus. The solid is best removed as a com-
fit exactly, trim to size with scissors. plete ‘cake’ by lifting the edge of the filter paper with
spatula, inverting the funnel over a watch-glass or
7. Place the paper into the funnel and wet it with a few
clock-glass and the cake should fall out. Peel the filter
drops of liquid – the same liquid which is to be used
from the top of the ‘cake’, break up the ‘cake’ using a
in the filtration.
spatula and dry it appropriately.
8. Switch on the tap for the water pump to provide
14. Evaporate the filtrate to half volume (see p. 161) and
gentle suction. If your system has a trap, don’t forget
cool to obtain a second crop of crystals.
to close the tap on the trap and connect the rubber
tubing to the side arm of the receiver. Do not force 15. Wash out and clean all the apparatus and dispose of
the rubber tubing too far onto the side arm – you may the liquid filtrate safely.

sintered-glass vacuum take-off

Sintered-glass funnels – never use a filter disk side-arm
sintered-glass funnel to remove finely
divided charcoal. It is always absorbed
into the pores of the sinter and almost
impossible to remove easily. sintered-glass
filter disk
standard
sinter for taper joint
gravimetric sinters for
analysis filtration
Fig. 10.8 Typical sintered-glass funnels.
Removing charcoal from a cold solution

Instead of a filter paper on a porous plate, sintered-glass can be used as the
by suction filtration – cover the filter
porous material for filtration. Sintered-glass funnels come in various types and
paper with a ‘filter aid’ such as Celite®.
sizes (Fig. 10.8) with different porosity (size of holes) of the sintered glass. Sin-
This absorbs the fine particles.
tered glass is also used in crucibles (Fig. 10.8) in gravimetric analysis (p. 184).
When sintered-glass funnels are used instead of Büchner or Hirsch funnels
the solid is collected directly on the sintered glass: no filter paper is used. As a
result cleaning sintered-glass funnels is a major problem if the solid has been
Safety Note Never use gas burn-
drawn into the pores of the sintered-glass. Therefore, if you are to use a sintered
ers to heat flammable chemicals in
glass funnel, check with your instructor on the appropriate method for cleaning
open containers, in particular solvents,
the funnel, before and after use, so that your product will not be contaminated.
which are usually used in relatively
On the other hand, if particle sizes are large enough to prevent this problem,
large volumes.
sintered-glass funnels are very effective in suction filtration.
Heating
In the laboratory you will be required to heat chemicals in dissolution of a
Safety Point Before you start solid, promotion of reaction (reflux), distillation of pure compounds and mix-
heating a liquid or solution you must tures, extraction, coagulation of precipitates, drying solid compounds, etc. Your
always take one of the following choice of heat source depends upon several factors:
‘anti-bumping’ precautions:
●● Add one or two ‘boiling stones’ or
●● First and foremost, the flammability and volatility of the chemical and solvent.
‘anti-bumping granules’; these can ●● The operation to be carried out, e.g. simple preparation of a solution, reflux
be filtered off later in the process. or distillation.
●● Add a Pyrex® glass rod to the beaker
or conical flask. The rod must be ●● The temperature required for the process.
longer than the container so that it
can be removed and rinsed before ●● The amount of chemical or solvent to be heated.
the solution is used further. Bumping
●● Add a ‘boiling stick’: these are thin
pieces of wood sold as ‘wooden Before you attempt to heat any liquid or solution you must take precautions to
applicators’, but you must be sure prevent ‘bumping’. This is when the liquid suddenly boils without any warning
that nothing will be extracted from and results in hot liquid and vapour shooting uncontrollably out of the con-
the wood into your solution. tainer. ‘Bumping’ can occur during simple heating in a test tube, conical flask
●● Stir the liquid or solution with a or beaker or in more complex situations such as reflux and distillation. It is
magnetic ‘flea’ (see p. 90) which necessary to provide a point in the liquid or solution where vaporisation of the
should be removed before further liquid can occur in a controlled manner.
processing of the chemicals.
●● Stir the liquid or solution with a Burners
mechanical stirrer (see p. 157). Gas burners come in two common forms: large burners called Bunsen burn-
●● Use an air or inert gas (nitrogen)
ers and small burners known as microburners (Fig. 10.9). Bunsen burners are
capillary bleed during vacuum distil-
lation (see p. 149).
commonly used for heating aqueous solutions in flat-bottomed Pyrex® vessels
supported on a tripod and wire gauze (Fig. 10.10) but for many other heating

Safety Note Using a micro-

burner – do not place the microburner hot blue flame
when air inlet open
under the flask and leave it there: you
will have no control over the rate of
luminous wavy flame
heating. when air inlet closed
movable collar
movable collar
air inlet
Heating a test tube – you must ‘wave’ air inlet
the test tube in the flame to prevent
localised heating, which will cause
the liquid to ‘bump’ out of the tube.
Also use an appropriate anti-bumping
device and make sure that the tube is (a) (b)
not pointing at anyone.
Fig. 10.9 (a) Bunsen burner; (b) microburner.
Key Point If you forget to take ‘anti-bumping’ precautions

you MUST allow the liquid to cool down before you attempt the
‘anti-bumping’ process, otherwise the liquid will boil uncontrol-
lably out of the flask.
applications Bunsen burners do not provide adequate control. Microburners

may be used for direct heating of round-bottom or pear-shaped glassware in
small-scale operations where good control of the heating rate is required, such
as distillation or determination of melting point (see p. 125). When using a
microburner for heating make sure that you do not create a ‘hot spot’, which
may result in uneven heating of the liquid and ‘bumping’, by ‘waving’ the
flame around the flask starting just below the level of the liquid and working
down to the bottom of the flask and back again.
Microburners are also useful for heating boiling tubes and test tubes, seal-
ing the ends of melting point tubes (see p. 125), making micropipettes for chro-
matography (see p. 261) and bending the ends of Pasteur pipettes for special
purposes (see p. 151).
To use a burner, first make sure that the gas piping is attached securely
Fig. 10.10 Heating an aqueous solution to both the burner and the gas tap, is of the correct type of rubber tubing, and
using a Bunsen burner.
is not damaged. Close the movable collar and light the gas with a gas igniter:
do not use a splint or a piece of burning paper in case you cause a fire in the
Tubing for gas burners – this must be waste bin when disposing of the paper. Open the collar to produce a hot blue
of medium wall thickness so that it does flame and adjust the gas flow to give the required size of flame. If your work is
not kink or compress easily. With thin- interrupted, close the collar a little to produce a luminous flame. Finally, when
walled tubing, you could accidentally the operations are complete, turn off the gas and do not pick up the burner by
lean on the tubing, cutting off the gas the barrel, or put it into a cupboard, until it has cooled.
supply, which extinguishes the burner. The advantages of the burner are that the heat source can be removed
When you release the constriction, gas instantly from the apparatus and that the flame is visible: in contrast you often
will then flow into the laboratory. cannot distinguish between a cold and hot metal surface. The disadvantages
are those of an open flame.
Steam baths and water baths
Safety Note Before lighting a gas
burner make sure that nobody is using
These are heat sources, which have a maximum temperature of 100 °C; they are
flammable solvents in the laboratory.
safe to use even with most flammable chemicals and solvents and they differ
only in the way the steam is produced.

Water baths are the more common of the two, and the steam is gen-
Safety Note Take particular care erated by heating water with an electric element – just like a kettle. The
when you turn off the gas supply to a element may have a thermostatic control, which can control the temperature
Bunsen burner or microburner. Usually of the water to some extent. Most water baths have a constant level device
the gas tap is situated at the back of the on the side of the bath, which supplies water to the bath to a fixed level
laboratory bench, so make sure you do above the heating element and prevents the water bath from boiling dry.
not reach over the flame or knock over Water baths are ‘single hole’ or ‘multiple hole’ types, and the holes are
your apparatus. covered by concentric metal or plastic rings, which allows you to vary the
size of the hole.
When you are to use a water bath, make sure that water is flowing into
Do not confuse a water bath, which
and out of the bath via the constant level device – check that water is flowing
is a general-purpose heating device,
from the pipe into the drain. Turn on the controller to the level you require – the
with a constant temperature bath,
power can be turned down once the bath is boiling.
which is a precision device used to
A steam bath looks like the ‘single hole’ water bath except that there is no
control the temperature of the liquid
heating element and no constant level device. Steam baths require piped steam
in it (usually water, but not always) to
as their source of heat, usually provided by a steam line, which is a permanent
within 0.5° C.
supply in the laboratory.
Beakers and conical flasks sit firmly on the rings, while round-bottom and
pear-shaped flasks should have about half the surface of the flask immersed in
Safety Note If you need to
the steam (Fig. 10.11).
increase or decrease the size of the
The main advantage of a water bath is the minimal risk of fire, while
‘hole’ by adding or removing rings, do
the major disadvantages are the maximum temperature available (∼100°C)
it before you start heating or use tongs
and the special precautions needed if anhydrous reaction conditions are
to prevent scalding by the steam.
required: remember that steam will condense down the inside of reflux
condensers.
Using a ‘multiple hole’ water bath –
make sure all the other ‘holes’ are cov-
ered otherwise the steam will escape
through them and heating your flask
will be very slow.
Safety Note Steam lines are very

dangerous and you should not attempt
to use them without direct supervision
from your laboratory instructor. water out
Safety Note Diethyl ether (ether) clamp

and hydrocarbons (petroleum spirit)
must never be heated on a hot plate. flask sitting clamp
water in
Diethyl ether (b.pt. 35 °C) should always on top of bath
flask -13 – -12
be heated under reflux using warm electric power immersed
in bath
water and petroleum hydrocarbons water out
should be heated under reflux on a
water bath or oil bath. water in water out
electric power water in
Using hot plates – never wrap the

power cable round the heating plate. It (a) (b)
may be hot and melt the flex, exposing
the wires. Fig. 10.11 (a) Heating a conical flask on a water bath; (b) heating a reflux
set-up on a water bath.

Hot plates and stirrer hot plates

Safety Note Using oil baths – oil
baths should be used in the fume cup-
These consist of a flat metal or ceramic plate, which is heated electrically, and
board because prolonged heating may
varies in size for use by an individual or by several people at the same time.
cause unpleasant and possibly toxic
The small versions normally have a built-in magnetic stirrer, which can be used
fumes to be released from the oil.
to stir the liquid with a magnetic ‘flea’.
Hot plates of both types should only be used for heating flat-bottom ves-
sels such as beakers or conical flasks and only when the liquid being heated is
non-flammable. The vapour of a flammable liquid may ‘run’ down the outside
Stirring in an oil bath – the stirrer hot-
of the vessel and ignite on the hot metal of the heating surface. Since the contact
plate will stir both the ‘flea’ in the oil
area between the heating surface and a round-bottom flask is very small, your
bath and the ‘flea’ in the reaction vessel.
attempts to heat these flasks effectively will require excessive temperatures of
the heating surface, which increases the dangers due to lack of control of the
heating process. Therefore round-bottom flasks should only be heated using
an oil bath (below) or a mantle (p. 91).
The flat exposed heating plate is extremely dangerous when hot: always
clamp
check that the plate is cool by passing your hand over the plate without touching
it or by placing a drop of water on the plate. If you have to pick up a hot plate,
hold it by the sides and do not touch the plate; it may burn. Typical uses of hot
plates are illustrated in Fig. 10.12a.
magnetic stirrer bar

Oil baths
These are mostly used to heat round-bottom flasks at temperatures above 100 °C.
heating plate The oil bath, containing the heating fluid, is usually a non-ferrous metal or
Pyrex® dish and heated on a stirrer hot plate, and the temperature of the bath is
variable speed control measured with a thermometer. The oil bath should never be more than half-full,
variable heating control to allow a margin of safety for thermal expansion of the oil, and stirred with
a magnetic ‘flea’ to ensure even heating. The equipment used in a typical oil
(a) bath is shown in Fig. 10.12b.
The nature of the oil used in the bath depends upon the temperature range
water out required and a selection of liquids is shown in Table 10.1.
You are most likely to encounter paraffin oil baths during your laboratory
work and the following safety points should be considered:
●● Paraffin oil discolours rapidly with prolonged heating. If the oil is dark,
thermometer clamp replace it with fresh oil. Dispose of the old oil in an appropriate manner
(check with laboratory staff).
●● Check that there is no water in the bottom of the oil bath: look for a separate
magnetic
water in stirring bars
layer or globules of liquid in the bottom of the bath. If you heat the bath
above 100 °C, the water will boil and may spatter hot oil over you and the
oil
Table 10.1 Oil bath liquids
Material Usable range (°C) Comments
Paraffin oil (mineral oil) 0–200 Flammable, cheap, produces

acrid smoke above 220°C
Ethylene glycol 0–150 Flammable, cheap, low flash
point
(b)
Polyethylene glycol 400 0–250 Water soluble
Fig. 10.12 (a) A stirrer hot plate. (b) Using Silicone oil 0–250 Expensive
a stirrer hot plate with an oil bath to heat a
Glycerol 0–250 Water soluble
round-bottom flask.

apparatus. Dispose of the contaminated oil into the appropriate waste con-
tainer, clean the bath with paper towels to absorb the water and refill with
fresh oil once completely dry.
●● If the oil bath is a Pyrex® dish, check the dish carefully for cracks and ‘star’
cracks because you do not want the dish of hot oil to break.
heat control ●● Support the stirrer hot plate on a ‘labjack’ so that you can quickly remove the
heat source if the reaction goes wrong (see Electric heating mantles, p. 91).
(a)
●● When the heating process is finished, allow the bath to cool and raise the
flask, let the oil drain from it into the bath and then wipe the flask with a
cloth or paper towel. Otherwise your hands, gloves, apparatus, compounds,
etc. may become contaminated with oil.
Electric heating mantles

heat control
These are used for heating round-bottom flasks only and come in various
sizes. Always use a heating mantle appropriate to the size of the flask you
stir control are going to heat, since you need to control the heating process. The flask
(b)
should fit snugly into the mantle with the top half of the flask above the case
of the mantle. If the mantle is too small, heating will be inefficient, whereas
Fig. 10.13 Mantles: (a) heating mantle; (b) if the mantle is too big, the flask will be overheated and decomposition of the
stirrer mantle. contents may occur.
Electric heating mantles comprise an electric heating element wrapped in
Safety Note Never connect an a glass fibre covering, protected by an earth screen and enclosed in an alumin-
electric heating mantle, requiring an
ium or, more commonly, a polypropylene case to allow handling at moderate
external controller, directly to the mains
temperatures. Heating control is provided either by a regulator built into the
electricity supply.
heating mantle (Fig. 10.13a) or by an external controller, which is connected
to the mantle by a plug.
Some mantles, ‘stirrer mantles’, have a built-in magnetic stirrer just like
a stirrer hot plate and can be used to mix the liquid using a magnetic flea or
bar (Fig. 10.13b). Stirrer mantles have two controls on the side: make sure that
you know the function of each, since one controls the extent of heating and the
other the stirrer speed.
water out When using mantles make the following safety checks:
clamp
●● Make sure that the heating element is not damaged or worn. If in doubt
consult your instructor and get a replacement.
water in ●● Plug in and test the mantle controls ensuring that both heater and stirrer are
working. If fumes are given off from the heating elements someone has spilt
mantle raised chemicals into the mantle: switch off, report the fault to your instructor and
and lowered
obtain a replacement.
●● Mantles are relatively slow to react to changes in the heater control setting
and it is easy to ‘overshoot’ the desired temperature. Therefore always make
small incremental changes in the heating control and if a temperature below
the boiling point of the liquid is required, make sure that a thermometer is
adjustable incorporated in the apparatus.
laboratory
jack ●● When using a mantle with complex apparatus such as for distillation (p. 145),
support the mantle on a ‘labjack’ so that it can be removed quickly if over-
heating occurs (Fig. 10.14).
to electrical supply
●● If the mantle is not equipped with a stirrer, remember to add anti-bumping
Fig. 10.14 Heating using a stirrer mantle. granules (p. 87) to the liquid.

Hot-air guns
These can be used instead of Bunsen burners or microburners provided that
the temperature required is not too high. The main uses of hot-air guns are for
drying glassware and as a heat source for distillation of liquids at relatively
moderate temperatures up to about 120°C.
Hot-air guns produce heated air, usually at two temperatures, and cold air.
As with a burner, the heat transferred to the flask being heated can be controlled
by ‘waving’ the hot air stream around the flask (see p. 88).
Remember that the hot-air gun has a hot-wire heating element; therefore
do not use it in the presence of flammable vapours. The nozzle of the hot-air
gun will become very hot and can cause burns or even ignite some solvents
for some time after it has been switched off. You should switch the gun to the
cold-air mode for a few minutes after you have finished heating and place it in
a ‘holster’ made from a support ring before finally turning off the gun.
Fig. 10.15 Rubber ‘fingers’.
Handling hot glassware
The safety precautions necessary for handling hot glassware depend upon:
Safety Note Never attempt to ●● The temperature of the glassware.
lift and manipulate a volume of liquid/
solution that you cannot carry easily; if
●● The type of glassware: test tube, beaker, conical flask, round-bottom flask,
in doubt divide equally between two
etc.
flasks. ●● The size of the glassware.
●● The manipulation being carried out.

Normally you do not need any hand protection to handle glassware at tem-
peratures up to 50°C. For general-purpose work, such as removing glassware
from the drying oven, assembling hot glassware and manoeuvring hot beakers
and conical flasks to and from a burner, steam bath or hot plate, heat-resistant
gloves are suitable and should be available in the laboratory. Where more
intricate processes are required, such as hot filtration, then ‘rubber fingers’
made from medium-wall rubber tubing (Fig. 10.15) give adequate protection
up to about 120 °C and are less cumbersome than insulated gloves. ‘Rubber
fingers’ are useful for small volumes, up to 150 mL, of liquids when the flask
or beaker can be easily held by one hand. If larger volumes are being manip-
Fig. 10.16 Holding a test tube. ulated then two hands are required and heat-resistant gloves are essential.
The following specific techniques should be noted:
●● Test tubes: should be held by a wooden test tube holder (Fig. 10.16), which
provides adequate insulation and grip. You should never use folded pieces
of paper or metal tongs.
●● Conical flasks: are often clamped to a support stand during heating and you
should never attempt to use the clamp as a device to hold the flask when
removed from the support stand. If you place the flask on the laboratory
bench with the clamp attached it will fall over because of the weight of the
clamp. Furthermore, you will have little control over the pouring process.
Use ‘rubber fingers’ or an insulated glove and never use folded pieces of
paper or metal tongs (Fig. 10.17).
●● Beakers: these have specific problems since they have no narrow neck
which can be gripped for lifting. Small beakers of volumes up to 400 mL
capacity can easily be gripped in one hand protected by an insulated glove
Fig. 10.17 Pouring a hot solution. or ‘rubber fingers’.

●● Round-bottom flasks: small flasks of capacity up to 250 mL should be held

at the neck, gripped in one hand protected by an insulated glove or ‘rubber
fingers’ and when moving and pouring from larger flasks they should be held
by the neck and supported underneath. Do not use a clamp round the neck
of the flask as a support.
Cooling
Cooling reactions – the heat from exo-
thermic reactions is absorbed by the During laboratory work you will be required to carry out experiments at tem-
cooling medium, otherwise the reaction peratures below room temperature. The most common situations where cooling
rate will increase rapidly with the rising is required are:
temperature and may result in violent
●● Cooling solutions during recrystallisation.
boiling or even explosion.
●● Completion of precipitation in quantitative (gravimetric) analysis and
preparations.
●● Cooling exothermic reactions.
●● Carrying out reactions at low temperatures.
There are three cooling media commonly used in the laboratory: crushed
ice, solid carbon dioxide (Dry Ice®, Drikold®, Cardice®) and liquid nitrogen.
You are unlikely to use liquid nitrogen in the undergraduate laboratory, if it is
required you must consult your demonstrator about the special safety protocols
required for its use.
Controlling temperature – temperature The most suitable containers for cooling baths are plastic bowls (ice baths),
control may be necessary to achieve the Pyrex® dishes (solid CO2 baths) and Dewar flasks (solid CO2 and liquid nitro-
desired reaction. For example, chemi- gen baths). If Pyrex® dishes are to be used below -20 °C, an insulated con-
cals may react to give different products tainer can be prepared by placing a smaller Pyrex® dish inside a larger one and
at different temperatures. filling the space between with an insulating material such as cotton wool, cork
chips or polyurethane foam chips. Remember that foam chips will dissolve if
they come into contact with many organic solvents.
If the temperature of the liquid or solution being cooled is critical, do not
assume that the temperature of the liquid or solution is the same as that of the
cooling bath: place a thermometer in the flask and remember that for tempera-
tures lower than -5 °C you should use an alcohol thermometer (red thread) or
a thermocouple-type thermometer (after checking that the probe will not react
with the contents of the flask).
Ice baths
A slurry of crushed ice and water can be used to give a cooling bath in the range
0 °C to 5 °C. Pure crushed ice does not give good contact with the glassware
and inefficient cooling results.
If temperatures below 0 °C are required, mixtures of crushed ice and
various inorganic salts can be used as shown in Table 10.2. Note that these
mixtures contain no liquid and therefore cooling is inefficient and the temper-
Table 10.2 Ice–salt mixtures atures indicated in the table are the lowest attainable under ideal conditions.
Salt Ratio (salt:ice) Temperature
Solid CO2 baths
CaCl2.6H2O 1:2.5 - 10 °C Solid CO2, when mixed with organic solvents, provides cooling media of tem-
NH4Cl 1:4 - 15 °C peratures ranging from -15 °C to -100 °C. Some common mixtures together
NaCl 1:3 - 20 °C with the minimum achievable temperatures are shown in Table 10.3. The CO2
and the organic solvent form a ‘slush’, which gives excellent contact with
CaCl2.6H2O 1:0.8 - 40 °C
flasks and, therefore, efficient cooling.

Table 10.3 Solid CO2 – solvent mixtures
Solvent Temperature Comments
Ethylene glycol - 15 °C Ice/NH4Cl cheaper

Acetonitrile - 40 °C Toxic, flammable
Chloroform - 60 °C Toxic
Ethanol - 72 °C Flammable
Acetone - 78 °C Flammable
Diethyl ether - 100 °C Highly flammable
Solid CO2 is supplied as large hard blocks or small quantities can be pre-
pared from cylinders of liquid CO2 as a ‘snow’. Skin contact with solid CO2
will cause frostbite and it must be handled with insulated gloves. The cooling
bath must be an insulated container, which will need to be topped up with solid
CO2 at regular intervals to maintain the temperature, or, if prolonged cooling
is required, a Dewar flask in which the coolant will maintain its temperature
for 12 hours or so.
To prepare a solid CO2 cooling bath:
●● Choose the appropriate solvent and remember to take into account the haz-
ards associated with its use.
●● Break the solid CO2 into small pieces. The CO2 ‘snow’ can be broken with
a spatula, but the hard blocks should be wrapped in cloth and then broken
into pieces with a wooden or polyethylene mallet.
●● Half fill the bath with the solvent and then, using an insulated glove, add
small pieces of the solid CO2 until the mixture stops ‘boiling’ and then add
a little more solid CO2 and stir with a glass rod to give a slurry.
Safety Note If the cooling bath
is more than half-full of solvent, it will ●● Use an alcohol or thermocouple thermometer to check the temperature of
‘boil’ out of the bath as you add the first the bath.
few lumps of solid CO2.
●● Top up the cooling bath with solid CO2 if the temperature begins to rise.
Key Point When using an internal thermometer to measure

the temperature of a liquid or solution which is being stirred
with a magnetic flea or stirring bar, ensure that the thermometer
bulb does not come into contact with it.
To prevent the condensation of water into your flask, you should have
Using cooling baths – remember that
an inert gas flowing through it (see p. 166) and prepare the cold bath around
a cooling bath will condense water from
the flask and its contents so that slow cooling occurs. Sudden immersion of
the atmosphere and therefore lose its
a relatively ‘hot’ flask into the cold bath will cause the bath to ‘boil’ and air
effectiveness over a period of time.
(containing water vapour) will be sucked into the apparatus despite the inert
atmosphere. Alternatively a ‘loosely packed’ CaCl2 guard tube (see p. 157) will
suffice if cooling is not too rapid.
Cooling probes
These are rigid or flexible metal probes, which are connected to a refrigeration
compressor. The probe is placed in the cooling bath and covered with a suitable
solvent, which is then cooled to the temperature desired. Cooling probes are

commonly found in constant temperature baths, when a temperature below

ambient temperature is required, and probes can be used instead of solid CO2 to
produce temperatures down to -100 °C. Cooling probes are expensive pieces
of equipment; therefore you will find them dedicated to a specific experiment
and they are not usually available for basic laboratory operations.
Drying
During your laboratory course it will be necessary to dry glassware, analyti-
cal standard compounds (see p. 192), chemicals you have synthesised, cruci-
bles used in gravimetric analysis (see p. 184) and solvents. Drying solvents is
described on p. 97.
Drying glassware
Safety Note Never place chem- For most general laboratory applications glassware can be dried in an electric
icals in the oven on a piece of filter oven between 80 °C and 90 °C or by rinsing the glassware with a small amount
paper. If your chemical melts it will run of water-miscible solvent, such as acetone or ethanol, and then evaporating
through the paper and may contami- the solvent using a compressed-air jet. Remember to remove all plastic com-
nate the samples of fellow students. If it ponents, taps and stoppers from the glassware before you put it in the oven.
is on a watch-glass it may be recovered If glassware is required for anhydrous reactions, it must be heated in the
if it has not decomposed. oven above 100 °C, assembled while hot and allowed to cool while a stream of
inert gas is passed through it (see p. 166).
Drying solids
Here the term ‘drying’ means removal of a solvent, not specifically water, from
a solid by evaporation. The rate of evaporation and thus the rate of drying can
be increased by one (or all) of the following:
●● Heating the chemical.
Safety Note Using corrosive acids
P2O5 and concentrated H2SO4 should ●● Using a drying agent in a closed container to absorb the solvent.
be avoided unless there is no suitable
●● Reducing the atmospheric pressure.
alternative. Consult your instructor for
their disposal. Only chemicals which are thermally stable should be dried by heating.
Most inorganic compounds, which are salts with relatively high melting points,
can be dried in an electric oven to remove water, whereas organic compounds,
many of which have relatively low melting points, need to be treated with more
care and the oven temperature should be set between 30 °C and 50 °C below
the melting point of the chemical. Chemicals must be placed in the oven on a
clock-glass or watch-glass and be spread as thinly as possible, to increase the
glass rate of solvent evaporation.
perforated plate If you cannot dry your compound in the oven, then use a desiccator. Des-
or gauze iccators are made from glass or plastic and some, vacuum desiccators, are
desiccant equipped with a tap to allow evacuation as shown in Fig. 10.18. The bottom
of the desiccator is filled with a drying agent (desiccant) and the chemical, on
a watch-glass or clock-glass, is placed on the mesh shelf above and the desic-
cator closed by sliding the lid onto the desiccator to provide an air-tight seal.
plastic The desiccant absorbs the solvent from the ‘atmosphere’ in the desiccator as it
evaporates from the solid. The nature of the desiccant depends upon the solvent
to be removed (Table 10.4).
The most common drying agents for removal of water are anhydrous CaCl2
vacuum tap
and self-indicating silica gel. The CaCl2 should have the appearance of ‘chalky’
lumps. Self-indicating silica gel is blue in the ‘active’ state and pink when its
Fig. 10.18 Desiccators. capacity for water absorption is ‘exhausted’. The water can be removed from

Table 10.4 Drying agents for desiccators
Solvent to be removed Drying agent Comments
H2O Silica gel

CaCl2
Solid KOH Corrosive
P2O5 Corrosive
Conc. H2SO4 Corrosive liquid
EtOH, MeOH CaCl2
Hydrocarbons Paraffin wax
the pink silica gel by heating in an oven above 100°C and restoration of the blue
Using vacuum desiccators – never
colour indicates reactivation.
switch off the vacuum supply before
The rate of drying can be increased by evacuating the desiccator and vac-
disconnecting the pipe from the des-
uum desiccators are specially designed for this purpose. The principal steps for
iccator since you will suck water or oil
the use of a vacuum desiccator are as follows and it is essential that you follow
into the supply pipe (see p. 86).
the order of the operations:
●● Check that the appropriate desiccant is present and active.
●● Check that the desiccator seals perfectly: ensure that the ground-glass edges
to the lid are greased lightly or, if the desiccator has a rubber gasket, carry
out a trial evacuation to ensure that the vacuum seals the desiccator by gently
pressing the lid onto the gasket – listen for air being sucked around the seal.
Using vacuum desiccators – never ●● Place the sample onto a watch- or clock-glass or a beaker covered with tissue
disconnect the vacuum supply with the paper (secure with an elastic band) and place it on the shelf.
tap on the desiccator open. Air will rush
●● Place the lid on the desiccator and open the tap and cover the desiccator with
into the desiccator and blow your dry
an appropriately sized safety cage (Fig. 10.19) to prevent injury from flying
compound around the inside.
glass in the case of an implosion.
●● Connect the tap to an operating source of vacuum: a water pump (p. 86) or
vacuum pump (p. 148) and open the tap slowly to evacuate the desiccator.
vacuum
stopcock
hole for
to vacuum
stopcock
perforated
shelf
desiccant
Fig. 10.19 Vacuum desiccator (with mesh safety cage).

●● When drying appears complete, close the tap on the dessicator and then
Environmental note Using water
disconnect the vacuum supply.
pumps for prolonged drying – this uses
too much water: evacuate the desicca- ●● Place a small piece of filter paper on the end of the tap and open the tap
tor, close the tap and disconnect and slowly. The filter paper will stick to the tap as air is drawn slowly through
turn off the water pump. Check periodi- it. When the air has completely filled the desiccator, the filter paper will fall
cally that the vacuum is preserved using off and it is safe to open the desiccator.
the filter paper technique. Re-evacuate
If you need to heat the compound under vacuum, then you will need to use
if necessary.
a vacuum oven (for large quantities of solids). The principles of operation of
these pieces of equipment are similar to those of a vacuum desiccator, except
that an electric heater is incorporated. Always allow the apparatus to cool to
Drying solvents – the drying agent
room temperature before releasing the vacuum.
used for solutions or pure liquids is not
usually suitable for drying solvents for Drying liquids
inert atmosphere reactions. This usually means removing water from a liquid chemical or a solution of a
chemical in a water-immiscible solvent. You will always need to dry solutions
after a liquid – liquid extraction (p. 139) and you may need to dry liquids after
Molecular sieves – are synthetic cal- evaporation (p. 161) or distillation (p. 145). In both cases the liquid is placed
cium and sodium aluminosilicates, in direct contact with the solid drying agent, i.e. the drying agent is added to
which have ‘holes’ of specific sizes to the liquid or solution. Ideally the drying agent should be totally insoluble in the
allow the absorption of molecules of liquid, should not react with it, absorb the water quickly and efficiently, and be
similar dimensions. Types 3A and 4A, easily filtered off. A list of the common drying agents is given in Table 10.5.
with 0.3 nm and 0.4 nm pores respec- You must remember that the drying agent will absorb some of the liquid or
tively, are used for drying. solvent being dried as well as the water. If you wish to dry a small volume of
Table 10.5 Drying agents for liquids and solutions

Drying solutions and liquids – note
that phrases such as ‘drying over mag- Drying agent Capacity Speed Efficiency Comments
nesium sulphate’ mean that the MgSO4 MgSO4 High Fast Good Best general use
is added to the solution.
Na2SO4 High Slow Poor Useful
CaCl2 High Slow Poor Reacts with O and N compounds
CaSO4 Low Fast Good Useful
K2CO3 High Fast Good Reacts with acidic compounds
Molecular sieve* Moderate Fast Good Must be activated at 300°C
Capacity: amount of water taken up.

Speed: rate of water absorption.
Efficiency: extent of drying after treatment.
*Do not throw away. Can be regenerated by heating 7 300 °C.
liquid, it is better to dissolve it in a low-boiling water-immiscible solvent and

dry the solution by the procedure described in Box 10.3.
Jointed glassware
This type of glassware, commonly known as Quickfit®, comprises a complete
range of components fitted with standard-taper ground-glass joints. The joints
are fully interchangeable with those of the same size and apparatus for a whole
range of experiments can be assembled from the simple components without
the need to use rubber bungs, corks, etc. Where there is a mismatch between the
sizes of the joints of the pieces of glassware, reduction and expansion adapters
can be used. A typical range of jointed glassware is illustrated in Fig. 10.20.

Box 10.3 How to dry a solution over magnesium sulphate
1. Place the solution to be dried in a clean, dry conical 4. Gravity filter the dried solvent layer through a fluted
flask. The flask should not be more than half-full. filter paper into a clean, dry flask. The fluted filter
paper should contain a small amount of fresh MgSO4,
2. Add small quantities of MgSo4 (between 0.1 g and as a safety precaution, in case a few drops of water are
1.0 g depending upon the volume of solvent) and floating in the surface of a denser solvent than water.
swirl the conical flask between each addition. At
first, the MgSO4 will ‘stick’ to the sides and bottom 5. Rinse the MgSo4 in the conical flask with a few milli-
of the flask where it contacts with the water, but on litres of pure dry solvent and filter it, to ensure that
further additions it will eventually form a free-flowing you recover all the solute, and then rinse the MgSO4
powder in the liquid and the solution will appear very on the filter paper.
clear and bright.
6. Remove the solvent by rotary evaporation (p. 162)
3. Allow the mixture to stand for 10 minutes. or distillation (p. 148).
The ground-glass joint on the glassware is classified according to the diam-

Safety Note Do not attempt to eter of the joint at its widest point (internal diameter) and the length of the
use non-standard ground-glass joints ground-glass portion of the joint. Thus a 14/23 joint has a maximum internal
in the standard jointware, it will not fit diameter of 14 mm and a length of 23 mm. Other common joint sizes you will
correctly. For example, ground-glass frequently encounter are 19/26, 24/29 and 35/39. The joint size is always etched
stoppers from volumetric flasks do not into glass on the side of or near to the joint. For obvious reasons, joints are
seal Quickfit® separatory funnels. categorised as ‘female’ and ‘male’.
Care of jointed glassware

Jointed glassware is much more expensive than ordinary glassware because of
the precision required in fabricating the joints. If the joints ‘seize’ and cannot
be separated the glassware cannot be used again and you may have the problem
of a stoppered flask containing a volatile organic solvent, which somebody has
to open! If this happens, consult your instructor for help and further advice.
There are two main causes of ‘seized’ joints:
1. Using solutions of potassium hydroxide or sodium hydroxide in water or

Greasing joints – you must grease all other solvents, which attack the glass.
the joints when carrying out a vacuum
2. Trapping chemicals, including solids and solutions of solids, in the ground-
distillation.
glass joints.
If you are using jointed glassware with strong alkalis (NaOH, KOH), you
must grease the joints. In most cases a simple hydrocarbon-based grease, such
Separating joints – always separate as petroleum jelly, will suffice, since it is easily removed from the joints by
ground-glass joints as soon as you have wiping with a cloth wet with a hydrocarbon solvent (petroleum spirit, b.pt.
finished with the apparatus and wipe 60–80°C). Avoid silicone-based grease, since this is difficult to remove, sol-
the joints clean. uble in some organic solvents and may contaminate your reaction products.
To grease a joint, put a small smear of grease on the upper part of the
‘male’ joint, push it into the ‘female’ joint with a twisting movement and the
joint should become ‘clear’ from the top to about half-way down. If more than
Environmental note – If you break a half the joint has become ‘clear’, you have used too much grease: separate the
piece of jointed glassware, do not throw joints, clean with a solvent-soaked cloth and repeat the process.
the item into the broken-glass bin. The To avoid trapping chemicals in the ground-glass joints, fill flasks, etc.
glass blower may be able to re-use it. using a long-stemmed filter funnel or paper cone, which extends past the joint
into the flask (see p. 80).

round-bottom flasks three-neck round-bottom flask
addition funnel separatory funnel
condenser air condenser

(or a fractionation column)
guard tube stopper reduction/expansion adapters
still-head splash-head Claisen adapter
distillation vacuum gas inlet thermometer

adapter distillation adapter adapter
adapter
Fig. 10.20 Glass equipment with standard-taper ground-glass joints.
Screwcap adapters
Screwcap or thermometer adapters allow you to place thermometers, glass
tubes or air bleeds into jointed glassware flasks. The screwcap adapter works
by using the screwcap to compress a rubber ring round the thermometer or glass

tube and thus hold it in place. The flexibility of the system allows the height of
screwcap
the thermometer/glass tube to be varied. The adapters come in different joint
sizes and varying hole sizes to accommodate different-diameter thermometers,
rubber ring
tubes, Pasteur pipettes, etc. The component parts of a screwcap adapter are
Teflon® seal
shown in Fig. 10.21.
To use the screwcap adapter with a thermometer:
●● Always disassemble the adapter to ensure that the rubber ring and the
Teflon® seal are present. If they are missing, get replacements before use.
The Teflon® seal is to protect the rubber ring from corrosive and solvent
vapours.
joint
●● Ease the rubber ring onto the thermometer (see p. 68) and slide the Teflon®
seal on below the ring.
Fig.10.21 The screwcap adapter.
●● Slide the screwcap over the top of the thermometer and then screw the whole
assembly onto the base of the adapter and tighten the screw slightly, just
enough to hold the thermometer.
Using screwcap adapters – if the rub- ●● Trial fit the adapter and thermometer into the apparatus and adjust the height
ber ring is not present in the screwcap of the thermometer by loosening the screwcap and carefully sliding the ther-
adapter, the thermometer will slide out mometer up or down as required. When satisfied with the fit, re-tighten the
of the adapter and may smash both the screwcap.
flask and the thermometer.
●● Check for final fit and tighten the screwcap firmly, but do not over-tighten.
Joint clips
Plastic joint clips or Keck® clips (Fig. 10.22a) are used for holding ground-glass
joints firmly together and may be used to replace clamps and support stands
at certain points when building apparatus (see p. 147) and are essential when
using rotary evaporators (p. 162). The main weakness of these otherwise use-
ful devices is that they soften at about 130 °C and this may allow the joints to
separate. Therefore they should never be used at the ‘hot end’ of a distillation,
for example. The clip should be used to hold a distillation adapter on the end
of a water condenser, or the flasks onto a fraction collector, but never on the
distilling flask or to hold the condenser onto the still head (Fig. 10.22b).
no joint clips here
joint clips here
water
out
water
joint clip or Keck® clip in
(a) (b)
Fig. 10.22 Joint clips and their use.

When using joint clips always:

●● Check that you are using the appropriate size of clip for the joints being held.
The clips are often colour coded.
●● Check that the clip is not cracked or split.
●● Check that the wide ‘lower jaw’ of the clip fits under the rim of the ‘female’
joint and the ‘upper jaw’ fits round the male joint.
●● Support the joints with your hand as you push the clip into place. If in doubt
use a protective insulated glove.

Brown, T., LeMay, H., Bursten, B., Murphy, C., Woodward, P., Mendham, J., Denney, R.C., Barnes, J.D. and Thomas,
Nelson, J. and Kemp, K. (2014) Laboratory Experiments M.J.K. (2000) Vogel ’s Textbook of Quantitative
for Chemistry, 13th edn. Prentice Hall, Harlow, Essex. Chemical Analysis, 6th edn. Prentice Hall, Harlow,
Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch- Essex.
ell, A.R. (1991) Vogel’s Textbook of Practical Organic McMurry, J.E., Fay, R.C., Robinson, J.K., Dillon, S. and
Chemistry, 5th edn. Longman, Harlow, Essex. Rogers, S. (2015) Laboratory Manual for Chemistry,
Harwood, L.M., Moody, C.J. and Percy, J.M. (1999) 7th edn. Pearson, Harlow.
Experimental Organic Chemistry, 2nd edn. Blackwell Pass, G. and Sutcliffe, H. (1979) Practical Inorganic
Science Ltd, Oxford. Chemistry. Kluwer Academic Publishers, Netherlands.
Isac-Garcia, J., Dobado, J.A., Calvo-Flores, F.G. and Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical
Martinez-Garcia, H. (2015) Experimental Organic Organic Chemistry. Chapman & Hall, London.
Chemistry. Laboratory Manual. Elsevier, Amsterdam. Suib, S.L. and Tanaka, J. (1999) Experimental Methods in
Leonard, J., Lygo, B. and Procter, G. (2013) Advanced Prac- Inorganic Chemistry. Prentice Hall, Harlow, Essex.
tical Organic Chemistry, 3rd edn. CRC Press, Boca Raton. Zubrick, J.W. (2015) The Organic Chem Lab Survival
Lehman, J.W. (2009) Student Lab Companion. Laboratory Manual. A student’s guide to techniques, 10th edn. John
techniques for organic chemistry, 2nd edn. Pearson, Wiley & Sons Ltd, Chichester.
Harlow.
Study exercises
10.1 Write down the apparatus you would need to carry (c) Removal of anti-bumping granules from a
out the following operations, giving your reasons: solution of N-phenylethanamide in hot water.
(d) Heating a suspension of N-phenylethanamide
(a) Separation of a mixture of soil and water to
in cold water so as to dissolve it for
allow you to measure the nitrate content of
recrystallisation.
the soil. Remember, nitrates are water soluble.
(e) Heating a solution in toluene (b.pt. 111 °C,
(b) Isolation of solid, water insoluble N-
flammable).
phenylethanamide from the reaction mixture
(f) Cooling a reaction mixture to -10 °C.
which comprises N-phenylethanamide and
very dilute ethanoic acid.

11 Principles of solution chemistry
A solution is a homogeneous liquid, formed by the addition of solutes to a sol-

Preparing solutions – practical advice
vent. The behaviour of solutions is determined by the type of solutes involved
is given on p. 71.
and by their proportions, relative to the solvent. Many laboratory exercises
involve calculation of concentrations, e.g. when preparing an experimental
solution at a particular concentration, or when expressing data in terms of sol-
ute concentration. Make sure that you understand the basic principles set out
Definitions in this chapter before you tackle such exercises.
Solutes can affect the properties of solutions in several ways, as follows.
Electrolyte – a substance that dissoci-
ates, either fully or partially, in water to
give two or more ions.
Electrolytic dissociation
Relative atomic mass (Ar) – the mass of This occurs where a substance dissociates to give charged particles (ions).
an atom relative to 12 C = 12. For a strong electrolyte, e.g. Na + Cl - , dissociation is essentially complete. In
Relative molecular mass (Mr) – the mass contrast, a weak electrolyte, e.g. ethanoic acid, will be only partly dissociated,
of a compound’s formula unit relative to depending upon the pH and temperature of the solution (p. 114).
12
C = 12.
Mole (of a substance) – the equiva- Ideal/non-ideal behaviour
lent in mass to relative molecular mass This occurs because solutions of real substances do not necessarily conform
in grams. Note this is a pragmatic rule to the theoretical relationships predicted for dilute solutions of so-called ideal
rather than a definition. solutes. It is often necessary to take account of the non-ideal behaviour of
real solutions, especially at high solute concentrations (see Haynes (2015) for
appropriate data).
Do not confuse the solubility of a
chemical with its strength as an elec- Concentration
trolyte. Ethanoic acid is completely
soluble with water in all proportions, yet In SI units (p. 40) the concentration of a solute is expressed in mol m -3, which
it is a weak electrolyte because it is only is essential for calculating specific parameters for substances (e.g. p. 43), but
partially dissociated. Barium hydroxide which is inconvenient when dealing with solutions in the laboratory. A cubic
is very insoluble in water, but the small metre (m3) of water weighs approximately 1 ton! A common unit of volume
quantity which does dissolve (see Ks used in chemistry is the litre (L): this is a non-SI unit and is converted to the SI
below) is dissociated completely into unit of volume (m3) using 1.0 L = 10-3 m3. The concentration of a solute is
Ba2 + and OH - ions; thus it is a strong usually symbolised by square brackets, e.g. [NaCl]. Details of how to prepare
electrolyte. solutions are given on pp. 72, 74.
A number of alternative ways of expressing the relative amounts of solute
and solvent are in general use, and you may come across these terms in your
practical work or in the literature.
Expressing solute concentrations –
you should use SI units wherever pos- Molarity
sible. However, you are likely to meet
This is the term used to denote molar concentration, [C], expressed as moles
non-SI concentrations and you must be
of solute per litre volume of solution (m o l L-1 ). This non-SI term continues to
able to deal with these units too.
find widespread usage, in part because of the familiarity of working scientists
with the term, but also because laboratory glassware is calibrated in millilitres
and litres, making the preparation of molar and millimolar solutions relatively
straightforward. However, the symbols in common use for molar (M) and
Example A 1.0 molar solution of
millimolar (mM) solutions are at odds with the SI system and many people
NaCl would contain 58.44 g NaCl (the
molecular mass) per litre of solution.
now prefer to use m o l L-1 and m m o l L-1 respectively, to avoid confusion.
Box 11.1 gives details of some useful approaches to calculations involving
molarities.

Principles of solution chemistry
Box 11.1 Useful procedures for calculations involving molar concentrations
1. Preparing a solution of defined molarity. For a solute where [C 1 ] and [C2 ] are the initial and final concentra-
of known relative molecular mass (Mr), the following tions, while V1 and V2 are their respective volumes:
relationship can be applied: each pair must be expressed in the same units. Thus,
if you wanted to dilute 200 mL of 0.5 mol L-1 NaCl to
mass of solute/ Mr give a final molarity of 0.1 mol L-1 , then, by substi-
[C] = [11.1] tution into eqn [11.2]:
volume of solution
0.5 * 200 = 0.1 * V2
So, if you wanted to make up 200 mL (0.2 L) of an
aqueous solution of NaCl (Mr = 58.44 g) at a con-
centration of 500 mmol L-1 (0.5 mol L-1 ), you could Thus V2 = 1000 mL (in other words, you would have
calculate the amount of NaCl required by inserting to add water to 200 mL of 0.5 mol L-1 NaCl to give
these values into eqn [11.1]: a final volume of 1000 mL to obtain a 0.1 mol L-1
solution).
mass of solute/ 58.44 3. Interconversion. A simple way of interconverting
0.5 =
0.2 amounts and volumes of any particular solution is to
which can be re-arranged to divide the amount and volume by a factor of 10 3 : thus
a molar solution of a substance contains 1 mol L-1 ,
mass of solute = 0.5 * 0.2 * 58.44 = 5.844 g which is equivalent to 1 mmol mL-1 , or 1 mmol mL-1 ,
or 1 nmol nL-1 , etc. You may find this technique use-
The same relationship can be used to calculate the ful when calculating the amount of substance present
concentration of a solution containing a known in a small volume of solution of known concentra-
amount of a solute, e.g. if 21.1g of NaCl were made tion, e.g. to calculate the amount of NaCl present in
up to a volume of 100 mL (0.1 L), this would give: 50 mL of a solution with a concentration (molarity) of
0.5 mol L-1 NaCl:
21.1/ 58.44
[NaCl] = = 3.61 mol L-1
0.1 (a) this is equivalent to a concentration of
0.5 mmol mL-1 ;
2. Dilutions and concentrations. The following rela- (b) t h e r e f o r e 50 mL will contain
tionship is very useful if you are diluting (or concen- 50 * 0.5 mmol = 25 mmol.
trating) a solution:
[C 1 ]V1 = [C 2 ]V2 [11.2] The ‘unitary method’ (p. 473) is an alternative

approach to these calculations.
Molality
Example A 0.5 molal solution of NaCl This is used to express the concentration of solute relative to the mass of sol-
would contain 58.44 * 0.5 = 29.22 g vent, i.e. m o l kg -1 . Molality is a temperature-independent means of express-
NaCl per kg of water. ing solute concentration.
Per cent composition (% w/w)
This is the solute mass (in g) per 100 g solution. The advantage of this expres-
sion is the ease with which a solution can be prepared, since it simply requires
Example A 5% w/w NaOH solution each component to be pre-weighed (for water, a volumetric measurement may
contains 5 g NaOH and 95 g water be used, e.g. using a measuring cylinder) and then mixed together. Similar terms
( = 95 mL water, assuming a density are parts per thousand (‰), i.e. mg g -1, and parts per million (ppm), i.e. mg g -1.
of 1 g mL-1) to give 100 g of solution.
Per cent concentration (% w/v and % v/v)
For solutes added in solid form, this is the number of grams of solute per
100 mL solution. This is more commonly used than per cent composition, since
solutions can be accurately prepared by weighing out the required amount of

solute and then making this up to a known volume using a volumetric flask.
Example A 5% w/v KOH solution con- The equivalent expression for liquid solutes is % v/v.
tains 5 g KOH in 100 mL of solution. A
The principal use of mass/mass or mass/volume terms (including g L-1 ) is
5% v/v glycerol solution would contain
5 mL glycerol in 100 mL of solution.
for solutes whose molecular mass is unknown (e.g. polymers), or for mixtures
Note that when water is the sol- of certain classes of substance (e.g. total salt in sea water). You should never
vent this is often not specified in the use the per cent term without specifying how the solution was prepared, i.e. by
expression, e.g. a 20% v/v ethanol using the qualifier w/w, w/v or v/v. For mass concentrations, it is often simpler
solution contains 20 mL ethanol made to use mass per unit volume, e.g. mg L-1, mg mL-1.
up to 100 mL of solution using water.
Parts per million concentration (ppm)
This is a non-SI weight per volume (w/v) concentration term commonly used
in quantitative analysis such as flame photometry, atomic absorption spec-
troscopy and gas chromatography, where low concentrations of solutes are to
Note that ppm may be a weight/ be analysed. The term ppm is equivalent to the expression of concentration as
weight (w/w) expression. The origin of mg m L-1 (1 0 -6 g m L-1 ) and a 1.0 ppm solution of a substance will have a
the term ppm derives from a solution concentration of 1 .0 mg m L-1 (1 .0 * 1 0 -6 g m L-1 ). A typical procedure
whose concentration is 1 ppm if it con- for calculations in terms of ppm is shown in Box 11.2.
tains 1 g of solute for each million (10 6 ) g Parts per billion (ppb) is an extension of this concentration term as
of solvent. n g m L-1 (1 0 -9 g m L-1 ) and is commonly used to express concentrations of
very dilute solutions. For example, the allowable concentration of arsenic in
water may be 0.05 ppm, but it is more conveniently expressed as 50 ppb.
Activity (a)
This is a term used to describe the effective concentration of a solute. In dilute
solutions, solutes can be considered to behave according to ideal (thermody-
namic) principles, i.e. they will have an effective concentration equivalent to
Box 11.2 How to convert ppm into mass of chemical required
Example: Suppose you are asked to prepare an aqueous (a) S o d i u m chloride (NaCl), Mr = 58.5:
solution of sodium ions (250.00 mL) of approximate, but therefore you need to weigh out
accurately known, concentration of 10 ppm from either 58.5 * 0.1087 * 10 -3 = 6.359 * 10 -3 g of
sodium chloride or anhydrous sodium carbonate. sodium chloride to be made up to 250.00 mL for
a 10 ppm solution of sodium ions.
1. Convert the ppm concentration into g L−1 . Thus
1.0 ppm = 1.0 * 10 -6 g mL-1 and hence a solution of (b) Sodium carbonate (Na2 CO3 ), Mr = 106: you
10 ppm = 10 * 10-6 g mL-1 = 103 * 10 * 10-6 g L-1 must note that each ‘molecule’ of sodium
= 10 * 10-3 g L-1. carbonate contains two sodium ions; thus
the number of moles of sodium carbonate
2. Convert the concentration from gL−1 to mol L−1 . The required for a 10 ppm solution of sodium ions
Ar for sodium ion is 23 g and for a litre of 10 ppm is 0.1087 * 10 -3 , 2 = 0.054 35 * 10 -3 mol.
solution you need 10 * 10 -3 , 23 = 0.435 * 10 -3 Yo u must therefore weigh out
mol of sodium ions. 106 * 0.054 35 * 10 -3 = 5.7611 * 10 -3 g of
sodium carbonate to be made up to 250.00 mL
3. Convert the number of moles per litre into moles in the for a 10 ppm solution of sodium ions.
volume required (250.00 mL). Since 1.0 litres of 10 ppm
solution of sodium ions requires 0.435 * 10 -3 mol of
sodium ions, then 250.00 mL (0.25 L) of solution will need 5. Decide how you are to prepare these solutions using
0.25 * 0.435 * 10 -3 mol = 0.1087 * 10 -3 mol of the procedures outlined in Boxes 11.1, 11.2 and 11.3,
sodium ions. since the calculation shows that you will need to
weigh out small masses of chemicals, which are at
4. Calculate the mass of either sodium chloride or the limit of accuracy of an analytical balance.
sodium carbonate required to make up the solution:

Table 11.1 Activity coefficient of NaCl solu- the actual concentration. However, in concentrated solutions (Ú 0.5 mol L-1),
tions as a function of molality. Data from the behaviour of solutes is often non-ideal, and their effective concentration
Robinson and Stokes (2012) (activity) will be less than the actual concentration [C]. The ratio between
the effective concentration and the actual concentration is called the activity
Molality Activity coefficient at 25 °C
coefficient (g) where
0.1 0.778
a
0.5 0.681 g = [11.3]
[C]
1.0 0.657
2.0 0.668 Equation [11.3] can be used for SI units (m o l m -3 ), molarity (m o l L-1 )
or molality (m o l kg -1 ). In all cases, g is a dimensionless term, since a and
4.0 0.783
[C] are expressed in the same units. The activity coefficient of a solute is effec-
6.0 0.986 tively unity in dilute solution, decreasing as the solute concentration increases
(Table 11.1). At high concentrations of certain ionic solutes, g may increase to
become greater than unity.
Example A solution of NaCl with

Key Point Activity is often the correct expression for theoret-
a molality of 0.5 mol kg -1 has ical relationships involving solute concentration (e.g. where a
an activity coefficient of 0.681 property of the solution is dependent on concentration). How-
at 25 °C and a molal activity of ever, for most practical purposes, it is possible to use the actual
0.5 * 0.681 = 0.340 mol kg -1 . concentration of a solute rather than the activity, since the differ-
ence between the two terms can be ignored for dilute solutions.
Equivalent mass (equivalent weight)

Equivalence and normality are outdated terms, although you may come across
them in older texts. The magnitude of an equivalent mass (equivalent weight)
can be simply identified from the balanced equation for the reaction being
considered. Remember that the equivalent mass can change, depending on the
reaction, as the following reactions illustrate.
For:
HCl + Na OH S Na Cl + H2 O
1 mol of HCl reacts with 1 mol of NaOH, the equivalent mass of HCl is
Mr = 3 6 .5 and the equivalent mass of NaOH is also its Mr = 4 0 .
For:
H2 SO 4 + 2 Na OH S Na 2 SO 4 + 2 H2 O
since 1 mol of H2 SO 4 reacts with 2 mol of NaOH, the equivalent mass of

H2 SO 4 is Mr , 2 = 9 8 , 2 = 4 9 , while the equivalent mass of NaOH is
still Mr = 4 0 .
For:
5 Fe SO 4 + KMn O 4 S Fe 2 (SO 4 )3 + 2 Mn SO 4
since 1 mol of KMn O 4 reacts with 5 mol of Fe SO 4 , then the equivalent mass
of KMn O 4 is Mr , 5 = 1 5 8 , 5 = 3 1 .6 , and that of Fe SO 4 is still
Mr = 1 5 2 .
But, for:
H2 SO 4 + Na 2 CO 3 S Na 2 SO 4 + CO 2 + H2 O
since the reaction is 1:1, the equivalent masses of H2 SO 4 and Na 2 CO 3 could

be their Mr values, 98 and 106 respectively.
As a result of this possible confusion, the concept of equivalent mass
(weight) is rarely used.

Normality
A 1 normal solution (1 N) is one that contains one equivalent mass of a sub-
stance per litre of solution. The general formula is:
m a s s o f s u b s ta n ce p e r litre
n o rm a lity = [11.4]
Insolubility – no solute can be shown e q u iva le n t m a s s
to be completely insoluble in a given The use of normality is now obsolete.
solvent, but for practical purposes, a
compound which has less than 0.01% Solubility
(w/w) solubility in a solvent can be con-
sidered to be insoluble in that solvent.
The extent to which a solute will dissolve in a solvent is called its solubility.
The solubility of a chemical is conventionally expressed as the maximum num-
ber of grams of a chemical that will dissolve in 100 g of solvent but conversion
to m o l L-1 or g L-1 is simple and may be appropriate for some applications
Variation of solubility – the solubility (see below). Since solubility is temperature dependent, it is always quoted at
of a chemical may vary in different sol- a specific temperature. With a very few exceptions, increasing the temperature
vents. For example, NaCl is soluble in of a solvent increases the solubility of the solute.
water but insoluble in DCM, whereas
for naphthalene the opposite is true. Saturated solutions
For practical purposes, a saturated solution is one in which no more solute will
dissolve. For example, the solubility of sodium chloride in water is 35.6 g per
100 g at 25 °C and 39.1 g per 100 g at 100 °C and both solutions are saturated
Saturated solutions – theoretically, a solutions at their respective temperatures. If the 100 °C solution is cooled to
saturated solution is one in which the 25 °C, then 3.5 g of NaCl crystals will precipitate from the solution, because
solution is in dynamic equilibrium with the solution at 25 °C requires only 35.6 g of NaCl for saturation. This process
the undissolved solute. is the basis of purification of compounds by recrystallisation (see p. 125).
Solubility product
In dilute aqueous solutions, it has been demonstrated experimentally for poorly
Solubility – remember to use
soluble ionic salts (solubilities less than 0 .0 1 m o l L-1 ) that the mathematical
mass = volume * density when con-
product of the total molar concentrations of the component ions is a constant
verting solubilities from grams of sol-
at constant temperature. This product, Ks is called the solubility product. Thus
ute per 100 g of solvent to g L-1 , when
for a saturated solution of a simple ionic compound AB in water, we have the
using solvents other than water.
dynamic equilibrium:
+ -
ABsolid ÷ A(aq) + B(aq)
where AB represents the solid which has not dissolved, in equilibrium with its
ions in the aqueous saturated solution. Then:
Ks = [A+ ] * [B- ]
For example, silver chloride is a solid of solubility 0.00015 g per 100 mL of

water in equilibrium with silver cations and chloride ions. Then:
Ks = [Ag + ] * [Cl- ]
The solubility of AgCl is 0 .0 0 1 5 g m L-1 (1 0 * solubility per 100 g, assum-

ing that the density of water is 1 .0 g m L-1 ) and therefore the solubility of AgCl
is 0 .0 0 1 5 , 1 4 3 .5 = 1 .0 5 * 1 0 -5 m o l L-1 . Thus the saturated solution
contains 1 .0 5 * 1 0 -5 m o l L-1 of Ag + ions and 1 .0 5 * 1 0 -5 m o l L-1 of
Cl - ions and the solubility product Ks is
Ks = (1 .0 5 * 1 0 -5 ) * (1 .0 5 * 1 0 -5 ) = 1 .1 * 1 0 -1 0 m o l2 L-2

If the solid does not have a simple 1:1 ratio of its ionic components, e.g.
Pb Cl2 , then the solubility product is given by:
Ks = [Pb2+ ] * [Cl- ]2
In general terms, the solubility product for a compound My Nx , is given by
Ks = [M + ]y * [N- ]z
The practical effects of solubility products are demonstrated in the detec-
tion of anions and cations by precipitation (p. 179) and in quantitative gravi-
metric analysis (p. 184). For example, if dilute aqueous solutions of silver
nitrate (solubility 55.6 g per 100 g of water) and sodium chloride (solubility
35.6 g per 100 g of water) are mixed, an immediate white precipitate of AgCl
is produced because the solubility product of AgCl has been exceeded by the
numbers of Ag + and Cl - ions in the solution, even though the ions come from
different ‘molecules’. A saturated solution of AgCl is formed and the excess
AgCl precipitates out. The solubility product of the other combinations of ions
is not exceeded and thus sodium and nitrate ions remain in solution. Even if the
concentration of Ag + is extremely low, the solubility product for AgCl can be
exceeded by the addition of an excess of Cl - ions, since it is the multiplication
of these two concentrations which defines the solubility product. Thus solu-
ble chlorides can be used to detect the presence of Ag + ions and, conversely,
soluble silver salts can be used to detect Cl - ions, both quantitatively and
qualitatively.
Reactions of ions in solution

There are essentially only four basic reactions of ions in solution:
1. Acid–base reactions.
2. Precipitation reactions.
3. Complexation reactions.
4. Reduction–oxidation (redox) reactions.
Acid–base, precipitation and complexation reactions are all examples of
exchange (metathesis) reactions in which ions in solution ‘exchange partners’,
for example:
A+ Y- + B + Z - S A+ Z - + B + Y-
Metathesis reactions are really equilibria between the ionic species, which
are displaced to the right (to the reaction product) by a feature which defines
the classification of the reaction type.
Acid–base reactions
The most common acid–base reactions are exemplified by the neutralisation
reaction between hydrogen ions and hydroxide ions; for example, the reaction
between dilute hydrochloric acid and dilute sodium hydroxide:
H + Cl(a
- + - + -
q ) + Na OH (a q ) S H@OH(liq ) + Na Cl(aq )
Since water is essentially a covalent compound (see p. 113) its formation

effectively removes H + and OH - from the equilibrium and drives the reaction
to completion. Other examples of this general type of reaction include the

removal of a molecule as a gas, such as reactions of acids with carbonates and

bicarbonates, where unstable H2 CO 3 decomposes to H2 O and CO 2 .
Precipitation reactions
In these reactions between ions, one substance is removed from the ionic equi-
librium by precipitation (see solubility product, p. 106) and drives the equilib-
rium to the right (see p. 106).
Complexation reactions
A complex ion is formed by the reaction of a metal cation, in particular transition
metals, with an electron donor molecule (ligand), which can be neutral or have a
negative charge. The cation can accept an electron pair and the ligand donates an
electron pair to form a covalent donor (co-ordinate) bond between the ligand and
the metal ion. The ligands are said to co-ordinate with the metal ion to give a com-
plex. Many ligands are more powerful electron donors than water and thus the
addition of a ligand to an aqueous solution of a metal cation displaces the equi-
librium towards the more stable complex ion (see p. 203 for stability constants
and complexes). The effects of complex formation are illustrated in Box 11.3.
The overall effect of complex formation is to ‘remove’ a hydrated metal
ion from the mixture of ions in solution by displacing the equilibrium in favour
of the complex, cf. the similar process in the formation of water in acid–base
titrations and precipitation reactions.
Reduction–oxidation (redox) reactions
The concepts of oxidation and reduction are defined in terms of complete elec-
tron transfer from one atom, ion or molecule of a chemical to another:
●● Chemical oxidised – chemical loses electron(s).
●● Chemical reduced – chemical gains electron(s).
Box 11.3 An example of complex formation
If an aqueous solution of ammonia is added to an aque- 3. As the ammonia solution is added, the solubility
ous solution of copper (II) sulphate the following changes product of Cu(OH)2 is exceeded, even by the low
are observed. concentration of OH - ions and the white solid,
Cu(OH)2 , precipitates from the saturated solution
A. On addition of the ammonia solution to the pale-blue (see p. 106).
copper solution a white precipitate forms.
4. As addition of the ammonia solution is continued,
B. As addition is continued, the white precipitate dis- the free NH3 molecules displace the water mole-
solves and a royal-blue solution is formed, which does
cules from the pale-blue Cu(H2 O)24 + complex ion to
not change on further addition of ammonia solution.
form the royal-blue Cu(NH3 )24 + complex, which is
These changes can be explained as follows: more stable than the water complex (larger stability
constant).
1. Ammonia solution is an equilibrium mixture, which
lies well to the left. Consequently a dilute solution of 5. Since the insoluble Cu(OH)2 is in equilibrium with the
ammonia contains a little OH - and lots of free NH3 : Cu(H2 O)24 +, which is being removed as the Cu(NH3 )24 +
complex, the Cu(OH)2 reverts to Cu(H2 O)24 + which
NH3 + H2 O ÷ NH4+ + OH - then forms the Cu(NH3 )24 + complex. Thus the white
precipitate dissolves leaving the royal-blue solution
2. Copper (II) sulphate comprises the Cu(H2 O)24 + ion and of the Cu(NH3 )24 + complex.
SO24 - ions in solution.

This approach is generally applicable to most reactions and avoids compli-

cations of the older definitions involving hydrogen and oxygen. You should
realise that if a chemical is oxidised during a reaction, then another must
be reduced: oxidation and reduction always occur together. Furthermore:
●● Oxidising agent – gains electron(s) and is therefore reduced.
●● Reducing agent – loses electron(s) and is therefore oxidised.
The following reaction between magnesium metal and dilute acid illus-
trates these concepts:
+ 2+
Mg (s ) + 2 H (a q ) S Mg (a q ) + H2 ( g )
Magnesium metal has lost two electrons in forming Mg 2 + ions and has there-
fore been oxidised. The two protons have each gained an electron to form
hydrogen atoms (and then one hydrogen molecule) and have been reduced.
Since magnesium metal has been oxidised, it is a reducing agent and because
H + has gained an electron, it is an oxidising agent.
The stoichiometry of a redox reaction is defined by the number of electrons
transferred between the oxidising agent and the reducing agent since the num-
ber of electrons lost by the reducing agent must equal the number of electrons
gained by the reducing agent, e.g.
2 Mg (s ) + O 2 ( g ) S 2 Mg O (s )
So that you can work out titrations involving redox reactions, you will
find it necessary to balance redox equations, and while it is easy for simple
reactions such as those above, more complex redox reactions, such as the one
below, require more thought and work.
2KMnO4 + 5H2O2 + 3H2SO4 S 2MnSO4 + K2SO4 + 5O2 + 8H2O

Such problems can be broken down into several simple steps, each with its
own set of rules:
●● Identify the atoms, ions or molecules which have been oxidised and reduced.
●● Identify the ionic half-reactions for the species being oxidised and reduced
and combine them.
●● Balance the ionic half-reactions and combine them to give a balanced equa-
tion for the reaction.
The species which are oxidised and reduced can be identified using the
concept of oxidation numbers. The rules for determining oxidation num-
bers and examples are given in Box 11.4 and the application of ionic half-
reactions to balance redox equations is shown in Box 11.5. Note that the
result of the use of partial ionic equations gives a balanced ionic equation
for the redox reaction.
Key Point In simple acid–base, precipitation and complexa-

tion reactions, no change of oxidation number occurs at any of
the atoms involved.

Box 11.4 The use of oxidation numbers to identify redox systems
The oxidation number is a hypothetical charge assigned (rule 2), four oxygen atoms, 4 * -2 = - 8 (rule 3b);
to atoms in molecules and ions using a set of specific therefore Cl must be +7 (rule 3c).
rules. Since redox reactions involve transfers of electrons,
identification of the atoms which change oxidation num- 5. The sum of the oxidation numbers of all the atoms
ber will show the atoms, ions or molecules which are in a polyatomic ion is equal to the charge on the ion,
specifically involved in the redox process. e.g. in CO23 - ion, each oxygen is -2 (rule 3b); thus
Rules for oxidation numbers 3 * -2 = - 6. Since the charge on the CO23 - ion is
2 - , then carbon must be +4.
1. For an atom in its elemental form, the oxidation
number is always 0. Thus Cl in Cl2 has an oxidation 6. If the oxidation number of an atom becomes more
number 0, as does Na metal, and carbon in charcoal, positive during the reaction, it has lost electrons and
graphite or diamond. has been oxidised.
2. For any monatomic ion, the oxidation number is the
same as the charge on the ion. Thus Na+ has an oxida- If the oxidation number of an atom becomes more
tion number of + 1, Cl - is - 1, Al3+ is +3, S2- is -2, etc. negative during the reaction, it has gained electrons
and has been reduced.
3. Non-metals usually have negative values, but there
are some exceptions: Example: If you consider the unbalanced equation for the
reaction shown on p. 109:
(a) The oxidation number of fluorine is always -1 in
KMnO4 + H2O2 + H2SO4 S MnSO4 + K2SO4
all compounds.
+ O2 + H2O
(b) The oxidation number of oxygen is always -2,
except when bonded to fluorine (OF2 ), in perox- you can now calculate that the only atoms which
ides (O22 -), where each oxygen atom is -1 and have changed oxidation number are manganese,
superoxides (O2-), where each oxygen atom is - 12 . which has changed from +7 in MnO4- to + 2 as Mn2 +
(c) The oxidation number of the other halogens is in MnSO4 , and oxygen, which has changed from - 1
always - 1, except when bonded to atoms of in H2 O2 (rule 3b) to 0 in O2 . Thus Mn has gained elec-
greater electronegativity, e.g. Cl in ClF3 is +3, Br trons and been reduced and oxygen has lost electrons
in BrCl is + 1, etc. and been oxidised. Furthermore, the Mn atom has
gained five electrons and the O22 - ion has lost two
(d) The oxidation number of hydrogen is always +1, electrons, so five molecules of H2 O2 should react with
except when bonded to electropositive metals, two molecules of KMnO4 .
when it is - 1, e.g. in HCl it is +1 in NH3 it is +1 The two partial ionic reactions can now be identified:
and in NaH, MgH2 and AlH3 it is -1.
For reduction MnO4- S Mn2 +
4. The sum of the oxidation numbers of all atoms in For oxidation H2 O2 S O2
a neutral compound is zero, e.g. in KClO4 , K is +1
and the equation balanced as shown in Box 11.5.
Box 11.5 How to balance redox equations from partial ionic equations using the ion–
electron method
Example: You are to produce a balanced equation from 2. Balance the oxygen atoms on each side of each
the partial ionic equations deduced in Box 11.4. equation.
1. Balance the atom which changes oxidation number (a) If the reaction occurs in acid or neutral solution,
in each partial ionic equation: for each O atom deficient, add one molecule H2 O
MnO4- S Mn2 + no change necessary in to the side deficient.
either equation since Mn and
O are balanced
H 2 O2 S O2 on each side of the equation

(b) If the reaction occurs in alkaline solution, for Therefore multiply top equation by 2 and bottom
each O deficient add two OH - to the side deficient equation by 5:
and one molecule of H2 O to the other.
10e - + 16H + + 2MnO4- S 2Mn2 + + 8H2 O
Your reaction occurs in acid solution, so: 5H2 O2 S 5O2 + 10H + + 10e -
MnO4- S Mn2 + + 4H2 O
H 2 O2 S O2 6. Add the equations together:
10e - + 16H + + 5H2 O2 + 2MnO4-
3. Balance the hydrogen by addition of H + to the side S 2Mn2+ + 5O2 + 10H + + 8H2O + 10e-
deficient:
7. Cancel terms on opposite sides of the new equation:
8H + + MnO4- S Mn2 + + 4H2 O
H2 O2 S O2 + 2H + 6H + + 5H2 O2 + 2MnO4- S 2Mn2 + + 5O2 + 8H2 O
4. Balance the charge on each side of the equation by This ionic equation is sufficient to work out the mole ratio
the addition of electrons, each electron having a of the reacting species, i.e. 5 moles of H2 O2 will react
charge of - 1: with 2 moles of MnO4-. All the other ions, K+ and SO2- 4 ,
remain in solution and are unchanged by the reaction. If
5e - + 8H + + MnO4- S Mn2 + + 4H2 O the fully balanced equation is required, the ions can be
H2 O2 S O2 + 2H + + 2e - added to the ionic equation at the end of the process and
the numbers adjusted:
5. Balance the electrons in each equation, since num- 3H2SO4 + 5H2O2 + 2KMnO4
ber of electrons gained by oxidising agent must S 2MnSO4 + 5O2 + K2SO4 + 8H2O
equal number of electrons lost by reducing agent.
Text references
Haynes, W.M. (ed.) (2015) CRC Handbook of Chemistry Robinson, R.A. and Stokes, R.H. (2012) Electrolyte Solu-
and Physics, 96th edn. CRC Press, Boca Raton. tions. 2nd edn. Dover Publications, New York.

Burrows, A., Holman, J., Parsons, A., Pilling, G. and Price, O’Neil, M.J., Smith, A. and Henkelmann, P.E. (2013) The
G. (2013) Chemistry3: Inorganic, Organic and Physical Merck Index: An Encyclopaedia of Chemicals, Drugs
Chemistry, 2nd edn. Oxford University Press. and Biologicals, 15th edn. Merck & Co. Inc., White-
house Station.
Mendham, J., Denney, R.C., Barnes, J.D. and Thomas,
M.J.K. (2000) Vogel’s Textbook of Quantitative Chemi-
Study exercises
11.1 Practise calculations involving molar concentra- (a) 1000.00 mL of 1.0 M sodium chloride solution?
tions (see also study exercises 9.1 and 9.2). What (b) 250.00 mL of 0.2004 M KMnO4 solution?
mass of substance would be required to prepare (c) 400 mL of sodium hydroxide at 5% w/v?
each of the following aqueous solutions (answer (d) 300 mL of potassium nitrate at 10% w/w?
in grams, to four decimal places in each case):
(continued)

Study exercises (continued)
11.2 Practise expressing concentrations in differ- 11.3 Using the concepts of ‘oxidation numbers’ and
ent ways. Express all answers to four decimal ‘partial ionic equations’, balance the follow-
places. ing redox reactions which all take place in acid
solution.
(a) What is 4.000 g L-1 sodium hydroxide
expressed in terms of molarity? (a) K2Cr2O7 + FeSO4 + H2SO4
(b) What is 0.1 mol L-1 K2 Cr2 O7 expressed in S Cr2(SO4)3 + Fe2(SO4)3 + K2SO4 + H2O
g L-1 ? (b) Na2S2O3 + I2 S Na2S4O6 + NaI
(c) What is 5% v/v ethanol, expressed in (c) KIO3 + KI + H2SO4 S K2SO4 + I2 + H2O
terms of molarity? (Density of ethanol at (d) (COOH)2 + KMnO4 + H2SO4
25 °C = 0.7892 g mL-1 ). S K2SO4 + MnSO4 + CO2 + H2O
(d) What is 150 mmol L-1 glucose expressed in
terms of per cent concentration (% w/v)?

12 pH and buffer solutions
pH is a measure of the amount of hydrogen ions (H + ) in a solution. It is usual

Definitions to think of aqueous solutions as containing H + ions (protons), though protons
actually exist in their hydrated form, as hydronium ions (H3 O+ ). The proton
Acid – a compound that acts as a pro-
ton donor in aqueous solution.
concentration of an aqueous solution [H + ] is affected by several factors:
Base – a compound that acts as a pro- ●● Ionisation (dissociation) of water, which liberates protons and hydroxyl
ton acceptor in aqueous solution. ions in equal quantities, according to the reversible relationship:
Conjugate pair – an acid together with
its corresponding base. H2O ∆ H + + OH - [12.1]
Alkali – a compound that liberates
hydroxyl ions when it dissociates. Since ●● Dissociation of acids, according to the equation:
hydroxyl ions are strongly basic, this
will reduce the proton concentration. H -A ∆ H + + A- [12.2]
Ampholyte – a compound that can where H -A represents the acid and A- is the corresponding conjugate
act as both an acid and a base. Water base. The dissociation of an acid in water will increase the amount of pro-
is an ampholyte since it may dissoci- tons, reducing the amount of hydroxyl ions as water molecules are formed
ate to give a proton and a hydroxyl ion (eqn [12.1]). The addition of a base (usually, as its salt) to water will decrease
(amphoteric behaviour). the amount of H + , owing to the formation of the conjugate acid (eqn [12.2]).
●● Dissociation of alkalis, according to the relationship:
Safety Note Safe working with X+ OH - ∆ X+ + OH - [12.3]

strong acids or alkalis-these can be
highly corrosive; rinse with plenty of where X+ OH - represents the undissociated alkali. Since the dissociation
water if spilled. of water is reversible (eqn [12.1]), in an aqueous solution the production of
hydroxyl ions will effectively act to ‘mop up’ protons, lowering the proton
concentration.
Many compounds act as acids, bases or alkalis: those which are almost
completely ionised in solution are usually called strong acids or bases, while
weak acids or bases are only slightly ionised in solution (p. 102).
In an aqueous solution, most of the water molecules are not ionised. In
fact, the extent of ionisation of pure water is constant at any given temperature
and is usually expressed in terms of the ion product (or ionisation constant)
of water, Kw:
Kw = [H + ][OH - ][12.4]
Example The pH of 0.02 mol L-1 where [H + ] and [OH - ] represent the molar concentration (strictly, the activity)
HCl can be calculated as follows: of protons and hydroxyl ions in solution, expressed as mol L-1. At 25 °C, the ion
HCl is a strong acid giving product of pure water is 10-14 mol2 L-2 (i.e. 10-8 mol2 m -6). This means that
[H +] = [ Cl -] = 0.02 mol L-1. Therefore the concentration of protons in solution will be 10-7 mol L-1 (10-4 mol m -3),
pH = - log10(0.02) = 1.7. with an equivalent concentration of hydroxyl ions (eqn [12.1]). Since these
values are very low and involve negative powers of 10, it is customary to use
the pH scale, where:
Example The pH of a solution is pH = -log10 [H + ][12.5]

6.4. Therefore the [H +] = 10-pH,
i.e. 3.98 * 10-7 mol L-1. and [H + ] is the proton activity (see p. 104).

pH and buffer solutions
Table 12.1 Effects of temperature on the

ion product of water (Kw), H + ion concentra- Key Point While pH is strictly the negative logarithm (to the
tion and pH at neutrality. Values calculated base 10) of H + activity, in practice H + concentration in mol L -1
from Haynes (2015) (equivalent to kmol m -3 in SI terminology) is most often used in
place of activity, since the two are virtually the same, given the
[H+ ] at limited dissociation of H2O. The pH scale is not SI: nevertheless,
Temp. neutrality pH at it continues to be used widely in chemistry.
(°C) KW (mol2 L −2) (nmol L -1) neutrality
0 0.11 * 10 -4 33.9 7.47

The value where an equal amount of H + and OH - ions are present is
4 0.17 * 10 -4 40.7 7.39
termed neutrality: at 25 °C the pH of pure water at neutrality is 27.0. At this
10 0.29 * 10 -4 53.7 7.27 temperature, pH values below 7 are acidic while values above 7 are alkaline.
20 83.2 7.08 However, the pH of a neutral solution changes with temperature (Table 12.1),
0.68 * 10 -4
owing to the enhanced dissociation of water with increasing temperature. This
25 1.01 * 10 -4 100.4 7.00
must be taken into account when measuring the pH of any solution and when
30 1.47 * 10 -4 120.2 6.92 interpreting your results.
37 -4 154.9 6.81
Always remember that the pH scale is a logarithmic one, not a linear one:
2.39 * 10
a solution with a pH of 3.0 is not twice as acidic as a solution of pH 6.0, but
45 4.02 * 10 -4 199.5 6.70 a thousand times as acidic (i.e. contains 1000 times the amount of H + ions).
Therefore, you may need to convert pH values into proton concentrations
before you carry out mathematical manipulations (see Box 53.2). For similar
reasons, it is important that pH change is expressed in terms of the original and
final pH values, rather than simply quoting the difference between the values:
a pH change of 0.1 has little meaning unless the initial or final pH is known.
Table 12.2 Properties of some pH indicator
dyes Measuring pH
Dye Acid-base Useful pH electrodes
colour pH range
Accurate pH measurements can be made using a pH electrode, coupled to a
change
pH meter. The pH electrode is usually a combination electrode, comprising
Thymol blue Red-yellow 1.2–6.8 two separate systems: an H + @sensitive glass electrode and a reference elec-
(acid) trode which is unaffected by H + ion concentration (see Fig. 12.1). When this
Bromophenol Yellow-blue 1.2–6.8 is immersed in a solution, a pH-dependent voltage between the two electrodes
blue can be measured using a potentiometer. In most cases, the pH electrode assem-
Methyl orange Red-yellow 2.8–4.0 bly (containing the glass and reference electrodes) is connected to a separate
Congo red Blue-red 3.0–5.2 pH meter by a cable, although some hand-held instruments (pH probes) have
combination electrodes and meter within the same assembly, often using an
Bromocresol Yellow-blue 3.8–5.4
green
H + @sensitive field effect transistor in place of a glass electrode, to improve
durability and portability.
Methyl red Red-yellow 4.3–6.1
Box 12.1 gives details of the steps involved in making a pH measurement
Litmus Red-blue 4.5–8.3 with a glass pH electrode and meter.
Chlorophenol red Yellow-red 4.8–6.3
Bromocresol Yellow-purple 5.2–6.8 pH indicator dyes
purple These compounds (usually weak acids) change colour in a pH-dependent man-
Bromothymol Yellow-blue 6.0–7.6 ner. They may be added in small amounts to a solution, or they can be used in
blue paper strip form. Each indicator dye usually changes colour over a restricted pH
Neutral red Red-yellow 6.8–8.0 range (Table 12.2): universal indicator dyes/papers make use of a combination
Phenol red Yellow-red 6.8–8.2 of individual dyes to measure a wider pH range. Dyes are not suitable for accu-
rate pH measurement as they are affected by other components of the solution
1-Naphthol- Yellow-blue 7.2–8.6
phthalein
including oxidising and reducing agents and salts. However, they are useful for:
Phenol- None-red 8.3–10.0 ●● estimating the approximate pH of a solution;
phthalein
●● determining a change in pH, e.g. at the end-point of a titration.

Box 12.1 Using a glass pH electrode and meter to measure the pH of a solution
The following procedure should be used whenever 4. Rinse the electrode assembly with distilled water
you make a pH measurement: consult the manufac- and gently dab off the excess water onto a clean tis-
turer’s handbook for specific information, where nec- sue: check for visible damage or contamination of the
essary. Do not be tempted to miss out any of the steps glass electrode (consult a member of staff if the glass
detailed below, particularly those relating to the effects is broken or dirty). Also check that the solution within
of temperature, or your measurements are likely to be the glass assembly is covering the metal electrode.
inaccurate.
5. Calibrate the instrument: set the meter to ‘pH’ mode,
if appropriate, and then place the electrode assembly in
pH meter a standard solution of known pH, usually pH 7.00. This
(potentiometer)
solution may be supplied as a liquid, or may be pre-
pared by dissolving a measured amount of a calibra-
tion standard in water: calibration standards are often
provided in tablet form, to be dissolved in water to give
silver–silver chloride
a particular volume of solution. Adjust the calibration
electrode control to give the correct reading. Remember that your
calibration standards will only give the specified pH at
a particular temperature, usually either 20 °C or 25 °C.
saturated If you are working at a different temperature, you must
KCI establish the actual pH of your calibration standards,
solution
either from the supplier or from literature information.
calomel reference
porous electrode
plug
6. Remove the electrode assembly from the calibration
H+-sensitive
solution and rinse again with distilled water: dab off
glass electrode the excess water. Basic instruments have no further cali-
HCI
solution bration steps (single-point calibration), while the more
refined pH meters have additional calibration procedures.
If you are using a basic instrument, you should
test solution
check that your apparatus is accurate over the appro-
Fig. 12.1 Measurement of pH using a combination pH priate pH range by measuring the pH of another
electrode and meter. The electrical potential difference standard whose pH is close to that expected for
recorded by the potentiometer is directly proportional to the test solution. If the standard does not give the
the pH of the test solution. expected reading, the instrument is not functioning
correctly: consult a member of staff.
If you are using an instrument with a slope control
1. Stir the test solution thoroughly before you make function, this will allow you to correct for any deviation
any measurement: it is often best to use a magnetic in electrical potential from that predicted by the the-
stirrer. Leave the solution for sufficient time to allow oretical relationship (at 25 °C, a change in pH of 1.00
equilibration at lab temperature. unit should result in a change in electrical potential
2. Record the temperature of every solution you use, of 59.16 mV) by performing a two-point calibration.
including all calibration standards and samples, since Having calibrated the instrument at pH 7.00, immerse
this will affect Kw, neutrality and pH. in a second standard at the same temperature as that
of the first standard, usually buffered to either pH 4.00
3. Set the temperature compensator on the meter to or pH 9.00, depending upon the expected pH of your
the appropriate value. This control makes an allow- samples. Adjust the slope control until the exact value
ance for the effect of temperature on the electrical of the second standard is achieved (Fig. 12.2). A pH
potential difference recorded by the meter: it does electrode and meter calibrated using the two-point
not allow for the other temperature-dependent effects method will give accurate readings over the pH range
mentioned elsewhere. Basic instruments have no from 3 to 11: laboratory pH electrodes are not accurate
temperature compensator, and should only be used outside this range, since the theoretical relationship
at a specified temperature, either 20 °C or 25 °C, oth- between electrical potential and pH is no longer valid.
erwise they will not give an accurate measurement.
More sophisticated systems have automatic temper- 7. Once the instrument is calibrated, measure the pH
ature compensation. of your solution(s), making sure that the electrode
(continued)

Box 12.1 (continued)
stabilise in each solution before taking a measure-

+300
ment: for unbuffered solutions, this may take several
2 minutes, so do not take inaccurate pH readings due
+200
to impatience!
1
+100 8. After use, the electrode assembly must not be
allowed to dry out. Most pH electrodes should be
pH scale stored in a neutral solution of KCl, either by suspend-
3 4 5 6 8 9 10 11
ing the assembly in a small beaker, or by using an
–100
electrode cap filled with the appropriate solution (typ-
ically 1.0 mol L-1 KCl buffered at pH 7.0). However,
many labs simply use distilled water as a storage
–200
solution, leading to loss of ions from the interior of
the electrode assembly. In practice, this means that
–300
electrical pH electrodes stored in distilled water will take far
potential longer to give a stable reading than those stored in
(mV) KCl.
Fig. 12.2 The relationship between electrical potential 9. Switch the meter to zero (where appropriate), but do
and pH. The solid line shows the response of a calibrated not turn off the power: pH meters give more stable
electrode while the other plots are for instruments requir- readings if they are left on during normal working
ing calibration: 1 has the correct slope but incorrect iso- hours.
potential point (calibration control adjustment is needed); Problems (and solutions) include: inaccurate and/or
2 has the correct isopotential point but incorrect slope unstable pH readings caused by crosscontamination
(slope control adjustment is needed). (rinse electrode assembly with distilled water and blot
dry between measurements); development of a protein
assembly is rinsed thoroughly between measure- film on the surface of the electrode (soak in 1% w/v pep-
ments. You should be particularly aware of this sin in 0.1 mol L-1 HCl for at least an hour); deposition
requirement if your solutions contain organic bio- of organic or inorganic contaminants on the glass bulb
logical material, e.g. soil, tissue fluids, protein solu- (use an organic solvent, such as acetone, or a solution
tions, since these may adhere to the glass electrode of 0.1 mol L-1 disodium ethylenediaminetetraacetic acid,
and affect the calibration of your instrument. If your respectively); drying out of the internal reference solu-
electrode becomes contaminated during use, check tions (drain, flush and refill with fresh solution, then allow
with a member of staff before cleaning: avoid touch- to equilibrate in 0.1 mol L-1 HCl for at least an hour);
ing the surface of the glass electrode with abrasive cracks or chips to the surface of the glass bulb (use a
material. Allow sufficient time for the pH reading to replacement electrode).
Buffers
Safety Note Preparing a dilute
acid solution using concentrated Rather than simply measuring the pH of a solution, you may wish to control
acid-always slowly add the concen- the pH, during EDTA complexation titrations (see p. 203) or preparative exper-
trated acid to water, not the reverse, iments involving carbonyl compounds, and one of the most effective ways to
since the strongly exothermic process control pH is to use a buffer solution.
can lead to a violent reaction with water. A buffer solution is usually a mixture of a weak acid and its conjugate
base. Added protons will be neutralised by the anionic base while a reduction
in protons, e.g. due to the addition of hydroxyl ions, will be counterbalanced by
Safety Note Preparing an alkali
dissociation of the acid (eqn [12.2]); thus the conjugate pair acts as a ‘buffer’
solution-typically, the alkali will be in
to pH change.
solid form (e.g. NaOH) and addition
The British standard for the pH scale is an aqueous solution of potassium
to water will rapidly raise the temper-
hydrogen phthalate (0.05 M), which has a pH of 4.001 at 20 °C and is often
ature of the solution: use only heat-
used as a calibration solution for pH meters.
resistant glassware, cooled with water,
if necessary.

Buffer capacity and the effects of pH

Safety Note If you spill concen-
trated H2SO4 on your skin it is neces- The extent of resistance to pH change is called the buffer capacity of a solu-
sary to first wipe your skin with a dry tion. The buffer capacity is measured experimentally at a particular pH by
cloth or tissue. Then, wash the affected titration against a strong acid or alkali: the resultant curve will be strongly
area with copious amounts of water. sigmoidal, with a plateau where the buffer capacity is greatest (Fig. 12.3).
Dispose the cloth/tissue after dousing The mid-point of the plateau represents the pH where equal quantities of acid
in water. and conjugate base are present, and is given the symbol pKa, which refers
NOTE: By trying to remove H2SO4 first to the negative logarithm (to the base 10) of the acid dissociation constant,
produces heat which will burn the skin. Ka, where
[H + ][A- ]
Ka = [12.6]
[HA]
Definition
By re-arranging eqn [12.6] and taking negative logarithms, we obtain:
Buffer solution – one which resists a
change in H + concentration (pH) on [A- ]
pH = pKa + log10 [12.7]
addition of acid or alkali. [HA]
This relationship is known as the Henderson-Hasselbalch equation and it

Potassium hydrogen phthalate (KHP) shows that the pH will be equal to the pKa when the ratio of conjugate base
is the mono potassium salt of a weak to acid is unity, since the final term will be zero. Consequently, the pKa of a
organic dibasic acid. buffer solution is an important factor in determining the buffer capacity at a
particular pH. In practical terms, this means that a buffer solution will work
COO-K+
most effectively at pH values about one unit either side of the pKa.
COOH
Selecting an appropriate buffer
When selecting a buffer, you should be aware of certain limitations to its use.
Citric acid and phosphate buffers readily form insoluble complexes with diva-
lent cations, while phosphate can also act as a substrate, activator or inhibitor
of certain enzymes. Both of these buffers contain biologically significant quan-
tities of cations, e.g. Na+ or K+ . TRIS (Table 12.3) is often toxic to biological
systems: owing to its high lipid solubility it can penetrate membranes, uncou-
pling electron transport reactions in whole cells and isolated organelles. In
addition, it is markedly affected by temperature, with a 10-fold increase in H +
concentration from 4 °C to 37 °C. A number of zwitterionic molecules (pos-
effective sessing both positive and negative groups) have been introduced to overcome
buffering range some of the disadvantages of the more traditional buffers. These newer com-
pounds are often referred to as ‘Good buffers’, to acknowledge the early work
pH or buffer capacity
pKa of Dr N.E. Good and co-workers: HEPES is one of the most useful zwitterionic
buffers, with a pKa of 27.5 at 25 °C.
These zwitterionic substances are usually added to water as the free acid:
the solution must then be adjusted to the correct pH with a strong alkali, usually
pH
NaOH or KOH. Alternatively, they may be used as their sodium or potassium
buffer capacity
salts, adjusted to the correct pH with a strong acid, e.g. HCl. Consequently, you
may need to consider what effects such changes in ion concentration may have
in a solution where zwitterions are used as buffers.
Fig. 12.4 shows a number of traditional and zwitterionic buffers and their
alkali added
effective pH ranges. When selecting one of these buffers, aim for a pKa which
Fig. 12.3 Theoretical pH titration curve for is in the direction of the expected pH change (Tables 12.2 and 12.3). For exam-
a buffer solution. pH change is lowest and ple, HEPES buffer would be a better choice of buffer than PIPES for use at pH
buffer capacity is greatest at the pKa of the 7.2 for experimental systems where a pH increase is anticipated, while PIPES
buffer solution. would be a better choice for where acidification is expected.

Table 12.3 pKa values at 25 °C of some acids

and bases (upper section) and some large phosphate phosphate
organic zwitterions (lower section) com-

monly used in buffer solutions. For poly- citrate TRICINE
protic acids, where more than one proton
may dissociate, the pKa values are given for succinate CHES
each ionisation step. Only the trivial acro-
nyms of the larger molecules are provided: acetate HEPES borate
their full names can be obtained from the

catalogues of most chemical suppliers MES TRIS CAPS
Acid or base pKa value(s) PIPES
Acetic acid 4.8 1 2 3 4 5 6 7 8 9 10 11

pH
Carbonic acid 6.1, 10.2
Citric acid 3.1, 4.8, 5.4 Fig. 12.4 Useful pH ranges of some commonly used buffers.
Glycylglycine 3.1, 8.2
Phthalic acid 2.9, 5.5 Preparation of buffer solutions
Phosphoric acid 2.1, 7.1, 12.3 Having selected an appropriate buffer, you will need to make up your solution
to give the desired pH. You will need to consider two factors:
Succinic acid 4.2, 5.6
TRIS* 8.3 1. The ratio of acid and conjugate base required to give the correct pH.
Boric acid 9.2 2. The amount of buffering required; buffer capacity depends upon the abso-
lute quantities of acid and base, as well as their relative proportions.
MES 6.1
PIPES 6.8 In most instances, buffer solutions are prepared to contain between 10 mmol L-1
and 200 mmol L-1 of the conjugate pair. While it is possible to calculate the
MOPS 7.2
quantities required from first principles using the Henderson-Hasselbalch
HEPES 7.5
equation, there are several sources which tabulate the amount of substance
TRICINE 8.1 required to give a particular volume of solution with a specific pH value for
TAPS 8.4 a wide range of traditional buffers (e.g. Perrin and Dempsey, 1974). For tra-
CHES 9.3
ditional buffers, it is customary to mix stock solutions of acidic and basic
components in the correct proportions to give the required pH (Table 12.4).
CAPS 10.4
For zwitterionic acids, the usual procedure is to add the compound to water,
*Note that this compound is hygroscopic and
should be stored in a desiccator.
Table 12.4 Preparation of sodium phosphate buffer solutions for use at
25 °C. Prepare separate stock solutions of (a) disodium hydrogen phos-
phate and (b) sodium dihydrogen phosphate, both at 0.2 mol L-1. Buffer
solutions (at 0.1 mol L-1.) are then prepared at the required pH by mixing
together the volume of each stock solution shown in the table, and then
diluting to a final volume of 100 mL using distilled or deionised water
Volume of stock (a) Volume of stock (b)

Required pH (at 25 °C) Na2HPo4 (mL) NaH2Po4 (mL)
6.0 6.2 43.8

6.2 9.3 40.7
6.4 13.3 36.7
6.6 18.8 31.2
6.8 24.5 25.5
7.0 30.5 19.5
7.2 36.0 14.0
7.4 40.5 9.5

Volume of stock (a) Volume of stock (b)

Required pH (at 25 °C) Na2HPo4 (mL) NaH2Po4 (mL)
7.6 43.5 6.5

7.8 45.8 4.2
8.0 47.4 2.6
and then bring the solution to the required pH by adding a specific amount of
strong alkali or acid (obtained from tables). Alternatively, the required pH can
be obtained by dropwise addition of alkali or acid, using a meter to check the
pH, until the correct value is reached. When preparing solutions of zwitterionic
buffers, the acid may be relatively insoluble. Do not wait for it to dissolve fully
before adding alkali to change the pH – the addition of alkali will help bring
the acid into solution (but make sure it has all dissolved before the desired pH
is reached).
Finally, when preparing a buffer solution based on tabulated information,
always confirm the pH with a pH meter before use.
Text references
Haynes, W.M. (ed.) (2015) CRC Handbook of C
hemistry Perrin, D.D. and Dempsey, B. (2012) Buffers for pH and
and Physics, 96th edn. CRC Press, Boca Raton. Metal Ion Control. Springer Science and Business
Media, London.

Butler, J.N. (1998) Ionic Equilibrium: Solubility Galster, H. (1991) pH Measurement: Fundamentals, Meth-
and pH calculations. John Wiley & Sons Ltd ods, Applications, Instrumentation. John Wiley & Sons
Chichester. Ltd., New York.
Clark, J. (2000) Calculations in AS/A Level Chemistry. Rilbe, H. (1996) pH and Buffer Theory – a New Approach.
Longman, Harlow, Essex. John Wiley & Sons Ltd., New York.
Study exercises
12.1 Practise interconverting pH values and proton 12.2 Practise using the Henderson-Hasselbalch equa-
concentrations. Express all values to four sig- tion. What are the relative proportions of the
nificant figures. deprotonated (A -) and protonated (HA) forms of
each substance at the following pH values:
(a) What is a pH 7.4 expressed as [H +] in mol L-1?
(b) What is a pH 4.1 expressed as [H +] in (a) ethanoic acid (pKa = 4.8) for use in an exper-
mol L-1? iment at pH 3.8;
(c) What is the pH of a solution of [H +] at (b) boric acid (pKa = 9.2) for use in an experi-
2 * 10-5 mol L-1? ment at pH 9.5;
(d) What is the pH of a solution of [H +] at (a) HEPES (pKa = 7.5) for use in an experiment
2 * 10-12.5 mol L-1? at pH 8.1?

Laboratory techniques
13. Melting points 123
14. Recrystallisation 128
15. Solvent extraction 139
16. Distillation 145
17. Reflux 155
18. Evaporation 161
19. Inert atmosphere methods 166
20. Combinatorial chemistry 173

13 Melting points
Melting points are measured (determined) for four reasons:

Purity of solids – melting points do
not vary significantly with changes in 1. The melting range and upper limit are an indication of the purity of the
atmospheric pressure, whereas the sample.
boiling points of pure liquids are not 2. Comparison of the melting point with the literature may indicate the iden-
constant: they vary with changes in tity of the compound or confirm that it is not the compound required.
atmospheric pressure and cannot be
used as a measure of purity. 3. If the compound is new, other scientists will need the information.
4. The compound can be identified with reasonable certainty by taking a
mixed melting point (p. 127).
Criterion of purity
Melting point depression – only impu-

Pure solid covalent organic compounds and many inorganic complexes incor-
rities which will dissolve in the com-
porating organic ligands have definite melting points. The pure solid will melt
pound will lower its melting point, e.g.
reproducibly over a narrow temperature range, usually less than 1 °C, and this
reactants, by-products, solvents. Insol-
melting range is known as the melting point. If the compound is not pure, the
uble impurities, e.g. salts, charcoal, fil-
melting range will increase significantly and the upper end of the melting range
ter paper, grit, will not dissolve in the
will be lowered. Thus the melting point (m.pt.) of a compound is a measure
melted compound.
of its purity (p. 126). Other methods are used routinely to estimate purity of
solid compounds, such as NMR (p. 334), and the presence of a single ‘spot’ or
a single peak on a chromatogram from thin-layer (p. 260), gas–liquid (p. 255)
or high-performance liquid (p. 265) chromatography, but melting point remains
When you synthesise soild compounds the standard measure of purity.
in the laboratory – you will always be
asked to determine their melting points
so that your practical expertise can be Key Point The term melting point really means the melting
assessed. range of a chemical and in your laboratory report you should
always quote the measured melting range under the heading
‘melting point’.
Melting point apparatus

The equipment for measuring the melting point of a solid varies in complex-
ity from a simple oil bath heated with a microburner to a microscope with a
heated stage as shown in Fig. 13.1. The essential components of a melting
point apparatus are:
●● A sample holder: usually a glass capillary tube sealed at one end in the case
Thermometers – make sure that you of the oil bath and heated block systems, or a pair of microscope slides on
use a partial-immersion thermometer an electrically heated plate in the Kofler block.
not a total-immersion thermometer. ●● A temperature recording device, placed as near to the sample as possible.
The type is written on the back of the Usually this is a thermometer but it can be a thermocouple probe with digital
thermometer. readout.
●● A heat source with fine control to allow a gradual increase in temperature.
These sources vary with the sophistication of the equipment.
In the undergraduate laboratory you are likely to use only the simpler
systems such as the oil bath or heated block apparatus.

Melting points
rubber
band
thermometer
capillary
tube
capillary
tube
(b)
(a)
capillary tube
thermometer
(c) (d)
Fig. 13.1 Melting point apparatus: (a) oil bath; (b) heated block – ther-
mometer readout; (c) heated block – digital readout; (d) Kofler hot-stage
microscope.
Capillary tubes
Breaking capillary tubes – if the capil-
lary tube is sealed at both ends, you can Capillary tubes for melting point measurement are available commercially and
break it into two half-sized single-end- are supplied open at both ends or closed at one end or closed at both ends. To
sealed melting point tubes by scoring seal one end of an open capillary tube, just touch the end of the capillary tube
the mid-point of the tube with a glass onto the outer ‘layer’ of the hot flame of a microburner (see Fig. 13.2). The end
file and then snapping the tube at the of the tube will collapse in and seal the tube. Make sure that the tube is sealed,
score mark. If you don’t score the glass, i.e. there is not a fine line in the sealed end, and that there is no large globule of
the edges of the break will be uneven glass on the end of the tube, otherwise it may not fit into the hole in the heating
and the tube will be difficult to fill. block of the melting point apparatus. Similarly, if you push the tube too far
into the flame, the tube will bend and therefore not fit into the heating block.
124 Laboratory techniques

Melting points
To put the compound into the capillary tube, place a little of the dry com-
pound in a small heap on a watch-glass and press the open end of the tube into
the heap, trapping a plug of chemical in the opening. The chemical can be
WRONG!
moved to the sealed end by turning the tube over and tapping it on the bench,
Tube has been or by vibrating it by rubbing it against the thread of the screw on a clamp, or
overheated dropping it down a long glass tube onto the laboratory bench. Remember: you
tube tilted to stages for
prevent water correct sealing only need a 2–3 mm length of sample in the bottom of the capillary tube.
condensation inside
Fig. 13.2 Sealing a melting point capillary

Oil bath apparatus
tube. The component parts of a typical oil bath melting point apparatus are shown
in Fig. 13.3.
●● Check that the mineral oil is clean and contains no water (p. 90) and that the
thermometer bulb is only two-thirds full, to allow for expansion. Clamp the oil bath to a
cork
support stand (p. 82).
split for
viewing rubber
●● Check that your thermometer is of the appropriate range, that the mercury
m.pt.
thermometer band
tube
thread is intact and the glass, in particular the bulb, is not cracked.
liquid sample ●● Attach the capillary to the thermometer with a rubber ring, making sure that
paraffin size
bath 2–3 mm the compound is next to the thermometer bulb and remember to hold the
thermometer near the bulb while attaching the capillary tube (p. 68).
microburner
●● Press the thermometer into the ‘notched’ cork, making sure that you can see
the thermometer scale, and trial fit the thermometer, sample and cork into
Fig. 13.3 Components of an oil bath melt- the oil bath making sure that the thermometer bulb and sample are in the
ing point apparatus. centre of the oil bath and that the rubber band is not in the oil, and will not
be covered by oil, when the oil expands on heating.
●● If adjustment is needed, carefully slide the cork up or down the thermometer
If the rubber band comes into contact
(p. 68).
with the oil it will expand – ºthe capillary
tube will drop off into the oil bath and it ●● You can attach a melting point capillary to both sides of the thermometer
will discolour the oil so that you will not bulb to carry out two separate melting point determinations simultaneously.
be able to see when your sample melts.
Electric heated block apparatus
The heating block usually has three holes, to permit three simultaneous meas-
Safety Note The ‘notch’ in the cork urements of melting points. Always ensure that:
is essential: it allows the thermometer
●● The heating block is at room temperature at the outset.
to be read over all its length, it allows
the thermometer to be gripped by the ●● The light in the heating block works.
cork and most importantly, it allows the
●● The thermometer is undamaged (as above) and fits snugly into its hole in
heated air to escape from the appara-
the heating block.
tus. Never use a cork without the ‘notch’.
●● All heating controls are set at zero.
●● The capillary tubes slide easily into their holes in the heating block.
Safety Note Mercury vapour is The usual problems encountered with this equipment are broken capillary
a severe cumulative and chronic haz- tubes or broken thermometer bulbs in the heating block. Consult your instructor
ard and the normal vapour pressure of in these cases.
mercury, at room temperature, is many
times above the control limit (CL) of
0.05 mg m -3 If you break a thermom- Melting point determination
eter or find or suspect the presence The general procedure is to heat the oil bath with the microburner using the
of mercury, inform your instructor technique described on p. 88, or to use the heating control to give a temperature
immediately. rise of about 10 °C per minute. When the temperature is about 20 °C below

Melting points
the melting point, the rate of heating must be reduced to about 2 °C per minute
and continued at this rate until the compound has melted. The melting point is
the temperature range from where the first drop of liquid appears to where the
last crystal dissolves into the liquid.
Key Points
●● The most common error is heating the sample too quickly.
There is often a lag between slowing the heating rate and the
reduction in temperature increase, resulting in a reading of
Heating too fast – if you record a melt- the melting point which is too high.
ing point which is higher than the liter- ●● If you do not know what the compound is, you will not know
ature value, you are implying that your its expected melting point. Therefore, you should deter-
compound has a purity 7100%, which mine an approximate value and then repeat the procedure
is impossible. Therefore it must be a dif- to give an accurate measurement, by reducing the heating
ferent compound to the one expected. rate, particularly for the final stage near the melting point.
The boost heater on the electrical apparatus can be used for
finding the approximate melting point but not for accurate
determination.
●● If you are carrying out several melting point measurements, it
is common sense to measure the melting point of the lowest

melting compound first – less time for the apparatus to cool
to room temperature.
●● If you miss the melting point, do not allow the sample to solid-
ify and then retake the melting point with the same sample:
some decomposition may have occurred on melting.
●● It is normal practice to make at least two measurements of
the melting point, one approximate and at least one accurate

reading.
Thermometer calibration
Calibrated thermometers – if your Your thermometer may be a major source of error in melting point measure-
compound is only slightly impure and ment. Occasionally thermometers for routine laboratory use may not be accu-
has a melting point 2 °C lower than the rate and may read up to 10 °C high or low. To avoid this problem you should
literature value, a thermometer reading always calibrate your thermometer to determine any error and be able to correct
10 °C low will give you an error of 12 °C, for it.
indicating a low level of purity and To calibrate your thermometer you must measure the melting points of
hence a low grade or false information a series of very pure compounds, available commercially, having a range of
to fellow scientists. melting points similar to the range over which you will use the thermometer;
a general-purpose series is shown in Table 13.1. Having measured the melting
points of each of the pure compounds, take the mid-point of each value for
the thermometer reading of melting point for each compound and the mid-
Table 13.1 A typical series of standards for point of the literature melting point of each compound and plot them graphi-
thermometer calibration cally using literature temperature as the y-axis and the thermometer reading as
the x-axis. The straight-line plot will obey the equation y = mx + c, where
Compound Literature m.pt.
y = real temperature (literature m.pt.), x = thermometer reading, m = slope
Naphthalene 79–80 °C and c = intercept. If you use a computer and suitable program, the values of
Benzoic acid 121 °C m and c will be calculated and you then solve the equation for using the ther-
4-Nitroaniline 147 °C mometer reading (x-axis values) to find the real temperature of melting (y-axis
4-Toluic acid 180–182 °C
values) to find the true melting range.
Of course, if you break or lose your thermometer, you must calibrate
Anthracene 216–218 °C
another.

Melting points
Mixed melting point determination

You can confirm the identity of a compound by determining a mixed melting
point. If you prepare a mixture of your unknown chemical and the one you
suspect it may be and measure the melting point of the mixture, then there are
two possible results:
1. The melting point of the mixture is the same as the pure compound, which
means that the unknown compound and the known compound are the
same.
2. The melting point of the mixture is lower than either of the two pure com-
ponents and the melting range is large. This is because the two compounds
are different, with the result that one is an impurity in the other.
For example, both benzoic acid and mandelic acid are white crystalline
solids which melt at 121 °C. However a 1:1 mixture of the two compounds
begins to melt at about 80 °C.
The usefulness of mixed melting points is limited in that you must have
some idea of the chemical nature of your unknown compound and a sample of
the suspected compound must be available.

Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch- Lehman, J.W. (2009) Student Lab Companion: Laboratory
ell, A.R. (1991) Vogel’s Textbook of Practical Organic Techniques for Organic Chemistry, 2nd edn. Pearson,
Chemistry, 5th edn. Longman, Harlow, Essex. Harlow.
Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper- Zubrick, J.W. (2015) The Organic Chem Lab Survival
imental Organic Chemistry, 2nd edn. Blackwell Science Manual. A student’s guide to techniques, 10th edn. John
Ltd, Oxford. Wiley & Sons Ltd, Chichester.
Study exercises
13.1 You have prepared a solid crystalline compound 13.3 You have carried out a synthetic procedure
which you think is N-phenylethanamide (litera- which results in a mixture of 2-nitroaniline (liter-
ture m.pt. = 114 °C) and its m.pt. range is 108– ature m.pt = 92 °C) and 4-nitroaniline (literature
110 °C. Draw three possible conclusions from this m.pt. = 148°C). Purification by recrystallisation
result. is said to separate the two isomers and your
13.2 You have obtained a sample of a white crystal- sample, when mixed with pure 4-nitroaniline,
line compound which you think is benzoic acid has a m.pt. range 147–148 °C. What can you
(literature m.pt. = 121 °C) and its m.pt. range is conclude?
found to be 120–128, °C. Draw three possible con-
clusions from this result.

14 Recrystallisation
The products from many synthetic preparations are seldom pure and the tech-
Recrystallisation – involves allowing a
nique of recrystallisation, which involves dissolving the impure material in a
hot solution of the required compound
hot solvent and then cooling the solution to produce crystals, is routinely used
to cool.
to purify covalent organic and inorganic solids.
In general there are three types of impurities which are removed by the
Crystallisation – implies allowing the recrystallisation process:
solvent to evaporate from a solution of 1. Insoluble material: anti-bumping granules, pieces of filter paper, traces of
the compound. Crystallisation will not drying agents, grit, hair and other materials which may have been present
remove solvent-soluble impurities since in the starting chemicals.
they will be deposited as the solvent
evaporates.
2. Small quantities of unreacted starting chemicals and/or by-products from
side reactions or other isomers.
3. Very small amounts of coloured by-products resulting from oxidation or
polymerisation of the chemicals used.
Limits of purification – crude solids

Recrystallisation is designed to remove all these types of impurity and
containing only up to 10%–15% impuri-
provide a pure product suitable for melting point measurement. Purification
ties can be purified by recrystallisation.
by recrystallisation is based on the theory of saturated solutions (p. 106) and a
Otherwise chemical purification or chro-
suitable recrystallisation solvent is one in which the chemical to be purified is
matography (p. 263) will be required to
insoluble in the cold solvent and soluble in the hot solvent.
produce a compound, which can then
When the crude reaction product is dissolved in the hot solvent the insol-
be further purified by recrystallisation.
uble impurities (type 1 above) can be removed by hot filtration (p. 131). When
the hot solution is allowed to cool, the solution becomes saturated with the
desired compound and it precipitates from the cold solution. The cold solution
does not become saturated with the lower concentration of the contaminants of
type 2, which therefore remain in solution. Coloured impurities (type 3) can be
removed by absorption using charcoal as described on p. 133.
The recrystallisation process can be divided into three separate steps:
1. Selection of a suitable solvent.
2. Recrystallisation of the crude compound.
3. Drying the purified solid.
Key Point Practice in the technique of recrystallisation is

important, since the aim of the procedure is to produce the max-
imum quantity of the highest quality product. Poor technique
often results in low recovery of high-quality product or high
recovery of low-quality product.
Solvents for recrystallisation

You can decide on a suitable solvent for recrystallisation of your crude chem-
ical product in several ways:
●● The experimental protocol may tell you which solvent to use.
●● If you know the identity of the compound you have made, reference texts
may indicate a suitable solvent (e.g. Buckingham and Macdonald, 1995;
Haynes, 2015).

Recrystallisation
●● If you are not sure of the identity of the compound you have prepared or if
it is a new compound, you must carry out a ‘solvent selection’ to find out
which solvent is the most appropriate (p. 130).
When selecting a suitable solvent for recrystallisation, chemists work to
the general rule ‘like dissolves like’ when considering the polarity of the chem-
ical to be recrystallised and the polarity of the solvent. In general, solvents
for recrystallisation are classified in terms of polarity and miscibility. Solvent
polarity depends upon the overall distortion of the electron clouds in the cova-
lent bonds within the solvent molecules, resulting in a dipole. The greater the
dipole, the greater the polarity of the solvent, e.g. trichloromethane has three
polarised C-Cl bonds, giving an overall dipole to the molecule and producing
a good solvent for many molecules, but in tetrachloromethane all of the C-Cl
dipoles cancel out, giving a non-polar solvent with different solvent properties
in comparison. An additional feature that adds solvating power is hydrogen
bonding. Thus water, a polarised molecule, can form hydrogen bonds with
oxygen and nitrogen atoms in solutes, dissolving them efficiently.
Yields from recrystallisation – some
When you are choosing a solvent for recrystallisation, look for the follow-
of the compound that you require
ing general characteristics:
will remain dissolved in the solvent, ●● A high dissolving power for the solute at high temperature and a low dissolv-
because it has to form a saturated ing power at room temperature or below, so that a high recovery of purified
solution. Therefore you can never compound can be achieved.
recover 100% of your compound by
recrystallisation.
●● A high or negligible dissolving power for the impurities, so that they will
either be filtered off or remain in solution.
●● A relatively low boiling point, to facilitate drying the purified compound.
Definitions The miscibility of solvents with each other must be taken into account
when attempting mixed-solvent recrystallisations (see p. 130); the properties
Solvents which mix in all proportions,
of some common recrystallisation solvents, which you are likely to encounter
such as water and ethanol, are said to
in your laboratory work, are shown in Table 14.1. Remember that this is only a
be miscible, whereas solvents which
general list of solvents; further information on solvent properties can be found
do not mix, e.g. water and petrol, are
in standard textbooks such as Harwood et al. (2015), Loewenthal (1992) and
immiscible.
Furniss et al. (1991).
Table 14.1 Selected solvent properties
b.pt. Miscibility Comments: class of organic

Solvent (°C) Polarity with water compounds recrystallised
Water 100 V. high Yes Always use when suitable:

salts, aromatic acids
Ethanol 78 High Yes Flammable: alcohols, acids,
amides
Propanone 56 High Yes Flammable: carbonyl
compounds
Ethyl ethanoate 78 Medium No Flammable: esters
Dichloromethane 41 Medium No Toxic: halogen compounds,
ethers
Toluene 110 Low No Flammable: hydrocarbons
Light petroleum 40–60 V. low No Flammable: hydrocarbons

Recrystallisation
Solvent selection
To find a suitable solvent for recrystallisation you must carry out a series of
tests measuring the solubility of your crude compound in a series of solvents
(cold and hot) of varying solvent polarity. This series of tests is called ‘solvent
selection’ and is carried out on a test tube scale but, as your technique improves,
you can carry out the tests using semi-micro scale since you will use less of
your compound during the process. The procedure for solvent selection at the
test tube scale is described in Box 14.1 and modification of the procedure to
semi-micro scale requires only a corresponding reduction of the quantities of
solvent and solute used.
Key Point When carrying out the solvent selection experi-

ments you must always cool the solution after heating. Do not
assume that if the compound is insoluble in the cold solvent
and dissolves when heated, it will always precipitate on cooling.
Mixed solvents
When no single solvent is found to be suitable for recrystallisation, then a mixed
solvent system must be used. There are three essential properties required for a
pair of solvents to be used in a mixed-solvent system:
Using mixed solvents for recrystalli- 1. The two solvents must be miscible in all proportions over the temperature
sation – never assume that a mixed range to be used.
solvent system is a mixture of equal
volumes of the two solvents. The ratio 2. The solute must be insoluble in one of the solvents.
of the two solvents is established 3. The solute must be soluble in the other solvent.
practically during the recrystallisation
experiment. It is an advantage if the two solvents have similar boiling points within
20–30 °C.
Suitable common solvent pairs from the solvents given in Table 14.1 are
water/ethanol and dichloromethane/light petroleum (b.pt. 40–60 °C), but many
other combinations are possible. One of the most frequently encountered mixed
solvent systems is ‘aqueous ethanol’ (water/ethanol) in which the compound to
be recrystallised is insoluble in water and very soluble in ethanol.
You can identify the solubility characteristics of your compound using the
solvent selection procedure shown in Box 14.1 and modifications for mixed
solvent selection are given in Box 14.2.
The recrystallisation process

Having chosen a suitable solvent system, the process to be used to purify the
bulk of your impure compound can be separated into several distinct steps:
●● Dissolution of the solid using a single-solvent or a mixed-solvent system.
●● Decolorisation using charcoal: even if your compound is white, decolorisa-
tion will improve its appearance significantly.
●● Hot filtration (p. 134) to remove solvent-insoluble impurities and charcoal.
●● Cooling, to produce the crystals.
●● Collection of the crystals by suction filtration (p. 86).
●● Drying (p. 95).

Recrystallisation
Box 14.1 How to carry out a solvent selection for recrystallisation of an unknown compound
A suitable range of solvents for general use, in order of (b) The solid dissolves and thus the solvent may be
decreasing polarity, is: water, ethanol, propanone, ethyl useful for recrystallisation.
ethanoate, dichloromethane, toluene and light petroleum (c) Most of the compound dissolves but leaves a
(b.pt. 40–60 °C). small amount of insoluble residue. This means
that there are solvent-insoluble impurities pres-
1. Clean and dry seven Pyrex® test tubes to ensure ent in your compound and the solvent may be
that you will have no problems with contaminated suitable for use in recrystallisation.
solvents when carrying out the solubility tests. (d) The solid melts and floats on the meniscus at the
2. Add a small sample of the compound under test to top of the solvent giving the appearance of hav-
each tube, using just enough compound to cover ing dissolved. This is a common feature of low-
the bottom of the test tube. melting-point hydrocarbons such as naphthalene
and other non-polar aromatic compounds: shake
3. Add about 2.0 mL of a pure solvent to the first tube the test tube to see if oily globules are released
and observe the effect. You should look for the fol- from the meniscus. If this occurs, the solvent is
lowing features, which may help in the next recrys- unsuitable for recrystallisation.
tallisation stage:
5. Cool the test tube in an ice-water bath or under a
(a) The solid does not dissolve, but it is ‘wetted’ by cold stream of water, ensuring that no water gets into
the solvent. This implies that the solvent may the test tube. There are several possible results:
be suitable for use in recrystallisation.
(b) The compound dissolves easily. Therefore the (a) The compound recrystallises in high yield on
solvent is unsuitable for recrystallisation and there cooling: often there will appear to be more solid
is no need to continue tests with this solvent. than you started with because of the fine crystals
(c) Most of the compound dissolves but leaves a produced. This solvent is suitable.
small amount of insoluble residue. This means (b) The compound recrystallises in low yield on cool-
that there are solvent-insoluble impurities pres- ing: the solvent is unsuitable unless no better
ent in your compound, but the solvent itself is alternative can be found.
unsuitable for use in recrystallisation. (c) The compound remains in solution after cooling:
(d) Some colour is released into the solvent and the solvent is unsuitable or a supersaturated
the compound becomes lighter in colour. This solution has been formed. To check if the forma-
means that there could be a coloured impurity tion of a supersaturated solution has occurred,
present in your compound and it will be neces- gently scratch the inside of the test tube at the
sary to decolorise the product in the recrystalli- surface of the solution with a Pyrex® rod. The
sation experiment. scratches provide points of nucleation for crys-
tal growth and crystals should form rapidly if
4. Heat the mixture in the test tube by an appropriate the solution is supersaturated. Take care when
means, bearing in mind the flammability and toxic- scratching the test tube with the glass rod (see
ity of the solvent, to see if the solid dissolves in the p. 136). If a supersaturated solution is formed,
hot solvent. For non-flammable solvents, remember then the solvent is suitable for recrystallisation.
to hold the test tube with a holder (see p. 92) and to
‘wave’ the test tube over the heat source to prevent 6. Repeat the test using the other solvents and record
‘bumping’ (p. 87). Flammable solvents should be your results in tabular form to include your experi-
heated using a steam bath in a fume cupboard: do mental observations and conclusions.
not lower the test tube into the steam bath and leave
it there. Instead you must ‘wave’ the test tube in the 7. If you have found several suitable solvents, then
steam escaping from the bath to achieve controlled select one after considering factors such as flam-
heating. This is particularly important with low-boiling- mability, toxicity and boiling point, since you will
point flammable or toxic solvents such as propanone, need to use much larger volumes of solvent in the
dichloromethane and light petroleum (b.pt. 40–60 °C). recrystallisation process, with consequent compli-
You should look for the following results: cations in the equipment to be used (see Table 14.2).
(a) The solid does not dissolve in the hot solvent, 8. If you have not found a suitable solvent, then you
which is therefore unsuitable for recrystallisation will need to carry out a mixed-solvent recrystallisa-
and there is no need to continue tests with this tion and you must find out which combination of sol-
solvent. vents will be suitable (see Box 14.2).

Recrystallisation
Box 14.2 H
ow to carry out a mixed-solvent selection for recrystallisation of an unknown
compound
From your solvent selection tests (Box 14.1) you will have 6. Repeat the heating and solvent addition pro-
discovered the individual solvents in which your compound cess until the point is reached when the precipitate
is soluble (good solvents) and in which it is insoluble (poor just dissolves when the solution is heated. You now
solvents), when the solvents are cold. Proceed as follows: have a hot solution of your compound with the cor-
rect ratio of the two solvents.
1. Choose a miscible solvent pair, in which the solubil-
ity of your compound is appropriate. 7. If you have added too much poor solvent and the
precipitate will not redissolve on heating, add
2. Place a small amount (about 0.2 g) of the compound enough good solvent to dissolve the precipitate
in a clean dry Pyrex test tube and add a good sol-
®
and then continue to add poor solvent and heat as
vent (about 2.0 mL) in which it is soluble.
before.
3. Heat the test tube by an appropriate method, depend- 8. Cool the test tube in an ice–water bath, to ensure
ent on the solvent used, until the solid has dissolved.
that crystals are formed.
Remember that your original solvent selection may
have shown that solvent-insoluble impurities are 9. If an oil is formed on cooling, which may happen
present. if the melting point of the solid is below the boiling
points of the solvents used, you should use slightly
4. Add a few drops of the poor solvent from a Pasteur more of the good solvent (or less of the poor solvent)
pipette. A slight ‘cloudiness’ or precipitate should
or try a different combination of solvents.
form, since the solubilising power of the good sol-
vent has been decreased by the poor solvent and the 10. If crystals do not form on cooling, you may have
solution will have been cooled slightly. formed a supersaturated solution and you should
‘scratch’ with a Pyrex® glass rod, as described for
5. Reheat the solution until the cloudiness or precipi- single solvent selection (Box 14.1).
tate disappears and then add a few more drops of
the poor solvent to produce a precipitate.
It is important to remember that for a successful recrystallisation, you need

to use equipment of a size appropriate to the amount of solid and the volume of
solvent you are likely to use. You can estimate the volume of solvent to be used
by extrapolation of the data from your solvent selection tests. In general terms,
conical flasks, beakers and round-bottom flasks should never be more than
half-full of solution but, on the other hand, using small volumes of solutions
in large flasks will result in losses of the compound on the sides of the vessels.
Dissolution
To carry out a single-solvent recrystallisation (Box 14.3) you must get the
compound into solution and this is achieved by suspending it in the appropri-
ate cold solvent, found in the solvent selection process, and then heating the
mixture to dissolve the solid. The equipment used will depend on the boiling
point of the solvent, its flammability and toxicity. Some general systems are
shown in Table 14.2.
When heating solvents in conical flasks and beakers you should cover
the top of the flask with a clock-glass or watch-glass to prevent excessive
evaporation of the solvent, resulting in the formation of crystals on the sides
of the flask above the surface of the solution. Do not forget to take the appro-
priate ‘anti-bumping’ precautions (p. 87), because you may need to boil the
mixture for several minutes to achieve complete dissolution of the chemical(s).
You should use a glass rod, a wooden ‘boiling stick’ or anti-bumping chips

Recrystallisation
Box 14.3 How to carry out a single-solvent recrystallisation
Examples: Suppose that you have prepared a crude sam- the recrystallisation solution hot during the filtration
ple (about 4.0 g) of N-phenylethanamide (acetanilide) by putting it back onto the hot plate.
and you are to purify if by recrystallisation from water.
The melting point of pure N-phenylethanamide is 114 °C 8. When filtration is complete, remove the collection
(Haynes, 2015). flask from the heat, take out the glass rod and clamp
the flask in an ice–water bath, covering the top of the
1. Weigh the crude sample and retain a few crystals in flask with a watch-glass.
case seeding is required.
9. When the solution is cold (about 5 °C), collect the
2. Transfer the solid, using glazed paper or a solids solid by suction filtration (Box 10.2), using the filtrate,
funnel (p. 80), into a clean, dry conical flask (250 mL). not fresh water, to transfer the solid completely from
the collection flask. If crystals do not form, either add
3. Add cold water (about 100 mL) and a glass rod as a few crystals of the crude solid to ‘seed’ the super-
an anti-bumping device to the flask and then heat saturated solution or ‘scratch’ with a Pyrex® glass rod
the mixture on a hot plate until the compound has to induce recrystallisation.
dissolved completely. Then add more water (about
10mL) to ensure that precipitation of the solute does 10. Rinse the compound on the filter using a little
not occur during the following stages. (about 5 mL) ice-cold water and continue suction, to
make the crystals as dry as possible.
4. Remove the flask from the heat and allow it to cool
for 2 minutes. 11. Transfer the crystals to a clock-glass using a spatula
and spread them in a thin layer.
5. Add a small amount of decolorising charcoal (about
0.01 g) to the solution, place a watch-glass on top 12. Dry the pure compound in the oven at 70 °C for
of the conical flask and heat the mixture gently for about 30 minutes and test them with a spatula to
5 minutes. check their dryness: they should not stick together.
6. Prepare another clean, dry conical flask (250 mL) and 13. Allow the crystals to cool, covering them with
another clock-glass to prevent contamination.
add water (about 20 mL) and a glass rod and then
heat to boiling on the hot plate with a stemless 14. Transfer the purified crystals to a ‘tared’ watch-
funnel and fluted filter paper just above the neck glass on a balance to determine their weight and
(Fig. 14.1), so that the steam heats the funnel and then transfer them to a labelled sample tube using
filter paper. folded glazed paper (p. 80).
7. Filter the recrystallisation solution through the filter 15. Calculate the efficiency of the process:
paper using hand protection (rubber fingers or an
insulated glove) and keep the filter topped up with weight of pure compound
% recovery = * 100
solution to prevent cooling. At the same time, keep weight of crude compound
Table 14.2 Solvents for recrystallisation
Solvent b.pt. (°C) Glassware Heat source Containment
Water 100 Conical flasks, Burner, hot None

beakers plate
Ethanol 78 Conical flasks Water bath Fume cupboard
Propanone 56 Conical flasks Water bath Fume cupboard
Ethyl ethanoate 78 Conical flasks Water bath Fume cupboard
Dichloromethane 41 Reflux Water bath Fume cupboard
Toluene 110 Reflux Hot plate, Fume cupboard
mantle
Light petroleum 40–60 Reflux Water bath Fume cupboard

Recrystallisation
in conical flasks of volume up to 500 mL; magnetic ‘fleas’ or anti-bumping

granules in round-bottom flasks in reflux apparatus (p. 87); magnetic ‘fleas’
or stirring bars in large-volume glassware (7 500 mL).
Decolorisation
Coloured compounds – you should Small amounts of coloured impurities can be removed from your product
know something about the colour of by absorption on finely divided charcoal, which is then removed in the hot
the compound you are trying to purify. filtration process. In general, you should use charcoal in every recrystalli-
There is no point in trying to remove sation since the colour of white and coloured compounds is improved by
the colour from a coloured compound. ‘decolorisation’.
However, charcoal will improve signif- The amount of charcoal used should be about 2% by weight of the sample
icantly the appearance of a coloured to be recrystallised. Note the following important SAFETY POINTS:
compound.
●● Add the charcoal only when the solution has been formed. If you add char-
coal to the cold solution, you will not be able to see when all of the com-
S o l ve n t - i n s o l u b l e i m p u ri t i e s –
pound has dissolved.
remember that if you have found
solvent-insoluble impurities, not all
●● Do not add charcoal to a boiling or a very hot solution otherwise the solution
your ‘compound’ will dissolve in the will boil extremely vigorously, usually boiling out of the flask or reflux
hot solvent. You can confirm the pres- apparatus.
ence of these by adding an excess of
●● To add charcoal to a hot solution, remove the flask from the heat source and
hot ‘selected’ solvent to a small sample
then let it cool a little (not enough to cause precipitation). Add the charcoal
of the crude compound. If it does not
either from a spatula (into a conical flask or beaker), a paper funnel (p. 80)
dissolve completely, solvent-insoluble
impurities are present.
or powder funnel (into a round-bottom flask with a ground-glass joint in
reflux apparatus), and then reheat the solution.
Hot filtration
This is a modification of gravity filtration (p. 84), designed to remove
solvent-insoluble impurities, charcoal, anti-bumping granules or magnetic
‘fleas’ from the hot solution before cooling the solution to form the crystals
of purified product.
Key Point Hot filtration is used to prevent cooling of the

solution during the filtration process, which would result in the
formation of crystals in the filter paper.
hand protection
essential
For a successful hot filtration the solution must pass through the filter
paper and filter funnel into the collection flask as quickly as possible so
that cooling and crystallisation do not occur. The following points should
be noted:
●● Always use a fluted filter paper (p. 84).
stemless funnel
●● Always use a ‘stemless’ glass filter funnel, because cooling and thus crystal-
lisation may occur in the funnel stem causing a blockage.
hotplate ●● Always heat the filter funnel and filter paper, either by pre-heating them in an
filtrate refluxing (or water bath)
gently oven or by using boiling solvent in the collecting flask (Fig. 14.1).
●● If filtration is rapid, keep the filter cone ‘topped up’ with the hot solution
being filtered, since this keeps the filter cone and filter funnel hot. If
you allow the filter cone to empty of liquid, cooling and crystallisation
Fig. 14.1 Filtration of a hot solution. may occur.

Recrystallisation
●● When attempting a hot filtration with a mixed-solvent system, always ensure

that the ‘ideal’ solvent ratio has not been reached, i.e. there is not enough of
the poor solvent to cause immediate precipitation as a result of slight cooling
during the hot filtration. The solvent ratio can be adjusted to the ‘ideal’ ratio
for maximum recovery, after filtration and before cooling (Box 14.4).
Box 14.4 How to carry out a mixed-solvent recrystallisation
Examples: Suppose that you have prepared a crude sam- (Box 14.1) so that the ethanol vapour heats the funnel
ple (e.g. ∼ 4.0 g) of N-phenylbenzamide (benzani-lide). and filter paper.
Your solvent selection tests have shown that it is insolu-
ble in water and fairly soluble in cold ethanol. The melt- 8. Filter the recrystallisation solution through the filter
ing point of pure N-phenylbenzamide is 158 °C (Haynes, paper using hand protection (rubber fingers or an
2015). Proceed as follows: insulated glove) and keep the filter topped up with
solution to prevent cooling. At the same time, keep
1. Weigh the crude sample and retain a few crystals in the recrystallisation solution hot during the filtration
case seeding is required. by putting it back onto the hot plate.
2. Transfer the solid, using glazed paper or a solids 9. When filtration is complete, remove the filter funnel
funnel (p. 80), into a clean, dry conical flask (250 mL). and restart the water addition and reheating process
until the ideal solvent ratio is reached once again.
3. Add cold ethanol (about 25 mL) and a glass rod as
an anti-bumping device to the flask and then heat the 10. Remove the collecting flask from the heat, take out
mixture on a water bath until the compound has dis- the glass rod and clamp the flask in an ice–water bath,
solved completely. covering the top of the flask with a watch-glass.
4. Add cold water (about 5 mL) to the hot ethanol solu- 11. When the solution is cold (about 5 °C), collect the
tion and a slight precipitate should form, which then solid by suction filtration (Box 10.2), using the filtrate,
redissolves – this is often seen as a ‘flash’ of white in to transfer the solid completely from the collection
the solution. Reheat the solution and add more water flask. If crystals do not form, either add a few crystals
(5 mL). The precipitate will remain for a little longer of the crude solid to ‘seed’ the supersaturated solu-
and take more time to disappear on reheating. tion or ‘scratch’ with a Pyrex® glass rod to induce
recrystallisation.
5. Repeat the water addition and reheating pro-
cess, but note that you will need to reduce the vol- 12. Rinse the compound on the filter using a little
ume of water added at each consecutive addition, (about 5mL) ice-cold water and continue suction, to
since the precipitate will take longer to redissolve as make crystals as dry as possible.
the solvating power of the ethanol is reduced by the
water. Eventually, you will reach the situation where 13. Transfer the crystals to a clock-glass using a spatula
the precipitate just redissolves at the boiling point of and spread them out in a thin layer.
the solvent mixture. You now have the ideal solvent 14. Dry the pure compound in the oven at 70 °C for
mixture for recrystallisation of N-phenylbenzamide about 30 minutes and test them with a spatula to
and any slight cooling of the solution will result in its check their dryness: they should not stick together.
precipitation.
15. Allow the crystals to cool, covering them with
6. Remove the solution from the heat, add etha- another clock-glass to prevent contamination.
nol (5 mL), to ensure that the N-phenylbenzamide
remains in solution if it cools a little, add charcoal 16. Transfer the purified crystals to a ‘tared’ watch-
(about 0.01 g), place a watch-glass on top of the flask glass on a balance to determine their weight and
and reheat the solution on the water bath for about 5 then transfer them to a labelled sample tube using
minutes. folded glazed paper (p. 80).
7. Prepare another clean, dry conical flask (250 mL) and 17. Calculate the efficiency of the process:
add ethanol (about 10 mL) and a glass rod and then
weight of pure compound
heat to boiling on the water bath with a stemless % recovery = * 100
funnel and fluted filter paper just above the neck weight of crude compound

Recrystallisation
Cooling
Rapid cooling in an ice–water bath (‘crash-crystallisation’) usually produces
small crystals occluded with mother-liquor, whereas slow cooling by allow-
ing the collection flask to stand on the laboratory bench often produces large
well-defined crystals. Remember to:
●● cover the top of the collection flask with a watch- or clock-glass to prevent
solvent evaporation and entry of dust into the flask: do not use rubber bungs
or corks since they may be pulled into the flask as it cools (p. 76);
●● clamp the flask in place, if you use an ice–water bath, otherwise it may fall
over as the ice melts and the volume of water increases;
●● make sure that the solution is cold, even after slow cooling, so that maximum
precipitation of the solid occurs.
Safety Note Take great care when
‘scratching’ a supersaturated solution If no crystals appear on cooling, you will have formed a supersaturated solu-
with a glass rod. Hold the flask or test tion and, to induce precipitation of the solute, you must provide sites for nucle-
tube at the neck, not at the bottom, ation and crystal growth. This can be achieved by either seeding the solution by
and make short scratching movements adding a few crystals (‘dust’) of the crude compound or scratching the inside of
on the side of the flask just above the flask at the surface of the liquid, using a Pyrex® glass rod (Fig. 14.2).
and below the surface of the liquid Collection of the crystals
(Fig. 14.2). If you accidentally break the
flask or test tube you will cut your hand
You should collect the purified crystals by suction filtration by the procedure
if you hold the vessel at the bottom.
described in Box 10.2 (p. 86). Remember to transfer the crystals from the col-
lecting flask to the filter using a little of the filtrate: do not use fresh solvent.
(a) (b)
(c)
Fig. 14.2 ‘Scratching’: (a) right; (b) right; (c) wrong.

Recrystallisation
The filtrate is a saturated solution of the compound being recrystallised and

cannot dissolve any more solute, but fresh solvent will dissolve some of your
product, resulting in an inefficient recrystallisation process.
Drying
The purified compound should be dried by the appropriate method as described
on p. 95. If you do not know the melting point of your compound, you should
always carry out a test by placing a small amount on a watch-glass in the oven,
before committing the bulk of your chemical.
Detailed procedures for single-solvent and mixed-solvent recrystallisa-
tions are shown in Box 14.3 and Box 14.4, respectively and the modifications
necessary for the use of other solvent systems can be worked out from the
information in Table 14.2.
Problems in recrystallisation
Crystallisation in the filter paper:
There are three common problems encountered during recrystallisation:
prevention – keep everything hot and
use a good fluted filter paper; 1. The compound crystallises in the filter funnel during hot filtration. This is
because the solubility of the solute decreases rapidly with temperature and
cure – wash through with hot sol-
the slight cooling during hot filtration causes precipitation of the solid, even
vent, evaporate most of the excess and
though you are heating the funnel. The answer is to use more than the min-
repeat the hot filtration.
imum amount of solvent and then evaporate off the excess before cooling.
2. The compound does not recrystallise. There are two reasons: you have
Evaporation of excess solvent –
used too much solvent and you must evaporate off some solvent before
remove by rotary evaporation (p. 161)
cooling, or you have formed a supersaturated solution and you must ‘seed’
or N2 blowdown (p. 163). This is the one
or ‘scratch’ the solution (see p. 136).
occasion when water is the less than 3. The compound precipitates as an oil. This is because compounds with low
ideal solvent, since its evaporation is melting points often come out of a concentrated solution above their melt-
time consuming. ing point. In such cases more solvent should be added and the compound
redissolved and cooled so that precipitation is retarded to the temperature
at which the crystalline solid comes out of solution. Often ‘scratching’ the
hot solution as it cools can prevent ‘oiling out’.
Text references
Amarego, W.L.F. and Chai, C. (2012) Purification of Lab- Isac-Garcia, J., Dobado, J.A., Calvo-Flores, F.G.
oratory Chemicals, 7th edn. Butterworth-Heinemann, and Martinez-Garcia, H. (2015) Experimental
London. Organic Chemistry. Laboratory Manual. Elsevier,
Amsterdam.
Buckingham, J. and Macdonald, F. (eds) (1995) Dictionary
of Organic Compounds, 6th edn. CRC Press, Boca Raton. Leonard, J., Lygo, B. and Procter, G. (2013) Advanced
Practical Organic Chemistry, 3rd edn. CRC Press, Boca
Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch-
Raton.
ell, A.R. (1991) Vogel’s Textbook of Practical Organic
Chemistry, 5th edn. Longman, Harlow, Essex. Lehman, J.W. (2009) Student Lab Companion: Laboratory
Techniques for Organic Chemistry, 2nd edn. Pearson,
Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper-
Harlow.
imental Organic Chemistry, 2nd edn. Blackwell Science
Ltd, Oxford. Loewenthal, H.J.E. and Zass, E. (1992) A Guide for the
Perplexed Organic Experimentalist, 2nd edn. John

Recrystallisation

Keese, R., Brandle, M.P. and Toube, T.P. (2008) Practical Zubrick, J.W. (2015) The Organic Chem Lab Survival
Organic Synthesis: A Student’s Guide. John Wiley & Manual. A student’s guide to techniques, 10th edn. John
Sons Ltd, Chichester. Wiley & Sons Ltd, Chichester.
Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical
Organic Chemistry. Chapman & Hall, London.
Williamson, K.L. and Masters, K.M. (2010) Macroscale
and Microscale Organic Experiments, 6th edn. Brooks
Cole, Boston.
Study exercise
14.1 Prepare a list of all the laboratory equipment small amount of grit, which should have a m.pt.
you would require to recrystallise, from ethanol, ∼ 140 °C.
an off-white solid ( ≈4 g) contaminated with a

15 Solvent extraction
This technique separates the components of chemical mixtures by using the

Extraction – making a cup of coffee
dissimilar solubility properties of the components of the mixture in different
involves extraction of the flavour chem-
solvents. Extraction is used mainly to purify a reaction product partially before
icals and caffeine from the insoluble
final purification by recrystallisation (p. 128) or distillation (p. 145). The two
vegetable matter using hot water and
common types of extraction process used in the laboratory are:
is an example of liquid–solid extraction.
1. Liquid – liquid extraction: this uses two immiscible solvents; the desired
compound in solution or suspension in one solvent is extracted into the
other solvent. For example, covalent organic compounds are extracted
from aqueous solution into dichloromethane, leaving the ionic by-products
or reagents in the aqueous phase.
2. Solid–liquid extraction: this involves the use of a solvent to remove sol-
vent-soluble components of a solid mixture.
Liquid–liquid extraction
Several experimental processes in practical chemistry are based on liquid–
liquid extraction:
●● ‘Extraction’: where a solid or liquid suspended or dissolved in one solvent
is extracted into another. This technique can be used to separate covalent
molecules from ionic compounds in an aqueous solution or suspension.
●● ‘Washing’: where ionic species are removed from a non-polar solvent by
extraction into water.
●● ‘Acid–base extraction’: where covalent molecules are converted into their
salts and thus removed from a non-polar solvent into water, while neutral
covalent species will remain in the non-polar solvent, as shown in Table 15.1.
All of these processes involve mixing the two immiscible solvents, one
of which contains the mixture, in a separatory funnel (p. 140) and shaking the
funnel to promote the extraction process. The immiscible layers are allowed to
reform and are then separated.
Table 15.1 Examples of acid–base extraction chemistry
ArCOOH + RH NaOH
¡ ArCOO- Na + + RH
CH2Cl2
Acid Neutral Salt Neutral

insoluble in H2O insoluble in H2O soluble in H2O insoluble in H2O
soluble in CH2Cl2 soluble in CH2Cl2 soluble in CH2Cl2
ArNH2 + RH ¡
HCl
ArNH3+ Cl - + RH
CH2Cl2
Amine Neutral Salt Neutral

ArCOOH + ArOH Na2CO3
¡ ArCOO- Na + + ArOH
CH2Cl2
Acid Weak acid Salt Weak acid


Solvent extraction
For liquid–liquid extraction, water is usually the polar solvent.

Safety Note Because of its flam- Since most extractions involve getting the required compound into the
mability and tendency to form explo- organic solvent (or removing unwanted ionic chemicals from it), it should
sive peroxides, the use of diethyl ether have good solvent power for the desired compound and a low boiling point
for extractions in many undergraduate for ease of removal and recovery of the compound. The common organic
laboratory has effectively been discon- solvents used in liquid–liquid extraction are diethyl ether (ethoxyethane)
tinued. Use dichloromethane or ethyl b.pt. 34 °C, dichloromethane (DCM) b.pt. 41 °C and ethyl acetate (ethyl
ethanoate instead. ethanoate) b.pt. 77 °C. Dichloromethane is denser than water and forms
the lower layer, whereas diethyl ether and ethyl acetate float on water and
are the upper layer.
Partition coefficients
Safety Note Dichloromethane is The theory of liquid–liquid extraction is based on the equilibrium between the
effectively non-flammable and is often concentrations of dissolved component in the two immiscible liquids, when
the solvent of choice, but it is a liver they are in contact. The equilibrium constant for this process is called the par-
toxin and an irritant, and must be han- tition coefficient or distribution coefficient and is given by:
dled in the fume cupboard if volumes
greater than 50 mL are to be used. concentration of solute in liquid 1
K = [15.1]
concentration of solute in liquid 2
You only need to calculate such quantities if:
●● you are carrying out specific experiments to determine partition coefficients,
when you will be given specific instructions or references to the appropriate
literature;
●● the solute has appreciable solubility in both solvents.
The reason calculation is not necessary is that, in the overwhelming major-
ity of extractions you will carry out, the conditions used are designed to ensure
that the components will be almost totally soluble in one of the liquids and
Extraction calculations – it is necessary almost insoluble in the other, since complete separation is required. The num-
to calculate volumes of solvents and ber of extractions needed to extract a water-soluble solute into an immiscible
number of extractions when attempting organic phase can be calculated from the following relationship:
to maximise the economics of an indus- n
trial-scale extraction process. Kn
wn = w0 a b [15.2]
Kn + s
where K = partition coefficient of the solute, n = volume (mL) of aque-
ous solution of the solute, s = volume (mL) of immiscible organic solvent,
Separatory funnels – the cone-shaped w0 = weight of solute initially in the aqueous layer, wn = weight of solute
funnels are specifically designed for remaining in the aqueous layer after n extractions, and n = number of extrac-
extractions. Only use parallel-sided fun- tions. Evaluation of this expression shows that, for a fixed volume of solvent,
nels when no alternative is available. it is more efficient to carry out many small extractions than one big one.
Separatory funnels
These come in a range of sizes from 5 mL to 5000 mL and there are two
general types: parallel sided and cone shaped (Fig. 15.1). Cone-shaped
separatory funnels are made of thin glass and should be supported in a
Safety Note Extractions involving
ring (p. 141). Small-volume cone-shaped funnels ( 6 100 mL capacity) and
large volumes of solvents ( 7 100 mL)
parallel-sided separatory funnels should be clamped at the ground-glass
should always be carried out in cone-
joint at the neck (p. 82).
shaped separatory funnels supported
Separatory funnels will have glass or Teflon® taps with a rubber ring and
by a ring, because there is then no dan-
clip or screw cap on the end to prevent the tap slipping from the barrel, or a
ger of the funnel slipping from a clamp
Rotaflo® tap. You must ensure that the tap assembly is in good condition by
at its neck.
making the following checks before starting work:

Solvent extraction
●● For glass taps: disassemble the tap by first removing the clip and ring or cap
from the tap (note the order of the component parts for reassembling). Dry
the tap and barrel with tissue, add a light smear of grease to the tap (making
sure you do not clog the hole in the tap) and reassemble the tap and fittings,
turning the tap to ensure free movement. Support the separatory funnel in
position and add some of the organic solvent to be used (2–3 mL) to the
funnel, with the tap closed, to check that the tap does not leak. If the tap
leaks, disassemble and regrease.
●● For Teflon® taps: disassemble the tap, wipe the tap and barrel with clean
tissue, reassemble without grease and check for free movement of the tap and
for leakage as described above. When you have finished using the funnel,
glass or Teflon® tap loosen the clip/cap on the tap since Teflon® will flow under pressure and
the tap may ‘seize’ in the barrel.
●● For a Rotaflo® tap: unscrew the tap from the funnel and ensure that the
plastic locking thread is in place (Fig. 15.2). If it is not present, consult your
instructor and obtain a replacement. Dry the barrel of the tap and the tap with
a tissue and reassemble. Do not grease the Rotaflo® tap.
The general procedure for using a separatory funnel for extraction is
described in Box 15.1 and there are five additional practical tips to aid your
success:
Rotaflo® tap
1. Label all flasks to avoid confusion.
Fig. 15.1 Separatory funnels. 2. Never throw away any of the separated liquids until you are absolutely
sure of their identity.
3. Always transfer solvents into the separatory funnel using a stemmed
Drying separatory funnels in an oven – filter funnel so that solids and liquids will not stick to the inside of
always disassemble the tap and do not the joint and prevent a good seal when you insert the stopper and then
place the tap and its plastic components invert the funnel.
in the oven. Dry them with tissue.
4. Always place a ‘safety’ beaker under the separatory funnel to collect liquid
just in case the tap leaks (Fig. 15.3(a)).
funnel inverted stopper
separated liquid layers

covered metal ring
(e.g. PVC)
plastic locking ring

stand stopcock (closed!)
glass thread
beaker
stopper
(ready for use)
Rotaflo® tap (a) (b)
Fig. 15.2 A Rotaflo® tap. Fig. 15.3 A separatory funnel (a) ready to use and (b) in use.

Solvent extraction
Box 15.1 How to separate a carboxylic acid and a hydrocarbon using solvent extraction
This is an example of an acid–base extraction. The solid 10. Take out the stopper, place it upside down in the top of
mixture (e.g. 4.0 g for the solvent volumes used below) the separatory funnel and allow the solvent layers to
of benzoic acid and naphthalene is soluble in dichlo- separate. The upper layer is the aqueous layer (10 mL
romethane but benzoic acid will dissolve in dilute aque- compared with 50 mL of dichloromethane). Sometimes
ous sodium hydroxide (2 M) by forming the sodium salt a few globules of dichloromethane will ‘cling’ to the sur-
(sodium benzoate). Naphthalene is insoluble in water. face of the water layer: these can be released by gently
swirling the contents of the separatory funnel.
1. Dissolve the mixture in a clean, dry beaker in dichlo-
romethane (50 mL). 11. When the liquids have stopped swirling, open the
tap gently and slowly run the dichloromethane lower
2. Clean and dry the tap of a separatory funnel (250 mL) layer into a clean conical flask and label it ‘dichlo-
and set up as shown in Fig. 15.3(a).
romethane layer’. Avoid fast emptying of the funnel
3. Make sure that the tap is closed and then add the because a vortex may be formed which will cause the
solution containing the mixture, using a stemmed upper layer to run out with the lower layer.
funnel to prevent contamination of the joint, and rinse
12. Run the remaining aqueous layer into a clean, dry
out the beaker with dichloromethane (∼ 5 mL).
conical flask and label it ‘sodium hydroxide layer’.
4. Add sodium hydroxide solution (10 mL) to the separa- 13. Return the dichloromethane layer to the separatory
tory funnel, place the stopper in the separatory funnel
funnel and extract it with another portion (10 mL) of
and gently invert it and hold it as shown in Fig. 15.4.
sodium hydroxide. Repeat the extraction process for a
Do not shake the separatory funnel since you do not
total of 40 mL (i.e. 4 * 10 mL) of the alkali, collecting
know how much heat will be produced in the reac-
all the sodium hydroxide extracts in the same flask.
tion, which will pressurise the separatory funnel.
14. Finally, extract the dichloromethane with water (20
5. Open the tap, to release any pressure caused by the mL), to remove any traces of sodium hydroxide, and
heat of reaction.
add these ‘washings’ to the sodium hydroxide layer’
6. Close the tap, shake the mixture once and open the flask.
tap to release any pressure.
15. You now have a solution of naphthalene in
7. Close the tap, shake the mixture twice and open the dichloro-methane and a solution of sodium benzoate
tap to release any pressure. in sodium hydroxide ready for further processing.
8. Repeat until no more vapour is expelled via the tap. 16. If an emulsion forms, the layers do not have a well
defined boundary: add a few drops of methanol to the
9. Close the tap, and replace the separatory funnel in upper layer down the inside wall of the funnel. This
the ring or clamp. often ‘breaks’ the emulsion.
tap
5. Always take the stopper from the separatory funnel before you attempt to
allow liquid to run from the funnel. If you do not remove the stopper from
the top of the funnel, a vacuum is formed in the funnel after a little of the
liquid has run out. Air will be sucked into the funnel through the outlet
stem causing bubbles, which will remix your separated layers. If your
funnel is equipped with a Quickfit® stopper, it is good practice to take
Fig. 15.4 Holding a separatory funnel. the stopper out of the top and put it back upside down (Fig. 15.3(b)). This
ensures that no vacuum is formed and that organic vapours do not escape
Batch liquid–liquid extraction – the pro- easily from the flask.
cess described is inefficient if the material
being extracted has appreciable solubility
in both of the solvents used. In these sit- Solid–liquid extraction
uations a continuous extraction system is In this process the components of a solid mixture are extracted into a solvent.
necessary and you should consult the spe- The ‘batch process’, analogous to liquid – liquid extraction, involves grinding
cialist textbooks, e.g. Furniss et al. (1991). the solid to a fine powder, mixing it with the appropriate solvent, and filtering

Solvent extraction
Box 15.2 How to set up a Soxhlet extraction system
1. Select apparatus of the appropriate size for the 7. Half-fill the extraction thimble with the solid to be
amount of solid to be extracted. Specifically, the Sox- extracted and plug the top of the thimble with white
hlet thimble should fit below the siphon outlet and cotton wool to prevent any solid being carried over
the volume of the solvent reservoir should be such into the solvent.
that it is never more than half-full when all the solvent
has siphoned from the extractor. 8. Place the thimble in the extractor.
2. Assemble the apparatus as shown in Fig. 15.5, clamp- 9. Attach a water supply to the reflux condenser, lightly
ing at the joints at the flask and the top of the Soxhlet grease the male joint and attach the condenser to
extractor. The best heat source to use for continuous the top of the extractor. Turn on the water, ensuring a
operation over a long period is a mantle. steady flow.
3. Disassemble the apparatus, leaving the clamps in 10. Switch on the heater and turn up the power so that
position. the solvent refluxes and drips into the extractor.
4. Fill the flask to about one-third of its volume with 11. Confirm that everything is running smoothly by
solvent, add some anti-bumping granules (or a mag- watching at least two siphoning cycles of the extrac-
netic ‘flea’, if a stirrer-mantle is being used) and clamp tion and check the apparatus frequently.
it into position in the mantle.
12. When the extraction is complete, allow the apparatus
5. Lightly grease the ‘male’ joint of the Soxhlet extrac- to cool and dismantle it. Place the extraction thimble
tor and attach it to the reservoir flask. Clamp the top in the fume cupboard to allow the solvent to evapo-
of the extractor. rate and then dispose of it appropriately. Gravity filter
or decant the solvent in the reservoir flask to remove
6. Add solvent to the extractor until it siphons. This the anti-bumping granules of magnetic ‘flea’ and
ensures that the reservoir will never be dry. Check remove the solvent by distillation (p. 145) or rotary
that the reservoir is now no more than half-full; if it evaporation (p. 163).
is, replace with a larger flask.
off the solid by gravity (p. 83) or under vacuum (p. 84) and then evaporating
condenser
the solvent (p. 161) from the extract solution. However, a more elegant ‘contin-
uous extraction process’, called Soxhlet extraction, is available when the most
appropriate solvent is known.
water in The apparatus for Soxhlet extraction is shown in Fig. 15.5 and comprises a
clamp
flask containing the solvent, a Soxhlet extractor and a reflux condenser (p. 155).
paper The solid to be extracted is placed in a porous thimble, made from hardened
thimble
solid to be Soxhlet
filter paper, and the solvent is heated so that its vapour flows past the thimble,
extracted apparatus condenses and fills the extractor with hot solvent to extract the solid. When the
extractor is full, the solvent (together with the extracted material) siphons back
into the solvent flask and the process is repeated automatically. The advantage
of this procedure is that fresh solvent continually extracts the solid, which is
clamp concentrated in the flask. The disadvantage is that the compound extracted is
kept at the boiling point of the solvent for a prolonged period. Soxhlet extrac-
tors come in sizes of 10 mL to 5000 mL, based on the volume of solvent con-
boiling tained in the extractor. The procedure for using a Soxhlet extraction system is
heat source stones described in Box 15.2.
(mantle, water
bath, etc.)
Fig. 15.5 A Soxhlet extraction system.

Solvent extraction
Text reference
Amarego, W.L.F. and Chai, C. (2012) Purification of Lab- Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch-
oratory Chemicals, 7th edn. Butterworth-Heineman, ell, A.R. (1991) Vogel’s Textbook of Practical Organic
London. Chemistry, 5th edn. Longman, Harlow, Essex.

Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper- Keese, R., Brandle, M.P. and Toube, T.P. (2008) Practical
imental Organic Chemistry, 2nd edn. Blackwell Science Organic Synthesis: A Student’s Guide. John Wiley &
Ltd, Oxford. Sons Ltd, Chichester.
Isac-Garcia, J., Dobado, J.A., Calvo-Flores, F.G. and Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical
Martinez-Garcia, H. (2015) Experimental Organic Organic Chemistry. Chapman & Hall, London.
Chemistry. Laboratory Manual. Elsevier, Amsterdam. Smart, L. (ed.) (2002) Separation, Purification and Identi-
Lehman, J.W. (2009) Student Lab Companion: Laboratory fication – The Molecular World. Open University and
Techniques for Organic Chemistry, 2nd edn. Pearson, Royal Society of Chemistry, Cambridge.
Harlow. Williamson, K.L. and Masters, K.M. (2010) Macroscale
Leonard, J., Lygo, B. and Procter, B. (2013) Advanced Prac- and Microscale Organic Experiments, 6th edn. Brooks
tical Organic Chemistry, 3rd edn. CRC Press, Boca Raton. Cole, Boston.
Loewenthal, H.J.E. and Zass, F (1992) A Guide for the Per- Zubrick, J.W. (2015) The Organic Chem Lab Survival
plexed Organic Experimentalist, 2nd edn. John Wiley & Manual. A student’s guide to techniques, 10th edn. John
Study exercise
15.1 Find at least four errors in the diagram

below, which illustrates a student’s attempt at
liquid–liquid extraction.

16 Distillation
Distillation is used to separate the components of a liquid mixture by vapour-

Working with azeotropes – not all liq-
ising the liquids, condensing the vapours and collecting the liquid condensate.
uid mixtures can be separated by dis-
Separation is the result of the differing boiling points of the individual constit-
tillation. In some cases an azeotrope, a
uents of the mixture and the efficiency of the distillation column. You may be
mixture of the liquids of definite com-
required to purify a liquid by distillation, which involves the removal of small
position, which boils at a constant tem-
quantities of impurities, or to separate a mixture of liquids by fractional distil-
perature, is formed. For example, an
lation, each of which can then be purified by distillation.
azeotrope containing 95.5% ethanol and
You will meet several types of distillation process, each applicable to dif-
4.5% water boils at 78.15 °C, which is
ferent situations depending on the chemicals to be purified or separated.
below the boiling point of pure ethanol
(78.3 °C). Therefore 100% ethanol can- ●● Simple distillation: used for separating liquids, boiling below 200 °C at
not be obtained by distillation of etha- atmospheric pressure, from other compounds. For effective separation there
nol–water mixtures, even though their should be a difference in the boiling points of the components of at least
boiling points are about 22 °C apart. 80 °C.
●● Fractional distillation: used for separating components of liquid mixtures,
which have boiling points differing by more than 25 °C, at temperatures
below 200 °C.
●● Vacuum or reduced-pressure distillation: used for separating liquids boiling
above 200 °C, when decomposition may occur at the high temperature. The
effect of distilling at reduced pressure is to lower the boiling point of a liq-
uid. This technique can be applied to both simple distillation and fractional
Using steam distillation – in practice, distillation.
the decision to use this approach to sep-
arate the components of a mixture is
●● Steam distillation: used for separating mixtures of chemicals such as oils,
based on previous experience. Consult
resins, hydrocarbons, etc., which are essentially insoluble in water and may
your instructor for advice.
decompose at their boiling points. Heating the compounds with steam makes
them distil below 100 °C.
Distillation equipment
Setting up distillation apparatus – do Apparatus used for the various types of distillation has several general features:
not allow the support stands to move
during the distillation, since this will
●● Distillation flask: usually round bottom or pear shaped with one or two necks
allow the ground-glass joints connect-
(to allow a vacuum bleed to be fitted).
ing the still-head and condenser to sep- ●● Still-head: to hold the thermometer and to channel the vapour into the con-
arate. The hot vapours will then escape denser. For fractional distillation, the fractionating column is fitted between
into the laboratory instead of going the distillation flask and the still-head.
down the condenser.
●● Condenser: usually with circulating cold water.
●● Take-off adapter: to allow the distillate to run into the collecting vessel. For
Securing distillation components – vacuum distillation, the adapter will have a vacuum inlet tube and could
do not use plastic joint clips on the have three ‘arms’ to allow three fractions to be collected without breaking
‘hot end’ of a distillation apparatus the vacuum.
since they may melt and the joints may
separate.
●● Receiving (collection) vessel: this can be a test tube, a measuring cylinder, a
conical flask, or a round-bottom flask with a ground-glass joint.

Distillation
No matter what type of distillation you are attempting, it is essential that

you assemble the apparatus correctly, since you are dealing with hot, often
flammable, liquids and vapours. A typical simple distillation apparatus is
shown in Fig. 16.1 and the method of assembly is described in Box 16.1.
screwcap adapter
with thermometer
Key Point You must ensure that all the ground-glass joints are
seated properly and the apparatus is clamped firmly so that no
joint clips movement of the joints will allow vapours to escape.
Simple distillation
water The procedure for doing a simple distillation is described in Box 16.2 and you
out
should note the following points:
water
in ●● Do not distil to dryness, i.e. no liquid remaining in the distillation flask, since
this causes overheating and charring (decomposition) of the residue. Always
leave 1 or 2 mL of liquid in the distillation flask.
●● Initially, some liquid may distil while the temperature rises rapidly. This is
termed ‘forerun’ which is often a small amount of solvent from an extraction
Fig. 16.1 Apparatus for simple distillation.
process (p. 139), for example. Collect and then, when the temperature stabi-
lises, collect the desired compound in a clean receiving vessel.
●● Towards the end of the distillation, the temperature may begin to rise again:
Heating a distillation flask – unlike this will be the higher boiling impurity. Stop heating, quickly remove the
when using a microburner, it is difficult receiving flask containing your pure compound and collect these last few
to stop the heating process instantly drops, termed ‘tailings’, in a fresh receiving vessel.
when using an oil bath or mantle on a
‘labjack’. Therefore have a clean receiver Fractional distillation
flask ready to collect the ‘tallings’ as
soon as the temperature begins to rise. Simple distillation is not very effective in separating liquids unless their boil-
ing points are at least 80 °C apart and a better separation can be achieved if a
fractionating column is used. There are many types available (Fig. 16.2) and
the device brings the ascending vapours into contact with the condensing (in
the fractionating column) liquid and amounts to a succession of many simple
Collecting fractions – as the tempera- distillations in which the descending liquid strips the high-boiling component
ture begins to rise ‘between fractions’ from the ascending vapour. Overall, the lower boiling component distils first
you will have 2 or 3 seconds to change and cleanly. The column packing, usually glass beads, helices or ‘fingers’,
receiving vessels before the liquid runs gives a large surface area for contact of the ascending vapours and descending
from the top to the bottom of the con-
denser. Have three or four pre-weighed
receivers ready to hand.
Distillation in fume cupboards – it may

be necessary to insulate the fractionat-
ing column from the ‘wind’ by wrapping
it in aluminium foil, since the draught
prevents equilibration in the column.
Don’t forget to leave a ‘window’ in the
foil so that you can see the vapour con- open column Vigreux column Claisen flask Claisen–Vigreux flask
densing near the top of the column.
Fig. 16.2 Common fractionating columns.

Distillation
Box 16.1 How to assemble the apparatus for a simple distillation
In general laboratory work, you will usually be distilling receiving vessel in position, use a separate support
small volumes of liquids (∼ 25 mL). Standard glass joint- stand.
ware apparatus size 14/23 (p. 97) is appropriate. If you are
to carry out larger scale distillations, you must consider 6. Disassemble the apparatus in the reverse order to
the weight of the components, specifically the receiver assembly by opening the clamps – do not move the
flask, which should be supported on a ‘labjack’, since a clamps and stands.
joint clip may not be strong enough to hold a large flask 7. Add the liquid to be distilled to the distillation flask
(250 mL or larger). using a stemmed funnel, together with anti-bumping
1. Clamp the distilling flask firmly to a support stand granules or boiling stick or a magnetic ‘flea’. The flask
ensuring that it is at the correct height to allow the should not be more than 50% full. Reclamp the flask
heat source (microburner, oil bath or mantle on a ‘lab- in position.
jack’) to be removed if necessary. 8. Lightly grease the ‘male’ joint on the still-head and
2. Insert the still-head in the top of the flask. replace it in the flask.
3. Carefully line up the condenser on a clamp (attached 9. Insert a thermometer into a screwcap adapter (p. 100)
to the same support stand as the distilling flask and adjust so that the bulb is just below the outflow
(Fig. 16.1), if possible) and adjust the height and angle from the still-head. Lightly grease the joint on the
of the clamp so that the condenser slides onto the screwcap adapter and put it into the still-head.
still-head joint. Remember to have the non-moving 10. Fit the rubber tubing onto the condenser (p. 156),
jaw of the clamp at the bottom (p. 83) otherwise you lightly grease the joint on the still-head and refit the
will lift the condenser when you tighten the clamp condenser, clamping it firmly into place.
and break the joint on the still-head or condenser or
both. Carefully tighten the clamp and ensure that the 11. Connect the lower tube to the water tap so that the
joints are ‘seated’. water will flow up the condenser, and upper tube should
be routed into the sink. Make sure that there are no kinks
4. Attach the take-off adapter to the bottom of the con- in the rubber tubes and, for safety, feed them over the
denser with a joint clip. If you are using a Quickfit® protruding arms at the back of the clamps. Turn on the
flask as a receiving flask, you must use a v
acuum-type water tap gently and check that the water flows freely.
take-off adapter (p. 99) otherwise you will be distill-
ing in a closed system, which may explode when you 12. Lightly grease the bottom joint of the condenser and
begin to heat the distilling flask. attach the take-off adapter using a joint clip.
5. Attach the Quickfit® receiver flask to the take-off 13. Attach the Quickfit® receiving flask to the take-off
adapter with a joint clip or place the receiver flask adapter with a joint clip (after lightly greasing) or
underneath the take-off adapter, supported on a ‘lab- place the receiving vessel underneath the take-off
jack’ or clamped in position. The outlet of the take-off adapter, supported on a ‘labjack’ or clamped into
adapter must be just inside the receiving vessel so position. Turn up the water flow to give a steady
that no spillage of distillate will occur. If you clamp the stream from the condenser.
Box 16.2 How to carry out a simple distillation
1. Set up the apparatus as described in Box 16.1 and the liquid and record the temperature range of the
make sure that all the joints in the distillation system distillate.
are secured properly. 4. Remove the heat source, allowing the apparatus to
2. Slowly apply the heat source to the distillation until cool down and the remaining drops of distillate to run
the liquid is boiling gently. out of the condenser into the receiving flask.
5. Seal and label the receiving flask(s).
3. Increase the heat slowly until the liquid starts to
drip into the receiving vessel at a rate of about one 6. When cool, dismantle the apparatus, wash it out with
drop every 2 seconds. If the temperature is ‘con- an appropriate solvent and dispose of the washings
stant’ (i.e. it does not vary more than 2–3 °C), collect in the correct solvent residues bottle.

Distillation
Box 16.3 How to carry out a fractional distillation
1. Set up the apparatus as described Box 16.1 but insert 5. Increase the heat slowly and collect the intermediate
the fractionating column between the distilling flask fraction until the temperature stabilises again. Record
and the still-head. For extra stability, clamp the frac- the temperature range of the distillation.
tionating column at its top joint to support the stand
carrying the distilling flask. 6. Change the receiving flask and collect the new frac-
tion (one drop every 2 seconds) for as long as the
2. Apply the heat source slowly until the liquid begins temperature remains ‘constant’ (i.e. it does not vary
to boil gently. Then slowly increase the heat until the by more than 2–3 °C). Record the temperature of the
column is warmed up and liquid is condensing from distillation.
the thermometer bulb, but not distilling over.
7. Remove the heat source, allowing the apparatus to
3. If the temperature is constant, increase the heat cool down and the remaining drops of distillate to run
slightly until the first component distils over at a rate out of the condenser into the receiving flask.
of about one drop every 2 seconds. Collect this dis-
tillate as the first fraction, as long as the temperature 8. Seal and label the receiving flask(s).
is ‘constant’ (i.e. it does not vary more than 2–3 °C).
Record the temperature range of the distillation. 9. When cool, dismantle the apparatus, wash it out with
an appropriate solvent and dispose of the washings
4. Change the receiving flask when the temperature in the correct solvent residues bottle.
begins to rise and the rate of distillation slows.
liquid. After the first fraction has distilled, the temperature will rise and the rate
Safety Note You must check all of distillation will slow. This is an intermediate fraction containing a little of
flasks and glassware for ‘star’ cracks both components of the mixture. Finally, the temperature will become constant
and chips. If in doubt, replace. and the pure higher boiling compound will distil. The procedure for fractional
distillation is given in Box 16.3.
Key Point A successful fractional distillation relies on thermal

Working with reduced pressures – equilibration of the components in the fractionating column. You
distillation flasks and receiving flasks must allow the column to be heated by the vapours of the mix-
must always be round bottom or pear ture. Take your time!
shaped. Do not use flat-bottom or coni-
cal flasks under vacuum.
Vacuum or reduced-pressure distillation

Measuring reduced pressures – note Distillation at reduced pressure is used to distil liquids with few impurities or to
that despite SI nomenclature for pres- fractionate the components of liquid mixtures with high boiling points, which
sure, practical work usually involves would decompose if distilled at atmospheric pressure. The modifications to the
pressure measurement in mm of Hg. distillation apparatus (Fig. 16.1) required for reduced-pressure distillation are:
Atmospheric pressure is about 760 mm ●● A vacuum source: this can be a water pump (see p. 85) with an ‘anti-suck-
Hg. Pressures quoted in ‘torr’ (seen back’ trap producing a vacuum of 15–25 mm Hg, at best; or a rotary vacuum
on some old vacuum equipment) are pump (consult your instructor about its use since it is an expensive piece of
equivalent to mm Hg. equipment and easily contaminated or damaged), which will evacuate down
to 0.1 mm Hg. Two-stage dry vacuum pumps produce a vacuum of 1–5 mm
Hg, are resistant to organic and acid vapours and are easy to use.
Moving a mercury manometer – always
carry mercury manometers in a vertical
●● A manometer to measure the pressure (vacuum) in the apparatus, since the
position or the mercury may spill out.
boiling point of a liquid varies with pressure. Two types are in common use
(Fig. 16.3): the Anschutz manometer, which gives a constant reading of the

Distillation
vacuum
left-hand limb filled ‘pig’ type

with mercury receiving
adapter
receiving
to aspirator flask
to apparatus via trap
Anschutz manometer Vacustat® Fig. 16.4 ‘Pig’-type receiving flask adapter.
Fig. 16.3 Manometers for vacuum distillation.
vacuum throughout the distillation, and the Vacustat®, which is used to take
a ‘sample’ of the vacuum at a given instant. Vacustats® are very accurate and
are more often used in combination with a rotary oil pump.
●● A take-off adapter, which permits the collection of several fractions without
stopping the distillation to change the receiving flasks. A receiving adapter,
termed a ‘pig’ (Fig. 16.4), can be rotated on the end of the condenser to
collect three fractions.
●● Appropriate ‘anti-bumping’ measures: reduced-pressure distillations are
notoriously prone to ‘bumping’ (p. 87) and anti-bumping granules are inef-
Safety Note If, during the course fective. You can use a magnetic ‘flea’ in conjunction with a hot plate-stirrer
of your vacuum distillation using a and oil bath; an air bleed, made by pulling out a Pasteur pipette into a fine
micro-burner and a boiling stick, the capillary using a microburner (consult your instructor), inserting it into a
liquid stops boiling and appears ‘qui- screwcap adapter (p. 100) and placing it in the second neck of the distilling
escent’, it is about to ‘bump’ vigorously. flask. The vacuum pulls a fine stream of air into the flask and forms small
Stop heating, allow it to cool, ‘break’ bubbles which agitate the liquid during distillation; or a wooden boiling stick
the vacuum (Box 16.4), and add a new for small-scale distillations of short duration, since the vacuum will pull air
boiling stick. from the stick and agitate the liquid.
●● A three-way tap inserted between the vacuum source and the manometer, so
that you can control the vacuum applied to the distillation system.
Safety Note Do not allow air to
A typical system for reduced-pressure distillation is shown in Fig. 16.5 and the
rush into an Anschutz manometer, oth-
procedure, using a water pump, is described in Box 16.4.
erwise the mercury in the left-hand limb
of the manometer will shoot up the
tube and may break the glass at the top.
Quickfit® thermometer
fractionating column
support for
pressure tubing
water pump ‘trap’
three-way tap
air bleed manometer
with clip
Fig. 16.5 Apparatus for vacuum distillation.

Distillation
Box 16.4 How to carry out a reduced-pressure distillation using a water pump
1. Set up the distillation apparatus as described in supported with a clamp and stand, close to the ‘pig’,
Box 16.1, but use a two-necked distilling flask and to prevent strain on the glassware.
include a short fractionating column between the
top of the distilling flask and the still-head or use a
8. Slowly turn the three-way tap to evacuate the appa-
ratus. As the pressure decreases, bubbles will issue
Vigreux flask (Fig. 16.2) or use a Claisen still-head
from the air bleed or the boiling stick and low-boiling
(Fig. 16.2).
solvents such as dichloromethane or diethyl ether will
2. Connect three Quickfit® round-bottom receiving begin to boil, causing frothing in the flask. If this hap-
flasks to the receiving adapter or ‘pig’ using plastic pens, stop applying the vacuum via the three-way tap
joint clips and attach the ‘pig’ to the bottom of the and wait until the frothing has died down and then
condenser. Support the receiving flasks on a ‘labjack’ continue to lower the pressure until the three-way tap
if they are 100 mL size or larger. Rotation of the ‘pig’ is fully open.
allows three fractions to be collected without stop-
9. Check whether the air bleed is too coarse, produc-
ping the distillation – this is a major task when work-
ing large bubbles and vigorous splashing in the dis-
ing under reduced pressure.
tillation flask. If so, place a piece of pressure tubing
3. Insert the air bleed into the ‘spare’ neck of the distill- (about 2 cm long) on the end of the air bleed and
ing flask or into the lower joint on the Vigreux flask constrict the tubing with a screw clip to reduce the
or Claisen head and make sure that the tip of the air air flow through the air bleed to an acceptable rate.
bleed reaches the bottom of the distilling flask. If you
10. Slowly open the tap on the manometer, allow the
are using boiling sticks as an ‘antibumping’ precau-
mercury levels to stabilise and read the vacuum by
tion, add the sticks, making sure that they reach to the
moving the zero on the scale to the lower mercury
bottom of the flask and do not float, and put stoppers
level: the reading should be 15–25 mm or better. If
in the unused joints.
this pressure is not obtained, there must be leaks in
4. Insert a Quickfit® thermometer into the remaining the system, which usually occur at the ground-glass
‘male’ joint in the apparatus. Do not use an ordinary joints or rubber-to-glass joints. Close the tap on the
thermometer in a screwcap adapter: it may be sucked manometer, check the sealing of the joints by rotating
into the apparatus and break or crack the flask. each one in turn and also check the rubber-to-glass
joints by carefully pushing the rubber pressure tub-
5. Using thick-walled rubber pressure tubing, connect ing a little further onto the glass. Open the tap on the
the water pump (via the anti-suck-back trap) to the manometer and recheck the pressure.
three-way tap and then to the Anschutz manome-
ter. Connect another piece of pressure tubing to the 11. Gently heat the distilling flask and carry out a frac-
manometer but do not connect it to the ‘pig’ for the tional distillation as described in Box 16.3, collect-
moment. ing the fractions by rotating the ‘pig’. Remember to
record the temperature and pressure at which the
6. To check the available vacuum, turn on the water fractions distil. If the liquid in the distilling flask
pump fully, kink or seal the open pressure tubing on bumps over into the condenser, you will need to
the manometer and turn the three-way tap so that a clean out the apparatus with a solvent, evaporate off
vacuum is created in the manometer. Slowly open the the solvent to recover the chemicals and restart the
tap on the manometer and the mercury level should whole process.
rise in one of the manometer ‘arms’. When the mer-
cury has stabilised, move the scale so that the zero 12. When the distillation is complete, close the tap on
is level with the lowest mercury level, and read off the manometer and allow the apparatus to cool. Sup-
the upper mercury level. The pressure (vacuum) is port the ‘pig’ and receiving flasks (with your hand or a
the difference in levels in mm. The reading should be ‘labjack’) and slowly open the three-way tap to allow
between 10 and 20 mm. Now close the tap on the air into the system. Disconnect the pressure tubing
manometer, unkink or unseal the pressure tubing, from the ‘pig’ and place the ‘pig’ and receiving flasks
and turn the three-way tap so that air is admitted into in a safe place. Disconnect all other tubing and appa-
the system. ratus for washing or storage, turn off the water pump
and finally, very gently, open the tap on the manom-
7. Connect the pressure tubing from the manometer to eter to allow the mercury levels to equalise and then
the ‘pig’. Pressure tubing is heavy and it should be close the tap.

Distillation
oven motor
tapered joints vacuum

Ending a distillation – complete distil-
lation of a liquid is not possible because
of the need to leave a few millilitres in
heating control
the distilling flask, to prevent overheat-
temperature gauge
ing and due to the ‘hold-up’ volume of
the flask and fractionating column. Fig. 16.6 Kugelrohr distillation apparatus.
Kugelrohr distillation
Estimating the boiling point of a liquid
at reduced pressure – a useful guide This is also known as short-path distillation or bulb-to-bulb distillation. It is
is: if the pressure is halved, the boiling a procedure for reduced-pressure distillation, which almost eliminates losses
point is reduced by ∼ 10 °C. For exam- owing to the size of the apparatus, and is particularly useful for small volumes
ple, if the b.pt is 300 °C at 760 mm Hg, of liquids. The liquid sample is placed in a small round-bottom flask, con-
it will be approximately 290 °C at 380 nected to a series of bulbs (Fig. 16.6) and then heated and rotated (to prevent
mm Hg and 190 °C at ∼ 1 mm Hg. Alter- bumping) under vacuum in an electric oven. The liquid distils from the flask
natively a nomograph (Fig. 16.8) can be inside the oven to a cold bulb outside the oven. Since the distillation path is
used for a more accurate estimation. very short the length of a ground-glass joint losses are minimised. The use of
several bulbs as receiver flasks allows a small volume of mixture to be frac-
tionated by varying the temperature in the oven. The distillates can be removed
from the bulbs by either washing into another flask with a low-boiling-point
solvent or using a ‘bent’ Pasteur pipette (Fig. 16.7) since the bulbs can only
be held horizontally.
Steam distillation
This process can be used to separate water-insoluble covalent compounds from
crude reaction mixtures or to isolate volatile natural products, e.g. terpenes
from plant tissue. The crude mixture is heated with water and steam and the
steam-volatile material co-distils with the steam and is then condensed with
the water and collected. You will then need to carry out an extraction (p. 139)
Fig. 16.7 A ‘bent’ Pasteur pipette. to separate the water and the insoluble component.
The steam required for a steam distillation can be provided by an external
source, such as piped steam in the laboratory or steam generated by heating
water in a flask, which is then piped into the distilling flask. Steam is very
dangerous and the safest way to generate steam is to heat the compound with
a vast excess of water in the distilling flask: steam is generated in situ. The
equipment for steam distillation is illustrated in Fig. 16.9 and the procedure is
Key Point You must never attempt a distillation in a com-

pletely closed system. Always check that the expanding vapours
can escape.

Distillation
0.0
1
0.0
2
0.0
0. 3
0.0 04
`C 0.0 5
0.0 6
400 0.1 8
`C
0.2
700 0.3
0.4
0.6
0.
600 1.0 8
300
2
500 3
4
6
8
400 10
20
30
200 40
300 60
8
1000
200
200 300
50
7000
100 100
pressure in mm Hg
b.pt. corrected
to 760 mm Hg
observed b.pt.
Fig. 16.8 Nomograph for estimating boiling point at reduced pressure. To

use it, draw a line between the recorded pressure and the boiling point at
760 mm Hg, and then extend the line to the observed boiling point scale.
This point is the boiling point at the reduced pressure. In this example, a
liquid b.pt. of 250 °C at 760 mm Hg will boil at 118 °C at 10 mm Hg.
splash head
condenser
water for replenishing

distillation pot
clamp
water out joint clip

clamp
Claisen adapter
water in
clamp
heat source
Fig. 16.9 Apparatus for steam distillation.

Distillation
Box 16.5 How to carry out a steam distillation
1. Set up the apparatus for distillation as described in 2. Add the compound to be distilled to the flask via the
Box 16.1 with the following modifications: spare neck, half-fill the distillation flask with water,
add a large portion of ‘anti-bumping’ granules and
(a) Use a Bunsen burner with tripod and gauze as the fill the addition funnel with water.
heat source.
3. Heat the flask until steady boiling commences and
(b) Use a large three-necked flask as the distilling
distillation of an oily emulsion begins. Then con-
flask, usually 250 mL or 500 mL capacity for most
tinue the distillation adding water from the addition
laboratory procedures or a single necked flask
funnel to maintain the level of water in the distilla-
with a Claisen head (Fig. 16.9).
tion flask.
(c) Use a splash-head instead of a still-head since
there is no point in recording the distillation 4. If the distillate is a solid, it may clog and block the
temperature and it is more important not to con- condenser. If this occurs, turn off the condenser
taminate the distillate by splashing from the dis- water, take the condenser tube off the tap to let the
tillation flask. water drain out of the condenser and allow the steam
to heat up the condenser and melt the solid, which
(d) Insert an addition funnel (a separating funnel will then flow into the receiving flask. Then carefully
with a ground-glass joint on the stem) into the restore the water flow.
distilling flask and fit a stopper in the remaining
neck of the flask. 5. The distillation is complete when no more oily emul-
(e) Use a single-surface condenser: it is easier to sion is condensing (check by collecting a few drops
unblock than a double-surface condenser. of distillate in a clean, dry test tube). Turn off the heat
and allow the apparatus to cool before dismantling.
(f) Use a simple take-off adapter leading into a large
conical flask since you will be collecting a large 6. Separate the product from the water by extraction
volume of water. into a suitable organic solvent (p. 139).

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Martinez-Garcia, H. (2015) Experimental Organic Cole, Boston.
Chemistry Laboratory Manual. Elsevier, Amsterdam. Zubrick, J.W. (2015) The Organic Chem Lab Survival
Keese, R., Brandle, M.P. and Toube, T.P. (2008) Practi- Manual. A student’s guide to techniques, 10th edn. John
cal Organic Synthesis: a student’s guide. John Wiley & Wiley & Sons Ltd, Chichester.
Sons Ltd, Chichester.
Lehman, J.W. (2009) Student Lab Companion: Laboratory
Techniques for Organic Chemistry, 2nd edn. Pearson,
Harlow.

Distillation
Study exercises
16.1 An organic liquid has a boiling point of 340 °C (c) 20 mm Hg

at 760 mm Hg. Using the information on p. 150, (d) 1.0 mm Hg
estimate its approximate b.pt. at the following (e) 0.1 mm Hg
pressures:
16.2 Use the nomograph (Fig. 16.8) on p. 152 to make
(a) 190 mm Hg an accurate estimation of the boiling point of the
(b) 100 mm Hg liquid using the information in 16.1 above.

17 Reflux
Reflux is one of the most common techniques you will encounter in your chem-
When carrying out reactions using vol-
istry laboratory classes. Since many reactions between covalent compounds are
atile or dangerous chemicals or sol-
slow processes rather than instantaneous reactions, prolonged heating forces
vents – use a reflux system to keep the
the equilibrium to give an acceptable amount of product. In the reflux process,
vapours in the apparatus even though
the reactants are dissolved or suspended in a suitable solvent, the solvent is
the mixture is not heated to its boiling
boiled and then condensed so that it returns to the reaction flask. Once set up,
point.
a reaction carried out under reflux can be run for minutes, hours or even days to
promote the required reaction. The basic components for a reflux apparatus are:
Refluxing overnight – you must have ●● a reaction flask;

the approval of your instructor for these ●● a reflux condenser;
operations and complete the necessary
documentation for the night-staff. A ●● a heat source (see p. 87);
special laboratory may be available for ●● a coolant source, usually water, for the condenser.
‘overnight’ reactions.
Since the water pressure may change The procedure for setting up a simple reflux apparatus (Fig. 17.1) is given
during the night, you must fix the cool- in Box 17.1.
ant tubes in place, on the condenser
Reaction flasks
and the tap, using copper wire twisted
round the rubber tubing or plastic cable Round-bottom or pear-shaped reaction flasks are preferred, but note that stir-
ties. ring with the usual type of magnetic ‘flea’ is not possible in pear-shaped flasks.
The flasks can have multiple necks so that the apparatus can be configured for
temperature measurement, addition of solids or liquids, mechanical stirring and
inert atmosphere work (p. 166). No matter which arrangement of components
Stirring in pear-shaped flasks – special
is used, always clamp the reaction flask at the neck and keep the heaviest
triangular-shaped ‘fleas’ are now avail-
components (such as an addition funnel containing another chemical) vertically
able for the conical-base tubes used for
above the flask. A condenser will still function at 30° from vertical and it is
evaporation and microscale reactions,
not very heavy.
but they are not yet common in the
undergraduate laboratory. Condensers
For general-purpose work, these are usually single-surface or double-surface
types (Fig. 17.2): the double-surface condenser is used for low-boiling
point solvents such as dichloromethane, diethyl ether or light petroleum
(b.pt. 40–60 °C).
water out
condenser
thermometer water in
clamp
magnetic
stirrer bars
heating bath
(oil etc.)
magnetic stirrer/
hot plate single double coil
surface surface
Fig. 17.1 Apparatus for simple reflux. Fig. 17.2 Condensers.

Reflux
Box 17.1 How to set up a simple reflux apparatus
1. Choose a round-bottom or pear-shaped Quickfit® 5. Fit the rubber tubing to the condenser; the lower tube
flask of appropriate size so that it will be no more is connected to the water tap so that the water flows
than half-full and clamp it to a support stand. up the condenser for the most efficient cooling.
2. Select the appropriate heat source and anti-bumping 6. Lightly grease the joint on the condenser and place it
protection, and adjust the height of the flask so that in the flask, rotating it to ensure a good seal. Do not
the heat source can be removed easily if something clamp the condenser – gravity will keep it in place –
goes wrong. but make sure that it is vertical. You may need to
3. Choose a condenser of appropriate size and type so move the set-up quickly away from the heat source
that it will condense the volume of vapour formed in and it is easier to manipulate if only one clamp, on the
the reflux process. For example, do not use a small neck of the flask, is present.
condenser with a 14/23 joint on a 250 mL flask, since
it will not cope with the volume of vapour to be con- 7. Tidy the coolant pipes behind the clamp ensuring that
densed, or a large condenser on a small flask, where there are no kinks, the pipes are not touching the heat
all the solvent may be converted into vapour before source and the outlet pipe is positioned in the drain or
condensation occurs. sink. Turn on the water gently to check for leaks and, if
all is correct, turn up the water to give a steady flow.
4. Add the liquids/solids and the solvent to the flask,
using the solids funnel, paper funnel or stemmed fil- 8. Apply enough heat to bring the liquid to the boil,
ter funnel (p. 80) so that you do not get chemicals on check that the solvent is refluxing at a steady rate
the inside of the ground-glass joint. If you do contami- and that the vapour is condensing no higher than
nate the joint, it may not allow the condenser to ‘seat’ one-third of the length of the condenser. The appara-
properly and hot solvent vapour will escape into the tus can then be left, with regular monitoring, for the
atmosphere. Wipe the joint clean with tissue. reaction to proceed.
outer rubber gasket
to coolant
pipe
Rubber tubing for coolant water is attached to the condenser in two ways:
1. If the condenser has glass inlet and outlet pipes with a ‘knuckle’, wet the
inside of the rubber tube with a little water and slide it onto the pipe and
past the ‘knuckle’. The rubber tubing must be a tight fit otherwise it may
slide off over a period of time.
inner rubber gasket 2. Modern condensers have plastic adapters, which can be attached to the tub-
ing and then screwed on the threaded inlet and outlet pipes. Slide the rub-
Fig. 17.3 Plastic connectors for condensers. ber tubing onto the moistened ‘pipe’ on the adapter, and then screw the
adapter onto the condenser. You must ensure that the adapter has a rubber
Refluxing anhydrous reactions – if it
gasket on the inside of the threaded portion (Fig. 17.3), otherwise it will
is necessary to exclude atmospheric
leak water at the condenser inlet or outlet.
water or oxygen or carbon dioxide,
guard tubes are ineffective and the
reaction must be carried out under an
Key Point You should always attach rubber tubing to a con-
inert atmosphere (p. 166).
denser before fitting it to the apparatus.
Safety Note Always use a fresh

guard tube because the action of water Drying tubes
on the drying agent may form a solid
‘cake’ and seal the guard tube. This will
Water can get into your reaction by condensation from the atmosphere or by
act as a ‘stopper’ and pressurise the
condensation of the steam produced in a water bath. To exclude water, you
reflux apparatus when you begin heating
should fit a guard tube containing a solid drying agent such as anhydrous cal-
and may cause an explosion. Do not pack
cium chloride or calcium sulphate to the top of the condenser. A typical guard
the tube too tightly with drying agent.
tube is shown in Fig. 17.4; use a coarse-sized drying agent rather than a fine
powder, which would ‘cake’ very quickly as it absorbs moisture.

Reflux
cotton wool Reflux with addition of chemicals

Instead of stopping the reaction and opening the apparatus, you can put in the
new chemicals using an addition or ‘dropping’ funnel. Addition funnels are
separatory funnels (see p. 141) with a ground-glass joint on the stem, and there
are two types (Fig. 17.5):
CaCl2 lumps 1. Addition funnels: when you add a liquid or solution from the funnel to
the reaction flask, you must take out the stopper, otherwise a vacuum is
Fig. 17.4 A CaCl2 guard tube.
formed and the liquid does not flow out (see p. 142). This is a disadvan-
tage when using compounds with irritating or toxic vapours or which are
Safety Note Never attempt to use air sensitive. The simplest solution to this problem is to fit a guard tube,
a pressure-equalising dropping funnel instead of a stopper, to the addition funnel and then the liquid or solution
as a separatory funnel because, when will flow easily into the reaction flask.
you invert the funnel, the solvent will 2. Pressure-equalising dropping funnels: these have a side arm linking the
flow down the side arm, past the tap reservoir of the funnel to the inlet stem below the tap. The pressures
and onto you or the laboratory bench! in the reservoir and the reaction flask are equal, and liquid will flow
into the reaction flask even when the stopper is in place in the funnel.
Pressure-equalising dropping funnels are very expensive and are normally
only used for inert atmosphere reactions (p. 167).
Box 17.2 How to set up the apparatus for reflux with mechanical stirring
1. Clamp a multineck flask to a heavy support stand well. You can increase the speed to check for vibra-
at a height where you can put the heating source tion and ‘whip’ and then turn the speed down to
(mantle or oil bath) underneath, on a ‘labjack’. zero. If the stirrer is reluctant to stir or seems ‘stiff’,
switch off the power and readjust the apparatus.
2. Slide the stirring rod through the stirrer gland, add
the stirrer paddle and fit into the clamped joint of 7. If the glass stirring rod ‘slips’ in the rubber tubing,
the flask. Lift the stirrer rod so that the paddle is not tighten the tubing round the glass rod with twisted
touching the bottom of the flask. copper wire or a plastic cable tie.
3. Slide a piece of rubber pressure tubing (about 40 8. Add the chemicals to the flask (stemmed funnel)
mm long) half-way onto the drive shaft of the stir- through one of the other necks, grease and fit a reflux
rer motor and fix the stirrer motor onto the sup- condenser and addition funnel (if appropriate). Start
port stand about 3 mm above the top end of the the motor and increase the speed to give a steady
stirrer rod. stirring rate.
4. Very carefully line up the centre of the pressure tub- 9. Raise the heat source under the flask and adjust the
ing on the motor with the centre of the stirring rod power to give steady boiling.
and ensure that the drive shaft, stirring rod, stirrer 10. When the reaction is complete, remove the heat
gland and flask are aligned and vertical: look at the source and allow the apparatus to cool to room
set-up from several different angles and adjust the temperature. Switch off the stirrer and disconnect
clamps as appropriate until you are sure. from the mains and remove the condenser and addi-
tion funnel. Carefully lower the reaction flask about
5. Lift the stirring rod and slide it into the rubber tub-
40 mm (the stirrer paddle will be lifted from the bot-
ing, about 10 mm, and then raise the flask so that the
tom of the flask by 40 mm), cut the copper wire or
stirrer paddle is about 2 mm above the bottom of the
plastic tie holding the top of the stirrer rod in the rub-
flask. Finally tighten all the clamps firmly.
ber tube and carefully ease the stirrer rod out of the
6. Make sure that the stirrer motor speed control is rubber connection. Unclamp the stirrer motor and put
set to the minimum speed or zero. Switch on the it aside. You can now remove the stirrer and stirrer
power and turn the speed control to the lowest set- gland from the reaction flask, rinsing the paddle into
ting. If the stirrer turns smoothly and slowly, all is the flask with a little fresh solvent.

Reflux
The addition of dry solids to refluxing reactions requires special equipment

Adding a chemical to a refluxing reac-
and you should refer to the appropriate texts (Errington, 1997; Furniss et al., 1991);
tion – many reactions are exothermic,
Harwood et al., 2000). The simplest approach is to dissolve the solid in a small
so you should add the new chemical
amount of the solvent being used and add it as a solution, using an addition funnel.
to the reaction at such a rate that the
height of the refluxing vapour in the Reflux with mechanical stirring
condenser does not change much.
When a magnetic stirrer is unsuitable, e.g. in reactions involving viscous liquids
or mixtures of solids and liquids, a mechanical stirrer must be used. A mechan-
ical or overhead stirrer system comprises:
1. An electric variable speed motor connected to the mains supply.
2. A flexible connector, usually a short length of rubber tubing.
guard tube
water out
addition
stopper funnel
condenser
pressure
water out equalising water in
(a) (b) funnel
clamp
clamp clamp
Fig. 17.5 Adding chemicals to a
condenser
reflux apparatus: (a) addition funnel; Claisen
(b) pressure-equalising funnel. adapter
clamp clamp
water in
heating
bath
two-neck flask
magnetic
boiling stones
stirrer bars
heating source
Heat sources – in cases where multi-
ple clamps or mechanical stirring are magnetic stirrer/
hot plate
used, the heat source must be mounted
on a ‘labjack’ so that the heat source
(a) (b)
can be removed quickly if something
should go wrong.
water out
condenser
addition
funnel
sealed stirrer
guide
clamp
water in
three-neck flask
heat source
(bath, heating mantle)
(c)
Fig. 17.6 Apparatus arrangements for reflux with addition: (a) heating a
reaction mixture to reflux during the addition of a reagent; (b) heating a
stirred reaction mixture during the addition of a reagent; (c) heating and
overhead stirring of a reaction mixture during the addition of a reagent.

Reflux
3. A stirrer gland, which fits into the top ground-glass joint of the flask acting
as a seal for the refluxing vapour and a guide for the stirrer shaft. There are
several types of gland available, but the best is now made from Teflon® and
needs no lubrication. If this is not available, a screwcap adapter in which
the rubber sealing ring has been lubricated with a touch of silicone oil can
be used. Note that the stirrer shaft should rotate in the adapter, not the
adapter in the joint of the flask, so do not tighten the screwcap too much.
4. A Teflon® paddle which swivels on the end of the stirrer shaft so that it can
pass through the ground-glass joint on the top of the flask.
Setting up the apparatus for reflux with mechanical stirring is a precision
task and is described in Box 17.2. The major problems encountered are:
●● The weight of the stirrer motor high up on the support stand: use a large sup-
port stand with a heavy base or use a support framework, which is screwed
to the laboratory bench. Besides the motor’s weight, the torque of the motor
can cause ‘whipping’ in the support stand.
●● The motor, stirrer gland, stirrer shaft and reaction flask must be absolutely
vertical and concentric, otherwise the glass stirrer shaft will snap.
●● Since there will always be a little sideways movement when the stirrer is
operating, the flask and the motor should be clamped in position on the same
stand. Condensers and addition funnels should NOT be clamped.
A selection of configurations, suitable for most reflux procedures is shown
in Fig. 17.6.
Text references
Errington, R.J. (1997) Advanced Practical Inorganic and Isac-Garcia, J., Dobado, J.A., Calvo-Flores, F.G. and
Metalorganic Chemistry. Blackie Academic and Profes- Martinez-Garcia, H. (2015) Experimental Organic
sional, London. Chemistry-Laboratory Manual. Elsevier, Amsterdam.
Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch- Lehman, J.W. (2009) Student Lab Manual Laboratory
ell, A.R. (1991) Vogel’s Textbook of Practical Organic Techniques for Organic Chemistry, 2nd edn. Pearson,
Chemistry, 5th edn. Longman, Harlow, Essex. Harlow.
Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper- Leonard, J., Lygo, B. and Procter, G. (2013) Advanced
imental Organic Chemistry, 2nd edn. Blackwell Science Practical Organic Chemistry, 3rd edn. CRC Press, Boca
Ltd, Oxford. Raton.

Keese, R., Brandle, M.P. and Toube, T.P. (2008) Practical Suib, S.L. and Tanaka, J. (1999) Experimental Methods in
Organic Synthesis: A Student’s Guide. John Wiley & Inorganic Chemistry. Prentice Hall, Harlow, Essex.
Sons Ltd, Chichester. Williamson, K.L. and Masters, K.M. (2010) Macroscale
Loewenthal, H.J.E. and Zass, E. (1992) A Guide for the and Microscale Organic Experiments, 6th edn. Brooks
Perplexed Organic Experimentalist, 2nd edn. John Cole, Boston.
Wiley & Sons Ltd, Chichester. Zubrick, J.W. (2015) The Organic Chem Lab Survival
Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical Manual. A student’s guide to techniques, 10th edn. John
Organic Chemistry. Chapman & Hall, London. Wiley & Sons Ltd, Chichester.
Smart, L. (ed.) (2002) Separation, Purification and Iden-
tification – The Molecular World. Open University and
Royal Society of Chemistry, Cambridge.

Reflux
Study exercise
17.1 You are to carry out the experiment to convert Do a Hazard Assessment of the materials
cyclohexene into 1,2-dibromo-cyclohexane involved in the reaction and decide on the appa-
by the dropwise addition of liquid bromine to ratus you will use for the synthesis.
cyclohexene at room temperature followed by
heating to 100 °C for 30 minutes.

18 Evaporation
Evaporation is the process in which the solvent of a solution is converted into

Evaporation – the purpose of evap-
a vapour to leave a solid or liquid solute. There are many applications of evap-
oration is to remove the solvent from
oration, ranging from the slow evaporation of water from a solution of an ionic
a solution. Purification or separation
compound to leave hydrated crystals, e.g. crystallisation of CuSO4.5H2O, to
of the components of the solute is by
the evaporation of large volumes of low-boiling-point solvent under reduced
other means, such as recrystallisation
pressure in the extraction of an organic compound. Since crystallisation by
or distillation.
evaporation is a specific technique for a relatively small range of compounds,
the term evaporation is generally interpreted as the removal of the solvent from
a solution.
Evaporation of solvents
There are three commonly used techniques for solvent evaporation:
Safety Note Clamp the conical
flask in position on the steam bath. 1. Evaporation from open flasks on a steam bath.
There is particular risk of the flask
2. Rotary film evaporation.
falling over when using small conical
flasks ( 6 100 mL) and a glass rod. 3. Gas ‘blow-down’.
All these techniques have advantages and disadvantages. Where your
experimental protocol may simply state ‘the solvent is evaporated off’, you
should select the most appropriate procedure based on:
Using ‘rovaps’ – these are communal

●● the volume of solvent to be removed;
so make sure that the ‘rovap’ is clean ●● the amount of solute in solution;
before you use it and clean it up after
use. Empty the solvent collection flask
●● the relative boiling points of the solvent and solute;
into the appropriate waste solvent ●● the next step in the experimental procedure.
bottles.
Evaporation from open flasks
This is useful for evaporating small volumes (∼ 25 mL) of low-boiling-point
solvents (6 70 °C) from solutions containing a solute which has a boiling point
above 110 °C. Place the solution in a beaker or conical flask, containing a glass
rod or boiling stick, onto a steam bath (maximum temperature achievable is
100 °C) in a fume cupboard and evaporate the solvent until boiling ceases. The
obvious advantage is simplicity; the disadvantages include environmental con-
cerns of release of solvents in the atmosphere, contamination of the solvent by
condensed steam and incomplete solvent removal due to the ‘hold-up’ volume
of the flask.
Rotary film evaporation
This method, which is also known as rotary evaporation or ‘rovap’, is the tech-
nique of choice for the removal of large volumes of volatile solvents from
solutions, e.g. from extractions and column chromatography (p. 263). Rotary
evaporators are now standard communal pieces of equipment in the laboratory
and the operating principle is that of a reduced-pressure distillation except that
the evaporation flask can be rotated. This rotation reduces the risk of ‘bump-
ing’, inherent in all reduced-pressure distillations, and spreads the solution in
a thin film on the walls of the flask. This effectively increases the surface area
of the solution and increases the rate of evaporation, which is further enhanced
by the use of a vacuum.

Evaporation
Box 18.1 How to use a rotary film evaporator
1. Check that the apparatus is ready for use by ensuring system is an electric motor controlled by an ‘up-down’
that: pressure switch, but on older apparatus the lifting
device is either (i) a manual handle with a trigger,
(a) The receiving flask is clipped in place and is which is pulled to lift and released to lock it in place,
empty. or (ii) a lever with a twist-grip on the end. To operate
(b) You have available the correct-size ground-glass the latter mechanism, twist the grip anti-clockwise
joint adapters to connect your flask to the rotating to release the ‘lock’, pull the lever down to raise the
‘barrel’ protruding from the motor of the evapo- apparatus and twist the grip clockwise to lock it in
rator. Many rotary film evaporators have an ‘odd’ place.
joint size, usually 29/32, which is not common to
5. Attach the flask to the barrel and put plastic joint clips
the routine ground-glass flasks used in the lab-
on all the joints, while supporting the flask with your
oratory. Alternatively, special bulb-shaped flasks
hand. If the weight of the flask and contents ‘springs’
with 29/32 joints may be available.
any of the joints, it is too heavy: replace it with a
(c) The rotating barrel is ‘clipped’ in place in the smaller flask or remove some of the solution. You
motor, by pulling it gently. Someone may have must not rely on the power of the vacuum to hold
had to clean out and reassemble the ‘rovap’ and, your flask in place.
if the barrel is not ‘clipped’ in place, it will slide
out when you attach your flask. If the barrel slides 6. Turn on the motor, slowly increasing the speed to the
out of the motor when you pull it, consult your maximum.
instructor. 7. Close the vacuum inlet adapter slowly until it is fully
(d) The rotating barrel is clean and dry. shut and observe the flask for a few seconds. If boil-
(e) Water is flowing steadily through the condenser. ing occurs (liquid is condensed to the receiver flask)
If it is not, adjust the water tap. continue until boiling stops and then lower the flask
so that it just touches the surface of the water, lock
(f) The temperature of the water in the water bath is
it in place and boiling should recommence. As the
about 20 °C below the boiling point of the solvent
volume of solution decreases, you may need to lower
to be removed.
the flask further into the water bath until all the sol-
vent has been evaporated. If a white coating of frost
2. Open the vacuum inlet adapter at the top of the con- forms on the flask evaporation may stop, because the
denser, and turn the water pump to maximum (p. 85). flask is too cold: lower the flask into the water bath to
3. Fill the rotating flask half-full or less, using a stemmed warm it and evaporation should begin again.
filter funnel. You must not contaminate the joint of 8. When evaporation is complete, raise the flask from
the rotating flask with solute, which will be deposited the bath, switch the motor off, open the vacuum inlet
there during evaporation, since you may not be able tap to allow air into the system and allow the flask to
to remove the flask after evaporation. If the flask is cool. Support the flask with your hand, take off the
too full it may ‘bump’, sending solution up into the plastic joint clips, put the flask on one side and only
condenser and receiver. You will then have to disman- then turn off the water pump. Turn off the water sup-
tle and clean out the equipment with an appropriate ply to the condenser.
solvent to recover your compound.
9. Unclip the receiving flask, dispose of the solvent into
4. Raise the apparatus using the lifting mechanism so a waste solvent bottle and reattach the receiving flask
that, when the flask is attached, it is not touching the to the apparatus.
water in the bath. On modern equipment the lifting
Key Point When using a ‘rovap’ you must check that the
reduced-pressure boiling point of the solute you are trying to
isolate is below the temperature of the water bath.
There are many variations in the details of the form of rotary film evaporators
and a typical assembly is illustrated in Fig. 18.1. A general guide to the use of
a rotary evaporator is given in Box 18.1.

Evaporation
Safety Notes When using rotary (a)
film evaporators you should take note

of the following: vacuum inlet adapter
●● Never use flat-bottom flasks or conical
flasks under reduced pressure (p. 148).
●● Always check your flask for ‘star’
motor
cracks.
barrel
●● Always make sure that your solution
has cooled to room temperature
before you begin, otherwise it may
boil vigorously and ‘bump’ when you
apply the vacuum, before it is low- receiving flask evaporation flask
ered into the water bath.
●● Do not rush to lower the flask into (a)
(b)
the water bath: wait to see what hap-
pens to the extent of evaporation at Fig. 18.1 (a) Typical examples of a rotary evaporator; (b) exploded view of
vacuum inlet adapter
room temperature. glassware.
●● Always have the water bath just warm,
not hot, at the start of the procedure. motor
If the water bath is too hot, allow it to If it is necessary to evaporate volumes of solvent greater than the capacity
cool or add cold water or ice. of the rotating flask, you can carry out the process batch-wise involving several
barrel
●● Check that all joints are ‘sealed’ and separate evaporations or the rotary evaporator can be modified for continuous
that the water pump is producing a evaporation (Fig. 18.2). A thin Teflon® tube is attached to the vacuum inlet
vacuum: it will change ‘note’ as the adapter so that it feeds down the condenser into the ‘barrel’ and another glass
vacuum is produced, when it is work- tube, dipping into the solution to be evaporated, is connected to the air inlet
ing properly. If there is no vacuum, receiving flask evaporation flask
on the vacuum adapter. Once the rotary evaporator is operating, the tap on the
the solution may not boil and you
vacuum adapter(b)is opened a little. Solution is drawn up by the vacuum, runs
will overheat it in trying to promote
into the rotating flask and is evaporated. Careful control of the tap allows a
evaporation. The joints may sud-
denly seal and the solution will then constant volume of solution to be sucked into the rotating flask and evaporated
boil vigorously under the reduced without overfilling it.
pressure and will ‘bump’ into the
condenser and receiving flask.
Transferring viscous liquids – it is often

difficult to transfer small amounts of
viscous liquids from a ‘rovap’ flask to a
small sample tube. Dissolve the liquid
in a small amount of dry solvent, trans-
fer a little of this solution (1 or 2 mL) to
a suitable small tube and ‘blow off’ the Fig. 18.2 The procedure for continuous solvent removal using a rotary
solvent with nitrogen. evaporator.
Gas ‘blow-down’
Safety Note Make sure that you This procedure is useful for removing very small volumes of solvents (about
do not put the tip of the Pasteur pipette
2 mL) from solutes by blowing a stream of nitrogen over the surface of the
too close to the liquid surface or you
solution, while warming the solution gently. The main application of the gas
will blow the liquid out of the tube! Test
blow-down is in the isolation of small amounts of solute from rotary evapora-
the flow first.
tion or small-scale liquid–liquid extraction, for further analysis by instrumental

Evaporation
air or nitrogen techniques, where the sample size may be 20 mg or less: for example, infra-
red spectroscopy (p. 320), NMR spectroscopy (p. 334), gas chromatography
(p. 255) or liquid chromatography (p. 260).
The simplest system for evaporation by gas blow-down is shown in
Fig. 18.3. A Pasteur pipette is connected by a flexible tube to a cylinder
of nitrogen, which has a gas blow-off safety system (p. 166). The sample
Pasteur pipette is placed in a special tube with a conical base, such as a ReactiVial®. Hold
the Pasteur pipette and direct a gentle stream of nitrogen towards the side
of the tube so that it flows over the surface of the liquid. As the solvent
evaporates, the liquid and tube will cool and may condense atmospheric
water into the tube. To prevent condensation, clamp the tube in a warm
sand bath or above a closed steam bath or in the hole of a purpose-designed
aluminium heating block. The following points should be noted when using
a gas blow-down system:
●● Always carry out the operation in a fume cupboard.
●● The solute should have negligible vapour pressure at room temperature, oth-
erwise it may co-evaporate with the solvent.
●● Do not heat the solution to boiling. Only apply enough heat to prevent con-
densation of atmospheric water vapour.
Heating medium
Fig. 18.3 Gas ‘blow-down’ evaporation.

Errington, R.J. (1997) Advanced Practical Inorganic and Leonard, J., Lygo, B. and Procter, G. (2013) Advanced
Metalorganic Chemistry. Blackie Academic and Profes- Practical Organic Chemistry, 3rd edn. CRC Press, Boca
sional, London. Raton.
Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatch- Loewenthal, H.J.E. and Zass, E. (1992) A Guide for the
ell, A.R. (1991) Vogel’s Textbook of Practical Organic Perplexed Organic Experimentalist, 2nd edn. John
Chemistry, 5th edn. Longman, Harlow, Essex. Wiley & Sons Ltd, Chichester.
Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper- Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical
imental Organic Chemistry, 2nd edn. Blackwell Science Organic Chemistry. Chapman Hall, London.
Ltd, Oxford. Smart, L. (ed.) (2002) Separation, Purification and Iden-
Isac-Garcia, J., Dobado, J.A., Calvo-Flores, F.G. and Mar- tification – The Molecular World. Open University and
tinez-Garcia, H. (2015) Experimental Organic Chemis- Royal Society of Chemistry, Cambridge.
try. Laboratory Manual. Elsevier, Amsterdam. Suib, S.L. and Tanaka, J. (1999) Experimental Methods in
Keese, R., Brandle, M.P. and Toube, T.P. (2008) Practical Inorganic Chemistry. Prentice Hall, Harlow, Essex.
Organic Synthesis: A Student’s Guide. John Wiley & Williamson, K.L. and Masters, K.M. (2010) Macroscale
Sons Ltd, Chichester. and Microscale Organic Experiments, 6th edn. Brooks
Lehman, J.W. (2009) Student Lab Companion: Laboratory Cole, Boston.
Techniques For Organic Chemistry, 2nd edn. Pearson. Zubrick, J.W. (2015) The Organic Chem Lab Survival
Harlow. Manual. A student’s guide to techniques, 10th edn. John

Evaporation
Study exercises
18.1 Decide on the most appropriate technique for (b) A solution (500 mL) of naphthalene
removal of solvent by evaporation in the follow- (m.pt. 80 °C) in ethoxyethane (b.pt. 35 °C).
ing examples: (c) A solution (2 mL) of an unknown alcohol (b.pt.
7 150 °C) in ethyl ethanoate (b.pt. 78 °C).
(a) A solution (15 mL) of nitrobenzene (b.pt.
210 °C) in dichloromethane (b.pt. 44 °C).

19 Inert atmosphere methods
During your laboratory work you may need to carry out reactions using chem-
Inert atmosphere reactions – these
icals which are described as air sensitive or moisture sensitive. These com-
should be done in the fume cupboard,
pounds may react with water, oxygen, carbon dioxide and even nitrogen (e.g.
since most of the solvents and reagents
lithium metal). The most sensitive chemicals may require special apparatus
used are flammable and/or toxic.
such as glove boxes or vacuum line equipment and you should consult the
appropriate specialist literature (Errington, 1997, p. 56). On the other hand,
many reactions involving some air-sensitive reagents (e.g. organolithium com-
Nitrogen atmospheres – note that lith- pounds or hydride reducing agent) can be done on a small scale using standard
ium metal reacts slowly with nitrogen. glassware with appropriate modifications. For simple apparatus for inert atmos-
phere reactions, the basic requirements are:
Use of argon – reactions carried out

●● Inert atmosphere, usually nitrogen or argon, piped into the apparatus. Nitro-
under argon can be opened to the
gen is the most commonly used inert gas, whereas argon is more expensive
atmosphere briefly (∼ 5 s), for the addi-
but does have the advantage that it is denser than nitrogen and is not lost
tion of other chemicals, without degra-
from the apparatus as quickly. The inert atmosphere is maintained in the
dation of the inert atmosphere.
apparatus by the use of a ‘bubbler’ (see p. 167) on one of the outlets from
the glassware – all other outlets must be stoppered or capped with septa.
●● Appropriate glassware for the experiment (see Chapter 17) which must
low-pressure
gauge be dry.
●● Dry solvents and chemicals (p. 168).
high-pressure ●● Syringe techniques for dispensing and transferring chemicals to the appa-
bursting valve gauge (cylinder) ratus (p. 168).
outlet tap
diaphragm
Inert atmosphere
regulator control
threaded bullet adapter The source of the inert atmosphere is usually a cylinder of nitrogen or argon gas
to fit into cylinder under pressure, which should be placed as close to the apparatus as possible to
increase outlet
pressure avoid long ‘runs’ of connecting rubber tubing.
cylinder valve
Gas cylinders
gas cylinder
The gas flow rate from the cylinder is controlled by the cylinder head regu-
lating valve (Fig. 19.1). Before you start make sure that the regulator outlet
tap is off (turn anti-clockwise until it feels ‘free’) and then open the valve to
Fig. 19.1 The diaphragm pressure regulator. the cylinder with the cylinder spanner (turn anti-clockwise) and the cylinder
pressure should be indicated on the right-hand pressure dial. Switch on the
Safety Note Gas cylinders must gas at the regulator (turn slowly clockwise) until there is a reading on the
be supported safely either by clamping
left-hand dial. Use the minimum pressure required to provide a steady flow
to the bench using a special cylinder
of gas. The gas flow rate from the regulator can be controlled further by a
clamp or in a cylinder trolley. needle valve on the regulator outlet, if one is fitted. To switch off, reverse the
instructions above.
Connection to the apparatus

Safety Note Hydrogen gas cyl-
Use clean, dry, thin-walled rubber tubing and special adapters with ground-
inders have special cylinder head
glass joints to connect the tubing to your reaction flask or to the inlet pipe of
regulators with a ‘left-hand thread’
a ‘bubbler’. Where a single cylinder supplies several outlets, e.g. in a fume
(tightened by turning anti-clockwise).
All ‘left-hand thread’ fittings are identi-
cupboard, the gas flow rate may change markedly when someone turns off one
fied by ‘notches’ cut into the fixing nuts
of the outlets, resulting in an increase in gas pressure to your equipment. You
(Fig. 19.2)
should, therefore, fit a gas ‘blow-off’ valve between your gas supply and the
apparatus. To do this, fit a glass tube ‘T-piece’ in the gas line to your apparatus

Inert atmosphere methods
and connect it to a Dreschel bottle containing mineral oil to a depth a little more
than that of the mineral oil in the ‘bubbler’. If there is a sudden upward change
in gas pressure, the gas will be vented through the Dreschel bottle, as well as
through your apparatus.
Fig. 19.2 A ‘left-hand’ thread, fitting.
Gas ‘bubblers’
The exit of the inert gas from the apparatus must be protected by a gas ‘bub-
Inert gas flow rate – only low flow bler’. The ‘bubbler’ allows you to monitor the flow of inert gas through the
rates are required to provide an inert apparatus and prevents the entry of air into the apparatus. Several designs of gas
atmosphere, once the apparatus has ‘bubbler’ are available (Fig. 19.3) and it is usual to connect the ‘bubbler’ to the
been ‘swept’ with the gas. apparatus at the top of the condenser. You should make sure that the ‘bubbler’
contains enough mineral oil to create a seal from the atmosphere and that it has
a bulb above the mineral oil to collect any mineral oil, which could be sucked
Using a gas ‘bubbler’ – this ensures
back into your apparatus if there is a sudden contraction in the volume of inert
that your apparatus is not a sealed sys-
gas in the apparatus. Such changes in volume will occur if you suddenly cool
tem, which will pressurise as you intro-
the apparatus without increasing the gas flow to compensate for the resulting
duce the inert gas.
reduction in inert atmosphere volume.
seal or
inert Apparatus
gas in
Depending upon the type of reaction to be carried out, one of the assemblies
shown on p. 180 may be used, with additional modifications for inert gas inlet
and outlet. You should consider very carefully what is required: heating or cool-
ing, magnetic or mechanical stirring, temperature measurement, etc. The gas
inert
gas out inlet can be either directly into the reaction flask or into the inlet arm of the gas
‘bubbler’ – it depends on the number of ‘necks’ available on the reaction flask.
inert gas out

Drying glassware
All equipment to be used should be dried (e.g. in an oven overnight at 125 °C –
inert
gas in do not forget to remove all plastic or Teflon® components before placing the
glassware in the oven). After drying, the apparatus should be greased, assem-
inert bled hot, using heat-resistant gloves as protection, and allowed to cool with
gas out
the inert gas flowing rapidly through it. Once cool, the water connections for
the condenser should be fitted – screw-on water connectors (p. 156) are most
useful in this context.
Addition of chemicals
Chemicals should be added to the reaction flask using a pressure-equalising
dropping funnel (p. 158). Liquids and solid compounds are best added as solu-
inert gas in
tions in the solvent used in the reaction. If the solid is insoluble, a little solvent
Fig. 19.3 Typical ‘bubblers’. should be added to the reaction flask, the ‘bubbler’ outlet sealed, a stopper
to the flask opened and the gas flow rate increased. The solid can then be
added from a wide-stemmed filter funnel, protected by the inert gas and solvent
Guard tubes – these absorb relatively
vapour, so that air does not enter the apparatus. Then, simultaneously unseal
little atmospheric water and do not
the ‘bubbler’ and restopper the flask, and then turn down the gas flow. Alter-
absorb oxygen and carbon dioxide.
natively a solids addition tube (Errington, 1997; Furniss et al., 1991; Harwood
Always use a gas ‘bubbler’ to protect
et al., 2000) can be used.
the exit from your apparatus.
Solvents and chemicals

All chemicals and solvents used in inert atmosphere reactions must be dry.
Most of these materials provided by suppliers are not dry enough, even solvents
which you consider to be immiscible with water, and may contain enough

moisture to hinder the reaction or reduce the yield of your product. Therefore
Dry solvents – the term also implic-
you must ensure that all chemicals to be used in the process have been dried to
itly means carbon dioxide-free and
the appropriate levels, as described below.
oxygen-free.
Solid chemicals
These should be dried by the methods outlined on p. 95. The most common
approach is to dry the chemical in an oven and then allow it to cool in a vacuum
desiccator (p. 96). Techniques for extremely air-sensitive solids can be found
in the specialist literature.
Liquid chemicals
All liquids should be dried by a method appropriate to the amount of water they
may contain (p. 97). Generally, the liquid should be dried with a solid drying
agent (p. 97) which does not react with the chemical (consult the appropriate
literature or your instructor), filtered, distilled (p. 145), then stored over molec-
ular sieves (p. 97) in a bottle capped by a septum and redistilled before use.
Alternatively, the liquid can be dissolved in a solvent, the solution dried (p. 97),
the solvent removed by evaporation (p. 161) and the liquid distilled and stored
as described above.
Solvents
The solvent will have the greatest volume in your reaction and it must be dry.
Grignard reactions are relatively
Most laboratory-grade solvents, as supplied by manufacturers, contain varying
tolerant – dry solvent can be added
amounts of water and therefore must be dried by the appropriate method before
to the apparatus using an oven-dried
use. If you are required to dry the solvent, you should consult the literature
measuring cylinder.
(Errington, 1997; Furniss et al., 1991; Harwood et al., 2000).
Some manufacturers supply dry solvents in 2.5 L quantities for inert
atmosphere reactions and HPLC (p. 265). These solvents are relatively expen-
sive but may be economic in terms of time and expense if one-off reactions
are required. However, such solvents should be treated with suspicion if the
containers are less than half-full, since air and moisture may have been allowed
into the container by previous users. If you have any doubts, dry the solvent.
Syringe techniques
Many air-sensitive chemicals are supplied as solutions in nitrogen-filled bot-
tles, which are sealed by a septum, and small volumes (up to 25 mL) of these
solutions are best transferred to the apparatus using glass syringes. Similarly,
air-sensitive liquids can be added to the reaction using a syringe.
Key Point When removing air-sensitive reagents from

nitrogen-filled bottles, you must replace the volume of liquid
removed with inert gas (nitrogen) from a gas cylinder or bal-
loon, via a needle, otherwise air (water, oxygen and carbon
dioxide) will be pulled into the bottle as a result of the vacuum
you have created.
Syringes
Glass, gas-tight syringes with a Luer lock fitting are the most versatile type of
syringe and they come in a range of sizes. The Luer lock enables the stainless
steel needle to be locked in place on the end of the syringe so that there is

no danger of the needle dropping off the syringe during the transfer process
Using syringes – always ensure that
(Fig. 19.4). Variations in syringe types include those with Teflon®-tipped pis-
the syringe piston is the correct one for
tons (plungers), which are somewhat more expensive.
your syringe, since the components of
Before using a syringe, always check that it is working by sucking up a
the syringe are always separated for
little of the solvent to be used, ensuring that air is not sucked into the syringe
drying.
either via the Luer lock or down a gap between the syringe and piston. If all
is correct, disassemble the syringe and needle, dry in an oven at 120 °C (not if
Teflon® tipped) and allow to cool in a desiccator. Once you have transferred the
air-sensitive reagent, you must clean out the syringe and needle immediately, by
glass barrel glass piston
the appropriate method, as air will get into the needle and syringe and decom-
pose the reagent causing the syringe to jam or the needle to block.
Needle-to-tubing connectors
glass barrel Teflon® tip solid piston
These adapters allow a Luer lock syringe needle to be connected to rubber
tubing (Fig. 19.5). An inert gas supply can then be piped, via the needle, into
Teflon® seal
a bottle to allow the transfer of large volumes of solvent or air-sensitive rea-
gents to the apparatus. Alternatively a balloon can be taped to a short piece
needle of thick-walled rubber pressure tubing to provide a supply of nitrogen when
locking nut stainless steel piston withdrawing air-sensitive reagents using a syringe (Fig. 19.6).
Teflon® tip
Syringe needles and cannulae
Fig. 19.4 Syringes for inert atmosphere work.
Stainless steel Luer lock syringe needles come in various lengths and diam-
eters. The length of needle you will need depends on the size of the vessel
from which you wish to withdraw the liquid; the diameter required depends
on the size of the syringe – you should not use a large-diameter needle with
a small-volume syringe – and the viscosity of the solution or liquid. Needle
diameters are expressed in ‘gauge’: the higher the gauge, the narrower the
Fig. 19.5 Needle-to-tubing connector. needle diameter. For most inert atmosphere work you should use a needle with
a ‘non-coring’ or ‘deflecting’ tip (Fig. 19.7), which ensures that a piece of the
balloon
Working with viscous liquids – septum is not trapped in the needle when you push it through. Cannulae are
attempting to draw the solution up a fine long, flexible, double-ended needles made from stainless steel or inert plastics,
tape from one vessel
which are used to transfer large volumes of reagents or solvents
needle creates a strong vacuum in the thick-walled rubber tubing
syringe, which may result in air being to another under inert gas pressure (Fig. 19.8). needle-to-tubing connector
pulled into the syringe via the small gap A generic method for transferring an air-sensitive reagent, by syringe,
Luer-lockfit® needle
between the syringe barrel and the pis- from a bottle to a pressure-equalising dropping funnel is described in Box 19.1.
ton. Use a larger diameter needle. (a)
balloon
syringe
balloon
ring support to hold
bottle securely
tape
reagent bottle
thick-walled rubber tubing
needle-to-tubing connector
Luer-lockfit® needle
stand
(a) (b)
Fig. 19.6 Inert atmosphere transfers: (a) balloon and needle; (b) preserving the
balloon
inert atmosphere while removing reagent.
syringe
ring support to hold

bottle securely
cannula
seal with inert gas out
septum
small septum
bubbler
inert gas
water out
inert gas in
clamp
ring clamp
clamp
water in
reaction vessel
magnetic ‘flea’
Fig. 19.7 Needle with non-coring tip for

Fig. 19.8 Transfer of air-sensitive reagents using the double-ended needle
piercing septa.
technique.
bubbler
seal with septum

syringe
septum
water out
inert gas in pressure-equalising
clamp addition funnel
condenser
clamp
water in
three-neck flask
‘flea’
stirrer
Fig. 19.9 Transferring an air-sensitive reagent to a pressure-equalising

dropping funnel.
Box 19.1 How to transfer an air-sensitive reagent using a syringe
1. Assemble the apparatus (Fig. 19.9) while hot and 3. Draw a few millilitres of solvent into the syringe, invert
allow to cool while nitrogen is flowing through it and it and squirt the solvent and any air into the waste
then add all the chemicals and solvents as required. solvent bottle. Repeat this step three times to ensure
that syringe and needle contain only solvent vapour.
2. Assemble the syringe and needle, making sure that
the needle is locked in place having first checked that 4. Using the syringe, inject a few millilitres of the dry
it is air-tight. solvent into the pressure-equalising dropping funnel,

either via the septum or by quickly removing the bottle. Make sure that you do not press the syringe
stopper while diverting the gas flow through funnel piston and squirt the reagent out of the syringe!
by putting your finger tip over the ‘bubbler’ outlet.
This ensures that there is an inert atmosphere in the 11. Inject the solution into the pressure-equalising drop-
funnel. Replace the stopper and release the gas flow ping funnel, either by:
through the ‘bubbler’.
(a) holding the syringe needle near the tip, piercing
5. Clamp the bottle of air-sensitive reagent to a support the septum on the pressure-equalising dropping
stand so that it cannot fall over while you manipulate funnel and injecting the reagent; or by
the syringe needles. (b) putting your finger over the ‘bubbler’ outlet
and removing the stopper from the pressure-
6. Remove the needle from the end of a needle-to- equalising dropping funnel. You can now inject
tubing connector carrying a balloon. Connect the
the solution into the funnel, protected by the nitro-
inert gas supply using clean, dry rubber tubing and
gen and solvent vapour coming out of the funnel
inflate the balloon. Turn off the gas supply, twist the
neck. Replace the stopper and release the gas flow
neck of the balloon to stop the gas escaping, reattach
through the ‘bubbler’.
the needle, ensuring it is locked in place, and release
the neck of the balloon. Dip the end of the needle into 12. Draw a few millilitres of dry solvent into the syringe
a little dry solvent to confirm that nitrogen is being and inject them into the pressure-equalising drop-
released from the needle – check for bubbles. ping funnel. This rinses any residual reagent from the
7. Hold the needle near the tip and pierce the septum syringe.
on the bottle. Make sure that the needle tip is in the 13. Draw some methanol or another solvent which reacts
space above the liquid. Support the needle connector gently with the reagent to destroy the air-sensitive
and balloon by a clamp on the support stand. reagent and squirt it into an excess of water. To clean
8. Holding the syringe and the needle near the tip, pierce the syringe assembly, draw water into the syringe
the septum on the bottle. Still holding the needle, ease several times and then disassemble it, wash well with
the syringe needle into the space above the liquid. propanone (acetone) and water, and then dry in the
oven.
9. Draw up some gas into the syringe and expel it back
into the bottle. Repeat three times and then ease the 14. Remove the needle–balloon assembly from the rea-
syringe needle into the liquid. gent bottle, cover the holes in the septum with a little
hydrocarbon grease and screw the bottle cap in place.
10. Draw up the required volume of solution into the Place the needle in the oven after washing with pro-
syringe and carefully ease the needle out of the panone (acetone) and then water.
Text references
Errington, R.J. (1997) Advanced Practical Inorganic and Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper-
Metalorganic Chemistry. Blackie Academic and Profes- imental Organic Chemistry, 2nd edn. Blackwell Science
sional, London. Ltd, Oxford.
Furniss, B.A., Hannaford, A.J., Smith, P.W.G. and Tatchell,
A.R. (1991) Vogel ’s Textbook of Practical Organic
Chemistry, 5th edn. Longman, Harlow, Essex.

Isac-Garcia, J., Dobado, J.A., Calvo-Flores, F.G. and Lehman, J.W. (2009) Student Lab Companion: Labo-
Martinez-Garcia, H. (2015) Experimental Organic ratory Techniques for Organic Chemistry, 2nd edn.,
Chemistry, Laboratory Manual. Elsevier, Amsterdam. Pearson, Harlow.

Keese, R., Brandle, M.P. and Toube, T.P. (2008) Practical Smart, L. (ed.) (2002) Separation, Purification and Iden-
Organic Synthesis: A Student’s Guide. John Wiley & tification – The Molecular World. Open University and
Sons Ltd, Chichester. Royal Society of Chemistry, Cambridge.
Leonard, J., Lygo, B. and Procter, G. (2013) Advanced Prac- Suib, S.L. and Tanaka, J. (1999) Experimental Meth-
tical Organic Chemistry, 3rd edn. CRC Press, Boca Raton. ods in Inorganic Chemistry. Prentice Hall, Harlow,
Loewenthal, H.J.E. and Zass, E. (1992) A Guide for the Essex.
Perplexed Organic Experimentalist, 2nd edn. John Zubrick, J.W. (2015) The Organic Chem Lab Survival
Wiley & Sons Ltd, Chichester. Manual. A student’s guide to techniques, 10th edn. John
Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical Wiley & Sons Ltd, Chichester.
Organic Chemistry. Chapman & Hall, London.
Study exercises
19.1 Look up and report on the information to be (a) ethoxyethane (diethyl ether)
found in the references and sources to dry the (b) tetrahydrofuran
solvents below to an appropriate level for inert (c) hexane.
atmosphere reactions:

20 Combinatorial chemistry
Traditionally practical chemistry has been focused on making pure samples of

target compounds, and the techniques have been devised to aid production of
individual samples of high purity. However, more recently it has been a priority
for synthetic chemists to develop methods which can make a large number of
structurally related compounds simultaneously.
The driving force for this change of emphasis has been a number of
areas such as medicinal chemistry where finding the most active compound
with the least deleterious effects can be best assessed by making a library
of as many slightly differing compounds to the lead compound as possible,
and then screening them for suitability. Often the importance of making as
many compounds as possible (in what is known as a ‘parallel synthesis’)
outweighs consideration of high purity, and often an acceptable level of
purity for library compounds made by combinatorial methods can be lower
than 95%, with the proviso that any library compound that has the appro-
priate properties will be remade at high purity, and testing redone on a high
purity sample.
There are many different methods used in what is quite a new branch of
synthetic chemistry, with no method having proven to be dominant, and devel-
oping new methods, reactions and equipment is an area of ongoing research
and development. We will look at two methods of creating a combinatorial
library – a solid phase combinatorial method and a parallel synthesis using a
multi-component reaction.
Solid phase combinatorial chemistry

Solid phase synthetic methodology has been developed from the pioneer-
ing work in the synthesis of proteins (polymers of amino acids) by Bruce
Merrifield. Instead of combining molecules and isolating and purifying at each
stage – resulting in losses and poor yields overall – Merrifield attached the first
molecule to an inert and insoluble polymer bead by a ‘linker’ and then carried
out chemical reactions to attach the second molecule. Since the new structure
was still attached to the insoluble polymer, no isolation or purification was
necessary. Repetition of the procedure allows the preparation of polymeric
structures with defined but complex organisation, e.g. biomolecules such as
insulin.
Solid phase synthesis lends itself to automation, since the reaction
product is always attached to the insoluble polymer bead, reagents and
reactants can be ‘washed’ past the polymer beads as can rinsing of the prod-
uct to free it from unreacted molecules. A computer-controlled machine can
complete a specific synthesis of a complex molecule by adding the correct
amounts of different chemicals and washing the products free of unused
reagents and finally detach the desired the final product from the polymer
beads. Variation in the order and type of molecules joined up leads to the
‘combinatorial library’.

Combinatorial chemistry
FG1(protected) FG1(protected)
FG1(protected) a b
P 1 P R1 FG2(protected) P R1 FG2
R1 FG2(protected)
FG1 FG3 FG1(protected) FG3(protected) d FG1(protected) FG3(protected)

e
P
R1 FG2 R2 FG4 R1 FG2 R2 FG4(protected) R1 FG2 R2 FG4(protected)
a Combine protected molecule with resin

b Deprotect one functional group
c Combine with new protected molecule
d Remove from polymer stationary phase
e Deprotect all functional groups to reveal new molecule
Fig. 20.1 A general schematic of a solid phase synthesis.
Solid phase synthesis is an extremely versatile process, allowing many var-

iations in chemical structure, and entails a lot of very clever organic chemistry:
●● different ‘linkers’ to attach different chemical classes to the polymer (amines,
phenols, alcohols, acids, etc.);
●● different protecting groups: these groups are attached to functional groups
(FGs) on the reacting molecules to prevent reaction during the synthetic
process, but can be removed (deprotection) as required at later stages without
affecting the overall synthesis;
●●removal of the completed molecule from the polymer, without affecting the
product molecule.
A general schematic of a section of a solid phase synthesis is shown in
Fig. 20.1.
Parallel synthesis using a multi-component reaction

An alternative parallel synthetic method is to use a multi-component reac-
tion where the diversity is created by varying the identity of the reagents in a
methodical way, and a microtitre plate (Fig. 20.2) enables many simultaneous
reactions to be carried out in a manageable way. Microtitre plates typically
have 96 wells (though they can be bought with more or fewer wells but always
laid out in a 2:3 rectangular matrix pattern), each of which is used to be the
reaction vessel for one set of reagents. For example, the Ugi reaction (Fig. 20.3)
can be used to make a bis-amide using four reagents: a ketone or aldehyde, an
amine, an isocyanide and a carboxylic acid. If there were 4 ketones, 4 amines,
3 isocyanides and 4 carboxylic acids available, then it is possible to make
96 different combinations of components and hence 96 different compounds.
Using a 96-well plate, it is possible to arrange the addition of reagents such
that each well yields a single component, and the plate itself contains a sample
of each of the possible 96 compounds. A further advantage of this method is
that many analytical techniques (such as UV/vis absorbance, fluorescence and
mass spectrometry) can be operated with robotic sample loaders which can
systematically analyse all wells on a microtitre plate.

Fig. 20.2 A microtitre plate
R2 O
2 1
R1 OH R2 C N R5
O R1 N R5
NH2 N
R3 R4
O R3 R4 O H
Fig. 20.3 The Ugi reaction.
Text reference
Merrifield, R.B. (1963) J. Amer. Chem. Soc., 85, 2149.

Anon. Combinatorial Chemistry-An Online Journal Bannwarth, W. and Felder, E. (eds) (2009) Combinatorial
Available: www.journals.elsevier.com/combinatorial- Chemistry: A Practical Approach, Volume 9. John Wiley
chemistry-an-online-journal Last accessed 06/01/16. & Sons Ltd., Chichester.
[A free, monthly literature review.]

Fassina, G. and Miertus, S. (eds) (2005) Combinatorial Jung, G. (ed.) (2008) Combinatorial Chemistry: Synthesis,
Chemistry and Technologies: Methods and Applications Analysis, Screening. John Wiley & Sons Ltd., Chichester.
2nd edn. CRC Press, Boca Raton.
Miller, B.L. (2010) Dynamic Combinatorial Chemistry: In
Fenniri, H. (ed.) (2000) Combinatorial Chemistry: A Prac- Drug Discovery, Bioorganic Chemistry and Materials
tical Approach. Oxford University Press, Oxford, UK. Science. John Wiley & Sons Ltd, Chichester.
Ganesan, A. (ed.) (2010) Combinatorial Chemistry: A Reek, J.N.H. and Otto, S. (eds) (2010) Dynamic Combina-
Primer. Elsevier Ltd, Amsterdam. torial Chemistry. John Wiley & Sons Ltd, Chichester.
Study exercises
20.1 If you were to make a library of tripeptides using a library of compounds. Assuming you have 12
a random combinatorial method with 20 amino chemicals which can act as A, 5 which can act as
acids, how many unique compounds would you B and 8 which can act as C, and that you wish to
expect to make? make all the possible variants, how many 96 well
20.2 You are investigating a multi-component reaction plates would you need to use?
(which uses three reagents, A, B and C) to make

Classical techniques
21. Qualitative techniques for inorganic analysis 179
22. Gravimetry 184
23. Molecular formulae 187
24. Procedures in volumetric analysis 190
25. Acid–base titrations 199
26. Complexometric titrations 203
27. Redox titrations 208
28. Precipitation titrations 211

21 Qualitative techniques for inorganic
analysis
Qualitative techniques are used to identify cations and anions in aqueous solu-
tion by simple reactions, mostly involving the production of a precipitate, evo-
lution of gas or a visual colour change. It is important to make observations
accurately and to interpret them in a step-wise manner.
The following basic equipment is required to carry out qualitative analysis:
●● Test tubes – in which the reactions are performed. (p. 65)
●● Sealing film or plastic stoppers – for the protection of the contents of test
tubes from contamination and for safe storage. (p. 67)
●● Test tube rack – this allows test tubes to be stored upright when not in use.
●● Test tube holder – this allows individual test tubes to be heated safely. (p. 177)
●● A glass rod – this has several functions including the stirring, transfer of
solutions, and the break-up of precipitates.
●● Watch-glasses these have several functions including the covering of beakers
to prevent contamination and as a receptacle for solutions.
●● A wash bottle containing distilled water.
Wash bottle – always keep a wash bot- ●● Spatula – for transferring small quantities of solids.
tle of distilled water handy.
●● Pasteur pipettes – for transferring liquids. (p. 63)
●● Micro-Bunsen burner – for heating solutions to boiling and for evaporating
Spatula – never place the spatula in solutions. (p. 172)
the test solution, it may lead to false-
positive tests for iron and chromium.
●● Evaporation crucible – this is used as a container for solutions when com-
plete evaporation of liquid is required, leaving a solid product.
●● Crucible tongs – for removing the crucible from the heat source.
●● Centrifuge – for separating precipitates from solution.
●● Heated water bath. (p. 173)
Reagents
Table 21.1 Typical reagents for qualitative At the start of your experimental work always check that the appropriate
analysis reagents are readily available (a list of commonly used reagents is given in
Table 21.1). Note that it is essential to use distilled water in all qualitative
2 M NH4OH Conc. HCl
analysis. Tap water contains ions such as Ca2+ , Mg 2+ , Fe2+ , Fe3+ , SO2-
4 and
2 M NaOH 0.1 M HNO3 Cl- and its use could lead to ‘false-positive’ results.
2 M AgNO3 2 M HNO3
2 M CH3COOH Conc. HNO3 Testing for anions and cations
2 M BaCl2 2 M H2SO4 Specific literature containing tests for the determination of anions and cations
0.1 M HCl Conc. H2SO4 can be found on p. 182. In general, however, the following tips are useful when
2 M HCl carrying out qualitative analysis:
●● Always work tidily to prevent cross-contamination of samples.
●● Ensure that all glassware has been cleaned thoroughly in detergent and then
rinsed twice with distilled water. Invert the test tubes to drain; never dry the
inner surface with towelling or tissue.

Qualitative techniques for inorganic analysis
Qualitative tests for cations and anions ●● Label test tubes at the start – it may prove difficult to remember what you
An unknown solution was tested as follows: have done later on.
Test Observation Conclusion ●● Always test solutions with a known composition before you attempt to
2 drops of dilute White Sulphate analyse solutions with an unknown content. This allows you to gain the
HCl, boil, then add 1 precipitate (SO2-
4 ) present necessary experience in solution manipulation, observation skills and the
drop BaCl2 solution
interpretation of results.
●● The colour of solutions and/or precipitates has to be interpreted from writ-
ten or oral information. Interpretation of colour can be subjective, so you
Conclusion will need to gain sufficient experience using solutions of known content to
Test performed on Report of the
unknown solution observations made drawn from the establish how a particular colour appears to you.
observation
●● Establish a protocol for recording of observations after carrying out different
Fig. 21.1 Recording your observations in tests (Fig. 21.1).
qualitative analysis.
●● Reagents should be added from Pasteur pipettes held with the tip just above
the mouth of the test tube. Never put Pasteur pipettes inside test tubes as this
can lead to contamination of the reagents.
●● Effective mixing of the test solution and added reagents is essential. This can
A clear solution is transparent – a
be achieved by holding the test tube between the thumb and index finger of
‘colourless’ solution has no colour.
one hand and ‘flicking’ the tube with the index finger of your other hand.
Alternatively, solutions can be mixed by bubbling air from a Pasteur pipette
held at the bottom of the test tube.
●● Evolved gas can be drawn up into a Pasteur pipette and then bubbled through
a test solution, e.g. CO2 can be drawn into a Pasteur pipette and then ‘blown’
out through lime water [Ca(OH)2 solution] giving a milky-white solution.
●● Solutions can be tested for pH using litmus paper. Never place litmus paper
directly into the test solution. Instead, dip a glass rod into the solution,
remove, touch the wet glass rod onto the litmus paper and note the colour.
Acidic solutions change blue litmus paper to red; alkaline solutions change
red litmus paper to blue. Alternatively, universal indicator paper can be
used. In this case, the orange paper turns ‘reddish’ with acidic solutions
and ‘bluish’ with alkaline solutions. By comparing any change in colour
with a chart (supplied with the universal indicator paper), the pH of the
solution can be estimated.
Centrifugation of solutions
Centrifugation causes particulate material to accumulate at the bottom of the
test tube. The procedure for centrifuging your sample is described in Box 21.1.
The speed and time of the run will depend on the centrifuge available, but will
typically be in the range 5000–10000 rpm for 5–10 min, respectively. Always
allow the centrifuge to stop in its own time, as abruptly halting the centrifuge
will disturb light precipitates. After centrifugation, hold the test tube at an angle
so that it is easy to remove the liquid component (or centrifugate) with a Pasteur
pipette (Fig. 21.2). You will find that it is difficult to remove all the centrifugate
residue
from the precipitate, and to maximise the transfer of centrifugate it is necessary
to wash the precipitate. This is carried out as follows:
●● Add a small quantity of distilled water to the precipitate.
liquid
●● Use a glass rod to break up the precipitate and mix thoroughly.
Fig. 21.2 Separation of liquid from a resi-
due using a pipette. ●● Recentrifuge the mixture.
180 Classical techniques

Box 21.1 How to use a low-speed bench centrifuge
1. Choose the appropriate test tube size, with stoppers 5. Load the sample test tubes into the centrifuge.
where necessary. Most low-speed machines have four- Make sure that the outside of the centrifuge tubes, the
place or six-place rotors. Use the correct number of sample holders and sample chambers are dry: any
samples to fill the rotor assembly whenever possible. liquid present will cause an imbalance during cen-
trifugation (as well as potentially causing corrosive
2. Fill the tubes to the appropriate level: do not over- damage to the rotor). Balanced tubes must be placed
fill, or the sample may spill during centrifugation. opposite each other; use a simple code if necessary,
3. Ensure that the rotor is balanced during use. To to prevent errors.
achieve this prepare identical test tubes and place 6. Bring the centrifuge up to operating speed by gentle
these diametrically opposite each other in the rotor acceleration. Do not exceed the maximum speed for
assembly. However, for low-speed work, where you the rotor and tubes used. If the centrifuge vibrates at
are using small amounts of particulate matter in any time during use, switch it off and find the source
aqueous solution it is sufficient to counter-balance a of the problem.
sample with a second test tube filled with water.
7. On completion of the run, allow the rotor to stop
4. If you are using centrifuges with swing-out rotors, spinning, release the lid, and remove all test tubes.
check that each holder/bucket is correctly posi-tioned in If any sample has spilled, make sure you clean it up
its locating slots on the rotor and that it is able to swing thoroughly.
freely. All buckets must be in position on a swing-out
rotor: even if they do not contain sample tubes, buck- 8. Finally, close the lid (to prevent the entry of dust)
ets are an integral part of the rotor assembly. and return all controls to zero.
●● Transfer the liquid obtained to the original centrifugate and store this solu-
Safety Note Take care when heat- tion for further analysis.
ing unknown solutions. As well as the
risk of burns, some reactions can be ●● Repeat the washing process, but this time discard the centrifugate, and retain
violent. the precipitate for further tests.
Heating test tubes and other containers

Safety Note Never point a test It is often necessary to heat a solution in a test tube, either to cause precipitation
tube towards yourself or, for that mat- or to dissolve a precipitate. You can carry out this heating effectively and safely
ter, towards anyone else while evapora- by partially immersing the test tube containing the mixture in a simmering
tion is being carried out. boiling-water bath (remember to use a test tube holder!).
It is possible to reduce the volume of the solution in the test tube, i.e.
to pre-concentrate the sample, by evaporation. Two different methods can be
Safety Note Never look down into employed.
a test tube (even with safety glasses
1. Transfer the solution to a small evaporating dish. Place the evaporating
on – there is still a risk of hot solution
dish on a wire gauze located on a tripod stand, and apply heat using a
suddenly being ejected into your face).
micro-Bunsen burner. Note that the volumes of solutions in qualitative
Always view coloured products through
analysis are often small, and excessive heating might result in hardening
the wall of the test tube.
of any residue, making it unusable.
2. Alternatively, evaporate the solution directly in a test tube by gentle heating
Safety Note Noxious fumes can over a micro-Bunsen burner. Remember to use a test tube holder. Position
be given off in some instances. Be care-
the test tube at an angle with the tip of the Bunsen burner flame positioned
ful not to breathe them in. Always work
at the upper surface of the liquid. Place a glass rod inside the test tube and
in a well-ventilated room or a fume
rotate constantly. This acts to disperse bubbles of steam that are given off.
cupboard.
Extreme caution is required with this method of evaporation, as the steam
bubbles can cause the solution to ‘bump’ out of the test tube (see p. 87).

‘Bumping’ can result in hot (and maybe toxic) substances being ejected
Beware – hot glass looks exactly like
over a surprisingly large distance.
cold glass.
Flame tests
Simple flame tests can be carried out on solid samples. Place a little of the solid
on a watch-glass and moisten with a drop of concentrated hydrochloric acid.
The purpose of the hydrochloric acid is to produce metal chlorides which are
volatile at the temperature of the Bunsen burner.
Pre-clean a platinum or nichrome wire by holding it in the hottest part of
the Bunsen flame (just above the central blue cone) until there is no coloured
flame from the wire. Cool, then dip the cleaned wire into the moistened solid
sample. Place the wire at the edge of the Bunsen flame (Fig. 21.3) and record
the colour of the flame from the sample (see Table 21.2).
Table 21.2 Flame tests for cations
Cation Flame colour
Barium Apple-green
Calcium Brick-red**
Copper Green
Potassium Lilac*
Sodium Intense yellow
Strontium Crimson**
Lead, arsenic, antimony and bismuth Dull blue
*The colour is often obliterated by trace impurities of

sodium present (sodium gives an intense yellow colour).
You can overcome this by viewing the colour through
cobalt-blue glass which allows the lilac coloration from
potassium to be seen.
**Viewing through cobalt-blue glass also allows calcium
and strontium to be distinguished. In this case, calcium is
Fig. 21.3 Holding a nichrome wire in a flame test. light green in colour while strontium appears purple.

Dean, J.A. and Patnaik, P. (2004) Dean’s Analytical Chem- Svehla, G. (1996) Vogel’s Qualitative Inorganic Analysis,
istry Handbook, 2nd edn. McGraw-Hill Co. Inc. 7th edn. Longman, Harlow, Essex.
Hardcastle, W.A. (1998) Qualitative Analysis. A guide to Svehla, G. (1989) Vogel’s Textbook of Macro and Semimi-
best practice. Royal Society of Chemistry, Cambridge. cro Qualitative Inorganic Analysis, 5th edn. Longman,
Kramer, B.K. and McCormick, J.M. (2013) Inorganic Harlow, Essex.
Qualitative Analysis. Available: http://www.chemlab Witten, KW., Davis R.E. Peck, M.L. and Stanley, G.G.
.truman.edu/CHEM131Labs/QualAnalysis.asp Last (2006) A Qualitative Analysis Supplement, 8th edn.
accessed 05/01/16. Thomson Brooks/Cole Publishing, Belmont, CA.
Lagowski, J.T. and Sorum, C.H. (2005), Introduction to
Semimicro Qualitative Analysis 8th edn. Prentice Hall,
Harlow, Essex.

Study exercises
21.1 Why is it necessary to use distilled water in qual- (b) A flame test gave a green flame.
itative inorganic analysis?
Deduce possible identities for the solid
21.2 Analysis of a solid gave the following analysis
compound.
results:
(a) Treatment of the solid with dilute HCl gave off

a gas, which when bubbled through a Ca(OH)2
solution gave a fine white precipitate.

22 Gravimetry
Gravimetric analysis is the process of converting an element into a definitive

compound, isolating this compound from other constituents in a sample and
then weighing the compound (Box 22.1). The weight of the element can then be
calculated from the formula of the compound and the relative atomic masses of
the elements involved. You need to be able to weigh accurately, by difference,
a substance to four decimal places (see p. 80).
Key Point The essential component of gravimetric analysis is

the transformation of the element of interest into a pure stable
solid compound, suitable for weighing.
The most common approach for isolating the element is by precipitation

from a solution where it is present in ionic form (see p. 106). Ideally, the con-
stituent under investigation is precipitated out of solution as a water-insoluble
compound, so that no losses occur when the precipitate is separated by filtra-
tion, washed free of soluble impurities and then weighed.
Box 22.1 How to carry out gravimetric analysis
Suppose that you wanted to analyse the amount of metal nickel dimethylglyoximate (Ni(HDMG)2) weighing
in an alloy. For example, you might want to determine the 0.2370 g.
nickel content, as a w/w percentage (p. 103), in a particu-
7. Write out the equation for the reaction and perform
lar sample of steel.
the calculation. In this example, you need to check
1. Select an appropriate solvent for your sample: in this the relative atomic masses of the elements involved.
example, you could dissolve the steel in aqua regia, a The relative molecular mass of nickel dimethylglyox-
combination of concentrated nitric acid and concen- imate is 288.91 g mol-1 (for structure see Fig. 22.1;
trated hydrochloric acid in the volume ratio of 1:3, Ar, H = 1.01; O = 16.00; C = 12.01; N = 14.01;
respectively. Ni = 58.69). The equation for the reaction can be
summarised as:
2. Choose an appropriate precipitant and carry out the
precipitation reaction. Here, an alcoholic solution of
Ni2+ + 2H2DMG = Ni(HDMG)2 + 2H +
dimethylglyoxime could be used to precipitate nickel
from a hot solution of aqua regia, by adding a slight According to the above equation, for each mole of Ni in
excess of aqueous ammonia solution, forming a red the steel sample, 1 mole of precipitate will be formed.
precipitate of nickel dimethylglyoximate (Fig. 22.1). Therefore, 0.2370 g of precipitate corresponds to:
3. Filter the precipitate through a pre-weighed Gooch
crucible. This should have been previously dried in an 0.2370 g Ni(HDMG)2 , 288.95 g mol-1 Ni(HDMG)2
oven at 120 °C and stored in a desiccator until required. = 8.20 * 10-4 mol Ni(HDMG)2
4. Wash the precipitate: in this example, the nickel The amount of nickel in the steel sample must there-
dimethylglyoximate can be washed with cold water fore be:
until qualitative testing shows that the wash solution
is free of chloride ions. 8.2 * 10-4 mol Ni(HDMG)2 * 58.69 g mol-1 Ni
5. Dry the precipitate in an oven, for example, at 120 = 0.0481 g Ni
°C, and allow to cool in a desiccator.
The percentage weight of nickel in the steel sample is
6. Determine the weight of the precipitated com- therefore:
pound. Suppose in this example that the nickel
had been precipitated from steel (2.0980 g) using 0.0481 g Ni , 2.0980 g sample * 100 = 2.29% w/w
dimethylglyoxime (H2DMG), giving a precipitate of

Gravimetry
O
H
O
Precipitation
CH3 C N N C CH3 Inorganic ions can be separated from mixtures using organic reagents (precip-
Ni
CH3 C N N C CH3 itants), with which they form sparingly soluble, often coloured, compounds.
O O The precipitants usually have high molecular weights, so a small quantity of the
H ion will produce a large amount of precipitate. Ideally, the precipitant should
Fig. 22.1 Structure of nickel– be specific for a particular ion, though this is rarely so. Examples of common
dimethylglyoxime complex. precipitants and their target ions are shown in Table 22.1 and their structures
are shown in Fig. 22.2.
Dimethylglyoxime is only slightly soluble in water (0.40 g L-1) and it is
Table 22.1 Common precipitants
therefore used as a 1% w/v solution in ethanol. Cupferron, the ammonium
Ion(s) of Possible salt of N-nitroso-N-phenylhydroxylamine, is used as a 5–10% w/v solution
Precipitant interest interferents in hydrochloric acid or sulphuric acid. Oxine (8-hydroxyquinoline) is almost
insoluble in water and is used as either a 2% or a 5% w/v solution in 2 mol L-1
Dimethylglyoxime Ni2+ Pd2+ , Pt2+ , Bi3+ acetic acid.
and Au3+
When precipitating a compound:
Cupferron Sn4+ Cu2+ and Pb2+
●● Mix your reagents slowly, with continuous stirring, to encourage the growth
8-Hydroxyquino- Al3+ Many metals
of large crystals of the compound.
line (oxine)
●● Improve the precipitation process by heating your solutions: ideally, one or
both solutions should be heated to just below boiling point.
CH3 C NOH
CH3 C NOH ●● Wash your precipitate with a dilute electrolyte solution, to remove any other
Dimethylglyoxime
constituents (it is essential to remove any impurities). Choose a solution
NO
that does not interact with the precipitate, and that is volatile at the drying
N temperature to be used.
O-NH+4
Cupferron ●● Use the minimum quantity of wash solution because no precipitate is abso-
lutely insoluble. While suitable wash solutions include dilute electrolytes,
N e.g. ammonium salts, ammonia solution or acids, pure water is rarely used,
OH as it may dissolve the precipitate.
8-Hydroxyquinoline
●● Test your filtered wash solution for impurities using simple qualitative tests
Fig. 22.2 Structures of common precipitants. (Chapter 21). Continue until your final washing solution contains no trace
of other constituents.
Filtering your precipitate – remember ●● It is best to wash repeatedly with several small amounts of solution, allowing
that the particle size of your precipitate the precipitate to drain between washings.
must be such that it is not lost during
the filtering process. Filtration
To carry out this procedure, you will need to assemble a Gooch crucible
Gooch crucible and funnel on a Büchner flask, clamped for stability using a retort stand
rubber cone (Fig. 22.3). The glass Gooch crucible has a porous disk of sintered ground
Gooch funnel glass, typically of pore diameter 20930 mm, which is satisfactory for moder-
ately sized precipitates. Fit the crucible into a Gooch funnel using a rubber
cone, put the funnel into a one-holed rubber bung and then into a Büchner
flask. The tip of the funnel must project below the side arm of the flask to
Büchner flask to pump
prevent loss of filtrate down the side arm (see p. 84). Then, connect the Büch-
ner flask to a water pump. Pour your precipitate suspension into the Gooch
crucible, using a glass rod (p. 75) to direct the liquid into the centre of the sin-
tered base. The lower end of the glass rod should be close to, but not touching,
the sintered-glass base. Never overfill the crucible. The precipitate remaining
in the bottom of the beaker should be rinsed out with the filtrate solution:
Fig. 22.3 Experimental arrangement for fil- disconnect the pump and pour the filtrate back into the beaker containing the
tration of precipitate.

Gravimetry
precipitate. The pump should be disconnected by pulling off the vacuum tube
from the Büchner flask. On no account turn off the water pump while doing
this (p. 86). You may need to rinse the beaker several times, to collect all of
the precipitate in the crucible.

Harris, D.C. (2010) Quantitative Chemical Analysis, 8th Rubinson, J.F. and Rubinson, K.A. (2003) Contemporary
edn. W.H. Freeman Co., New York. Chemical Analysis, 2nd edn. Pearson, Harlow, Essex.
Mendham, J., Denney, R.C., Barnes, J.D. and Thomas, Skoog, D.A., West, D.M., Holler, F.J. and Crouch, S.R.
M.J.K. (2000) Vogel’s Textbook of Quantitative Chemi- (2014) Fundamentals of Analytical Chemistry, 9th edn.
cal Analysis, 6th edn. Prentice Hall, Harlow, Essex. Brooks Cole, Belmont, CA.
Study exercises
22.1 Calculate the salinity of sea water (simply completeness of precipitation. The suspension
expressed as a % (w/w) of NaCl in water) from was allowed to cool in the dark (30 min) and then
the following experimental data: sea water filtered through a pre-weighed sintered crucible
(10.00 mL) was transferred to a tall beaker, and (porosity 4) using dilute solution of nitric acid [3
concentrated nitric acid (0.5 mL) and deionised mL of 2 M dissolved in deionised water (300 mL)]
water (200 mL) added. Silver nitrate solution for transfer and washing. The precipitate and cru-
(0.1 M) was added drop-wise from a burette cible were dried to constant weight to yield silver
with constant stirring until coagulation of the chloride (0.8329 g).
precipitate began to occur. The suspension was
22.2 Why is deionised water used in the dilution and
heated to boiling and stirred (5 min) and a few
washing processes in 22.1?
drops of silver nitrate solution added to confirm

23 Molecular formulae
When you have synthesised a new molecule/compound you must confirm its
structure. This can be achieved by identifying its molecular formula (MF) and
then its structure. Molecular formulae can be established by elemental microa-
nalysis or high resolution mass spectrometry (Chapter 38) and the structure is
deduced via infrared (Chapter 36), 1H@ and 13C@NMR (Chapter 37) spectros-
copy and mass spectrometry (Chapter 38).
Once you have purified your new molecule by recrystallisation (Chapter 14)
or distillation (Chapter 16) or preparative chromatography (Chapter 32)
or a combination of techniques and established its purity by melting point
(Chapter 13) and the appropriate analytical chromatographic and/or spec-
troscopic procedures, it is usual to submit a sample of the compound (about
200 mg) to a specialist analytical organisation to provide the data for determi-
nation of the molecular formula.
Elemental microanalysis
For the non-metallic elements, C, H, and N, the sample is combusted at high
temperature in oxygen and C and H are converted into CO2 and H2O, respec-
tively and N into nitrogen and nitrogen oxides which are further reduced to
N2. All three gases are then determined by thermal conductivity. Halogens
and sulphur are thermally decomposed in oxygen and estimated by micro-
volumetric techniques (Chapter 28) and metals can be estimated using ICP-
AES (Inductively Coupled Plasma-Atomic Emission Spectroscopy, p. 234) or
other classical analytical techniques. Recent developments enable oxygen to
be estimated as CO2.
The results of microanalysis are presented as % composition, e.g. for
propanamide
C, 49.28; H, 9.66; N, 21.89 %[23.1]
Key Point Unless specifically requested, the oxygen content is

NOT given and you must subtract the sum of the elemental per-
centages from 100 to obtain a value. ALWAYS check that the total
of the analytical data is 100; if not subtract the total from 100.
The complete analysis data for propanamide becomes

Definition
C, 49.28; H, 9.66; N, 19.16; O, 21.89 %[23.2]
Empirical formula (EF): the smallest num-
ber ratio of the elements in a compound The method of calculating the Empirical Formula (EF) and Molecular
obtained from microanalytical data. The Formula (MF) of a compound using the relative molecular mass (Mr) obtained
molecular formula (MF) is a multiple of from the low resolution mass spectrum (Chapter 38 is illustrated in Box 23.1.
the EF corresponding to the Mr. The procedure is applicable to inorganic and inorganic complexes (p. 203) as
well as organic molecules as exemplified in the following,
Examples:
Ethyne MF = C2H2, EF = CH CuSO4.5H2O Cu, 25.47; H, 4.00; S, 12.82; O, 57.7 %
Benzene MF = C6H6, EF = CH
Methanal MF = CH2O, EF = CH2O Co(H2NCH2CH2NH2)3Cl3 Co, 17.05; C, 20.85, H, 6.95; N, 24.33; Cl, 30.82 %
Glucose MF = C6H12O6, EF = CH2O but has less applicability in systems such as polymers and large biomolecules.

Molecular formulae
Box 23.1 How to calculate a molecular formula from microanalytical data
Example 1: Using the data from ’propanamide’ in [23.1]. Example 2: Compound X gave the following microanalyt-
ical data C, 58.87; H, 9.88% and Mr = 102
1. Check that the components total 100% as in [23.2]
2. Divide each value by the respective Ar 1. Check that the components total 100%.They do not, so
Compound X contains oxygen. 100 - 58.87 - 9.88 =
49.28 9.66 19.16 21.89 31.37 and required data is C, 58.87; H, 9.88; 0,31.37
C = H = N = O =
12.01 1.008 14.01 16.00 2. Divide each value by the respective Ar
become 58.87 9..88 31.37

C = H = O =
12.01 1.008 16.00
C = 4.073 H = 9.58 N = 1.359 O = 1.368 become
3. Divide each value by the smallest C = 4.90 H = 9.80 O = 1.96
4.073 9.58 1.359 1.368 3. Divide each value by the smallest

C = H = N = O =
1.368 1.368 1.368 1.368 4.90 9.80 1.96
C = H = O =
1.96 1.96 1.96
become
become
C = 2.977 H = 7.003 N = 0.993 O = 1.00
C = 2.50 H = 5.00 O = 1.00
4. Rounding to the nearest whole numbers gives the 4. This gives the Empirical formula (EF) as C2.5H5o but
Empirical formula (EF) as C3H7no you cannot have half or even a third of an atom.
5. Mr from the low resolution mass spectrum is 73 and 5. Mr from the low resolution is 102 and therefore the
therefore the Empirical formula (EF) is identical to the Empirical formula (EF) is half of the Molecular for-
Molecular formula (MF) and ’propanamide’ has the mula (MF) and Compound X has the molecular
molecular formula C3H7no f ormula C5H10o2
Whenever a new compound is synthesised it is normal for the theoretical

elemental analysis values to be submitted for the analysis process. This aids the
analysers’ calibration of their techniques (the appropriate sensitivity range can
be selected) and a good comparison between the theoretical and experimental
values is essential for publication.
High resolution mass spectra (HRMS)
Mr MF
Mr data from low resolution mass spectra (LRMS) are based on the sums of
the most abundant isotope masses in a molecule and result in whole number
73.0164 C2H3NO2 Mr values as shown in Chapter 38. High resolution mass spectra (HRMS) takes
73.0402 C2H5N2O into account that Ar values are not whole numbers and the contributions of
73.0641
isotopes to the overall Mr must be taken into consideration. The Mr derived
C2H7N3
from HRMS is accurate to at least four decimal places and a unique molecular
73.0289 C3H5O2 formula (MF) can be obtained from tables or data bases. Fig. 23.1 illustrates
73.0528 C3H7no the eight possible MF’s for a LRMS value of 73. Further spectroscopic analysis
73.0767
leads to confirmation of the molecular structure.
C3H9N2
73.0653 C4H9O Double bond equivalents (DBE)
73.0892 C4H11N
A further piece of information about the structure of a molecule can be obtained
Fig. 23.1 HRMS data for LRMS Mr = 73 from the MF. A simple calculation using eqn [23.3] gives rise to the double
indicating a possible MF bond equivalent (DBE).

Molecular formulae
[(2a + 2) - (b - d + e)]
For a molecule CaHbOcNdHale DBE = [23.3]
2
DBE = 1 implies one double bond or a saturated ring; DBE = 2 implies
two double bonds or one triple bond or saturated rings or an unsaturated ring.
Higher DBE values indicate a combination of these possibilities as the follow-
ing examples demonstrate:
6 + 2 - (6 - 0 + 0)
Example 1 propanone C3H6O DBE =
2
= 1 (one C “ O)
12 + 2 - (10 - 0 + 0)
Example 2 cyclohexanone C6H10O DBE =
2
= 2 (one C “ O, one ring)
6 + 2 - (7 - 1 + 0)
Example 3 propanamide C3H7ON DBE =
2
= 1 (one C “ O)
14 + 2 - (6 - 0 + 0)
Example 4 benzoic acid C7H6O2 DBE =
2
= 5 (one C “ O,
three C “ C, one ring).
The actual nature of the unsaturation can be ascertained by the spectroscopic

techniques described in Chapters 36, 37 and 38 and appropriate chemical tests.

Anon. Molecular Formula from HRMS. Available: http:// Silverstein, R.M., Webster, F.X., Kiemle, D.J. and
chemcalc.org/mf Bryce, D.L. (2015) Spectrometry Identification of
Last accessed 28/01/2016. Organic Compounds, 8th edn. John Wiley & Sons Ltd,
[Calculator of possible MF from HRMS data.] New York.
Crews, P., Rodriguez, J. and Jaspars, M. (2010) Organic
Structure Analysis, 2nd edn. Oxford University Press,
New York.
Study exercises
23.1 Calculate the EF and MF from the following data 23.2 Using eqn [23.3] calculate the DBE for the
using the Ar values: C = 12.01, H = 1.008, following:
n = 14.01, o = 16.00, Cl = 35.5.
(a) C3 H 5 N
(a) C, 54.86; H, 13.51; O, 21.62 % LRMS Mr = 74 (b) C8H14O2
(b) C, 53.33; H, 11.11 % LRMS Mr = 90 (c) C5H11N
(c) C, 54.55; H, 9.09 % LRMS Mr = 88 (d) C10H8
(d) C, 56.25; H. 3.90; Cl, 27.34 % LRMS Mr = 128

24 Procedures in volumetric analysis
Volumetric analysis, also known as titrimetric analysis, is a quantitative tech-

nique used to determine the amount of a particular substance in a solution of
unknown composition.
This requires:
●● A standard solution, which is a solution of a compound of accurately known
concentration, that reacts with the substance to be analysed.
Definition ●● The test solution, containing an unknown concentration of the substance to
be analysed.
A stoichiometric titration is one with
a known reaction path, for which a ●● Some means of detecting the end-point of the reaction between the standard
chemical reaction can be written, and test solutions, e.g. a chemical indicator or, in the case of potentiometric
and having no alternative or side titrations, a pH electrode (see Chapter 34). Some reactions exhibit a colour
reactions. change at the end-point without the addition of an indicator.
The volume of standard solution that reacts with the substance in the test solu-
tion is accurately measured. This volume, together with a knowledge of con-
centration of the standard solution and the stoichiometric relationship between
the reactants, is used to calculate the amount of substance present in the test
solution. Specific examples of the different types of calculations involved are
shown in Chapters 25–28.
Classification of reactions in volumetric analysis

There are four main types of reaction
1. Acid–base or neutralisation reactions, where free bases are reacted with a
standard acid (or vice versa). These reactions involve the combination of
hydrogen and hydroxide ions to form water.
2. Complex formation reactions, in which the reactants are combined to form a
soluble ion or compound. The most important reagent for formation of such
complexes is ethylenediamine tetra-acetic acid, EDTA (as the disodium salt).
3. Precipitation reactions, involving the combination of reactants to form a
precipitate.
4. Oxidation–reduction reactions, i.e. reactions involving a gain (reduction)
or loss (oxidation) of electrons. The standard solutions used here are either
oxidising agents (e.g. potassium permanganate) or reducing agents (e.g.
iron (II) compounds).
What can be measured by titration?
●● The concentration of an unknown substance, e.g. 0.900 mol L-1.

●● Percentage purity, e.g. 56%.
●● Water of crystallisation, e.g. (NH4)2SO4.nH2O.
●● Percentage of a metal in a salt, e.g. 12% Fe in a salt.
●● Water hardness, e.g. determination of the concentration of calcium and mag-
nesium ions.
Examples of the types of calculations used in volumetric analysis are shown
in Box 24.1.

Procedures in volumetric analysis
Box 24.1 Types of calculations used in volumetric analysis – titrations
In titrations you react a solution of a known concentration with a solution of an unknown concentration.
If you know the mole ratio of the two reacting chemicals in solution, you can calculate the amount (the number of
moles and thus the number of grams) of the solute in the solution of unknown concentration.
Let’s look at the reaction between NaOH and HCl:
HCl + NaOH = NaCl + H2O

Since the equation is balanced we know that 1 mol (36.5 g) of HCl will react with 1 mol (40 g) of NaOH. We know that
a 1.0 M solution of HCl contains 1 mol of HCl in 1000 mL (1 litre) of water. Then:
1000 mL of 1.0 M HCl solution is equivalent to 1.0 mol of NaOH

is equivalent to 1000 mL of 1.0 M NaOH solution
is equivalent to 40 g of NaOH
is equivalent to 23 g of Na+ ions
is equivalent to 17 g of OH - ions
Similarly, for the reaction between potassium hydroxide and sulphuric acid:
H2SO4 + 2KOH = K2SO4 + 2H2O
Since 1 mol of H2SO4 reacts with 2 mol of KOH, then:
1000 mL of 1.0 M H2SO4 solution is equivalent to 2.0 mol of KOH

is equivalent to 2 * 1000 mL of 1.0 M KOH solution
is equivalent to 2 * 56 g of KOH
is equivalent to 2 * 39 g of K + ions
is equivalent to 2 * 17 g of OH - ions
To work out the results of titrations you must always:
●● Work out the balanced equation to find out the ratio of moles reacting.
●● Decide what you are trying to calculate.
Example: 25.00 mL of sodium hydroxide solution were titrated by 24.00 mL of 0.1 M HCl solution. Calculate the con-
centration of the sodium hydroxide solution.
●● HCl + NaOH = NaCl + H2O
●● Concentration of NaOH, i.e. moles of NaOH in 1000 mL, since concentration is mol L-1.
Now:
1000 mL of 1.0 M HCl solution is equivalent to 1.0 mol of NaOH
but the concentration of HCl is only 0.1 M:
1000 mL of 0.1 M HCl solution is equivalent to 0.1 * 1.0 mol of NaOH
but only 24.00 mL of HCl solution were used:
1.0 * 0.1 * 1.0

1.0 mL of 0.1 M HCl solution is equivalent to mol of NaOH
1000
and
24 * 1.0 * 0.1 * 1.0
24 mL of 0.1 M HCl solution is equivalent to mol of NaOH
1000
(continued)

= 2.4 * 10-3 mol of NaOH

but 25.00 mL of NaOH solution were used:
25.00 mL of NaOH solution contains 2.4 * 10-3 mol of NaOH
Then
2.4 * 10-3
1.0 mL of NaOH solution contains mol of NaOH
25
and
1000 * 2.4 * 10-3

1000 mL of NaOH solution contains mol of NaOH
25
= 0.096 mol of NaOH
Therefore concentration of NaOH solution is 0.096 mol L-1.

Using this set of equations you can calculate directly the mass of NaOH per litre, the mass or moles of sodium ions
and the mass or moles of hydroxide ions.
Note: The expression [C1]V1 = [C2]V2 was not used, even though it is applicable in this case.
Problems arise when [C1]V1 = [C2]V2 is used for reactions which are not 1:1, e.g.:
H2SO4 + 2KOH = K2SO4 + 2H2O
or
2MnO4- + 16H + + 5C2O2-

4 = 2Mn
2+
+ 10CO2 + 8H2O
Titrations
The process of adding the standard solution to the test solution is called a
Definition
titration, and is carried out using a burette (see below). The point at which the
A primary standard should be easy reaction between the standard solution and the test substance is just complete
to obtain in a pure form. It should be is called the equivalence point or the theoretical (or stoichiometric) end-point.
unaffected in air during weighing, be This is normally detected by a visible change, either of the standard solution
capable of being tested for impurities itself or, more commonly, by the addition of an indicator.
and be readily soluble under the Standard solutions
conditions used. Finally, the reaction
A standard solution can be prepared by weighing out the appropriate amount of
with the standard solution should be
a pure reagent and making up the solution to a particular volume, as described
stoichiometric and instantaneous.
on p. 72. The concentration of a standard solution is expressed in terms of
molarity (p. 102). A substance used in a primary standard should fulfil the
following criteria:
●● It should be obtainable in high purity (7 99.9%).
●● It should remain unaltered in air during weighing (i.e. it should not be
hygroscopic).
●● It must not decompose when dried by heating or vacuum.
●● It should be capable of being tested for impurities.

●● It should be readily soluble in an appropriate solvent.

●● It must react with the test substance stoichiometrically and rapidly.
Preparing a standard solution

The molarity of a solution is the concentration of the solution expressed as
mol L-1. If xg of a substance of molecular weight Mr is dissolved in ymL of
x
distilled water, the moles of substance dissolved = = m.
Mr
m * 1000
Therefore, molarity (mol L-1) = [24.1]
y
Primary standards prepared from solid compounds should be weighed out
using the ‘weighing by difference’ method as described in Chapter 9, and accu-
rately made up to volume using a volumetric flask. Complete transfer of the
substance from the weighing vessel to the volumetric flask is best achieved by
inserting a funnel into the neck of the flask (Fig. 24.1). As much of the solid
as possible should be transferred via the funnel. The funnel should be washed
with distilled water prior to removal. Distilled water is then added to the flask,
with occasional swirling to help to dissolve the solid. This is continued until
the meniscus is about 1 cm below the volume mark. At this point a stopper is
inserted and the flask is inverted several times to ensure the solid is completely
dissolved. Finally, using a Pasteur pipette, distilled water is added up to the
volume mark. The solution should be thoroughly mixed before use.
If the solid is not readily soluble in cold water it may be possible to dis-
solve it by stirring in warm water in a beaker. After allowing the solution to
cool to room temperature, it can be transferred to the volumetric flask using a
glass rod and filter funnel (Fig. 24.1) followed by several rinses of the glass rod/
filter funnel. Finally, the solution is made up to the mark with distilled water.
For accurate readings – always remem-
ber to position your burette vertically.
Filling a burette
Titrand – this is the solution of
unknown composition in the conical ●● Clamp a clean 50.00 mL burette (p. 64) in a laboratory stand. Place a beaker
flask. The titrant is the standard solution on a white tile immediately below the outlet of the burette (Fig. 24.2).
in the burette. ●● Place a small filter funnel on top of the burette and, with the tap open, carefully
pour in the standard solution (or titrant) until it starts to drain into the beaker.
Safety Note Never mouth pipette. ●● Close the burette tap, and fill the burette with the standard solution until the
meniscus is about 1–2 cm above the zero mark. Remove the funnel.
Contamination of pipette fillers – this

●● Open the tap and allow the solution to drain until the meniscus falls to the
can affect your results detrimentally.
zero mark. The burette is then ready for the titration.
The main cause is sucking air into the Note that, to avoid contamination, the solution in the beaker should be dis-
pipette while filling and the bubbles of carded, rather than recycled.
liquid shoot into the pipette filler.
Using a pipette
Don’t lift the pipette out of the liquid
while filling. A clean 25.00 mL pipette (p. 64) is commonly required together with a suitable
pipette filler. Various designs of pipette filler are available. The most common
Ensure that there is more liquid in the type is based on a rubber-bulb suction device. It is best to evaluate a range of
beaker than the volume of the pipette. pipette fillers, if available in the laboratory, for ease of use and performance.
The pipetting procedure is given in Box 24.2.

glass rod
solid burette
clamp
retort stand
rinse
solution
Fig. 24.1 Quantitative transfer of a solid to a vol-

umetric flask.
Fig. 24.2 Apparatus for a titration.
Performing a titration
There are many minor variations in technique in performing titrations depend-
ent upon the type of titration (acid–base, redox, precipitation or back-titration)
being carried out. A robust basic procedure which can be applied to the majority
of analyses is given in Box 24.3.

Box 24.2 How to fill a pipette
1. Attach the pipette filler to the pipette as shown in pipette to eye level and gently squeeze valve ’E’,
Fig. 8.5. which allows the titrand to drain slowly back into the
beaker. Continue until the bottom of the meniscus is
2. Pour the solution of unknown composition (the titrand)
on the graduation mark.
into a clean dry beaker making sure that the amount
of solution well exceeds the volume of the pipette. 5. Wipe the outside of the pipette with a tissue
Never place the pipette in the volumetric flask contain- (Fig. 24.3(b)). Be careful not to touch the point of
ing the solution as this can lead to contamination of the the pipette with the tissue otherwise solution will be
solution from the external surface of the pipette. lost through capillary action.
3. Squeeze the valve ‘A’ on the pipette filler and then 6. Squeeze valve ‘E’ to allow the pipette’s contents to
squeeze the bulb to create some ’suction’. Dip the tip drain into a conical flask.
of the pipette into the solution and squeeze the valve
‘S’. Draw up the titrand until it is well above the gradu- 7. Still squeezing valve ‘E’, touch the tip of the pipette
ation mark (Fig. 24.3(a)). Squeeze valve ’E’ to empty the on the inside of the conical flask to ensure that the
pipette to waste – this rinsing process ensures that the correct volume is transferred (Fig. 24.3(c)).
titrand used subsequently will be undiluted and uncon-
Note: it is normal for a small quantity of solution to remain
taminated by any residue or liquids in the pipette.
in the pipette tip. This volume is taken into account when
4. Refill the pipette until the meniscus of the titrand pipettes are calibrated, so DO NOT attempt to ’blow out’
is above the graduation mark. Carefully raise the the liquid into the conical flask.
Box 24.3 How to carry out a titration

Note: this procedure is applicable to a right-handed increases and therefore the amount of titrant added
person. must be reduced.
1. Add one or two drops of indicator to the titrand in a 5. Very close to the end-point you will be adding the
conical flask. titrant at one drop at a time.
2. Hold the conical flask in your right hand and con- 6. When the new indicator colour ’flashes’ through the
trol the tap on the burette with your left hand. The solution, you are probably one drop of titrand away
burette should be arranged so that the tap is on the from the end-point. Note the volume used (Fig. 24.5)
opposite side of the burette to your palm. In this way, and then continue the titration by drops until the
your left hand also supports the body of the burette colour change of the indicator is permanent. Do not
(Fig. 24.4). forget to count the number of drops (see ‘Enhancing
precision’ below).
3. Add some titrant by opening the tap. Where the
titrant meets the titrand solution a colour change of 7. Refill the burette to the zero mark using a funnel,
the indicator will be observed. Swirl to titrand solu- ready for the next titration.
tion and the original colour will be restored.
8. Repeat the titration until consistent results (within
4. Continue adding titrant, but observe that as the 0.2 mL) are obtained.
titration proceeds to the end-point the length of
9. Do not average analytical results with large variations
time for the original indicator colour to be restored
(>0.2 mL).
Enhancing precision of a titration

You can increase the precision of your titrations by titrating ‘to the nearest drop’.
This means that you can measure end-points in a titration to a very small volume of
titrant, i.e. one drop. Before you commence the titration set the tap on the burette
to deliver a steady series of drops and count the number of drops in, say, 1.00 mL;
you can then calculate the volume of a drop. After carrying out the first ‘rough’

evacuate
bulb
fill
Reading a burette – your eye-line pipette
should be level with the bottom of the
meniscus. Then, record the volume
used to one decimal place.
empty
pipette
Rough titration – always carry out an

initial rough titration to determine the
approximate volume of titrant required
for the end-point. This allows you to
anticipate the end-point in subsequent
titrations to determine the accurate
volume.
(a) (b) (c)
Fig. 24.3 Using a pipette.
For consistency of results – always

perform two or three titrations (or until
consistent results are obtained, i.e. titre
values within 0.1–0. 5 mL).
Fig. 24.5 Reading a burette. Place

a white card below the level of the
meniscus. This allows an accurate
Fig. 24.4 Performing a titration. reading to be made.
Washing the test solution – any dis-

titration you can stop the addition of titrant, say, x mL before the end-point and
tilled water used for washing the test
add the titrant by drops until the end-point is reached. The volume of titrant
solution from the walls of the conical
used is then x mL + y drops, and since you have measured the volume of 1
flask has no effect on the titration or the
drop you now have a titration ‘to the nearest drop’. With practice you can titrate
calculation.
‘to half a drop’ by allowing half a drop to form on the burette tip and washing
it into the titrand solution with distilled water.

Volumetric analysis – calculating the concentration of a

test substance
The calculation should be carried out in a logical order as follows:
1. Write the balanced equation for the reaction between the standard and the
test substance.
2. From the stoichiometry of the reaction, determine how many moles of the
test substance react with 1 mole of the standard substance. For example,
in the reaction between an H2SO4 standard solution and an NaOH test
solution:
H2SO4 + 2NaOH S Na 2SO4 + 2H2O[24.2]
Therefore 2 moles of NaOH react with 1 mole of H2SO4.

3. Calculate the number of moles of standard substance used to reach the
end-point of the reaction. This can be determined from knowledge of the
concentration of the standard solution (mol L-1) and the volume of titrant
used (mL). Remember to take care with units – in this instance division
by a factor of 1000 is required to convert mL to L:
concentration (mol L-1) * volume (mL)

Number of moles =
1000
4. The number of moles of test substance present in the titrand is then obtained
from knowledge of the equivalences. In the example given above (point 2),
the number of moles of test substance is twice the number of moles of
standard substance. Therefore, if X moles of H2SO4 are used (as calculated
in point 3), 2X moles of NaOH were present in the initial volume of test
solution.
5. Finally, the concentration of the test solution can be calculated using the
formula:
Concentration of test 1000 * amount of test substance (mol)

=
solution (mol L-1) initial volume of test solution (mL)
Again, the factor of 1000 is used to convert mL to L.

Christian, G.D. Dasgupta, P.K. and Schug, K.A. (2014) Chemical Analysis, 6th edn. Prentice Hall, Harlow,
Harris, D.C. (2010) Quantitative Chemical Analysis, 8th Cambridge.
edn. W.H. Freeman Co., New York. Rubinson, J.F. and Rubinson, K.A. (1998) Contemporary
Jander, G., Jahr, K.-F., Schulze, G. and Simon, J. (2009) Volu- Chemical Analysis. Prentice Hall, Harlow, Essex.
metric Analysis, 17th edn. Walter de Gruyter Inc., Germany. Skoog, D.A., West, D.M., Holler, F.J., and Crouch, S.R.
Mendham, J., Denney, R.C., Barnes, J.D. and Thomas, (2014) Fundamentals of Analytical Chemistry, 9th edn.
M.J.K. (2000) Vogel ’s Textbook of Quantitative Brooks Cole, Belmont, CA.

Study exercises
24.1 Apply the methodology from Box 24.1 to the (ii) Sodium hydroxide solution (25.00 mL) was
following: titrated by sulphuric acid solution (24.0 mL;
0.05 M). Calculate the concentration of the
(i) Sodium carbonate solution (25.00 mL) was
sodium hydroxide solution.
titrated by sulphuric acid solution (26.0 mL;
(iii) Your burette reading for a titration was 25.20
0.1 M). Calculate the concentration of the
mL plus 8 drops. You have measured that
sodium carbonate solution.
12 drops = 1.00 mL. What is the total volume
of titrant used?

25 Acid–base titrations
The titration of an acid solution with a standard solution of alkali will determine
the amount of alkali which is equivalent to the amount of acid present (or vice
versa). The point at which this occurs is called the equivalence point or end-
point. For example, the titration of hydrochloric acid with sodium hydroxide
can be expressed as follows:
NaOH(aq) + HCl(aq) S NaCl(aq) + H2O(l)[25.1]

Safety Note Never pipette by If both the acid and alkali are strong electrolytes, the resultant solution will be
mouth. neutral (pH7). If on the other hand either the acid or alkali is a weak electrolyte
the resultant solution will be slightly alkaline or acidic, respectively. In either
case, detection of the end-point requires accurate measurement of pH. This can
be achieved either by using an indicator dye, or by measuring the pH with a
thymol blue glass electrode (described in Chapter 12).
H3C CH3 H3C CH3 Acid–base indicators

CH CH
H3C OH H3C OH Typical acid–base indicators are organic dyes that change colour at or near the
HO+ O
equivalence or end-point. They have the following characteristics:
H H
C C CH3 C C CH3
CH3
SO-3
CH3
SO-3
●● They show pH-dependent colour changes.
CH3 CH3
●● The colour change occurs within a fairly narrow pH range (approximately
red yellow 2 pH units).
(<pH 1.7) (pH 1.7 – 8.9)
●● The pH at which a colour change occurs varies from one indicator to another,
and it is possible to select an indicator which exhibits a distinct colour change
phenolphthalein
at a pH close to the equivalence or end-point.
HO OH O- O Selected common indicators together with their pH ranges and colour changes
are shown in Table 25.1. Examples for thymol blue and phenolphthalein are
O O-
C O
shown in Fig. 25.1.
C
O
Table 25.1 Colour changes and pH range of selected indicators

colourless red
(<pH 6) (pH 8.3 – 10)
Colour in acid Colour in alkaline
Fig. 25.1 Examples of indicators used Indicator pH range solution solution
in acid–base titrations: thymol blue and
Thymol blue 1.2–6.8 Red Yellow
phenolphthalein.
Methyl orange 2.8–4.0 Red Yellow
Methyl red 4.3–6.1 Red Yellow
Phenol red 6.8–8.2 Yellow Red
15
Phenolphthalein 8.3–10.0 Colourless Pink/red
10
Neutralisation curves
pH
5 A plot of pH against the volume of alkali added (mL) is known as a neutralisa-

tion or titration curve (Fig. 25.2). The curve is generated by a ‘potentiometric
0 titration’ in which pH is measured after each addition of alkali (or acid). The
0 50 100 150 200
significant feature of the curve is the very sharp and sudden change in pH near
volume of NaOH added (mL)
to the equivalence point of the titration. For a strong acid and alkali this will
Fig. 25.2 A typical neutralisation curve: occur at pH 7. If either the acid or base concentration is unknown, a prelimi-
0.1 M HCl with 0.1 M NaOH. nary ‘rough’ titration is necessary to find the approximate equivalence point

Acid–base titrations
14 followed by a more accurate titration as described on p. 196. The ideal pH range

for an indicator is 4.5–9.5.
12
10 Determination of the equivalence point
equivalence point final slope From the neutralisation curve (Fig. 25.2), the initial and final slopes are drawn
8
(Fig. 25.3) and a parallel line is drawn such that the mid-point is on the curve.
pH
6 This is the equivalence point, producing a titration value of x mL.

initial slope
4
2 Example calculations
0 Standardisation of a sodium hydroxide solution
0 50 100 150 200
volume of NaOH added (mL)
Sodium hydroxide solution (25.00 mL) is titrated with a standard solution of
hydrochloric acid (21.0 mL; 0.100 M). Calculate the molarity of the sodium
Fig. 25.3 Determination of the equivalence hydroxide solution.
point. Following the sequence in Box 24.1:
1. Write the equation:
Subscripts – ‘aq’ is used to represent
NaOH(aq) + HCl(aq) S NaCl(aq) + H2O(l)[25.2]
aqueous, ‘l’ the liquid state.
2. Equation [25.2] shows that 1 mole of NaOH requires 1 mole of HCl for
neutralisation, i.e. 1 mole of NaOH is equivalent to 1 mole of HCl.
3. You are trying to find the concentration of NaOH in mol L-1.
4. 1000 mL of HCl (1.0 M) K 1.0 mol NaOH
But the HCl is only 0.1 M
Self-test: is 0.1 M HCl equivalent to more or less moles of NaOH than 1.0 M
HCl?
Answer: less, therefore you must MULTIPLY by 0.1
1000 mL of HCl (0.1 M) K 0.1 mol L-1 * 1.0 mol NaOH
Self-test: is 1.0 mL of 0.1 HCl equivalent to more or less NaOH than 1000 mL?
Answer: less, therefore you must DIVIDE by 1000
0.1 mol L-1 * 1.0

1.0 mL of HCl (0.1 M) K mol NaOH
1000 mL
But the titration used 21.0 mL of 0.1 M HCl

Self-test: is 21.0 mL of 0.1 HCl equivalent to more or less NaOH than 1.0 mL?
Answer: more, therefore you must MULTIPLY by 21.0
0.1 mol L-1 * 21.0 mL * 1.0

21.0 mL of HCl (0.1 M) K mol NaOH
1000 mL
Since 25.00 mL of NaOH solution are equivalent to 21.0 mL of 0.1 M HCl
25.00 mL of NaOH solution 0.1 mol L-1 * 21.0 mL * 1.0

K mol NaOH
must contain 1000 mL

Do the self-test

K mol NaOH
must contain 1000 mL * 25.00 mL
Do the self-test
1000 mL of NaOH solution must contain
1000 mL * 0.1 mol L-1 * 21.0 mL * 1.0
K mol NaOH
1000 mL * 25.00 mL
Concentration of NaOH solution = 0.084 mol L−1
Standardisation of a sodium hydroxide solution using potassium

hydrogen phthalate (KHP) as a primary standard (p. 192)
An accurately weighed amount (5.1100 g) of potassium hydrogen phthalate
(KHC8H8O4) was dissolved in water (250.00 mL). This solution (25.00 mL)
required a sodium hydroxide solution (23.5 mL) for equivalence. What is the
molarity of the sodium hydroxide solution?
Following the sequence in Box 24.1:
1. Write the equation:
NaOH(aq) + KHC8H8O4(aq) S NaCl(aq) + H2O(l)[25.3]

2. Equation [25.3] shows that 1 mole of NaOH requires 1 mole of KHP for
neutralisation, i.e. 1 mole of NaOH is equivalent to 1 mole of KHP.
3. You are trying to find the concentration of NaOH in mol L-1.
4. Calculate the concentration of KHP from equation [11.1]:
Mass KHP/Mr
[C] =
Volume of solution
5.1100/204.22
5. [C] = = 0.100 mol L-1
250.00
6. 1000 mL of KHP (1.0 M) K 1.0 mol NaOH
But the KHP is only 0.1 M
Self-test: is 0.1 M KHP equivalent to more or less moles of NaOH than 1.0 M
KHP?
Answer: less, therefore you must MULTIPLY by 0.1
1000 mL of KHP (0.1 M) K 0.1 mol L-1 * 1.0 mol NaOH
Self-test: is 1.0 mL of 0.1 KHP equivalent to more or less NaOH than 1000 mL?
Answer: less, therefore you must DIVIDE by 1000
0.1 mol L-1 * 1.0

1.0 mL of KHP (0.1 M) K mol NaOH
1000 mL
But the titration used 25.00 mL of 0.1 M KHP

Self-test: is 25.00 mL of 0.1 KHP equivalent to more or less NaOH than 1.0 mL?
Answer: more, therefore you must MULTIPLY by 25.00
0.1 mol L-1 * 25.00 mL * 1.0

25.00 mL of KHP (0.1 M) K mol NaOH
1000 mL
Since 23.5 mL of NaOH solution are equivalent to 25.00 mL of 0.1 M KHP

K mol NaOH
must contain 1000 mL
Do the self-test

K mol NaOH
must contain 1000 mL * 23.5 mL
Do the self-test
1000 mL of NaOH solution must contain
1000 mL * 0.1 mol L-1 * 25.00 mL * 1.0
K mol NaOH
1000 mL * 23.5 mL
Concentration of NaOH solution = 0.106 mol L−1

De Levie, R. (2001) Aqueous Acid-Base Equilibria and Mendham, J., Denney, R.C., Barnes, J.D. and Thomas,
Titrations. Oxford University Press, Oxford. M.J.K. (2000) Vogel’s Textbook of Quantitative Chemi-
Harris, D.C. (2010) Quantitative Chemical Analysis, 8th
Chemical Analysis. Pearson, Harlow, Essex.
Jander, G., Jahr, K-F., Schulze, G. and Simon, J. (2009)
Volumetric Analysis, 17th edn. Walter de Gruyter Inc., Skoog, D.A., West, D.M., Holler F.J. and Crouch, S.R.
Germany. (2014) Fundamentals of Analytical Chemistry, 9th edn.
Brooks Cole, Belmont, CA.
McPherson, P. (2014) Practical Volumetric Analysis. RSC,
Cambridge.
Study exercise
25.1 An accurately weighed amount (3.6284 g) of crystallisation (x) in the original hydrated sodium
hydrated sodium carbonate (Na2CO3 x H2O) carbonate.
was dissolved in water (250.00 mL). An aliquot
(HINT: you need to calculate Mr for the original
(25.00 mL) of this solution required hydrochloric
Na2CO3 x H2O.)
acid solution (25.4 mL; 0.100 M) for equivalence.
Calculate the number of molecules of water of

26 Complexometric titrations
Complexometric titrations are mainly used to determine the concentration of

cations in solution. The method is based on the competition between a metal
ion (for example) and two ligands, one of which acts as an indicator and the
other is a component of a standard solution.
Some knowledge of the principles of metal-ligand binding is required in
order to understand this method.
Types of ligand
Ligands are chemical species that co-ordinate with metal ions to form a com-
CH2 3+ plex. They are classified on the basis of the number of points of attachment to
NH2 CH2
H2C H2N
the central ion.
NH2
Co ●● Monodentate ligand – here the ligand is bound to the central ion at only one
NH2
H2C H2N point, e.g. H2O, NH3.
H 2N CH2
H2C ●● Bidentate ligand – this has two points of attachment to the central ion,
e.g. ethylenediamine (en) (Fig. 26.1).
Fig. 26.1 Structure of [Co(en)3]3+. It is a
six-co-ordinate octahedral complex of ●● Multidentate ligand – these have several points of attachment, e.g. ethylene
ethylenediamine (en) with cobalt (III). The diaminetetra-acetic acid (EDTA), which is a hexadentate ligand (six points
complex has three five-membered rings. of attachment) (Fig. 26.2).
HOOC CH2 CH2 COOH The basis of a complexometric titration involving EDTA
N CH2 CH2 N
HOOC CH2 CH2 COOH The metal ion under investigation is bound to an indicator in solution (under
(a)
strict pH control). This solution is then titrated against a standard solution of
EDTA. This can be expressed in the form of an equation:
Metal – indicator + EDTA S metal – EDTA + indicator[26.1]

-
OC 2
O CH2 For example, if the indicator being used was solochrome black, the metal –
OC CH2 indicator solution would be red while the colour of the free indicator would be
O N CH2 blue (in the pH range 7–11). The reaction takes place if the EDTA displaces the
Pb indicator from the metal–indicator complex. Therefore the metal–EDTA com-
O N CH2
plex must be more stable thermodynamically than the metal–indicator complex.
OC CH2
O CH2 Stability of complexes
OC
The thermodynamic stability of a species is an indication of the extent to which
(b) that species will be formed (under certain conditions and provided that it is
allowed to reach equilibrium).
Fig. 26.2 Structure of EDTA. (a) EDTA con-
As an example, consider the general case of a metal, M, in solution together
tains two donor N atoms and four donor
with a monodentate ligand, L. It is possible to describe this system in terms of
O atoms. It can therefore form a hexaden-
tate complex (b) with a metal ion, e.g. Pb2+. step-wise equilibria:
M + L = ML K1 = [ML]/[M][L][26.2]
ML + L = ML 2 K2 = [ML 2]/[ML][L][26.3]
Or, in general terms:
ML (n - 1) + L = ML n Kn = [ML n]/[ML (n - 1)][L][26.4]
where K1, K2, c Kn are step-wise stability constants.

Complexometric titrations
An alternative approach for expressing the equilibria might be as follows:
M + L = ML b1 = [ML]/[M][L][26.5]
M + L 2 = ML 2 b2 = [ML 2]/[M][L2][26.6]
Or, in general terms:
Table 26.1 Stability constants of selected
metal–EDTA complexes (expressed as M + L n = ML n bn = [ML n]/[M][L]n[26.7]
log K)*
where b1, b2, ..., bn are the overall stability constants and are related to the
Ion log k Ion log k step-wise stability constants as follows:
Mg2+ 8.7 Ni2+ 18.6
bn = K1 * K2 * Kn[26.8]
Ca2+ 10.7 Cu2+ 18.8
14.3 21.9
Factors influencing the stability of complexes
Fe2+ Hg2+
The stability of a complex is related to the ability of the metal ion to complex
Co2+ 16.3 Sc3+ 23.1
with a given ligand, and to the characteristics of the ligand.
Al3+ 16.3 Cr3+ 24.0 End-points can be determined more easily when a single complex is
Cd2+ 16.6 In3+ 24.9 formed rather than when the complex is formed in a step-wise fashion. This
can be achieved by using the aminopolycarboxylic acid, EDTA (Fig. 26.2).
*Ionic strength of solution was 0.1 at 20 °C. In equations, EDTA can be expressed as H4Y. The disodium salt Na 2H2Y
Adapted from Vogel’s Textbook of Quantita-
is frequently used as a source of the complex-forming ion, H2Y2- . Thus the
tive Inorganic Analysis, 4th edn, J. Bassett,
R.C. Denney, G.H. Jeffery and J. Mendham, typical reaction of EDTA with a metal ion can be written in the following form:
Longman Scientific and Technical, Harlow,
(1978) p. 264. M2+ + H2Y2- S MY2- + 2H + [26.9]
The reaction of a metal ion with EDTA is always in the ratio 1:1. The stability
constants of selected metal–EDTA complexes are given in Table 26.1.
The detection of the end-point in titrations involving EDTA is most com-
monly achieved using a metal-ion indicator, i.e. a compound that changes its
colour when it complexes with a particular metal ion. The structures of selected
metal-ion indicators are shown in Fig. 26.3 and the properties of a variety of
metal-ion indicators are given in Table 26.2.
Types of complexometric titration

Direct titration
In this case, the metal ion is titrated with a standard solution of EDTA. The
solution containing the metal ion is buffered to an appropriate pH at which the
stability constant of the metal–EDTA complex is large. The free indicator has
a different colour from that of the metal–indicator complex.
OH OH
+
Na-O3S N N Back titration
NO2
In certain circumstances a particular metal ion cannot be titrated directly. This
includes situations where:
Solochrome black
(eriochrome black T)
●● The metal ion precipitates in the absence of EDTA.
OH OH ●● The metal ion reacts too slowly with EDTA.
N N SO-3 H+
CH3
●● The metal ion forms an inert complex.
Calmagite ●● No suitable indicator is available.
Fig. 26.3 Examples of metal-ion indicators: In these cases, a back titration is required. This involves addition of a known
solochrome black and calmagite. excess of EDTA to the metal ion (buffered to an appropriate pH). Then, the

Table 26.2 Properties of selected indicators
Colour of free
Indicator indicator Colour of metal–ion complex
Murexide
6 pH 9 (H4In- ) Red–violet orange (Cu2+ ), yellow (Ni2+ and Co2+ )
pH 9–11 (H3In2- ) Violet and red (Ca2+ )
7 pH 11 (H2In3- ) Blue
Solochrome black
6 pH 5 (H2In- ) Red In pH range 7–11 colour change is
pH 7–11 (HIn2- ) Blue blue–red (Mg, Mn, Zn, Cd, Hg, Pb, Cu,
7 pH 11.5 (In3- ) Orange Al, Fe, Ti, Co, Ni and Pt metals)
Calmagite
6 pH 5 (H2In- ) Red Same colour change as solochrome
pH 7–9 (HIn )2- Blue black but clearer and sharper
7 pH 11.4 (In3- ) Red–orange
Pyrocatechol violet
6 pH 1.5 (H4In) Red In pH range 2–6, yellow to blue
-
pH 2–6 (H3 In ) Yellow (Bi and Th); pH 7 violet to blue
pH 7 (H2In2- ) Violet (Cu2+ , Zn2+ , Cd2+ , Ni2+ and Co2+ )
7 pH 10 (In4- ) Blue
excess EDTA is titrated with a standard solution of a different metal ion. The
choice of a second metal ion is important as it must not displace the analyte
metal ion from its EDTA complex.
Practical considerations
●● pH adjustment is critical in EDTA titrations. The pH is monitored with a pH

meter or pH test paper.
●● The metal ion under investigation should ideally be approximately 0.25 mM
in a volume of 50–150 mL of solution. Dilution of the metal ion may be
necessary to avoid end-point detection problems.
●● Do not add excess indicator, as too intense a colour can lead to problems,
e.g. masking of the colour change.
●● It is sometimes difficult to detect the end-point because the colour change can
be slow to develop. Stirring is recommended to assist colour transformation.
●● The use of metal-ion indicators to indicate the end-point of complexomet-
ric titrations is based on a specific colour change. Some individuals may
Metal-ion indicators – For a metal-ion
indicator to be useful it must be less sta-
find it difficult to detect a particular colour change (e.g. those with col-
ble than the corresponding metal–EDTA
our blindness). Alternative approaches for end-point detection are available
complex.
based on a colorimeter/spectrophotometer (devices for measuring colour, see
Chapter 48) or electrochemical detection (see Chapter 54).

Example calculation
A solution of Ni2+ (25.0 mL) was titrated with the disodium salt of EDTA
(0.1036 mol L-1) at pH 5 and required 20.25 mL for the metal-indicator to
change colour. What is the concentration (g L-1) of the Ni2+ solution? The Ar
of nickel is 58.71 g mol-1.
1. Write the balanced equation
Ni2+ + H2EDTA2- S NiEDTA2- + 2H + [26.10]
2. Determine the equivalences
1mole of H2EDTA2- is equivalent to 1 mole of Ni2+
3. You now have a choice:

(i) do the calculation in moles and then convert to g L-1
or
(ii) working
The calculation below illustrates (ii).
4. 1000 mL of H2EDTA (1.0 M) K 58.71 g Ni2+
But H2EDTA is 0.1036 M
Self-test: is 0.1036M H2EDTA equivalent to more or less Ni 2+ than 1.0 M
H2EDTA?
Answer: less, therefore you must MULTIPLY by 0.1036.
1000 mL of H2EDTA (0.1036 M) K 0.1036 mol L-1 * 58.71 g mol-1 Ni2+
Self-test: is 1.0 mL 0.1036 M H2EDTA equivalent to more or less Ni 2+ than

1000 mL 0.1036 M H2EDTA?
Answer: less, therefore you must DIVIDE by 1000.
1.0 mL of H2EDTA (0.1036 M)
1.0 mL * 0.1036 mol L-1 * 58.71 g mol-1 2+
K Ni
1000 mL
But the titration used 20.25 mL of H2EDTA (0.1036 M)
Self-test: is 20.25 mL 0.1036 M H2EDTA equivalent to more or less Ni 2+ than
1.0 mL 0.1036 M H2EDTA?
Answer: more, therefore you must MULTIPLY by 20.25.
20.25 mL of H2EDTA (0.1036 M)
20.25 mL * 1.0 mL * 0.1036 mol L-1 * 58.71 g mol-1 2+
K Ni
1000 mL
Since 25.00 mL of Ni2+ solution are equivalent to 20.25 mL of 0.1036 M
H2EDTA solution
25.00 mL of Ni2+ solution contains
20.25 mL * 1.0 mL * 0.1036 mol L-1 * 58.71 g mol-1 2+
Ni
1000 mL

Do the self-test
1.00 mL of Ni2+ solution contains
20.25 mL * 1.0 mL * 0.1036 mol L-1 * 58.71 g mol-1 2+
Ni
25.00 mL * 1000 mL
Do the self-test
1000 mL of Ni2+ solution contains
1000 mL * 20.25 mL * 1.0 mL * 0.1036 mol L-1 * 58.71 g mol-1 2+
Ni
1.0 mL * 25.00 mL * 1000 mL
Concentration of Ni2+solution = 4.927 g L−1

Christian, G.D., Dasgupta, P.K. and Schug, K.A. (2014) Mendham, J., Denney, R.C., Barnes, J.D. and Thomas,
Analytical Chemistry, 7th edn. John Wiley & Sons Ltd, M.J.K. (2000) Vogel ’s Textbook of Quantitative
Chichester. Chemical Analysis, 6th edn. Prentice Hall, Harlow,
Harris, D.C. (2010) Quantitative Chemical Analysis, 8th Essex.
metric Analysis, 17th edn. Walter de Gruyter Inc., Germany. Skoog, D.A., West, D.M., Holler, F.J., and Crouch, S.R.
McPherson, P. (2014) Practical Volumetric Analysis. RSC, (2014) Fundamentals of Analytical Chemistry, 9th edn.
Cambridge. Brooks Cole, Belmont, CA.
Study exercise
26.1 You are the analyst working in a hospital labo- calcium oxalate which was filtered, washed
ratory. A urine sample from a patient must be and re-dissolved in acid. To this solution EDTA
analysed for Ca2+ and Mg2+ ions by the follow- (25.00 mL; 0.05 M) was added and the result-
ing procedure. The urine sample was diluted ing solution buffered to pH = 10 and titrated to
to exactly 2.000 L. After buffering to pH = 10, equivalence with an MgCl2 solution (10.00 mL;
a sample (25.00 mL) required EDTA (24.00 mL; 0.0500 M). From these results, calculate the
0.0500 M) for equivalence. In a second sam- amount of Ca2+ and Mg2+ (in mg) in the 2.000 L
ple (50.00 mL), the Ca2+ was precipitated as solution.

27 Redox titrations
All reduction–oxidation reactions involve a transfer of electrons. The oxidising

agent accepts electrons, and the reducing agent donates electrons. To establish
the reaction for a redox titration it is necessary to determine the ‘half-equation’
for both the oxidising agent and the reducing agent. By adding the two
‘half-equations’ it is possible to determine the overall equation for the titration
(Box 11.4). The basic theory of electrochemistry is described in Chapter 34.
One of the most common oxidants is potassium permanganate which in
acidic solution can undergo the following reaction:
MnO4- + 8H + + 5e - = Mn2+ + 4H2O[27.1]
Unfortunately, potassium permanganate is not obtainable in high enough purity

and can undergo decomposition by exposure to sunlight. Therefore it cannot be
used as a primary standard (p. 192). However, it can be used in redox titrations
provided it is standardised with sodium oxalate (which is available in high
purity). The redox reaction involving oxalate is as follows:
-
C2O2-
4 = 2CO2 + 2e [27.2]
The overall reaction between permanganate and oxalate can be obtained by

balancing the electrons on each side of the equation. This can be achieved by
multiplying eqn [27.1] by 2 and eqn [27.2] by 5, and then combining them as
follows:
2MnO4- + 16H + + 5C2O2-

4 = 2Mn
2+
+ 10CO2 + 8H2O[27.3]
Another common method for the standardisation of potassium permanganate

is to use iron (II):
Fe2+ = Fe3+ + e - [27.4]
The combined equation is obtained by multiplying eqn [27.4] by 5 and adding

to eqn [27.1]:
MnO4- + 8H + + 5Fe2+ = Mn2+ + 5Fe3+ + 4H2O[27.5]
Table 27.1 Common oxidising and reducing Potassium permanganate has a major advantage when used for titrations in that
agents used in redox titrations it can act as its own indicator.
A list of other common oxidising agents and reducing agents is given in
Oxidising agents Reducing agents Table 27.1.
Ceric Arsenite Many volumetric procedures involving redox reactions comprise multiple
Ce4+ AsO3-
3
stages which can lead to confusion in your approach to calculation. Typical of
Cr2O2-
7
Dichromate Fe2+ Ferrous these are experiments involving potassium iodate (KIO3) in which the iodate
H2O2 Hydrogen NH2OH Hydroxylamine is reacted with an excess of potassium iodide (KI) in acid solution and then the
peroxide iodine released is titrated with sodium thiosulphate Na 2S2O3.
IO3- Iodate Sn2+ Stannous Using the principles outlined in Boxes 11.4 and 11.5 you can work out a
direct relationship between KIO3 and Na 2S2O3 as shown below:
MnO4- Perman- S2O2-
3
Thiosulphate
ganate
IO3- + I - S I2 + H2O[27.6]

Redox titrations
Using the Boxes 11.4 and 11.5, identify the two partial ionic equations and
process them
10e + 12H + + 2IO3- S I2 + 6H2O[27.7]

10I - S 5I2 + 10e[27.8]
Add eqns [27.7] and [27.8] and divide by 2
IO3- + 5I - + 6H + S 3I2 + 3H2O[27.9]
Now look at the reaction between I2 and Na 2S2O3 using the same methodology;
2e + I2 S 2I - [27.10]
2S2O32- S S4O62- + 2e[27.11]
Add eqns [27.10] and [27.11]
2S2O32- + I2 S 2I - + S4O62- [27.12]
Now balance I2 between eqns [27.9] and [27.12] by multiplying [27.12] by 3

-
6S2O2- 2-
3 + 3I2 S 6I + 3 S4O6 [27.13]
Now combining eqns [27.9] and [27.13] you can see that:
1 mole of KIO3 is equivalent to 6 moles of Na2S2O3
Example calculation
Standardisation of potassium permanganate with a primary
standard, sodium oxalate
An accurately weighed portion of sodium oxalate (0.1550 g) was dissolved in
dilute sulphuric acid (250 mL). While maintaining the temperature of the solu-
tion above 70 °C, it was titrated to equivalence with potassium permanganate
solution (18.5 mL). What is the molarity of potassium permanganate?
1. Write the balanced equation for the reaction between the standard and the
test substance (using the two half-equations [27.1] and [27.2]):
2MnO4- + 16H + + 5C2O24 - = 2Mn2 + + 10CO2 + 8H2O[27.3]
2. Determine the equivalences of the reacting species:
2 moles of MnO4- are equivalent to 5 moles of C2O2-

4 .
3. Calculate the number of moles of standard substance (sodium oxalate)

used to reach the end-point of the reaction.
The molecular weight of sodium oxalate is 134 g mol-1. Therefore
0.1550 g of sodium oxalate is equivalent to:
0.1550 g
= 1.157 * 10-3 mol
134 g mol-1
4. Calculate the corresponding number of moles of potassium permanganate
present in the volume of titrant added.

Redox titrations
From the equation:
5 moles of Na 2C2O4 = 2 moles of KMnO4
Therefore,
2
1 mole of Na 2C2O4 K moles of KMnO4
5
Therefore,
1.157 * 10-3 * 2
1.157 * 10-3 moles of Na 2C2O4 K moles of KMnO4
5
Therefore,
1.157 * 10-3 * 2
18.5 mL of KMnO4 solution contain moles of KMnO4
5
Therefore,
1000 mL of KMnO4 solution contain
1000 mL * 1.157 * 10 -3 * 2
moles of KMnO4
18.5 mL * 5
Concentration of KMnO4 solution = 0.025 mol L− 1

Jander, G., Jahr, K.F., Schulze, G. and Simon, J. (2009) Chemical Analysis. Pearson, Harlow, Essex.
Volumetric Analysis, 17th edn. Walter de Gruyter Inc., Skoog, D.A., West, D.M., Holler, F.J. and Crouch, S.R.
Germany. (2014) Fundamentals of Analytical Chemistry, 9th edn.
Mendham, J., Denney, R.C., Barnes, J.D. and Thomas, Brooks Cole, Belmont, CA.
M.J.K. (2000) Vogel ’s Textbook of Quantitative
Study exercise
27.1 Dilute aqueous hydrogen peroxide is sold com- concentration (% w/v) of ‘10–Volume Hydrogen
mercially to the cosmetic and health care indus- Peroxide’. ‘10–volume Hydrogen Peroxide’ (25.00
tries as ‘10–Volume Hydrogen Peroxide’. This mL) was dissolved in water and made up to
means that one volume of solution will decom- 250.00 mL. This solution (25.00 mL) was titrated
pose to give ten volumes of oxygen at STP. with a KMnO4 solution (48 mL; 0.02 M) to a pale-
From the following analytical data calculate the pink end-point in the presence of dilute H2SO4.

28 Precipitation titrations
Precipitation is the term used to describe the process whereby a substance

Definition leaves solution rapidly, forming either a crystalline solid or amorphous solid
(the precipitate). In the case of a precipitation titration, this process occurs
Argentimetric is derived from the Latin when the analyte forms a precipitate with the titrant. The most common types
argentum, which means silver. of precipitation titrations use silver nitrate as the titrant. They are often referred
to as argentimetric titrations.
Titration curves used in precipitation reactions usually use a c oncentration-
Definition dependent variable called the ‘p function’ rather than the concentration itself. The
p function for a species X is defined as follows:
To be exact, the p function should be
defined in terms of activities instead of pX = -log10 [X][28.1]
concentrations.
For example, in the titration of 100 mL of 0.1 mol L-1 NaCl with
0.1 mol L-1 AgNO3 the initial concentration of [Cl- ] is 0.1 mol L-1, so by using
eqn [28.1] the p function is 1 or pCl- = 1.
Titration curves – plots of concentration
When 25 mL of 0.1 mol L-1 AgNO3 has been added, 75 mL of NaCl
of titrand against volume of titrant used.
remains in a total volume of 125 mL. Therefore, the concentration of the chlo-
ride ion is given by
Table 28.1 Titration of 100 mL of 0.1 M
75 mL * 0.1 mol L-1
NaCl with 0.1 M AgNO3. (Note that Ks, for [Cl- ] = = 6 * 10-2 mol L-1[28.2]
AgCl = 1.1 * 10-10) 125 mL
0.1 M AgNO3 (mL) p Cl − pAg −

and pCl- = 1.22. (Note that the solubility product, Ks, of AgCl is 1.1 * 10-10,
see p. 106.)
0 0.1 0.0 Therefore:
25 1.2 8.7
50 1.5 8.5 [Ag + ] * [Cl- ] = Ks = 1.1 * 10-10[28.3]
90 2.3 7.7
95 2.6 7.4 or
98 3.0 7.0
99 3.3 6.7 pAg + + pCl- = 9.96 = pAgCl[28.4]
99.5 3.6 6.4
99.8 4.0 6.0
It was found above that pCl- = 1.22, hence pAg + = 9.96 - 1.22 = 8.74. In
a similar manner, the pAg + values can be calculated.
99.9 4.3 5.7
At the equivalence point [Ag + ] = [Cl- ]. Therefore:
100 5.0 5.0
100.1 5.7 4.3
[Ag + ] = [Cl- ] = 2Ks = 21.1 * 10-10 = 1.05 * 10-5[28.5]
100.2 6.0 4.0
100.5 6.4 3.6 pAg + = -log(1.05 * 10-5) = 4.98[28.6]
101 6.7 3.3
102 7.0 3.0
Beyond the equivalence point the situation changes. For 100.1 mL AgNO3
solution:
105 7.4 2.6
110 7.7 2.3 0.1 mL * 0.1 mol L-1
120 8.0 2.0 [Ag + ] = = 5 * 10-5 mol L-1[28.7]
200.1 mL
130 8.1 1.9
140 8.2 1.8 or pAg + = 4.3. Therefore,
150 8.3 1.7
pCl- = pAgCl - pAg + = 9.96 - 4.3 = 5.66
Adapted from Vogel’s Textbook of Q uantitative
Inorganic Analysis, 4th edn, J. Bassett, R.C.
Denney, G.H. Jeffery and J. Mendham, Longman Values calculated in this way up to the addition of 150 mL of 0.1 mol L-1 AgNO3
Scientific and Technical, Harlow (1978), p. 280. are given in Table 28.1 and the titration curve in Fig. 28.1.

Precipitation titrations
equivalence point, pAg+ = 5 End-point determination

10.0
Three techniques are commonly used to determine the end-point in precipita-
8.0 tion titrations. They are:
6.0
pAg+
1. potentiometric methods;
4.0
2. chemical indicator methods;
2.0
3. light-scattering methods, exemplified by turbidimetry or nephelometry.
0.0
0 50 100 150
Only indicator methods will be discussed further. Three types of indicator
0.1 M AgNO3 (mL)
methods can be applied to determine the end-point of an Ag + and a Cl- titra-
Fig. 28.1 Precipitation titration curve. Initial tion. These are:
and final slopes are drawn (see Fig. 25.3)
1. Mohr titration, which involves the formation of a coloured precipitate by
and a parallel line is drawn such that the
mid-point is on the curve. This is the equiv- reaction with the indicator. For example, in the determination of chloride
alence point. concentration with silver nitrate a small amount of potassium chromate
solution is added as an indicator. This results in the formation of a red
silver chromate (Ag2CrO4) precipitate at the end-point:
In acid media (pH 6 6), the concentra-
2Ag + + CrO2-
4 S Ag 2CrO4(s)
tion of CrO2-
4 is lowered by the following [28.8]
reaction: CrO2- + - 2- (red)
4 + 2H 2HCrO4 Cr2O7 +
H2O. In a lkaline media (pH 7 10),
Ag(OH)(s) may precipitate. In this case, the precipitate may form slightly after the end-point, but this
error can usually be neglected. Also, the titration should be done in neutral
or slighly alkaline solution (pH 6.5–9) otherwise silver chromate might
not be formed.
2. Volhard titration, which involves the formation of a soluble coloured com-
pound. This approach is exemplified by the quantitative analysis of chlo-
rides, bromides and iodides by back titration. In this case, the halide is
titrated with silver:
In all argentimetric titrations, strong
Ag + + Cl- S AgCl(s)[28.9]
light (including daylight) should be
avoided as it can lead to decomposition
of the silver salts.
Excess silver ions are then titrated with standard potassium thiocyanate
solution in the presence of an iron (III) salt:
Ag + + SCN- S AgSCN(s)[28.10]
When all the Ag + has been reacted, the SCN- reacts with Fe3+ to form a
red complex, indicating the end-point:
Fe3+ + SCN- S FeSCN2+

[28.11]
(red)
A problem with the determination of chloride by this approach is that

the end-point coloration slowly fades, as AgCl is more soluble than
- O O
AgSCN. As a consequence the AgCl slowly dissolves to be replaced by the
O
FeSCN2+ . Two approaches are possible to prevent this secondary reaction
Cl Cl from taking place. The most common method is to filter off the AgCl and
CO2- titrate only the Ag + left in solution. Alternatively, add a few millilitres of
an immiscible liquid (e.g. nitrobenzene) to the titrand prior to the back
titration. The nitrobenzene acts to ‘coat’ the AgCl precipitate, thereby iso-
Fig. 28.2 Structure of dichlorofluorescein. lating it from the SCN- .

Precipitation titrations
Table 28.2 Selected applications of precip- 3. Fajans titration, which involves the adsorption of a coloured indicator onto
itation titrations the precipitate at the end-point, resulting in a colour change. During this
adsorption process a change occurs in the indicator resulting in a change
Analyte Comments of colour. The indicators used for this are often anionic dyes, e.g. fluores-
Cl - , Br - Mohr method: Ag2CrO4 used cein or eosin. The most common indicator for AgCl is dichlorofluorescein
as end-point (Fig. 28.2) (this is greenish yellow in solution but changes colour to pink
Br - , I - , AsO4- Volhard method: precipitate when it is adsorbed on AgCl).
removal is unnecessary
Selected examples of precipitation titrations are shown in Table 28.2.
Cl - , CN- , Volhard method: precipitate
CO2- 3 removal is required
Cl - , Br - , I - , Fajans method: titration with
SCN- Ag+ . Detection with fluorescein,
dichlorofluorescein and eosin
F- Titration with Th(NO3)4 to
p
roduce ThF4. End-point
detection with alizarin red S
Adapted from: Quantitative Chemical Analysis,

4th edn, D.C. Harris, W.H. Freeman, New York
(1995), p. 176.

metric Analysis, 17th edn. Walter de Gruyter Inc., Germany. Skoog, D.A., West, D.M., Holler, F.J. and Crouch, S.R.
Mendham, J., Denney, R.C., Barnes, J.D. and Thomas, (2014) Fundamentals of Analytical Chemistry, 9th edn.
M.J.K. (2000) Vogel ’s Textbook of Quantitative Brooks Cole, Belmont, CA.
Study exercise
28.1 You are a major manufacturer of fireworks and ‘potassium nitrate’ (4.0124 g) was dissolved
you suspect that your bulk supplier of potas- in water and made up to 250.00 mL. This solu-
sium nitrate, a white crystalline solid, has tion (25.00 mL) required silver nitrate solution
been adulterating the potassium nitrate with (10.3 mL; 0.1 M) for equivalence using dichlor-
salt to increase his profits. Your analysis of the ofluorescein as indicator. Calculate the % (w/w)
‘potassium nitrate’ gave the following results: salt in the ‘potassium nitrate’.

Instrumental techniques
29. Basic spectroscopy 217
30. Atomic spectroscopy 226
31. X-ray fluorescence spectroscopy 243
32. Chromatography 250
33. Electrophoresis 287
34. Electroanalytical techniques 302
35. Using radioisotopes 309
36. Infrared and Raman spectroscopy 318
37. Nuclear magnetic resonance spectroscopy 332
38. Mass spectrometry 353
39. X-ray diffraction 362
40. Thermal analysis 370

29 Basic spectroscopy
The absorption and emission of electromagnetic radiation (Fig. 29.1) of

Spectroscopic techniques – can be
specific energy (wavelength) are characteristic features of many molecules,
used to:
involving the movement of electrons between different energy states, in
●● tentatively identify compounds, accordance with the laws of quantum mechanics. Electrons in atoms or mol-
by determining their absorption or ecules are distributed at various energy levels, but are mainly at the lowest
emission spectra; energy level, usually termed the ground state. When exposed to energy (e.g.
●● quantify substances, either singly or from electromagnetic radiation), electrons may be excited to higher energy
in the presence of other compounds, levels (excited states), with the associated absorption of energy at specific
by measuring the signal strength at
wavelengths giving rise to an absorption spectrum. One quantum of energy is
an appropriate wavelength;
●● determine molecular structure;
absorbed for a single electron transition from the ground state to an excited
●● follow reactions, by measuring the state. On the other hand, when an electron returns to its ground state, one
disappearance of a substance, or the quantum of energy is released; this may be dissipated to the surrounding
appearance of a product as a func- molecules (as heat) or may give rise to an emission spectrum. The energy
tion of time. change (∆E) for an electron moving between two energy states, E1 and E2,
is given by the equation:
106
∆E = E1 - E2 = hy[29.1]
104
radio where h is the Planck constant (p. 43) and y is the frequency of the electromag-
waves netic radiation expressed in Hz or s-1). Frequency is related to wavelength (l,
102
usually expressed in nm) and the speed of electromagnetic radiation, c (p. 43)
by the expression:
100
infrared microwaves
y = c , l[29.2]
800 770 nm 10-2
wavelength (nm)
red
infrared
700 10-4 UV/visible spectrophotometry
orange This is a widely used technique for measuring the absorption of radiation in the
wavelength (nm)
600 visible 10-6

yellow spectrum visible visible and UV regions of the spectrum. A spectrophotometer is an instrument
green ultraviolet designed to allow precise measurement at a particular wavelength, while a
500 10-8 colorimeter is a simpler instrument, using filters to measure broader wavebands
blue
violet x-rays
(e.g. light in the green, red or blue regions of the visible spectrum).
400 10-10
ultraviolet 390 nm
gamma rays
Principles of light absorption
300 10-12
Two fundamental principles govern the absorption of light passing through a
cosmic rays
solution:
10-14
●● The absorption of light is exponentially related to the number of molecules
Fig. 29.1 The electromagnetic spectrum. of the absorbing solute that are encountered, i.e. the solute concentration [C].
●● The absorption of light is exponentially related to the length of the light path
through the absorbing solution, l.
Definitions
These two principles are combined in the Beer–Lambert relationship (some-
Absorbance (A) – this is given by: times referred to simply as ‘Beer’s Law’), which is usually expressed in terms
A = log10(I0/I). of absorbance (A) – the logarithm of the ratio of the incident light (I0) to the
emergent light (I):
Usually shown as Ax where ‘x ’ is the
wavelength, in nanometres.
A = el[C][29.3]

Basic spectroscopy
where A is absorbance, e is a constant for the absorbing substance and the wave-
Definition length, termed the absorption coefficient or absorptivity, and [C] is expressed
either as mol L-1 or g L-1 (see p. 102) and l is given in cm. This relationship
Transmittance (T) – this is usually is extremely useful, since most spectrophotometers are constructed to give a
expressed as a percentage, where direct measurement of absorbance (A), sometimes also termed extinction (E),
T = (I/I0) * 100(%) of a solution (older texts may use the outdated term optical density, OD). Note
that for substances obeying the Beer–Lambert relationship, A is linearly related
to [C]. Absorbance at a particular wavelength is often shown as a subscript
(e.g. A550 represents the absorbance at 550 nm). The proportion of light passing
through the solution is known as the transmittance (T), and is calculated as the
ratio of the emergent and incident light intensities.
Some instruments have two scales:
●● an exponential scale from zero to infinity, measuring absorbance;
●● a linear scale from 0 to 100, measuring (per cent) transmittance.
For most practical purposes, the Beer–Lambert relationship applies and you
should use the absorbance scale.
sample
cuvette UV/visible spectrophotometer
monochromator The principal components of a UV/visible spectrophotometer are shown in
Fig. 29.2. High intensity tungsten bulbs are used as the light source in basic
instruments, capable of operating in the visible region (i.e. 400–700 nm).
light detector amplifier/
exit test readout
Deuterium lamps are used for UV spectrophotometry (200–400 nm); these
source
slit solution lamps are fitted with quartz envelopes, since glass does not transmit UV
radiation.
Fig. 29.2 Components of a UV/visible
A major improvement over the simple colorimeter is the use of a dif-
spectrophotometer.
fraction grating to produce a parallel beam of monochromatic light from the
(polychromatic) light source. In practice the light emerging from such a mon-
ochromator is not of a single wavelength, but is a narrow band of wavelengths.
Safety Note Working with spec- This bandwidth is an important characteristic, since it determines the wave-
trophotometers – take care not to spill lengths used in absorption measurements – the bandwidth of basic spectro-
aqueous solutions into the inside of the photometers is around 5–10 nm while research instruments have bandwidths
instrument, owing to the risk of elec- of less than 1 nm.
tric shock during use (switch off at the Bandwidth is affected by the width of the exit slit (the slit width), since
mains electric immediately and seek the bandwidth will be reduced by decreasing the slit width. To obtain accurate
assistance if this should happen). data at a particular wavelength setting, the narrowest possible slit width should
be used. However, decreasing the slit width also reduces the amount of light
reaching the detector, decreasing the signal-to-noise ratio. The extent to which
the slit width can be reduced depends upon the sensitivity and stability of the
Using plastic disposable cuvettes – detection/amplification system and the presence of stray light.
These are adequate for work in the Most UV/visible spectrophotometers are designed to take cuvettes with an
near-UV region as well as the visible optical path length of 10 mm. Disposable plastic cuvettes are suitable for rou-
range. tine work in the visible range using aqueous and alcohol-based solvents, while
glass cuvettes are useful for other organic solvents. Glass cuvettes are manu-
factured to more exacting standards, so use optically matched glass cuvettes
Handling cells – never handle the cells for accurate work, especially at low absorbances ( 6 0.1), where any differences
by the polished sides. in the optical properties of cuvettes for reference and test samples will be pro-
nounced. Glass and plastic absorb UV light and quartz cuvettes must be used
at wavelengths below 300 nm (see also Table 29.1).
Before taking a measurement, make sure that cuvettes are clean, unscratched,
dry on the outside, filled to the correct level and in the correct position in their
sample holders. Unwanted material can accumulate on the inside faces of glass/

Basic spectroscopy
Table 29.1 Cell materials for use in the UV/ quartz cuvettes, so remove any deposits using acetone on a cotton bud, or soak
visible spectral regions overnight in 1 mol L-1 nitric acid. Corrosive and hazardous solutions must be
used in cuvettes with tightly fitting lids, to prevent damage to the instrument and
Application Application for to reduce the risk of accidental spillage.
for UV region Visible region
Cell material 200–370 nm 370–850 nm
Basic instruments use photocells similar to those used in simple color-
imeters or photodiode detectors. In many cases, a different photocell must be
Silica U U used at wavelengths above and below 550–600 nm, due to differences in the
Quartz U U sensitivity of such detectors over the visible waveband. The detectors used in
Glass * U more sophisticated instruments give increased sensitivity and stability when
compared to photocells.
Plastic * U
Digital displays are increasingly used in preference to needle-type meters,
as they are not prone to parallax errors and misreading of the absorbance scale.
Some digital instruments can be calibrated to give a direct readout of the con-
Examples – the molar absorptivity centration of the test substance.
of phenol is 6.20 * 103 L mol-1 cm -1
at 210 nm. For a test solution giving
an absorbance of 0.21 in a cell with a
light path of 5 mm, using eqn [29.3] Key Point Basic spectrophotometers are most accurate
this is equal to a concentration of: within the absorbance range from 0.00 to 1.00 and your calibra-
0.21 = 6.20 * 103 * 0.5 * [C] tion standards and test solutions should be prepared to give
[C] = 0.000067.7 mol L-1 readings within this range.
(or 67.7 mmol L-1)
Types of UV/visible spectrophotometer

Safety Note Never balance full Basic instruments are single beam spectrophotometers in which there is only
cells on the instrument. one light path. The instrument is set to zero absorbance using a blank solu-
tion, which is then replaced by the test solution, to obtain an absorbance
reading. An alternative approach is used in double beam spectrophotometers,
where the light beam from the monochromator is split into two separate
beams, one beam passing through the test solution and the other through a
reference blank. Absorbance is then measured by an electronic circuit which
compares the output from the reference (blank) and sample cuvettes. Double
beam spectrophotometry reduces measurement errors caused by fluctuations
in output from the light source or changes in the sensitivity of the detection
system, since reference and test solutions are measured at the same time (Box
29.1). Recording spectrophotometers are double beam instruments, designed
A340
for use with a chart recorder, either by recording the difference in absorbance
between reference and test solutions across a predetermined waveband to give
an absorption spectrum (Fig. 29.3), or by recording the change in absorbance
at a particular wavelength as a function of time (e.g. in a kinetics experiment).
NADH Quantitative spectrophotometric analysis

NAD1
A single (purified) substance in solution can be quantified using the Beer–
Lambert relationship (eqn [29.3]), provided its absorptivity is known at a par-
ticular wavelength (usually at the absorption maximum for the substance, since
200 300 400 500 this will give the greatest sensitivity). The molar absorptivity is the absorb-
wavelength (nm)
ance given by a solution with a concentration of 1 mol L-1(= 1 kmol m -3) of
Fig. 29.3 Absorption spectra of nicotin- the compound in a light path of 1 cm. The appropriate value may be availa-
amide adenine dinucleotide in oxidised ble from tabulated spectral data (e.g. Anon., 1963), or it can be determined
(NAD +) and reduced (NADH) form. Note experimentally by measuring the absorbance of known concentrations of the
the 340 nm absorption peak (A340), used substance (Box 29.1) and plotting a standard curve (see Chapter 48). This
for quantitative work. should confirm that the relationship is linear over the desired concentration
range and the slope of the line will give the molar absorptivity.

Basic spectroscopy
Box 29.1 How to use a UV/visible spectrophotometer
1. Switch on and select the correct lamp for your meas- 8. Set the absorbance reading to zero: usually via a dial,
urements (e.g. deuterium for UV, tungsten for visible or digital readout.
light).
9. Analyse your samples: replace the appropriate refer-
2. Allow up to 15 min for the lamp to warm up and for ence blank with a test sample, allow the absorbance
the instrument to stabilise before use. reading to stabilise (5–10 s) and read the absorbance
3. Select the appropriate wavelength: on older value from the meter/readout device. For absorbance
instruments a dial is used to adjust the monochro- readings greater than 1 (i.e. 610% transmission), the
mator, while newer machines have microprocessor- signal-to-noise ratio is too low for accurate results.
controlled wavelength selection. Your analysis may require a calibration curve or you
may be able to use the Beer–Lambert relationship
4. Select the appropriate detector: some instruments (eqn [29.3]) to determine the concentration of test
choose the correct detector automatically (on the substance in your samples.
basis of the specified wavelength), while others have
manual selection. 10. Check the scale zero at regular intervals using a ref-
erence blank (e.g. after every 10 samples).
5. Choose the correct slit width (if available): this may
be specified in the protocol you are following, or may 11. Check the reproducibility of the instrument: measure
be chosen on the manufacturer’s recommendations. the absorbance of a single solution several times dur-
ing your analysis. It should give the same value.
6. Insert appropriate reference blank(s): single beam
instruments use a single cuvette, while double beam Problems (and solutions): inaccurate/unstable read-
instruments use two cuvettes (a matched pair for ings are most often due to incorrect use of cuvettes,
accurate work). The reference blanks should match for example dirt, fingerprints or test solution on out-
the test solution in all respects apart from the sub- side of cuvette (wipe the clear faces using a soft tissue
stance under test, i.e. they should contain all rea- before insertion into the cuvette holder and handle
gents apart from this substance. Make sure that the only by the opaque faces), condensation (if cold solu-
cuvettes are positioned correctly, with their polished tions aren’t allowed to reach room temperature before
(transparent) faces in the light path, and that they are use), air bubbles (which scatter light and increase the
accurately located in the cuvette holder(s). absorbance; tap gently to remove), insufficient solution
7. Check/adjust the 0% transmittance: most instruments (causing refraction of light at the meniscus), particulate
have a control which allows you to zero the detector material in the solution (check for ‘cloudiness’ in the
output in the absence of any light (termed ‘dark cur- solution and centrifuge before use, where necessary)
rent’ correction). Some microprocessor-controlled or incorrect positioning in light path (locate in correct
instruments carry out this step automatically. position).
The specific absorptivity is the absorbance given by a solution containing

Measuring absorbances in colorimetric
10 g L-1 (i.e. 1% w/v) of the compound in a light path of 1 cm. This is useful for
analysis – if any final solution has an
substances of unknown molecular weight (e.g. proteins or nucleic acids), where
absorbance that is too high to be read
the amount of substance in solution is expressed in terms of its mass, rather
with accuracy on your spectropho-
than as a molar concentration. For use in eqn [29.3], the specific absorptivity
tometer (e.g. A 7 2), it is bad practice
should be divided by 10 to give the solute concentration in g L-1.
to dilute the solution so that it can be
This simple approach cannot be used for mixed samples where several sub-
measured. This dilutes both the sample
stances have a significant absorption at a particular wavelength. In such cases,
molecules and the colour reagents to an
it may be possible to estimate the amount of each substance by measuring the
equal extent. Instead, you should dilute
absorbance at several wavelengths.
the original sample and reassay.
Fluorescence
With most molecules, after electrons are raised to a higher energy level by
absorption of electromagnetic radiation, they soon fall back to the ground
state by radiationless transfer of energy (heat) to the solvent. However, with

Basic spectroscopy
some molecules, the events shown in Fig. 29.4 may occur, i.e. electrons may
second lose only part of their energy by non-radiant routes and the rest may be
excited state
emitted as electromagnetic radiation, a phenomenon known as fluorescence.
non-radiant
loss (heat) Since not all of the energy that was absorbed is emitted (due to non-radiant
loss), the wavelength of the fluorescent light is longer than the absorbed light
energy level of electron
first (longer wavelength = lower energy). Thus, a fluorescent molecule has both
excited state an absorption spectrum and an emission spectrum. The difference between
absorption of light energy
non-radiant loss (heat)
the excitation wavelength (lex) and the emission wavelength (lem), measured
in nm, is known as the Stokes shift, and is fundamental to the sensitivity of
fluorescence
fluorescence techniques. The existence of a Stokes shift means that emitted

light can be detected against a low background, independently of the excita-
tion wavelength.
Most fluorescent molecules have the following features:
ground state
●● A highly conjugated system (alternating double and single bonds),
Fig. 29.4 Energy levels and energy transi- involving aromatic or heterocyclic rings, usually containing O or N (as
tions in fluorescence. heteroatoms).
●● A condensed system of fused rings, with one or more heteroatoms.
●● Electron-donating groups such as -OH, -OCH3, -NH2 and -NR2, together
(i)
with electron-attracting groups elsewhere in the molecule, in conjugation
HO C O C O
(iii) C C (ii) C C (iv)
with the electron-donating groups.
C C C C ●● A rigid, planar structure.
C C C
OH
C C Figure 29.5 illustrates many of these features for fluorescein, used in a range
C C of applications including visualisation of nucleic acids, fluorescent antibody
O
C C tests, etc.
C
* Fluorescence spectrophotometry
Fig. 29.5 Fluorescein – a widely used The principal components of a fluorescence spectrophotometer (fluorimeter)
fluorescent label, showing (i) a planar are shown in Fig. 29.6. The instrument contains two monochromators, one
conjugated system of fused rings; (ii) het- to select the excitation wavelength and the other to monitor the light emitted,
eroatoms within the conjugated structures; usually at 90° to the incident beam (though light is actually emitted in all
(iii) an electron-donating group; and (iv) an
directions). As an example, the wavelengths used to measure the highly fluo-
electron-attracting group. In fluorogenic
rescent compound aminomethylcoumarin are 388 nm (excitation) and 440 nm
enzyme substrates, linkage is usually via
one of the two hydroxyl groups, while in (emission). Some examples of molecules with intrinsic fluorescence are given
fluoroscein-labelled proteins, linkage is via in Table 29.2.
an isothiocyanate group (N “ C “ S) on the Compared with UV/visible spectrophotometry, fluorescence spectroscopy
lowermost ring (*) has certain advantages, including:
●● Enhanced sensitivity (up to 1000-fold), since the emitted light is detected

against a background of zero, in contrast to spectrophotometry where
Table 29.2 Examples of compounds with
small changes in signal are measured against a large ‘background’ (see
intrinsic fluorescence
eqn [29.3]).
Drugs ●● Increased specificity, because not one, but two specific wavelengths are
Aspirin, morphine, barbiturates, propanalol, required for a particular compound.
ampicillin, tetracyclines
However, there are also certain drawbacks:
Vitamins
Riboflavin, vitamins A, B6 and E, nicotinamide ●● Not all compounds show intrinsic fluorescence, limiting its application.
Pollutants However, some non-fluorescent compounds may be coupled to fluorescent
dyes, or fluorophores (e.g. alcohol ethoxylates may be coupled to naphthoyl
Naphthalene, anthracene, benzopyrene
chloride).

Basic spectroscopy
●●The light emitted can be less than expected due to quenching, i.e. when
light source
substances in the sample (e.g. oxygen) either interfere with energy transfer,
or absorb the emitted light (in some instances, the sample molecules may
self-quench if they are present at high concentration).
excitation The sensitivity of fluorescence has made it invaluable in techniques in
monochromator
or filter which specific antibodies are linked to a fluorescent dye, including:
●● fluorescence immunoassay (FIA);
sample cuvette ●● immunohistochemistry, which requires the use of a fluorescence microscope,

(test solution) for example using fluorescent antibodies, or fluorescent in situ hybridisation
(FISH) for nucleic acid detection.
Phosphorescence and luminescence

emission
monochromator A phenomenon related to fluorescence is phosphorescence, which is the
or filter emission of light following inter-system crossing between electron orbitals
detector
(e.g. between excited singlet and triplet states). Light emission in phospho-
rescence usually continues after the exciting energy is no longer applied and,
readout since more energy is lost in inter-system crossing, the emission wavelengths are
generally longer than with fluorescence. Phosphorescence has limited applica-
Fig. 29.6 Components of a fluorimeter (flu- tions in forensic science.
orescence spectrophotometer). Note that Luminescence (or chemiluminescence) is another phenomenon in which
sample cuvettes for fluorimetry must have light is emitted, but here the energy for the initial excitation of electrons is
clear sides all round. provided by a chemical reaction rather than by electromagnetic radiation. An
example is the action of the enzyme luciferase, extracted from fireflies, which
catalyses the following reaction:
Safety Note Caution is needed
when using strong (concentrated) luciferin + ATP + O2
acids – always work in a fume cupboard, 1 oxyluciferin + AMP + PPi + CO2 + light[29.4]
wear gloves to protect your hands from
acid burns, and always rinse affected The light produced is either yellow-green (560 nm) or red (620 nm). This sys-
areas with large amounts of water. tem can be used in the analysis of ATP (e.g. to determine ATP concentration in
a biological sample). Measurement can be performed using the photomultiplier
tubes of a scintillation counter to detect the emitted light, with calibration of
the output using a series of standards of known ATP content.
Safe working in atomic spectroscopy –
the use of high pressure gas cylinders
can be particularly hazardous. Always Atomic spectroscopy
consult a member of staff before using Atoms of certain metals will absorb and emit radiation of specific wavelengths
such apparatus. when heated in a flame, in direct proportion to the number of atoms present.
Atomic spectrophotometric techniques measure the absorption or emission
of particular wavelengths of UV and visible light, to identify and quantify
nebuliser amplifier/ such metals.
flame readout
compressed detector
air
Flame atomic emission spectrophotometry (or flame photometry)
filter or
The principal components of a flame photometer are shown in Fig. 29.7. A liquid
burner
monochromator sample is converted into an aerosol in a nebuliser (atomiser) before being intro-
test duced into the flame, where a small proportion (typically less than 1 in 10 000)
solution
of the atoms will be raised to a higher energy level, releasing this energy as light
Fig. 29.7 Components of a flame photometer. of a specific wavelength, which is passed through a filter to a photocell detector.

Basic spectroscopy
Box 29.2 How to use a flame photometer
1. Allow time for the instrument to stabilise. Switch on 4. Analyse all solutions in duplicate, so that repeatabil-
the instrument, light the flame and wait at least 5 min ity can be assessed.
before analysing your solutions.
5. Check your calibration. Make repeated measure-
2. Check for impurities in your reagents. For example, ments of a standard solution of known concentration
if you are measuring Na+ in an acid digest of some after every six or seven samples, to confirm that the
material, for e.g., soil, check the Na+ content of a instrument calibration is still valid.
reagent blank, containing everything except the soil,
processed in exactly the same way as the samples. 6. Consider the possibility of interference. Other metal
Subtract this value from your sample values to obtain atoms may emit light which is detected by the pho-
the true Na+ content. tocell, since the filters cover a wider waveband than
the emission line of a particular element. This can be a
3. Quantify your samples using a calibration curve. Cali- serious problem if you are trying to measure low con-
bration standards should cover the expected concen- centrations of a particular metal in the presence of high
tration range for the test solutions – your calibration concentrations of other metals (e.g. Na+ in sea water),
curve may be non-linear (especially at concentrations or other substances which form complexes with the
above 1 mmol L-1, i.e. 1 mol m -3). test metal, suppressing the signal (e.g. phosphate).
Flame photometry is used to measure the alkali metal ions K+ , Na+ and Ca++ in
biological fluids and water samples (Box 29.2).
When using a flame photometer:
●● Allow time for the instrument to stabilise. Switch on the instrument, light the
flame and wait at least 5 min before analysing your solutions.
●● Check for impurities in your reagents. For example, if you are measuring K+
in an acid digest of some biological material, check the K+ content of a rea-
gent blank, containing everything except the biological material, processed
in exactly the same way as the samples. Subtract this value from your sample
values to obtain the true K+ content. If using acid digestion, always work
within a fume hood and wear gloves and safety glasses at all times. Rinse
any spillages with a large volume of water.
●● Quantify your samples using a calibration curve (Chapter 48). Calibra-
tion standards should cover the expected concentration range for the test
solutions – your calibration curve may be non-linear (especially at concen-
trations above 1mmol L-1, i.e. 1 mol m -3 in SI units).
●● Analyse all solutions in duplicate, so that reproducibility can be assessed.
●● Check your calibration. Make repeated measurements of a standard solution
of known concentration after every six or seven samples, to confirm that the
instrument calibration is still valid.
Plotting calibration curves in quantita-
tive analysis – do not force your cali-
●● Consider the possibility of interference. Other metal atoms may emit light
bration line to pass through zero if it
which is detected by the photocell, since the filters cover a wider waveband
clearly does not. There is no reason to
than the emission line of a particular element. This can be a serious problem
assume that the zero value is any more
if you are trying to measure low concentrations of a particular metal in the
accurate than any other reading you
presence of high concentrations of other metals (e.g. Na+ in sea water), or
have made.
other substances which form complexes with the test metal, suppressing the
signal (e.g. phosphate).

Basic spectroscopy
Atomic absorption spectroscopy

This technique is applicable to a broad range of metal ions, including those
of Pb, Cu, Zn, etc. It relies on the absorption of light of a specific wavelength
by atoms dispersed in a flame. The appropriate wavelength is provided by a
hollow cathode lamp, coated with the element to be analysed, focused through
the flame and onto the monochromator detector. When the sample is introduced
into the flame, it will decrease the light detected in direct proportion to the
amount of metal present. Practical advantages over flame photometry include:
●● improved sensitivity;
●● increased precision;
●● decreased interference.
Variants of this method include flameless atomic absorption spectroscopy and
atomic fluorescence spectroscopy, both of which are more sensitive than the
flame atomic absorption technique. Chapter 30 gives further details on the
practical applications of atomic spectroscopy.
Text reference
Anon. (1963) Tables of Spectrophotometric Absorp- Detection of Elements (International Union of Pure and
tion Data for Compounds used for the Colorimetric Applied Chemistry). Butterworth-Heinemann, London.

Christian, G.D., Dasgupta, P.K. and Schug, K.A. (2014) McMohan, G. (2008) Analytical Instrumentation: A Guide
Analytical Chemistry, 7th edn. John Wiley & Sons Ltd, to Laboratory, Portable and Miniaturized Instruments.
Chichester. John Wiley & Sons Ltd, Chichester.
Gore, M.G. (ed.) (2000) Spectrophotometry and Spectro- Rouessac, F. and Rouessac, A. (2007) Chemical Analy-
fluorimetry: A Practical Approach, 2nd edn. Oxford sis: Modern Instrumentation Methods and Techniques,
University Press, Oxford. 2nd edn. John Wiley & Sons Ltd, Chichester.
Harris, D.A. (1996) Light Spectroscopy. Bios, Oxford. Schwedt, G. (1997) The Essential Guide to Analytical
Harris, D.C. (2010) Quantitative Chemical Analysis, Chemistry. 2nd edn. John Wiley & Sons Ltd, Chichester.
8th edn. Freeman, New York. Skoog, D.A., West, D.M., Holler, F.J. and Crouch, S.R.
Kellner, R., Mermet, J.M., Otto, M., Valcarcel, M. and (2014) Fundamentals of Analytical Chemistry, 9th edn.
Widmer, H.M. (2004) Analytical Chemistry: A Modern Brooks Cole, Belmont, CA.
Approach to Analytical Science, 2nd edn. John Wiley &

Basic spectroscopy
Study exercises
29.1 Write a protocol for using a spectrophotome- path length 5 mm, based on an absorptivity
ter. After reading this chapter, prepare a detailed of 20 L g -1 cm -1.
stepwise protocol explaining how to use one of
(b) The amount (ng) of pentachlorophenol in a
the spectrophotometers in your department. Ask
50 mL sub-sample from a test solution where
another student or a tutor to evaluate your pro-
A300 = 0.31 in a cuvette of path length 1 cm,
tocol and provide you with feedback.
based on an absorptivity of 20 L g -1 cm -1.
29.2 Use the Beer–Lambert relationship in quantita-
29.3 Determine the molar absorptivity of a substance
tive spectrophotometric analysis. Calculate the
in aqueous solution. A solution of p-nitrophenol
following (express your answer to 3 significant
containing 8.8 mg mL-1 gave an absorbance of
figures):
0.535 at 404 nm in a cuvette of path length 1 cm.
(a) The concentration (mg mL-1) of pentachloro- What is the molar absorptivity of a p-nitrophe-
phenol in a test solution giving an absorb- nol at 404 nm, expressed to 3 significant figures?
ance at 300 nm (A300) of 0.57 in a cuvette of (Note: Mr of p-nitrophenol is 291.27.)

30 Atomic spectroscopy
Atomic spectroscopy is a quantitative technique used for the determination of

All spectroscopic equipment is costly –
metals in samples. Atomic spectroscopy is characterised by two main tech-
The use of such equipment must always
niques: atomic absorption spectroscopy and atomic emission spectroscopy.
be done under guidance from a demon-
Atomic absorption spectroscopy (AAS) is normally carried out with a flame
strator or technician. All equipment in
(FAAS), although other devices can be used. Atomic emission spectroscopy
this section has an inherent risk due to
(AES) is typified by the use of a flame photometer (p. 222) or an inductively
its use of mains electricity.
coupled plasma. The flame photometer is normally used for elements in groups
I and II of the Periodic Table only, i.e. alkali and alkali earth metals.
In both AAS and AES, the substance to be analysed must be in solution.
Safety Note In atomic spectros- In order to do quantitative analysis, i.e. determine how much of the metal is
copy, the use of high-pressure gas present, the preparation of analytical standard solutions is necessary. While the
sources, e.g. cylinders, can be par- concentration range over which the technique can be used may be different, for
ticularly hazardous. Always consult a various instruments, the principles associated with the preparation of analytical
demonstrator or technician before use. standard solutions are the same (Boxes 30.1–30.5).
Atomic Absorption Spectroscopy

radiation
source The components of an atomic absorption spectrometer are a radiation source,
an atomisation cell, a sample introduction system, a method of wavelength
atomisation selection and a detector (Fig. 30.1).
cell
Radiation source
The main radiation source for AAS is the hollow-cathode lamp (HCL). The
wavelength
selection HCL (Fig. 30.2) emits radiation characteristic of a particular element. The
choice of HCL for AAS is simple. For example, if you are analysing for lead,
you will need a lead-coated HCL. It is normal to pre-warm the HCL for about
detection
and readout 10 min prior to use. This can be done either by using a separate pre-heater unit,
capable of warming up several HCLs simultaneously, or by inserting the HCL
Fig. 30.1 Components of an atomic absorp- in the AAS instrument and switching on the current. The lamp is typically
tion spectrometer. operated at an electric current between 2 and 30 mA.
Box 30.1 How to prepare a 1000 mg mL −1 stock solution of a metal ion from a metal salt
Stock solutions can be prepared directly from 3. Establish the ratio of Mr to Ar:
reagent-grade chemicals. It is important to use only 331.20
reagent-grade chemicals of the highest purity (e.g.
= 1.5985 g of Pb(NO3)2 in 1 litre
207.19
AnalaR®). This includes the water to be used – distilled 4. Accurately weigh out (p. 19) the metal salt. In this
and deionised – MilliQ water. (Note: many reagents (solids case, weigh 1.5985 g of Pb(NO3)2.
and liquids) contain metallic impurities in trace amounts.
While you can minimise this risk of contamination by 5. Quantitatively transfer the metal salt to a pre-cleaned
using the highest-purity reagents, it is essential to run 100 mL beaker and dissolve in 1% v/v HNO3 (AnalaR®
’reagent blanks’, especially for elemental determinations or equivalent).
trace levels.)
6. Quantitatively transfer the dissolved metal salt to a
1 L volumetric flask and make up to the graduation
1. Determine the Mr of the metal salt. For example, the mark with 1% v/v HNO3 (AnalaR® or equivalent).
Mr of Pb(NO3)2 = 331.20 g mol-1.
Often, a certified stock standard with a single or multi-
2. Determine the Ar of the metal. The Ar for Pb is element composition can be purchased, usually at a con-
207.19 g mol-1. centration (per element) of 1000 mg L-1 (1000 mg mL-1).

Atomic spectroscopy
Box 30.2 H
ow to prepare a set of five calibration solutions in the concentration range
0910 mg mL −1 (mg L −1)
Assuming that we are starting with a 1000 mg mL-1 stock 10 000 mg

K 100 mg mL-1 Pb
solution of a particular metal (e.g. lead) then you will need 100 mL
the following: six 100.00 mL grade A volumetric flasks; two
You now have a 100 mg mL-1 ’working’ stock solution
100 mL beakers; and a graduated pipette (0–10.00 mL).
of Pb.
1. Ensure that all the glassware is clean (see p. 68).
5. Transfer ≈15 mL of the working stock solution into
2. Transfer ≈15 mL of the stock solution into one of the the other pre-cleaned beaker.
pre-cleaned beakers.
6. Quantitatively transfer 2.00 mL of the solution into
3. Quantitatively transfer 10.00 mL of the stock solution a 100.00 mL volumetric flask and dilute to 100.00 mL
into a 100.00 mL volumetric flask. Then, dilute to with 1% v/v HNO3 (high purity). Label the flask as the
100.00 mL with 1% v/v HNO3 (high purity). 2 mg mL-1 Pb calibration solution.
4. Determine the concentration of this new solution. 7. Similarly transfer 0, 4.00, 6.00, 8.00 and 10.00 mL vol-
Remember that we started with an initial 1000 mg mL-1 umes into separate volumetric flasks and dilute to
Pb stock solution. 100.00 mL with the nitric acid and label as 0, 4, 6, 8
and 10 mg mL-1 Pb calibration solutions.
1000 mg
* 10 mL K 10 000 mg Pb
mL 8. Take the 0, 2, 4, 6, 8 and 10 mg mL−1 Pb calibration
10 000 mg Pb was placed in a 100.00 mL volumetric solutions for FAAS analysis.
flask, so:
Atomisation cell
Sample/standard dilutions – all dilu-
tions should be done using appropriate Several types of atomisation cell are available: flame, graphite furnace, hydride
glassware or plastic ware. Typically, this generation and cold vapour. Flame is the most common. In the pre-mixed lam-
involves the use of grade A pipettes for inar flame, the fuel and oxidant gases are mixed before they enter the burner
the transfer of known volumes of liq- (the ignition site) in an expansion chamber. The more commonly used flame
uids and grade A volumetric flasks for in FAAS is the air–acetylene flame (temperature, 2500 K), while the nitrous
subsequent dilutions. oxide-acetylene flame (temperature, 3150 K) is used for refractory elements
(e.g. Al). A complex series of processes occur in the flame resulting in the
formation of atoms. The aqueous sample solution, e.g. a metal salt, enters the
Safety Note Caution is needed flame in the form of an aerosol (Fig. 30.3). The flame is formed in a slot burner
when using strong (concentrated) positioned in the light path of the HCL (Fig. 30.4).
acids. When using concentrated acids In the graphite furnace atomiser, a small volume of sample (59100 mL)
always work in a fume cupboard. Wear is introduced onto the inner surface of a graphite tube (or onto a platform
gloves to protect your hands from ’acid placed within the tube) through a small opening (Fig. 30.5). The graphite tube
burns’. Always rinse affected areas with is arranged so that light from the HCL passes directly through the centre. Pass-
copious amounts of water. ing an electric current through the tube allows the operator to program a heating
cycle, with several stages (Fig. 30.6) including the elimination of water from
hollow cathode
the sample (drying), removal of the sample matrix (ashing), atomisation of the
connecting pins glass envelope
analyte (analysis), and removal of extraneous material (cleaning). An internal
silica window
gas flow of inert gas (N2 or Ar) during the drying and ashing stages removes
any extraneous material.
Hydride generation is a sample introduction technique exclusively for
elements that form volatile hydrides (e.g. As, Se, Sn). An acidified sample
solution is reacted with sodium borohydride solution, liberating the gaseous
anode Ne or Ar
hydride in a gas–liquid separator Fig. 30.7. The generated hydride is then
transported to an atomisation cell using a carrier gas. The atomisation cell is
Fig. 30.2 Components of a hollow-cathode normally an electrically heated or flame-heated quartz tube. Using arsenic as
lamp (HCL).

Atomic spectroscopy
Sample solution aerosol evaporates, resulting in

M+(aq) + X– (aq) = MX (s)
the formation of a solid aerosol
Solid particles melt and vaporise MX(s) = MX (g)
Resultant molecules or molecule aggregates MX(g) = M(g) + X (g)

decompose into individual atoms
Atoms may absorb energy, by collision, and

M(g) = M*(g) or M(g) = M+(g) + e–
become ions or excited atoms.
Atoms in the ground state absorb radiation

M(g) + hv = M*(g)
energy emitted by the hcl and are measured.
Fig. 30.3 Atom cell: flame processes
light path
of HCL
flame
location
sample injection port

expansion
chamber graphite tube
emission
from HCL
electrical heating
Fig. 30.4 Components of a slot burner for FAAS. Fig. 30.5 Schematic diagram of a graphite
furnace atomiser.
Sample + AsH3
reducing agent N2
To AAS
4
3
temperature
waste
2
5
1
time
Fig. 30.6 Heating cycle for a graphite furnace atomiser. 1. drying;

2. ashing; 3. analysis; 4. cleaning; 5. cooling Fig. 30.7 Gas-liquid separator.

Atomic spectroscopy
Box 30.3 How to analyse a sample using the method of standard additions in FAAS
The method of standard additions is used when the sam- 9. Determine the concentration of the metal in the
ple matrix may cause difficulties, for example chemical original sample. This can be done by taking into
interferences, in sample concentration determination. account the dilutions involved in the standard addi-
Standard additions allows any adverse effects to be over- tions method and any dilutions used to prepare the
come by incorporating a known amount of the sample in sample (see dilution factor, Box 30.5).
the calibration solutions.
1. Prepare a 1000 mg mL−1 stock solution (see Box 30.1). Table 30.1 Standard additions solutions
−1
2. Then, prepare a 100 mg mL working stock solution
Volumetric flask Volume of 100 mg Volume of
(see Box 30.2). (100.00 mL mL −1 working stock aqueous sample
3. You will also need to have prepared the sample. If the capacity) solution (mL) solution (mL)
sample is a solid you will need to digest the sample 1 0 10
(see Box 30.7).
2 1 10
4. An estimate of the metal concentration in the sam- 3 3 10
ple is required prior to carrying out standard addi-
tions so that the linear relationship between signal 4 5 10
(absorbance) and concentration is maintained. 5 7 10
5. You can then prepare the standard addition solu-

tions. This is most easily done as in Table 30.1.
1
6. Analyse the samples using FAAS. 0.8
absorbance
7. Plot the graph. The graphical output should appear 0.6

2.9 µg mL-1
as shown in Fig. 30.4. 0.4
0.2
8. The graph should contain several features: it must
0
have a linear response (signal against concentration); -4 -3 -2 -1 0 1 2 3 4 5 6 7
it does not pass through the origin; and extrapolation concentration/µg mL-1
of the graph is required until it intersects the x-axis
(e.g. 2.9 mg mL-1). Fig. 30.8 Standard additions graph.
Box 30.4 Sample size and certified reference materials
If a linear calibration graph (plot of concentration against powdered solid steel or alloy sample, or not 61.0 g for a
absorbance) has been prepared for lead in FAAS with the powdered biological sample such as citrus leaves). Often
concentrations 0, 2, 4, 6, 8 and 10 mg mL-1, the absorb- the maximum sample size is limited by the cost of the
ance for the digested sample needs to fall within the CRM. If using a ’real’ sample then it is best to take a larger
linear portion of the graph. This is controlled by two (non- sample size, since a CRM has usually been tested and
instrumental) factors: the weight of the sample digested prepared to a high specification with respect to drying,
and the final volume that the digested sample is made milling and shelf-life time. A typical minimum sample size
up to. As the volume of the digested sample is limited by for a soil might be 5.00 g. It is important if using ’real’
the availability of volumetric flasks (10.00, 25.00, 50.00, samples to consider the following additional factors:
100.00 or 250.00 mL are most commonly used, with the
●● Sampling – how it is to be done? How will a represent-
50.00 and 100.00 mL volumetric flasks the most common)
ative sample be arrived at?
it is often easier to alter the sample size. In order to have
a representative sample, a minimum sample size is often ●● Storage of sample – what containers will be used for
recommended. For example, if using a certified reference storage of sample? Be aware of contamination for the
material (CRM) the supplier will recommend a minimum storage container and from the implements used to
sample size to ensure homogeneity (e.g. not 60.5 g for a sample and transfer the sample.
(continued)

Atomic spectroscopy
●● Lifetime of stored sample – how long will the sam- concentration of various metals within the sample as well
ple remain stable? Is preservation of the sample as their variation, normally quoted as one standard devi-
necessary? ation either side of the mean value (e.g. 2.5 { 0.3 mg g -1
Pb). CRMs are used to test the accuracy of a new method
Note: CRMs can be obtained from appropriate suppliers,
or to enable a quality control scheme to be operated by a
for example the Laboratory of the Government Chemist
commercial laboratory (see also Chapter 47). In practical
(LGC) in the UK or the National Institute for Science and
work, they are useful for assessing student performance
Technology (NIST) in the USA. In addition to the CRM, a
in preparing and analysing a sample.
certificate is provided that contains information on the
Box 30.5 Analysis of a sample: dilution factor
A sample was weighed (0.4998 g) and digested in con- 3.4 mg 100 mL

centrated nitric acid (20 mL). After cooling, the digested * = 680 mg g -1
mL 0.4998 g
sample was quantitatively transferred into a 100.00 mL
volumetric flask and made up with ultrapure water and Note:
then analysed for lead by FAAS. Let us suppose that the
●● The volume of acid used is irrelevant in the
absorbance obtained corresponds to a concentration of
calculation – only the final volume in the volumetric
3.4 mg mL-1. What is the concentration of lead in the orig-
flask matters.
inal sample?
The method of calculation is most appropriately done ●● The units cancel (mL on top line cancels with mL on the
as follows: bottom line) leaving you with units of mg g -1 (mg/g).
●● Calculate the dilution factor. This can be done if the ●● Alternatively, the units can be expressed in mg kg-1
final volume of the sample and its original weight are (mg/kg), i.e. 680 mg kg-1 or % w/w, i.e. 0.068% w/w
known. In this case 100 mL and 0.4998 g. (see p. 103). (10 000 mg g -1 K 1% w/w or for aqueous
samples 10 000 mg mL-1 K 1% w/v.)
●● You then multiply the concentration from the graph
with the dilution factor:
an example it is possible to write the following equation for the generation

of arsine (AsH3):
3BH4- + 3H + + 4H3AsO3 S 3H3BO3 + 4AsH3 + 3H2O[30.1]
Cold vapour generation is the term exclusively reserved for mercury. Mercury
in a sample is reduced to elemental mercury by tin (II) chloride (eqn 30.2):
Sn2+ + Hg 2+ S Sn4+ + Hg 0[30.2]
and the mercury vapour produced is transported to an atomisation cell by a

carrier gas. The atomisation cell consists of a long-path glass absorption cell
Safety Note Once the flame is located in the path of the HCL (Fig. 30.9). Mercury is monitored at a wave-
ignited the instrument should not be length of 253.7 nm.
left unattended. Sample introduction into the flame
Samples are almost exclusively introduced into flames as liquids. Solid samples
Safety Note Check that the drain need to be converted to aqueous solutions using methods such as decomposi-
is full of water prior to use.
tion (see p. 238). Once in the aqueous form the sample is introduced into the
flame using a nebuliser–expansion chamber (Fig. 30.10).

Atomic spectroscopy
hcl
silica tube The pneumatic concentric nebuliser (see also p. 234) consists of a stain-
N2
less steel tube through which a Pt/Ir capillary tube is located (Fig. 30.10). The
Sample + SnCl2 aqueous sample is drawn up through the capillary tube by the action of the
oxidant gas (air) escaping through the exit orifice that exists between the out-
Hg(O) side of the capillary tube and the inside of the stainless steel tube. The action
of the escaping air and aqueous sample is sufficient to form a coarse aerosol in
a process termed the Venturi effect. The typical uptake rate of the nebuliser is
waste between 3 and 6 mL min -1.
The expansion chamber (Fig. 30.10) has two functions. The first is con-
cerned with aerosol generation, the objectives of which are:
●● to convert the aqueous sample solution into a coarse aerosol using the oxi-
diagram of cold vapour AAS dant gas;
Fig. 30.9 Diagram of cold vapour AAS. ●● to disperse the coarse aerosol further into a fine aerosol, by interaction with
baffles located within the chamber;
fine aerosol ●● to condense any residual aerosol particles, which then go to waste.
to burner
fuel coarse
aerosol The second function involves the safe pre-mixing of the oxidant and fuel gases
Pt/Ir capillary before they are introduced into the laminar flow burner.
expansion
or cloud Wavelength selection and detection
sample chamber
As AAS is used to monitor one metal at a time, the spectrometer used is termed
oxidant a monochromator. Two optical arrangements are possible; single and double
baffles beam. The latter is preferred as it corrects for fluctuations in the HCL caused
water trap
by warm-up, drift and source noise, thus leading to improved precision in the
absorbance measurement. A schematic diagram of the optical arrangement
Fig. 30.10 Schematic diagram of a nebuliser– is shown in Fig. 30.11. The attenuation of the HCL radiation by the atomic
expansion chamber for FAAS. vapour is detected by a photomultiplier tube (PMT), a device for proportionally
converting photons of light to electric current. An illustration of the overall
principle of AAS is summarised in Fig. 30.12.
Emission from hcl Element-specific

spectrum
Absorption in the flame
Czerny–Turner The decrease in peak height is

monochromator proportional to the atom’s
concentration in the flame
grating
photomultiplier
atom tube Monochromator
HCL cell
PC readout
Readout
chopper
half-silvered mirror
Spectral slit width
Fig. 30.11 Schematic diagram of the optical
arrangement for AAS. Fig. 30.12 The principle of AAS.

Atomic spectroscopy
Background correction methods

One of the main practical problems with the use of AAS is the occurrence of
molecular species that coincide with the atomic signal. One approach to remove
this molecular absorbance is by the use of background correction methods.
Several approaches are possible, but the most common is based on the use
of a continuum source, D2. In the atomisation cell (e.g. flame), absorption is
possible from both atomic species and from molecular species (unwanted inter-
ference). By measuring the absorption that occurs from the radiation source
(HCL) and comparing it with the absorbance that occurs from the continuum
source (D2) a corrected absorption signal can be obtained. This is because the
atomic species of interest absorb the specific radiation associated with the HCL
source, whereas the absorption of radiation by the continuum source for the
same atomic species will be negligible.
Interferences in the flame

Interferences in the flame can be classified into four categories: chemical, ion-
isation, physical and spectral.
Chemical interferences occur when the analyte forms a thermally stable
compound with a molecular or ionic species present in the sample solution.
Flame interferences Examples include the suppression of alkaline earth metals due to the presence
of phosphate, silicate or aluminate in the sample solution in the air–acetylene
●● chemical flame. The best known example of this is the absorption signal suppression that
●● ionisation
occurs for Ca at 422.7 nm owing to increasing amounts of phosphate. This sig-
●● physical
nal suppression is due to the formation of calcium pyrophosphate, a thermally
●● spectral
stable compound in the flame.
Ionisation interferences occur most commonly for alkali and alkaline earth
metals. The low ionisation potential of these metals can lead to their ionisation
in the relatively hot environment of the flame. If this occurs, no absorption sig-
nal is detected, since FAAS is a technique for measuring atoms not ions. This
process can be prevented by the addition of an ionisation suppressor or ‘buffer’
(e.g. an alkali metal such as Cs). Addition of excess Cs leads to its ionisation in
The resonance wavelength results the flame in preference to the metal of interest (e.g. Na). This process is termed
from an energy transition from a higher the ‘mass action’ effect.
energy level to the ground state. In Physical interferences are due to the effects of the sample solution on
practice the highest absorbance signals aerosol formation within the spray chamber. The formation of an aerosol is
are obtained when using the resonance dependent upon the surface tension, density and viscosity of the sample solu-
wavelengths. tion. This type of interference can be controlled by the matrix matching of
sample and standard solutions, i.e. add the same sample components to the
standard solution, but without the metal of interest. If this is not possible, it is
then necessary to use the method of standard additions (Box 30.3).
Spectral interferences are uncommon in AAS owing to the selectivity of
Table 30.2 Examples of spectral interferences the technique Table 30.2. However, some interferences may occur, for exam-
ple the resonance line of Cu occurs at 324.754 nm and has a line coincidence
Resonance
from Eu at 324.753 nm. Unless the Eu is 1000 times in excess, however,
wavelength Wavelength
Analyte (nm) Interferent (nm) it is unlikely to cause any problems for Cu determination. In addition to
atomic spectral overlap, molecular band absorption can cause problems, for
Cu 324.754 Eu 324.753 example calcium hydroxide has an absorption band on the Ba wavelength
Fe 271.902 Pt 271.904 of 553.55 nm while Pb at 217.0 nm has molecular absorption from NaCl.
Al 308.215 V 308.211 Molecular band absorption can be corrected for using background correc-
tion techniques. The operation of a flame atomic absorption spectrometer is
Hg 253.652 Co 253.649

Atomic spectroscopy
Box 30.6 How to operate a flame atomic absorption spectrometer
You should only operate an FAAS system under direct 9. Press the ignite button (or flame button). The flame
supervision. The instrument should be located under a should light instantaneously with a ’pop’.
fume extraction hood. The spectrometer requires approxi-
mately 20 min. to warm up before switching the gases on 10. After establishing the flame, insert the aspirator
and using the instrument. tube into distilled water. Allow the flame to stabilise
for up to 1 minute by aspirating distilled water, prior
1. Adjust the operating wavelength and slit width of the to analysis.
monochromator. This is done by consulting stand-
ard operating conditions, for example for lead see 11. After completing your analysis, shut off the acety-
Table 30.3. lene first (by closing the cylinder valve) and vent the
acetylene gas line while the air is still on. Then, shut
2. Decide what wavelength is to be used for the off the air compressor and allow the air line to vent.
a nalysis. For lead the maximum sensitivity is
achieved by selecting 217.0 nm. 12. Finally, switch off the fume extraction hood.
3. Adjust the wavelength selector to the appropriate

wavelength.
Table 30.3 Standard operating conditions for lead
4. Adjust the gain control until the energy meter read-
ing reaches a maximum. Wavelength Slit Characteristic concentration
(nm) (nm) (mg L −1)
5. Adjust the wavelength selector for maximum signal
reading. You are now ready to ignite the air–acetylene 283.3 0.7 0.45
flame.
217.0 0.7 0.19
6. Turn on the fume extraction hood. This allows toxic 205.3 0.7 5.4
gases to be safely removed from the laboratory
environment. 202.2 0.7 7.1
261.4 0.7 11.0
7. Turn on the air supply such that the oxidant flow
meter is at the desired setting. 368.3 0.7 27.0
364.0 0.7 67.0
8. Turn on the acetylene supply such that the fuel flow
meter is at the desired setting. Note: recommended flame: air-acetylene, oxidising (lean, blue).
Atomic emission spectroscopy

The main components of an atomic emission spectrometer are an atomisation
and ionisation cell, a method of sample introduction, the spectrometer and
detector. In contrast to AAS, no radiation source is required.
Flame photometry (see also p. 222) is almost exclusively used for the
determination of alkali metals because of their low excitation potential (e.g.
Using a flame photometer – make sure sodium 5.14 eV and potassium 4.34 eV). This simplifies the instrumentation
distilled water is available to aspirate required and allows a cooler flame (air–propane, air–butane or air–natural
into the flame once it is ignited. gas) to be used in conjunction with a simpler spectrometer (interference filter).
The use of an interference filter allows a large excess of light to be viewed
by the detector. Thus, the expensive photomultiplier tube is not required and
a cheaper detector can be used, for example a photodiode or photoemissive
detector. The sample is introduced using a pneumatic nebuliser as described
Contamination risk – wipe the outside for FAAS (p. 231). Flame photometry is therefore a simple, robust and inex-
of the aspirator tube with a clean tis- pensive technique for the determination of potassium (766.5 nm) or sodium
sue in between samples/standards to (589.0 nm) in clinical or environmental samples. The technique suffers from
prevent contamination. the same type of interferences as in FAAS. The operation of a flame photom-
eter is described on p. 223.

Atomic spectroscopy
Inductively coupled plasma

A radio frequency inductively coupled plasma (ICP) is formed within the con-
fines of three concentric glass tubes or plasma torch (Fig. 30.13). Each concen-
tric glass tube has a tangentially arranged entry point through which argon gas
enters the intermediate (plasma) and external (coolant) tubes. The inner tube con-
sists of a capillary tube through which the aerosol is introduced from the sample
Be prepared – ensure all standards, introduction system. Located around the plasma torch is a coil of water-cooled
blanks and samples are readily and eas- copper tubing. Power input to the ICP is achieved through this copper, load or
ily accessible prior to ignition. induction coil, typically in the range 0.5–1.5 kW at a frequency of 27 or 40 MHz.
Initiation of the plasma is achieved as follows. The carrier gas flow is
first switched off and a spark added momentarily from a Tesla coil (attached
to the outer edge of the plasma torch). The spark, a source of ‘seed’ electrons,
causes ionisation of the argon gas. The co-existence of argon, argon ions and
electrons constitutes a plasma located within the confines of the plasma torch
but protruding from the top in the shape of a bright white luminous bullet. In
order to introduce the sample aerosol into the ICP (7000–10 000 K) the carrier
gas is switched on and punches a hole into the centre of the plasma creating
the characteristic doughnut or toroidal shape. The emitted radiation is viewed
laterally (side-on) through the luminous plasma.
Sample introduction
load coil The most common method of liquid sample introduction in ICP–AES is the
nebuliser (Fig. 30.14). The nebuliser operates in the same manner as that used
for FAAS but there are differences in its construction material and manufac-
injector tube
tured tolerance (the nebuliser for ICP–AES generates a finer aerosol, but is
more inefficient). The pneumatic nebuliser consists of a concentric glass tube
through which a capillary tube passes (Fig. 30.14(a)). The sample is drawn up
through the capillary by the action of the argon carrier gas escaping through the
exit orifice that exists between the outside of the capillary tube and the inside of
the glass concentric tube. The typical uptake rate of the nebuliser is between 0.5
intermediate gas flow and 4 mL min -1. In common with FAAS, a means to reduce the coarse aerosol
generated to a fine aerosol is required. In ICP–AES terminology this device is
outer gas flow called a spray chamber (Fig. 30.15).
(a) Concentric (b) Cross-flow (c) High-solids
sample aerosol Sample

(pumped)
Fig. 30.13 Schematic diagram of an induc- Argon gas
tively coupled plasma. V-groove
Fig. 30.14 Types of nebulisers.
Double-pass Cyclonic Impact-bead

ICP
Impact bead
Side view Top view
Nebuliser Nebuliser
Nebuliser
Waste
Waste
Waste
Fig. 30.15 A range of nebuliser-spray chamber combinations.

Atomic spectroscopy
Lens
Mirror Spectrometers
Source The nature of the ICP is such that all elemental information from the sample
is contained within it. The only limitation is whether it is possible to observe
all the elemental information at the same time or one element at once. This
limitation is associated not with the ICP but with the type of spectrometer used
Grating
Mirror to view the emitted radiation. A monochromator allows measurement of one
Prism wavelength, corresponding to one element at a time, while a polychromator
allows multiwavelength or multielement detection. The former can perform
Detector sequential multielement analysis, while the latter carries out simultaneous
multielement analysis. The typical wavelength coverage required for a spec-
trometer is between 167 nm (Al) and 852 nm (Cs). A schematic diagram of a
Fig. 30.16 Spectral diagram of the optical modern Echelle spectrometer is shown in Fig. 30.16.
layout of an Echelle spectromater
Detectors
240 200
The most common detector for AES is the photomultiplier tube (see p. 231).
220
An alternative approach for the detection of multielement (multiwavelength)
260 240 220
information is the charged-coupled device (CCD). A CCD is essentially an
300 280 260
array of closely spaced metal–insulator–semiconductor diodes formed on a
Prism dispersion
280
340 320 300 wafer of semiconductor material. Incident light striking the CCD is converted
360
order
400 380 340 320 into an electrical signal. A typical spectral map output is shown in Fig. 30.17.
500 480 460 440 420
620 600 580 560 540 520 500 Interferences in ICP–AES
800 780 760 740 720 700 680 660 640 620 Interferences for AES can be classified into two main categories, spectral and
matrix interferences. Spectral interference can occur as a result of an interfer-
Wavelength (nm)
ing emission line from either another element or the argon source gas, or from
Grating dispersion impurities within or entrained into the source, for example, molecular species
such as N2. Such interferences can be eliminated or reduced either by increas-
Fig. 30.17 Spectral map generated by the ing the resolution of the spectrometer or by selecting an alternative spectral
Echelle spectrometer. emission line.
Matrix interferences are often associated with the sample introduction pro-
cess. For example, pneumatic nebulisation can be affected by the dissolved-
Sample cone Skimmer cone solids content of the aqueous sample, which affects the uptake rate of the
nebuliser and hence the sensitivity of the assay. Matrix effects in the plasma
Tesla coil source typically involve the presence of easily ionisable elements (EIEs, e.g.
alkali metals) within the plasma source.
Inductively coupled plasma mass spectrometry

The inductively coupled plasma has been developed as an ion source for mass
spectrometry (see also Chapter 38). The plasma is operated in horizontal mode
and is positioned at the interface of a mass spectrometer. Most common mass
spectrometers for ICP–MS are based on a quadrupole instrument. Key to the
ICP plasma success of ICP–MS was the development of an interface (Fig. 30.18) that allows
torch
an ICP operating at atmospheric pressure to be linked to a mass spectrometer
operating under high vacuum.
The ICP operates in much the same way as in AES, and with the same
approach to sample introduction. However, the inherent sensitivity of ICP–MS
Atmospheric moderate High (Fig. 30.19) has made it a very useful addition to the methods available for
pressure vacuum vacuum
trace metal analysis. Typical sensitivities for metals by ICP–MS are in the trace
Fig. 30.18 Schematic diagram of an induc- (ng ml-1) to ultratrace (pg mL-1) range. This ability to measure metals at low
tively coupled plasma mass spectrometer concentrations can cause problems, particularly with the grade of reagents used
interface. to prepare the standards and the water supply to prepare samples and standards.

Atomic spectroscopy
Mass
Interface Lenses spectrometer Detector
ICP
Data
output
Vacuum Vacuum Vacuum

pump pump pump
Fig. 30.19 Schematic diagram of an inductively coupled plasma mass

spectrometry system.
Quantitative analysis in ICP–MS is achieved by measurement of element

isotopes. Approximately 70% of elements in the Periodic Table have more than
one stable isotope. For example, lead (Pb) has four isotopes with mass/charge
ratios of 204, 206, 207 and 208 which have % abundances of 1.4, 24.1, 22.1
and 52.4, respectively. It should therefore be obvious that in order to achieve
the largest response for Pb you should choose the isotope with the largest
abundance, i.e. 208. However, a mass spectrometer offers additional advan-
tages, including the ability to measure all isotopes very quickly. An ICP–MS
can therefore measure almost all elements in the Periodic Table very quickly.
ICP–MS can also use the isotopic information present for an element to provide
quantitative information. The typical approach to quantitation involves pre-
paring a calibration graph (see Chapter 48). However, ICP–MS can also quan-
tify samples using isotope dilution analysis (IDA). Isotope dilution analysis
requires an enriched artificial isotopic standard of the element to be quantified,
i.e. a different isotopic abundance to that normally found in nature.
Two classes of interferences occur in ICP–MS:
1. isobaric interferences;
2. molecular interferences (polyatomic and doubly charged species).
Isobaric interferences occur as a result of direct overlap of isotopes from
one element with another. These interferences (Table 30.4) are well known
and can be compensated for either by selecting an alternative isotope or by
applying a correction based on the isotopic abundance. Molecular interfer-
ences, and in particular polyatomic interferences, can be troublesome and
result from interactions between the element of interest and other species
present, for example argon plasma gas, or the type of acid used to prepare
the sample digest. It is therefore possible to produce species, such as ArCl+
or, more correctly in this example, 40Ar35Cl+ . This may have resulted from
the presence of chlorine in the sample matrix or the acid used to digest the
sample (e.g. HCl).
The other molecular interference, doubly charged species, can also cause
problems. For example, barium has isotopes at mass/charge ratios that include
130, 132, 134, 136 and 138 amu that can form doubly charged species at mass/
charge ratios of 65, 66, 67, 68 and 69 amu, respectively.

Atomic spectroscopy
Table 30.4 Examples of isobaric interferences
Element Atomic mass unit (amu)
108 109 110 111 112 113 114 115 116 117 118 119 120
Pd 26.5 11.7
Ag 48.2
Cd 0.9 12.5 12.8 24.1 12.2 28.7 7.5
In 4.3 95.7
Sn 1.0 0.6 0.4 14.5 7.7 24.2 8.6 32.6
Te 0.09
One approach to reduce or eliminate molecular interferences is via a

c ollision–reaction cell (Dean, 2005). The use of a collision–reaction cell can
allow for either neutralisation of chemical ionisation species or interferent/ana-
lyte ion mass/charge ratio shifts. These charges are affected by the introduction
of reagent gases (e.g. H2, He, NH3 or CH4) into the collision–reaction cell.
Reaction types involved in a collision-reaction cell include:
●● charge exchange;
●● atom transfer;
●● adduct formation;
●● condensation reactions.
Charge exchange Charge exchange allows the removal of, for example, the
argon plasma gas ion interference and the resultant formation of uncharged
argon plasma gas. which is not then detected,
Ar + + NH3 S NH3+ + Ar[30.3]
Atom transfer: proton transfer Proton transfer can remove the interference
from, for example, ArH + . This results in the formation of neutral (uncharged)
argon plasma gas which is then not detected.
ArH + + H2 S H3+ + Ar[30.4]
Atom transfer: hydrogen-atom transfer Hydrogen-atom transfer lias the

ability to alleviate an interference by incresing the mass/charge ratio by one.
Ar + + H2 S ArH + + H[30.5]
Adduct formation Adduct formation, with, for example, ammonia. NH3,

allows the mass/charge ratio to increase by 17 amu (atomic weight of N = 14
Example isobaric interference (Table
and atomic weight of H = 1).
30.4) – If analysing for Cd the most
abundant isotope is 114Cd (with an
Ni+ + NH3 S Ni+ ~ NH3[30.6]
abundance of 28.7%). However, 114amu
also coincides with a minor abundant
Condensation reaction The use of a condensation reaction, in common with
isotope (0.6%) for 114Sn.
adduct formation, has the ability to increase the mass/charge ratio. For example,

Atomic spectroscopy
creation of the oxide of the element will increase the mass/charge ratio by
16 amu (atomic weight of O = 16).
Ce + + N2O S CeO+ + N2[30.7]
Decomposition techniques for solid inorganic samples

Conversion of a solid matrix into a liquid matrix involves the decomposition
of the sample. One of the major problems in preparing solid samples for trace
element analysis is the potential risk of contamination. Contamination can arise
from several sources: the grade of reagents used; the vessels used for digestion
and the subsequent dilution of the sample; and human involvement.
In order to minimise the risk of contamination you should take the fol-
lowing measures:
●● Use the highest purity of reagents and acids, including the water used for
sample dilution.
●● Use sample blanks in the analytical procedure, to identify the base level of
impurity in the reagents.
●● Soak sample vessels in an acid leaching bath (e.g. 10% v/v nitric acid) for
at least 24 hours, followed by rinsing in copious amounts of ultrapure water.
●● Store cleaned volumetric flasks with their stoppers inserted; cover beakers
with Clingfilm® or store upside down to protect from dust.
●● In addition to the wearing of a laboratory coat and safety glasses, it may be
necessary to wear ‘contaminant’-free gloves and a close-fitting hat.
Decomposition involves the liberation of the analyte (metal) of interest from
an interfering matrix using a reagent (mineral/oxidising acids or fusion flux)
and/or heat. An important aspect in the decomposition of an unknown sample
is the sample size (Box 30.4). You need to consider two aspects. Firstly, the
dilution factor required to convert the solid sample to an aqueous solution
(Box 30.5), and, secondly, the sensitivity of the analytical instrument (e.g.
FAAS or ICP-MS).
Acid digestion
This involves the use of mineral or oxidising acids and an external heat source
to decompose the sample matrix. The choice of an individual acid or combi-
nation of acids depends upon the nature of the matrix to be decomposed. For
example, the digestion of a matrix containing silica, SiO2 (e.g. a geological
sample), requires the use of hydrofluoric acid (HF). A summary of the most
common acids used for digestion and their application is shown in Table 30.5.
Once you have chosen an appropriate acid, place your sample into an
appropriate vessel for the decomposition stage. Typical vessels include an open
glass beaker or boiling tube for conventional heating (Fig. 30.20) or for micro-
wave heating, a PTFE or Teflon® PFA (perfluoroalkoxyvinylether) vessel. A
typical microwave system (Fig. 30.21) operates at 2.45 GHz with up to 14
sample vessels arranged on a rotating carousel; commercial systems have addi-
tional features such as a PTFE-lined cavity; a safety vent (if the pressure inside
a vessel is excessive the vent will open, allowing the contents to go to waste);
and an ability to measure both the temperature and pressure inside the digestion
vessels. A procedure for acid digestion of a sample is shown in Box 30.7.

Atomic spectroscopy
Table 30.5 Common acids* used for digestion
Acid(s) Boiling point (°C) Comments
Hydrochloric acid (HCl) 110 Useful for salts of carbonates, some oxides and some sulphides. A weak
reducing agent; not generally used to dissolve organic matter
Hydrofluoric acid (HF) 112 For digestion of silica-based materials only. Cannot be used with glass
containers (use plasticware). In addition to laboratory coat and safety glasses,
extra safety precautions are needed, e.g. gloves. In case of spillages, calcium
gluconate gel is required for treatment of skin contact sites and should be
available during use; evacuate to hospital immediately if skin is exposed
to liquid HF
Nitric acid (HNO3) 122 Useful for the digestion of metals, alloys and biological samples. Oxidising
attack on many samples not dissolved by HCl; liberates trace metals as the
soluble nitrate salt
Sulphuric acid (H2SO4) 338 Useful for releasing a volatile product; good oxidising properties for ores,
metals, alloys, oxides and hydroxides. Often used in combination with
HNO3. Note: Sulphuric acid must never be used in PTFE vessels (melting
point 327 °C)
Hydrochloric/nitric acids – A 3 : 1 v/v mixture of HCl and HNO3 is called aqua regia. It forms a reac-
(HCl/HNO3) tive intermediate, NOCl. Useful for digesting metals, alloys, sulphides and
other ores
* All concentrated acids should be used only in a fume cupboard.
Heating block
Sample tube
Sample well
Acid
Sample
Fig. 30.20 Schematic of a commercial acid-digestion system.
Temperature and
pressure sensor
connectors Wave guide Magnetron antenna
Magnetron
Mode
Isolated electronics
Cavity stirrer
exhausted
To chemical Room air inlet
fume hood
Chemically resistant
coating on cavity walls
Fig. 30.21 Schematic diagram of a commercial microwave digestion system.

Atomic spectroscopy
Box 30.7 How to acid-digest a sample using a hot plate
1. Accurately weigh your sample into a beaker (100 mL). 7. Wash the watch-glass cover into the beaker to ’cap-
For digestion of a powdered metal sample 0.5000 g is ture’ any splashes of solution.
appropriate (for details on how to weigh accurately
see p. 74). 8. Dilute the digested sample with deionised, distilled
water.
2. Add 20 mL of concentrated acid(s) (see Table 30.5).
9. Quantitatively transfer the diluted, digested sample
3. Cover the beaker with a watch glass. This is done to to a 100.00 mL volumetric flask (see p. 75). Make
prevent the loss of sample and to minimise the risk of up to the graduation mark with de-ionised, distilled
contamination. water.
4. Place the beaker on a pre-heated hot plate. 10. Prepare a sample blank using the same procedure,
5. Reflux the sample for approx. 30 min to 1 hour; i.e. perform all of the above tasks but without adding
depending on the nature of the sample a coloured, the actual sample.
clear solution should result.
11. Prepare samples in at least duplicate. For statistical
6. Remove the beaker from the heat and allow to cool. work on the results, at least seven sample digests and
This may take several minutes. Retain the watch- two sample blanks are recommended.
glass cover during this stage to reduce airborne
contamination.
Other methods of sample decomposition

The use of acid(s) and heat is probably the most common approach to the
decomposition of samples. However, several alternatives exist, including dry
ashing and fusion.
Dry ashing involves heating the sample in air in a muffle furnace
(Fig. 30.22) at 400–800 °C to destroy the sample matrix (e.g. soil). After
decomposition, the sample residue is dissolved in acid and quantitatively trans-
ferred to a volumetric flask prior to analysis. The method may lead to the loss
of volatile elements (e.g. Hg, As).
Some substances, such as silicates and oxides, are not always destroyed by
the direct action of acid and heat. In these situations an alternative approach is
Fig. 30.22 A muffle furnace.
required. Fusion involves the addition of a ten-fold excess of a suitable reagent
(e.g. lithium metaborate or tetraborate) to a finely ground sample. The mixture
is placed in a metal crucible (e.g. Pt), and then heated in a muffle furnace at
900–1200 °C (Fig. 30.23). After heating (from several minutes to several hours)
a clear ‘melt’ should result, indicating completeness of the decomposition.
After cooling, the melt is dissolved in HF (Table 30.5). This process can lead
to a higher risk of contamination.
Fig. 30.23 Fusion.

Atomic spectroscopy
Text references and sources for further study

Cullen, M. (2003) Atomic Spectroscopy in Elemental Anal- Kellner, R., Mermet, J.M., Otto, M., Valcarcel, M. and
ysis. John Wiley & Sons Ltd, Chichester. Widmer, H.M. (2004) Analytical Chemistry: A Modern
Approach to Analytical Science, 2nd edn. John Wiley &
Dean, J.R. (1997) Atomic Absorption and Plasma Spec-
troscopy, 2nd edn. ACOL series, John Wiley & Sons
Ltd, Chichester. Lajunen, L.H.J. and Peramaki, P. (2004) Spectrochemical
Analysis by Atomic Absorption and Emission, 2nd edn.
Dean, J.R. (2005) Practical Inductively Coupled Plasma Royal Society of Chemistry, Cambridge.
Spectroscopy. John Wiley & Sons Ltd, Chichester.
Schmidt, W. (2005) Optical Spectroscopy in Chemistry and
Ebdon, L., Evans, H., Fisher, A. and Hill, S. (1998) An Life Sciences: An Introduction. John Wiley & Sons Ltd,
Introduction to Atomic Absorption Spectrometry. John Chichester.
Wiley & Sons Ltd, Chichester. Taylor, H. (2000) Inductively Coupled Plasma–Mass Spec-
Hill, S.J. (2007) Inductively Coupled Plasma Spectrometry trometry: Practices and Techniques. Academic Press,
and its Applications, 2nd edn. John Wiley & Sons Ltd, London.
Chichester. Vandecasteele, C. and Block, C.B. (1997) Modern Methods
Holland, J.G. and Tanner, S.D. (2003) Plasma Source Mass of Trace Element Determination. John Wiley & Sons
Spectrometry: Applications and Emerging Technologies. Ltd, Chichester.
Royal Society of Chemistry, Cambridge.
Study exercises
30.1 Determine the concentration of metal ions based Express your answer in mmol K + (g sample) -1, to
on atomic spectroscopy of test and standard 3 significant figures.
solutions. The following data represent a set of
calibration standards for K + in aqueous solution, 30.2 Determine unknowns from a calibration
measured by flame photometry: curve. The following data are for a set of cali-
bration standards of Zn, measured by atomic
Absorbance of standard solutions containing absorption spectroscopy.
K + at up to 0.5 mmol L-1
Absorbance measurements for a series of
Absorbance standard solutions containing different amounts
K+ concentration (mmol L −1)
of zinc:
0 0.000
0.1 0.155 Zinc concentration (mg mL -1) Absorbance
0.2 0.279
0.3 0.391 0 0.000
0.4 0.537 1 0.082
0.5 0.683 2 0.174
3 0.257
4 0.340
Draw a calibration curve using the above data
5 0.408
and use this to estimate the amount of K + in a 6 0.463
test sample prepared by digestion of 0.482 g of 7 0.511
sample in a final volume of 25 ml of solution, 8 0.543
giving an absorbance of 0.429 when measured 9 0.561
10 0.575
at the same time as the standards shown above.

Atomic spectroscopy
Draw a calibration curve by hand using graph (b) a 20-fold dilution, giving an absorbance of
paper and estimate the concentration of zinc in 0.304;
the following water samples: (c) a five-fold dilution, giving an absorbance of
0.550.
(a) an undiluted sample, giving an absorbance
of 0.157; Give your answer to three significant figures in
each case.

31 X-ray fluorescence spectroscopy
X-ray fluorescence spectrometry (XRF) can be applied to determine the

Terminology in XRF – It is common
elemental composition of a wide range of objects of importance in chem-
practice to refer to an atom with shells
ical sciences. Its value arises from its non-destructive nature and potential
(Fig. 31.1) of electrons, i.e. K, L, M, N,
for rapid identification of elements present in samples. The multi-element
etc., instead of the modern approach
nature of XRF allows ‘elemental fingerprints’ to be generated from samples
using s, p, d and f as descriptors.
of known or unknown authenticity. Typical applications areas include paint,
glass, pottery and ceramics, metals and alloys, soil, plastics, and fabrics. XRF
can also be used to identify elements present in drug samples that may give
clues as to the manufacturing processes involved and thereby help to further
characterise the samples.
Understanding the relationship between X-rays are part of the electromagnetic spectrum (p. 217) with wavelengths
energy and wavelength – The following between 0.001 and 10 nm. They are produced when high-energy electrons
equation applies: decelerate or when electron transitions occur in the inner shells of atoms. The
irradiation of matter with high-energy photons can lead to the ejection of an
E = h * c/l
electron from an inner shell (e.g. the K shell) of an atom (Fig. 31.1a) and
where E is the energy, h is Planck’s conformation of an ion. This vacancy in the inner shell is filled almost immedi-
stant, c is the velocity of light and l is ately by an electron from a higher energy level (Fig. 31.1b). The difference
the wavelength. in energy between the two energy levels is released in the form of an X-ray.
This energy difference between the two specific orbitals always has the same
characteristic energy. Therefore, by determining the energy (or wavelength)
of the X-ray emitted by a particular element, it is possible to determine the
Ejected identity of that element. In terms of identification, if the transition is to a K
electron
shell, the X-ray produced is described as a K X-ray, if to an L shell, an L X-ray
results, and so on. Each X-ray can be further classified as, for example, Ka, Kb,
indicating that the energy transition has taken place from the L and M shell
High speed
photon respectively (Fig. 31.2). The number of X-rays per unit time at a particular
energy (or wavelength) can be counted to allow either qualitative or quantitative
analysis to be undertaken.
Key Point The energy of an emitted X-ray is specific to a par-

ticular element and allows unequivocal identification while the
Nucleus
Electron
strength of the signal allows quantification.
Electron
shell
Electron drops
from outer shell
Instrumentation
An XRF spectrometer requires a source of X-rays (to excite the atoms in the
sample), a sample holder, and a spectrometer to measure the energy (or wave-
length) and intensity of the radiation emitted by the sample. Instrumentally two
types of XRF can be identified: energy-dispersive and wavelength-dispersive
XRF. Energy dispersive X-ray fluorescence (EDXRF) relies on the detector
and associated electronics to resolve spectral peaks due to differences in the
energy of the generated X-rays. A schematic diagram of an EDXRF is shown
in Fig. 31.3. In wavelength dispersive XRF (WDXRF) a diffractive device such
X-ray as a crystal is used to investigate the element of interest. A schematic diagram
emitted
of a WDXRF is shown in Fig. 31.4. This chapter will focus on EDXRF only,
Fig. 31.1 Operating principle of X-ray flu- since this is the most widely used method in analytical science. The analytical
orescence spectroscopy. Simplified dia- performance of EDXRF can be improved by the introduction of additional
grams showing (a) ejection of an electron components, including source filters or secondary targets. In the case of a sec-
and (b) the X-ray emitted. ondary target, the X-ray tube excites the secondary target. The secondary target

X-ray fluorescence spectroscopy
fluoresces and excites the sample, and finally the detector measures X-rays
from the sample. In addition, detector filters are sometimes positioned between
the sample and the detector to remove unwanted X-rays.
Sample chambers in XRF can be operated in air. However, as air absorbs
Kb La low-energy X-rays from elements below atomic number 20 (e.g. calcium)
Kb Lb purges are often used. The two most common purge methods are the vacuum
K shell system and helium purge. Vacuum systems are preferred for analysis of solids
L shell or pressed pellets, while helium is preferred for analysis of liquids or pow-
M shell dered materials.
N shell
X-ray source
Fig. 31.2 Identification of K and L spectral
lines. To generate X-rays you need:
●● A source of electrons.
Desirable properties of an X-ray source – ●● A means of accelerating the electrons.

●● long-term power stability ●● A target, to stop the electrons.
●● low power output requirements
●● purity of spectral output X-ray sources are available in a variety of forms, for example side win-
●● long life dow X-ray tubes (Fig. 31.5), end window X-ray tubes, and radioisotopes. In the
●● small size. case of X-ray tubes – the most common source of X-rays for laboratory-based
instruments – the applied voltage determines which elements can be excited.
Also, the more power is applied, then the lower the detection limits achievable.
Sample
Detectors
Several types of detectors are available for XRF, including:
Detector
●● Si(Li) detectors.
X-ray ●● PIN (positive–intrinsic–negative) diodes.
source
●● Silicon drift detectors.
Fig. 31.3 Schematic diagram of an EDXRF
system. ●● Proportional counters.
●● Scintillation detectors.
Sample For EDXRF the most common detector has been the Si(Li) detector (Fig. 31.6).
Detector
The principle of operation of a detector for XRF is as follows:
1. A detector is composed of a non-conducting or semiconducting material

between two charged electrodes.
Diffraction
X-ray
2. X-ray radiation ionises the detector material causing it to become conduc-
source tive, momentarily.
Fig. 31.4 Schematic diagram of an WDXRF 3. The newly freed electrons are accelerated towards the detector anode to
system. produce an output pulse.
4. The ionised semiconductor produces electron–hole pairs, the number of
pairs produced being proportional to the X-ray photon energy.
Evaluating spectra
EDXRF can be used to analyse, for example, the chemical composition of
glass either quantitatively or qualitatively. It is important to be able to dif-
ferentiate between peaks due to the elements of interest and other spectral

anomalies including interferences. The types of anomalies found in XRF are

High voltage
cathode complex and varied, but need to be recognised in order to distinguish them
from elemental peaks.
Tungsten
target
Elemental K and L spectral peaks
These are the most common elemental peaks observed in EDXRF spectra. For
example, the L shell electron transition to fill a vacancy in the K shell results in
Beryllium the production of Ka radiation. This is the most frequent transition and hence
window produces the most intense peak. Whereas, when an M shell electron transition
occurs to fill a vacancy in the K shell it results in the production of Kb radia-
X-rays tion, and so on (Fig. 31.2). The ratio of signal intensities between Ka and Kb
is normally 20 : 1.
Fig. 31.5 X-ray tube (side window).
Similarly, if an M shell electron transition occurs to fill a vacancy in the
L shell, it results in the production of L a radiation. Whereas, an N shell elec-
Si(Li) detector
tron transition to fill a vacancy in the L shell produces L b radiation, and so
on (Fig. 31.2). In addition to the elemental peaks, other peaks can also appear
Liquid
nitrogen in spectra.
cooling
Pre-amplifier Scatter
Beryllium window When some of the source X-rays strike the sample they are scattered back at
the detector (often referred to as ‘backscatter’). Two types of scatter can be
Fig. 31.6 Schematic diagram of a Si(Li) identified:
detector.
●● Rayleigh scatter – X-rays from the X-ray tube or target strike the atom with-
out promoting fluorescence. In this scenario, energy is not lost in the colli-
sion process. As a result, this type of scatter appears as a source peak in the
line spectrum. It is sometimes referred to as ‘elastic’ scatter.
●● Compton scatter – X-rays from the X-ray tube or target strike the atom with-
out promoting fluorescence. In this scenario, energy is lost in the collision
Bremsstrahlung – The process of elec- process, and scatter appears as a source peak in the line spectrum, but slightly
trons colliding with atoms in the object lower in energy than the Rayleigh scatter. Compton scatter is often referred
under investigation results in decelera- to as ‘inelastic’ scatter.
tion of electrons and production of an
X-ray photon. The resultant continuum Escape peaks
of energy, often called Bremsstrahlung
In normal operation, X-rays strike the sample and promote elemental fluo-
radiation, appears as a broad band
rescence. However, escape peaks result from the fact that some silicon (Si)
across many energies.
fluorescence at the surface of the detector escapes and is not collected by the
detector. The result is a peak that appears in the spectra at an energy that cor-
responds to silicon, i.e. 1.74 keV.
Interferences – Interferences can be
divided into three categories: Sum peaks
Sum peaks result when two photons strike the detector at exactly the same
1. Spectral interferences.
time. In this situation, fluorescence is captured by the detector, and recog-
2. Environmental interferences. nised as a single photon but at twice its normal energy. Sum peaks can be
3. Matrix interferences. observed in spectra at twice the energy for the element under investigation,
i.e. 2 * element keV.
Spectral interferences
Spectral interferences are peaks in the spectrum, from other sources, that over-
lap with the spectral peak of the element to be analysed. Examples include K
and L line overlaps for sulphur and molybdenum, chlorine and rhodium, and

arsenic and lead. In addition, adjacent element overlap can occur, examples
being aluminium and silicon, sulphur and chlorine, and potassium and calcium.
In these situations, it is the resolution of the detector that determines the sig-
nificance of the overlap.
Environmental interferences
Lighter elements, such as those between sodium and chlorine in the Periodic
Table, emit weak X-rays, whose signal can be reduced by air. The remedy is
to either (a) purge the instrument with an inert gas such as helium (as helium
is less dense than air, it results in less attenuation of the signal) or (b) evacuate
air from the sample chamber via a vacuum pump. The removal of air from the
sample chamber also has additional benefits, such as the elimination of spectral
interferences resulting from argon which is present in small quantities in air,
has a spectral overlap with chlorine.
Fe
Fe Ca
Ca
Matrix interferences
Fe
Ca
Fe
Ca
Two types of matrix interferences can result in absorption or enhancement
effects. In absorption, any element can absorb or scatter the fluorescence of
the element of interest, whereas in enhancement characteristic X-rays of one
element can excite another element in the same sample resulting in signal
enhancement. These matrix interferences can be mathematically corrected by
Detector
X-ray the use of influence coefficients, or alpha corrections. This is demonstrated in
source Fig. 31.7 where the incoming source X-ray causes iron in the sample to fluo-
resce. The resultant iron fluorescence is sufficient in energy to cause calcium to
Fig. 31.7 Absorption-enhancement effects fluoresce. In this situation calcium is detected, but iron is not. The resultant sig-
in XRF. nal responses are proportional to the concentrations of both calcium and iron.
Sample preparation
XRF is often regarded as requiring minimal or no sample preparation. How-
ever, for reproducible results, consideration needs to be given to the type and
form of sample to be analysed. Often the sample to be analysed can be placed
directly in a sample cup holder in the XRF instrument. A typical sample cup is
normally 2–4 cm in diameter, and is no higher than it is wide. In addition, the
sample cup holder can be obtained in a variety of configurations, including as
a single open-ended cup (the obvious benefit being that it requires no cover on
the cup) or a double open-ended cup (allows flexibility in terms of selecting
the cover for the sample, e.g. Mylar™ film). You should always use covers for
solution or powder samples, to prevent contamination and/or loss of material.
The most common window (i.e. sample support) film is Mylar™. This
has the combined benefits of low cost and high tensile strength, allowing it to
be produced in a thin form (down to 1.5 mm), and thereby leading to highly
reproducible results. However, Mylar™ has poor resistance to acids, preventing
its use in many applications where acid may have been used in the prepara-
tion of the sample. Other materials used include polypropylene, polycarbonate,
polyimide (Kapton™) and Teflon™. It is also possible to use cups with solid
beryllium windows for analysis of very light elements.
Sample preparation for liquid samples

Liquid samples can be analysed directly by three-quarters filling the sample cup
and placing it in the EDXRF spectrometer. However, problems can arise due to
the liquid sample evaporating, stratifying (forming layers) and/or precipitating

Box 31.1 How to avoid problems with liquid samples in XRF
1. Reduce evaporation of liquid samples by preparing it may be necessary to separate the two liquids and
and analysing samples immediately. Evaporation can analyse them separately. Alternatively, mix the solu-
be reduced by placing a cover over the cup holder, tion rapidly to form an emulsion and then analyse
though this can lead to bulging of the window film, immediately.
resulting in poor reproducibility of measurement.
Some manufacturers produce baffled cups to help to 3. If precipitation occurs, remove the liquid (superna-
reduce evaporation. tant) from the precipitate using a pipette. This then
allows analysis of both liquid and solid fractions.
2. Mixtures or immiscible liquids can give rise to strati- Alternatively, rapidly mix the liquid sample and the
fication problems. In order to assist in their analysis precipitate prior to analysis.
(see Box 31.1). In addition, the liquid sample itself may react with the window
cup film or be absorbed by it. In order to minimise these effects, it is appropri-
ate to prepare all liquid samples immediately before analysis.
Alternative procedures for liquid samples include the addition of a solidify-
ing agent to the liquid sample (e.g. cellulose, alumina or gelatin) to form a solid
support. Alternatively, the sample can be presented as a thin film by applying
the liquid sample to the surface of a support material (e.g. filter paper or the
sample cup itself). The liquid sample can then be analysed directly, wet, in the
case of the filter paper, or dry by evaporating the sample directly in the cup. A
variation on this approach is to use an ion-exchange resin to pre-concentrate the
elements from the liquid sample, filter the resin (now containing the elements)
and analyse this directly.
(a) Structure of cup (b) Cup with sample Sample preparation for solid samples
Solid objects can be placed directly into the sample chamber of the XRF spec-
Fig. 31.8 Sample cup arrangement.
trometer. However, several factors can influence the analysis of solid samples
including surface roughness, particle shape and size, homogeneity, particle
distribution, and mineralistion.
Loose powders can simply be analysed by filling a sample cup to
approximately three-quarters without any need for additional sample prepa-
ration. However, while this approach will give results, it has significant
shortcomings due in part to the heterogeneity of most samples and to varia-
tion in particle size of most loose powders. Figure 31.8 shows a sample cup
(a) Structure of cup (b) Cup with sample where the loose powder sample is supported between a sample film attached
Fig. 31.9 Alternative cup arrangement.
to the sample cup and a semi-permeable (microporous) filter. An alternate
approach is shown in Fig. 31.9. In each case the sample cups are reusable
when thoroughly cleaned.
Pressed pellets are prepared by pressing loose powders filled in a ring
or cup using a set of dies and a press machine. There are two types of dies,
namely flat disc (Fig. 31.10) and cylinder types (Fig. 31.11). The type to be
used depends on the characteristic of the powder sample. Ease of pellitisation
depends on sample characteristics and grain size, and can be improved by
sufficient pellitisation. Mixing the powder sample with a forming agent (i.e. a
“binder”) is another solution if pelletization is difficult (Fig. 31.12).
Powders which are difficult to be formed into pellets can be pellitised by
mixing binder with the sample ideally by pulverisation. Without binder, fine
Fig. 31.10 Sample preparation with flat powder particles may fall off or scatter from the pellet surface and cause con-
type dies. tamination of the spectrometer’s sample chamber in vacuum mode. Powders

which particles are spherically shaped such as SiO2 or burned ash are difficult
to pellitise. Mixing ratio of sample to binder is typically 10(sample) : 1(binder)
or 10 : 2. It is necessary to determine the purity of the binder as it is necessary
to select a binder which does not include the elements to be analysed. Binders
typically used are wax types called Spectro Blend®, polystyrene based powders,
or boric acid and cellulose powders. Addition of binder allows pellitisation
of powders difficult to form, but accurate weighing and complete mixing is
essential to minimise analysis errors. An example of pressed pellets is shown
in Fig. 31.13.
Manual press machines are available, capable of delivering a 300 kN max-
Fig. 31.11 Sample preparation with cylin- imum load (Fig. 31.14). The press can be used for pellitisation with flat and
der type dies. cylinder type dies. The procedure for preparation of a pressed pellet sample is
shown in Box 31.2.
When pellitising samples, there is a possibility that contamination can
occur due to previously pellitised sample remains on the die surface. There-
fore, it is recommended to clean the die surface every time before pellitisation
and to prepare samples starting with lower concentrations. Powder sticking to
the die can be prevented by placing a film in between. Use of the film is effec-
tive not only to minimize contamination but also for samples such as iron or
titanium oxides which cannot be easily pellitised since a significant amount
of powder can stick to the die surface. Sample films such as polypropylene or
Fig. 31.12 Process for making pressed pellet. polyester can be used as film for pellitisation. If a sample needs to be repetel-
lised due to breakage, contamination due to ring or cup material may occur.
Environmental application
An environmental soil sample was subjected to EDXRF. The soil sample was
prepared by air-drying it for 48 h prior to grinding and sieving. The sample
was prepared as indicated in Box 31.2. The pelletised soil sample was then
analysed. The results, shown in Fig. 31.15 as a plot of energy against signal,
identify the range of elements present in the sample. By calibration of the
EDXRF the elements identified can be converted into quantitative data.
Pressed samples
Fig. 31.13 Pressed samples.
Mn
Signal
Ni
Cr Cu
Ca
K Fe Zn Pb
Energy (keV)
Fig. 31.14 A manual press. Fig. 31.15 Representative EDXRF trace of a soil sample.

Box 31.2 How to prepare a loose powder sample for XRF analysis
1. Dry and grind the loose powder sample to a particle 4 g. It is important to keep the weight constant to
size corresponding to 300–400 mesh (15935 mm) or allow consistent results.
better.
5. Place a polished pellet over the sample to produce a
2. Mix a portion of the dried powder with a binder, for smooth finish.
example, paraffin or cellulose (10 : 2 w/w). The binder
helps to hold the finished pellet together. 6. Insert the plunger and position the die in the press.
Press the pellet at a pressure of 5–30 tonnes main-
3. Clean the die. This can be done by wiping with meth- tained for approximately 60 s.
anol or another solvent.
7. Remove the pellet from the die, taking care not to
4. Insert the backing (i.e. an aluminium cap) into the die crack it in the process. The pellet is then ready for
and accurately weigh the sample added, for example, analysis.

Beckhoff, B., Kanngiesser, B., Langhoff, N., Wedell, R. and Kellner, R., Mermet, J.M., Otto, M., Valcarcel, M. and
Wolff, H. (2006) Handbook of Practical X-Ray Fluores- Widmer, H.M. (2004) Analytical Chemistry: A Modern
cence Analysis. Springer, Berlin. Approach to Analytical Science, 2nd edn. John Wiley &
Buhrke, V.E., Jenkins, R. and Smith, D.K. (1998) A Prac- Sons Ltd, Chichester.
tical Guide for the Preparation of Specimens of X-Ray Rouessac, F. and Rouessac, A. (2007) Chemical Analy-
Fluorescence and X-Ray Diffraction Analysis. John sis: Modern Instrumentation Methods and Techniques,
Wiley & Sons Ltd, Chichester. 2nd edn. John Wiley & Sons Ltd, Chichester.
Christian, G.D., Dasgupta, P.K. and Shrug, K.A. (2014) Schwedt, G. (1997) The Essential Guide to Analytical
Analytical Chemistry, 6th edn. John Wiley & Sons Ltd, Chemistry. 2nd edn. John Wiley & Sons Ltd, Chichester.
Chichester. Vandecasteele, C. and Block, C.B. (1993) Modern Methods
Jenkins, R. (1999) X-ray Fluorescence Spectrometry, for Trace Element Determination. John Wiley & Sons
2nd edn. John Wiley & Sons Ltd, Chichester. Ltd, Chichester.
Study exercises
31.1 Explain the principle of X-ray fluorescence spec- stepwise protocol explaining how to prepare the
troscopy. Ask a colleague to listen to your expla- soil sample and analyse it by XRF. Ask another
nation and provide feedback. student to evaluate your protocol and provide
31.2 Test your knowledge. What are the main types you with written feedback – either simply by
of interferences associated with XRF and explain reading through your protocol, or by trying it out
how they occur and how to avoid them. as part of a class exercise. (Check with a member
of staff before you attempt this in a laboratory.)
31.3 Write a protocol for analysing a sample of soil.
After reading this chapter, prepare a detailed

32 Chromatography
Chromatography is used to separate the individual components of a mixture

Chromatography is often a three-way
on the basis of differences in their physical characteristics, for example molec-
compromise between:
ular size, shape, charge, volatility, solubility and/or adsorption properties. The
1. separation of analytes; essential components of a chromatographic system are:
2. time of analysis;
3. volume of eluent. ●● A stationary phase, where a solid, a gel or an immobilised liquid is held
by a support matrix.
●● A chromatographic bed: the stationary phase may be packed into a glass or
metal column, spread as a thin layer on a sheet of glass or plastic, or adsorbed
on cellulose fibres (paper).
Selecting a separation method – it is ●● A mobile phase, either a liquid or a gas which acts as a solvent, carrying the
often best to select a technique that sample through the stationary phase and eluting from the chromatographic bed.
involves direct interaction between the
●● A delivery system to pass the mobile phase through the chromatographic bed.
substance(s) and the stationary phase
(e.g. ion-exchange chromatography), ●● A detection system to visualise the test substances.
owing to their increased capacity and
The individual substances in the mixture interact with the stationary phase to
resolution compared with other meth-
different extents, as they are carried through the system, enabling separation
ods (e.g. partition or gel permeation
to be achieved.
chromatography) where the analytes
are not bound to the stationary phase.
Key Point In a chromatographic system, those substances

which interact strongly with the stationary phase will be retarded
solid stationary
phase with
to the greatest extent, while those which show little interaction
polar groups
d– d+ will pass through with minimal delay, leading to differences in
d+
d– distances travelled or elution times.
d–
d+ d– polar molecules
d+ are held by
d+ d– dipole–dipole
d–
d– interaction
d+ d+ Separation methods
non-polar molecules
pass rapidly Chromatography is subdivided according to the mechanism of interaction of
through
the column the solute with the stationary phase.
flow of mobile phase
Fig. 32.1 Adsorption chromatography Adsorption chromatography

(polar stationary phase). This is a form of solid–liquid chromatography. The stationary phase is
a porous, finely divided solid which adsorbs molecules of the mixture on
its surface by dipole–dipole interactions, hydrogen bonding and/or van der
liquid Waals’ interactions (Fig. 32.1). The range of adsorbents is limited to polysty-
stationary phase non-polar molecules
preferentially dissolve rene-based resins for non-polar molecules and silica, aluminium oxide and
in the stationary calcium phosphate for polar molecules. Most adsorbents must be activated by
phase
heating to 110–120 °C before use, since their adsorptive capacity is signifi-
cantly decreased if water is adsorbed on the surface. Adsorption chromatog-
polar molecules
raphy can be carried out in column (p. 263) or thin-layer (p. 260) form, using
inert pass rapidly a wide range of organic solvents.
support through
matrix the column
Partition chromatography
flow of This is based on the partitioning of a substance between two liquid phases,
mobile phase
in this instance the stationary and mobile phases. Substances which are more
Fig. 32.2 Liquid-liquid partition chromatog- soluble in the mobile phase will pass rapidly through the system while those
raphy, e.g. reversed-phase HPLC. which favour the stationary phase will be retarded (Fig. 32.2). In normal phase

Chromatography
partition chromatography the stationary phase is a polar solvent, usually water,

Maximising resolution in IEC – keep
supported by a solid matrix (e.g. cellulose fibres in paper chromatography) and
your columns as short as possible.
the mobile phase is an immiscible, non-polar organic solvent. For reversed-
Once the sample components have
phase partition chromatography, the stationary phase is a non-polar solvent
been separated, they should be eluted
(e.g. a C18 hydrocarbon, such as octadecylsilane) which is chemically bonded
as quickly as possible from the column
to a porous support matrix (e.g. silica), while the mobile phase can be chosen
in order to avoid band broadening
from a wide range of polar solvents, usually water or an aqueous buffered
resulting from diffusion of sample ions
solution containing one or more organic solvents (e.g. acetonitrile). Solutes
in the mobile phase.
interact with the stationary phase through non-polar interactions and so the
least polar solutes elute last from the column. Solute retention and separa-
sample cations displace mobile counterions tion are controlled by changing the composition of the mobile phase (e.g. %
(e.g. H+) and are retained on the column
v/v acetonitrile). Reversed-phase high-performance liquid chromatography
stationary phase is a
cation-exchange – (RPHPLC, p. 265) is used to separate a broad range of non-polar, polar and
– +
resin +
–
ionic molecules, including environmental compounds (e.g. phenols) and phar-
– –+ – + maceutical compounds (e.g. steroids).
anionic / neutral
+– – – solutes are
not retained Ion-exchange chromatography (IEC)
–
– – Here, separations are carried out using a column packed with a porous matrix
–
which has a large number of ionised groups on its surfaces, i.e. the sta-
flow of mobile phase tionary phase is an ion-exchange resin. The groups may be cation or anion
exchangers, depending upon their affinity for positive or negative ions. The
Fig. 32.3 Ion-exchange chromatography net charge on a particular resin depends on the pKa of the ionisable groups
(cation exchanger).
and the pH of the solution, in accordance with the Henderson–Hasselbalch
small molecules have
equation (p. 117).
access to the whole matrix, For most practical applications, you should select the ion-exchange resin
taking the longest to pass and buffer pH so that the test substances are strongly bound by electrostatic
through the column
medium-sized molecules attraction to the ion-exchange resin on passage through the system, while the
stationary may have access other components of the sample are rapidly eluted (Fig. 32.3). You can then
phase is a to the larger pores
microporous elute the bound components by raising the salt concentration of the mobile
gel phase, either stepwise or as a continuous gradient, so that exchange of ions
large molecules are of the same charge occurs at oppositely charged sites on the stationary phase.
totally excluded Weakly bound sample molecules will elute first, while more strongly bound
from the pores,
passing rapidly molecules will elute at a higher concentration.
through Computer-controlled gradient formers are available: if two or more com-
the column
ponents cannot be resolved using a linear salt gradient, an adapted gradient
flow of
can be used in which the rate of change in salt concentration is decreased
mobile phase over the range where these components are expected to elute. IEC can be
used to separate mixtures of a wide range of anionic and cationic compounds.
Fig. 32.4 Gel permeation chromatography. Electrophoresis (Chapter 33) is an alternative means of separating charged
Table 32.1 Fractionation ranges of selected molecules.
GPC media
Gel permeation chromatography (GPC) or gel filtration
Mr Medium
Here, the stationary phase is in the form of beads of a cross-linked gel con-
50–1 000 Sephadex G15, Biogel P-2 taining pores of a discrete size (Fig. 32.4). The size of the pores is controlled
1 000–5 000 Sephadex G-25 so that at the molecular level, the pores act as ‘gates’ that will exclude large
molecules and admit smaller ones (Table 32.1). However, this gating effect is
1 500–30 000 Sephadex G-50, Biogel P-10
not an all-or-nothing phenomenon: molecules of intermediate size partly enter
4 000–150 000 Sephadex G-100 the pores. A column packed with such beads will have within it two effective
5 000–250 000 Sephadex G-200 volumes that are potentially available to sample molecules in the mobile phase,
20 000–1 500 000 Sephacryl S 300 i.e. Vi, the volume surrounding the beads and Vii, the volume within the pores.
If a sample is placed at the top of such a column, the mobile phase will carry
60 000–20 000 000 Sepharose 4B
the sample components down the column, but at different rates according to

Chromatography
their molecular size. A very large molecule will have access to all of Vi but
Using a gel permeation system – keep
to none of Vii, and will therefore elute in the minimum possible volume (the
your sample volume as small as possi-
‘void volume’, or V0, equivalent to Vi). A very small molecule will have access
ble, in order to minimise band broaden-
to all of Vi and all of Vii, and therefore it has to pass through the total liquid
ing due to dilution of the sample during
volume of the column (Vt, equivalent to Vi + Vii) before it emerges. Molecules
passage through the column.
of intermediate size have access to all of Vi but only part of Vii, and will elute
at a volume between V0 and Vt, in order of decreasing size depending on their
access to Vii.
Elution of substances from an affinity
Cross-linked dextrans (e.g. Sephadex®), agarose (e.g. Sepharose®) and
system – make sure that your elution
polyacrylamide (e.g. Bio-gel®) can be used to separate mixtures of macro-
conditions do not affect the interaction
molecules, particularly enzymes, antibodies and other globular proteins. Selec-
between the ligand and the stationary
tivity in GPC is solely dependent on the stationary phase, with the mobile phase
phase, or you may elute the ligand from
being used solely to transport the sample components through the column.
the column.
Thus, it is possible to estimate the molecular mass of a sample component
by calibrating a given column using molecules of known molecular mass and
similar shape. A plot of elution volume (Ve) against log10 molecular mass is
approximately linear. A further application of GPC is the general separation
Remember that you cannot quantify a of components of low molecular mass and high molecular mass, for example
particular analyte without first identify- ‘desalting’ a protein extract by passage through a Sephadex® G-25 column is
ing it: the presence of a single peak on faster and more efficient than dialysis.
a chromatogram does not prove that a
single type of analyte is present.
The chromatogram
A plot of the detector response present at the column outlet as a function of
time is called a chromatogram (Fig. 32.5). The time from injection of the sam-
ple until the peak elutes from the column is called the retention time, tr. The
amount of compound present for a given peak can be quantified by measuring
the peak height or area (most useful) and comparing it with the response for a
known amount of the same compound.
The aim of any chromatographic system is to resolve a number of compo-
nents in a sample mixture, i.e. to ensure that individual peaks do not overlap
or coincide. To achieve this you need to consider several important factors:
capacity factor, separation factor or selectivity, column efficiency and asym-
metry factor.
Capacity factor, k′ This is a more useful measure of peak retention that

retention time, as it is independent of column length and flow rate. To calcu-
late k′, you need to measure column dead time, to. This is the time it takes an
unretained component to pass through the column without any interaction with
the stationary phase. It is the time taken from the point of sample injection until
the first disturbance in the base line caused by the unretained component. The
capacity factor for other components can then be calculated according to the
following equation:
tR - to
k′ = [32.1]
to
Learning from experience – if you are
unable to separate your molecule using Separation factor, a The separation factor, or selectivity, identifies when the
a particular method, do not regard this peaks elute relative to each other. It is defined for two peaks as the ratio of the
as a failure, but instead think about capacity factors (k 2= 7 k 1= ):
what this tells you about either the sub- k 2= tR, 2 - to
stance(s) or the sample. a = = = [32.2]
k1 tR, 1 - to

Chromatography
where tR,1 and tR,2 are the retention times of peak 1 and peak 2, respectively. If
Column efficiency – several equations
two peaks are present the separation factor must be greater than one to achieve
can be used to calculate the column
an effective separation.
efficiency. Select the correct equation
based on the information available.
Column efficiency (plate number), N An additional parameter used to
characterise a separation system is the plate number, N. It represents, in gen-
eral terms, the narrowness of the peak and is often calculated using one of the
following equations:
tR 2
N = 5.54¢ ≤ [32.3]
w0.5
A tR 2
w0.5 h N = 16¢ ≤ [32.4]
wb
start
tR.h 2
tR N = 2p¢ ≤ [32.5]
wb A
0.5h
to
where tR is the retention time of the peak, w0.5 and wb are the peak widths at
time
half height and base, respectively, and h and A are the peak height and peak
area, respectively (Fig. 32.5).
Fig. 32.5 Peak characteristics in a chroma- For a compound emerging from a column of length L, the number of the-
tographic separation, i.e. a chromatogram. oretical plates, N, can be expressed as:
For symbols, see eqns [32.1, 32.3–32.5].
L
N = [32.6]
H
where H is the plate height (or height equivalent to a theoretical plate). In

10% of peak height general, chromatographic columns with larger values of N give the narrowest
height peaks and generally better separation.
a b Asymmetry factor, As The plate number, N, assumes that the peak shape is
Gaussian, but in practice this is rare. It is more likely that the peak is asymmet-
time rical, i.e. it ‘tails’. This is quantified using the asymmetry factor, As, calculated
as shown in Fig. 32.6.
Fig. 32.6 Peak asymmetry.
A vertical line is drawn between the peak maximum and the base line.
At 10% of the peak height, the width of the peak to the leading edge and the
trailing edge is measured (a and b in Fig. 32.6). The asymmetry factor is then
calculated as follows:
b
As = [32.7]
a
In general, As values between 0.9 and 1.2 are acceptable. If As 7 1 peak

tailing is in evidence; if As 6 1 peak fronting is evident. The practical impact
of peak tailing or fronting is that adjacent peaks are not as well separated
as they would be if they were symmetrical, leading to difficulties in peak
quantitation.
4 8 12 16 20 24 28 32
time/mins Resolution
Fig. 32.7 A multicomponent chromatogram. It is often important to be able to separate a large number of compounds. A
Separation of many compounds, some well visual inspection of the chromatogram (Fig. 32.7) will usually indicate whether
resolved, e.g. peaks at 12–13 min, and oth- the separation is appropriate. It is desirable that the valley between adjoining
ers that are not, e.g. peaks at 24–25 min. peaks returns to the base line and resolution is a quantitative measure of the

Chromatography
separation. The influence of k′, a and N on resolution, R, is shown in the fol-

lowing expression:
1. initial
2N k′ a - 1
R = * * [32.8]
4 k′ + 1 a
2. vary k' Three conditions must be satisfied in order to achieve some degree of resolution:
1. Peaks have to be retained on the column (k′ 7 0).
3. increase N
2. Peaks have to be separated from each other (a 7 1).
3. The column must develop some minimum value of N.
These different effects and their influence on resolution are shown in
4. increase c Fig. 32.8, using high-performance liquid chromatography (HPLC) as an
inject V0 V example (see p. 265).
Fig. 32.8 Influence of k′, a and N on 1. Initial conditions result in inadequate separation of the two components.
resolution.
2. Effect of varying the capacity factor, k′: from an initial mobile phase of
50% methanol : 50% water (v/v) two scenarios are possibile. Firstly, on
the left-hand side, the influence of increasing the percentage of organic
solvent (70% methanol: 30% water) allows a faster through-put, but the
peaks are unresolved. Secondly, on the right-hand side, the percentage
organic solvent is reduced (30% methanol : 70% water) allowing the com-
ponents to remain in the system for a longer time, giving some separation,
but causing peak broadening. This is the easiest change to make and will
affect resolution. As a guide, a two- to three-fold change in k′, will result
for each 10% change in mobile phase composition.
3. Effect of increasing the plate number, N: reducing the particle size of the
HPLC packing from 5 mm to 3 mm allows a more efficient separation.
It should be noted that the retention times of the peaks are not altered
(from the initial chromatogram) provided the stationary phase is not
altered. Alternatively N can be increased by placing columns in series
with one another. However, you should note from eqn [32.8] that R has
a square-root dependence on N, i.e. a four-fold increase in N is required
to double R.
4. Increase separation factor, a: resolved peaks can be obtained by changing
the mobile phase (e.g. methanol to acetonitrile) or the column stationary
phase (e.g. C18 to C8). Unfortunately this is the least predictable approach.
Detectors – a range of detectors are

Detectors
available for chromatography. Mass After separating the components of the mixture it is necessary to detect them.
spectrometry is the only detector com- A description of a range of detectors suitable for chromatography is described
patible with GC or HPLC. (p. 258). As chromatography is often used as a quantitative technique it is
essential to be familiar with the following terms:
●● Universal detector: this responds to all compounds eluting from the column,
irrespective of their composition.
●● Selective/specific detector: this responds to certain elements or functional
groups. This is a useful approach if the components of the mixture are known.
●● Sensitivity: the ratio of detector signal to sample size (or detector response
per amount of sample).

Chromatography
●● Minimum detectable level (MDL): the amount of sample in which the peak
height is at least twice the noise height.
●● Linear dynamic range: the concentration range of the sample that is detecta-
ble and where the detector response is linear (between the MDL and detector
saturation).
Types of chromatographic system

Chromatographic systems can be categorised according to the form of the chro-
matographic bed, the nature of the mobile and stationary phases and the method
of separation.
sample
pressure regulator
injection
Gas chromatography
port
In gas chromatography (GC), a gaseous solute (or the vapour from a volatile
detector
liquid) is carried by the gaseous mobile phase. In gas–liquid partition chroma-
microcomputer /
readout tography, the stationary phase is a non-volatile liquid coated on the inside of
carrier
gas the column or on a fine support. In gas–solid adsorption chromatography, solid
supply particles that adsorb the solute act as the stationary phase.
The typical components of a gas chromatograph are shown in Fig. 32.9. A
volatile liquid is injected through a septum into a heated port, which volatilises
column
oven
the sample. A gaseous mobile phase carries the sample through the heated
column, and the separated components are detected and recorded. Two types
Fig. 32.9 Components of a GC system. of column are available: packed and capillary. Open tubular capillary columns
offer higher resolution, shorter analysis time and greater sensitivity than packed
columns, but have lower capacity for the sample.
Sample injection
Types of capillary column
Samples are injected onto the ‘top’ of the column, through a sample injection
●● Wall-coated open tubular (WCOT)
port containing a gas-tight septum. The two common sample injection methods
column: liquid stationary phase on
inside wall of column.
for capillary GC are:
●● Support-coated open tubular (SCOT) 1. Split/splitless injector: in the split mode only a portion of the injected
column: liquid stationary phase sample (typically, 1 part in 50) reaches the column. The rest is vented to
coated on solid support attached to waste. A split injector is used for concentrated samples ( 7 0.1 mg mL-1
inside wall of column.
for FID; see p. 258). In the splitless mode all the sample volume injected
●● Porous layer open tubular (PLOT)
column: solid stationary phase on
passes through to the column. It is used, in this mode, for trace samples
inside wall of column. ( 6 0.1 mg mL-1 for FID).
2. Cold on-column injector: all the sample is injected onto the column. It
is used for thermally unstable compounds and high-boiling solvents.
In both cases, a syringe (1 mL) is used to inject the sample. Examples of each
type of injection system are shown in Fig. 32.10. The procedure for injection of
a sample is shown in Fig. 32.11. In Fig. 32.11(a) the syringe is filled with the
sample/standard solution (typically 0.5 mL). Then the outside of the syringe
Applications of gas–liquid chromatog- is wiped clean with a tissue (Fig. 32.11(b)). The syringe is placed into the
raphy – GLC is used to separate vola- injector of the gas chromatograph (Fig. 32.11(c)) and, finally, the plunger on
tile, non-polar compounds: substances the syringe is depressed to inject the sample (Fig. 32.11(d)). The procedure
with polar groups must be converted to for the preparation of a series of calibration solutions is shown in Box 32.1.
less polar derivatives prior to analysis, The column
in order to prevent adsorption on the
column, resulting in pool resolution and
Modern GC uses capillary columns (internal diameter 0.1–0.5 mm) up to 60 m
peak tailing.
in length. The stationary phase is generally a cross-linked silicone polymer,
coated as a thin film on the inner wall of the fused silica (SiO2) capillary

Chromatography
syringe
PTFE block
syringe
to purge
valve ball valve
septum
syringe needle
glass liner
column end
carrier
gas inlet
column
compressed
air
for cooling
inside the oven
carrier to split valve
gas inlet
syringe needle
(a) (b)
Fig. 32.10 Sample introduction in GC: (a) split/splitless injector; (b) on-column injector.
Fig. 32.12: at normal operating temperatures. This behaves in a similar man-

Analysing compounds by GC in the
ner to a liquid film, but is far more robust. Common stationary phases for GC
split mode – make sure no hazardous
are shown in Fig. 32.13. The mobile phase (‘carrier gas’) is usually nitrogen
materials enter the laboratory atmos-
or helium. Selective separation is achieved as a result of the differential parti-
phere through the split vent. A charcoal
tioning of individual compounds between the carrier gas and silicone polymer
split-vent trap may be required to elim-
phases. The separation of most organic molecules is influenced by the temper-
inate potential hazards.
ature of the column, which may be constant during the analysis (‘isothermal’ –
usually 50–250 °C) or, more commonly, may increase in a pre-programmed
manner (e.g. from 50 °C to 250 °C at 10 °C per min).
Selecting an appropriate column for capillary GC is a difficult task and
one which is usually left to the technician. However, it is important to be
aware of some general issues and what influence they can have on the sep-
aration. The column internal diameter can affect both resolution and speed
of analysis. Smaller internal diameters columns (0.25 mm i.d.) can provide
good resolution of early eluting peaks (Fig. 32.14(a)). However, the problem
is that the analysis times of the eluting components may be longer and that the
linear dynamic range may be restricted. In contrast, larger internal diameter
columns (0.53 mm i.d.) provide less resolution for early eluting compounds
(Fig. 32.14b), but this is reflected in shorter analysis times and a greater lin-
ear dynamic range. This type of column may provide sufficient resolution for
the analysis of complex mixtures. Fig. 32.14 illustrates the effects of column
internal diameter.

Chromatography
100% dimethylsiloxane:
the least polar bonded phase. Used for boiling point
separations (solvents, petroleum products, etc.).
CH3
Si O
CH3
(a) (b)
95% dimethylsiloxane–5% diphenylypolysiloxane:

a non-polar phase. Used for separation of environmental
samples, e.g. polycyclic aromatic hydrocarbons.
CH3
Si O Si O
CH3
(c) (d) 5% 95%
Fig. 32.11 Sample injection in GC. (a) Fill the syringe, (b) wipe clean the Fig. 32.13 Common stationary phases for
outside of the syringe needle, (c) place the syringe needle into the injector capillary GC.
and (d) depress the plunger on the syringe to inject the sample.
Polyimide
coating
Fused-silica
Stationary
phase
Fig. 32.12 Schematic diagram of a GC column.
Box 32.1 H
ow to prepare a set of five calibration solutions in the concentration range
0910 mg mL −1 (mg L -1)
Assuming that we are starting with a 1000 mg mL-1 so 20 mg 2-chlorophenol was placed in the 10.00 mL
stock solution of a particular organic compound (e.g. volumetric flask. So,
2-chlorophenol) you will need the following: 6 * 10.00 mL 20 mg
grade A volumetric flasks and a syringe (09100.00 mL). = 2 mg mL-12 @chlorophenol[32.10]
10 mL
1. Ensure that all the glassware is clean (see p. 68).
You now have a 2 mg mL-1 calibration solution of
2. Add ?9 mL of organic solvent (e.g. dichloromethane) 2-chlorophenol.
to a 10.00 mL with dichloromethane.
5. Similarly transfer 0, 40.00, 60.00, 80.00 and 100.00 mL
3. Quantitatively transfer 20.00 mL of the stock solution volumes into separate volumetric flasks and dilute
into the 10.00 mL volumetric flask. Inject the solution to 10.00 mL with dichloromethane and label as 0,
from the syringe below the surface of the dichlorometh- 4, 6, 8 and 10 mg mL-1 2-chlorophenol calibration
ane. Then, dilute to 10.00 mL with dichloromethane. solutions.
4. What is the concentration of this new solution? 6. Take the 0, 2, 4, 6, 8 and 10 mg mL-1 2-chlorophe-
Remember that we started with an initial 1000 mg mL-1 nol calibration solutions to the chromatograph for
2-chlorophenol stock solution. analysis.
1000 mg
* 20 * 10-3 mL = 20 mg 2@chlorophenol
mL
[32.9]

Chromatography
(a) 30 m × 0.25 mm i.d. Another important column effect is the length of the column and the influ-
ence this can have on the resolution of eluting components. It was previously
4 shown (p. 254) that resolution was influenced by k′, a and N. Substituting eqn
6
[32.6] into eqn [32.8] produces the following equation:
9
1 L k′ a - 1
1
10
R = * * [32.11]
4Ah k′ + 1 a
3 8
5 7 The importance of this equation can be shown by considering the influence on
resolution, R, of column length, L. Under isothermal analysis conditions, i.e. the
2 11
same column temperature, the retention of eluting compounds is more depend-
ent upon column length. For example, doubling the column length doubles the
analysis times and increases the resolution by 41%. This is shown in Fig. 32.15
for the analysis of phenols. In contrast, under temperature-programmed anal-
ysis (e.g. 130 °C to 250 °C at 4° C min -1) the retention time of eluting com-
4 8 12 16 20 24 ponents is more dependent on temperature. For example, doubling the column
min
length has minimal effect on analysis times. This is shown in Fig. 32.16 for the
analysis of bacterial acid methyl esters.
(b) 30 m × 0.53 mm i.d.
9
GC detectors
4 6 The output from the GC column is monitored by a detector. The most com-
10 monly used detectors for GC analysis of organic molecules are as follows.
1 The flame ionisation detector (FID) is particularly useful for the anal-
ysis of a broad range of organic molecules. It involves passing the exit gas
8 stream from the column through a hydrogen flame that has a potential of
2 more than 100 V applied across it (Fig. 32.17). Most organic compounds, on
3 5 passage through this flame, produce ions and electrons that create a small
7
11
current across the electrodes, and this is amplified for measurement purposes.
The FID is very sensitive (typically down to ≈0.1 pg), with a linear response
over a wide concentration range. One drawback is that the sample is destroyed
during analysis.
The thermal conductivity detector (TCD) is based on changes in the ther-
mal conductivity of the gas stream brought about by the presence of separated
4 8 12 16 20 24 sample molecules. The detector elements are two electrically heated platinum
min wires, one in a chamber through which only the carrier gas flows (the refer-
ence detector cell), and the other in a chamber that takes the gas flow from the
Fig. 32.14 Influence of GC column inter-
column (the sample detector cell). In the presence of a constant gas flow, the
nal diameter on separation: 1. phenol;
2. 2-chlorophenol; 3. 2-nitrophenol;
temperature of the wires (and therefore their electrical resistance) is dependent
4. 2,4-dimethylphenol; 5. 2,4-dichloro- on the thermal conductivity of the gas. Analytes in the gas stream are detected
phenol; 6. 4-chloro-3-methylphenol; 7. by temperature-dependent changes in resistance based on the thermal conduc-
2,4,6-trichlorophenol; 8. 2,4-dinitrophenol; tivity of each separated molecule; the size of the signal is directly related to
9. 4-nitrophenol; 10. 2-methyl-4,6-dinitro- concentration of the analyte.
phenol; 11. pentachlorophenol. The advantages of the TCD include its applicability to a wide range of
organic and inorganic molecules and its non-destructive nature, since the sam-
ple can be collected for further study. Its major limitation is its low sensitivity
(down to ≈10 ng), compared with other systems.
Solvent selection in GC – choose a The electron capture detector (ECD) is highly sensitive (Fig. 32.18) and is
solvent with a boiling point at least 20 useful for the detection of certain compounds with electronegative functional
°C below that of the first eluting com- groups (e.g. halogens, peroxides and quinones). The gas stream from the col-
pound. Thicker stationary-phase films umn passes over a b@emitter such as 62Ni, which provides electrons that cause
provide longer retention and higher ionisation of the carrier gas (e.g. nitrogen). When carrier gas alone is passing
peak capacity. the b@emitter, its ionisation results in a constant current flowing between two
electrodes placed in the gas flow. However, when electron-capturing sample

Chromatography
30 m × 0.25 mm i.d. 1 60 m × 0.25 mm i.d.

1
2
2
5
4
7
8
6 4
3 13
11 12 13
6 78 11 12
3
10 10
9 9
4 8 12 16 min 4 8 12 16 20 24 28 32 36 min
Fig. 32.15 Influence of column length on analysis time. Analysis of phenols under isothermal conditions: 1. phenol;
2. o-cresol; 3. 2,6-xylenol; 4. p-cresol; 5. m-cresol; 6. o-ethylphenol; 7. 2,4-xylenol; 8. 2,5-xylenol; 9. 2,3-xylenol;
10. p-ethylphenol; 11. m-ethylphenol; 12. 3,5-xylenol; 13. 3,4-xylenol.
30 m × 0.53 mm i.d. 60 m × 0.53 mm i.d.
4 8 12 16 20 24 28 min 4 8 12 16 20 24 28 32 min
Fig. 32.16 Influence of column length on analysis time. Analysis of bacterial acid methyl esters under temperature-
programmed conditions.
molecules are present in the gas flow, a decrease in current is detected. An

Detectors – the most appropriate
example of the application of the ECD is in detecting and quantifying chlorin-
detector depends on the type of chro-
ated pesticides.
matography and the application: ide-
Mass spectrometry (see also p. 355) used in conjunction with GC pro-
ally, the detector should show high
vides a powerful tool for identifying the components of complex mixtures,
sensitivity, a low detection limit and
for example environmental pollutants, synthetic products (Fig. 32.19). The
minimal noise or drift. These terms are
procedure requires computer control of the instrument and for data storage/
defined on p. 254.
analysis. Compounds eluting from the column are bombarded by electrons
(electron impact, EI, mode) causing fragmentation and production of charged
species. These charged species are separated by the mass spectrometer on the
basis of their mass-to-charge ratio. Ions passing through the mass spectrometer

Chromatography
are detected by an electron multiplier tube. The mass spectrometer can be used
in two modes (p. 363): total ion and selected ion monitoring. In the former
mode, the complete mass spectrum of each of the components of the mixture
electrical eluting from the column is recorded. In the latter mode, only ions of specified
output platinum anode
mass-to-charge ratios are detected. Selected ion monitoring offers increased
flame platinum cathode sensitivity and selectivity.
jet assembly
air
input
Liquid chromatography
The basic chromatographic system comprises a stationary phase (adsorbant),
FID body
usually alumina, silica gel or cellulose, through which a mobile phase travels
hydrogen
input (elutes). Separation of a mixture of compounds is achieved by a combination
of the differing ‘adsorption’ and solubility characteristics of the components
on the stationary phase and in the mobile phase respectively.
Liquid chromatography is used both as an analytical method to determine
gas flow
from column
the complexity of mixtures and the purity of compounds, and as a preparative
system for the separation of mixtures. Liquid chromatography is divided into
Fig. 32.17 Components of a flame ion- two general types:
isation detector (FID).
1. Thin-layer chromatography (TLC): in which a glass or plastic plate is
coated with a thin layer of the stationary phase and the mobile phase
ascends the plate by capillary action. TLC is essentially an analytical tool
and preparative TLC has been largely superseded by flash chromatography.
electrical
2. Column chromatography: in which the stationary phase is packed into
output collector electrode a glass column and the mobile phase is passed down the column, either
(anode) by gravity (gravity chromatography) or under low pressure from a pump
or nitrogen cylinder (flash chromatography). These are the preparative
radioactive systems.
foil (63Ni)
Thin-layer chromatography
cathode
The essential components of a TLC system are:
ECD body
●● The stationary phase comprising the layer of adsorbant on a solid backing –
the chromatoplate. Aluminium or plastic-backed chromato-plates are now the
norm having replaced glass plates, which needed to be prepared ‘in-house’.
The chromatoplates (20 cm * 20 cm) can be cut down to the more useful
size (2 cm * 5 cm) for analytical work, using a guillotine. The adsorbant
gas flow often contains a fluorescent compound (ZnS) to enable visualisation of the
from column compounds after elution.
Fig. 32.18 Components of an electron cap-
ture detector (ECD).
syringe chromatogram
ion source detector

Using TLC plates – a plate (2 cm *
5 cm) will hold three sample ‘spots’. oven
computer
Using TLC plates – do not touch the

surface of the TLC plates with your fin- interface
column
gers. Hold them by the edges to prevent vacuum
contamination.
Fig. 32.19 Schematic diagram of a GCMS instrument.

Chromatography
Box 32.2 How to make micropipettes for TLC
1. Heat the middle of an open-ended melting point tube 3. Allow the tube to cool and then break in the centre
at the tip of the hot flame of a microburner until it of the capillary. You now have two micropipettes. If
begins to sag. If the melting point tube is sealed at the capillary is too long, break it near to each end and
one or both ends, carefully break off the sealed end(s) immediately dispose of the fine waste glass into the
wearing gloves for protection. broken-glass bin. Do not leave the waste glass on the
laboratory bench.
2. Quickly remove the tube from the flame and pull
gently, forming a short capillary. Do not pull the tube 4. Make at least 10 micropipettes and store them in a
while it is in the flame. plastic-capped sample tube for future use.
●● The development tank: for plates of (2 cm * 5 cm) a clean, dry beaker (100
watch-glass cover
mL) covered with a watch-glass is ideal. The eluting solvent should be about
3 mm deep and filter paper should be placed in the tank to saturate the tank
TLC plate atmosphere with solvent vapour (Fig. 32.20).
100 mL beaker
●● The application system: a micropipette or a microsyringe to place the solu-
tion of the mixture on the chromatoplate. Micropipettes (Fig. 32.21) are the
two halves of
filter paper more common and Box 32.2 gives the instructions for their preparation.
3 mm of eluent
●● The eluent: finding the eluent, which will give the best separation of the
components of the mixture, is by experiment – you may need to try several
solvents of differing polarity (Table 32.2) or mixtures of solvents to find the
Fig. 32.20 The developing tank for TLC.
best eluent.
●● A visualisation system to be able to see colourless separated components on
Solutions of the mixture – the mixture the chromatogram. If the plate contains a fluorescer, it can be viewed under
must be applied to the plate as a solu- UV light (l = 254 nm) in a special box or cabinet. The ZnS in the stationary
tion. If the solvent for the solution is not phase fluoresces green, whereas the ‘spots’ of separated compounds appear
the same as the eluent to be used, you dark. Alternatively, the plate can be placed in a sealed jar containing a few
must evaporate the solvent from the iodine crystals. The iodine vapour stains the plate light brown and the ‘spots’
plate, before placing it in the eluent. dark brown.
You can express the movement of an individual compound up the TLC plate in
terms of its Rf (relative frontal mobility) value, where:
Finding an eluent – a medium-polarity
solvent such as dichloromethane (DCM) Rf = distance moved by compund/distance moved by solvent[32.12]
is generally a good starting point.
Table 32.2 The elutropic series of solvents for

chromatography
too long Non-polar Light petroleum (b.pt. 40–60 °C)

If end(s) closed –
break off Cyclohexane
break Toluene
break
here here
hold Dichloromethane
capillary Diethylether (ether)
in flame
break AS Ethyl ethanoate (ethyl acetate)
remove INDICATED Propanone (acetone)
from flame to get two
and pull spotters Ethanoic acid (acetic acid)
Polar Methanol
Fig. 32.21 How to make micropipettes.

Chromatography
solvent
front
solvent
authentic front
component spot
compound
Rf of A … 0.452
component spot Rf of B … 0.476
base line
component spot A B A AZB B A A=B B
(a) (b) (c)
Fig. 32.23 Double-spotting technique:

(a) compounds A and B with close Rf val-
ues; (b) figure ‘8’ of double spot shows
base line
mixture that A and B are different; (c) single spot
for double spot shows that A and B could
Fig. 32.22 A thin-layer chromatogram. be the same.
The Rf value is a constant for a particular substance and eluent system

Safety Note Great care must on a specific stationary phase, but variations in chromatographic conditions
be taken when using UV light. Do not adsorbant, eluent (in particular solvent mixtures), temperature and atmosphere
look directly at the UV source and wear make the application of Rf values to absolute identification rather problemat-
gloves if you put your hands into the UV ical. Usually an authentic sample is run along-side the unknowns in the mix-
cabinet. ture (Fig. 32.22) or on top of the mixture – ‘double spotting’ – as shown in
Fig. 32.23, to enable identification. The general procedure for running a TLC
plate is described in Box 32.3.
Box 32.3 How to run a TLC
1. Prepare the TLC development tank using a clean, dry spot. Do not allow the spot to be more than 2 mm in
beaker, as shown in Fig. 32.20, and allow it to equilib- diameter. Put the micropipette back into the sample
riate for 10 minutes. tube containing the mixture.
2. Prepare the plastic-backed TLC plate by drawing a 6. Spot other samples (e.g. reference compounds) onto
fine line (the base line) in pencil about 1 cm above the the plate as appropriate, using a new micropipette in
bottom. Take care not to scrape off any of the station- each case.
ary phase. Put three pencil dots on the base line – one
in the centre and the others equidistant on either side, 7. Evaporate the solvent from the plate by waving it in
but no closer than 3 mm to the edges of the plate. the air, holding the plate by the edges.
3. Dissolve the mixture (1–3 mg) in two or three drops of 8. Lower the plate into the developing tank, holding it
a suitable volatile solvent (dichloromethane or ether by the edges and make sure that the eluent does not
are the most common), in a small sample tube. cover the base line. If it does, discard the plate, pre-
pare another and empty some eluent from the tank.
4. Dip the tip of a micropipette into the solution and
capillary action will draw the solution into the pipette. 9. Put the lid on the tank and allow the eluent to rise up
the plate to about 1 cm from the top of the plate.
5. Touch the tip of the micropipette onto one of the pen-
cil spots on the base line. Capillary action will draw 10. Remove the chromatoplate from the tank and quickly
the solution from the micropipette onto the plate as a mark the height reached by the eluent, using a pencil.

Chromatography
11. Wave the chromatoplate in the air to evaporate the of solutions on the same spot.The result is non-separation
eluent from the plate. of the mixture and a ‘smear’ up the plate. Dilute the solu-
tion or don’t put so much on the plate.
12. Visualise the chromatogram by either placing the dry
chromatoplate in the UV cabinet and using a pencil Putting the spots of sample too close together. The sepa-
to draw round the spots (remember to wear gloves rated spots ‘bleed’ into each other and you can’t tell from
when your hands are in the UV cabinet) or putting the which sample they originate.
plate in the iodine jar until the dark spots develop.
Putting the spots too close to the edge of the plate. This
Problems with thin-layer chromatography results in inaccurate Rf values (spots travel faster up the
edge of the plate) and ‘bleeding’.
Overloading the chromatoplate with sample. TLC is an
extremely sensitive technique and it is easy to put too Contamination, which produces unexpected spots. Make
much sample on the plate. The sample solution must not sure all apparatus is clean, solvents are clean and use a
be too concentrated and you must not repeat applications fresh micropipette for each application.
Gravity chromatography
This is used for the preparative scale separation of mixtures of compounds,
where a sample of the mixture is separated by percolation down a column of
adsorbant by a suitable eluent. For effective separation the components of the
solvent
sand pressure mixture should have a difference in Rf value of at least 0.3. There are many
clamp
variations in detail of equipment and technique such as adsorbant, column
clamp packing (dry or slurry), sample application (including pre-adsorption on a small
sample of the stationary phase) and fraction collection, many of which are a
solvent
matter of personal choice based on personal experience. A typical arrangement
sand
is shown in Fig. 32.24(a) and for a detailed description of how to carry out
gravity chromatography you should consult specialist texts such as Errington
adsorbant adsorbant
(1997, p. 163), Harwood et al. (2000, p. 175) or Furniss et al. (1989, p. 209).
sinter Flash chromatography
clamp Flash chromatography or medium pressure chromatography has now almost
clamp completely replaced gravity chromatography for the preparative scale separa-
sand tion of mixtures. The use of smaller and more uniform particle size adsorbants
(alumina or silica) and the use of medium pressure via a nitrogen or air cylin-
der, a small air pump or even a pressure ball to pump the eluent through the
cotton wool
column gives more efficient (Rf difference of components down to 0.15) and
(a) (b)
much faster (minutes rather than hours) separations but at the expense of the
use of much greater volumes of eluent. A typical arrangement of equipment is
Fig. 32.24 Column chromatography: shown in Fig. 32.24(b), and Box 32.4 gives instructions on how to set up and
(a) gravity chromatography; (b) flash run a flash column based on our experiences. The references (p. 287) provide
chromatography. a more comprehensive insight into the methodology.
Key Points
1. Before attempting a preparative mixture separation by grav-
ity or flash chromatography, you must always analyse the
mixture by TLC to establish the stationary phase and eluent
parameters for effective separation and to determine the Rf
values of the components of the mixture.
2. You must analyse by TLC all the fractions collected from
the column to ensure that there is no ‘overlap’ of mixture
components.

Chromatography
3. Some chromatographers prefer to use dry packing for grav-

ity chromatography and slurry packing for flash chroma-
tography: the method used is often based on experience to
achieve the desired separation of components.
Box 32.4 How to prepare and run a flash column
Safety Notes
1. Flash alumina and silica are extremely fine and MUST be handled/transferred in the fume hood while you
are wearing a dust mask, since they are extremely harmful and tend to disperse in the air.
2. The column must be wrapped in plastic webbing or adhesive tape to prevent glass flying in case of explosion
when in use.
3. The pressure inlet must NOT be wired into place.
1. Prepare at least 30 clean and dry, numbered test silica and allow the column to cool. If eluent contin-
tubes in racks for fraction collection. Once you start ues to drip from the bottom of the column, with no
the chromatographic process you will not have time pressure, replace the amount being lost by addition
to prepare more. with the Pasteur pipette. AT NO STAGE SHOULD THE
ELUENT BE ALLOWED TO GO BELOW THE TOP OF THE
2. Clamp into a vertical position a Pyrex®, jointed chro- ADSORBANT/SAND.
matography column containing a No. 4 sinter and
mark the half height point. 10. Add more eluent to the column so that it is 75% full
and then add enough acid-washed dry sand, using
3. Prepare a fluted filter paper (see Box 24.1) and punc- the fluted filter paper technique, to produce a layer
ture a very small hole in the point. 1–2 cm deep on top of the silica.
4. Place the filter paper in a stemmed filter funnel in the 11. Pump the eluent down until it is about 1 cm above
top of the column and add the silica. the sand layer. The sand is there to prevent distur-
5. As the silica dribbles through into the column, tap bance to the flat top of the silica which will result in
the sides of the column with a piece of thick-walled uneven separation of the mixture.
rubber tubing to ensure even packing until the silica 12. Dissolve the mixture to be separated in the minimum
is about 2 cm below the half height mark. of eluent and add it to the eluent above the sand
6. Carefully add eluent to the top of the silica column, using the technique described in section 6.
either by using a long Pasteur pipette or by clamping a 13. Pump the eluent down to 1 cm above the sand, add
long glass rod down the centre of the column, almost another 5 mL of eluent and pump down again to 1 cm
but not touching the top of the adsorbant, and running above the sand. The mixture is now adsorbed on the
eluent down the glass rod. Try not to disturb the top of top of the column.
the stationary phase as you fill the column with eluent.
14. Fill the column with eluent (as in section 6) and pump
7. Rinse the top joint with a little eluent to wash any down to about 1 cm above the sand, collecting the
particles of adsorbant down on to the column. eluent from the bottom of the column as fraction 1,
or if you can see the components eluting, collect them
8. Switch on the air pump and hold the joint in the top
as separate fractions.
of the column as the eluent pumps down. The column
becomes opaque as the solvent passes through it and 15. Repeat the elution process until all the required frac-
becomes warm. tions have been collected.
9. Stop pumping, by removal of the pressure joint, 16. Run TLC on all the fractions collected to ensure that
when the eluent is about 2 cm from the top of the efficient separation has been effected.
Note: with the system described above, one pumped elution is equivalent to one ‘pass of solvent’ through the column.
This becomes important when a gradient elution (changing to eluent mixtures of increasing polarity) is required.

Chromatography
High-performance liquid chromatography

Safety Note Iodine vapour is toxic
and the iodine tank should be stored in High-performance liquid chromatography (HPLC) uses high pressure to force
the fume cupboard. the mobile phase through a closed column packed with micrometre-sized par-
ticles. This allows rapid separation of complex mixtures. Several operating
modes of HPLC are possible. These are:
UV visualisation – if you use an eluent ●● Normal phase (NPHPLC): the sample should be soluble in a hydrophobic
such as toluene, which absorbs in the solvent (e.g. hexane) and should be non-ionic. The mobile phase is non-polar
UV region, you must allow all the eluent while the stationary phase is polar (e.g. silica, cyano, amino).
to evaporate or you will see only a dark ●● Reversed phase (RPHPLC): the sample should be soluble in water or a
plate. polar organic solvent (e.g. methanol) and should be non-ionic. The mobile
phase is polar while the stationary phase is non-polar (e.g. C18 (ODS),
C8 (octyl), phenyl).
●● Size exclusion chromotography (SEC): this is used when the major dif-
ference between compounds in a mixture is their molecular weight. It is
normally used for compounds with molecular weights greater than 2000.
The mobile phase should be a strong solvent for the sample. Aqueous SEC
is called gel filtration chromatography (GFC) and is used for separation of
solvent sample
proteins and other biomolecules, while organic SEC is called gel permeation
reservoir injection detector chromatography (GPC) and is used for the separation of polymers.
valve
fraction
collector ●● Ion exchange chromotography (IEC): it is used when compounds
pump column
tube areionic, or potentially ionic (e.g. anions, cations, organic acids and bases,
amino acids, catecholamines, peptides). The mobile phase is typically a
buffer and the choice of pH is critical. Two types can be differentiated: SAX
solvent
(Strong-Anion eXchange) and SCX (Strong-Cation eXchange).
delivery microcomputer /
filter
tube readout The essential components of an HPLC system are a solvent delivery system, a
method of sample introduction, a column, a detector and an associated readout
Fig. 32.25 Components of an HPLC system. device (Fig. 32.25).
Solvent delivery system

This should fulfil certain requirements:
●● It should be chemically inert.
●● It should be capable of delivering a wide flow-rate range.
●● It should be able to withstand high pressures.
●● It should be able to deliver high flow-rate precision.
●● It should have a low internal volume.
●● It should provide minimum flow pulsation.
Although several systems are available that meet these requirements, the most
common is the reciprocating or piston pump. The choice of solvent delivery
system depends on the type of separation to be performed:
●● Isocratic separation: a single solvent (or solvent mixture) is used through-
out the analysis.
●● Gradient elution separation: the composition of the mobile phase is
altered using a microprocessor-controlled gradient programmer, which
mixes appropriate amounts of two different solvents to produce the required
gradient.

Chromatography
The main advantages of gradient HPLC are that you can control mobile-phase
HPLC is a versatile form of chroma-
composition. This allows you to resolve closely related compounds and pro-
tography, used with a wide variety of
vide faster elution of strongly retained compounds thereby producing reduced
stationary and mobile phases, to sepa-
analysis times and faster method development time. However, these advantages
rate individual compounds of a particu-
have to be compared with some disadvantages, such as the initial higher cost
lar class of molecules on the basis of
of the equipment compared with an isocratic system. Also, after each gradient
size, polarity, solubility or adsorption
run, a re-equilibration of the system is required to return to the initial mobile-
characteristics.
phase conditions.
Sample introduction
Preparing samples for HPLC – filter all The most common method of sample introduction in HPLC is via a rotary
samples through either a 0.2 mm or a valve (e.g. a Rheodyne® valve). A schematic diagram of a rotary valve is
0.45 mm filter prior to injection. shown in Fig. 32.26. In the load position, the sample is introduced via a
syringe to fill an external loop of volume 5, 10 or 20 mL. While this occurs,
the mobile phase passes through the valve to the column. In the inject posi-
tion, the valve is rotated so that the mobile phase is diverted through the sam-
sample in pump ple loop, thereby introducing a reproducible volume of the sample into the
1 mobile phase. The procedure for injection of a sample is shown in Fig. 32.27.
6 2 In Fig. 32.27(a) the syringe is filled with the sample/standard solution (typ-
loop ically 1 mL). Then the outside of the syringe is wiped clean with a tissue
5 3 (Fig. 32.27(b)). The syringe is placed into the Rheodyne ® injector of the
4
chromatograph while in the ‘load’ position (Fig. 32.27(c)) and the plunger
waste column on the syringe is depressed to fill the sample loop. Finally, the position of the
sample load Rheodyne valve is switched to the ‘inject’ position to introduce the sample
into the chromatograph (Fig. 32.27(d)) and then the syringe is removed from
pump the injection valve. The procedure for the preparation of a series of calibration
1
solutions is shown in Box 32.1.
6 2
5 3
4
column
sample inject
Fig. 32.26 Schematic diagram of a rotary

valve.
(a) (b)
(c) (d)
Fig. 32.27 Sample injection in HPLC.

Chromatography
The column
Always use the highest purity solvents.
This is usually made of stainless steel, and all components, valves, etc., are
manufactured from materials which can withstand the high pressures involved
Fig. 32.28. The most common form of liquid chromatography is reversed phase
HPLC. In RPHPLC, the most common column packing material consists of
C18 or octadecylsilane (ODS). A chemically bonded stationary phase is shown
in Fig. 32.29. However, some of the surface silanol groups remain unaffected.
These unreacted groups lead to undesirable chromatographic effects, such as
peak tailing (p. 253). One approach to remove the unreacted silanol groups is
end capping. In this way, the silanol group is reacted with a small silylating
group, e.g. trimethyl-chlorosilane. An alternative approach to nullify the action
of the silanol groups is to add triethylamine to the mobile phase, which modi-
fies the silica surface while in use.
Safety Note Always dispose of HPLC detectors

organic solvent waste in accordance
Most HPLC systems are linked to a continuous monitoring detector of high
with laboratory procedure (never down
sensitivity, for example phenols may be detected spectrophotometrically by
the sink).
monitoring the absorbance of the eluent at 280 nm as it passes through a flow
Mobile phase +
separated
sample out
Column
packing Stainless
Frit Column ferrule steel tubing
Column connection
Mobile phase
+ sample in
Fig. 32.28 Schematic diagram of the construction of an HPLC column.
Magnification ODS (C18) moiety on

surface of silica
CH3
H2
C
S
O H2C CH2
CH3
H2C CH2
H2C CH2
H2C CH2
Spherical silica
H 2C CH2
particle
H2 C CH2
CH2
Magnified silica H2C
CH2
surface showing H 2C
pore-like structure CH3
Fig. 32.29 A typical chemically bonded C18 stationary phase.

Chromatography
cell. Other detectors can be used to measure changes in fluorescence, current

Using silica-based HPLC columns –
or potential, as described below. Most detection systems are non-destructive,
these are limited to a pH range of 2–8
which means that you can collect eluent with an automatic fraction collector
(preferably 3–7). At low pH the bonded
for further study.
phase may be removed; at high pH the
UV/visible detectors (see also Chapter 29) are widely used and have the
silica particles may be dissolved.
advantages of versatility, sensitivity and stability. Such detectors are of two
types: fixed wavelength and variable wavelength. Fixed-wavelength detectors
are simple to use, with low operating costs. They usually contain a mercury
UV cutoff for organic solvents: lamp as a light source, emitting at several wavelengths between 254 nm and 578
nm; a particular wavelength is selected using suitable cutoff filters. The most
Hexane 195 nm
Acetonitrile 190 nm frequently used wavelengths for analysis of organic molecules are 254 nm and
Methanol 205 nm 280 nm. Variable wavelength detectors use a deuterium lamp and a continuously
Water 190 nm adjustable monochromator for wavelengths of 190–600 nm. For both types of
detector, sensitivity is in the absorbance range 0.001–1.0 (down to ≈1 ng),
with noise levels as low as 4 * 10-5. Note that sensitivity is partly influenced
from
by the path length of the flow cell, typically 10 mm (see Fig. 32.30). Monitor-
column ing at short-wavelength UV (e.g. below 240 nm) may give increased sensitivity
internal volume but decreased specificity, since many organic molecules absorb in this range.
typically 5–10 µL Additional problems with short-wavelength UV detection include instrument
instability, giving a variable base line, and absorption by components of the
mobile phase (e.g. organic solvents, which often absorb at 6 210 nm).
An important development in chromatographic monitoring is diode array
detection (DAD). The incident light comprises the whole spectrum of light from
the source, which is passed through a diffraction grating and the diffracted light
quartz quartz
window window detected by an array of photodiodes. Typical DAD can measure the absorbance
of each sample component at 1–10 nm intervals over the range 190–600 nm.
This gives an absorbance spectrum for each eluting substance which may be
used to identify the compound and give some indication as to its purity. An
example of a three-dimensional diode array spectrum is shown in Fig. 32.31.
Many aromatic organic molecules, including some polycyclic aromatic
hydrocarbons, show natural fluorescence, or can be made to fluoresce by
path length pre-column or post-column derivatisation with a fluorophore. Fluorescence
typically 10 mm detection is more sensitive than UV/visible detection, and may allow analysis in
the picogram (10-12 g) range. A fluorescence detector consists of a light source
Fig. 32.30 UV detector cell for HPLC. (e.g. a xenon lamp), a diffraction grating to supply light at the excitation wave-
length, and a photomultiplier to monitor the emitted light (usually arranged to
be at right angles to the excitation beam). The use of instruments with a laser
light source can give an extremely narrow excitation waveband, and increased
sensitivity and specificity.
Electrochemical detectors offer very high sensitivity and specificity, with
the possibility of detection of femtogram amounts of electroactive compounds
such as catecholamines, vitamins, thiols, purines, ascorbate and uric acid. The
two main types of detector, amperometric and coulometric, operate on simi-
lar principles, i.e. by measuring the change in current or potential as sample
components pass between two electrodes within the flow cell. One of these
electrodes acts as a reference (or counter) electrode (e.g. calomel electrode),
while the other – the working electrode – is held at a voltage that is high enough
to cause either oxidation or reduction of sample molecules. In the oxidative
Overcoming interference with fluores- mode, the working electrode is usually glassy carbon, while in reductive mode
cence detectors – use a dual flow cell a mercury electrode is used. In either case, a current flow between the elec-
to offset background fluorescence due trodes is induced and detected.
to components of the mobile phase. Mass spectrometry (p. 356) used in conjunction with chromatographic
methods can provide a powerful tool for identifying the components of complex

Chromatography
mixtures (e.g. pharmaceuticals). One drawback is the limited capacity of the

mass spectrometer – due to its vacuum requirements – compared with the
volume of material leaving the chromatography column. Similarly, in HPLC,
devices have been developed for solving the problem of large solvent volumes,
absorbance
for example by splitting the eluent from the column so only a small fraction
reaches the mass spectrometer.
The computer-generated outputs from the mass spectrometer are similar to
chromatograms obtained from other methods, and show peaks corresponding
to the elution of particular components. However, it is then possible to select
an individual peak and obtain a mass spectrum for the component in that peak
to aid in its identification (p. 363). This has helped to identify hundreds of
(nm
) components present in a single sample, including flavour molecules in food,
drug metabolites and water pollutants.
gth
elen
wav
Recording and interpreting chromatograms

time (min)
For analytical purposes, the detector output is usually connected to a comput-
Fig. 32.31 Diode array detector absorption er-based data acquisition and analysis system. This consists of a personal com-
spectra of the eluent from an HPLC sepa- puter (PC) with data acquisition hardware to convert an analogue detector signal
ration of a mixture of four steroids, taken to digital format, plus software to control the data acquisition process, store the
every 15 seconds. signal information and display the resulting chromatogram. The software will
also detect peaks and calculate their retention times and sizes (areas) for quanti-
tative analysis. The software often incorporates functions to control the chroma-
Maximising sensitivity with fluores- tographic equipment, enabling automatic operation. In sophisticated systems, the
cence detectors – the concentration detector output may be compared with that from a ‘library’ of chromatograms
of other sample components, e.g. pig- for known compounds, to suggest possible identities of unknown sample peaks.
ments, must not be so high that they In simpler chromatographic systems, you may need to use a chart recorder
cause quenching of fluorescence. for detector output. Two important settings must be considered before using a
chart recorder:
1. The base-line reading – this should be set only after a suitable quantity
Optimising electrochemical detection – of mobile phase has passed through the column (prior to injection of the
the mobile phase must be free of any sample) and stability is established. The chart recorder is usually set a little
compounds that might give a response; above the edge of the chart paper grid, to allow for base-line drift.
all constituents must be of the highest 2. The detector range – this must be set to ensure that the largest peaks do
purity. not go off the top of the chart. Adjustment may be based on the expected
quantity of analyte, or by a trial-and-error process. Use the maximum sen-
sitivity that gives intact peaks. If peaks are still too large on the minimum
sensitivity, you may need to reduce the amount of sample used, or prepare
Interpreting chromatograms – never
and analyse a diluted sample.
assume that a single peak is a guaran-
tee of purity: there may be more than
one compound with the same chroma- Interpreting chromatograms
tographic characteristics.
Make sure you know the direction of the horizontal axis of the chromatogram
(usually, either volume or time) – it may run from right to left or vice versa –
and make a note of the detector sensitivity on the vertical axis. Ideally, the
Problems with peaks – non-symmetrical base line should be ‘flat’ between peaks, but it may drift up or down owing to
peaks may result from column overload- a number of factors including:
ing, co-elution of solutes, poor packing ●● changes in the composition of the mobile phase (e.g. in gradient elution);
of the stationary phase, or interactions
between the substance and the support ●● tailing of material from previous peaks;
material. ●● carry-over of material from previous samples; this can be avoided by effi-
cient cleaning of columns between runs – allow sufficient time for the

Chromatography
previous sample to pass through the column before you introduce the next
Avoiding problems with air bubbles
sample;
in liquid chromatography – always
ensure that buffers are effectively ●● loss of the stationary phase from the column (column ‘bleed’), caused by
degassed by vacuum treatment before extreme elution conditions;
use, and regularly clean the flow cell of
●● air bubbles (in liquid chromatography); if the buffers used in the mobile
the detector.
phase are not effectively degassed, air bubbles may build up in the flow cell
of the detector, leading to a gradual upward drift of the base line, followed
by a sharp fall when the accumulated air is released. Small air bubbles that
Degassing your mobile phase solvent – do not become trapped may give spurious small peaks as they pass through
this is an important step and the best the detector.
approach is to prepare the solvent com- A peak close to the origin may be due to non-retained sample molecules, flow-
position (e.g. 50 : 50 v/v methanol : ing at the same rate as the mobile phase, or to artefacts, for example, air (GC)
water, for isocratic RPHPLC) and then or solvent (HPLC) in the sample. Whatever its origin, this peak can be used to
filter through a 0.22 mm porosity filter measure the void volume and dead time of the column (p. 252). No peaks from
using a Büchner flask arrangement. genuine sample components should appear before this type of peak.
Peaks can be denoted on the basis of their elution volume (used mainly in
liquid chromatography) or their retention times (mainly in GC). If the peaks
are not narrow and symmetrical, they may contain more than one component.
Where peaks are more curved on the trailing side compared with the leading
side (peak tailing, p. 253), this may indicate too great an association between
the component and the stationary phase, or overloading of the column.
Optimising chromatographic separations

In an ideal chromatographic analysis, the sample molecules will be completely
separated, and detection of components will result in a series of discrete indi-
vidual peaks corresponding to each type of molecule. However, to minimise
the possibility of overlapping peaks, or of peaks composed of more than one
substance, it is important to maximise the separation efficiency of the tech-
nique, which depends on:
●● the selectivity, as measured by the relative retention times of the two com-
ponents (p. 252), or by the volume of the mobile phase between the peak
maxima of the two components after they have passed through the column;
this depends on the ability of the chromatographic method to separate two
components with similar properties;
●● the band-broadening properties of the chromatographic system, which influ-
ence the width of the peaks; these are mainly due to the effects of diffusion.
The resolution of two adjacent components can be defined in terms of k′, a
and N, using eqn [32.8].
Key Point In practical terms, good resolution is achieved

when there is a large ‘distance’ (either time or volume) between
peak maxima, and the peaks are as narrow as possible.
The resolution of components is also affected by the relative amount of each

substance: for systems showing low resolution, it can be difficult to resolve
small amounts of a particular component in the presence of larger amounts of
a second component. If you cannot obtain the desired results from a poorly

Chromatography
resolved chromatogram, other chromatographic conditions, or even different

Quantifying molecules – note that
methods, should be tried in an attempt to improve resolution. For liquid chro-
quantitative analysis often requires
matography, changes in the following factors may improve resolution:
assumptions about the identity of sep-
arated components and that further ●● Stationary-phase particle size – the smaller the particle, the greater the area
techniques may be required to provide available for partitioning between the mobile phase and the stationary phase.
information about the nature of the This partly accounts for the high resolution observed with HPLC compared
molecules present, e.g. mass spectrom- with low-pressure methods.
etry (see Chapter 38).
●● The slope of the salt gradient in eluting IEC columns (e.g. using comput-
er-controlled adapted gradients).
●● In low-pressure liquid chromatography, the flow rate of the mobile phase
must be optimised because this influences two band-broadening effects
which are dependent on diffusion of sample molecules: (i) the flow rate must
be slow enough to allow effective partitioning between the mobile phase and
the stationary phase: and (ii) it must be fast enough to ensure that there is
minimal diffusion along the column once the molecules have been separated.
To allow for these opposing influences, a compromise flow rate must be used.
●● If you prepare your own columns, they must be packed correctly, with no
channels present that might result in uneven flow and eddy diffusion.
When using external standardisation – Quantitative analysis

samples and standards should be ana- Most detectors and chemical assay systems give a linear response with increas-
lysed more than once, to confirm the ing amounts of the test substance over a given ‘working range’. Alternative
reproducibility of the technique. ways of converting the measured response to an amount of substance are:
When using an internal standard,
you should add an internal standard ●● External standardisation: this is applicable where the sample volume is suf-
to the sample at the first stage in the ficiently precise to give reproducible results (e.g. HPLC). You measure the
extraction procedure, so that any loss peak areas (or heights) of known amounts of the substance to give a calibra-
or degradation of test substancedur- tion factor or calibration curve (Chapter 49) which can be used to calculate
ing purification is accompanied by an the amount of test substance in the sample.
equivalent change in the internal stand- ●● Internal standardisation: where you add a known amount of a reference sub-
ard, as long as the extraction character- stance (not originally present in the sample) to the sample, to give an addi-
istics of the internal standard and the tional peak in the elution profile. You determine the response of the detector
test substance are very similar. to the test and reference substances by analysing a standard containing
known amounts of both substances, to provide a response factor (r), where:
peak area (or height) of test substance
r = [32.12]
peak area (or height) of reference substance
Use this response factor to quantify the amount of test substance (Qt) in a
sample containing a known amount of the reference substance (Qr), from the
relationship:
[peak area (or height) of test substance] Qr
Qt = * [32.13]
[peak area (or height) of reference substance] r
Internal standardisation should be the method of choice wherever possible, since

Internal standard – The internal stand-
it is unaffected by small variations in sample volume (e.g. for GC microsyringe
ard should be chemically similar to the
injection). An additional advantage of an internal standard which is chemically
test substance(s) and must give a peak
related to the test substance is that it may show up problems due to changes in
that is distinct from all other substances
in the sample.
detector response, incomplete derivatisation, etc. A disadvantage is that it may
be difficult to fit an internal standard peak into a complex chromatogram.

Chromatography
Sample preparation for chromatography

The preparation of a sample for subsequent chromatographic analysis is critical
to the chemist. Often the key steps in the sample preparation are to ensure that
the sample acquired is:
●● not mislaid or lost,
●● converted to a form that is suitable for subsequent chromatographic analysis;
●● not contaminated.
Key Point Sample preparation is also used for removal

of target analyte(s) from a sample matrix, and for their pre-
concentration prior to chromatographic analysis.
A range of analytical procedures can be used depending upon whether the

sample can be classified as either solid (or semi-solid), an aqueous solution or
a gas (air) sample. In either situation, a range of sample preparation techniques
can be used.
The apparatus and procedure for Soxhlet extraction is shown in Fig. 32.32.
A more modern version of Soxhlet extraction is Soxtec extraction. This has the
advantages of:
●● rapid extraction (approximately 2 hrs per sample);
●● less organic solvent usage (upto 80% less than Soxhlet);
●● sample extract can be concentrated in situ.
(a) (b) (c) (d) (e)
Fig. 32.32 Apparatus and procedure for Soxhlet extraction: (a) experimental set-up for Soxhlet extraction, (b) sample
placed in thimble inside Soxhlet, (c) heat applied to organic solvent, (d) organic solvent vapour condenses through the
sample (in the thimble), and, (e) solvent-containing extracted organic compounds returns to round-bottomed flask.

Chromatography
(a) (b) (c)
Boiling: Rapid Rinsing: Efficient Recovery: Automatic

solubilisation in removal of remaining collection of distilled
boiling solvent soluble matter solvent for re-use
Fig. 32.33 Apparatus and procedure for Soxtec extraction.
A schematic diagram of the apparatus and procedure for Soxtec extraction is

Sample preparation techniques for
shown in Fig. 32.33. Box 32.5 outlines the procedure for Soxtec extraction.
solid samples
In shake flask extraction, the sample (accurately weighted) is placed in
●● Soxhlet extraction a suitable stoppesed glass container and organic soluent added. Agitation via
●● Soxtec extraction either hand or mechanical device then takes place. Fig. 32.34 shows the most
●● shake flask extraction
effective extraction process via end-over-end action. The procedure for shake
●● utrasonic extraction
●● supercritical fluid extraction
flask extraction is shown in Box 32.6.
●● microwave-assisted extraction Ultrasonic extraction (or sonication) Fig. 32.35 uses sound waves to agi-
●● pressurised fluid extraction. gate a sample immesed in organic solvent. This can be done using either a sonic
probe (Fig. 32.35(a)) or an ultrasonic bath (Fig. 32.35 (b)). Box 32.7 highlights
Supercritical CO2 occurs at 7 31.1 ° C,
the procedure for ultrasonic extraction.
and 74.8 atm
Microwave radiation is produced by the Preparation of solid (or semi-solid) samples
magnetion.
The most common approach for recovery of analytes from solid (or semi-solid)
samples is Soxhlet extraction (see p. 143 and Fig. 15.5). Box 15.2 In addition,
several modern techniques can be used, including supercritical fluid extraction,
Box 32.5 How to use a Soxtec extractor
1. Accurately weigh sample (e.g. 5 g) and anhydrous 3. Thimble (containing the sample is then elevated above
sodium sulphate (e.g. 5 g) into cellulose extraction the boiling solvent (Fig. 32.33(b) for upto 60 mins.
thimble.
4. Solvent evaportaion then takes place (Fig. 32.33(c))
for 10–15 mins.
2. Immerse the sample containing thimble into the boil-
ing solvent (e.g. 60 mL of acetone: dichloromethane, 5. The final concentrated extract is quantitatively trans-
1:1, v/v for 60 mins (Fig. 32.33(a)). ferred to a volumetric flask ready for analysis.

Chromatography
Fig. 32.34 Apparatus for shake flask extraction.
(a) microwave-assisted extraction and pressurised fluid extraction (also known by

Sonic probe: control box
its trade name of accelerated solvent extraction). These instrumental techniques
rely on the use of heat, pressure and solvents to extract organic compounds
Sonic probe: inserted in container from solid or semi-solid samples. Supercritical fluid extraction (SFE) exploits
containing sample and solvent
the gas-like and liquid-like properties of a supercritical fluid, typically CO2,
to extract organic compounds at temperatures above 31.1°C at a pressure of
(b) 74.8 atm (1070.4 psi). By using combinations of CO2 mixed with an organic
Sample placed inside container
within the sonic bath
modifier (e.g. methanol), it is possible to extract a range of organic molecules of
different polarity. Instrumentally, the system consists of a source of CO2, which
is pumped (after cooling of the pump head) to the extraction cell. A second
pump can be added for the organic modifier. The extraction cell is located in
an oven while pressure is generated in the system via a back-pressure regulator
(Fig. 32.36). Typically, samples are extracted for 10–60 min, depending on
Fig. 32.35 Ultrasonic extraction using
either (a) a sonic probe or (b) an ultrasonic
the organic compounds present and the temperature and pressure conditions
bath. selected, and collected ready for analysis. A procedure for the SFE of organic
compounds is described in Box 32.8.
In pressurised microwave-assisted extraction (MAE), an organic solvent
and the sample are subjected to radiation from the microwave source (i.e. the
Box 32.6 Procedure for Shake Flask Extraction
1. An accurately weighted sample (e.g. 1 g) is placed 4. The extract-containing solvent is then removed and
in a suitable stoppered container. stored.
2. Organic solvent is added (e.g. 10 mL) of acetone: 5. Fresh organic solvent is added and steps 3–4 are
dichloromethane, 1:1, v/v). repeated a further two times.
3. After securing the stopper the sample is agitated 6. The combined extracts can either be analysed
by rotating on an end-over-end shaker (Fig. 32.34) for directly or the solvent evaporated (see p. 162) prior
30 mins. to analysis.

Chromatography
CO2 magnetron) in a sealed vessel (Fig. 32.37). The sample and solvent are placed
supply oven pressure
control into an inert vessel liner (100 mL) made from a fluoropolymer and subject to
extraction heating for a period of time (thereby resulting in the build up of pressure). In
pump cell vial
the event of an extraction cell reaching critical conditions, an automatic system
00.0
allows venting of excess pressure. If any solvent leaks from an extraction vessel
a solvent monitoring system will automatically shut off the magnetron but leave
the exhaust fan to continue working. Most systems allow up to 14 samples to be
Fig. 32.36 Schematic diagram of a super-
extracted simultaneously. A procedure for the extraction of organic compounds
critical fluid extraction system.
by MAE is described in Box 32.9.
Pressurised fluid extraction, available commercially as accelerated sol-
vent extraction, ASE™, is an automated system capable of processing up to
24 samples sequentially (Fig. 32.38). Each sample is placed in an extraction
cell and loaded on a carousel. After setting the extraction conditions, typically
a temperature of 100 °C and a pressure of 2000 psi for an extraction time of
10 min, organic solvent is introduced into the extraction cell. Upon completion
the extract is collected in a vial (a flow of nitrogen gas is used to remove trace
amounts of solvent) and analysed. A procedure for the extraction of organic
compounds by ASE™ is described in Box 32.10.
Box 32.7 Procedure for ultrasonic extraction
1. An accurately weighted sample (e.g. 1 g) is placed in 4. The extract-containing solvent is then removed and
a suitable glass container. stored.
2. Organic solvent is added (e.g. 10 ml of acetone: 5. Fresh organic solvent is added and steps 3-4 are
dichloromethane, 1:1, v/v). repeated a for then two times.
3. The sonic probe is placed in the solvent-sample mix- 6. The combined extracts can either be analysed directly
ture and switched on (e.g. for 3 mins). or the solvent evaporated (p. 162) prior to analysis.
Box 32.8 How to operate a typical supercritical fluid extraction system
1. Turn on the electrical supply of the SFE system 6. Set the SFE operating parameters: flow rate of liquid
including recirculating water bath. Allow 30 mins for CO2, 2 mL min -1 and methanol, 0.2 mL min -1; oven
cooling of the CO2 pump head. temperature, 60 °C; and, pressure, 250 kg cm2. Before
the extraction commences, pre-heat the extraction
2. Take an extraction cell and tighten an end cap on one
cell containing the sample to the pre-set tempera-
end only using a wrench and then weigh the cell.
ture for 10 min, then undergo a static extraction (no
Ensure the extraction cell is suitable for its purpose,
flow of CO2) at the operating conditions for 5 min and
i.e. able to withstand high pressure and does not leak.
finally a dynamic extraction (flow of CO2 and metha-
3. Fill the extraction cell with the sample mixed 50 : 50 nol) for one hour.
with an inert matrix (e.g. Celite™), and weigh the cell
7. Remove the collection vial from the system after the
again.
allotted extraction time, and back-flush the C18 SPE
4. Tighten the other end cap onto the cell with the cartridge with fresh methanol (2 mL).
wrench and insert into the oven of the SFE system.
8. Carefully transfer extract to a volumetric flask. Ensure
This requires the use of a wrench to ensure a suitable
all extract is transferred by washing the collection vial
connection.
with small quantities of solvent.
5. Connect a glass collection vial containing 2 mL of
9. Analyse the extract using an appropriate technique,
methanol to the outlet of the back-pressure regulator
for example, gas chromatography.
fitted with a C18 SPE cartridge (p. 280).

Chromatography
microwave Preparation of aqueous samples

apparatus
Centrifugation
vent One of the simplest forms of sample preparation for aqueous samples contain-
tube 00.0
ing particulates is centrifugation.
Low-speed centrifuges
sample Bench-top instruments for routine use, have a maximum speed of 3000–
vessels
6000 r.p.m. and a relative centrifugal field (RCF, or ‘g value’) up to 6000 g.
Most modern machines also have a sensor that detects any imbalance when
the rotor is spinning and cuts off the power supply. Box 32.11 gives details of
operation for a low-speed centrifuge.
Centrifuges can be used with sample tubes of different size and capacity by
Fig. 32.37 Schematic diagram of a pressur- changing the rotor. In swing-out rotors, the sample tubes are placed in buckets
ised microwave-assisted extraction system. which pivot as the rotor accelerates (Fig. 32.39). Swing-out rotors are used on
Box 32.9 How to operate a typical pressurised microwave-assisted extraction (MAE) system
1. Take an extraction cell and then weigh the cell. the extraction, while temperature is monitored for all
cells every 7 s.
2. Fill each extraction cell with sample, approximately
2 g, and weigh the cell again. 6. After the allotted extraction time, remove all the
extraction cells on the carousel and allow to cool
3. Add 20–50 mL of organic solvent to each sample.
for approximately 30 min before attempting to open
Typical solvents include dichloromethane, acetone
them.
and methanol. Non-polar solvents (e.g. hexane) can
be used by mixing them with polar solvents. 7. Filter each extract to remove the sample matrix from
the organic solvent containing solvent.
4. Tighten the end cap on to the cell and insert into the
microwave oven of the MAE system. All extraction 8. Carefully transfer extract to a volumetric flask. Ensure
cells are mounted in a carousel. that all extract is transferred by washing the collection
vial with small quantities of solvent.
5. Set the MAE operating parameters: temperature,
200 °C; pressure, 250 psi; extraction time, 10 min. 9. Analyse the extract using an appropriate technique,
Typically, pressure is continuously measured during for example, gas chromatography.
pump position path length

valve
during
centrifugation
valve oven
vent extraction
cell
nitrogen
cylinder vent position at rest rmin
solvent
collection rav
vial
rmax
Fig. 32.39 Swing-out rotor

Fig. 32.38 Schematic diagram of an
accelerated solvent extraction system.

Chromatography
Box 32.10 How to operate a typical pressurised fluid extraction (PFE) system
1. Ensure the system is connected to the electrical 5. Typical PFE operating parameters are temperature,
supply and is ready for operation with a nitrogen 100 °C; pressure, 2000 psi; extraction time, 10 min.
supply and organic solvent. Ensure collection vials
6. After the allotted extraction time, remove the extract
are in place.
containing collection vial.
2. Take an extraction cell and finger tighten an end cap
on one end only and then weigh the cell.
that all extract is transferred by washing the collection
3. Fill the extraction cell with the sample, 2–10 g, mixed vial with small quantities of solvent.
50 : 50 with an inert matrix (e.g. Celite™) and weigh
the cell again.
for example, gas chromatography.
4. Finger tighten the other end cap on to the cell and
place in carousel and start the extraction programme.
Box 32.11 How to use a low-speed bench centrifuge
1. Choose the appropriate tube size and material for 5. For centrifuges with swing-out rotors, check that
your application, with caps where necessary. Most each holder/bucket is correctly positioned in its locat-
low-speed machines have four-place or six-place ing slots on the rotor and that it is able to swing freely.
rotors – use the correct number of samples to fill the All buckets must be in position on a swing-out rotor,
rotor assembly whenever possible. even if they do not contain sample tubes - buckets are
an integral part of the rotor assembly.
2. Fill the containers to the appropriate level: do
not overfill, or the sample may spill during 6. Load the sample tubes into the centrifuge. Make sure
centrifugation. that the outside of the centrifuge tubes, the sample
holders and sample chambers are dry: any liquid pres-
3. It is vital that the rotor is balanced during use. There-
ent will cause an imbalance during centrifugation, in
fore, identical tubes must be prepared, to be placed
addition to the corrosive damage it may cause to the
opposite each other in the rotor assembly. This is
rotor. For sample holders where rubber cushions are
particularly important for density gradient samples,
provided, make sure that these are correctly located.
or for samples containing materials of widely dif-
Balanced tubes must be placed opposite each other –
fering densities (e.g. soil samples) since the density
use a simple code if necessary, to prevent mix-ups.
profile of the tube will change during a run. How-
ever, for low-speed work using small amounts of par- 7. Bring the centrifuge up to operating speed by gentle
ticulate matter in aqueous solution, it is sufficient to acceleration. Do not exceed the maximum speed for
counterbalance a sample with a second tube filled the rotor and tubes used.
with water, or a saline solution of similar density to
8. If the centrifuge vibrates at any time during use,
the sample.
switch off and find the source of the problem.
4. Balance each pair of sample tubes (plus the cor-
9. Once the rotor has stopped spinning, release the lid
responding caps, where necessary) to within 0.1 g
and remove all tubes. If any sample has spilled, make
using a top-pan balance; add liquid dropwise to the
sure you clean it up thoroughly using a non-corrosive
lighter tube, until the desired weight is reached. Alter-
disinfectant (e.g. Virkon®) so that it is ready for the
natively, use a set of scales. For small sample vol-
next user.
umes (up to 10 mL) added to disposable, lightweight
plastic tubes, accurate pipetting of your solution may 10. Close the lid (to prevent the entry of dust) and return
be sufficient for low-speed use. all controls to zero.

Chromatography
low-speed centrifuges: their major drawback is their extended path length and
Safe working practice with centrifuge
the resuspension of pellets due to currents created during deceleration.
tubes – never be tempted to use a tube
or bottle which was not designed to fit
the machine you are using (e.g. a gen-
Key Point It is vital that you balance your loaded centrifuge
eral-purpose glass test-tube, or a screw-
tubes before use. As a general rule, balance all sample tubes
capped bottle), or you may damage the
to within 1% or better, using a top-pan balance or scales. Place
centrifuge and cause an accident.
balanced tubes opposite each other.
Balancing the rotor – For the safe use Solvent extraction

of centrifuges, the rotor must be bal-
anced during use, or the spindle and The most common approaches for preparing aqueous samples for chromato-
rotor assembly may be damaged per- graphic analysis involve solvent extraction. Typically, this is done using either
manently; in severe cases, the rotor liquid–liquid extraction (Chapter 15) or solid–liquid extraction. The most com-
may fail and cause a serious accident. mon form of solid–liquid extraction is solid phase extraction.
Solid phase extraction
Solid phase extraction (SPE) is a technique that can be used for clean-up and
Using microcentrifuge tubes – the inte-
pre-concentration of aqueous samples. The aqueous sample is passed through
gral push-on caps of microcentrifuge
a sorbent (typically the same material as the stationary phase in HPLC, see
tubes must be correctly pushed home
p. 267) via gravity or, more likely, with the aid of a vacuum. The sorbent is
before use or they may come off during
usually packed in to small tubes or cartridges (Fig. 32.40). The primary func-
centrifugation.
tion of the sorbent is to retain the analyte(s) in the aqueous sample, but not
to retain any extraneous material present. After this pre-concentration and/
or clean-up step the analyte(s) are eluted from the sorbent and collected for
Safe practice – Given their speed of subsequent analysis.
rotation and the extremely high forces
generated, centrifuges have the poten-
tial to be extremely dangerous, if used
SPE cartridge
incorrectly. The most common arrangement for SPE is the syringe barrel or cartridge. The
cartridge itself is usually made of polypropylene with a wide entrance, through
which the sample is introduced, and a narrow exit (male luer tip). The sorbent
Balancing tubes – never balance cen- material, ranging in mass from 50 mg to 10 g, is positioned between two frits,
trifuge tubes ‘by eye’ – use a balance. at the base (exit) of the cartridge and on top of the sorbent, which act to both
Note that a 35 mL tube full of liquid at retain the sorbent material and to filter out any particulate matter (Fig. 32.40).
an RCF of 3000 g has an effective weight Solvent flow through a single cartridge is typically done using a side-arm
greater than a large adult man. flask apparatus (Fig. 32.41), whereas multiple cartridges can be simultaneously
processed (from 8 to 30 cartridges) using a vacuum manifold (Fig. 32.42).
Recent advances in automation have allowed modified autosamplers (devices
For safety reasons, all centrifuges are for sample manipulation) to be used to provide unattended operation of an SPE
manufactured with an armoured casing
that should contain any fragments in
cases of rotor failure. Machines usually polypropylene body
have a safety lock to prevent the motor
from being switched on unless the lid
fritted disk
is closed and to stop the lid from being
opened while the rotor is moving. sorbent bed
fritted disk
luer tip
SPF cartridges – these contain a range
of sorberts, classified as normal phase,
reversed phase or ion-exchange.
Fig. 32.40 Solid phase extraction cartridge.

Chromatography
sorbent bed sorbent bed
bung
sample
collection vial
vacuum
applied
vacuum
applied
Fig. 32.42 Manifold for multiple solid phase extraction cartridges.
Fig. 32.41 Manifold for a single solid phase

extraction cartridge.
system with the potential to link the system to the analysis stage. The use of
automated SPE allows more samples to be extracted (higher sample through-
put) with better precision. In addition, it also allows the scientist to perform
other tasks or prepare more samples for analysis.
Types of SPE media

Sorbents for SPE can be classified as normal phase, reversed phase or ion
Reversed phase sorbents – these use
exchange. The most common sorbents are based on silica particles (see p. 267)
non-polar sorbents and polar solvents
to which functional groups are bonded to surface silanol groups to alter their
while ’normal’ phase sorbents use polar
retentive properties. In addition to silica, some other common sorbents are
sorbents and non-polar solvents.
based on florisil, alumina and macroreticular polymers.
Normal phase sorbents have polar functional groups (e.g. cyano, amino
and diol groups) and hence can retain polar analytes (e.g. drugs, phenols). In
contrast, reversed phase sorbents have non-polar functional groups (e.g. octa-
decyl, octyl and methyl groups) and hence can retain non-polar compounds
(e.g. polycyclic aromatic hydrocarbons). Ion-exchange sorbents have either
cationic or anionic functional groups that attract compounds of the opposite
charge in the ionised form. For example, a cation-exchange phase, such as
benzenesulfonic acid, will extract an analyte with a net positive charge (e.g. a
phenoxyacid herbicide).
Method of SPE operation

Careful optimisation of the SPF method The mode of operation of SPE can be divided in to five steps (Box 32.12).
is required if the approach is to be Each step is characterised by the nature and type of solvent used which in turn
succesful. is dependent upon the characteristics of the sorbent and the sample. The five
steps are:
Solvent evaporation may be required 1. Wetting the sorbent
post-extraction to achieve the greatest 2. Conditioning the sorbent
sensitivity.
3. Loading the sample

Chromatography
Box 32.12 H
ow to pre-concentrate a sample using a reversed phase C18 solid phase
extraction (SPE) cartridge
1. Place the SPE cartridge into a vacuum manifold. 5. Remove unwanted, extraneous material by washing
Apply the vacuum at a flow rate of 5 mL min -1. the sample-containing sorbent with a high-polarity
solvent or buffer. This process may be repeated.
2. Wet the sorbent by passing 1.0 mL of methanol or
6. Elute analytes from the cartridge with a less polar
acetonitrile through the cartridge. This solvent will
solvent, for example methanol or the HPLC mobile
remove impurities from the C18 sorbent and wet its
phase (if this is the method of subsequent analysis).
surface.
3. Condition the sorbent by passing 1 mL of water or that all extract is transferred by washing the collection
buffer through the sorbent. Do not allow the sorbent vial with small quantities of solvent.
to dry out before applying the sample.
for example gas chromatography.
4. Add the sample to the sorbent in a high-polarity sol-
vent or buffer. 9. Discard the SPE cartridge.
4. Rinsing or washing the sorbent to elute extraneous material

5. Eluting and collecting the analyte of interest.
The choice of solvents for SPE are obviously important in the overall success
of this method. Further guidance on solvent selection is found on p. 139. In
practice, at the wetting stage a solvent is required that will ‘activate’ the sorb-
ent whereas the conditioning sorbent is required to prepare the sorbent for
the aqueous sample. Perhaps the most difficult solvent to select is the rinsing
or washing solvent. In this step the function of the solvent is to remove any
mechanism for extraneous material but not the analyte(s) of interest. Finally, the choice of
raising/lowering elution solvent is one that can both effectively remove the analyte(s) from the
of fibre
sorbent and can also be compatible with the method of analysis to be used. If
the eluting solvent proves to be incompatible with the analytical technique of
choice it is possible to carry out a solvent exchange. This can be done by evap-
orating the analyte(s) containing solvent to dryness, using a solvent evaporation
technique (p. 162), and then to reconstitute the analyte(s) in a different solvent.
This process can be problematical in that volatile analyte(s) can be lost due to
evaporation: it is wise to evaluate the approach on a test sample first.
Solid phase microextraction (SPME)

Solid phase microextraction is a relatively new technique for the preparation of
samples for chromatography. It uses a coated fused-silica fibre to retain organic
stainless compounds, i.e. to pre-concentrate them from the sample matrix, for example
steel needle air, aqueous or solid. Desorption of organic compounds normally occurs in the
hot injection port of a gas chromatograph (GC), although it is possible to use
the mobile phase of a high-performance liquid chromatography (HPLC) system
support to desorb the organic compounds.
for fibre
The SPME device consists of a fused silica fibre, coated with a stationary
phase and mounted in a syringe-type holder (Fig. 32.43). The holder has two
coated
silica fibre
functions:
1. to provide protection for the fibre
Fig. 32.43 Solid phase microextraction 2. to allow insertion into the hot environment of the GC injection port (or
system.
injection valve of an HPLC) using a needle.

Chromatography
(b) As samples and standards are normally introduced into chromatographic sys-
(a) tems using a syringe (p. 257) the use of an SPME device is straight-forward.
SPME fibre When not in use the fused-silica coated fibre is retracted within the pro-
SPME fibre
holder tective needle of the holder. In operation, the fibre (∼1 cm long) is exposed to
holder organic compounds within their matrix (air, aqueous, solid) for a specified time.
SPME can be used in two ways (Fig. 32.44):
1. direct
cap with cap with
septum septum 2. headspace SPME.
coated fibre
vial In direct SPME, the coated silica fibre is exposed to the sample matrix directly
vial
coated fibre (e.g. blood plasma). A procedure for the analysis of an organic compound in
sample
aqueous solution by SPME is shown in Box 32.13. Improved response time and
Fig. 32.44 Schematic diagrams repre-sent- signal can be achieved by agitation of the sample vial, agitation of the fibre,
ing the two approaches in which SPME can
and/or stirring or sonication of the sample solution. For gaseous samples, the
be used (a) direct SPME involving insertion
natural convection of air is usually sufficient to aid diffusion of analyte(s) onto
of the fibre directly into the aqueous sam-
ple and (b) headspace SPME where the the SPME fibre. In headspace SPME, the process relies on the release of volatile
fibre is suspended above the sample. compounds from the sample matrix. This may be achieved by heat, chemical
modification or the inherent volatility of the organic compounds. A procedure
for the analysis of an organic compound in the headspace above a sample or air
by SPME is shown in Box 32.14. After sampling, the fibre is retracted within its
holder for protection until inserted in the hot injector of the GC (or mobile phase
of the HPLC); desorption of analytes occurs due to the influence of temperature
in the case of GC or organic solvent in the case of HPLC.
Box 32.13 H
ow to concentrate a sample of an organic compound in an aqueous sample
using direct solid phase microextraction (SPME)
1. Place the sample in a vial. 5. Pierce the GC septum with the SPME needle and then
expose the silica-coated fibre to the hot temperature
2. Clean the SPME fibre by inserting it into the hot injec-
(220°C) of the injection port for 5 min.
tion port (220°C) of the GC.
6. Start the GC isothermal or temperature pro-
3. Expose the SPME fibre to the aqueous sample.
gramme for separation and analysis of the organic
Agitate the sample solution by sonication for 10 min.
compounds.
4. Retract the fibre into the SPME holder.
Box 32.14 H
ow to concentrate a sample of an organic compound in an aqueous sample
using headspace solid phase microextraction (SPME)
1. Place the sample in a septum-sealed vial. 5. Retract the fibre into the SPME holder.
2. Clean the SPME fibre by inserting it into the hot injec- 6. Pierce the GC septum with the SPME needle and
tion port (220°C) of the GC. then expose the fibre to the hot temperature (220°C)
of the injection port for 5 min.
3. Pierce the septum-sealed vial with the needle of the
SPME. 7. Start the GC isothermal or temperature pro-
gramme for separation and analysis of the organic
4. Expose the fibre to the headspace above the aqueous
compounds.
sample for 2 min. Gentle heating of the sample solu-
tion (40°C) may be beneficial.

Chromatography
Key Point Prior to sampling, the silica-coated SPME fibre

should be cleaned. This is done, for example, by exposing the
fibre to the hot injection port of the GC. This is particularly impor-
tant as the fibre can equally adsorb analytes from the atmos-
phere as well as the sample and in some cases the atmosphere
may be the sample.
Purge and trap

Solvent evaporation – Often when the A specialist form of sample preparation is used for volatile compounds in aque-
sample preparation step is complete ous samples, for example, BTEX in aqueous samples. The technique, called
additional steps are required prior to purge and trap, is widely used for the extraction of alcohol in blood samples and
analysis. The most common approach urine followed by gas chromatography. The method involves the introduction of
at this stage is to remove the organic an aqueous sample (e.g. 5 mL urine) into a glass sparging vessel (Fig. 32.45).
solvent. In all cases, the evaporation The sample is then purged with (high purity) nitrogen at a specified flow rate
method is slow to minimise compound and time. The extracted alcohol is then transferred to a trap (e.g. Tenax™)
losses, with a high risk of contamina- at ambient temperature. This is followed by the desorption step. In this step,
tion from the solvent, glassware and the trap is rapidly heated to desorb the trapped alcohol in a narrow band. The
any gas used (see Chapter 18). desorbed alcohol is then transferred via a heated transfer line to the injector of
a gas chromatograph for separation and detection.
Preparation of an air sample
Heaspace analysis – can be done Headspace extraction is a term applied to a range of techniques that sample the
above a solid sample. volume of air above a liquid sample. The nature of the compounds must, by
definition, be volatile and be analysed by gas chromatography.
The concentration of compound(s) in the headspace can be calculated
A volatile compound will be in the using the partition coefficient (K)
headspace above the sample.
K = Cs/Cg[32.12]

To gas
chromatograph
Vent
Multi-port valve
Sample Tenax
loading port trap
Sample
Heater
coil
Purge valve
Purge gas
Carrier gas
(nitrogen)
Injection gas
Fig. 32.45 Purge and trap system for the extraction of volatile compounds
from aqueous samples.

Chromatography
Volatile compound
where Cs is the concentration of the compound in the sample phase (solid
or liquid) and Cg is the concentration of the compound in the gaseous phase
(Figure 32.46). Typical K values for some common solvents are shown in
Gas phase Table 32.3 The value of K is, however, greatly influenced by both temperature
and the addition of salt (so-called salting out). An increase in temperature, for
example raising the temperature of the liquid sample from 40°C to 80°C, will
lower the value of K and hence increase the GC signal response. Similarly the
Sample phase addition of a salt such as, ammonium chloride, ammonium sulphate, sodium
chloride, sodium citrate, sodium sulphate or potassium carbonate, will lower
the value of K and hence increase the GC signal response.
Fig. 32.46 Volatile compound distribution
in a sealed container.
Key Point The addition of salt to the aqueous soution
decreases the solubility of the compounds; as a result the con-
Table 32.3 Typical K values for some com- centration of organic compounds in the headspace incrases.
mon solvents.
Solvent K value
As well as K values, the volume of the sample and gaseous phases is also
Cyclohexane 0.077 important. These are determined using the phase ratio (b).
n-Hexane 0.14
Toluene 2.82

b = Vg/Vs[32.13]
Dichloromethane 5.65
where Vg is the volume of the gaseous phase and Vs is the volume of the
Ethyl acetate 62.4 sample phase. As a result a large sample volume will result in a low b values
Isopropanol 825 producing a higher GC signal response (and vice versa). The ‘salting out’
effect is most significant with compounds that have high K values. Ulti-
mately, seeking to minimise both K and b values will lead to the highest GC
signal response.
Compounds with a low K value will
As well as seeking to optimise K and b values, another way to increase
position more readily into the gaseous
the GC signal response for specific compounds (e.g. alcohols and acids) is by
phase.
the addition of a derivatisation stage. Derivatising the specific compounds will
increase their volatility and hence increase their presence in the headspace
above the sample By decreasing the b value (by increasing sample volume)
Derivatisation – used to modify the compounds with high K values partition less into the headspace compared to
functionality of ar organic compound compounds with low K values; in this situation, it is important to optimise the
so that it can be analysed by G.C. temperature and ‘salting out’ effect first before adjusting the volumes.
Two most common derivatisating Silylation involves the addition of either a trimethyl-silyl group or
reagents are: t- butyldimethylsilyl group to the organic compound using a specific silyating
●● silylation reagent e.g. N, O- bistrimethylsilyl-acetamide (BSA), N,O-bis-trimethylsilyl-
●● acylation trifluoroacetamide (BSTFA), N-methyl-N-trimethylsilyl-trifluoroacetamide
(MSTFA) and N-trimethylsilyimida-zole (TMSI)].
In acylation, the addition of acyl derivatives or acid anhydrides is done
using a range of acylating reagents e.g. tri-fluoroacetic acid (TFAA), pen-
tafluoropropionic acid anhydride (PFPA) and heptafluorobutyric acid anhy-
dride (HFBA)).
Increased gaseous compound presence can also lead to GC issues; specif-
ically poor peak shape, sample carry-over and peak fronting. These issues can
be addressed by the use of cryogenic cooling and sample re-focusing at the
(head) top of the column.
Headspace sampling can be done in a variety of ways, including via a
gas-tight syringe (also referred to as static headspace) or dynamic headspace.
Static headspace analysis (Fig. 32.47) operates by placing the sample vial
in a thermostated oven for a period of time (e.g. an incubation temperature of

Chromatography
60°C for 5 min). Then, a known volume (e.g. 250 mL) of the gaseous head-
space is removed using a gas-tight syringe and injected into the injection port
Gas-tight syringe
of the GC.
Main issues in Static Headspace (SHS) analysis

The main issues to address in the use of SHS are:
●● Procedure is best done using the autosampler of the GC system, fitted with
an incubator.
●● Optimising the b value (i.e. sample to headspace to vial volumes).
●● Whether salting out is required (to improve GC sensitivity).
●● Incubation temperature.
●● Volume of the gas-tight syringe.
Fig. 32.47 Static heaspace analysis. to
allow the opportunity for equilibrium to ●● Potential risk of sample carry-over if a heated gas-tight syringe is not used.
be achieved in the shortest possible time
between the liquid and gaseous phases,
●● Gas-tight syringe is heated to the same temperature as the incubation tem-
shaking of the sample vial is preferred. perature (for the sample vial); prevents sample condensation.
●● Influence of sample injection volume on resultant chromatography.
●● Check the GC injection port septum regularly; the gas-tight syringe has a
Derivatisation – can be done in situ in wider needle than a typical syringe (for GC injection). The potential for a
the sample vial. gas leak to occur is enhanced if the septum is not replaced more regularly.
Static headspace analysis can also be done using solid phase microextrac-
tion, Fig. 32.44(b) and Box 32.14.
Dynamic headspace analysis (Fig. 32.48) operates by placing the sample
vial in a thermostated oven for a period of time (e.g. an incubation temperature
Purge gas
of 80°C) and passing an inert (purge) gas over the surface; the headspace vol-
To GC atile compounds are removed, at a typical purge flow rate of 15 mL/min, and
Trap passed through a trap (e.g. Tenax TA, held at 25°C). The compounds are then
rapidly desorbed from the trap (by heating 25°C to 280°C in 1 min, followed
by a hold of 5 min) directly into the injection port of the GC.
Main issues in Dynamic Headspace (DHS) analysis

The main issues to address in the use of DHS are:
Fig. 32.48 Dynamic headspace analysis. ●● Procedure is best done using a dedicated DHS system.
●● Optimising the b value (i.e. sample to headspace to vial volumes).
●● Whether salting out is required (to improve GC sensitivity).
●● Incubation temperature.
●● Selectivity of the trap (for retaining the compounds).
●● Heated transfer line required; potential for condensation of compounds if
not heated sufficiently.
●● Influence of sample injection volume on resultant chromatography.

Chromatography
Text references
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trometry: An Introduction. John Wiley & Sons Ltd, Sons Ltd, Chichester.
Chichester. Kromidas, S. (2000) Practical Problem Solving in HPLC.
Barry, E.F. and Grob, R.L. (2007) Columns for Gas Chro- John Wiley & Sons Ltd, Chichester.
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Sons Ltd, Chichester. John Wiley & Sons Ltd, New York.
Bertholf, R. and Winecker, R. (2007) Chromatographic McMaster, M. (2008) GC/MS: A Practical User’s Guide,
Separations in Clinical Chemistry and Toxicology. John 2nd edn. John Wiley & Sons Ltd, Chichester.
McNair, H.M. and Miller, J.M. (2009) Basic Gas
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Methods: A Practical Guide. John Wiley & Sons Ltd, Chichester.
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Meloan, C.E. (1999) Chemical Separations: Principles,
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Scott, R.P.W. (1996) Chromatography Detectors: Design,
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Chromatography, 4th edn. John Wiley & Sons Ltd, New
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Harwood, L.M., Moody, C.J. and Percy, J.M. (2000) Exper- & Sons Ltd, Chichester.
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Subramanian, G. (2006) Chiral Separation Techniques: A
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Practical Approach, 3rd edn. John Wiley & Sons Ltd,
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Chromatography
Study exercises
32.1 Calculate the capacity factor, resolution and (a) The resolution factor for the two peaks.
number of theoretical plates of two components (b) The efficiency of each peak.
from a chromatogram. Quinaldine and nicotine
32.4 Calculate Rf values from a chromatogram. The
have retention times of 5.9 and 6.2 min, respec-
figure represents the separation of three pig-
tively on a 30 cm * 0.25 mm id * 0.25 mm film
ments by thin-layer chromatography.
thickness DB5 column. If the peak width of qui-
naldine is 0.16 min and for nicotine is 0.18 min, Origin A B C Solvent front
calculate
(a) Capacity factor for quinaldine and nicotine

(b) Column resolution Thin-layer chromatographic separation of a mix-
(c) Average number of theoretical plates in the ture of three pigments (A, B and C).
column (column efficiency, N) per compound.
What is the RF value of each pigment?
Note: the retention time of the unretained com-
ponent is 1.0 min. Express your answers to three significant figures.
32.2 Test your knowledge of chromatographic 32.5 Test your knowledge of detector terminol-
theory – define the following terms: ogy. Explain what the following acronyms stand
for:
(a) Dead time (to)
(b) Retention time (tR) (a) FID
(c) Capacity factor (k) (b) TCD
(d) Separation factor (a) (c) ECD
(e) Column efficiency (N). (d) DAD.
32.3 Calculate the resolution and efficiency of two 32.6 Check your knowledge of liquid chromatogra-
compounds. Compounds X and Y have retention phy detectors. Make a list of the various major
times of 18.40 and 20.63 min, respectively, on a 30 types of liquid chromatography detector in order,
cm column. If the peak widths at the peak bases from highest to lowest sensitivity. Which of these
are 1.11 and 1.21 min, respectively, calculate: methods is most versatile and why?

33 Electrophoresis
Understanding electrophoresis – this Key Point Electrophoresis is a separation technique based

is, in essence, an incomplete form of on the movement of charged molecules in an electric field. Dis-
electrolysis, since the applied electrical similar molecules move at different rates and the components
field is switched off well before sample of a mixture will be separated when an electric field is applied.
molecules reach the electrodes. It is a widely used technique, particularly for the analysis of
complex mixtures or for the verification of purity (homogeneity)
of isolated molecules.
Definition
While electrophoresis is mostly used for the separation of charged macro-
Electrical field strength – a measure of molecules, techniques are available for high resolution separations of small
the magnitude of an electric field, usually molecules such as amino acids, anions and catecholamines (e.g. by capillary
expressed in terms of electrical potential electrophoresis, p. 299).
difference per unit length, as Vm -1. The electrophoretic mobility of a charged molecule depends on:
Electrophoretic mobility – the rate of ●● Net charge – negatively charged molecules (anions) migrate towards the
migration of a particular type of mole- anode (+), while positively charged molecules (cations) migrate towards the
cule in response to an applied electrical cathode (-); highly charged molecules move faster towards the electrode of
field. opposite charge than those with lesser charge.
●● Size – frictional resistance exerted on molecules moving in a solution means
that smaller molecules migrate faster than large molecules.
●● Shape – the effect of friction also means that the shape of the molecule
will affect mobility, for example globular proteins compared with fibrous
proteins.
●● Electrical field strength – mobility increases with increasing field strength
(voltage), but there are practical limitations to using high voltages, especially
due to heating effects.
The combined influence of net charge and size means that mobility (m) is
Table 33.1 pKa values of ionisable groups determined by the charge : density or the charge : mass ratio, according to the
in selected amino acid residues of proteins. formula:
qE
Group/residue pKa m = [33.1]
r
Terminal carboxyl 3.1
where q is the net charge on the molecule, r is the molecular radius and E is
Aspartic acid 4.4
the field strength.
Glutamic acid 4.4
Histidine 6.5 Electrophoresis and the separation of proteins
Terminal amino 8.0
The net charge of a sample molecule determines its direction of movement and
Cysteine 8.5 significantly affects its mobility. The net charge of a protein molecule is pH
Tyrosine 10.0 dependent, and is determined by the relative numbers of positively and negatively
Lysine 10.0 charged amino acid side chains at a given pH Table 33.1. The degree of ionisation
of each group is pH dependent, resulting in a variation of net charge on the mol-
Arginine 12.0
ecule at different pH values (Fig. 33.1). Thus, electrophoresis is always carried
*Note that these are typical values: the pKa out at constant pH and a suitable buffer must be present along with the sample in
will change with temperature and ionic
order to maintain that pH. For example, if the proteins shown in Fig. 33.1 were
strength. Acidic residues will tend to be neg-
atively charged at pH values above their pKa, subjected to electrophoresis at pH 9.0, and if the proteins were of similar size
while basis residues will tend to be positively and shape, then the rate at which protein A (net charge, -3) migrates towards
charged below their pKa. the anode would be faster than that for protein B (net charge, -1). Separation

Electrophoresis
+3 of proteins is usually carried out at alkaline pH, where most proteins carry a net
negative charge.
+2
Ionisable groups in nucleic acids include the phosphate of the phosphodi-
+1 ester bond and the purine and pyrimidine bases. Most nucleic acid electro-
phoresis is carried out at alkaline pH values at which the negatively charged
net charge
0
2 3 4 5 6 7 8 9 10 phosphate group predominates. Each nucleotide moiety of nucleic acids con-
pH tributes one negatively charged phosphate group, so the charge : mass ratio for
–1
protein B molecules of different sizes will be constant. Nucleic fragments of different
–2 sizes will migrate toward the anode, but separation can only be achieved by
running the sample in a gel that is able to act as a ‘molecular sieve’.
protein A
–3
–4 Basic apparatus
Fig. 33.1 Titration curves for two proteins, Most types of electrophoresis using a supporting medium (described below)
A and B, containing different proportions of are simple to carry out and the apparatus can be easily constructed, although
acidic and basic amino acid residues. inexpensive equipment is commercially available. High-resolution techniques
such as 2D-electrophoresis and capillary electrophoresis require more sophis-
ticated equipment, both for separation and analysis.
Minimising diffusion – make the sam- Simple electrophoretic separations can be performed either vertically
ple zone as narrow as possible, and fix (Fig. 33.2) or horizontally (Fig. 33.3). The electrodes are normally made of
and/or stain the bands as soon as pos- platinum wire, each in its own buffer compartment. In vertical electrophoresis,
sible after the run. the buffer solution forms the electrical contact between the electrodes and the
supporting medium in which the sample separation takes place. In horizontal
electrophoresis electrical contact can be made by buffer-soaked paper ‘wicks’
dipping in the buffer reservoir and laid upon the supporting medium. The buffer
Definition
reservoir normally contains a divider acting as a barrier to diffusion (but not to
Ohm’s law V = IR, where V = voltage,
electrical current), so that localised pH changes which occur in the region of
I = current and R = resistance.
the electrodes (as a result of electrolysis) are not transmitted to the supporting
medium or the sample. Individual samples are spotted onto a solid support-
ing medium containing buffer or are applied to ‘wells’ formed in the supporting
medium. The power pack used for most types of electrophoresis should be
Optimising electrophoresis – attempt- capable of delivering ≈500 V and ≈100 mA.
ing to minimise heat production using
very low currents is not practical, since
Using a supporting medium
it leads to long separation times and
therefore to increased diffusion. The effects of convection currents (resulting from the heating effect of the
applied field) and the diffusion of molecules within the buffer solution can be
minimised by carrying out the electrophoresis in a porous supporting medium.
upper
samples added to wells
This contains buffer electrolytes and the sample is added in a discrete loca-
chamber
tion or zone. When the electrical field is applied, individual sample molecules
buffer
– cathode remain in sharp zones as they migrate at different rates. After separation,
gel cast
between
post-electrophoretic diffusion of selected molecules (e.g. proteins) can be
separated glass plates. avoided by ‘fixing’ them in position on the supporting medium, for example
components Wells are
formed in using trichloracetic acid (TCA).
the top
marker + anode The heat generated during electrophoresis is proportional to the square of
dye the applied current and to the electrical resistance of the medium: even when
buffer a supporting medium is used, heat production will lead to zone broadening by
lower
chamber
increasing the rate of diffusion of sample components and buffer ions. Heat
denaturation of sample proteins may also occur. Another problem is that heat
will reduce buffer viscosity, leading to a decrease in resistance. If the elec-
Fig. 33.2 Apparatus for vertical slab electro- trophoresis is run at constant voltage, Ohm’s law dictates that as resistance
phoresis (components move downwards falls, the current will increase, leading to further heat production. This can
from wells, through the gel matrix). be avoided by using a power pack that provides constant power. In practice,

Electrophoresis
most electrophoresis equipment incorporates a cooling device; even so,

power pack
distortions of an electrophoretic zone from the ideal ‘sharp, linear band’
tank with lid
can often be explained by inefficient heat dissipation.
direction of agarose gel,
submerged in
anode + analyte migration
buffer solution Types of supporting medium
–
cathode These can be subdivided into:
●● Inert media – these provide physical support and minimise convection:
separation is based on charge density only (e.g. cellulose acetate).
●● Porous media – these introduce molecular sieving as an additional
buffer sample effect: their pore size is of the same order as the size of molecules
wells
reservoirs being separated, restricting the movement of larger molecules relative
to smaller ones. Thus, separation depends on both the charge density
Fig. 33.3 Agarose gel electrophoresis.
and the size of the molecule.
With some supporting media (e.g. cellulose acetate), a phenomenon called
electro-endosmosis or electro-osmotic flow (EOF) occurs. This is due to the
Definition
presence of negatively charged groups on the surface of the supporting medium,
Electro-osmotic flow – the osmotically attracting cations in the electrophoresis buffer solution and creating an elec-
driven mass flow of water resulting from trical double layer. The cations are hydrated (surrounded by water molecules)
the movement of ions in an electropho- and when the electric field is applied, they are attracted towards the cathode,
retic system. creating a flow of solvent that opposes the direction of migration of anionic
biomolecules towards the anode. The EOF can be so great that weak anionic
molecules (e.g. antibodies) may be carried towards the cathode.
Handling cellulose acetate – the fragile Where necessary, EOF can be avoided by using supporting media such as
strips must be carefully handled, avoid- agarose or polyacrylamide, but it is not always a hindrance to electrophoretic
ing touching the flat surfaces with your separation. Indeed, the phenomenon of EOF is used in the high resolution
fingers. technique of capillary electrophoresis (p. 299).
Cellulose acetate
CH2OH Acetylation of the hydroxyl groups of cellulose produces a less hydrophilic
CH2
structure than cellulose in the form of paper: as a result it holds less water and
O
OH O diffusion is reduced, with a corresponding increase in resolution. Cellulose
O acetate is often used in the electrophoretic separation of plasma proteins in
O
OH
O clinical diagnosis – it can be carried out quickly (≈45 min) and its resolution
a 1–3 is adequate to detect gross differences in various types of protein (e.g. para-
O glycosidic
OH link proteins in myeloma). Cellulose acetate has a fairly uniform pore structure and
galactose OH
3–6 anhydrogalactose the pores are large enough to allow unrestricted passage of all but the largest of
b 1,4
glycosidic
molecules as they migrate through the medium.
link n
Agarose
Fig. 33.4 Structure of agarose. Additional
sulphate and pyruvyl groups are attached Agarose is the neutral, linear polysaccharide component of agar (from sea-
at selected hydroxyls in the polymer. weed), consisting of repeating galactose and 3,6-anhydrogalactose subunits
(Fig. 33.4). Powdered agarose is mixed with electrophoresis buffer at concen-
trations of 0.5–3.0% w/v, boiled until the mixture becomes clear, poured onto
Advantages of polyacrylamide gels – in a glass plate, then allowed to cool until it forms a gel. Gelation is due to the
addition to their versatility in terms formation of hydrogen bonds both between and within the agarose polymers,
of pore size, these gels are chemically resulting in the formation of pores. The pore size depends on the agarose
inert, stable over a wide range of pH, concentration. Low concentrations produce gels with large pores relative to
ionic strength and temperature, and the size of biological macromolecules, allowing them to migrate relatively
transparent. unhindered through the gel, as determined by their individual charge densi-
ties. Low concentrations of agarose gel are suitable for techniques such as

Electrophoresis
isoelectric focusing (p. 298), where charge is the main basis of separation.
Advantages of polyacrylamide gels – in
The smaller pores produced by higher concentrations of agarose may result
addition to their versatility in terms
in molecular sieving.
of pore size, these gels are chemically
When agarose gels are used for the separation of DNA, the large frag-
inert, stable over a wide range of pH,
ment size means that molecular sieving is observed, even with low concen-
ionic strength and temperature, and
tration gels. This is the basis of the electrophoretic separation of nucleic acids
transparent.
Box 33.1.
Safety Note Preparing gels for Polyacrylamide

PAGE – both acrylamide and bisacryla- Polyacrylamide gel electrophoresis (PAGE) plays a major role in protein
mide are extremely potent neurotoxins, analysis, both for one-dimensional and two-dimensional separations. The gel
so you must wear plastic gloves when is formed by polymerising acrylamide monomer into long chains and cross-
handling solutions containing these linking these chains using N,N′-methylene bisacrylamide (often abbreviated to
reagents. Although the polymerised gel ‘bis’). The process is shown in Fig. 33.5. In most protocols, polymerisation is
is nontoxic, it is still advisable to wear initiated by free radicals produced by ammonium persulphate in the presence
gloves when handling the gel, because of N,N,N′,N′-tetramethylethylenediamine (TEMED). The photodecomposition
some monomer may still be present. of riboflavin can also be used as a source of free radicals.
The formation of polyacrylamide from its acrylamide monomers is
Preparing polyacrylamide gels – most extremely reproducible under standard conditions, and electrophoretic sep-
solutions used for gel preparation can arations are correspondingly precise. The pore size, and hence the extent of
be made in advance, but the ammo- molecular sieving, depends on the total concentration of monomer (% T),
nium persulphate solution must be pre- i.e. acrylamide plus bisacrylamide in a fixed ratio. This means that pores
paied immediately before use. in the gel can be ‘tailored’ to suit the size of biomolecule to be separated:
gels containing 3% acrylamide have large pores and are used in methods
Box 33.1 How to carry out agarose gel electrophoresis of DNA
1. Prepare the gel. Typically, a small volume (10–20 mL) prepare a calibration curve (p. 445), usually by plot-
of buffer plus agarose is heated gently until the pow- ting log10 size (length) against distance travelled.
der dissolves – take care not to overheat, or it will
boil over. Nowadays, gels are often cast with a small 5. Carry out (‘run’) the electrophoresis. DNA separa-
amount of a visualising dye, such as SYBR®Safe. tion is usually carried out at 100–150 V for 30–60 min
(see manufacturer’s instructions for specific details,
2. Prepare the samples. A small amount of sucrose of according to which ‘power pack’ you are using); the
glycerol is usually added, to increase the density of gel should be run until the ‘tracking’ dye has migrated
the sample. A water-soluble anionic ‘tracking’ dye across 80% of the gel.
(e.g. bromophenol blue of xylene cyanol) is also
6. Examine the result. If you have used a visualising
added to each sample, so that migration can be fol-
dye within the agarose, then you simply transfer the
lowed visually.
gel to a UV transiluminator and look for ‘bands’ of
3. Load the samples onto the gel. Individual samples fluorescence corresponding to each DNA fragment.
are added to the performed wells using a pipettor (the
sample should be retained within the well owing to Safety Note – always wear suitable UV-filtering
its higher density, compared with the buffer solution). safety glasses when working with UV radiation to
The volume of sample added to each well is small – protect your eyes. Use a digital camera to photograph
typically less than 25 mL, so a very steady hand and your gel. Alternatively, a dedicated image capture sys-
careful dispensing are needed to pipette each sample tem can be used, e.g. GelDoc®.
accurately.
4. Load the DNA markers onto the gel. Typically, these 7. Extract any DNA bands of interest. If a particular
standards of known size are added to the first and band is required for further study, the piece of gel
last wells of the gel; after electrophoresis, the relative containing that band can be cut from the gel using
positions of the bands of known size can be used to either a clean scalpel or a specialised gel band cutter.

Electrophoresis
O
where molecular sieving should be avoided (e.g. in isoelectric focusing,
p. 298), while higher concentrations of acrylamide (5–30% T) introduce
H2C CH C NH2
molecular sieving to various degrees depending on the size of the sample
Acrylamide components (i.e. with 30% acrylamide gels, molecules as small as Mr 2000
may be subject to molecular sieving). Gels of 6 2.5% are necessary for
free radical mechanism
molecular sieving of molecules of Mr 7 106, but such gels are almost fluid
and require 0.5% agarose to make them solid. Note that a gel of 3% will
CH2 separate DNA by molecular sieving, due to the large size of the nucleic
NH2
CH C acid molecules.
O Before you embark on a particular PAGE protein separation, you will need
CH2
H2N to think about the general strategy for that separation and make certain choices,
C CH including whether to use the following:
O
CH2
NH2 ●● Rod or slab gels – flat slab gels are formed between glass plates, using
CH C
O plastic spacers 0.75–1.5 mm thick: rod gels are made in narrow bore tubes.
CH2 For most separations using several samples, a slab gel saves time because
H2N
C CH up to 25 samples can be separated under identical conditions in a single
O
CH2 gel, while rod gels can only be used for individual samples. Rectangular
NH2 slab gels are also easier to read, by densitometry, and photograph. However,
CH C
O rod gels are useful in preliminary separations, for determining a suitable
Acrylamide polymer
pH and gel concentration, and for applications where the gel is sliced to
extract and assay proteins of interest.
O
●● Dissociating or non-dissociating conditions – the most widely used PAGE
H2C CH C NH protein separation technique uses an ionic detergent, usually sodium dode-
H2C CH C NH cyl sulphate (SDS), which dissociates proteins into their individual pol-
O
ypeptide subunits and gives a uniform net charge along each denatured
N, N’-methylene bisacrylamide polypeptide. This technique, known as SDS-PAGE, requires only micro-
(forms cross-links) gram amounts of sample and is quick and easy to carry out. On the other
hand, if it is necessary to preserve the native protein conformation and bio-
logical activity, non-dissociating conditions are used, i.e. no SDS is added.
In SDS-PAGE the sample protein is normally heated to 100°C for 2 min, in
CH2 buffer containing 1% (w/v) SDS and 1% (w/v) 2-mercaptoethanol, the latter
NH2
HC C to cleave any disulphide bonds. The resultant polypeptides bind to SDS in
O
CH2 CH2 a constant weight ratio, with 1.4 g of SDS per gram of protein. As a result,
H2N NH2 the intrinsic net charge of each polypeptide is ‘swamped’ by the negative
C CH HC C
O O charge imposed by SDS, and there is a uniform negative charge per unit
CH2 H H CH2 length of polypeptide. Since the polypeptides now have identical charge
N N
HC C C CH densities, when they are subject to PAGE (with SDS present) using a gel of
O O appropriate pore size, molecular sieving will occur and they will migrate
CH2 CH2
H2N NH2 strictly according to polypeptide size. This not only gives effective separa-
C CH HC C
O O tion, but the molecular mass of a given polypeptide can be determined by
CH2 comparing its mobility to polypeptides of known molecular mass run under
NH2
HC C the same conditions. Several manufacturers (e.g. Pharmacia, Sigma) supply
O molecular mass standard kits which may include polypeptides of Mr 11 700
cross-linked acrylamide polymer to 212 000 (Table 33.2), together with details of their preparation and use.
Where necessary, the treated sample can be concentrated by ultrafiltration
Fig. 33.5 Reactions involved in the forma-
and the buffer composition can be altered by diafiltration.
tion of polyacrylamide gels.
●● Continuous or discontinuous buffer systems – a continuous system is
Selecting and using gels in 2D electro-
where the same buffer ions are present in the sample, gel and buffer res-
phoresis – a rod gel may be used for
ervoirs, all at the same pH. The sample is loaded directly onto a gel (the
the first dimension and a slab gel for the
‘separating gel’ or ‘resolving gel’) that has pores small enough to introduce
second dimension.
molecular sieving. In contrast, discontinuous systems have different buffers

Electrophoresis
Table 33.2 Molecular masses of standard in the gel compared to the reservoirs, both in terms of buffer ions and pH.
proteins used in electrophoresis. The sample is loaded onto a large-pore ‘stacking gel’, previously polymer-
ised on top of a small-pore separating gel (Fig. 33.6). The individual proteins
Protein Mr log10Mr in the sample concentrate into very narrow zones during their migration
Cytochrome c 11 700 4.068 through the large-pore gel and stack up according to their charge densities,
Myoglobin 17 200 4.236
prior to separation in the small-pore gel, giving enhanced results compared
with continuous systems.
g@globulin (light chain) 23 500 4.371
Carbonic anhydrase 29 000 4.462 ●● pH and buffer for the separation – PAGE can be earned out between pH 3
Ovalbumin 43 000 4.634
and 10. The pH is not critical for continuous SDS-PAGE, since SDS-treated
polypeptides are negatively charged over a wide pH range. However, when
g@globulin (heavy chain) 50 000 4.699
using non-dissociating systems, the pH can be critical, particularly if the
Human albumin 68 000 4.832 biological activity of the molecules is to be retained.
Transferrin 77 000 4.886
●● Gel concentration – at one extreme, a gel with a very high percentage
Myosin (heavy chain) 212 000 5.326
T might totally exclude the sample components, while a gel with a very
low percentage T might lead to all SDS-treated proteins migrating at the
Following the progress of PAGE – add
same rate, i.e. with the electrophoretic front. A sensible approach is to set
bromophenol blue solution (0.002% w/v)
up a series of rod gels in the range 5–15% T and observe the separation and
to the sample in the ratio 1:25 (dye:sam-
resolution obtained. Alternatively, a gradient gel can be used, in which the
ple). This highly ionic, small Mr dye
percentage T increases, and hence pore size decreases, in the direction of
migrates with the electrophoretic front
protein migration. A useful gradient for a preliminary experiment would
be 5–20% T. Such gels are able to resolve protein mixtures with a wide
range of molecular masses. Furthermore, as proteins migrate into regions
Choosing a buffer system for PAGE – of ever-decreasing pore size, the movement of the leading edge of a zone
discontinuous systems are more will become increasingly restricted. This allows the trailing edge to catch
time-consuming to prepare, but have up, resulting in considerable zone sharpening. Gradient formers are fairly
the advantage over continuous systems simple to make, and are commercially available.
in that relatively large volumes of dilute Practical details of the preparation of PAGE and SDS-PAGE gels are given
sample can be used and good resolu- in Table 33.3, while Box 33.2 gives the procedure for SDS-PAGE separation of
tion is still obtained. proteins (see Westermeier, 2004, or Gersten, 1996 for further details).
Choosing a pH for protein electropho-

resis – many proteins have isoelec- Post-electrophoretic procedures – handling of the
tric points in the range pH 4–7 and in supporting medium, staining and analysis
response to electrophoresis with buff-
Handling
ers in the region pH 8.0–9.5, most pro-
teins will migrate towards the anode. All types of supporting medium should be handled carefully: wearing gloves is
advisable, for safety and to avoid transfer of proteins from the skin. Cellulose
acetate strips should be immediately transferred to a fixing and staining solu-
glass cathode
plates – tion. Agarose gels should be dried quickly before staining. Polyacrylamide gels
sample wells in vertical slabs must be freed carefully from one of the glass plates in which
buffer
they are formed, taking care to lever the glass at a point well away from the
anode
stacking part of the gel containing the wells: once free, the gels should be immediately
gel
+ transferred to fixing or staining solution.
separating Rod gels are recovered by a process called ‘rimming’, a technique that
gel
takes a little practice to master. To remove the gel from the tube, hold the tube
buffer in one hand and, using a syringe with a long blunt needle, squirt water between
the gel and the inner wall of the tube: while squirting, move the needle gently
up and down and rotate the tube. If the gel does not become free, perform the
Fig. 33.6 Apparatus for discontinuous

electrophoresis.

Electrophoresis
Table 33.3 Preparation of gels for PAGE and SDS-PAGE. The gel solutions are made by mixing the components in the
proportions and in the order shown. Figures are ml of each solution required to give the stated % gel strength.
PAGE SDS-PAGE
Solution 3.5% gel 5% gel 7.5% gel 5% gel 7.5% gel 10% gel
(added in order shown) (t = 3.6%) (t = 5.1%) (t = 7.7%) (t = 5.1%) (t = 7.7%) (t = 10.2%)
1. Distilled water 19.3 14.9 7.5 14.9 7.5 –

2. TRIS-glycine buffer, pH 33.0 33.0 33.0 – – –
8.9, 0.1 mol L -10.1
3. Imidazole buffer, pH – – – 33.0 33.0 33.0
7.0, 0.1 mol L -1 plus
0.2% w/v SDS
4. Acrylamide solution 10.4 14.8 22.2 14.8 22.2 29.7
22.2% w/v and 0.6%
w/v bis
5. Ammonium persulphate 3.2 3.2 3.2 3.2 3.2 3.2
solution, 0.15% w/v
6. TEMED 0.1 0.1 0.1 0.1 0.1 0.1
Final volume (mL) 66.0 66.0 66.0 66.0 66.0 66.0
same procedure at the other end of the tube, until the gel is released. If neces-
Separating protein mixtures – for high
sary, a Pasteur pipette bulb applied to one end can be used to push the gel out
resolution, a combination of dissociat-
of the tube.
ing and discontinuous PAGE in slab gels
is the system of choice. Fixing, staining and destaining
To prevent separated proteins from diffusing, they are usually fixed in position.
Proteins on cellulose acetate strips can be fixed and stained using Ponceau S
(0.2% w/v in a solution of 3% v/v aqueous trichloracetic acid, TCA).
What to do if your polyacrylamide gels
Fixing is also required for most types of gel electrophoresis: 3% v/v TCA
fail to set – polymerisation is inhib-
is often used. The most widely used stain for protein separations in gels is Coo-
ited by oxygen, so solutions should
massie Blue R-250 (where R = reddish hue): the detection limit is ≈0.2 mg
be degassed, and the surfaces of the
and staining is quantitative up to 20 mg for some proteins. It is normal for
polymerisation mixture exposed to
background staining of the gel to occur, and removal of background colour
air should be overlaid with water; if
(‘destaining’) can be achieved either by diffusion or electrophoresis. To destain
your gels still do not polymerise, the
by diffusion, transfer the gel to isopropanol: acetic acid: water (12.5:10:77.5
most common cause is the use of ‘old’
v/v/v) and allow to stand for 48 h, or change the solution several times to
ammonium persulphate stock solution.
speed up the staining process. Electrophoretic destaining can remove Coo-
If low pH buffers are used, polymerisa-
massie Blue, which is anionic: stained gels are placed between porous plastic
tion may be delayed because TEMED is
sheets with electrodes on each side, and the tank is filled with 7% acetic acid.
required in the free base form.
Passing a current of up to 1.0 A through the gel for around 30–40 min. should
result in substantial destaining.
If you need greater sensitivity (e.g. for nanogram to femtogram amounts),
or when using high resolution techniques such as 2D-electrophoresis (p. 360),
Handling gels – avoid touching gels silver staining can be used. Depending on the protocol chosen and the proteins
with paper as it sticks readily and is dif- being stained, the silver technique can be fivefold to 200-fold more sensitive
ficult to remove without tearing. than Coomassie Blue. The method involves a fixation step (e.g. with TCA),

Electrophoresis
Box 33.2 How to carry out SDS-PAGE for protein separation
1. Prepare the gel. Nowadays, many laboratories use 5. Run the electrophoresis. The gel is positioned with the
pre-cast gels, bought from a manufacturer (e.g. Bio- well/samples closest to th e cathode (negative electrode),
Rad®). If you are pre paring your own gel, you will since they will move towards the anode during electro-
need to follow the protocol very carefully. Typically, phoresis as a result of their negative charge. Protein sep-
the correct proportions of acrvlamide, bisacrylamide aration is typically carried out at 80–100 V for 1–2 h (see
and SDS are mixed together and degassed, under manufacturer’s instructions for specific details, accord-
vacuum.Then ammonium persulphate and tetrameth- ing to which ‘power pack’ you are using): the gel should
ylethylenediamine (TEMED) are added to trigger the be run until the ‘tracking’ dye has migrated across 80%
polymerisation. Once the latter two constituents are of the gel). Higher voltages give faster separation, but
added, the gel should be poured immediately into the poorer separation of protein ‘bands’, and may cause
casting tray (including the well former ‘comb’). denaturation of proteins due to heating.
2. Prepare the samples. The protein sample is mixed 6. Fix and stain the gel. One of the most widely used
with a buffer solution containing SDS (to bind to the approaches uses Coomassie Brilliant Blue stain. Typ-
dissociated proteins), plus dithiothreitol (DTT) (to ically, this involves immersing the gel in 0.25% w/v
cleave disulfide bonds in the proteins) and a ‘tracker’ Coomassie Blue for 1 h then destaining overnight in
dye (e.g. bromophenol blue), then heated for 5 min a methanol/acetic acid solution. A safer, waterbased
95°C, to disrupt the tertiary structure and ‘linearise’ alternative can be bought from a manufacturer (e.g.
the polypeptide chains. Bio-Rad Bio-Safe Coomassie Stain), which allows the
gel to be destained in water overnight. After destain-
3. Load the samples onto the gel. Individual samples ing, separated proteins are visible as blue bands
are added to the wells using a pipettor (p. 64). The against an unstai ned background. For higher resolu-
volume of sample added to each well is typically less tion, a silver stain can be used instead of Coomassie
than 100 mL, so a very steady hand and careful dis- Brilliant Blue.
pensing are needed to pipette each sample accurately
(steady the pipettor using both hands if this helps). 7. Examine the results. After destaining, separated
To optimise the separation of proteins, the volume proteins are visible as blue bands against an
added should be kept as s mall as possible. unstained background. The position of these bands
can be compared with the molecular mass stand-
4. Load the molecular mass standards. Nowadays, ards to determine the size of each band in the test
many labs use ‘rainbow’ markers, with a wide range sample. For greater accuracy, the distance moved by
of proteins of known molecular mass, each of which the molecular mass standards can be plotted against
is sta ined a different colour, to enable estimation of log10 relative molecular mass and the resulting rela-
the molecular masses of unknown proteins, by visual tionship used to determine the size of unknown
comparison. bands.
Avoiding overloaded gels and band followed by exposure to silver nitrate solution and development of the stain.
distortion – determine the protein con- The silver ions are thought to react with basic and thiol groups in proteins, and
centration of the sample beforehand. subsequent reduction (e.g. by formaldehyde at alkaline pH, or by photodevel-
Around 100 mg of a complex mixture, opment) leads to deposits of silver in the protein bands. Most proteins stain
or 1910 mg of an individual component, brown or black, but lipoproteins may stain blue, and some glycoproteins stain
will be sufficient, but bear in mind that yellow or red. Some proteins lacking in amino acids with reducing groups (e.g.
underloading may result in bands being those lacking cysteine residues) may stain negatively, i.e. the bands are more
too faint to be detected. transparent than the background staining of the gel. Although many protocols
have been published, silver stain kits are commercially available, e.g. from
Bio-Rad.
Optimising resolution – keep the sam-
Although silver staining has clear advantages in terms of sensitivity, for
ple volume as small as possible. For ver-
routine work it is more laborious and expensive to carry out than the Coo-
tical slab gels and for rod gels, include
massie Blue method. It also requires high purity water, otherwise signifi-
10% w/v sucrose or glycerol to increase
density and allow buffer solution to be
cant background staining occurs. Another feature is that the staining can be
overlaid on the sample without dilution.
non-specific, since DNA and polysaccharides may stain on the same gel as
proteins.

Electrophoresis
Other methods for the detection of separated components include:

Understanding the terminology of high
purity water – this is usually produced ●● autoradiography for proteins labelled with 32P or 125I;
by reverse osmosis and its purity is
●● fluorescence for proteins pre-labelled with fluorescent dyes;
often expressed with respect to its resis-
tivity: ultrapure water has a resistivity of ●● periodic acid-Schiff (PAS) stain using dansyl hydrazine for glycoproteins.
18 MΩ cm -1 (informally referred to as
‘eighteen megaohm water’). Blotting
The term ‘blotting’ refers to the transfer of separated proteins from the gel
matrix to a thin sheet such as nitrocellulose membrane (commercially available
from, for example, Millipore, Amersham). The proteins bind to this membrane,
and are immobilised. Blotting of proteins is usually achieved by electrophoretic
transfer, and this process is normally referred to as Western blotting. Its major
advantage is that the immobilised proteins on the surface of the membrane
are readily accessible to detection reagents, and stainihg and destaining can
be achieved in less than 5 min. Use of labelled antibodies to detect specific
proteins (immunoblotting) can take less than 6 h. In addition, it is easy to dry
and store Western blots for long periods, for further analysis.
Detection of enzymes
Origins of ‘Western’ blotting – follow- If you need to detect enzyme activity you should use a non-denaturing gel.
ing the description of ‘Southern’ blot- The gel matrix will hinder diffusion of the enzyme, but will allow access to the
ting of DNA other points of the compass small molecular weight substrates, co-factors and dyes necessary to localise
have been used to describe other forms enzyme activity in situ. Most methods for detecting enzymes on gels are mod-
of blotting, with ‘Western’ blotting for ifications of protocols originally developed by histochemists, e.g.:
proteins. ●● NAD+ -requiring oxidoreductases can be detected by incubating the gels
with substrate, NAD+ and a solution of a tetrazolium salt which, when
reduced, forms an insoluble coloured formazan dye.
Example For detection of lactate ●● Transferases and isomerases can be detected by coupling their reactions to
dehydrogenase (LDH): when a solution an oxidoreductase-requiring reaction, visualised as described above.
containing lactate, NAD +, phenazine
methasulfate (PMS) and methyl ●● Hydrolases can be detected using appropriate chromogenic or fluorogenic
thiazolyl tetrazolium (MTT) is added substrates.
to a gel containing LDH, a series of ●● Phosphatases can be detected by precipitating any phosphate released from
redox reactions occurs in the enzyme-
the substrate with Ca2+ .
containing regions, starting with the
oxidation of lactate to p yruvate, and
proceeding via NAD +, PMS and MTT
to the eventual reduction of MTT
Recording and quantification of results
to formazan dye. After incubation A number of expensive, dedicated instruments are available for the analysis of
at 37°C in the dark (since MTT is gels, e.g. laser densitometers. Alternatively, gel scanning attachments can be
light-sensitive), LDH is detected by purchased for standard spectrophotometers, allowing measurement and record-
the appearance of blue–black bands on ing of the absorbance of the Coomassie Blue stained bands at 560–575 nm: for
the gel. instruments connected to a computer, quantification of individual components
can be achieved by integrating the areas under the peaks.
You can photograph gels using a conventional camera (fine grain film),
digital camera, or using a photocopier. Alternatively, a dedicated image capture
and analysis system may be used, e.g. GelDoc. The gel should be placed on
Detecting enzymes in gels – minimise a white glass transilluminator. A red filter will increase contrast with bands
zone spreading by incorporating sub- stained with Coomassie Blue. If the gels themselves need to be retained, they
strates, etc. in a thin agarose indicator can be preserved in 7% acetic acid. Alternatively, they can be dried using a
gel poured over the separating gel. commercially available gel dryer.

Electrophoresis
Although the resolution obtained using the basic electrophoretic tech-

Measuring peak areas – a valid ‘low-
niques is adequate for many biomolecular applications, a number of advanced
tech’ alternative to computer-based
techniques are available that give very high resolution with very small amounts
systems is to cut out and weigh peaks
of sample material.
from a recorder chart.
Isoelectric focusing (IEF)

In contrast to electrophoresis, which is carried out at constant pH, IEF is carried
+3 out using a pH gradient. The gradient is formed using small molecular mass
ampholytes, which are analogues and homologues of polyamino-, polycar-
+2
boxylic acids that collectively have a range of isoelectric points (pI values)
+1 between pH 3 and 10. The mixture of ampholytes (p. 113), either in a gel or in
free solution, is placed between the anode in acid solution (e.g. H3PO4), and
net charge
0
2 3 4 5 6 7 8 9 10 the cathode in alkaline solution (e.g. NaOH). When an electric field is applied,
pH each ampholyte migrates to its own pI and forms a stable pH gradient which
–1
Y will persist for as long as the field is applied. When a protein sample is applied
–2 to this gradient, separation is achieved, since individual proteins will migrate
to their isoelectric points. The net charge on the protein when first applied
X
–3 will depend on the specific ‘titration curve’ for that protein (Fig. 33.7). As an
example, consider two proteins, X and Y, having pI values of pH 5 and pH 8
Fig. 33.7 Titration curves for two proteins, respectively, which are placed together on the gradient at pH 6 (Fig. 33.8). At
X and Y. that pH, protein X will have a net negative charge, and will migrate towards
the anode, progressively losing charge until it reaches its pI (pH 5) and stops
migrating. Protein Y will have a net positive charge at pH 6, and so will migrate
+ anode
towards the cathode until it reaches its pI (pH 8).
Using a polyacrylamide gel as a supporting medium and a narrow pH
3
gradient, proteins differing in pI by 0.01 units can be separated. Even greater
4 protein X migrates resolution is possible in free solution (e.g. in capillary electrophoresis, p. 299).
to pH 5.0
Such resolution is possible because protein molecules that diffuse away from
5
the pI will acquire a net charge (negative at increased pH, positive at decreased
proteins X and Y
6 applied to pH) and immediately be focused back to their pI. This focusing effect will
pH gradient
gradient at pH 6.0 continue for as long as the electric field is applied.

7
A useful variant of IEF is in obtaining titration curves for proteins. A
8 pH gradient is set up, and the sample applied in a line at a right angle to the
protein Y migrates gradient. The net charge on a given protein will vary according to its position
9
to pH 8.0 on the gradient – when electrophoresis is carried out at right angles to the pH
10 gradient, the protein will migrate at a velocity and direction governed by that
charge. When stained, each protein will appear as a continuous curved line,
11
corresponding to its titration curve. This technique can be usefully performed,
– cathode during protein purification, prior to ion-exchange chromatography (p. 251): by
obtaining the titration curve for a protein of interest and those of major con-
Fig. 33.8 The migration of two proteins, X taminants, the mobile phase pH that gives optimal separation can be selected.
and Y, in response to a pH gradient. In IEF, it is important that electro-osmotic flow (EOF, p. 291) is avoided,
as this would affect the ability of the proteins to remain stationary at their pIs.
For gel IEF, polyacrylamide minimises EOF, while capillary IEF uses narrow
Avoiding streaking in 2D electro bore tubing with an internal polymer coating (p. 251).
phoresis – ensure that the sample con-
tains no particulate material (e.g. from
protein aggregation); filter or centrifuge Two-dimensional electrophoresis
before use. The most commonly used version of this high resolution technique involves
separating proteins by charge in one dimension using IEF in polyacrylamide
gel, followed by separation by molecular mass in the second dimension using

Electrophoresis
denaturing SDS-PAGE (p. 293). The technique allows up to 1000 proteins

Maximising resolution in 2D electro-
to be separated from a single sample. Typically, the first dimension IEF run
phoresis of proteins – try to minimise
(pH 3–10) is carried out on gel strips of length 7–24 cm. Strips are run at a
nucleic acid contamination of your
voltage of 500–3500 V for 1.5 h, then at 3500 V for a further 4 h. Gel strips can
sample, as they may interact with poly-
then be used immediately, or frozen until required.
peptides/proteins, affecting their move-
It is common for the second-dimension SDS-PAGE separation to be car-
ment in the gel.
ried out on a discontinuous slab gel 0.5–1.5 mm thick, which includes a low
percentage T stacking gel and a separating gel with an exponential gradient
of 10–16% T. The separating gel can be prepared in advance, but the stack-
Freezing gel strips – be sure to mark ing gel must be formed shortly before addition of the rod gel from the one
the identity and orientation of each gel dimensional run.
strip before freezing, e.g. by inserting a
After equilibration with the buffer used in SDS-PAGE, the IEF gel strip is
fine wire into one end of the strip. Note
loaded onto the 2D gel (still between the glass plates in which it was formed)
that if urea is used in the gel strip, it will
and sealed in position using acrylamide or agarose. Before the sealing gel
form crystals on freezing.
sets, a well should be formed in it at one end to allow addition of molecular
mass markers. The second-dimension is run at 100–200 V until the dye front
is ≈1 cm from the bottom edge of the slab. After running, the gel is processed
low high for the detection of polypeptides, e.g. using Coomassie Blue or silver stain.
pH IEF pH Analysis of the complex patterns that result from 2D electrophoresis requires
computer-aided gel scanners to acquire and process data from a gel image, such
as that shown in Fig. 33.9. These systems can compare, adjust and match up
patterns from several gels, allowing both accurate identification of spots and
quantification of individual proteins. Allowance is made for the slight variations
SDS – PAGE
in patterns found in different runs, using internal references (‘landmarks’),

which are either added standard proteins or particular spots known to be present
in all samples.
Capillary electrophoresis (CE)

This technique combines the high resolving power of electrophoresis with the
speed and versatility of HPLC (p. 265). The technique largely overcomes the
major problem of carrying out electrophoresis without a supporting medium,
i.e. poor resolution caused by convection currents and diffusion. A capillary
tube has a high surface area: volume ratio, and consequently the heat gener-
Fig. 33.9 Two-dimensional separation of ated as a result of the applied electric current is rapidly dissipated. A further
proteins from 100x concentrated urine advantage is that very small sample volumes (5–10 nl) can be analysed. The
(2.5 mg total protein; silver stain). versatility of CE is demonstrated by its use in the separation of a range of bio-
Courtesy of T. Marshall and K.M. Williams. molecules, e.g. amino acids, proteins, nucleic acids, drugs, vitamins, organic
acids and inorganic ions; CE can even separate neutral species, e.g. steroids,
aromatic hydrocarbons (see Weinberger, 2000).
The components of a typical CE apparatus are shown in Fig. 33.10. The
capillary is made of fused silica and externally coated with a polymer for
mechanical strength. The internal diameter is usually 25950 mm, a compromise
between efficient heat dissipation and the need for a light path that is not too
short for detection using UV/visible spectrophotometry. A gap in the polymer
coating provides a window for detection purposes. Samples are injected into the
capillary by a variety of means, e.g. electrophoretic loading or displacement.
In the former, the inlet end of the capillary is immersed in the sample and a
pulse of high voltage is applied. The displacement method involves forcing
the sample into the capillary, either by applying pressure in the sample vial
using an inert gas, or by introducing a vacuum at the outlet. The detectors used
in CE are similar to those used in chromatography (p. 267), e.g. UV/visible

Electrophoresis
high voltage spectrophotometric systems. Fluorescence detection is more sensitive, but this
supply
may require sample derivatisation. Electrochemical and conductivity detection
is also used in some applications, e.g. conductivity detection of inorganic cat-
ions such as Na+ and K+ .
Electro-osmotic flow (EOF), described on page 291, is essential to the
most commonly used types of CE. The existence of EOF in the capillary is the
capillary result of the net negative charge on the fused silica surface at pH values over
coil 3.0. The resulting solvent flow towards the cathode is greater than the attraction
of anions towards the anode, so they will flow towards the cathode (note that
detector
the detector is situated at the cathodic end of the capillary). The greater the net
–
+ negative charge on an anion, the greater is its resistance to the EOF and the
lower its mobility. Separated components migrate towards the cathode in the
outlet vial order: (1) cations, (2) neutral species and (3) anions.
inlet vial
Capillary zone electrophoresis (CZE)
data acquisition
This is the most widely used form of CE, and is based on electrophoresis in free
and analysis solution and EOF, as discussed above. Separations are caused by the charge:
mass ratio of the sample components, and the technique can be used for almost
Fig. 33.10 Components of a capillary elec- any type of charged molecule, and is especially useful for peptide separation
trophoresis system. and confirmation of purity.
Micellar electrokinetic chromatography (MEKC)

This technique involves the principles of both electrophoresis and chromatog-
raphy. Its main strength is that it can be used for the separation of neutral mol-
ecules as well as charged ones. This is achieved by including surfactants (e.g.
SDS, Triton X-100) in the electrophoresis buffer at concentrations that promote
the formation of spherical micelles, with a hydrophobic interior and a charged,
hydrophilic surface. When an electric field is applied, these micelles will tend
to migrate with or against the EOF, depending on their surface charge. Anionic
surfactants like SDS are attracted by the anode, but if the pH of the buffer is
high enough to ensure that the EOF is faster than the migration velocity of the
Distinguishing between stereoi- micelles, the net migration is in the direction of the EOF, i.e. towards the cath-
somers – the R and S convention ode. During this migration, sample components partition between the buffer
involves prioritising atoms or groups and the micelles (acting as a pseudo-stationary phase); this may involve both
bonded to a chiral carbon atom (one hydrophobic and electrostatic interactions. For neutral species it is only the
which has four different atoms or partitioning effect that is involved in separation; the more hydrophobic a sam-
groups attached) in order of their ple molecule, the more it will interact with the micelle, and the longer will be
atomic number. With the smallest atom its migration time, since the micelle resists the EOF. The versatility of MEKC
or group pointing away from you, note enables it to be used for separations of biomolecules as diverse as amino acids
the size of the three remaining atoms or and polycyclic hydrocarbons. MEKC is also known as micellar electrokinetic
groups. If the configuration of increas- capillary chromatography (MECC).
ing atomic number is clockwise, this is
termed the (R)-configuration (L. rectus); Chiral capillary electrophoresis (CCE)
if the order is anticlockwise, this is the Resolution of a pair of chiral enantiomers (optical isomers) represents one of
(S)-configuration (L. sinister). Note that the biggest challenges for separation science, because each member of the pair
the older, alternative terminology of will have identical physicochemical properties. CE offers an effective method
(D) and (L) stereoisomers is still widely of separating enantiomers by inducing a ‘chiral selector’ in the electrophoresis
used but the two systems do not always medium. The most commonly used chiral selectors are cyclodextrins such as
coincide [i.e. (R) is not always (D) and the highly sulfated cyclodextrins (HSCDs) (Fig. 33.11). As the enantiomers
(S) is not always L]. migrate along the capillary, one will tend to interact more strongly than the

Electrophoresis
OH OSO3–
OSO3– OH
OH
OSO3– 1.0
R
OSO3– OH
OH OSO3– S CH3
OSO3– OH
NH2
amphetamine
O O
O internal
0.5
–O SOCH
3 2
CH2OSO3– standard
A200
CH2OSO3–
Fig. 33.11 General structure of highly sul-

fated cyclodextrins (HSCDs). The central
cavity of the cyclodextrin (shaded blue)
can interact differently with the R and 5
isomers of a biomolecule, enabling sepa-
ration in CE.
Copyright © 1909–2002 Beckman Coulter, Inc.
http://www.sciex.com/ion-analysis/chiral-analysis
0
0 5 10
Time (min)
Fig. 33.12 CE separation of R and S enantiomers of amphetamine using

Beckman Coulter SCIEX highly sulfated gamma cyclodextrin.
Copyright © 1909–2002 Beckman Coulter, Inc. http://www.sciex.com/ion-analysis/
chiral-analysis
other and its mobility will be reduced relative to the other. Figure 33.12 shows
separation of the R and S forms of amphetamine using 5% HSCD in the elec-
trophoresis buffer. The R-form has greater affinity for the HSCD used, so its
retention time on the capillary is longer than that of the S-form.
Note that HSDCs can also be used in HPLC, but CCE is more effective,
with shorter development times and lower reagent costs.
[Note: alternatively, d and e are replaced by (+) or (-), respectively.]
Capillary gel electrophoresis (CGE)

The underlying principle of this technique is directly comparable with that
of conventional PAGE, i.e. the capillary contains a polymer that acts as a
molecular sieve. As charged sample molecules migrate through the polymer
network, larger molecules are hindered to a greater extent than smaller ones
and will tend to move more slowly. CGE differs from CZE and MEKC in
that the inner surface of the capillary is polymer-coated to prevent EOF; this
means that for most applications (e.g. polypeptide or oligonucleotide separa-
tions) sample components will migrate towards the anode at a rate detennined
by their size. The technique also differs from conventional PAGE in that a

Electrophoresis
‘polymer network’ is used rather than a gel: the polymer network may be
Choosing a detector for capillary elec-
polyacrylamide or agarose.
trophoresis – most types of HPLC
CGE offers the following advantages over conventional electrophoresis:
detector are suitable for CE and related
applications (see Chapter 32). ●● efficient heat dissipation means that a high electrical field can be applied,
giving shorter separation times;
●● detection of the separated components as they move towards the anodic
end of the capillary (e.g. using a UV/visible detector) means that staining
is unnecessary;
●● automation is feasible.
Capillary isoelectric focusing (CIEF)

This is used mainly for protein separation. Here, the principles of IEF are
valid as long as EOF is prevented by using capillaries that are polymer-coated
on their inner surface. Sample components migrate to their isoelectric points
and become stationary. Once separated (6 10 min), the components must
be mobilised so that they flow past the detector. This is achieved by chang-
ing the NaOH solution in the cathodic reservoir with an NaOH/NaCl solu-
tion. When the electric field is reapplied, Cl- enters the capillary, causing a
decrease in pH at the cathodic end and the subsequent migration of sample
components.
Text references
Gersten, D. (1996) Gel Electrophoresis: Proteins (Essential Weinberger, R. (2000) Practical Capillary Electrophoresis.
Techniques Series). John Wiley & Sons Ltd, New York. Academic Press, New York.
Manchenko, G.P. (2002) Handbook of Detection of Westermeier, R. (2004) Electrophoresis in Practice A
Enzymes on Electrophoretic Gels, 2nd edn. CRC Press, Guide to Methods and Applications of DNA and Protein
Boca Raton. Separations, 4th edn. Wiley VCH, Berlin.

Anon. The American Electrophoresis Society Homepage. Schmitt-Kopplin, P. (2010) Capillary Electrophoresis:
Available: http://www.aesociety.org Methods and Protocols. Humana, New York.
Streage, M.A. and Lagu, A.L. (2004) Capillary Electro-
Anon. Cyclodextrin Resource. Available: http://www.cyclo- phoresis of Proteins and Peptides. Humana Press, New
dex.com/index.html Jersey.
Rabilloud, T. (2004) Proteome Research Two-dimensional
Anon. British Society for Proteomics Research Homepage. Gel Electrophoresis and Identification Methods.
Available: http://www.bspr.org/ Springer, Heidelberg.
[Formerly the British Electrophoresis Society, includes Tietz, D. (1998) Nucleic Acid Electrophoresis Springer-
links to other websites and databases.] Verlag. Berlin.
Rabilloud, T. (2004) Proteome Research Two-dimensional Westermeier, R. (2016) Electrophoresis in Practice: A
Gel Electrophoresis and Identification Methods. Guide to Methods and Applications of DNA and Protein
Springer, Heidelberg. Separations, 5th edn. Blackwell Verlog, Berlin.

Electrophoresis
Study exercises
33.1 Find out why the net charge on a protein mol- 33.2 Consider the requirements for sample applica-
ecule varies with pH. Identify the amino acids tion in PAGE and IEF. Explain why in PAGE the
primarily responsible for determining the net sample is applied in a discrete narrow band, usu-
charge on a protein molecule and draw simple ally at the cathodic end of the gel, while in IEF
diagrams to represent the ionisation of their side the sample can be applied at any point along the
chains, indicating how you would expect these length of the gel without concern about location
side chains to be charged at acid, neutral and of narrowness of the sample zone.
alkaline pH values.

34 Electroanalytical techniques
Electrochemical methods are used to quantify a broad range of different mol-

Definitions ecules, including ions, gases, metabolites and drugs.
Oxidation – loss of electrons by an
atom or molecule (or gain of O atoms, Key Point The basis of all electrochemical analysis is the
loss of H atoms, increase in positive transfer of electrons from one atom or molecule to another atom
charge, or decrease in negative charge). or molecule in an obligately coupled oxidation--reduction reac-
Reduction – gain of electrons by an tion (a redox reaction).
atom or molecule (or loss of O atoms,
gain of H atoms, decrease in positive
charge or increase in negative charge).
It is convenient to separate such redox reactions into two half-reactions and, by
convention, each is written as:
reduction
oxidised form + electron (s)(ne- ) oxidation
∆ reduced form[34.1]
You should note that the half-reaction is reversible: by applying suitable con-
ditions, reduction or oxidation can take place. As an example, a simple redox
voltmeter
reaction occurs when metallic zinc (Zn) is placed in a solution containing cop-
per ions (Cu2+ ), as follows:
direction of
electron
flow Cu2+ + Zn S Cu + Zn2+ [34.2]
The half-reactions are (i) Cu2+ + 2e - S Cu and (ii) Zn2+ + 2e - S Zn. The
oxidising power of (i) is greater than that of (ii), so in a coupled system, the lat-
ter half-reaction proceeds in the opposite direction to that shown above, i.e. as
Zn - 2e - S Zn2+ . When Zn and Cu electrodes are placed in separate solutions
Zn2+ Cu2+
containing their ions, and connected electrically (Fig. 34.1), electrons will flow
SO42– SO42– from the Zn electrode to Cu2+ via the Cu electrode owing to the difference in
oxidising power of the two half-reactions.
By convention, the electrode potential of any half-reaction is expressed rel-
zinc + KCl
–
copper ative to that of a standard hydrogen electrode (half-reaction 2H + + 2e - S H2)
anode salt bridge cathode
and is called the standard electrode potential, E. Table 34.1 shows the values
Fig. 34.1 A simple galvanic electro-chemical of E 0 for selected half-reactions. With any pair of half-reactions from this
cell. The KCl salt bridge allows migration of series, electrons will flow from that having the lowest electrode potential to
ions between the two compartments but that of the highest. E 0 is determined at pH = 0. It is often more appropriate
prevents mixing of the two solutions. to express standard electrode potentials at pH7 for biological systems, and
the symbol E 0′ is used: in all circumstances, it is important that the pH is
clearly stated.
The arrangement shown in Fig. 34.1 represents a simple galvanic cell
where two electrodes serve as the interfaces between a chemical system and
an electrical system. For analytical purposes, the magnitude of the potential
(voltage) or the current produced by an electrochemical cell is related to the
concentration (strictly the activity, a, p. 104) of a particular chemical species.
Electrochemical methods offer the following advantages:
Definition
●● excellent detection limits, and wide operating range (10-1 to 10-8 mol L-1);
Galvanic cell – an electrochemical cell
●● measurements may be made on very small volumes (mL) allowing small
in which reactions occur spontaneously
amounts (pmol) of sample to be measured in some cases;
at the electrodes when they are con-
nected externally by a conductor, pro- ●● miniature electrochemical sensors can be used for certain in vivo measure-
ducing electrical energy. ments, e.g. pH, glucose and oxygen content.

Electroanalytical techniques
Table 34.1 Standard electrode potentials* Potentiometry and ion-selective electrodes

(E0) for selected half-reactions
Operating principles
Half-reaction 0
E at 25°C These systems involve galvanic cells (p. 304) and are based on measurement
(V) of the potential (voltage) difference between two electrodes in solution when
Cl2 + 2e - ∆ 2 Cl - + 1.36 no net current flows between them: no net electrochemical reaction occurs and
measurements are made under equilibrium conditions. These systems include
+
O2 + 4H + 4e ∆ 2H2O - + 1.23
methods for measuring pH, ions, and gases such as CO2 and NH3. A typical
- -
Br2 + 2e ∆ 2Br + 1.09 potentiometric cell is shown in Fig. 34.2. It contains two electrodes:
+
Ag + e ∆ Ag - + 0.80
1. a ‘sensing’ electrode, the half-cell potential of which responds to changes
Fe 3+ -
+ e ∆ Fe 2+ + 0.77 in the activity (concentration) of the substance to be measured; the most
I3- + 2e - ∆ 3I - + 0.54 common type of indicator electrodes are ion-selective electrodes (ISEs);
Cu 2+
+ 2e ∆ Cu - + 0.34 2. a ‘reference’ electrode, the potential of which does not change, forming
Hg2Cl2 + 2e ∆ 2Hg + 2Cl - -
+ 0.27
the second half of the cell.
-
AgCl + e ∆ Ag + Cl -
+ 0.22 To assay a particular analyte, the potential difference between these electrodes
+ 0.01
is measured by an mV meter (e.g. a standard pH meter).
Ag(S2O3)3- -
2 + e ∆ Ag +
2S2O2- Reference electrodes for potentiometry are of three main types:
3
2H+ + 2e - ∆ H2 + 0.00 1. The standard hydrogen electrode, which is the reference half-cell
-
AgI + e ∆ Ag + I - 0.15
electrode, defined as 0.0 V at all temperatures, against which values of E 0
are expressed. H2 gas at 1 atmosphere pressure is bubbled over a p latinum
PbSO4 + 2e ∆ Pb + -
SO2- - 0.35
4 electrode immersed in an acid solution with an a ctivity of unity. This elec-
Cd2+ + 2e - ∆ Cd - 0.40 trode is rarely used for analytical work, since it is unstable and other ref-
- 0.76 erence electrodes are easier to construct and use.
Zn2+ + 2e - ∆ Zn
2. The calomel electrode (Fig. 34.3), which consists of a paste of mercury
*From Milazzo et al. (1978).
covered by a coat of calomel (Hg2Cl2), immersed in a saturated solution
of KCl. The half-reaction Hg2Cl2 + 2e - S 2 Hg + 2Cl- gives a stable
standard electrode potential of +0.24 V.
Using a calomel electrode – always
3. The silver/silver chloride electrode. This is a silver wire coated with
ensure that the KCl solution is satu-
AgCl and immersed in a solution of constant chloride concentration. The
rated by checking that KCl crystals are
half-reaction AgCl + e - S Ag + Cl- gives a stable, standard electrode
present.
potential of +0.20 V.
pH/mV meter
Key Point Ion-selective electrodes (ISEs) are based on meas-
urement of a potential across a membrane which is selective for
a particular analyte.
An ISE consists of a membrane, an internal reference electrode, and an internal

reference electrolyte of fixed activity. The ISE is immersed in a sample solution
test solution that contains the analyte of interest, along with a reference electrode. The mem-
brane is chosen to have a specific affinity for a particular ion, and if activity of
sensing electrode
this ion in the sample differs from that in the reference electrolyte, a potential
reference
electrode stirrer bar develops across the membrane that is dependent on the ratio of these activities.
Since the potentials of the two reference electrodes (internal and external) are
magnetic fixed, and the internal electrolyte is of constant activity, the measured potential,
stirrer
E, is dependent on the membrane potential and is given by the Nernst equation:
RT
Fig. 34.2 Components of a potentiometric E = K + 2.303 log [a][34.3]
cell. zF

where K represents a constant potential which is dependent on the reference

Using ISEs, including pH electrodes –
electrode, z represents the net charge on the analyte, [a] the activity of analyte
standards and samples must be meas-
in the sample and all other symbols and constants have their usual meaning
ured at the same temperature, as
(p. 40). For a series of standards of known activity, a plot of E against log
the Nernst equation shows that the
[a] should be linear over the working range of the electrode, with a slope of
measured potential is temperature
2.303RT/zF (0.059V at 25°C). Although ISEs strictly measure activity, the
dependent.
potential differences can be approximated to concentration as long as (i)the
analyte is in dilute solution (p. 104), (ii) the ionic strength of the calibration
standards matches that of the sample, e.g. by adding appropriate amounts of a
electrical
lead
high ionic strength solution to the standards, and (iii) the effect of binding to
sample macromolecules (e.g. proteins, nucleic acids) is minimal. Potentiom-
etric measurements on undiluted biological fluids, e.g. K+ and Na+ levels in
plasma, tissue fluids or urine, are likely to give lower values than flame emis-
outer tube
sion spectrophotometry, since the latter procedure measures total ion levels,
saturated
rather than just those in aqueous solution.
KCl solution All of the various types of membrane used in ISEs operate by incorporating
the ion to be analysed into the membrane, with the accompanying establish-
inner tube ment of a membrane potential. The scope of electrochemical analysis has been
filler hole and
rubber bung extended to measuring gases and non-ionic compounds by combining ISEs
Hg/Hg2Cl2 paste
with gas-permeable membranes, enzymes, and even immobilised bacteria or
tissues.
Glass membrane electrodes
small hole in The most widely used ISE is the glass membrane electrode for pH measure-
inner tube
ment (p. 113). The membrane is thin glass (50 mm wall thickness) made of
KCl crystals silica which contains some Na+ . When the membrane is soaked in water, a thin
sintered glass disk hydrated layer is formed on the surface in which negative oxide groups (Si@O- )
in the glass act as ion-exchange sites. If the electrode is placed in an acid solu-
Fig. 34.3 A calomel reference electrode. tion, H + exchanges with Na+ in the hydrated layer, producing an external sur-
face potential: in alkaline solution, H + moves out of the membrane in exchange
for Na+ . Since the inner surface potential is kept constant by exposure to a fixed
activity of H + , a consistent, accurate potentiometric response is observed over
a wide pH range. Glass electrodes for other cations (e.g. Na+ , NH4+ ) have been
developed by changing the composition of the glass, so that it is predominantly
sensitive to the particular analyte, though the specificity of such electrodes is
not absolute. The operating principles and maintenance of such electrodes are
broadly similar to those for pH electrodes (p. 113).
Gas-sensing glass electrodes
Here, an ISE in contact with a thin external layer of aqueous electrolyte (the
‘filling solution’) is kept close to the glass membrane by an additional, outer
membrane that is selectively permeable to the gas of interest. The arrangement
pH-sensitive
pH electrode glass membrane for a CO2 electrode is shown in Fig. 34.4: in this case the outer membrane is
made of CO2@permeable silicone rubber. When CO2 gas in the sample selec-
tively diffuses across the membrane and dissolves in the filling solution (in this
H+
filling solution
case an aqueous NaHCO3/NaCl mixture), a change in pH occurs owing to the
–+
CO2+H2O H2CO3 HCO3 (NaHCO3 + NaCl) shift in the equilibrium:
CO2 + H2O ∆ H2CO3 ∆ H + + HCO3- [34.4]

CO2 sample
CO2 -permeable The pH change is ‘sensed’ by the internal ion-selective pH electrode, and its
membrane
response is proportional to the partial pressure of CO2 of the solution (PCO2).
Fig. 34.4 Underlying principles of a A similar principle operates in the NH3 electrode, where a Teflon® membrane
gas-sensing electrode. is used, and the filling solution is NH4Cl.

Liquid and polymer membrane electrodes

Using CO2 electrodes – applications
include measurement of blood PCO2 In these types of ISEs, the liquid is a water-insoluble viscous solvent containing
and in enzyme studies where CO2 is a soluble ionophore, i.e. an organic ion exchanger, or a neutral carrier mole-
utilised or released: calibration of the cule, that is specific for the analyte of interest. When this liquid is soaked into
electrode is accomplished using 5% v/v a thin membrane such as cellulose acetate, it becomes effectively immobilised.
and 10% v/v mixtures of CO2 in an inert The arrangement of analyte (A+ ) and ionophore in relation to this membrane
gas equilibrated against the measuring is shown in Fig. 34.5. The potential on the inner surface of the membrane is
solution. kept constant by maintaining a constant activity of A+ in the internal solution,
so the potential change measured is that which results from A+ in the sample
interacting with the ionophore in the outer surface of the membrane.
A relevant example of a suitable ionophore is the antibiotic valinomycin,
which specifically binds K+ . Other ionophores have been developed for meas-
urement of, for example, NH4+ , Ca2+ , Cl- . In addition, electrodes have been
developed for organic species by using specific ion-pairing reagents in the
membrane that interact with ionic forms of the organic compound, e.g. with
drugs such as 5,5-diphenylhydantoin.
Solid-state membrane electrodes

Definition
These contain membranes made from single crystals or pressed pellets of salts
Ionophore – a compound that enhances of the analyte. The membrane material must show some permeability to ions
membrane permeability to a specific and must be virtually insoluble in water. Examples include:
ion: an ionophore may be incorporated
into an ISE to detect that ion.
●● the fluoride electrode, which uses LaF3 impregnated with Eu2+ (the latter to
increase permeability to F - ). A membrane potential is set up when F - in the
sample solution enters spaces in the crystal lattice;
●● the chloride electrode, which uses a pressed pellet membrane of Ag2S and AgCl.
Definitions Voltammetric methods

Electrolytic cell – an electrochemical Voltammetric methods are based on measurements made using an electro-
cell in which reactions are driven by chemical cell in which electrolysis is occurring. Voltammetry, sometimes also
the application of an external voltage called amperometry, involves the use of a potential applied between two elec-
greater than the spontaneous potential trodes (the working electrode and the reference electrode) to cause oxidation
of the cell. or reduction of an electroactive analyte. The loss or gain of electrons at an
Electrolysis – a non-spontaneous electrode surface causes current to flow, and the size of the current (usually
chemical reaction resulting from the measured in mA or mA) is directly proportional to the concentration of the
application of a potential to an electrode. electroactive analyte. The materials used for the working electrode must be
internal solution
A+ X–
– – – –
Em (internal)
C + + + +
CA+ CA+ membrane
C – – – –
Em (external)
+ + + +
A+ X–
external (sample) solution
Fig. 34.5 Underlying principles of a liquid membrane ion-selective elec-

trode. A + = analyte; C = neutral carrier ionophore; Em = surface poten-
tial; membrane potential = Em(internal) - Em(external).

good conductors and electrochemically inert, so that they simply transfer elec-
trons to and from species in solution. Suitable materials include Pt, Au, Hg
and glassy carbon.
Two widely used devices that operate on the voltammetric principle are
the oxygen electrode and the glucose electrode. These are sometimes referred
to as amperometric sensors.
Oxygen electrodes
The Clark (Rank) oxygen electrode
These instruments measure oxygen in solution using the polarographic prin-
ciple, i.e. by monitoring the current flowing between two electrodes when a
voltage is applied. The most widespread electrode is the Clark type (Fig. 34.6),
manufactured by Rank Bros, Cambridge, UK, which is suitable for measur-
ing O2 concentrations in cell, organelle and enzyme suspensions. Pt and Au
electrodes are in contact with a solution of electrolyte (normally saturated
KCl). The electrodes are separated from the medium by a Teflon® membrane,
permeable to O2. When a potential is applied across the electrodes, this gen-
erates a current proportional to the O2 concentration. The reactions can be
summed up as:
4Ag S 4Ag + + 4e - (at silver anode)

O2 + 2e - + 2H + ∆ H2O2 (in electrolyte solution; O2 replenished
by diffusion from test solution)
H2O2 + 2e - + 2H + ∆ 2H2O (at platinum cathode)
leads to battery
and recorder stopper (volume
adjuster)
locking nut
rubber ‘O’ ring
incubation
chamber
magnetic
filler (plus plug) stirrer bar
rubber ‘O’
ring
saturated KCl + to
central – recorder
silver
anode platinum
Teflon cathode circular saturated
membrane silver KCl solution
anode
glass electrode body
Fig. 34.6 Transverse section through a Clark (Rank) oxygen electrode.

rubber 'O'
ring Teflon membrane
platinum Oxygen probes
cathode
Clark-type oxygen electrodes are also available in probe form for immer-
Fig. 34.7 A Clark-type oxygen probe. sion in the test solution (Fig. 34.7), e.g. for field studies, allowing direct

measurement of oxygen status in situ, in contrast to chemical assays. The

main point to note is that the solution must be stirred during measure-
ment, to replenish the oxygen consumed by the electrode (‘boundary layer’
effect).
Definition
The glucose electrode
Biosensor – a device for measuring a The glucose electrode is a simple type of biosensor, whose basic design
substance, combining the selectivity of is shown in Fig. 34.8. It consists of a Pt electrode, overlaid by two mem-
a biological reaction with that of a sens-
branes. Sandwiched between these membranes is a layer of the immobi-
ing electrode.
lised enzyme glucose oxidase. The outer membrane is glucose permeable
and allows glucose in the sample to diffuse through to the glucose oxidase
layer, where it is converted to gluconic acid and H2O2. The inner membrane
is selectively permeable to H2O2, which is oxidised to O2 at the surface of
platinum the Pt electrode. The current arising from this release of electrons is pro-
electrode portional to the glucose concentration in the sample within the range 10-7
inner
to 10-3 mol L-1.
2e- + 2H2O + O2 H2O2 + 2OH- membrane Electrochemical detectors used in chromatography operate by voltam-
metric principles and currents are produced as the mobile phase flows over
glucose
electrodes set at a fixed potential: to achieve maximum sensitivity, this poten-
glucose
glucose + O2
oxidase
H2O2 + gluconic oxidase tial must be set at a level that allows electrochemical reactions to occur in all
acid (immobilised) analytes of interest.
outer
membrane Coulometric methods
glucose solution (sample) Here, the charge required to electrolyse a sample completely is measured: the
time required to titrate an analyte is measured at constant current and related
Fig. 34.8 Underlying principles of a glu- to the amount of analyte using Faraday’s law. There are few chemical applica-
cose electrode. tions of this technique, though it is sometimes used for determination of Cl-
in serum and body fluids.
Cyclic voltammetry
O R
The technique provides qualitative information about electrochemical reac-
tions, e.g. the redox behaviour of compounds and the kinetics of electron trans-
fer reactions. In practice, a triangular potential waveform is applied linearly to
cathodic
forward scan
the working electrode in a unstirred solution. After a few seconds, the ramp is
reversed and the potential is returned to its initial value. The process may be
current
repeated several times. The resulting plot of current versus potential is termed
a cyclic voltammogram. Fig. 34.9 shows a typical cyclic voltammogram for
anodic
reverse scan
a reversible redox couple after a single potential cycle. It is assumed that the
oxidised form, O, is the only species present at the start. Therefore, the first
scan is towards the (more) negative direction, commencing at a value were no
O R
reduction occurs. As the applied potential approaches the characteristic E o for
potential (V) the redox process, a catholic current starts to increase, up to a maximum. After
exceeding the potential at which the reduction process takes place, the direc-
Fig. 34.9 A typical cyclic voltammogram for
tion of the potential current is reversed. During this stage, reduced molecules
a reversible O + ne- 4 R redox process.
R, generated during the initial process, are reoxidised back to O, resulting in
an anodic peak.


Bard, A.J. and Faulkner, L.R. (2001) Electrochemical McMahon, G. (2008) Analytical Instrumentation: A Guide
Methods. 2nd edn. John Wiley & Sons Ltd, Chichester. to Laboratory, Portable and Miniaturized Instruments.
Hamann, C.H., Hamnett, A. and Vielstich, W. (2007) John Wiley & Sons Ltd, Chichester.
Electrochemistry, 2nd edn. John Wiley & Sons Ltd, Monk, P.M.S. (2001) Fundamentals of Electro-Analytical
Chichester. Chemistry. John Wiley & Sons Ltd, Chichester.
Kellner, R., Mermet, J.M., Otto, M., Valcarcel, M. and Schwedt, G. (1998) The Essential Guide to Analytical
Widmer, H.M. (2004) Analytical Chemistry: A Modern Chemistry. John Wiley & Sons Ltd, Chichester.
Approach to Analytical Science, 2nd edn. John Wiley & Wang, J. (2006) Analytical Electrochemistry, 3rd edn. John
Study exercises
34.1 Test your understanding of the circumstances equilibration at 101.3 kPa, what is the amount of
under which ion-selective electrodes can be oxygen in each of the following (give all answers
used. List the three main assumptions underly- to three significant figures):
ing the measurement of ion concentration using
(a) 4 mL of distilled water at 20°C (express your
an ion-selective electrode.
answer in mmol);
34.2 Test your knowledge of biosensor design. List
(b) 20 mL of sea water at 12°C (express your
the two main functional components of biosen-
answer in mmol);
sor such as the glucose electrode. Research a
(c) 10 mL of distilled water at 15°C (express your
specific application – note that it should be dif-
answer in mmol);
ferent from the ones used in this chapter (e.g.
(d) 250 mL of distilled water at 37°C (express
using the Web).
your answer in mg);
34.3 Calculate the oxygen content of specified vol-
(e) 200 mL of sea water at 25°C (express your
umes of water at defined temperatures. Using
answer in mL).
the information in Table 34.1 and assuming air

35 Using radioisotopes
The isotopes of a particular element have the same number of protons in the
Examples 126 C, 136 C and 146 C are three of nucleus but different numbers of neutrons, giving them the same proton number
the isotopes of carbon. About 98.9% (atomic number) but different nucleon numbers (mass number, i.e. number of
of naturally occurring carbon is in the
protons + number of neutrons). Isotopes may be stable or radioactive. Radi-
stable 126 C form. 136 C is also a stable
oactive isotopes (radioisotopes) disintegrate spontaneously at random to yield
isotope but it only occurs at 1.1% natu-
ral abundance. Trace amounts of radio- radiation and a decay product.
active 146 C are found naturally; this is a
negatron-emitting radioisotope. Radioactive decay
There are three forms of radioactivity (Table 35.1) arising from three main
types of nuclear decay:
●● Alpha decay. This involves the loss of a particle equivalent to a helium
Examples 226Ra decays to 222Rn by nucleus. Alpha (a) particles, being relatively large and positively charged,
loss of an alpha particle, as follows: do not penetrate far in living tissue, but they do cause ionisation damage and
226
88 Ra S 222 4
86 Rn + 2He
2+ this makes them generally unsuitable for tracer studies.
14
C shows beta decay, as follows: ●● Beta decay. This involves the loss or gain of an electron or its positive
14 S 147 N + b-
counterpart, the positron. There are three subtypes:
6C
22 (a) Negatron (b- ) emission: loss of an electron from the nucleus when
Na decays by positron emission, as
follows: a neutron transforms into a proton. This is the most important form
of decay for radioactive tracers used in chemistry. Negatron-emitting
22 S 22 +
11Na 10Ne + b isotopes of importance include 3H, 14C, 32P and 35S.
55
Fe decays by electron capture and (b) Positron (b+ ) emission: loss of a positron when a proton transforms into
the production of an X-ray, as follows: a neutron. This only occurs when sufficient energy is available from
55
26Fe S 55
25Mn + X the transition and may involve the production of gamma rays when the
The decay of 22Na by positron emis- positron is later annihilated by collision with an electron.
sion (b+) leads to the production of a (c) Electron capture (EC): when a proton ‘captures’ an electron and trans-
g ray when the positron is annihilated forms into a neutron. This may involve the production of X-rays as
on collision with an electron. electrons ‘shuffle’ about in the atom (as with 125I) and it frequently
involves electron emission.
●● Gamma emission. Internal transition involves the emission of electromag-
netic radiation in the form of gamma (g) rays from a nucleus in a metastable
state and always follows initial alpha or beta decay. Emission of gamma
radiation leads to no further change in atomic number or mass.
Note from the above that more than one type of radiation may be emitted
when a radioisotope decays. The main radioisotopes used in chemistry and
their properties are listed in Table 35.2.
Table 35.1 Types of radioactivity and their properties.
Range of maximum Penetration range Suitable shielding

Radiation energies (MeV*) in air (m) material
Alpha (a) 4–8 0.025–0.080 Unnecessary

Beta (b) 0.01–3 0.150–16 Plastic (e.g. Perspex)
Gamma (g) 0.03–3 1.3–13† Lead
*Note that 1 MeV = 1.6 * 10 -13 J.

† Distance at which radiation intensity is reduced to half.

Using radioisotopes
Table 35.2 Properties of some isotopes used commonly in the physical and life sciences. Physical data obtained from
Haynes (2014).
Maximum
Isotope Emission(s) energy (MeV) Half-life Main uses Advantages Disadvantages
-
3
H b 0.01861 12.3 years Suitable for labelling Relatively safe Low efficiency of
organic molecules in detection, high iso-
wide range of posi- tope effect, high rate
tions at high specific of exchange with
activity environment
14
C b- 0.156 48 5715 years Suitable for labelling Relatively safe, low Low specific activity
organic molecules in a rate of exchange with
wide range of positions environment
22
Na b+ (90%) + g, 2.842 (b+ ) 2.60 years Transport studies High specific activity Hazardous
EC
32
P b- 1.710 14.3 d Labelling proteins and High specific activity, Short half-life,
nucleotides (e.g. DNA) ease of detection hazardous
33
P b- 0.248 25 d Labelling nucleotides Safer than 32 P Moderate half-life
and proteins
35
S b- 0.167 87.2 d Labelling proteins and Low isotope effect Low specific activity
nucleotides
36
CI b- , b+ , EC 0.709 (b- ) 300 000 years Transport studies Low isotope effect Low specific activity,
1.142 (b+ , EC) hazardous
125
I EC + g 0.178 (EC) 59.9 d Labelling proteins and High specific activity Hazardous
nucleotides
131
I b- + g 0.971 (b- ) 8.04 d Labelling proteins and High specific activity Hazardous
nucleotides
EC, Electron capture.
Each radioactive particle or ray carries energy, usually measured in elec-

tron volts (eV). The particles or rays emitted by a particular radioisotope exhibit
a range of energies, termed an energy spectrum, characterised by the maximum
x energy of the radiation produced, E max. The energy spectrum of a particular
radioisotope is relevant to the following:
Radioactivity
●● Safety: isotopes with the highest maximum energies will have the great-
0.5x est penetrating power, requiring appropriate shielding (Table 35.1).
0.25x ●● Detection: different instruments vary in their ability to detect isotopes
with different energies.
Time Discrimination: some instruments can distinguish between isotopes,
t1/2 t1/2 ●●
based on the energy spectrum of the radiation produced (p. 315).

Fig. 35.1 Decay of a radioactive isotope with
time. The time taken for the radioactivity to The decay of an individual atom (a ‘disintegration’) occurs at random,
decline from x to 0.5x is the same as the but that of a population of atoms occurs in a predictable manner. The radi-
time taken for the radioactivity to decline oactivity decays exponentially, having a characteristic half-life (t1/2). This is
from 0.5x to 0.25x, and so on. This time is the time taken for the radioactivity to fall from a given value to half that value
the half-life (t1/2) of the isotope. (Fig. 35.1). The t1/2 values of different radioisotopes range from fractions of a
second to more than 1019 years. If t1/2 is very short, as with 15O (t1/2 ≈ 2 min),
then it is generally impractical to use the isotope in experiments because you
would need to account for the decay during the experiment and counting period.
To calculate the fraction (f) of the original radioactivity left after a particu-
lar time (t), use the following relationship:

Using radioisotopes
f = ex, where x = -0.693t/t1 [35.1]

Example For 35S, with a half-life 2
of 87.2 d, the f raction of radioactiv- Note that the same units must be used for t and t1/2 in this equation.
ity remaining after 28 d would be
worked out as follows: first, from eqn
[35.1], x = ( - 0.693 * 28) , 87.2 = Measuring radioactivity
- 0.222522936, then using eqn [35.1], The SI unit of radioactivity is the becquerel (Bq), equivalent to one disintegra-
f = e-0.222522936 = 0.800496646 (with
tion per second (d.p.s.), but disintegrations per minute (d.p.m.) are also used.
appropriate rounding, 80.0% of
original activity).
The curie (Ci) is a non-SI unit equivalent to the number of disintegrations pro-
duced by 1 g of radium (37 GBq). Table 35.2 shows the relationships between
these units. In practice, most instruments are not able to detect all of the disinte-
grations from a particular sample, i.e. their efficiency is less than 100% and the
rate of decay may be presented as counts min-1 (c.p.m.) or counts s-1 (c.p.s.).
Table 35.2 Relationships between units of Most modern instruments correct for background radiation and inefficiencies
radioactivity. For abbreviations, see text. in counting, converting count data to d.p.m. Alternatively, the results may be
presented as the measured count rate, although this is only valid where the
1 Bq = 1 d.p.s. 1 Sv = 100 rem efficiency of counting does not vary greatly among samples.
1 Bq = 60 d.p.m. 1 Gy = 100 rad
1 Bq = 27 pCi 1 Gy ≈ 100 roentgen
1 d.p.s. = 1 Bq
Key Point The specific activity is a measure of the quantity of
1 rem = 0.01 Sv
1 d.p.m. = 0.0167 Bq
radioactivity present in a known amount of the substance:
1 rad = 0.01 Gy
1 Ci = 37 GBq 1 roentgen ≈ 0.01 Gy radioactivity (Bq, Ci, d.p.m., etc.)
1 mCi = 37 MBq specific activity = [35.2]
amount of substance (mol, g, etc.)
1 mCi = 37 kBq
This is an important concept in practical work involving radioisotopes,

since it allows interconversion of disintegrations (activity) and amount of sub-
stance (see Box 35.1).
Two SI units refer to doses of radioactivity and these are used when cal-
Example If 0.4 mL of a 32P@labelled culating exposure levels for a particular source. The sievert (Sv) is the amount
DNA solution at a concentration of
of radioactivity giving a dose in humans equivalent to 1 gray (Gy) of X-rays:
50 mmol L-1 (amount = 0.4 * 50 ,
1 Gy = an energy absorption of 1 J kg 1. The dose received in most biological
1000 = 0.02 mmol) gave a count
of 2490 d.p.m. ( = 41.5 Bq), experiments is a negligible fraction of the maximum permitted exposure limit.
using eqn [35.2] this would cor- Conversion factors from older units are given in Table 35.2.
respond to a specific activity of The most important methods of measuring radioactivity for biological pur-
2490 , 0.02 = 124 500 d.p.m. mmol-1 poses are described below.
(or 2075 Bq mmol-1).
The Geiger–Müller (G–M) tube
This operates by detecting radiation when it ionises gas between a pair of elec-
cathode trodes across which a voltage has been applied (Fig. 35.2). You should use a
anode handheld Geiger–Müller tube for routine checking for contamination (although
it will not pick up 3 H activity).
The scintillation counter

This operates by detecting the scintillations (fluorescence ‘flashes’) produced
gas at low thin mica when radiation interacts with certain chemicals called fluors (Fig. 35.3). In
pressure window
solid (or external) scintillation counters (often referred to as ‘gamma counters’)
the radioactivity causes scintillations in a crystal of fluorescent material held
0 2 1 ‘counter’ close to the sample. This method is only suitable for radioisotopes producing
penetrating radiation.
Liquid scintillation counters are mainly used for detecting beta decay
Fig. 35.2 Components of a Geiger–Müller and they are especially useful in biology. The sample is dispersed or dis-
(G–M) tube. solved in a suitable solvent containing the fluor(s) – the ‘scintillation

Using radioisotopes
Box 35.1 How to determine the specific activity of an experimental solution
Suppose you need to make up a certain volume of an 6. Measure the radioactivity in a known volume of the
experimental solution, to contain a particular amount of experimental solution. If you are using an instru-
radioactivity. For example, 50 mL of a mannitol solution ment with automatic correction to Bq, your sample
at a concentration of 25 mmol L-1, to contain 5 Bq mL-1 – should contain the predicted amount of radioactivity,
using a manufacturer’s stock solution of 14C@labelled e.g. an accurately dispensed volume of 100 mL of the
mannitol (specific activity = 0.1 Ci mmol-1). mannitol solution should give a corrected count of
100 * 5 = 500 Bq (or 500 * 60 = 30 000 d.p.m.).
1. Calculate the total amount of radioactivity in the
experimental solution, in this example 5 * 1000 7. Note the specific activity of the experimental solu-
(to convert mL to mL) * 50 (50 mL required) tion: in this case, 100 mL (1 * 10-4 l) of the manni-
= 2.5 * 105 Bq (i.e. 250 kBq). tol solution at a concentration of 0.025 mol L-1 will
contain 25 * 10-7 mol (2.5 mmoL) mannitol. Dividing
2. Establish the volume of stock radioisotope solution
the radioactivity in this volume (30 000 d.p.m.) by
required: for example, a manufacturer’s stock solution
the amount of substance (eqn [35.2]) gives a spe-
of 14C@labelled mannitol contains 50 mCi of radioisotope
cific activity of 30 000/2.5 = 12 000 d.p.m. mmol-1, or
in 1 mL of 90% v/v ethanol: water. Using Table 35.2, this
12 d.p.m. nmol-1. This value can be used:
is equivalent to an activity of 50 * 37 = 1850 kBq. So,
the volume of solution required is 250/1850 of the stock (a) To assess the accuracy of your protocol for pre-
volume, i.e. 0.1351 mL (135 mL). paring the experimental solution: if the meas-
ured activity is substantially different from the
3. Calculate the amount of non-radioactive
predicted value, you may have made an error in
s ubstance required as for any calculation
making up the solution.
involving concentration (see p. 445), e.g. 50 mL
(0.05 L) of a 25 mmol L-1(0.025 mol L-1) manni- (b) To determine the counting efficiency of an instru-
tol (relative molecular mass 182.17) will contain ment; by comparing the measured count rate with
0.05 * 0.025 * 182.17 = 0.2277 g. the value predicted by your calculations.
4. Check the amount of radioactive isotope to be added. In (c) To interconvert activity and amount of substance:
most cases, this represents a negligible amount of sub- the most important practical application of spe-
stance, e.g. in this instance, 250 kBq of stock solution cific activity is the conversion of experimental
at a specific activity of 14.8 * 106 kBq mmol-1 (con- data from counts (activity) into amounts of sub-
verted from 0.4 Ci mmol-1 using Table 45.3) is equal to stance. This is only possible where the substance
250/14 800 000 = 16.89 nmol, equivalent to approxi- has not been metabolised or otherwise converted
mately 3 mg mannitol. This can be ignored in calculating into another form; e.g. a sample incubated in
the mannitol concentration of the experimental solution. the experimental solution described above with
a measured activity of 245 d.p.m. can be con-
5. Make up the experimental solution by adding the
verted to nmol mannitol by dividing by the spe-
appropriate amount of non-radioactive substance and
cific activity, expressed in the correct form. Thus
the correct volume of stock solution.
245/12 = 20.417 nmol mannitol.
photomultiplier tubes (2) cocktail’. The radiation first interacts with the solvent, and the energy from
pulse analysis
lead shielding and readout
this interaction is passed to the fluors, which produce detectable light. The
scintillations are measured by photomultiplier tubes which turn the light
pulses into electronic pulses, the magnitude of which is directly related
1 4 5
to the energy of the original radioactive event. The spectrum of electronic
pulses is thus related to the energy spectrum of the radioisotope.
sample vial Modern liquid scintillation counters use a series of electronic ‘windows’
to split the pulse spectrum into two or three components. This may allow more
than one isotope to be detected in a single sample, provided their energy spectra
sample, dispersed or dissolved in ‘scintillation cocktail’
are sufficiently different (Fig. 35.4). A complication of this approach is that
Fig. 35.3 Components of a scintillation the energy spectrum can be altered by pigments and chemicals in the sam-
counter. Note that in most modern instru- ple, which absorb scintillations or interfere with the transfer of energy to the
ments, all components are enclosed within fluor; this is known as quenching (Fig. 35.4). Most instruments have computer-
a single cabinet. operated quench correction facilities (based on measurements of standards of

Using radioisotopes
‘window b’ ‘window a’
b’
b
Frequency
a
Emax
Energy
Fig. 35.4 Energy spectra for three radioactive samples, detected using a
scintillation counter. Sample a is a high energy b@emitter while b contains
a low energy b@emitter, giving a lower spectral range. Sample b’ contains
the same amount of low energy b@emitter, but with quenching, shifting the
spectral distribution to a lower energy band. The counter can be set up to
record disintegrations within a selected range (a ‘window’). Here, ‘window
a’ could be used to count isotope a while ‘window b’ could give a value of
isotope b, by applying a correction for the counts due to isotope a, based
on the results from ‘window a’. Dual counting allows experiments to be
carried out using two isotopes (double labelling).
known activity and energy spectrum) which correct for such changes in count-
Correcting for quenching – find out
ing efficiency.
how your instrument corrects for
Many liquid scintillation counters treat the first sample as a ‘background’,
quenching and check the quench indi-
subtracting whatever value is obtained from the subsequent measurements as
cation parameter (QIP) on the printout,
part of the procedure for converting to d.p.m. If not, you will need to subtract
which measures the extent of quench-
the background count from all other samples. Make sure that you use an appro-
ing of each sample. Large differences in
priate background sample, identical in all respects to your radioactive sample
the QIP would indicate that quenching
but with no added radioisotope, in the correct position within the machine.
is variable among samples and might
Check that the background reading is reasonable (15–30 c.p.m. is a reasona-
give you cause for concern.
ble background for most radioisotopes). Tips for preparing samples for liquid
scintillation counting are given in Box 35.2.
Liquid scintillation counting of high Gamma-ray (G@ray) spectrometry

energy B@emitters – b@particles with This is a method by which a mixture of g@ray@emitting radionuclides can be
energies greater than 1 MeV can be resolved quantitatively by pulse-height analysis. It is based on the fact that pulse
counted in water (Cerencov radiation), heights (voltages) produced by a photomultiplier tube are proportional to the
with no requirement for additional amounts of g@ray energy arriving at the scintillant or a lithium-drifted germa-
fluors (e.g. 32P). nium detector. The lithium-drifted germanium detector, which is abbreviated to
Ge(Li) – pronounced ‘jelly’ – provides high resolution (narrow peaks), essential
in the analysis of complex mixtures, such as in environmental material.
Autoradiography
This is a method where photographic film is exposed to the isotope. It is used
mainly to locate radioactive tracers in thin sections of an organism or on chro-
matography papers and gels, but quantitative work is possible. The radiation
interacts with the film in a similar way to light, silver grains being formed in the
developed film where the particles or rays have passed through. The radiation

Using radioisotopes
Box 35.2 Tips for preparing samples for liquid scintillation counting
Modern scintillation counters are very simple to operate; (c) Particulate samples may sediment to the bottom of
problems are more likely to be caused by inadequate the scintillation vial – suspend them by forming a
sample preparation than to incorrect operation of the gel. This can be done with certain emulsifier-based
machine. Common pitfalls are the following: cocktails by adding a specific amount of water.
●● Incomplete dispersal of the radioactive compound in ●● Chemiluminescence. This is where a chemical reacts
the scintillation cocktail. This may lead to underesti- with the fluors in the scintillation cocktail, causing
mation of the true amount of radioactivity present: spurious scintillations, a particular risk with solutions
containing strong bases or oxidising agents. Symp-
(a) Water-based samples may not mix with the scintil- toms include very high initial counts which decrease
lation cocktail – change to an emulsifier-based cock- through time. Possible remedies are:
tail. Take care to observe the recommended limits,
upper and lower, for amounts of water to be added (a) Leave the vials at room temperature for a time
or the cocktail may not emulsify properly. before counting. Check with a suitable blank that
(b) Solid specimens may absorb disintegrations or counts have dropped to an acceptable level.
scintillations: extract radiochemicals using an (b) Neutralise basic samples with acid (e.g. acetic acid
intermediate solvent like ethanol (ideally within or HCI).
the scintillation vial) and then add the cocktail. (c) Use a scintillation cocktail that resists chemilumi-
Tissue-solubilising compounds such as Soluene nescence, such as Hiconicfluor.
are effective, particularly for animal material, but
(d) Raise the energy of the lower counts detected to
extremely toxic, so the manufacturer’s instructions
about 8 keV – most chemiluminescence pulses are
must be followed closely. Radioactive compounds
weak (0–7 keV). This approach is not suitable for
on slices of agarose or poly-acrylamide gels may
weak emitters, e.g. 3 H.
be extracted using a product such as Protosol. Aga-
rose gels can be dissolved in a small volume of
boiling water.
must have enough energy to penetrate into the film, but if it has too much energy
the grain formation may be too distant from the point where the isotope was
located to identify precisely the point of origin (e.g. high energy b@emitters).
Autoradiography is a relatively specialised method and individual lab protocols
should be followed for particular isotopes/applications.
Chemical applications for radioactive isotopes

The main advantages of using radioactive isotopes in chemical experiments are:
●● Radioactivity is readily detected. Methods of detection are sufficiently
sensitive to measure extremely small amounts of radioactive substances.
●● Studies can be carried out on intact, living organisms. If care is taken,
minimal disruption of normal conditions will occur when radiolabelled com-
pounds are introduced.
●● Protocols are simple compared to equivalent methods for chemical analysis.
The main disadvantages are:
●● The ‘isotope effect’. Molecules containing different isotopes of the same atom
may react at slightly different rates and behave in slightly different ways from
the natural isotope. The isotope effect is more extreme the smaller the atom,
and is most important for 3H@labelled compounds of low molecular mass.
●● The possibility of mistaken identity. The presence of radioactivity does not
tell you anything about the compound in which the radioactivity is present:

Using radioisotopes
it could be different from the one in which it was applied, due to metabolism
Investigating the metabolic fate of
or spontaneous breakdown of a 14C@containing organic compound.
radiolabelled compounds – you may
need to separate individual metabolites The main types of experiments are:
before counting, e.g. using chromatog-
●● Radiolabelled compounds: the use of radiolabelled compounds in synthetic
raphy (Chapter 32), or electrophoresis
and tracer studies in important as it allows the scientist to locate the labelled
(Chapter 33).
atom, i.e. 1 4 C, 3 H, in, for example, chemical synthesis and laboratory envi-
ronmental fate (degradation) studies. If using radiolabelled compounds sev-
eral issues arise and these include deciding upon the radionuclide itself, its
position in the molecule, the specific activity, the solvent and cost.
●● Radio-dating: the age of plant or mineral samples can be determined by
Example Carbon dating – living measuring the amount of a radioisotope in the sample. The age of the specimen
organisms have essentially the same can be found using t1/2 by assuming how much was originally incorporated.
ratio of 14C to 12C as the atmosphere;
however, when an organism dies, its ●● Medical uses: in radiotherapy the use of gamma radiation from 60Co to
14 12
C/ C falls because the radioactive destroy cancerous cells; 24Na can be introduced into the blood stream to
14
C isotope decays. Since we know the follow the flow of blood and identify obstructions; heart disease can be
half-life of 14C (5715 years), a sample’s assessed using 201Tl and 99Tc where the former concentrates in healthy heart
14 12
C/ C ratio will allow us to estimate tissue and the latter concentrates in abnormal heart tissue.
its age; e.g. if the ratio were exactly 1/8
of that in the atmosphere, the sample ●● Assays: radioisotopes are used in several quantitative detection methods of
is three half-lives old and was formed value to chemists. Radioimmunoassay is a quantitative method for meas-
17 145 years before present. Such urement of a substance (the analyte) using antibodies which bind specif-
estimates carry an error of the order ically to that analyte. Isotope dilution analysis works on the assumption
of 10% and are unreliable for samples that introduced radiolabelled molecules will equilibrate with unlabelled
older than 50 000 years, for which
molecules present in the sample. The amount of substance initially present
longer-lived isotopes can be used.
can be worked out from the change in specific activity of the radioisotope
when it is diluted by the ’cold’ material. A method is required whereby the
substance can be purified from the sample and sufficient substance must be
present for its mass to be measured accurately. Activation analysis is a sen-
sitive technique for the determination of element concentration. It is based
upon selectivity inducing radioactivity in some of the atoms of the elements
comprising the sample and then selectivity measuring the radiations emitted
by the radionuclides. After bombardment with suitable nuclear particles, the
induced radionuclides are identified or quantitatively measured. Neutron
activation analysis is the most common method of analysis.
Lowenthal and Airey (2005) give further details and applications.
Working practices when using radioactive isotopes

Registering for radioisotope work – in By law, undergraduate work with radioactive isotopes must be very closely
the UK, institutions must be registered supervised. In practical classes, the protocols will be clearly outlined, but in
for work with specific radioisotopes project work you may have the opportunity to plan and carry out your own
under the Radioactive Substances Act experiments, albeit under supervision. Some of the factors that you should take
(1993). into account, based on the assumption that your department and laboratory are
registered for radioisotope use, are discussed below:
1. Must you use radioactivity? If not, it may be a legal requirement that
Supervision of work with radioiso- you use an alternative method.
topes – in the UK, the Ionising Radia-
2. Have you registered for radioactive work? Usually, all users must regis-
tions Act (1985) provides details of local
ter with a local Radiation Protection Supervisor. Details of the project may
arrangements for the supervision of
have to be approved by the appropriate administrator(s). You may have to
radioisotope work.
have a short medical examination before you can start work.

Using radioisotopes
3. What labelled compound will you use? Radioactive isotopes must be

Safety Note Planning radioi- ordered well in advance through your department’s Radiation Protection
sotope work – each new experiment Supervisor. Aspects that need to be considered include:
should be planned carefully and exper-
imental protocols laid down in advance (a) The radionuclide. With many organic compounds this will be confined
so you work as safely as possible and to 3 H and 1 4 C. The involvement of a significant ‘isotope effect’ may
do not waste expensive radioactively influence this decision (see p. 315).
labelled compounds. (b) The labelling position. This may be a crucial part of a metabolic study.
Specifically labelled compounds are normally more expensive than
those that are uniformly (‘generally’) labelled.
(c) The specific activity. The upper limit for this is defined by the iso-
tope’s half-life but, below this, the higher the specific activity, the
more expensive the compound.
4. Are suitable facilities available? You will need a suitable work area,
preferably out of the way of general lab traffic and within a fume cupboard
for those cases where volatile radioactive substances are used or may be
produced.
In conjunction with your supervisor, decide whether your method of
Carrying out a ‘dry run’ – consider application will introduce enough radioactivity into the system, how you will
doing this before working with radioac- account for any loss of radioactivity during recovery of the isotope and whether
tive compounds, perhaps using a dye there will be enough activity to count at the end. You should be able to predict
to show the movement or dilution of approximately the amount of radioactivity in your samples, based on the spe-
introduced liquids, as this will lessen cific activity of the isotope used, the expected rate of uptake/exchange and the
the risks of accident and improve your amount of sample to be counted. Use the isotope’s specific activity to estimate
technique. whether the non-radioactive (‘cold’) compound introduced with the radiola-
belled (‘hot’) compound may lead to excessive concentrations being adminis-
tered. Advice for handling data is given in Box 35.1.
Safety and procedural aspects
Safety Note Using Benchkote –
the correct way to use Benchkote and Make sure the bench surface is one that can be easily decontaminated by
similar products is with the waxed sur- washing (e.g. Formica) and always use a disposable surfacing material such
face down (to protect the bench or tray as Benchkote. It is good practice to carry out as many operations as possible
surface) and the absorbent surface up within a Benchkote-lined plastic tray so that any spillages are contained. You
(to absorb any spillage). Write the date will need a lab coat to be used exclusively for work with radioactivity, safety
in the corner when you put down a spectacles and a supply of thin latex or vinyl disposable gloves. Suitable
new piece. Monitor using a G–M tube vessels for liquid waste disposal will be required and special plastic bags
and replace regularly under normal cir- for solids – make sure you know beforehand the disposal procedures for
cumstances. If you are aware of spill- liquid and solid wastes. Wash your hands after handling a vessel containing
age, replace immediately and dispose a radioactive solution and again before removing your gloves. Gloves should
of correctly. be placed in the appropriate disposal bag as soon as your experimental pro-
cedures are complete.
It is important to comply with the following guidelines:
●● Read and obey the local rules for safe usage of radiochemicals.
●● Maximise the distance between you and the source as much as possible.
●● Minimise the duration of exposure.
●● Wear protective clothing (properly fastened lab coat, safety glasses, gloves)
at all times.
●● Use appropriate shielding at all times (Table 35.1).
●● Monitor your working area for contamination frequently.

Using radioisotopes
●● Mark all glassware, trays, bench work areas, etc., with tape incorporating
the international symbol for radioactivity (Fig. 35.5).
CAUTION
RADIOACTIVE ●● Keep adequate records of what you have done with a radioisotope – the
MATERIAL
amount remaining and that disposed of in waste form must agree.
●● Store radiolabelled compounds appropriately and return them to storage
Fig. 35.5 Tape showing the international immediately after use.
symbol for radioactivity.
●● Dispose of waste promptly and with due regard for local rules.
●● Make the necessary reports about waste disposal, etc., to your departmen-
tal Radiation Protection Supervisor.
●● Clear up after you have finished each experiment.
●● Wash thoroughly after using radioactivity.
●● Monitor the work area and your body when finished.

L’Annunziata, M.F. (ed.) (2012) Handbook of Radioactivity Lowenthal, G. and Airey, P. (2005) Practical Applications
Analysis, 3rd edn. Academic Press, San Diego. of Radioisotopes and Nuclear Radiations. Cambridge
Haynes, W.M. (ed.) (2014) CRC Handbook of University Press, Cambridge.
Chemistry and Physics, 95th edn. CRC Press, Boca Slater, J. (ed.) (2002) Radioisotopes in Biology: a Practical
Raton. Approach, 2nd edn. Oxford University Press: Oxford.
Study exercises
35.1 Carry out a half-life calculation. A rat dropping 35.3 Use the concept of specific activity in calculations.
found in a pyramid in Egypt had a 14C: 12C ratio A researcher wishes to estimate the rate of
that was 57.25% of a modern-day standard. Use uptake of the sugar galactose by carrot cells in
this value to estimate the approximate date when a suspension culture. She prepares 250 mL of
the rat visited the pyramid, to the nearest century. the cell culture medium containing 107 cells per
35.2 Practise radioactivity interconversions. Express mL and unlabelled galactose at a concentration
the following values in the alternative units indi- of 5 mmol L-1. She then ‘spikes’ this with 5 mL
cated, with appropriate prefixes as necessary. (regard this as an insignificant volume) of radio-
Answer to three significant figures. active standard containing 55 MBq of 14C@labelled
galactose (regard as an insignificant concentra-
(a) 72 000 d.p.m. as Bq;
tion). Answer to two significant figures.
(b) 20 mCi as d.p.m.;
(c) 44 400 Bq as mCi; (a) Calculate the specific activity of the galactose
(d) 6.3 * 105 d.p.m. mol-1 as Bq g -1, for a com- in the culture solution in Bq mol-1.
pound with a relative molecular mass of 350; (b) If the total cell sample takes up 79.2 * 105 Bq
(e) 3108 d.p.m. as pmol, for a sample of a stand- in a 2-h period, calculate the galactose uptake
ard where the specific activity is stated as rate in mol s-1 cell-1.
50 Ci mol-1.

36 Infrared and Raman spectroscopy
In addition to ultraviolet–visible (UV–vis) spectroscopy (p. 217), there are

Identifying compounds – the combi-
three other essential techniques that you will encounter during your laboratory
nation of techniques described in this
course. They are:
and the following chapters can often
provide sufficient information to iden- 1. Infrared (IR) spectroscopy: this is concerned with the energy changes
tify a compound with a low probability involved in the stretching and bending of covalent bonds in molecules.
of error. 2. Nuclear magnetic resonance (NMR) spectroscopy: this involves the
absorption of energy by specific atomic nuclei in magnetic fields and is
probably the most powerful tool available for the structural determination
Definitions of molecules (Chapter 37).
3. Mass spectrometry (MS): this is based on the fragmentation of compounds
Spectroscopy – any technique involving
into smaller units. The resulting positive ions are then separated according
the production and subsequent record-
to their mass-to-charge ratio (m/z) (Chapter 38).
ing of a spectrum of electromagnetic
radiation, usually in terms of wave- As with UV–vis spectroscopy, IR and NMR spectroscopy are based on the
length or energy. interaction of electromagnetic radiation with molecules, whereas MS is differ-
Spectrometry – any technique involving ent in that it relies on high-energy particles (electrons or ions) to break up the
the measurement of a spectrum, e.g. of molecules. The relationship between the various types of spectroscopy and the
electromagnetic radiation, molecular electromagnetic spectrum (see p. 217) is shown in Table 36.1.
masses.
Infrared spectroscopy
A covalent bond between two atoms can be crudely modelled as a spring con-
Interpreting spectra – the spectrum necting two masses and the frequency of vibration of the spring is defined
produced in UV–vis, IR and NMR spec- by Hooke’s law (eqn [36.1]), which relates the frequency of the vibration (n)
troscopy is a plot of wavelength or to the strength of the spring, expressed as the force constant (k), and to the
frequency or energy (x-axis) against masses (m1 and m2) on the ends of the spring [defined as the reduced mass
absorption of energy (y-axis). Conven- m = (m1 * m2) , (m1 + m2)].
tion puts high frequency (high energy,
short wavelength) at the left-hand side 1 k
n = [36.1]
of the spectrum. 2p A m
In simple terms, this means that:

●● the stretching vibration of a bond between two atoms will increase in fre-
quency (energy) if on changing from a single bond to a double bond and
Table 36.1 The electromagnetic spectrum and types of spectroscopy
Type of radiation Origin Wavelength Type of spectroscopy
g@rays Atomic nuclei 6 0.1 nm g@ray spectroscopy

X-rays Inner shell electrons 0.01–2.0 nm X-ray fluorescence (XRF)
Ultraviolet (UV) Ionisation 2.0–200 nm Vacuum UV spectroscopy
UV/visible Valency electrons 200–800 nm UV/visible spectroscopy
Infrared Molecular vibrations 0.89300 mm IR and Raman spectroscopy
Microwaves Molecular rotations 1 mm to 30 cm Microwave spectroscopy
Electron spin Electron spin resonance (ESR)
Radio waves Nuclear spin 0.6–10 m Nuclear magnetic resonance (NMR)

Infrared and Raman spectroscopy
then to a triple bond between the same two atoms (masses), i.e. the spring
gets stronger. For example,
n for C ‚ C 7 n for C “ C 7 n for C ¬ C
●● as the masses of the atoms on a bond increases, the frequency of the vibration
decreases, i.e the effect of reducing the magnitude of m; for example,
n for C ¬ H 7 n for C ¬ C; n for C ¬ H 7 n for C ¬ D;

n for O ¬ H 7 n for S ¬ H
Bonds can also bend, but this movement requires less energy than stretch-
ing and thus the bending frequency of a bond is always lower than the corre-
sponding stretching frequency. When IR radiation of the same frequency as
the bond interacts with the bond it is absorbed and increases the amplitude of
vibration of the bond. This absorption is detected by the IR spectrometer and
results in a peak in the spectrum. For a vibration to be detected in the IR region
IR absorption bands – since the fre-
the bond must undergo a change in dipole moment when the vibration occurs.
quency of vibration of a bond is a spe-
Bonds with the greatest change in dipole moment during vibration show the
cific value you would expect to see line
most intense absorption, e.g. C “ O and C ¬ O.
spectra on the chart. However, each
Since bonds between specific atoms have particular frequencies of vibra-
vibration is associated with several
tion, IR spectroscopy provides a means of identifying the type of bonds in a
rotational motions and bands (peaks)
molecule, e.g. all alcohols will have an O ¬ H stretching frequency and all
are seen in the spectrum.
compounds containing a carbonyl group will have a C “ O stretching fre-
quency. This property, which does not rely on chemical tests, is extremely
useful in diagnosing the functional groups within a covalent molecule.
IR spectra
A typical IR spectrum is shown in Fig. 36.1 and you should note the following
points:
●● The x-axis, the wavelength of the radiation, is given in wavenumbers (n)
and expressed in reciprocal centimetres (cm -1). You may still see some
%T
100.00
1447.8 –
2984.9 –
1373.9 –
1047.3 –
1241.0 –
1741.9 –
0.00
4000 3500 3000 2500 2000 1500 1000 600 cm-1
Fig. 36.1 IR spectrum of ethyl ethanoate CH3COOCH2CH3 as a liquid film.

spectra from old instruments using microns (m, equivalent to the SI unit
The use of wavenumber – this is an
‘micrometres’, mm, at 1 * 10-6 m) for wavelength; the conversion is given
old established convention, since high
by eqn [36.2]:
wavenumber = high frequency = high
energy = short wavelength. Expression wavenumber (cm -1) = 1/wavelength (cm) = 10 000/wavelength (mm)
of the IR range, 4000 cm -1 to 650 cm -1, [36.2]
is in ‘easy’ numbers and the high
energy is found on the left-hand side ●● The y-axis, expressing the amount of radiation absorbed by the molecule, is
of the spectrum. Note that IR spectros- usually shown as % transmittance (p. 218). When no radiation is absorbed
copists often refer to wavenumbers as (all is transmitted through the sample) we have 100% transmittance and 0%
‘frequencies’, e.g. ‘the peak of the C “ O transmittance implies all radiation is absorbed at a particular wavenumber.
stretching “frequency” is at 1720 cm -1’. Since the y-axis scale goes from 0 to 100% transmittance, the absorption
peaks are displayed down from the 100% line; this is opposite to most other
common spectra.
●● The cells holding the sample usually display imperfections and are not
completely transparent to IR radiation, even when empty. Therefore the
base line of the spectrum is rarely set on 100% transmittance and quanti-
tative applications of IR spectroscopy are more complex than for UV–vis
(p. 220).
motor
IR spectrometers
amplifier recorder There are two general types:
1. Double-beam or dispersive instruments in which the IR radiation from a
detector wedge
monochromator single source is split into two identical beams. One beam passes through
chopper source the sample and the other is used as a reference and passes through air or the
slits
sample pure solvent used to dissolve the sample. The difference in intensity of the
beam
grating two beams is detected and recorded as a peak; the principal components
of this type of instrument are shown in Fig. 36.2. The important controls
Fig. 36.2 Schematic diagram of a d
ouble- on the spectrometer are:
beam IR spectrometer.
(a) scan speed: this is the rate at which the chart moves – slower for
greater accuracy and sharp peaks;
Using double-beam instruments – you (b) wavelength range: the full spectrum or a part of the IR range may be
can identify the sample beam by quickly selected;
placing your hand in the beam. If the
(c) 100% control: this is used to set the pen at the 100% transmittance line
pen records a peak, this is the sample
when no sample is present the base line. It is usual practice to set the
beam, but if the pen moves up, then this
pen at 90% transmittance at 4000 cm -1 when the sample is present,
is the reference beam.
to give peaks of the maximum deflection.
You should remember that this is an electromechanical instrument and you
should always make sure that you align the chart against the calibration
Using the 100% control – if you use marks on the chart holder. In the more advanced instruments an on-board
this control to set the base line for the computer stores a library of standard spectra, which can be compared with
sample, you must turn down the 100% your experimental spectrum.
control when you remove the sample,
otherwise the pen-drive mechanism 2. Fourier transform IR (FT–IR) spectrometer: the value of IR spectroscopy
may be damaged in trying to drive off is greatly enhanced by Fourier transformation, named after the mathema-
the top of the chart. tician J.B. Fourier. The FT is a procedure for inter-converting frequency
functions and time or distance functions. The IR beam, composed of all
the frequencies in the IR range, is passed through the sample and generates
interference patterns, which are then transformed electronically into a nor-
mal IR spectrum A simple schematic diagram of an FT–IR spectrometer
is shown in Fig. 36.3. The advantages of FT–IR are:

Fixed mirror (a) rapid scanning speed – typically four scans can be made per minute,
Beam splitter Moving mirror
allowing addition of the separate scans to enhance the signal-to-noise
ratio and improve the resolution of the spectrum;
Source Collimator
(b) simplicity of operation – the reference is scanned first, stored and then
Sample subtracted from the sample spectrum;
compartment
(c) enhanced sensitivity: the facility of spectrum addition from multiple
Detector scans permits detection of smaller quantities of chemicals;
Fig. 36.3 Schematic: diagram of an FTIR (d) the integral computer system enables the use of libraries of spectra
spectrometer and simplifies spectrum manipulation, such as the subtraction of con-
taminant or solvent spectra.
The procedures for running IR spectra on double-beam and FT spectrometers
are described in Box 36.1.
Box 36.1 How to run an infrared spectrum of a liquid, solid film, mull or KBr disk
A. Double-beam spectrometer 2. Select the number of scans; usually four is adequate

for routine work.
1. Ensure that the instrument is switched on and that it
has had a few minutes to warm up. 3. Select ‘background’ on the on-screen menu, and scan
the background. Do not press any other buttons or
2. Make sure that the chart is aligned with the calibra-
icons while the spectrum is running.
tion marks on the chart bed or chart drum. Most spec-
trometers scan from 4000 cm -1 to 650 cm -1 and the 4. Place the sample cell in the beam, close the lid,
pen should be at the 4000 cm -1 mark. select ‘sample’ and scan the sample. Do not press
any other buttons or icons while the spectrum is
3. Adjust the 100% transmittance control to about 90%,
running.
if necessary.
5. Select ‘customise’, or a similar function, and enter all
4. Place the sample cell in the sample beam and adjust
the data – name, date, compound, phase (liquid film,
the 100% transmittance control to 90%, or the highest
Nujol® mull, KBr disk, etc.) – on the spectrum.
value possible.
6. Select ‘print’, to produce the spectrum from the
5. Select the scan speed. You must balance the defini-
printer.
tion required in the spectrum with the time available
for the experiment. For most qualitative applications Problems with IR spectra
the fastest setting is satisfactory.
These are usually caused by poor sample preparation
6. Press the ‘scan’ or ‘start’ button to run the spectrum. and the more common faults are:
The spectrum will be recorded and the spectrometer
1. The large peaks have tips below the bottom of the
will automatically align itself at the end of the run.
chart or the large peaks have ‘squared tips’ near the
Do not press any other buttons while the spectrum is
bottom of the chart: the sample is too thick; remove
running or the instrument may not realign itself at the
some sample from the cell and rerun the spectrum.
end of the run.
2. The spectrum is ‘weak’, i.e. few peaks: the sample is
7. Adjust the 100% transmittance control to about 50%,
too thin – add more sample or remake the KBr disk.
remove the sample cell from the spectrometer and
turn the 100% transmittance control to about 90%. 3. The base line cannot be adjusted to 90% trans-
mittance: the NaCl plates or KBr disk are ‘fogged’,
8. Enter all of the following data on the spectrum: name,
scratched or dirty – replace or remake the KBr disk.
date, compound and phase (liquid film, Nujol® mull,
KBr disk, etc.). 4. The pen tries to ‘go off’ the top of the spectrum:
obviously due to some absorption at 4000 cm -1
B. FT–IR spectrometer
when you were setting the base line. Repeat base-
1. Make sure that the sample compartment is empty line set-up but at 80% transmittance and bear in
and close the lid. mind that dirty plates, above, can be the cause.

gaskets Sample handling

You can obtain IR spectra of solids, liquids and gases by use of the appropriate
sample cell (sample holder). The sample holder must be completely transparent
to IR radiation; consequently glass and plastic cells cannot be used. The most
common sample cells you will encounter are made from sodium chloride or
NaCl plates
potassium bromide and you cannot use aqueous solutions or very wet sam-
ples, otherwise the sample cells will dissolve. A typical range of sample cells
(a) is shown in Fig. 36.4 and for routine qualitative work you will regularly use
NaCl plates and KBr disks to obtain spectra of solids and liquids. Solution cells
and gas cells are utilised in more specialised applications and require specific
NaCl instructions and training.
window
Liquid samples
The most convenient way to obtain the IR spectrum of a pure, dry liquid is to
(b) make a thin liquid film between two NaCl disks (plates). Since the film thick-
ness is unknown, this procedure is not applicable to quantitative work.
Solid samples
If you were to place a fine powder between two NaCl plates, a usable spectrum
would not be obtained because the IR radiation would be scattered by diffrac-
NaCl tion at the edges of the particles and would not pass through to the detector.
window
There are three solutions to this problem:
1. Mulls: in which the finely ground solid is mixed with a liquid, usu-
ally Nujol® (liquid paraffin) or, less frequently, HCB (hexachloro-1,3-
butadiene). This mulling liquid does not dissolve the chemical but fills the
(c)
gaps round the edges of the crystals preventing diffraction and scattering
Fig. 36.4 Cells for IR spectroscopy: (a) of the IR radiation. Remember that these mulling liquids have their own
demountable cell for liquid and solid films IR spectrum, which is relatively simple, and can be subtracted either ‘men-
and mulls; (b) solution cell; (c) gas cell. tally’ or by the computer. The choice of mulling liquid depends upon the
region of the IR spectrum of interest: Nujol® is a simple hydrocarbon con-
taining only C9H and C ¬ C bonds, whereas HCB has no C ¬ H bonds,
IR spectra of aqueous solutions – but has C ¬ Cl, C “ C and C ¬ C bonds. Examination of the separate
special sample cells made from CaF2 spectra of your unknown compound in each of these mulling agents ena-
are available for aqueous solutions, bles the full spectrum to be analysed.
but they are expensive and only used
2. KBr disks: here the finely ground solid compound is mixed with anhydrous
in specific applications.
KBr and squeezed under pressure. The KBr becomes fluid and forms a
disk containing the solid compound dispersed evenly within it and suitable
for obtaining a spectrum. The advantage of the KBr disk technique is the
Storing IR sample cells and KBr powder – absence of the spectrum from the mulling liquid, but the disadvantages are
NaCl cells are always stored in desic- the equipment required (Fig. 36.5) and the practice required to obtain suit-
cators to prevent ‘fogging’ by absorp- able transparent disks, which are very delicate and rapidly absorb atmos-
tion of moisture. KBr powder must be pheric moisture.
dried in the oven, cooled and kept in a
desiccator.
3. Thin solid films: here a dilute solution of the compound in a low-boiling-
point solvent such as dichloromethane or ether is allowed to evaporate on
a NaCl plate producing a thin transparent film. This method gives excellent
results but is slightly limited by solubility factors.
When you are recording spectra of mulls, KBr disks and thin solid films air is
used as the reference and they are suitable for qualitative analysis only. The
procedure for the preparation of liquid and solid films and mulls is described
in Box 36.2 and that for KBr disks in Box 36.3.

Box 36.2 How to prepare liquid and solid films and mulls
A. Preparing a liquid film If the mull is too thick, add another drop of mulling
agent, or if it is too thin, add a little more solid. Only
1. Select a pair of clean NaCl plates from a desiccator,
experience will give you the correct consistency of
clean them by wiping with a soft tissue soaked in
the mull and the key to a good spectrum is a mull of
dichloromethane and place them on the bench on a
the correct fluidity.
piece of filter paper or tissue paper to prevent scratch-
ing by the bench surface. 3. Transfer the mull to the centre or along the diameter
of an NaCl plate, on a piece of filter paper or tissue
2. Using a glass rod or boiling stick, place a small drop
paper to prevent scratching by the bench surface,
of liquid in the centre of one of the plates. Do not use
using a small plastic spatula or a boiling stick.
a Pasteur pipette, which may scratch the surface of
the plate. 4. Carefully, holding it by the edge, place the other plate
on top and very gently press to ensure that the mull
3. Carefully, holding it by the edge, place the other plate
spreads as a thin film between the plates. If there is
on top and see if a thin film spreads between the
not enough liquid, carefully separate the plates by
plates, covering the centres. Do not press to force the
lifting at the edge and add another drop of mull. If
plates together. If there is not enough liquid, care-
there is too much liquid (poor spectrum), separate
fully separate the plates by lifting at the edge and
the plates and wipe the mull from one of them using
add another drop of liquid. If there is too much liquid,
a tissue.
separate the plates and wipe the liquid from one of
them using a soft tissue. D. Setting up the cell holder for liquid films and mulls
B. Preparing a thin solid film 1. Place the back-plate of the cell holder on the bench,
1. Dissolve the sample (about 5 mg) in a suitable low- position the rubber gasket, place the NaCl plates
boiling-point solvent (about 0.25 mL), such as DCM on the gasket and then put the second gasket on
or ether. top of the plates. These gaskets are essential to
prevent fracture of the plates when you tighten the
2. Place two drops of the solution onto the centre of a locking nuts.
NaCl plate and allow the solvent to evaporate. Use a
Pasteur pipette, but do not touch the surface of the plate. 2. Carefully place the cell holder top-plate on the top
gasket, drop the locking nuts into place and carefully
3. If the resulting thin film of solid does not cover the tighten each in rotation. These are safety nuts and if
centre of the plate, add a little more solution. you over-tighten them or if the back- and top-plates
4. Mount the single NaCl plate in the spectrometer and are not parallel, they will spring loose to prevent the
run the spectrum. Note that the NaCl plate can rest NaCl plates being crushed.
on the ‘V’-shaped wedge on the sample holder in the 3. Transfer the cell holder assembly to the spectro-
spectrometer. meter and make sure it is securely mounted in the
cell compartment.
C. Preparing a mull
4. Clean the plates in the fume cupboard by wiping
1. Grind a small sample of your compound (about 5 mg)
them with a tissue soaked in DCM, stand them on
using a small agate mortar and pestle for at least 2
filter paper or tissue paper to allow the solvent to
minutes. The powder should be as fine as possible.
evaporate and put them in the desiccator. Allow the
2. Add one drop of mulling agent (Nujol® or HCB) and DCM to evaporate from the tissue swab and dispose
continue grinding until a smooth paste is formed. of it in the chemical waste.
Attenuated total reflectance (ATR) sample holders

Handling NaCl plates and KBr disks –
NaCl plates are delicate and easily
The principle of this technique depends on the fact that when light passes from
damaged by scratching, dropping or
a dense medium to a less dense medium, and the angle of the incident light is at
squeezing. Hold them only by the edges
a critical angle, then the light is reflected back into the dense medium, having
and place them on filter paper or tis-
penetrated a short distance into the less dense medium (see Fig. 36.6). In effect,
sue when adding chemicals. KBr disks
the infrared spectrum of the less dense medium is recorded.
should be handled using tweezers.
The ATR cell is commonly made of infrared transparent zinc selenide
(ZnSe) and the major advantage of ATR cells is that spectra are now available

bevelled edge
plunger
pumping handle
pallets top die
base die
die holder ram

body
pressure gauge
O-ring seal pressure valve
anvil
Fig. 36.5 Equipment for preparation of a KBr disk.
Box 36.3 How to prepare a KBr disk
1. Take spectroscopic-grade KBr powder from the oven 9. Place the assembled disk kit in the hydraulic press
and allow it to cool in a desiccator. and tighten the top screw so that it touches the top of
the plunger.
2. Grind your compound (1–2 mg) in an agate mortar
for 2 minutes, then add the KBr (0.2 g) and continue 10. Connect the anvil to a source of vacuum, e.g. a rotary
grinding to a fine powder. Put the KBr powder back vacuum pump.
into the oven.
11. Close the hydraulic release valve on the side of the
3. Obtain a ‘disk kit’ and make sure that: press and gently pump the handle until the pressure
gauge reads between 8 and 10 tons and leave for
(a) it is complete – comprising a plunger, two dies 30 seconds.
(base and top), a die holder and an anvil, as
12. Open the hydraulic release valve gently and, when the
shown in Fig. 36.5;
pressure has fallen to zero, disconnect the vacuum
(b) the components are for the same device – they from the anvil.
are not interchangeable with another disk kit –
and should be numbered. 13. Loosen the top screw and remove the disk kit from
the press.
4. Press the die holder onto the anvil ensuring a proper fit.
14. Turn the disk kit upside down and carefully pull off
5. Lower the base die, numbered-side down, into the the anvil. Make sure that the plunger does not slide
die holder and make sure it slides into a depth of out by supporting it in the palm of your hand.
about 50 mm.
15. Gently push the plunger and the base die will emerge
6. Pour the compound/KBr powder mixture, about one- from the die holder. Take off the base die leaving the
third to one-half of the amount prepared, into the die KBr disk exposed.
holder and tap gently to produce an even layer on the
16. Carefully slide the KBr disk into the special disk
base die.
holder using a microspatula.
7. Lower the top die, numbered-side up, on top of the
17. Run the IR spectrum immediately, because the disk
KBr mixture and make sure it slides down onto the
will begin to cloud over as it absorbs atmospheric
powder.
moisture.
8. Slide the plunger, with the bevelled edge at the top, 18. Clean the disk kit components with a tissue and
into the die holder ensuring that it is touching the check that all parts are present.
top die and press down gently so that the dies slide
to the bottom, ensuring that you do not then push 19. If the dies or the plunger stick in the die holder, tell
off the anvil. your instructor.

sample
ZnSe crystal
from to dectector
IR source
Fig. 36.6 Schematic of an ATR cell.
Sample from a wide variety of sample types without the need for sample preparation –
Pressure arm
Metal mounting plate: the sample is simply laid on top of the ATR cell. Paper (with print), fabrics,
316 S/S or Hastalloy Diamond polymer films, powders, gels and even aqueous solutions can produce high
quality infrared spectra.
A recent development which simplifies the acquisition of IR spectra with
minimum sample preparation is the ATR ‘diamond anvil’ cell. This comprises
a ZnSe focussing element with a small circular diamond crystal (∼1.5 mm
diameter) on top mounted in a stainless steel top plate (Fig. 36.7). The sample
under examination, liquid, solid, solution, fiber, polymer film, etc. is simply
ZnSe (or KRS-5)
focusing element placed on the diamond and the spectrum obtained. In order to maintain good
contact with the diamond, solids are pressed into contact using the pressure
Fig. 36.7 Schematic diagram of an ATR arm. In most cases, sample handling is facilitated without result of the durabil-
‘diamond anvil’. ity of diamond and stainless steel, clean-up of the ‘diamond anvil’ is an easy
‘wipe clean’ with no possibility of damaging the cell window. The only minor
disadvantage of the ‘diamond anvil’ cell is the slight reduction in intensity of
the IR absorption peaks between 2,500 cm -1 and 1650 cm -1 resulting from
absorption of IR radiation by the diamond.
Interpretation of IR spectra
To identify compounds from their IR spectrum you should know at which
frequencies the stretching and bending vibrations occur. A detailed analysis
can be achieved using the correlation tables found in specialist textbooks. For
interpretation, the spectrum is divided into three regions.
Region 1 (400092000 cm -1): this region contains the high frequency vibra-
tions such as C ¬ H, N ¬ H and O ¬ H stretching, together with
C “ C and C ‚ N stretching vibrations.
Region 2 (200091500 cm -1): this is known as the ‘functional group region’ and
includes the stretching frequencies for C “ C, C “ O, C “ N, N “ O
and N ¬ H bending vibrations.
Region 3 (15009650 cm -1): this region contains stretching bands for
C ¬ O, C ¬ N, C ¬ Hal and the C ¬ H bending vibrations. It is
known as the ‘fingerprint region’ because it also contains complex
low-energy vibrations resulting from the overall molecular structure
and these are unique to each different molecule. Fig. 36.8 shows the
spectra of 1-propanol and 1-butanol, both of which show almost identi-
cal peaks for the O ¬ H, C ¬ H and C ¬ O stretching frequencies and
the C ¬ H bending frequencies, but the spectra are different in the num-
ber and intensity of the peaks between 1500 and 650 cm -1, resulting
from the presence of the additional CH2 in 1-butanol. Conversely, these
highly specific bands in the ‘fingerprint’ region are useful for identifica-
tion of molecules by comparison with authentic spectra via a database.
A simple correlation chart indicating the three regions of the spectrum and their
associated bond vibrations is shown in Fig. 36.9. You can obtain most diag-
nostic information from spectral regions 1 and 2, since these are the simplest

%T
100.00
Y
Z
finger print
region
0.00
4000 3500 3000 2500 2000 1500 1000 600 cm-1
Fig. 36.8 IR spectra of 1-propanol (Y) and 1-butanol (Z).
regions containing the peaks related to specific functional groups, while region
3 is normally used for confirmation of your findings. Another important aspect
of the IR spectrum is the relative intensities of the commonly found peaks and
you should become familiar with peak sizes. A chart indicating the positions,
general shapes and relative intensities of commonly found peaks is shown in
Fig. 36.10. When you are attempting to interpret an IR spectrum you should
use the approach described in Box 36.4.
– – –
KC–H =C–H –C–H
– CKC C=C –C–H
– –C–H
– =C–H =C–H
H
–C=–O
O–H C=O C–O C–O

COOH
ketone ether
aldehyde 1° alcohol
ester 2° alcohol
acid 3° alcohol
acid chloride phenol

amide
N–H
N–H
NH2
CKN C=N
NO2 NO2
–
–C–F
– –
–C–Cl
–
O
= =
P=OH
O
O
=
S=OH
=
4000 3500 3000 2500 2000 1500 1000 600 cm-1
Fig. 36.9 Simplified correlation chart of functional group absorptions.

%T
CH3|CH2 bend
100.00
Ar–H/=C–H
monosubstituted
CH2|CH3
CKN
C–O
CKC
C=C
aromatic
C=O
NH2
COOH
OH
NO2
0.00
4000 3500 3000 2500 2000 1500 1000 600 cm-1
Fig. 36.10 Idealised intensities of some IR bands of common functional groups.
Box 36.4 How to interpret an IR spectrum
1. Note the conditions under which the spectrum was unsaturation, while just below (2980 ¬ 2800 cm -1)
obtained, which should be written on the spectrum you will find the Csp3 ¬ H stretching frequencies for
as ‘phase’. If it is a solution or a mull, you will need CH3, CH2 and CH in saturated systems. Other bands
to identify and ‘subtract’ the spectrum of the mulling for O ¬ H, N ¬ H, C ‚ C and C ‚ N are obvious.
agent or solvent.
7. Examine region 2 (2000−1500 cm−1). Here you will
2. Consider carefully the reaction you have carried out. find C “ O stretch, usually the most intense band in
You should know, from the correlation table, the func- the spectrum; C “ C and C “ N stretches, less intense
tional groups and peaks in the starting materials and and sharper; N “ O stretch (from NO2) intense and
those expected in the product. sharp and with a twin band in region 3; N ¬ H bend-
3. Remember that the absence of peaks may be as use- ing vibrations – do not confuse with C “ O.
ful in interpretation as the presence of peaks. 8. Examine region 3 (1500−650 cm−1). The large bands
4. Do not attempt to identify all the peaks, just those here are C ¬ O, C ¬ N, C ¬ Cl, S “ O, P “ O, N “ O
which are relevant to your interpretation. Go for the (twin from region 2) stretches and C ¬ H ‘breath-
large peaks first. ing’ bands (9009700 cm-1), which indicate the num-
ber and position of substituents on a benzene ring.
5. Many sharp peaks of medium to strong intensity Medium-intensity peaks of importance include the
throughout the spectrum generally indicate an aro- CH3 and CH2 bands at 1460 cm -1 and 1370 cm -1 from
matic compound. the carbon skeleton which are also found in Nujol®.
6. Examine region 1 (4000−2000 cm−1). It is useful to 9. Tabulate your results and make the appropriate
draw a line on the chart at 3000 cm -1: just above deductions, after consulting the detailed correlation
the line (300093100 cm-1) you will find the stretch- table. Remember to correlate the spectroscopic data
ing frequencies for Csp ¬ H and Csp2 ¬ H indicating with the chemical data.

If you are studying complexes formed from metals and organic ligands,
the metal–ligand stretching vibration will occur below 600 cm -1 and special
IR spectrometers are used to observe this region. However, changes in the IR
spectrum of the organic ligand on complexation can be detected in the normal
40009650 cm -1 range.
Raman spectroscopy
The development of high powered lasers has resulted in the development
and application of Raman spectroscopy. Fig. 36.11 shows the energy levels
involved in the production of Raman spectra. The upper energy levels to which
a moleculte is excited by the incident light are referred to as the ‘virtual lev-
els’. The excitation in this case is not quantised, however, the method provides
information on the quantised vibrational levels of the molecule’s ground state
(as the molecule must return to one of those levels when the scattering event
is complete). Most of the photons scattered by the Raman mechanism have a
higher wavelength (i.e. lower energy) than the incident light – these are called
the ‘Stokes’ lines. In addition, it is also possible for Raman scattering to take
place at shorter wavelengths (i.e. higher energy) – these are called the ‘Anti-
Stokes’ lines. Anti-Stokes lines are normally weaker in intensity and so are not
usually used in analytical work.
Raman spectroscopy is essentially a scattering method. Most scattering
takes place via the Rayleigh mechanism – this is where the photon-molecule
collisins are elastic (i.e. do not involve any exchange of energy). As a result the
scattered photons have the same wavelength (and energy) as the incident pho-
tons (see Fig. 36.11). Under normal conditions however, a small proporption
of photons are scattered inelastically; in this case some energy is exchanged
between a photon and the molecule it collides with. The quanta of energy
involved correspond to the vibrational energy levels of the molecules involved.
Often the molecule will be in its ground vibrational state, so will accept energy
from the photon (rather than vice versa). The Raman scattered light thus has a
Virtual
energy states
Rayleigh
scattering
Stokes
raman
scattering
Anti-stokes
Excitation raman
energy scattering
4
3
Vibrational
2 energy
states
1
IR
absorbance
0
Fig. 36.11 Energy level diagram showing the states involved in the Raman signal.

Water molecule higher wavelength (i.e. lower energy) than the incident light; this principle is
illustrated in Fig. 36.12 using the water molecule as an example.
Scattered radiation
Principle of Raman spectroscopy

Incident radiation In infrared spectroscopy, absorption only occurs when a vibration causes a
change in the dipole moment. In Raman spectroscopy, the principle is differ-
Fig. 36.12 Illustration of the scattered radi- ent even though it is the same bonds and vibrations that are involved. When
ation from a water molecule. the electric field associated with the incident radiation interacts with a bond it
causes a momentary distortion in the associated electron could and a short-lived
induced dipole moment, which disappears when the radiation is re-emitted.
Symmetric C-O
stretch The magnitude of this induced dipole moment is proportional to the polaris-
ability of the bond (i.e. the extent to which the electron cloud is distorted by
the electric field). As bonds lengthen they become more polarisable. Raman
scattering only takes place when a molecular vibration is accompanied by a
change in polarisability.
Example: the symmetrical stretching mode of CO2 involves a polarisability
Bend
change in both C “ O bonds as they lengthen and contract (see Fig. 36.13). This
is therefore a Raman active vibration (at approximately 1480 cm -1). In contrast
in the asymmetrical stretching mode one C “ O bond lengthens as the other con-
tracts; therefore no change in polarisability takes place – so no Raman activity. In
addition, in the bending mode the CO2 molecule has no bond length changes –
so it is also Raman inactive. In conclusion, for a simple linear centrosymmetric
Asymmetric molecule (e.g. CO2), infrared active modes are Raman inactive (and vice versa).
C-O stretch
Fig. 36.13 The vibration modes of carbon Resonance Raman spectroscopy

dioxide. Normally, the Raman scattering effect is weak and hence the technique has
had limited applicability. The development of Resonance Raman spectroscopy
(RRS) has evolved due to the development of tunable lasers. In RRS it is pos-
sible to excite the sample at a wavelength at or near an electronic absorption
maximum of the analyte (in aqueous solution), using a tunable laser, thereby
increasing the sensitivity of the technique. A schematic diagram of a typi-
cal Raman spectrometer is shown in Fig. 36.14. A further development that
has enhanced Raman intensities is Surface Enhanced Raman spectroscopy
(SERS). In contrast to RRS the sample is not investigated in an aqueous solu-
tion but is instead adsorbed on to a surface of a colloidal preparation of silver
or gold. The combination of RRS and SERS has also led to the development of
Surface Enhanced Resonance Raman spectroscopy (SERRS) which provides
the enhanced sensitivity of both techniques.
Sampling probe CCD detector

Emission fibre
Excitation fibre
Diode laser Narrow
bandpass
filter Monochromator
Sample
Rayleigh rejection filter
Fig. 36.14 Schematic diagram of a Raman spectrometer.

The main advantages of Raman spectroscopy are as follows:

●● Non-destructive technique
●● No sample preparation
●● Ability to collect information at low wavenumber (i.e. 100 cm -1)
●● Fast collection of data
●● High spectral resolution is possible (typically 1 cm -1)
●● High spatial information (6 1 mm)
●● Line width of spectral features generally sharp offering good chemical dis-
tinguishing power
●● Water is a weak Raman scatterer and does not mask the spectrum, enabling
the analysis of aqueous solutions.

Anderson, R.J., Bendell, D.J. and Groundwater, P.W. Shriner, R.L., Hermann, C.K.F., Morrill, T.C., Curtin, D.Y.
(2004) Organic Spectroscopic Analysis. Royal Society and Fuson, R.C. (2003) Systematic Identification of
of Chemistry. Cambridge. Organic Compounds, 8th edn. John Wiley & Sons Ltd,
CD-ROM Abrams, C.B. (1992) IR Tutor. Perkin Elmer Ltd, Chichester.
Beaconsfield, UK. Silverstein, R.M., Webster, F.X., Kiemle, D.J. and Bryce,
Crews, J., Rodriguez, J. and Jaspars, M. (2010) Organic D.L. (2015) Spectroscopic Identification of Organic
Structure Analysis. 2nd edn. Oxford University Press, Chemicals, 7th edn. John Wiley & Sons Ltd, Chichester.
New York. Smith, B.C. (1998) Infrared Spectral Interpretation: A Sys-
Field, L.D., Sternhell, S. and Kalman, J.R. (2013) Organic tematic Approach. CRC Press, Boca Raton.
Structures from Spectra, 5th edn. John Wiley & Sons Stuart, B.H. (2004) Infrared Spectroscopy: Fundamentals
Ltd, Chichester. and Applications. John Wiley & Sons Ltd, Chichester.
Griffiths, P. and De Haseth, J.A. (2007) Fourier Transform Williams, D. and Fleming, I. (2008) Spectroscopic Methods
Infrared Spectroscopy, 2nd edn. John Wiley & Sons Ltd, in Organic Chemistry, 6th edn. McGraw-Hill, Maiden-
Chichester. head, UK.
Pavia, D.L., Lampman, G.M., Kriz, G.S. and Vyvyan, J.A. Wolstenholme-Hogg, P. Chemtube UK. Available: https//
(2015) Introduction to Spectroscopy, 5th edn. Brooks www.youtube/com/user/chemtubeuk
Cole, Pacific Grove. Last accessed 14/02/2016.

Study exercise
36.1 The following infrared spectra A S D are all run (iv) ethyl ethanoate. Deduce the structural infor-
as liquid films and they represent the colouress mation from each of the spectra A S D and
liquids (i) 1-octanol, (ii) heptane, (iii) benzonitile, hence identify the compounds A S D.
%T %T
4000 3000 2000 1500 1000 cm-1 4000 3000 2000 1500 1000 cm-1
Spectrum A Spectrum B
%T %T
4000 3000 2000 1500 1000 cm-1 4000 3000 2000 1500 1000 cm-1
Spectrum C Spectrum D

37 Nuclear magnetic resonance
spectroscopy
Electromagnetic radiation (typically at radio frequencies of 60–600 MHz) is

Nuclear spin quantum numbers and
used to identify compounds in a process known as nuclear magnetic resonance
NMR – other common nuclei with
(NMR) spectroscopy. This is possible because of differences in the magnetic
non-zero quantum numbers are 14N, 2D
states of atomic nuclei, involving very small transitions in energy levels. The
(I = 1) and 11B and 35Cl (I = 32). Their
atomic nuclei of the isotopes of many elements possess a magnetic moment.
NMR spectra are not used on a routine
When these magnetic moments interact with a uniform external magnetic
basis.
field, they behave like tiny compass needles and align themselves in a direc-
tion ‘with’ or ‘against’ the field. The two orientations, characteristic of nuclei
with a nuclear spin quantum number I = 12, have two different energies: the
orientation aligned ‘with’ the field has a lower energy than that aligned ‘against’
the field (Fig. 37.1).
Typical magnetic nuclei of general use to chemists and biochemists are
N 1
H, 13C, 19F and 31P, all of which have nuclear spin quantum numbers I = 12.
pole of magnet The energy difference between the two levels (∆E) corresponds to a precise
electromagnetic frequency (n), according to similar quantum principles for the
applied excitation of electrons (p. 217). When a sample containing an isotope with a
field
(B0) magnetic nucleus is placed in a magnetic field and exposed to an appropriate
sample containing radio frequency, transitions between the energy levels of magnetic nuclei will
magnetic nuclei
occur when the energy gap and applied frequency are in resonance (i.e. when
S they are matched exactly in energy). Differences in energy levels, and hence
resonance frequencies (n0), depend upon the magnitude of the applied magnetic
(a) field (B0) and the magnetogyric ratio (g), according to the equation:
n0 = gB0/2p[37.1]
applied For a given value of the applied field (B0), nuclei of different elements have
field ¢E = hv different values of the magnetogyric ratio (g) and will give rise to resonance at
(B0)
various radio frequencies. The principal components of an NMR spectrometer
are shown in Fig. 37.2.
For magnetic nuclei in a given molecule, an NMR spectrum is generated
(b)
because, in the presence of the applied field, different nuclei of the same atoms
Fig. 37.1 Effect of an applied magnetic experience small, different, local magnetic fields depending on the arrangement
field, B0, on magnetic nuclei. (a) Nuclei in of electrons, i.e. in the chemical bonds, in their vicinity. The effective field at
magnetic field have one of two orienta- the nucleus can be expressed as:
tions – either with the field or against the
field (in the absence of an applied field, the B = B0(1@s)[37.2]
nuclei would have random orientation).
(b) Energy diagram for magnetic nuclei in where s (the shielding constant) expresses the contribution of the small sec-
applied magnetic field. ondary field generated by the nearby electrons. The magnitude of s depends
pole of magnet
radio frequency
field N transmitter
controller
radio frequency computer
Example For an external m agnetic receiver
display
field of 2.5T (tesla), ∆E for 1H is
S
6.6 * 10-26 J and since ∆E = hn, probe
the corresponding frequency (n) is (contains
100 MHz; for 13C in the same field, ∆E sample)
is 1.7 * 10-26 J, and n is 25 MHz.
Fig. 37.2 Components of an NMR spectrometer.

Nuclear magnetic resonance spectroscopy
on the electronic environment of a nucleus, so nuclei of the same isotope give

rise to small different resonance frequencies according to the equation:
n0 = gB0(1 - s)/2p[37.3]
Key Point The variation of resonance frequencies with sur-

rounding electron density is crucial to the usefulness of the NMR
technique. If it did not occur, all nuclei of a single isotope would
come into resonance at the same combination of magnetic field
and radio frequency and only one peak would be observed in
the spectrum.
Chemical shift
Measuring chemical shifts – ppm is
not a concentration term in NMR but The separation of resonance frequencies resulting from the different electronic
is used to reflect the small frequency environments of the nucleus of the isotope is called the chemical shift. It is
changes that occur relative to the ref- expressed in dimensionless terms, as parts per million (ppm), against an inter-
erence standard, measured in propor- nal standard, usually tetramethylsilane (TMS). By convention, the chemical
tional terms. shift is positive if the sample nucleus is less shielded (lower electron density
in the surrounding bonds) than the nucleus in the reference and negative if it is
more shielded (greater electron density in the surrounding bonds). The chem-
ical shift scale (d) for a nucleus is defined as:
d = [(nsample - nreference) * 106]/(nreference)[37.4]
This means that the chemical shift of a specific nucleus in a molecule is at the
same d value, no matter what the operating frequency of the NMR spectrometer.
An NMR spectrum is a plot of chemical shift (d) as the x-axis against
absorption of energy (resonance) as the y-axis. On the right-hand side of the
spectrum at d = 0 ppm there may be a small peak, which is the reference
(TMS). A typical 1H -NMR spectrum is shown in Fig. 37.3.
NMR spectrometers
These can operate at different radio frequencies and magnetic fields and are
usually referred to in terms of radio frequency, e.g. 60 MHz, 270 MHz and
500 MHz spectrometers. Spectrometers operating above 100 MHz require
b a
peak area
c integration
O
a c b
CH3OCH2CCH3 TMS
f (ppm)
5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
Fig. 37.3 1H -NMR spectrum of 1-methoxypropanone.

expensive superconducting magnets to generate the high magnetic fields. In

Using deuterated solvents – these are
routine laboratory work 60 MHz and 90 MHz instruments are common but 270
expensive and should not be wasted.
MHz machines are becoming more affordable. Increasing the operating fre-
CDCl3 is 100 times more expensive than
quency of the spectrometer effectively increases the resolution of the chemical
spectroscopic-grade CHCl3 and the oth-
shifts of the nuclei under examination. For example, the difference in frequency
ers are at least 10–15 times more expen-
between 0 and 1d is 60 Hz in a 60 MHz spectrometer but 270 Hz for a 270
sive than CDCl3.
MHz instrument.
Spectrometers can be divided into two types:
1. Continuous-wave (CW) spectrometers, which use a permanent magnet or

Using CDCl3 – when using this solvent
an electromagnet, usually operating at 60 or 90 MHz. In practice the radio
additional peaks can appear in the spec-
frequency is held constant and small electromagnets on the faces of the
trum. 1H spectra – a sharp single peak
main magnet (sweep coils) vary the magnetic field over the chemical shift
at dH = 7.26 ppm due to the presence of
range. The spectrometer sweeps through the spectroscopic region plotting
CHCl3 as an impurity (Fig. 37.4).
resonances (absorption peaks) on a chart recorder (cf. dispersion IR spec-
trometers). CW spectrometers are usually dedicated to observation of a
specific nucleus such as 1H.
13
C spectra – a triplet at dC = 77.41 ppm 2. Fourier transform (FT) spectrometers, using superconducting magnets
due to coupling between 13C and containing liquid nitrogen and liquid helium for cooling. Here the magnetic
D (I = 1) (Fig. 37.5). field is held constant and the sample is irradiated with a radio frequency
pulse containing all the radio frequencies over the chemical shift region of
the nucleus being examined, cf. FT–IR (p. 322). Computer control allows
rapid repeat scans to accumulate spectra, presenting the data as a stand-
ard CW-type spectrum via FT processing. Simple variation of the radio
frequencies permits observation of different nuclei (multinuclear NMR
spectrometers). Thus an FT–NMR spectrometer can be used for obtaining
CHCl3 in CDCl3 at fH = 7.26 ppm 1
H, 13C, 19F, 15N and 31P NMR spectra.
10.0 9.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0
Sample handling
Fig. 37.4 1H@NMR spectrum. The majority of NMR spectra are obtained from samples in solution and there-
fore the solvent should preferably not contain atoms of the nuclei being observed
(except in the case of 13C9NMR). The most common solvents are those in which
the hydrogen atoms have been replaced by deuterium, which is not observed
under the conditions under which the spectrum is obtained. CDCl3 (deutero-
CDCl3 at fC = 77.41 ppm chloroform, chloroform-d) is often the solvent of choice, but others such as
dimethylsulphoxide@d 6, [(CD3)2SO], propanone@d 6 [(CD3)2CO], methanol@d 4
(CD3 OD) and deuterium oxide (D2O) are in common use.
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
As it is unlikely that you will be allowed ‘hands-on’ use of an NMR
Fig. 37.5 13C@NMR spectrum (broad band spectrometer, the best approach you can take to obtain a good spectrum
decoupled). is to ensure good sample preparation. The quality of an NMR spectrum is
degraded by:
●● inappropriate solvent;
●● inappropriate concentration of solute;
●● inappropriate solvent volume;
●● solid particles in the solution;
●● water in the sample (inefficient drying);
●● paramagnetic compounds.
Sample preparation for NMR spectroscopy is described in Box 37.1.

Box 37.1 How to prepare a sample for NMR spectroscopy
1. Make sure that your compound is free from water and cotton wool with glasswool and do not use alumina.
solvent (p. 95). You must wear gloves when handling glasswool.
2. Test the solubility of your compound in cold CH2Cl2. 5. Put the filter into a suitable clean, dry NMR tube and,
If it is soluble you can use CDCl3 as the solvent for using a clean, dry Pasteur pipette, filter the solution
the NMR experiment. If it is insoluble, consult your into the NMR tube.
instructor for the availability of other deuterated
solvents. 6. Fill the NMR tube to the appropriate level: between
30 and 50 mm in height is sufficient.
3. Dissolve your compound CDCl3 (about 2 mL) in a
clean, dry sample tube. Use about 10 mg of sample 7. Cap the NMR tube with the correct-size tube cap,
for CW–NMR or 5 mg of sample for FT–NMR. Check to making sure that it is correctly fitted to prevent oscil-
see if the solvent contains TMS; if it does not, consult lation when the tube is spinning in the spectrometer.
your instructor. Make sure that the cap is fitted correctly so that it will
not fall off when the tube is in the spectrometer.
4. Make a simple filter in a new Pasteur pipette to
remove insoluble material and water (Fig. 37.6). Check 8. Wipe the outside of the tube with a clean, dry tissue
that your compound does not react with cotton wool to make sure that the spectrometer will not be con-
and neutral alumina (alcohols and acids are strongly taminated. Cleaning the spectrometer probe is a very
adsorbed on neutral alumina). If it does, replace the difficult task.
1
Interpreting NMR spectra
H9NMR spectra – most of the spec-
tra shown in this chapter do not
As a matter of routine in your laboratory work you will be required to inter-
extend over the normal spectral range
pret 1H9NMR spectra (also known as proton spectra). 13C9NMR spectra are
d = 0910 ppm. They are expanded to
becoming more common, while 19F and 31P spectra may be obtained in spe-
show the details of coupling patterns.
cialised experiments. Therefore you should concentrate on the interpretation
of 1H and 13C spectra in the first instance.
1
H9NMR spectra
These normally cover the range between d = 0 and 10 ppm but the range is
increased to d = 15 ppm when acidic protons are present in the molecule. The
1
sample H@NMR spectrum of a molecule gives three key pieces of information about
the structure of a molecule:
1. Chemical shift (d): the peak positions indicate the chemical (magnetic)
environment of the protons, i.e. different protons in the molecule have
different chemical shifts.
pipette 2. Integration: the relative size of peak area indicates how many protons have
neutral Al2O3 the d value shown.
cotton wool
3. Coupling: the fine structure on each peak (coupling) indicates the number
of protons on adjacent atoms.
NMR tube These three features make 1H@NMR a powerful tool in structure determination
and there are two extreme approaches to it:
1. Prediction of the spectrum of the expected compound from theoretical
knowledge and then comparison with the spectrum obtained. You should
recognise ‘patterns’ (e.g. triplet and quartet for an ethyl group; a singlet
of peak area six for two identical methyl groups), which were present in
Fig. 37.6 Filtration of solutions for NMR. the starting materials, but the dH values may have changed in the ‘new’

molecule. There are computer programs, such as g-NMR®, which will

Proton chemical shifts – only hydro-
simulate the NMR spectrum from a structural formula.
gen atoms bonded to carbon will be
considered in this simplified treatment. 2. Interpretation of the spectrum from correlation tables, but this is very dif-
ficult for the inexperienced.
In practice a combination of the two approaches is used with cross-referencing
Interpreting NMR spectra: changes of
and checking the proposed structure with tabulated dH values and reference
d – the terms used to indicate the move-
spectra until a satisfactory answer is found.
ment of a particular peak with change
in its chemical (magnetic) environ-
ment are: upfield – towards d = 0 ppm; Key Point Always make sure that your predicted structure is
d ownfield – towards d = 10 ppm; consistent with the spectrum.
shielded – increased electron density
near the proton; deshielded – decreased
electron density near the proton. Factors affecting chemical shift (dH)
The d values of protons can be predicted to a general approximation from
knowledge of the effects which produce variations in chemical shift.
1. The hybridisation of the carbon atom to which the hydrogen atom is
Table 37.1 Chemical shifts of methyl protons attached:
(a) sp3 hybridised carbon: peaks occur between d = 0.9 and 1.5 ppm in
Compound Chemical shift (ppm)
simple hydrocarbon systems. The peaks move downfield with change
(CH3)4Si 0.00 of structure from CH3 to CH2 to CH.
CH3R 0.90 (b) sp hybridised carbon: peaks occur at about d = 1.593.5 ppm in
CH3I 2.16 alkynes.
CH3Br 2.65 (c) sp2 hybridised carbon: in alkenes the resonances occur around
CH3CI 3.10 d = 498 ppm and the C9H peaks of aromatic rings are found between
CH3OR 3.30 d = 6 and 9 ppm. The large downfield shifts of these Csp2 9H nuclei
result from deshielding of the protons by fields set up by circulation
CH3F 4.26
of the p@electrons in the magnetic field. The proton of the aldehyde
group (CHO) is particularly deshielded by this effect and is found at
d = 9910 ppm.
2. Electron attraction or electron release by substituent atoms attached
to the carbon atom. Electron attracting atoms, such as N, O, Hal attached to
the carbon, attract electron density from the C9H bonds and thus deshield
the proton. This results in movement of the chemical shift to higher d val-
ues (Table 37.1). Conversely, electron-releasing groups produce additional
shielding of the C9H bonds resulting in upfield shifts of d values.
3. All the protons in benzene are identical and occur at d = 7.27 ppm. In
substituted aromatic compounds, the overall electron-attracting or releas-
ing effect of the substituent(s) alters the d values of the remaining ring
protons making them non-equivalent. The ortho protons are affected most.
4. For protons attached to atoms other than carbon: the chemical shifts of
protons attached to oxygen increase with increasing acidity of the O9H
aromatic OH
group; thus d = 196 ppm for alcohols, 4–12 ppm for phenols and 10–14
COOH CHO alkene Csp–H Csp3–H ppm for carboxylic acids. Hydrogens bound to nitrogen (1° and 2° amines)
are found at d = 398 ppm. The approximate chemical shift regions are
13 12 11 10 9 8 7 6 5 4 3 2 1 0 shown in Table 37.2 and Fig. 37.7.
Fig. 37.7 Approximate chemical shift posi- A range of chemical shift correlation tables is provided in Tables 37.2, 37.3,
tions in the 1H9NMR spectrum. 37.4, 37.5 and 37.6.

Table 37.2 Proton chemical shifts of common alkyl derivatives RX (dH ppm)
R Methyl Ethyl n-Propyl iso-Propyl t-Butyl
X CH3 CH2 CH3 a@CH2 b@CH2 CH3 CH CH3 CH3

H 0.23 0.86 0.86 0.91 1.33 0.91 1.33 0.91 0.89
¬ CH “ CH2 1.71 2.00 1.00 1.73 1.02
¬ C ‚ CH 1.80 2.16 1.15 2.10 1.50 0.97 2.59 1.15 1.22
¬ Ph 2.35 2.63 1.21 2.59 1.65 0.95 2.89 1.25 1.32
¬F 4.27 4.36 1.24 4.30 1.68 0.97
¬ Cl 3.06 3.47 1.33 3.47 1.81 1.06 4.14 1.55 1.60
¬ Br 2.69 3.37 1.66 3.35 1.89 1.06 4.21 1.73 1.76
¬I 2.16 3.16 1.88 3.16 1.88 1.03 4.24 1.89 1.95
¬ OH 3.39 3.59 1.18 3.49 1.53 0.93 3.94 1.16 1.22
¬O¬ 3.24 3.37 1.15 3.27 1.55 0.93 3.55 1.08 1.24
¬ OPh 3.73 3.98 1.38 3.86 1.70 1.05 4.51 1.31
¬ OCOCH3 3.67 4.05 1.21 3.98 1.56 0.97 4.94 1.22 1.45
¬ OCOPh 3.88 4.37 1.38 4.25 1.76 1.07 5.22 1.37 1.58
p-Tosyl 3.70 3.87 1.13 3.94 1.60 0.95 4.70 1.25
¬ CHO 2.18 2.46 1.13 2.35 1.65 0.98 2.39 1.13 1.07
¬ COCH3 2.09 2.47 1.05 2.32 1.56 0.93 2.54 1.08 1.12
¬ COPh 2.55 2.92 1.18 2.86 1.72 1.02 3.58 1.22
¬ COOH 2.08 2.36 1.16 2.31 1.68 1.00 2.56 1.21 1.23
¬ COOCH3 2.01 2.28 1.12 2.22 1.65 0.98 2.48 1.15 1.16
¬ CONH2 2.02 2.23 1.13 2.19 1.68 0.99 2.44 1.18 1.22
¬ NH2 2.47 2.74 1.10 2.61 1.43 0.93 3.07 1.03 1.15
¬ NHCOCH3 2.71 3.21 1.12 3.18 1.55 0.96 4.01 1.13
¬SH 2.00 2.44 1.13 2.46 1.57 1.02 3.16 1.34 1.43
¬ S¬ 2.09 2.49 1.25 2.43 1.59 0.98 2.93 1.25
¬C‚N 1.98 2.35 1.31 2.29 1.71 1.11 2.67 1.35 1.37
¬ NO2 4.29 4.37 1.58 4.28 2.01 1.03 4.44 1.53
Table 37.3 1H9NMR correlation table for methylene (CH2) groups
X or Y s (ppm) X or Y s (ppm)
¬H 0.34 ¬ OC( “ O)R 3.01

¬ CH3 0.68 ¬ OC( “ O)Ph 3.27
¬C“C 1.32 ¬ C( “ O)R 1.50
¬C‚C 1.44 ¬ C( “ O)Ph 1.90
¬ Ph 1.83 ¬ C( “ O)OR 1.46
¬ CF3 1.14 ¬ C( “ O) NR2 , ¬ C( “ O) NH2 1.47
¬F 3.30 ¬C‚N 1.59
¬ Cl 2.53 ¬ NR2 , ¬ NH2 1.57
¬ Br 2.33 ¬ NHPh 2.04
¬I 2.19 ¬ NHC( “ O)R 2.27
¬ OH 2.56 ¬ NO2 3.36
¬ OR 2.36 ¬ SH, ¬ SR 1.64
¬ OPh 2.94 ¬ OSO2 R 3.13
This table gives an estimate of the chemical shift of a CH2 group depending on the two groups X and Y attached to it
X ¬ CH2 ¬ Y
The estimated chemical shift d is: d = 0.23 + sX + sY

Table 37.4 1H9NMR correlation table for methine (CH) groups
X, Y, or Z s (ppm) X, Y, or Z s (ppm)
¬ alkyl 0 ¬ OC( “ O)R 2.07

¬C“C 0.46 ¬ C( “ O)R 0.47
¬C‚C 0.79 ¬ C( “ O)Ph 1.22
¬ Ph 0.99 ¬ C( “ O)OR 0.47
¬F 1.59 ¬ C( “ O)NR2 , ¬ C( “ O)NH2 0.60
¬ Cl 1.56 ¬C‚N 0.66
¬ Br 1.53 ¬ NR2 , ¬ NH2 0.64
¬ OH 1.14 ¬ NHC( “ O)R 1.80
¬ OR 1.14 ¬ NO2 1.84
¬ OPh 1.79
This table gives an estimate of the chemical shift of a CH group depending on the three groups X, Y, and Z attached to it:
X CH Y
The estimated chemical shift d is: d = 2.50 + sX + sY + sZ
Table 37.5 1H9NMR correlation table for alkenes
s (ppm)
Substituent gem cis trans
¬H 0 0 0
¬ Alkyl (linear) 0.44 - 0.26 - 0.29
¬ Alkyl (ring) 0.71 - 0.33 - 0.30
¬ CH2 OR 0.67 - 0.02 - 0.07
¬ CH2 I 0.67 - 0.02 - 0.07
¬ CH2 Cl 0.72 0.12 0.07
¬ CH2 Br 0.72 0.12 0.07

¬ CH2 Ar 1.05 - 0.29 - 0.32
¬ CH2 NR2 0.66 - 0.05 - 0.23
¬C‚C 0.50 0.35 0.10
¬C“C 0.98 - 0.04 - 0.21
¬C“O 1.10 1.13 0.81
¬ CO2 H 1.00 1.35 0.74
¬ CO2 R 0.84 1.15 0.56
¬ CHO 1.03 0.97 1.21
¬ C( “ O)NR2 1.37 0.93 0.35
¬ OR (R “ alkyl) 1.18 - 1.06 - 1.28
¬ OC( “ O)R 2.09 - 0.40 - 0.67
¬ Ph 1.35 0.37 - 0.10
¬ Cl 1.00 0.19 0.03
¬ Br 1.04 0.40 0.55
¬ NR2 (R “ alkyl) 0.69 - 1.19 - 1.31
This method allows the estimation of the chemical shift of a proton con-
nected to a carbon–carbon double bond:
cisR H
transR Rgem
d = 5.25 + scis + strans + sgem
d = 5.25 + scis + strans + sgem

Table 37.6 1H9NMR correlation table for substituted benzenes

Integration of coupled peaks – the
area under the singlet, doublet, triplet,
s (ppm)
quartet, etc., is still that of the type of
hydrogen being considered. For exam- Substituent ortho meta para
ple, if the peak for the three protons of
¬H 0 0 0
a methyl group is split into a triplet by ¬ Me - 0.20 - 0.12 - 0.22
an adjacent methylene group, the area ¬ Et - 0.14 - 0.06 - 0.17
of the triplet is three. ¬ CH2 OH - 0.07 - 0.07 - 0.07
¬ CH2 NH2 - 0.07 - 0.07 - 0.07
¬ CH2 Cl 0 0 0
¬ CF3 0.32 0.14 0.20
¬ CCl3 0.64 0.13 0.10
¬C“C 0.06 - 0.03 - 0.10
¬ Ph 0.37 0.20 0.10
¬ CHO 0.56 0.22 0.29
¬ COR 0.62 0.14 0.21
¬ C( “ O)N 0.61 0.10 0.17
¬ CO2 H 0.85 0.18 0.27
¬ CO2 R 0.71 0.10 0.21
¬C‚C 0.15 0.02 - 0.01
¬C‚N 0.36 0.18 0.28
¬ NH2 - 0.75 - 0.25 - 0.65
¬ NR2 (R “ alkyl) - 0.66 - 0.18 - 0.67
¬ NHC( “ O)R 0.12 - 0.07 - 0.28
¬ NO2 0.95 0.26 0.38
¬ OH - 0.56 - 0.12 - 0.45
¬ OR (alkyl) - 0.48 - 0.09 - 0.44
¬ OC( “ O)R - 0.25 0.03 - 0.13
¬F - 0.26 0 - 0.04
¬ Cl 0.03 - 0.02 - 0.09
¬ Br 0.18 - 0.08 - 0.04
¬I 0.39 - 0.21 0
This method allows the calculation of a chemical shift of a proton on a

phenyl ring depending on the nature of the substituents:
X
Hortho Hortho
Hmeta Hmeta
Hpara
d = 7.27 + sortho + smeta + spara
Integration of peak areas

The area of each peak gives the relative number of protons and is produced directly
on the spectrum (Fig. 37.8). On CW–NMR spectrometers the height of the peak area
integration line must be measured using a ruler, whereas on FT–NMR machines the
area is calculated and displayed as a number. You must remember that:
●● The areas are ratios, not absolute values, and you must find a peak attrib-
utable to a specific group to obtain a reference area, e.g. a single peak at

a b c
CH3CH2OCH3
c
a TMS
f (ppm)
4.0 3.0 2.0 1.0 0.0
Fig. 37.8 1H9NMR spectrum of methoxyethane.
d = 1.0 ppm is likely to be a CH3 group and thus the area displayed or
measured is equal to three protons.
●● You must ensure that you include integrations from all the fine-structure
(coupling) peaks in the peak area.
●● Do not expect the peak area integrations to be exact whole numbers, e.g.
an area of 2.8 is probably three protons (CH3), 5.1 is probably five protons
(e.g. a C6H5 group), but 1.5 is probably a CH3 and all the peak area integra-
tions must be doubled.
Coupling (spin–spin splitting)

Many 1H9NMR signals do not consist of a single line but are usually associated
with several lines (splitting patterns). Protons giving multiline signals are said
to be coupled. This coupling arises from the magnetic influence of protons on
one atom with those on an adjacent atom(s). Thus information about the nature
of adjacent protons can be determined and fed into the structural elucidation
problem. To a simple first approximation the following three general points are
useful in the interpretation of coupling patterns:
1. Aliphatic systems: if adjacent carbon atoms have different types of pro-
tons (a and b), then the protons will couple. If a proton is coupled to
n (n = 1, 2, 3, 4, 5, etc.) other protons on an adjacent carbon atom, the
number of lines observed is n + 1, as shown in the examples below.
CH3CH2OCH3 Protons a are coupled to two protons b: n = 2; therefore

a b c the peak for protons a is split into three lines (a triplet).
Protons b are coupled to three protons a: n = 3; there-
fore the peak for protons b is split into four lines (a
quartet).
Protons c have no adjacent protons and therefore are
not coupled and give a single line (singlet) (Fig. 37.8).
CH3CHBrCH2Br Protons a are coupled to one proton b: n = 1;
a b c therefore the peak for protons a is split into two lines
(doublet)

a b c
CH3CHBrCH2Br
a TMS
c
b
1 one line (singlet or s)

1 1 two lines (doublet or d)
f (ppm)
1 2 1 three lines (triplet or t) 4.0 3.0 2.0 1.0 0.0
1 3 3 1 four lines (quartet or q)
etc.
Fig. 37.9 1H9NMR spectrum of 1,2-dibromopropane.
Fig. 37.10 Intensities of coupled peaks from
Pascal’s triangle.
Note:
Protons b are coupled to three protons a and two protons c: n = 5; there-
fore the peak for protons b is split into six lines (sextet).
a b Protons c are coupled to one proton b: n = 1; therefore the peak for protons
H H c is split into two lines (doublet).
C C
Br C N Protons a and protons c are not adjacent and do not couple (Fig. 37.9).
The intensity of each peak in the resulting singlet, doublet, triplet, quartet,
etc. is calculated from Pascal’s triangle (Fig. 37.10).
The separation between the coupled lines is called the coupling con-
stant, J, and, for aliphatic protons CH, CH2 and CH3, it is usually ∼ 8 Hz.
f (ppm)
6.5 6.0 5.5
(a)
Key Point The (n + 1) rule only applies in systems where the
coupling constant (J) between the protons is the same. Fortu-
nately this is common in aliphatic systems.
b
Br H
C C
H C N 2. Alkene hydrogens: hydrogen atoms on double bonds have different cou-
a
pling constants depending upon the stereochemistry of the alkene. Alkene
hydrogens in the Z (cis) configuration have J = 5914 Hz, whereas those
in the E (trans) configuration have J = 11919 Hz (Figs 37.11(a) and (b)).
3. Aromatic hydrogens: coupling of hydrogens, which are non-adjacent, is
readily observed in aromatic compounds. Different protons ortho to each
f (ppm)
6.5 6.0 5.5
other couple with J = 7910 Hz, while those in a meta relationship have
(b) J = 293 Hz. Para coupling (J = 091 Hz) is not usually seen on the spec-
trum. The types of aromatic compound you are likely to meet most often are:
Fig. 37.11 1H9NMR spectra of: (a) (Z)-
3-bromopropanonitrile; (b) (E)-3- (a) Monosubstituted aromatic compounds, in which three basic patterns
bromopropano-nitrile. are found in the aromatic region of the spectrum. If the substituent

a
a
a a
b b CH3 b 0CH3
b c
b b d b
b c
TMS
c b,d
f (ppm) f (ppm)
8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 7.2 7.0 6.0 5.0 4.0
(a) (b)
a
a
b
b a
a NO2 b CH3
c b b
c a a b
b CH3 b
TMS
f (ppm) f (ppm)
8.5 8.0 7.5 7.0 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0
(c) (d)
a
c
a
c NO2 b Br c NO2
b
a c b
CH3O b H2N a
c b
f (ppm) f (ppm)
8.4 8.0 7.6 7.2 6.8 6.4 6.0 5.6 5.2 4.8 4.4 4.0 8.4 8.2 8.0 7.8 7.6 7.4 7.2 7.0 6.8 6.6 6.4
(e) (f)
Fig. 37.12 1H9NMR spectra of (a) methylbenzene; (b) methoxybenzene; (c) nitrobenzene; (d) 1,4-dimethylbenzene;
(e) 4-methoxynitrobenzene; (f) 4-amino-3-bromonitrobenzene (NH2 protons not shown).
exerts a weak electronic effect on the ring, the d values of the ring
protons are similar and the protons appear as a single peak of area
five (Fig. 37.12(a)). If the group is strongly electron releasing (OH,
NH2, OCH3, etc.), the protons appear as complex multiplets (ortho
and meta coupled), below d = 7.27 ppm of relative areas two to three
(Fig. 37.12(b)). If the group is electron attracting (e.g. NO2, COOH,
etc.), then the complex multiplets have d 7 7.27 ppm (Fig. 37.12(c));
(b) para disubstituted aromatic compounds, which are of two types. If the
substituents are the same, then all the ring protons are identical and a
singlet, of relative area four, is seen (Fig. 37.12(d)). If the substituents

are different, then the pairs of hydrogens ortho to each substituent are
different and ortho-couple to give what appears to be pair of doublets,
each of relative area two (Fig. 37.12(e)).
(c) Increasing numbers of substituents, which decrease the number of
aromatic hydrogens and the spectrum becomes simpler. Thus the
common 1,2,4-trisubstituted pattern (Fig. 37.12(f)) is recognised eas-
ily as an ortho-coupled doublet, a meta-coupled doublet and a doublet
of doublets (coupled ortho and meta).
The chemical shifts of aromatic protons can be calculated from
detailed correlation tables (Table 37.6).
4. Where the dH values for coupling protons are quite close, the resulting multi-
plets (doublet, triplet, quartet etc.) are not symmetrical but due to secondary
interactions they ‘lean’ towards each other. This is known as the ‘roof-tile’
effect and in simple systems allows the coupling protons to be identified. The
‘roof-tile’ effect is illustracted in Figure 37.13 can be seen in Figs. 37.8, 37.9,
and 37.11. However in molecules contain hydrogen atoms which are coupled
to several different types of proton, the ‘roof-tile’ effect is difficult to discern
and other procedures such as spin decoupling (p. 347) will be required.
5. In cases where the coupling constants between protons are not identical and
the (n + 1) rule does not apply, chemists use ‘stick diagrams’ to estimate the
nature of the coupling patterns. For example, Figure 37.14 shows the ‘stick
diagram’ for Hb of the molecular fragment ¬ CH2a ¬ CHb “ CHc ¬ X.
The coupling constants Jbc = 18 Hz and Jab = 8 Hz are applied as fol-
lows: Hb is split by Hc into a doublet (Jtrans = 18 Hz) and then each line
of the doublet is split twice by the two protons Ha (J = 8 Hz) to give a
doublet of triplets. If the CH2 is replaced by a CH3 group, a doublet of
quartets would result and a doublet of doublets if the coupling (J = 8 Hz)
was from a CH group. This is an idealised picture which can be compli-
cated by overlap of the coupled muliplets and the ‘roof-tile’ effects.
13
C9NMR spectra
The 13C nucleus has I = 12, like 1H, and the 13C ¬ NMR spectrum of a com-
pound can be observed using a different radio frequency range (in the same
magnetic field) to that for 1H. The 13C spectrum will give peaks for each
Hb
Jbc
Jab
Jab
400 350 300 250 200 150 d(ppm)

Fig. 37.14 ‘Stick diagram’ representation
1
Fig. 37.13 H9NMR spectrum of iodoethane (CH3CH2I) of coupling between protons with different
illustrating the ‘roof-tile’ effect coupling constants.

different type of carbon atom in a molecule but the properties of the 13C nucleus
give some important and useful differences in the spectrum obtained:
●● The natural abundance of 13C is only 1.1% compared with 98.9% for 12C –in
any molecule no two adjacent atoms are likely to be 13C and therefore cou-
pling between 13C nuclei will not be seen, giving a very simple spectrum.
●● In a sample of a compound, which contains many molecules, the 13C isotope
is randomly distributed and all the different carbon atoms in a sample of a
compound will be seen in the 13C ¬ NMR spectrum.
●● The sensitivity of the 13C nucleus is low and this, together with its low natu-
ral abundance, means that FT–NMR is the only practical system to produce
a spectrum by accumulation of spectra by repetitions. Larger sample size in
bigger NMR tubes also assist in solving the sensitivity/abundance problem.
●● The chemical shift range for 13C is greater (dC = 09250 ppm) than for
1
H (dH = 0915 ppm) giving greater spectral dispersion, i.e. the peaks for
carbons with very slight differences in chemical shifts are separated and do
not overlap.
13
●● C nuclei will couple with the 1H nuclei to which they are directly bonded,
e.g. CH3 will appear as a quartet, CH2 as a triplet, CH as a doublet, but C
with no hydrogen atoms attached will appear as a singlet. This introduction
of complexity in the 13C ¬ NMR spectrum is removed by broadband decou-
pling (see p. 347).
●● The peak areas of the different carbon atoms are not related to the number
of carbon atoms having the same chemical shift, as in the case for 1H9NMR
spectra.
Interpretation of 13C ¬ NMR spectra –
the spectrum is that of all the carbon Interpreting 13C9NMR spectra
atoms in the molecule. It is easy to for-
get that the peaks for carbon atoms car-
Normally you will be given two 13C9NMR spectra (Fig. 37.15). The upper
rying no hydrogen atoms are present.
spectrum, which is more complex (more lines) is called the off-resonance
decoupled spectrum and shows the 13C ¬ 1H coupling to enable you to
Definition
d c b a
Homonuclear decoupling
Application of a second radiofrequency (i)
to a NMR spectrum to remove cou-
pling between identical atomic nuclei
O
e.g. 1H9 1H decoupling.
Heteronuclear decoupling d CH3OCH2CCH3 c b a
a c db
Application of a second radiofrequency
to a NMR spectrum to remove cou-
pling between different atomic nuclei
e.g. 13C9 1H decoupling.
(ii)
f (ppm)
210.0 190.0 170.0 150.0 130.0 110.0 90.0 70.0 50.0 30.0 10.0 0.0
Fig. 37.15 13C ¬ NMR spectra of 1-methoxypropanone: (i) off-resonance

decoupled; (ii) broadband decoupled.

Ar–C sp2–C determine which carbon signals are CH3, CH2, CH and C. Then overlapping
of peaks may make the identification of different carbon atoms difficult. The
C O sp–C sp3–C
lower spectrum is a broadband decoupled spectrum in which the molecule is
irradiated with a second radio frequency range for the protons in the molecule
20 f
240 200 160 120 80 40 0
220 180 140 100 60 (heteronuclear decoupling) and effectively removes all the 13C ¬ 1H couplings
from the spectrum. The resulting simplicity of the spectrum makes identifica-
Fig. 37.16 Approximate chemical shift position of the different types of carbon in the molecule relatively easy.
tions in 13C ¬ NMR. The chemical shifts of 13C atoms (dC) vary in the same manner as those
of protons (Fig. 37.16):
1. dC moves downfield as the hybridisation of the carbon atom changes from
sp3 (0–50 ppm) to sp (75–105 ppm) to sp2 (100–140 ppm);
2. for sp3 hybridised carbon: dC moves further downfield with the change
from CH3 to CH2 to CH to C;
3. for sp2 hybridised carbon: aromatic carbons occur further downfield
(dC = 1159145 ppm) than alkene carbon atoms (dC = 1009140 ppm);
4. bonding more electronegative atoms to carbon deshields the carbon
atom and moves the peaks downfield, e.g. CH3 ¬ C (dC ∼ 6 ppm)
and CH3 ¬ O (dC ∼ 55 ppm), C “ C (dC ∼ 123 ppm) and C “ O
(dC ∼ 205 ppm).
These points are illustrated in Table 37.7. As a result of the effect of carbon
atoms along a chain, the use of 13C chemical shift correlation tables to predict
the values of dC in proposed molecular structures is more complex than for
protons. Therefore students are referred to the compilations of correlation data
and methodology provided in the texts in the ‘Sources of further study’ section
at the end of this chapter.
Techniques for the simplification of NMR spectra
As attempts to determine the structure of more complicated molecular struc-
tures have evolved, numerous methods for the simplification of complex spectra
have been developed. Three of the more useful are outlined below:
1. Homonuclear decoupling or 1H9 1H spin decoupling
In a 1H9NMR spectrum comprising multiple coupled peaks, which may be
overlapping and the ‘roof-tile’ effect not obvious, a 1H9 1H spin decoupling
13
Table 37.7 C Chemical Shift Ranges (dC) for common groups
d (ppm) d (ppm)
Alkanes 0-50 CH3 ¬ O 50-60

Alkenes 100-140 CH3 ¬ CI 10-25
Alkynes 75-105 CH3 ¬ N 15-45
Arenes 115-145 RCH2 ¬ O 35-75
Ketone C “ O 190-220 RCH2 ¬ CI 40-45
Aldehyde C “ O 185-205 RCH2 ¬ N 40-60
Acid C “ O 165-185 R2 CH ¬ O 65-90
Ester C “ O 155-180 R2 CH ¬ CI 70-80
Amide C “ O 155-180 R2 CH ¬ N 50-70
Nitrile C ‚ N 110-125 R3 C ¬ O 75-85

experiment in which a second radiofrequency is tuned to a specific chemi-

cal shift [as opposed to broadband decoupling (heteronuclear decoupling)
in 13C9NMR spectra] can be applied. The result is that any proton coupling
to the proton(s) being irradiated disappears and the coupling protons can be
identified. The selective spin decoupling of different protons in a spectrum
can yield valuable structural information and the results of the decoupling
of the protons in 4-nitrocinnamaldehyde are shown in Figure 37.17 as an
illustrative example.
e
O2N d
b
e H
C C
d
H C O
c H
a e
d
a c
10.00 9.50 9.00 8.50 8.00 7.50 7.00 6.50 6.00

(i) d(ppm)
decouple Ha
10.00 9.50 9.00 8.50 8.00 7.50 7.00 6.50 6.00

(ii) d(ppm)
decouple Hc
10.00 9.50 9.00 8.50 8.00 7.50 7.00 6.50 6.00

(iii) d(ppm)
decouple Hd
10.00 9.50 9.00 8.50 8.00 7.50 7.00 6.50 6.00

(iv)
d(ppm)
Fig. 37.17 Example of 1H9NMR spectrum simplification using homonuclear decoupling: (i) 1H9NMR spectrum of (E)-
4- nitrocinnamaldehyde; (ii) decoupling on Ha simplifies Hb showing that they are coupled; (iii) decoupling on Hc simpli-
fies Hb showing that they are coupled; (iv) decoupling on Hd simplifies He showing that they are coupled.

2. Distortionless Enhancement by Polarisation Transfer (DEPT)

When you are trying to determine the structure of a complex molecule
using 13C9NMR, one problem is to identify which signals are CH3, CH2,
CH or quaternary C where the chemical shifts and/or intensities are insuf-
ficient to make unambiguous assignments.
A useful procedure is to carry out a DEPT experiment on the broad-
band decoupled 13C spectrum. The software for the DEPT experiment is
built into most spectrometers and need not be detailed here (see Sources
for further study) but the result is commonly that CH3 and CH peaks are
displayed positively, CH2 peaks are shown negatively and quaternary C
peaks are not displayed in the DEPT spectrum.
Comparison with the original broadband decoupled spectrum allows
some definite assignments to be made. Figure 37.18 shows the effect
of a DEPT experiment on a simple molecule, 2,2,4-trimethylpentane
[(CH3)3CCH2CH(CH3)2], as an illustration of the technique.
3. COrrelation SpectroscopY (COSY)
As outlined in 1 and 2 above one of the major challenges in the structural
identification of complex molecules is to identify which nuclei are cou-
pling with each other. COSY can be used to identify proton- proton cou-
pling (1H ¬ 1H COSY) and proton – carbon coupling (1H ¬ 13C COSY).
COSY is one example of a 2-D NMR (two dimensional NMR) tech-
nique in which the radio frequency pulses and acquisition times are varied
in the spectrometer (for theory, see Sources for further study) and Fourier
Transformed into two axes at 90°. Usual NMR spectra (called a 1-D, one
(CH3)3CCH2CH(CH3)2
(ii)
CH3
CH3
CH2
CH
C (i)
60.0 50.0 40.0 30.0 20.0 10.0 0

d(ppm)
Fig. 37.18 13C9NMR spectrum of 2,2,4-trimethylpentane: (i) broadband decoupled spectrum; (ii) DEPT spectrum with
CH3 c , CH2 T, CH c and C deleted.

e gf d c b a
d ppm
O P
a b c d f
CH3CH2CH2COCH2 H
0
C C
a
H H a1 a
e g
1.0
b1 b
b
2.0
c
c
3.0
4.0
d2 d1 d
d
f2 f1 f
5.0
fg
g1 g
e
e
6.0
6.0 5.0 4.0 3.0 2.0 1.0 0 d ppm
Fig. 37.19 1H9 1H COSY spectrum of 1-propenyl butanoate.
dimensional NMR) are a plot of frequency (d) against intensity and are
displayed as a two dimensional print out.
A 2-D NMR spectrum should appear as three dimensional topograph-
ical map (or stack plot) but some peaks may be hidden behind each other
in the three dimensional plot. Instead the spectrum is viewed from above
and printed out as a contour map as shown in Figure 37.19, which is the
1
H ¬ 1H COSY spectrum of a simple molecule 1-propenyl butanoate (allyl
butyrate) CH3CH2CH2COOCH2CH “ CH2 and illustrates the key features:
●● The x- and y- axes are calibrated in frequency (chemical shift dH).
●● The spectrum of 1-propenyl butanoate is displayed along the diagonal
axis PQ.

●● The normal 1H9NMR spectrum of I-bromobutane is displayed along

both x- and y-axes so that the contours on the PQ axis can be identified.
●● The other contours not on the PQ axis are called cross peaks or off-
diagonal contours and represent the spin-coupling interactions between
coupled multiplets. Note that within complex coupling systems such as
1-propenyl butanoate, the intensity of the cross peaks can be correlated
with the magnitude of the coupling constants where varying J values are
present in the same molecule.
An illustrative procedure for the assignment of peaks and coupling protons in
a 1H9 1H COSY spectrum is shown in Box 37.2.
When you are trying to identify the structure of a complex molecule uti-
lising NMR spectroscopy, it is useful to identify which hydrogen atoms are
bonded to specific carbon atoms. In particular when there are several of the
same types of group (e.g. CH2 groups) or overlapping coupling proton mul-
tiplets. In a 1H9 13C COSY 2D experiment, the 1H9NMR spectrum is cross
correlated with the 13C9NMR spectrum enabling the protons to be assigned to
specific carbon atoms as a result of the couplings between the hydrogens and
the carbon atoms to which they are attached.
As an illustrative ex ample the 1H9 13C COSY spectrum of 1-propenyl butanoate

is shown in Figure 37.20 and you should note the following features:
●● The 13C9NMR spectrum is on the x-axis and the 1H9NMR spectrum is
on the y-axis.
Box 37.2 How to interpret a 1H9 1H@CoSy spectrum

Using the spectrum of 1-propenyl butanoate (Fig. 37.19) the CH2 group are coupling weakly with proton g of
as an example of the methodology of interpretation. the terminal CH2 group
1. Since the overall display is symmetrical about the PQ 8. Project vertically down from d2 until it intersects with
axis you need only to consider the cross peaks on the PQ axis at contour e. This shows that protons d of
one side of the PQ axis. The left hand side has been the CH2 group are coupling with proton e of the CH
selected for this exercise. group.
2. Identify the contours of the 2-D spectrum on the PQ 9. At contour f there are two cross peaks f1 (very close
diagonal axis, a, b, c, d, e, f and g. to the PQ axis) and f2 horizontally to the left.
3. At contour a, identify the cross peak a1 by scanning 10. Project vertically down from f1 until it intersects with
horizontally to the left.. the PQ axis at contour g. This shows that proton f of
the terminal CH2 group is coupling with proton g of
4. Project vertically down from a1 until it intersects with
the terminal CH2 group.
the PQ axis at contour b. This shows that protons a of
the CH3 group are coupling with protons b of the CH2 11. Project vertically down from f2 until it intersects
group. with the PQ axis at e. This shows that proton f of
the terminal CH is coupling with proton e of the CH
5. At contour b there is a cross peak b1. Project vertically
group.
down from b1 until it intersects the PQ axis at c. This
shows that protons b of the CH2 group are coupling 12. At contour g there is one cross peak g1 horizontally
with protons of the CH2 group c. to the left.
6. At contour d there are two cross peaks d1 and d2 13. Project vertically down from g1 until it intersects the
horizontally to the left. PQ axis at e. This shows that proton g of the terminal
CH2 group is coupling with proton of the CH group e.
7. Project vertically down from d1 until it intersects with
the PQ axis at contour g. This shows that protons d of

●● There is no overall spectrum on the diagonal axis as in the case of 1H9 1H

COSY spectra.
●● The contour peaks show which hydrogen atoms are attached to a spe-
cific carbon atom at the intersections of the proton and carbon chemical
shifts.
1-Propenyl butanoate contains four different CH2 groups and the spectrum
in Figure 37.20 confirms that the protons a, b, c, d, e, f and g are attached to
carbon atoms h, i, j, I, m and n respectively. Carbon atom k has no correlation
and is thus the carbon of the C “ O group.
k m n l j l h
d ppm
O
a b c d f
H
0
CH3CH2CH2COCH2
h i j k l C C a
m n
1.0
H H
e g
b
2.0
c
3.0
4.0
d
5.0
fg
e
6.0
7.0
8.0
180.0 170.0 160.0 150.0 140.0 130.0 120.0 110.0 100.0 90.0 80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0
dc ppm
Fig. 37.20 1H9 13C COSY spectrum of 1-propenyl butanoate.


Anderson, R. J., Bendell, D. J. and Groundwater, P. W. Shriner, R. L., Hermann, C. K. F., Morrill, T. C., Cur-
(2004) Organic Spectroscopic Analysis. Royal Society tin, D. Y. and Fuson, R. C. (2003) Systematic Identifi-
of Chemistry, Cambridge. cation of Organic Compounds, 8th edn. John Wiley &
Breitmaier, E. (2002) Structure Elucidation by NMR in Sons Ltd, Chichester.
Organic Chemistry: A Practical Guide, 3rd revised edn.
John Wiley & Sons Ltd, Chichester. Silverstein, R. M., Webster, F. X., Kiemle, D. J. and Bryce,
D. L. (2015) Spectroscopic Identification of Organic
Crews, P., Rodriguez, J. and Jaspars, M. (2010) Organic
Structure Analysis, 2nd edn. Oxford University Press, Chemicals, 8th edn. John Wiley & Sons Ltd, Chichester.
New York. Keeler, J. (2010) Understanding NMR Spectroscopy, 2nd
Field, L. D., Sternhell, S. and Kalman, J. R. (2013) Organic edn. John Wiley & Sons Ltd, Chichester.
Structures from Spectra, 5th edn. John Wiley & Sons Merlic, C. A., Fam, B. C. and Strouse, J. (2000) Webspectra.
Ltd, Chichester.
Available: http://www.chem.ucla.edu/~webspectra/
Gunther, H. (2013) NMR Spectroscopy: Basic Principles,
Concepts and Applications in Chemistry, 2nd edn. John
[A selection of problems, with solutions, in NMR
spectroscopy.]
Hore, P., Jones, J. and Wimperis, S. (2015) NMR: The
Toolkit - How Pulse Sequences Work. Oxford University Whittaker, D. (2000) Interpreting Organic Spectra. Royal
Press, New York. Society of Chemistry, Cambridge.
Jacobsen, N. E. (2010) NMR Spectroscopy Explained: Sim- Williams, D. and Fleming, I. (2008) Spectroscopic Methods
plified Theory, Applications and Examples for Organic in Organic Chemistry, 6th edn. McGraw-Hill, Maiden-
Chemistry and Structural Biology, 2nd edn. John
head, UK.
Pavia, D. L., Lampman, G. M., Kriz, G. S. and Vyvyan, J. A. Wolstenholme-Hogg, P. Chemtube UK.
(2015) Introduction to Spectroscopy, 5th edn. Brooks Available: https//www.youtube/com/user/chemtubeuk
Cole, Pacific Grove, California. Last accessed 14/02/2016.
Robien, W. (2016) 13C NMR of Organic Compounds 2014. [Videos of several spectroscopic and other chemistry
John Wiley & Sons Ltd, Chichester. related topics].
Study Exercises
37.1 Predict the 1H9NMR spectra of the following 37.2 Predict the 13C9NMR spectra of the following
molecules. Note: only the number of different molecules. Note: only the number of different
hydrogen atoms needs to be stated and coupling carbon atoms needs to be given and the 13C9 1H
patterns must be deduced. coupling patterns deduced.
(i) CH3CH2CH2Br (i) (CH3)3CBr
(ii) CH3CHBrCH3 (ii) CH3OCH2CH2OCH3
(iii) CH3CHBrCHO (iii) CH3CH(COOCH2CH3)2
(iv) CH3CH2OCOCH2CH2COOCH2CH3 (iv) (CH3)2C “ C(CH3)2
(v) CH3CH2OCOCH2CH2OCOCH2CH3 (v) C6H5CH3

37.3 Using the information provided in Table 37.3, 37.6 Using the information provided in Table 37.6,
estimate the d value for the protons of the CH2 estimate the chemical shift of Ha, Hb, Hc and Hd
group in: in the following molecule:
CH2 OCH3 OH
Hd NO2
O
37.4 Using the information provided in Table 37.4, Hc Ha

estimate the d value for the proton of the CH
group in: Hb
OEt 37.7 With reference to the molecule shown in

H3C C OEt Fig. 37.12f, calculate the d values for the protons
Ha, Hb and Hc to confirm that they correlate with
H
the spectrum.
37.5 Using the information provided in Table 37.5, 37.8 With reference to the molecule in 37.6
estimate the d values for the protons of the Ha above, work out the coupling patterns for the
and Hb in: protons Ha S Hd using the ‘stick diagram’
Ha
approach (p. 345) and Jortho = 10 Hz, Jmeta = 2 Hz
and Jpara = 0 Hz.
CO2H
Hb

38 Mass spectrometry
Mass spectrometry (MS) involves the bombardment of molecules, in the gas

Understanding mass spectrometry –
phase, with electrons. An electron is lost from the molecule to give a cation,
since this technique does not involve
the molecular ion (M + ), which then breaks down in characteristic ways to give
the production and measurement of
smaller fragments, which are cations, neutral molecules and uncharged radicals
electromagnetic spectra and is not
(Fig. 38.1).
based on quantum principles, it should
The mixture of molecular ion and fragments is accelerated to specific
not really be referred to as a spectro-
velocities using an electric field and then separated on the basis of their differ-
scopic technique.
ent masses by deflection in a magnetic or electrostatic field. Only the cations
are detected and a mass spectrum is a plot of mass-to-charge ratio (m/z) on the
x-axis against the number of ions (relative abundance, RA, %) on the y-axis.
Mass-to-charge ratios – in the over- A schematic of the components of a mass spectrometer is shown in Fig. 38.2
whelming majority of simple cases the and an example of a line-graph-type mass spectrum in Fig. 38.3.
ion detected is a monopositive cation; There are many types of mass spectrometer, from high-resolution dou-
thus z = 1 and the peaks seen on a ble-focusing instruments, which can distinguish molecular and fragment
low-resolution spectrometer equate to masses to six decimal places, to ‘bench-top’ machines with a quadrupole
the mass of the ion. mass detector which can resolve masses up to about m/z = 500, but only in
whole-number differences. Routinely you are most likely to encounter data
from ‘bench-top’ instruments and therefore only this type of spectrum will
be considered.
Determination of exact molecular
mass – high-resolution instruments Sample handling
enable the molecular formula of a com- For low-resolution spectra obtainable from a ‘bench-top’ MS, samples should
pound to be determined by summation be presented in the same form and quantity as demanded for gas chromato-
of the masses of the individual isotopes graphic analysis (p. 255). For high-resolution spectra contamination of any
of atoms, e.g. both ethane and metha- sort must be avoided and samples (typically less than 500 mg) should be sub-
nal have integral mass values of 30, but mitted in glass sample tubes with screwcaps containing an aluminium-foil
the accurate values are 58.046950 and insert. MS is so sensitive that the plasticisers from plastic tubes or plastic
58.010565, respectively. push-on caps will be detected, as will contaminating grease from ground-glass
joints and taps.
O O+ Mass spectra
CH3CCH2CH3 + e- CH3CCH2CH3 + 2e- The standard low-resolution mass spectrum (Fig. 38.3) is computer generated,
M+ which allows easy comparison with known spectra in a computer database for
O+
identification. The peak at the highest mass number is the molecular ion (M + ),
the mass of the molecule minus an electron. The peak at RA = 100%, the
CH3C + CH2CH3
O+ base peak, is the most abundant fragment in the spectrum and the computer
CH3CCH2CH3 automatically scales the spectrum to give the most abundant ion as 100%.
+O
The mass spectrum of a compound gives the following information about its
CH3 + CCH2CH3
chemical structure:
O+ ●● molecular ion mass, which includes information on the number of nitrogen
CH3C CH3+ + C O atoms and the presence of chlorine and bromine atoms (see p. 356) – which
is not easily obtained from IR and NMR spectra;
Fig. 38.1 Formation and fragmentation of a
molecular ion (M +). ●● the most stable major fragment (base peak), which can be correlated to the
structure of the molecule;
●● other important fragment ions, which may give information on the structure;
●● the detailed fragmentation pattern, which can be used to confirm a structure
by reference to a library database, cf. the ‘fingerprint’ region in IR spec-
trometry (p. 358).

Mass spectrometry
sample Molecular ions

molecules
injected The m/z value of the molecular ion is the summation of all the atomic
masses in the molecule, including the naturally occurring isotopes. For
organic molecules you will find a small peak (M + 1) above the apparent
electron beam vacuum pump
molecular ion mass (M + ) value due to the presence of 13C. The importance
of isotope peaks is the detection of chlorine and bromine in molecules
ion chamber
since these two elements have large natural abundances of isotopes, e.g.
35
high negative beam of Cl: 37Cl = 3:1 and 79Br: 81Br = 1:1. The mass spectra produced by mol-
potential charged ions
ecules containing these atoms are very distinctive with peaks at M + 2 and
S even M + 4 and M + 6 depending on how many chlorine or bromine atoms
N are present. The identification of the number and type of halogen atoms is
magnetic field
illustrated in Box 38.1.
ion Since the low-resolution mass spectrum produces integer values for m/z,
detector
the mass of M + indicates the number of nitrogen atoms in the molecule. If m/z
signal processor / readout
for M + is an odd integer, there is an odd number of N atoms in the molecule
and, if the value is an even number, then there is an even number of N atoms.
Fig. 38.2 Components of an electron impact

mass spectrometer.
Base peak
The molecular ion M + fragments into cations, radicals, radical cations and neu-
tral molecules of which only the positively charged species are detected. There
31
are several possible fragmentations for each M + but the base peak represents
90
the most energetically favoured process with the m/z value of the base peak
relative abundance (%)
70 representing the mass of the most abundant (and therefore most stable) posi-
tively charged species. The fragmentation of M + into the base peak follows the
50
simplified rules outlined in Box 38.2, and for a more detailed interpretation you
30
29
should consult the correlation tables to be found in the specialist texts referred
15 to at the end of the section.
10 12 14 17 33
10 20 30 40
m/z
Fragmentation patterns
The mass spectrum of a molecule is unique and can be stored in a computer.
Fig. 38.3 Mass spectrum for methanol; A match of the spectrum with those in the computer library is made in terms
m/z = mass-to-charge ratio. of molecular weight and the 10 most abundant peaks and a selection of possi-
bilities will be presented. At this point you need to correlate all the information
obtained from the spectroscopic techniques described in Chapters 29, 36, 37
Identification of isotope peaks – the and 38 together with the chemistry of the molecule to attempt to identify the
natural abundance of 13C is 1.1%. For structure of the molecule.
a molecule containing n carbon atoms When you attempt to interpret the mass spectrum remember that:
the probability is that 1.1 * n% of these
atoms will be 13C. Thus the mass spec- ●● Only the base peak is almost certain to be derived from the molecular ion.
trum of hexane (six carbons) gives a ●● Some lesser peaks may result from alternative fragmentation pathways, but
molecular ion (M +) at m/z = 86 and a these may be useful in assigning structural features.
peak at m/z = 87 (M + 1) which is 6.6%
the intensity of the M + peak. ●● MS is often used to confirm information from IR and NMR spectra; inter-
pretation of the mass spectrum alone is very difficult, except for the simplest
molecules.
Interfacing chromatography and mass spectrometry

The use and application of low-cost ‘bench-top’ mass spectrometers has
expanded in recent years. The proliferation of this type of instrument, i.e. a
chromatograph coupled to a mass spectrometer, is partly due to the lower capi-
tal cost of such instrumentation, but also to the value of the additional analytical

Mass spectrometry
Box 38.1 H
ow to identify the number of bromine or chlorine atoms in a molecule from the
molecular ion
1. Since Cl and Br have isotopes two mass numbers 50

apart, their presence in a molecule will produce peaks 90
at m/z values above M +, which are two mass numbers 80
apart, i.e. M + 2, M + 4, etc. 70
abundance
60
2. The expression for the number and intensities of 50
40 52
these peaks is given by the expansion of the formula: 30 15
20
(a + b)n 10 14 24 35 36
0
where a and b are the ratio of the two atom isotopes, 5 10 15 20 25 30 35 40 45 50 55 60
and n is the number of atoms. m/z
(a)
Example 1: If the molecule contains one chlorine
atom then: 49
90 84
(a + b)n = (3 + 1)1 = 3 + 1 80
70
Thus the mass spectrum of CH3Cl would show M + at
abundance
60 86
m/z = 50 (CH335Cl) and M + 2 at m/z = 52 (CH337Cl) 50
40
and the heights of these two peaks will be in the 51
30
approximate ratio 3:1 (Fig. 38.4(a)). 20
88
10 35 37 41
Example 2: If the molecule contains two chlorine 0
atoms then: 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95
m/z
(a + b)n = a2 + 2ab + b2 = (3 + 1)2 = 9 + 6 + 1 (b)
Thus the mass spectrum of CH2Cl2 would show M + at 108 110

m/z = 84 (CH35 35 37
2 Cl2), M + 2 at m/z = 86 (CH2 Cl Cl) 90
37
and M + 4 at m/z = 88 (CH2 Cl2) and the heights 80
of these peaks will be the approximate ratio 9:6:1 70
abundance
60
(Fig. 38.4(b)). 50
27
40
Example 3: If the molecule contains one bromine
30
atom then: 20
10 81 93
92
(a + b)n = (1 + 1)1 = 1 + 1 0
20 30 40 50 60 70 80 90 100 110
m/z
Thus the mass spectrum of CH3CH2Br would show M + (c)
at m/z = 108 (CH3CH279Br) and M + 2 at m/z = 110
(CH3CH281Br) and the heights of these peaks will be
173
in the approximate ratio 1:1 (Fig. 38.4(c)).
90
Example 4: If the molecule contains three bromine 80
70
atoms then:
abundance
60
50 175
171
(a + b)3 = a3 + 3a2b + 3ab2 + b3 = 40
30 93
(1 + 1)3 = 1 + 3 + 3 + 1 20 79
252 254
10 127 129 250 256
48
Thus the mass spectrum of CHBr3 would show 0
40 60 80 100 120 140 160 180 200 220 240 260
M + at m/z = 250 (CH79Br3), M + 2 at m/z = 252 m/z
(CH79Br81 79 81
2 Br), M + 4 at m/z = 254 (CH Br Br2) and (d)
M + 6 at m/z = 256 (CH81Br3) and the heights of
the peaks will be in the approximate ratio 1:3:3:1 Fig. 38.4 Mass spectra of: (a) CH3Cl; (b) CH2Cl2;
(Fig. 38.4(d)). (c) CH3CH2Br; (d) CHBr3.

Mass spectrometry
Box 38.2 Idealised fragmentation processes for the molecular ion (M+)
1. A@Cleavage: this involves breaking the ‘next but one bond’ to a hetero-atom (N, O, Hal, etc.) in the functional group
of a molecule. The following examples illustrate the general principles:
R CH2 Hal R CH2 Hal + R + CH2 Hal +

•
(M) (M+ ) (M-R)
R CH O R CH O+ R + CH O +
R1 H R1 H R1 H
•
(M) (M+ ) (M-R)
R C O R C O+ R + C O +
R1 R1 R1
•
(M) (M+ ) (M-R)
2. S@Bonds in alkanes: C ¬ C bonds break in preference to C ¬ H bonds and the most stable carbocation will be
formed as the base peak. For example, 2,2-dimethylpentane will give the stable (CH3 )3 C + cation as the base peak
instead of the less stable propyl cation CH3 CH2 CH2+.
CH3 CH3 CH3
CH3 C CH2CH2CH3 CH3 C + CH2CH2CH3 CH3 C+ + CH2CH2CH3
CH3 CH3 CH3

•
(M) (M+ ) (M-CH2CH2CH3)
3. Aromatic compounds: simple aromatics cleave to give a phenyl cation, m/z = 77, as the base peak which then
loses ethyne to give m/z = 51. Aromatics with CH2 next to the ring give the stable tropylium cation m/z = 91, and
then lose ethyne to m/z = 65.
X X
+ +
C4H3 +
-HC CH
•
(M) (M+ ) m/z = 77 m/z = 51
+
CH2–X CH2 •+X CH2
+ +
-HC CH
•
(M) (M+ ) m/z = 91 m/z = 65
4. B@Cleavage or McLafferty rearrangement: this is applied to molecules with a carbonyl group. If there is a hydrogen
atom on the carbon atom four away from the carbonyl oxygen (g carbon atom), a rearrangement of the molecular
ion occurs and a neutral alkene is lost from M+. This process occurs concurrently with the a@cleavage:
H
R1 H O R1 H O+ R1 +
O
g
+
b C C C
a R R R
•
(M) (M+ ) (M-R1CH=CH2)

Mass spectrometry
information that is possible. This section will consider the coupling of gas
chromatography to a mass spectrometer (GC–MS) and liquid chromatography
to a mass spectrometer (LC–MS). The key differences between the approaches
are also discussed.
Gas chromatography–mass spectrometry (GC–MS)

For specific details on capillary gas chromatography see Chapter 32. The cou-
pling of capillary GC to MS is achieved by a heated transfer line which allows
the vapour phase compounds that have been separated by the GC to remain in
the gas phase and be transported in the carrier gas, for example helium, directly
to the ion source of the MS.
Ionisation sources for GC–MS

sample The two most common approaches for ionisation of compounds in GC–MS are
inlet
ionisation chamber those based on either chemical ionisation or electron impact. The latter is the
most common. In electron impact (EI) mode, electrons produced from a heated
heated cathode tungsten or rhenium filament (cathode) are accelerated towards an anode, col-
filament anode
liding with the vapourised sample (X) and producing (positively) charged ions
(Fig. 38.5) which can be separated by MS. This can be expressed in the form
of the following equation:
lenses
X(g) + e - S X(g)
+
+ 2 e - [38.1]
Alternatively, in chemical ionisation (CI) mode, a reagent gas (e.g. methane)

mass analyser is ionised by electron bombardment to produce a molecular ion (CH4+ ). This
molecular ion then reacts with neutral methane to produce a reactant ion (CH5+ ).
Fig. 38.5 Electron impact ionisation. It is this reactant ion that interacts with the compound molecule to produce a
(positively) charged ion which can be separated by MS. The difference, in this
mode of ionisation, is that the resultant (positively) charged ion has the molecu-
lar weight of the compound plus one (i.e. XH1+ ). The chemical ionisation mode
can be expressed in the form of the following equations:
CH4 + e - S CH4+ + 2e - [38.2]
CH4+ + CH4 S CH5+ + CH3*[38.3]
X( g) + CH5+ S XH(+g) + CH4[38.4]
It is clear from the above that the CI mode is an indirect method of ionisation.
Liquid chromatography–mass spectrometry (LC–MS)

While developments in interface technology have improved considerably
in recent years, it is true to say that it went through many developments
including moving belt transport interfaces and thermospray. Modern instru-
ments have relied on the use of two approaches both of which allow ioni-
sation of compounds at atmospheric pressure and outside of the MS. The
two approaches are electrospray (ES) ionisation and atmospheric pressure
chemical ionisation (APCI).

Mass spectrometry
sample skimmer Electrospray (ES) ionisation

cone cone
A schematic diagram of the electrospray interface is shown in Fig. 38.6. In
capillary tube operation, solvent from the high-performance liquid chromatography system
is pumped through a stainless steel capillary tube, which is held at a high
potential (3–5 kV). The presence of this electric field causes the solvent to be
sprayed from the end of the capillary tube, hence the name. This action causes
highly charged solvent and solute ion droplets to be formed. Solvent from these
droplets evaporates, assisted by a flow of warm carrier gas (nitrogen). The gen-
erated ions (solvent and solute) are transported into the high-vacuum system
of the mass spectrometer via a nozzle-skimmer arrangement. The electrospray
and nozzle-skimmer arrangement are often positioned at right-angles to one
another. By allowing a potential gradient to exist between the electrospray and
nozzle-skimmer arrangement, the generated ions are ‘pulled’ into the mass
spectrometer, while at the same time allowing some discrimination between
atmospheric interface high the desirable solute ions and unwanted extraneous material, for example, salts
pressure vacuum
present in the buffer of the HPLC mobile phase.
Fig. 38.6 Electrospray ionisation.
Atmospheric pressure chemical ionisation (APCI)
A schematic diagram of the atmospheric pressure chemical ionisation inter-
Applicability of LC–MS – APCI is more face is shown in Fig. 38.7. Its operation is similar to that of the ES ionisation
focused on the analysis of low to mod- approach except in that in APCI the voltage is not applied to the stainless steel
erate molecular weight compounds capillary tubing but to a corona pin. Solvent from the HPLC system is pumped
with low to moderate polarity, while ES through a heated stainless steel capillary tube which is surrounded by a coaxial
is focused on large molecular weight flow of nitrogen gas. The combination of liquid solvent exiting the capillary
compounds of high polarity. tube and the flow of nitrogen gas produces an aerosol. Desolvation takes place
easily due to the heat applied to the solvent via the heated capillary tubing.
sample skimmer Located immediately in front of the capillary tube is the probe or corona pin
cone cone to which is applied a high voltage (2.5–3 kV). In the region around the corona
heated
capillary tube
discharge, a plasma is formed due to collisions and charge transfer processes.
Therefore this plasma will be made up of solute and solvent ions. In the same
manner as in ES ionisation the generated ions (solvent and solute) are trans-
ported into the high-vacuum system of the MS.
In ES ionisation and APCI, molecules can form singly charged ions by
loss or gain of a proton (hydrogen atom) – i.e. they can form ions repre-
sented as [M + 1]+ or [M - 1]- , where Mr = the relative molecular mass
of the compound. Therefore, it is possible to operate the mass spectrometer
in positive ion mode and determine peaks at m/z ratios of M + 1 (e.g. basic
compounds typified by amines), or in negative ion mode and determine peaks
corona pin
at m/z ratios of M - 1 (e.g. acidic compounds typified by carboxylic acids).
Both ES and APCI are regarded as ‘soft’ ionisation techniques, and as such
produce little or no fragmentation patterns. Care is also needed in the inter-
atmospheric interface high pretation of mass spectra, particularly in the presence of additives (buffer
pressure vacuum
solution) or contaminants. This is because adduct formation is possible. This
Fig. 38.7 Atmospheric pressure chemical manifests itself in the form of m/z ratios of M + 18 for ammonium adducts
ionisation or M + 23 for sodium adducts.
Understanding units – ‘amu’ or atomic Types of mass spectrometer

mass units are used to represent Mr on A variety of mass spectrometers are available for the mass/charge separation of
mass spectra. An alternative, often used charged particles. The most popular are the quadrupole mass spectrometer, ion
in the biological field, is ‘da’ or Daltons. trap mass spectrometer and the time-of-flight mass spectrometer. While each

Mass spectrometry
operates in a different manner, all are capable of separating charged particles

Definition on the basis of their m/z ratios.
Plasma – a hot, ionised gas.

Quadrupole mass spectrometer
Essentially a quadrupole mass spectrometer consists of four parallel rods
ion source detector
(Fig. 38.8). To these rods voltages (both DC and RF) are applied – different
voltages are applied to adjacent rods while opposite rods are electrically con-
nected, i.e. have the same voltages applied. By altering the applied voltages,
Fig. 38.8 Quadrupole mass spectrometer. ions of a particular m/z ratio can pass the mass spectrometer to the detector.
At the same time, other ions become unstable and are lost. By changing the
applied voltages particles of different m/z ratios can pass through the mass
spectrometer.
ion source Ion trap mass spectrometer

The ion trap mass spectrometer consists of three cylindrically symmetrical elec-
trodes to which voltages are applied. By altering the applied voltages, ions of
+ + increasing m/z ratios leave the ion trap and travel on to the detector (Fig. 38.9).
+
+
+
ring
+ electrode
+
+
+
Time-of-flight mass spectrometer
In a time-of-flight mass spectrometer, charged particles are separated according
to their velocity. Essentially, a charged particle accelerated by application of a
detector voltage has a resulting velocity that is characteristic of its m/z ratio. The ability
to separate different charged particles can be improved by increasing the flight
time of ions. This is achieved via a reflectron. It is not uncommon to find that
pre-separation of ions is achieved in a time-of-flight mass spectrometer via an
initial quadrupole mass spectrometer (Fig. 38.10).
Fig. 38.9 Ion trap mass spectrometer.
Detectors for MS
Deflection /
Injector steering The most common detector of positive ions in chromatography–mass spectrom-
plate
etry is the electron multiplier tube (EMT). The EMT (Fig. 38.11) consists of an
open aperture to which is applied a high voltage (3 kV). The positive ions are
attracted to the high negative potential of the EMT. For each positive ion that
strikes the internal surface of the EMT, the semiconductor coating produces
one electron. The generated electrons (negatively charged) are attracted to an
Reflecton
Detector area that is less negative than the open aperture. This is achieved by having the
Interface narrow end of the device earthed (zero potential). By this process the generated
electrons are drawn deeper into the EMT. On their way they strike the internal
surfaces of the device producing more secondary electrons. All the generated
Fig. 38.10 Time-of-flight mass
secondary electrons produced from the initial positive ion are collected and
spectrometer.
measured as an electric current.
Amp
M+ Data acquisition
M+
The main reason that mass spectrometry is linked to chromatographic separa-
e– tion is because of the ability of the mass spectrometer to perform quantitative
e –
analysis, and allow mass spectral data to be produced. So, while quantitative
analysis is possible for a multitude of other detectors (Chapter 32) it is an
Dynode
additional benefit for the identification of compounds in chemistry. A mass
spectrometer produces data that has time, signal intensity and spectral dimen-
Fig. 38.11 Schematic diagram of an electron sions (Fig. 38.12). Two possible data acquisition modes are possible in mass
multiplier tube. spectrometry: full scan mode and selected ion monitoring mode.

Mass spectrometry
Full scan mode

In full scan mode, ions produced by the ion source are separated with respect
Signal
to their m/z ratio by the mass spectrometer scanning the entire mass range,
u)
m
typically 0–400 amu, and a mass spectrum is recorded at the detector. In
(a
/z
this manner, all ions separated by the mass spectrometer are detected. As the
m
mass spectrometer is coupled up to a chromatographic separation system,
Time (mins) all compounds separated with respect to time are monitored in terms of a
Fig. 38.12 Schematic representation of a chromatogram – a plot of signal intensity versus time (p. 253) – and mass
typical output from a chromatography– spectral information. Depending on the method of ionisation used, it should
mass spectrometer system. be possible to identify unknown compounds by their fragmentation patterns or
by their molecular weight (see p. 356). The operator is assisted, most notably
in GC–MS, by the presence of computer databases that allow searching for
Selected Ion Monitoring – This is use- particular compounds and comparison of mass spectral information.
ful for quantitative analysis allowing
sensitive detection, e.g. PAHs in soil, a Selected ion monitoring (SIM) mode
drug in urine.
In contrast to the full scan mode, the SIM mode allows only specified ions to
be monitored. This leads to enhanced sensitivity, as the mass spectrometer is
not monitoring m/z ratios where no ions are present. The disadvantages of this
approach are that only known compounds are monitored in the chromatogram,
and no mass spectral information is available. This approach is particularly
useful once unknown compounds have been identified via the full scan mode,
leading to enhanced sensitivity and selectivity.

Anderson, R.J., Bendell, D.J. and Groundwater, P.W. Shriner, R.L., Hermann, C.K.F., Morrill, T.C., Curtin, D.Y.
(2004) Organic Spectroscopic Analysis. Royal Society and Fuson, R.C. (2004) Systematic Identification of
of Chemistry, Cambridge. Organic Compounds, 8th edn. John Wiley & Sons Ltd,
Crews, P., Rodriguez, J. and Jaspars, M. (2010) Organic Chichester.
Structure Analysis. 2nd edn. Oxford University Press, Silverstein, R.M., Webster, F.X., Kiemle, D.J. and
New York. Bryce, D.L (2015) Spectroscopic Identification of
Dass, C. (2007) Fundamentals of Contemporary Mass Organic Chemicals, 7th edn. John Wiley & Sons Ltd,
Spectrometry. John Wiley & Sons Ltd, Chichester. Chichester.
De Hoffman, E. and Stroobart,V. (2007) Mass Spec- Watson, J.T. and Sparkman, O.D. (2007) Introduction
trometry principles and Applications, 3rd edn. Wiley- to Mass Spectrometry: Instrumentation, Application
Interscience, London. and Strategies for Data Interpretation, 4th edn. John
Downword, K. (2004) Mass Spectrometry: A Foundation
Course. Royal Society of Chemistry, Cambridge. Whittaker, D. (2000) Interpreting Organic Spectra. Royal
Society of Chemistry, Cambridge.
Field, L.D., Sternhell, S. and Kalman, J.R. (2013) Organic
Structures from Spectra, 5th edn. John Wiley & Sons Williams, D. and Fleming, I. (2008) Spectroscopic Methods
Ltd, Chichester. in Organic Chemistry, 6th edn. McGraw-Hill, Maiden-
head, UK.
Pavia, D.L., Lampman, G.M., Kriz, G.S. and Vyvyan, J.A.
(2012) Introduction to Spectroscopy, 5th edn. Brooks
Cole, Pacific Grove.

Mass spectrometry
Study exercises
38.1 You are trying to identify a compound Mr = 126. 38.2 Test your knowledge of MS terminology. Explain
Infrared, 1H - and 13C - NMR spectroscopy have what the following acronyms stand for:
indicated that it is either CH3CH2 - COOCH2CH2CH3
(a) EI
or CH3CH2CH2COOCH2CH3, but have not provided
(b) CI
enough detail for a satisfactory conclusion. Pre-
(c) APCI
dict the mass spectra of the two compounds from
(d) ESI
the possible a@ and b@cleavages for each molecule
so that a distinction can be made.

39 X-ray diffraction
X-ray (powder) diffraction (XRD) is used for structure elucidation informa-

tion by identifying the crystalline phases within a specimen. For example,
XRD can differentiate between the different polymorphic forms of carbon
i.e. diamond and graphite, or different oxides of iron, e.g. FeO, Fe 2O3 and
Fe 3O4. Whereas an alternate technique, e.g. X-ray fluorescence spectroscopy
(Chapter 31), can only determine the quantity of iron present in the sample
and nothing about its chemical structure. So XRD is used to assess the crystal
structure of a solid sample.
Unit cell – A unit cell can be defined Key Point The molecules in a crystal are arranged in an
by its dimensions (atom-to-atom length orderly fashion; essentially the crystal could be cleaved to give
and associated angles). fragments that are essentially smaller version of the original
crystal sample. This process could be repeated until the basic
unit that describes a crystal is arrived at, the unit cell.
Sample – This is defined as the mate-
rial to be analysed and is representative
Various XRD instrument designs have developed over the years from the clas-
of the bulk.
sic Debye-Scherrer which used photographic film to detect the sample diffrac-
tion pattern (Fig. 39.1); X-rays enter the chamber through the collimator and
irradiate the rotating specimen. The X-rays are diffracted from the specimen
Specimen – This is the representative and detected by exposure to the photographic film. Various iterations in design
portion of the sample that is prepared of an XRD instrument have taken place such that a modern instrument would
and used in the XRD analysis. have a detector as well as a goniometer that allows angular movement of two
movable circles that contain the X-ray tube and the detector. The basic function
of the detector is to convert an X-ray photon into a measured signal. Typical
detectors for use in XRD are:
●● Proportional counter: essentially a metal container (i.e. cathode) with an
X-ray transparent window. Inside the container is an insulated wire (i.e.
anode) in an noble gas atmosphere (e.g. Ar) with a small quantity of quench-
ing gas (methane or carbon dioxide). An incoming X-ray photon interacts
with an Ar atom causing it to ionise; the argon ion is then attracted to the
cathode (container wall) and the ejected electron to the anode (wire). The
movement of charged particles results in the generation of an electric current,
that can be amplified by appropriate circuitry, and the signal recorded.
●● Scintillation screen: the screen is made by coating a surface with a thin
layer of zinc sulphide containing a trace quantity of nickel. When X-ray
photons strike the surface coating the screen emits a pale yellow light. An
alternative format is sodium iodide doped with thallium which emits in the
violet region of the visible spectrum.
●● Semi-conductor: Silicon and germanium semi-conductors produce an elec-
trical signal when they interact with X-ray radiation. The electrical signals are
generated when X-ray photons excite electrons from lower non-conducting
valence bands to the higher conduction bands in the material. These electrons
will create a pulse which is detected by an accumulation of charge in an
electric field. The silicon used for this purpose must be ultra-pure (intrin-
sic) to form either an n-type (electron-rich) or p-type (electron-poor) device.
To aid in the manufacture of this type of device, a small quantity of lith-
ium is added (to a p-type, boron-doped material). Hence the name of the

X-ray diffraction
Cones of X-ray diffraction
Film hole 2
X rays in
through
film hole 1
Direction of
Split in strip film Film
nondiffracted beam
toward top inside hole 2
camera
Film
X-ray hole 1 Center-to-center arc = 180°
beam
Fig. 39.1 Debye-Scherrer X-ray diffraction camera.
semi-conductor, a Si(Li) detector. The detector is constructed of very thin

layers of p and n silicon on opposite sides. The alternate Ge(Li) detector can
also be used.
Each detector will have its associated electronics and computer-based data
interpretation. A typical lay-out of a modern XRD instrument is shown in
Fig. 39.2. The procedure for XRD can be summarised as follows:
●● Specimen preparation. Ideally the particle size of the sample should be
approximately 10 mm. However, this is often difficult to achieve without
extensive grinding using an agate mortar and pestle. Post-grinding, the mate-
rial is dried and then packed into a specimen holder. The function of the
specimen holder is to allow maximum exposure of the specimen to the X-ray
beam with minimal exposure of the specimen mount.
●● Mounting specimen. Box 39.1 describes the ‘press and pull’ method for
mounting of the specimen. This approach can have the disadvantage of leav-
ing the upper surface of the specimen perturbed (by applying downward
pressure from the glass slide). Alternate procedures for loading of the spec-
imen are the back-loading method (Box 39.2) and the side-loading method
(Box 39.3).
●● Alternatively, crystals of small molecules can be attached with oil or glue to
a glass fibre or a loop, which is made of nylon or plastic and attached to a
solid rod. The capillary or loop is mounted on a goniometer, which allows it
to be positioned accurately within the X-ray beam and rotated.

X-ray diffraction
Substrate
Incident Thin film
X-rays
X-ray tube 2u Diffracted

X-rays
Detector
Diffracting
crystallite
d Atomic planes
Incident
X-rays u 2u
Diffracted
X-rays
Fig. 39.2 Schematic diagram of an X-ray diffraction instrument.
Box 39.1 Press and Pull method for mounting of specimen
1. Select a top-loading specimen plate (Fig. 39.3(a)), 3. Press the glass slide down until it makes contact
which is made from stainless steel, quartz glass or with the specimen plate holder; then pull the glass
plastic, with a depression in its centre. slide off to one side whilst still pressing downwards
(Fig. 39.3(c)).
2. Press the powder into the depression; flatten and
smooth the powder into the depression using a clean 4. The ground specimen is then carefully inserted into
glass slide (Fig. 39.3(b)). the XRD.
Box 39.2 Back-loading method for mounting of specimen
1. The back-loading method uses a conventional 4. Flip the specimen holder over and then remove the
top-loading specimen plate (Fig. 39.3(a)), whose back glass slide.
has been drilled out to form a ring-like plate.
5. The ground specimen is then carefully inserted into
2. Attach a glass (plain or frosted) slide to the top-side the XRD.
of the plate.
3. From the open (under-) side of the plate fill the recess
with the ground specimen; this side is then capped or
plugged.
(a) (b) (c)
Fig. 39.3 Press and pull method (a) flat plate top-loading specimen holder,
(b) specimen powder pressed into depression and (c) removal of glass slide.

X-ray diffraction
Box 39.3 Side-loading method for mounting of specimen
1. The side-loading method uses a conventional the specimen holder will aid packing of the ground
top-loading specimen plate (Fig. 39.4(a)), into which specimen.
a groove has been cut from the centre to the edge of
4. Place the specimen holder flat (with the retaining
the plate.
glass slide uppermost); then carefully remove the
2. Place a glass slide over the groove and turn such that glass slide.
the plate is on its side (Fig. 39.4(b)).
5. The ground specimen is then carefully inserted into
3. From the open (upper-) side of the plate fill the the XRD.
recess with the ground specimen; gentle tapping of
(a) (b) ●● Irradiate sample with X-rays. The instrument will need to be optimised
for a range of operating parameters including: wavelength, power and mon-
ochromatisation of the radiation, scanning range, mode of data collection
and data collection time. X-rays are generated (X-ray Tube, Fig. 39.5) by
bombarding a metal target (e.g. Cu or Mo) with a beam of electrons emitted
by a hot tungsten filament (Fig. 39.6). The incident beam will then ionise
Fig. 39.4 Side-loading method (a) side- electrons from the K-shell (1 s) of the target atom and X-rays are emitted as
loading holder, and (b) glass slide over the resultant vacancies are filled by electrons from higher energy levels e.g.
lower end of depression prior to addition from the L-level (which gives rise to Ka radiation) or M-level (which gives
of specimen. rise to Kb radiation). The generated X-rays then interact with the mounted
specimen.
cathode target ●● Detect X-ray diffraction pattern. A typical output for a NaCl powder
sample is shown in Fig. 39.7; the specimen was irradiated with X-rays
generated by Cu Ka (at 1.5418 A°). The peaks are due to X-ray diffraction
water
high e– cooling
from the different planes in the crystal (cubic) structure for NaCl i.e. the
voltage
Miller Indices.
●● Interpretation of X-ray diffraction pattern. In order to understand the
principles behind X-ray diffraction pattern interpretation it is necessary to
x-rays beryllium
window
consider the following terms and theory. When X-rays interact with the spec-
imen they produce a diffraction pattern (constructive interference) which can
be interpreted using Bragg’s Law (Fig. 39.8). For a given set of lattice planes
Fig. 39.5 X-ray tube. with an inter-plane distance of d, the condition for a diffraction (peak) to
occur can be simply written as
2d . sinu = n l
where d is the distance of separation of the planes, u is the angle of incidence,
n is an integer and l is the wavelength of the X-ray.
The diffraction pattern produced is determined by the crystal lattice. It is
possible to identify 14 unique crystal lattices; these are known as the Bra-
vais lattices (Fig. 39.9). These crystal lattice can be further refined in to
7 crystal systems (Table 39.1) with their respective atom-to-atom lengths
(a, b and c) and bond angles (a, b and g). By identifying the structure of the
unit cell allows identification of the crystal structure. Unit cells are used to
describe the smallest repeating structure in a lattice of a crystalline mate-
rial. These repeating units are what give rise to the constructive interfer-
ence defined by Bragg’s Law. Finally, the orientation of a crystal plane (or

X-ray diffraction
(a) i 200 (Cu Kc)

100
Ejected photoelectron
90 Miller indices: The peak is due to X-ray
220
80 diffraction from the {220} planes.
70
60
I 50
40
K 30 222
High speed X-ray photon 111 420
20
L 400 422 333
10 311 331
M 511
0
30 40 50 60 70 80 90
(a) ii Diffraction angle 2w (degrees)
Fig. 39.7 X-ray diffraction pattern for sodium chloride.

An electron drops
from outer shell
X-ray 1
K
L
X-ray emitted X-ray 2
M w w
(b)
i
K line d
d
c
c d
L line
Fig. 39.8 Bragg’s Law.
K
L
c
M
surface) in a lattice is specified by Miller indices. The Miller indices specify
N
not just one plane but an infinite set of equivalent planes. The application
M line
of a set of rules leads to the assignment of the Miller Indices, (hkl); a set of
Fig. 39.6 (a) Generation of X-rays numbers which quantify the intercepts and thus may be used to uniquely
(b) nomenclature of X-rays. identify the plane (or surface). Examples of the planes in a cubic crystal
system (one having a cubic unit cell with dimensions a = b = c) are shown
in Fig. 39.10. So the peaks in an X-ray diffraction pattern are directly related
to the atomic distances.
●● Report chemical structure. The International Center Diffraction Data
(ICDD) is the (international) organisation that maintains the database of
inorganic and organic spectra (http://www.icdd.com/). The database con-
tains information that includes details of d-spacing, chemical formula
and relative intensity. In addition, the RSC’s National Chemical Database
Service (http://cds.rsc.org/) (see Fig. 44.3) provides access to a range of
crystallographic databases including the Cambridge Structural Database
(CDS) System, the Inorganic Crystal Structure Database (ICSD) and
CrystalWorks. Instrument manufacturers provide fully searchable software
to aid identification.

X-ray diffraction
Body-centered Face-centered Simple Body-centered

Simple cubic
cubic (bcc) cubic (fcc) tetragonal tetragonal
Simple Body-centered Base-centered Face-centered

Rhombohedral
orthorhombic orthorhombic orthorhombic orthorhombic
Simple Base-centered
Hexagonal Triclinic
monoclinic monoclinic
Fig. 39.9 The 14 Bravais Lattices.
Table 39.1 The seven crystal systems
Lattice type Possible Bravais lattices Axis Axis angles
Cubic P, I,F a = b = c a = b = g = 90°

Tetragonal P,I a = b ≠ c a = b = g = 90°
Orthorhombic P, C, I, F a ≠ b ≠ c a = b = g = 90°
Hexagonal P a = b ≠ c a = b = 90° g = 120°
Trigonal (may be considered a P (Rhombohedral) a = b = c a = b = g 90° 6 120°
subdivision of hexagonal
Monoclinic P,C a ≠ b ≠ c a = g = 90° b 7 90°
Triclinic P a ≠ b ≠ c a ≠ b ≠ g ≠ 90°
P = primitive; C = centred; F = face@centred; I = body@centered.

X-ray diffraction
The information derived or available from powder X-ray diffraction is as

follows:
●● Lattice parameters
●● Phase identity
●● Phase purity
●● Crystallinity
●● Crystal structure
●● Percentage phase composition.
Miller Indices: (100) z
y
+
(a,0,0)
x
Miller Indices: (110) Miller Indices: (111)

z z
(0,0,a)
+
(0,a,0) (0,a,0)
+ y + y
+ +
(a,0,0) (a,0,0)
x x
Fig. 39.10 Example, Miller Indices for a cubic system.

Bennett, D. (2010) Understanding single-crystal X-ray Clegg, W. (2015) X-ray crystallography, 2nd edn. Oxford
crystallography. Wiley-VCH, London. University Press, Oxford.
Clearfield, A., Reibenspies, J.H. and Bhuvanesh, N. (2008) Ladd, M. and Palmer, R. (2013) Structure determination by
Principles and applications of powder diffraction. John X-ray crystallography. Springer, London.

X-ray diffraction
Study exercises
39.1 Sample preparation for powder XRD.

(a) What mass of powdered sample is required?
(b) Does the specimen need to be pure?
(c) Does the sample need to be packed tightly into the specimen holder?
39.2 Who was awarded the Nobel Prize (in Physics) in 1915?
39.3 The figure shows the three different X-ray diffraction patterns for Sio2.
(a) Why is there no structural information for SiO2 glass?
(b) Do quartz and cristobalite have the same structure?
Counts
4000 SiO2 Glass
2000
0
4000 Quartz
3000
2000
1000
0
4000 Cristobalite
2000
0
20 30 40 50
Position [°2Theta] (Copper (Cu))

40 Thermal analysis
Thermal methods are techniques in which changes in physical and/or chemical

properties of a substance are measured as a function of temperature. Several
methods of analysis are used:
●● Thermogravimetry (TG) is a technique in which a change in the weight of
the substance under investigation is monitored with respect to temperature
or time.
●● Differential thermal analysis (DTA) is a technique for measuring the differ-
ence in temperature between the substance under investigation and an inert
reference material with respect to temperature or time.
●● Differential scanning colorimetry (DSC) is a technique in which the energy
necessary to establish a zero temperature difference between the substance
under investigation and a reference material is monitored with respect to
temperature or time.
When carrying out a thermal analysis procedure it is important to consider and
record the following details:
●● Sample: a chemical description of the sample, plus its source and any
pre-treatment. Also, the purity, chemical composition and formula, if known.
Other important items to note are: the particle size, whether the sample has
been mixed with a ‘binder’ (and, if so, what it has been mixed with and in
what ratio) and the ‘history’ of the sample.
●● Crucible: the material and design of the sample holder is important. Obvi-
ously it is important that the crucible does not react with the sample during
Table 40.1 Common sample atmospheres
heating. In addition, the geometry of the crucible can influence the gas flow.
Thermal conductivity at ●● Rate of heating: this is very important if you intend to repeat the experiment
Gas at 1 atm 400/K (mWm −1K−1) on a subsequent occasion. Obviously the rate of heating of the sample in the
Air 33.3 crucible is not instantaneous but depends upon conduction, convection and
Carbon dioxide 25.1
radiation within the system. Thermal lag is therefore likely to be observed.
Helium 190.6 ●● Atmosphere: the nature of the atmosphere surrounding the sample is impor-
Nitrogen 32.3
tant in relation to the transfer of heat and the chemistry of the sample reac-
tion. Common sample atmospheres are shown in Table 40.1. In addition, the
flow rate of the gas is important: a static system will not remove reaction
products from the sample.
electrobalance
balance ●● Mass of sample: obviously the amount of sample will have an effect on the
control unit
heating rate. Also, sample homogeneity may be an issue with very small
gas in samples.
furnace
Thermogravimetry
programmer
sample The apparatus required for TG analysis is shown in Fig. 40.1. TG is normally
thermocouple carried out on solid samples. Example operating conditions are as follows:
Sample: calcium oxalate monohydrate
computer Crucible: platinum pan
gas out Rate: 10 K min -1
Atmosphere: nitrogen, 20 mL min -1
Fig. 40.1 Schematic diagram of a system Mass: 10.5mg
for thermogravimetry.

Thermal analysis
100 0 The result can be expressed as either a TG curve, a plot of changing weight
with respect to temperature or time, or a derivative of the curve, i.e. DTG,
residue CaO
where the first derivative of the TG curve is plotted with respect to tempera-
39
ture or time. As well as providing information on the thermal decomposition
of inorganic compounds, additional information can be deduced, e.g. sample
29.75 purity and Mr.
The oxalate hydrates of the alkaline earth metals, e.g. calcium, strontium
operating conditions and barium, are all insoluble. If a calcium salt made acidic with ethanoic acid
18.75
• Heating rate: 20°C min-1
• Sample holder: platinum
is treated with sodium oxalate solution, a white precipitate of calcium oxalate
• Atmosphere: nitrogen
12.5
START monohydrate is formed quantitatively. After washing the precipitate with eth-
• Flow rate: 10 mL min-1
anol it can be analysed. A typical TG curve, for calcium oxalate monohydrate,
CaC2O4.H2O, is shown in Fig. 40.2. Box 40.1 shows how to interpret a thermal
100 0
analysis trace for calcium oxalate monohydrate.
Fig. 40.2 A typical thermal analysis trace
for CaC2O4.H2O.
Box 40.1 How to interpret a thermal analysis trace
1. Identify the start position of the trace (see Fig. 40.2); this is usually indicated by the scale on the trace.
2. Identify any regions of decomposition: these are where there is a rapid change in the vertical axis. Three distinct
regions of decomposition can be identified in Fig. 40.2: (a) between the start and the first plateau there is a loss
of 12.5% (stage 1); (b) between the first and second plateau a loss of 18.75% occurs (stage 2); and (c) between
the second plateau and the final residue there is a loss of 29.75% (stage 3).
3. Determine the Mr of the starting material. The Mr of CaC2O4.H2O is 146.1.
4. Using the Mr determine the decomposition los associated with each region.
Stage 1: 146.1 * 12.5/100 = 18.3 (18 corresponds to the loss of water)
CaC2O4.H2O(s) S CaC2O4(s) + H2O(v)
Stage 2: 146.1 * 18.75/100 = 27.4 (28.01 corresponds to loss of CO)
CaC2O4(s) S CaCO3 (s) + CO(g)
Stage 3: 146.1 * 29.75/100 = 43.5 (44.01 corresponds to loss of CO2)
CaCO3(s) S CaO(s) + CO2(g)
5. Determination of the final product. In this case for CaC2O4.H2O the final residue is CaO (Mr = 56.08).
6. Check the Mr of the original compound
original sample (Mr) = [residue (Mr)/% residue] * 100
Residue from Fig. 40.2 is 39%. Therefore,
original sample (Mr) = [56.08/39%] * 100 = 143.8
Thus the calculated (Mr) of CaC2O4.H2O is 143.8, which is similar to the known Mr of CaC2O4.H2O of
146.1 g mol-1.
7. Assess the purity of the original material. The percentage purity of CaC2O4.H2O is calculated as follows:
143.8 * 100 * 1/146.1 = 98.43%

Thermal analysis
Applications
As well as inorganic complexes, thermal analysis is applicable to a wide range
of substances, e.g. polymers, drugs, soils and coals. It can also be applied to
mixtures of, for example, polymer blends.
Degradation of polymers
The effect of heat on polymers varies according to the type of polymer under
investigation. In an inert atmosphere, polymeric materials react in two distinct
ways: they either depolymerise or carbonise. For example, poly(methyl meth-
acrylate) may degrade back to the monomer.
Soil
The composition of soil is complex and varies with location and geology. Three
general stages of soil decomposition on heating can be identified:
1. Loss of moisture and simple organic compounds (between room temper-
ature and 150°C).
2. Ignition of soil organic matter (between 250 and 550°C).
3. Presence of minerals e.g. carbonates. The process can be complicated by
the presence of hydrated minerals, e.g. aluminium and iron oxides, and
micas (above 550°C).
Drugs
The presence of water in both ‘free’ and ‘bound’ states in pharmaceuticals
can be identified.

Gabbott, P. (ed.) (2007) Principles and Applications of Skoog, D.A., Holler, F.J. and Crouch, S.R. (2006) Prin-
Thermal Analysis. John Wiley & Sons Ltd, Chichester. ciples of Instrumental Analysis, 6th edn. Brooks/Cole,
Menczel, J.D. and Prime, R.B. (2009) Thermal Analysis Pacific Grove.
of Polymers, Fundamentals and Applications. John Wendlandt, W.W. and Collins, L.W. (1976) Thermal Anal-
Wiley & Sons Ltd, Chichester. ysis. Academic Press, Elsevier, Amsterdam.
Robinson, J.W., Skelly Frame, E.M. and Frame, G.M.
(2014) Undergraduate Instrumental Analysis, 7th edn.
Marcel Dekker, New York.
Study exercise
40.1 Use the methodology in Box 40.1 to comment on balance and gave the followingTg trace: 60–110°C,
the thermal decomposition of CuSO4.5H2O. Solid 14.4% weight loss; 120–140°C, 14.8% weight loss;
CuSO4.5H2O (1.0 mg) was heated (10 °C min -1) 240–300°C, 6.9% weight loss.
from 30°C to 400°C on a thermogravimetric

Information technology
and library resources
41. Finding and citing published information 375
42. Evaluating information 382
43. Using online resources 389
44. Internet resources 399
45. Using spreadsheets 421
46. Word processors, databases and other packages 428

41 Finding and citing published information
The ability to find scientific information is a skill required for many exercises in
Browsing in a library – this may turn up
your degree programme. You will need to research facts and published findings
interesting material, but remember the
as part of writing essays, literature reviews and project introductions, and when
books on the shelves are those not cur-
adding your lecture notes (p. 542) and revising for exams (p. 555). You must
rently out on loan. Almost by definition,
also learn how to follow scientific convention in citing source material as the
the latter may be more up-to-date and
authority for the statements you have made.
useful. To find out a library’s full holding
of books in any subject area, you need Sources of information
to search its catalogue (normally avail-
able as an online database). An on-line For essays and revision
search will also bring up electronic ver- You are unlikely to use scientific papers (primary sources, p. 388) for these
sions of textbooks (e-books) purposes – books and reviews are much more readable. If a lecturer or tutor
specifies a particular book, then it should not be difficult to find out where it
is shelved in your library, using the computerised index system. Library staff
Example The book Inorganic will generally be happy to assist with any queries. If you want to find out which
Chemistry, 4th edn, by C. Housecroft books your library holds on a specified topic, use the system’s subject index.
and A. Sharpe (2012; Prentice Hall) is You will also be able to search by author or by key words.
likely to be classified as follows: There are two main systems used by libraries to classify books: the Dewey
Decimal system and the Library of Congress system. Libraries differ in the
Dewey Decimal system:
way they employ these systems, especially by adding further numbers and
where 500 refers to natural
letters after the standard classification marks, e.g. to signify shelving position
sciences and
or edition number. Enquire at your library for a full explanation of local usage.
mathematics
The web is an ever-expanding resource for gathering both general and spe-
540 refers to chemistry
cific information (see Chapters 43 and 44). Sites fall into analogous categories
546 refers to inorganic
chemistry to those in the printed literature: there are sites with original information, sites
Library of Congress system:
that review information and bibliographic sites. One problem is that websites
may be frequently updated, so information present when you first looked may
where Q refers to science
be altered or even absent when the site is next consulted. Further, very little of
QD refers to chemistry
QD 146–197 refer to inorganic
the information on the web has been monitored or refereed. Another disadvan-
chemistry tage is that the site information may not state the origin of the material, who
wrote it or when it was written. This is considered in more detail in Box 43.5.
Examples of book classification
Dewey Library of Congress

Chemistry 540 QD1-999
Researching a new topic – reading
Analytical Chemistry 543 QD71-145
reviews (secondary sources, p. 388) can
provide you with a useful overview at Chromatography 543.544 QD117.C5; QD79.C4
the start of a new project. Crystallography 548 QD901-999
Electrochemistry 541.13 QD551-571; QD261, TP250-261
Inorganic Chemistry 546 QD146-197
Organic Chemistry 547 QD241-449
Physical Chemistry 541.1 QD450-731
Radiochemistry 541.28 QD601-655
Spectroscopy 543.42 QD95-96; QC450-467; QD272.56
Surface Chemistry 541.18 QD506-508
Synthesis (organic) 547.07 QD262

Finding and citing published information
For literature surveys and project work

Web resources – your university library
will provide you with access to a range You will probably need to consult the primary literature. If you are starting a
of web-based databases and informa- new research project or writing a report from scratch, you can build up a core
tion systems. The library Web pages will of relevant papers by using the following methods:
list these and provide links, which may
●● Asking around: supervisors or their postgraduate students will almost cer-
be worth bookmarking on your Web
tainly be able to supply you with a reference or two that will start you off.
browser. Resources especially useful to
scientists include: ●● Searching an online database, either via the Internet (see Chapter 44) or on
CD-ROM: these cover very wide areas and are a convenient way to start a
●● Web of Science, including the Sci-
reference collection, although a charge is often made for access and sending
ence Citation Index
●● IngentaConnect, including Ingenta out a listing of the papers selected (your library may or may not pass this
Medline on to you).
●● CSA Illumina ●● Consulting the bibliography of other papers in your collection – an
●● PubMed
important way of finding the key papers in your field. In effect, you are
●● ScienceDirect
taking advantage of the fact that another researcher has already done all the
●● Scopus
●● Ovid, including Cinahl and Medline hard work.
Most of these electronic resources oper- ●● Referring to ‘current awareness’ online databases: these are useful for
ate on a subscription basis and may keeping you up to date with current research; they usually provide a monthly
require an ’Athens’ username and pass- listing of article details (title, authors, source, author address) arranged by
word – for details of how to obtain these subject and cross-referenced by subject and author. Current awareness data-
consult library staff or your library’s bases cover a wider range of primary literature than could ever be available
website. in any one library. Examples include: Current Contents Connect (ISI), the
Current Advances series (Elsevier), Chemical Abstracts ACS and Analytical
Abstracts (RSC). Some online databases also offer a service whereby they
will email registered users with updates based on saved search criteria. Con-
sult library staff or your library website to see which of these databases and
services are available to you.
●● Using the Science Citation Index (SCI): this is a valuable means of explor-
ing the published literature in a given field, because it lets you see who has
cited a given paper; in effect, SCI allows you to move forward through the
literature from an existing reference. The Index is available online via ISI
Web of Science.
For specialised information

You may need to consult reference works, such as encyclopaedias, maps and
books providing specialised information. Some of this is now available on the
web, or online (consult your library’s information service web pages). Three
sources worth noting are:
●● The Handbook of Chemistry and Physics (Haynes, 2016; online via

CHEMnetBASE): the Chemical Rubber Company’s publication (affection-
ately known as the ‘Rubber Bible’) giving all manner of physical constants,
radioisotope half-lives, etc.
●● The Merck Index (O’Neil et al., 2013), which gives useful information about
organic chemicals, e.g. solubility, whether poisonous, now also available
online.
●● The Geigy Scientific Tables (8th edition), a series of six volumes (Lentner,
1981; 1984; 1986; 1990; 1992; Lentner et al., 1982) provides a wide range
of information centred on biochemistry, e.g. buffer formulae.
376 Information technology and library resources

Obtaining and organising research papers

Definitions
Obtaining a copy
Abstracts – shortened versions of It is usually more convenient to have personal copies of key research articles
papers, often those read at scientific in print or electronic format for direct consultation when working in a lab-
meetings. These may later appear as oratory or writing. The simplest way of obtaining these is to photocopy the
full papers. originals or download and/or print off copies online (e.g. as ‘.pdf’ files). For
Bibliography – a summary of the pub- academic purposes, this is normally acceptable within copyright law. If your
lished work in a defined subject area. library does not subscribe to the journal, it may be possible for them to borrow
ebook – a book published online in it from a nearby institute or obtain a copy via a national borrowing centre (an
downloadable form. ‘inter-library loan’). If the latter, you will have to fill in a form giving full
ebrary – a commercial service offering bibliographic details of the paper and where it was cited, as well as signing a
ebooks and other online resources. copyright clearance statement concerning your use of the copy. Alternatively,
eJournal – a journal published online, you might try emailing the communicating author and requesting an electronic
consisting of articles structured in the copy (‘.pdf’ file) of the article.
same way as a paper-based journal. A
valid username and password may be Organising papers
required for access.
Although the number of papers you accumulate in electronic and/or printed
Journal/periodical/serial – any publi-
formats may be small to start with, it is worth putting some thought into their
cation issued at regular intervals. In
storage and indexing before your collection becomes disorganised and unman-
biosciences, usually containing papers
ageable. Few things are more frustrating than not being able to lay your hands
(articles) describing original research
on a vital piece of information, and this can seriously disrupt your flow when
findings and reviews of literature.
writing or revising.
Monograph – a specialised book cover-
ing a single topic.
Proceedings – volume compiling written Indexing your references
versions of papers read at a scientific
meeting on a specific topic.
Whether you have obtained a printed copy, have stored downloaded files elec-
Review – an article in which recent
tronically, or have simply noted the bibliographic details of a reference, you
advances in a specific area are outlined
will need to index each resource. This is valuable for the following reasons:
and discussed. ●● You will probably need the bibliographic information for creating a reference
The primary literature – original research list for an assignment or report.
papers, published in specialist scientific
periodicals.
●● If the index also has database features, this can be useful, allowing you to
search for key words or authors.
●● If you include an ‘accession number’ and if you then file printed material
Copyright law – In Europe, copyright sequentially according to this number, it will help you to find the hard
regulations were harmonised in 1993 copy.
(Directive 93/98/EEC) to allow literary
copyright for 70 years after the death of ●● Depending on the indexing system used, you can add comments about the
an author and typographical copyright reference that may be useful at a later time, e.g. when writing an introduction
for 25 years after publication. This was or conclusion.
implemented in the UK in 1996, where,
in addition, the Copyright, Designs and
The simplest way to create an index system is to put the details on ref-
Patents Act (1988) allows the Copyright
erence cards, but database software can be more convenient and faster to sort
Licensing Agency to license institutions
once the bibliographic information has been entered. If you do not feel that
so that lecturers, students and research-
commercial software is appropriate for your needs, consider using a word
ers may take copies for teaching and
processor or spreadsheet; their rudimentary database sorting functions (see
personal research purposes – no more
Chapters 45 and 46) may be all that you require.
than a single article per journal issue,
If you are likely to store lots of references and other electronic resources
one chapter of a book, or extracts to a
digitally, then you should consider carefully how this information is kept, for
total of 5% of a book.
example, by choosing file names that indicate what the file contains and that
will facilitate sorting.

Making citations in text

Storing printed research papers – these
can easily be kept in alphabetical order It is particularly important to cite all sources of information used in your work.
within filing boxes or drawers, but if This will demonstrate to assessors that you have used appropriate source mate-
your collection is likely to grow large, it rial and it will also avoid any possibility of plagiarism (using another person’s
will need to be refiled as it outgrows the work without acknowledgement, p. 387).
storage space. You may therefore wish There are two main ways of citing articles and creating a bibliography
to add an ’accession number’ to the (also referred to as ‘references’ or ‘literature cited’).
record you keep in your database, and
file the papers in sequence according The Harvard system
to this, as they accumulate. New filing
space is only required at the end and
For each citation, the author name(s) and the date of publication are given at
you can use the accession numbers to
the relevant point in the text. The bibliography is organised alphabetically, and
form the basis of a simple cross-refer-
by date of publication for papers with the same authors. Formats normally
encing system.
adopted are, for example, ‘Smith and Jones (2005) stated that . . .’ or ‘it has been
shown that ... (Smith and Jones, 2005)’. Lists of references within parentheses
are separated by semicolons, e.g. ‘(Smith and Jones, 2005; Jones and Smith,
2009)’, normally in order of date of publication. To avoid repetition within the
same paragraph, an approach such as ‘the investigations of Smith and Jones
Using commercial bibliographic data- indicated that . . .’ could be used following an initial citation of the paper. Where
base software to organise your refer- there are more than two authors it is usual to write ‘et al.’ (or et al. if an italic
ences – for those with large numbers font is available); this stands for the Latin et alia meaning ‘and others’. If citing
of references in their collection, and more than one paper with the same authors, put, for example, ‘Smith and Jones
who may wish to produce lists of (2005; 2011)’ and if papers by a given set of authors appeared in the same year,
selected references in particular format, letter them (e.g. Smith and Jones, 2012a; 2012b).
e.g. for inclusion in a project report or
journal paper, systems like EndNote,
The numerical or Vancouver system
Reference Manager or ProCite can
reward the investment of time and Papers are cited via a superscript or bracketed reference number inserted at
money required to create a personal the appropriate point. Normal format would be, for example: ‘GC analyses4,5
reference catalogue. Appropriate bibli- have shown that . . .’ or ‘Jones [55,82] has claimed that . . .’. Repeated citations
ographic data must first be entered into use the number from the first citation. In the true numerical method (e.g. as
fields within a database (some versions in Nature), numbers are allocated by order of citation in the text, but in the
assist you to search online databases alpha-numerical method (e.g. the Annual Review series), the references are
and upload data from these). The data- first ordered alphabetically in the bibliography, then numbered, and it is this
base can then be searched and used to number which is used in the text. Note that with this latter method, adding or
create customised lists of selected ref- removing references is tedious, so the numbering should be done only when
erences in appropriate citation styles. the text has been finalised.
Key Point The main advantages of the Harvard system are

that the reader might recognise the paper being referred to and
that it is easily expanded if extra references are added. The main
Examples Incorporating references
advantages of the Vancouver system are that it aids text flow and
in text – this sample shows how you
reduces length.
might embed citations in text using
the Harvard approach:
‘. . . Smith et al. (2007) reported The ‘Katritzky’ system
the separation of PAHs using GC. A third system has been popularised in publications involving Professor Roy
However, others (Jones and Brown, Katritzky (University of Florida, USA) and uses the best features of both the
2006; Green et al., 2007) used RPHPLC. Harvard and Vancouver systems. For each reference in the text a code is writ-
In addition, Black (2008a; 2008b) ten comprising the year of publication, the journal and the page of the journal.
concluded that a mobile phase of
In the reference section the coded references are cited, together with the full
methanol:water (50:50, v/v) could sep-
arate PAHs effectively . . . ’.
reference – authors, journal, year, volume and page – in year order and a list of
journal codes in alphabetical order can be provided. For example:

in the text
Examples Incorporating references
in text – this sample shows how you . . . Robinson and Watt (34JCS1536) found . . .
might embed citations in text using
the Vancouver approach:
in the reference section
‘. . . Smith et al. reported the 34JCS1536 R. Robinson and S.J. Watt, J. Chem. Soc., 1934, 1536.
s eparation of PAHs using GC.1 The advantage of this system is that it is easy to insert missed references into
However, others used RPHPLC.2 – 3 the text, a great disadvantage of the otherwise neat Vancouver system. For
In addition, Black3,4 concluded that
details consult the reference section in Comprehensive Heterocyclic Chemistry,
a mobile phase of methanol : water
(50:50, v/v) could separate PAHs
ed. A.R. Katritzky and C.W. Rees, Pergamon, Oxford, 1984.
effectively . . . ’.
How to list your citations in a bibliography
Whichever citation method is used in the text, comprehensive details are
Examples
required for the bibliography so that the reader has enough information to
find the reference easily. Citations should be listed in alphabetical order with
Paper in journal: the priority: first author, subsequent author(s), date. Unfortunately, in terms
Smith, A. B., Jones, C.D. and
of punctuation and layout, there are almost as many ways of citing papers as
Professor, A. (2012). Innovative results
concerning our research interest.
there are journals. Your department may specify an exact format for project
Journal of New Results, 11, 234–5. work; if not, decide on a style and be consistent – if you do not pay attention
to the details of citation you may lose marks. Take special care with the fol-
Book:
lowing aspects:
Smith, A. B. (2011). Summary of my
Life’s Work. Megadosh Publishing ●● Authors and editors: give details of all authors and editors in your bibliog-
Corp., Bigcity. ISBN 0-123-45678-9. raphy, even if given as et al. in the text.
Chapter in edited book:
Jones, C. D. and Smith, A. B. (2010). ●● Abbreviations for journals: while there are standard abbreviations for the
Earth-shattering research from our titles of journals (consult library staff), it is a good idea to give the whole
laboratory. In: Research Compendium title, if possible.
1998 (ed. A. Professor), pp. 123–456.
●● Books: the edition should always be specified as contents may change
Bigbucks Press, Booktown.
between editions. Add, for example, ‘(5th edition)’ after the title of the book.
Thesis: You may be asked to give the International Standard Book Number (ISBN),
Smith, A. B. (2012). Investigations on a unique reference number for each book published.
my Favourite Topic. PhD thesis, Univer-
sity of Life, Fulchester. ●● Unsigned articles, e.g. unattributed newspaper articles and instruction man-
Website: uals: refer to the author(s) in text and bibliography as ‘Anon.’.
Jones, A.B. (2011). My Webpage on My ●● Websites: there is no widely accepted format at present. You should follow
Work in 2011. Available: http://www
departmental guidelines if these are provided, but, if these are not available,
.jonesinfo.co.uk/work2011.
Last accessed: 01/04/2012.
we suggest providing author name(s) and date in the text when using the Har-
vard system (e.g. Hacker, 2010), while in the bibliography giving the URL
Note: if your references are details in the following format: Hacker, A. (2010) University of Anytown
handwritten, you should indicate italics Homepage on Aardvarks. Available: http://www.myserver.ac.uk/homepage.
by underlining text and/or numbers. Last accessed: 01/04/2016. In this example, the web page was constructed
in 2014, but accessed in April 2016. If no author is identifiable, cite the
sponsoring body (e.g. University of Anytown, 2011), and if there is no author
or sponsoring body, write ‘Anon.’ for ‘anonymous’, e.g. Anon. (2011), and
use Anon. as the ‘author’ in the bibliography. If the web pages are undated,
either use the ‘last accessed’ date for citation and put no date after the author
name(s) in the reference list, or cite as ‘no date’ (e.g. Hacker, no date) and
leave out a date after the author name(s) in the reference list – you should be
consistent whichever option you choose.
●● Unread articles: you may be forced to refer to a paper via another without
having seen it. If possible, refer to another authority who has cited the paper,

e.g. ‘. . . Jones (2001), cited in Smith (2011), claimed that . . .’. Alternatively,
Finding dates on websites quickly –
you could denote such references in the bibliography by an asterisk and add
when visiting a particular page, you can
a short note to explain at the start of the reference list.
find occurrences of year dates begin-
ning ‘200’ by pressing the Control and ●● Personal communications: information received in a letter, seminar or
F keys together, entering 200 in the Find conversation can be referred to in the text as, for example, ‘. . . (Smith,
window that appears, then carrying out pers. comm.)’. These citations are not generally listed in the bibliography
a search using the Find next command. of papers, though in a thesis you could give a list of personal communicants
and their addresses.
●● Online material: some papers and articles are published solely online and
others are available online ahead of publication in printed form. The item
may be given a digital object identifier (DOI), allowing it to be cited and
potentially tracked before and after it is allocated to a printed issue (see
http://www.doi.org/). DOIs also allow for web page redirection by a central
Adding web references in other
systems – if you are using a differ-
agency, and CrossRef (http://www.crossref.org/) is the official DOI registra-
ent referencing system than Harvard,
tion organisation for scholarly and professional publications. DOIs can be
consult Pears and Shields (2010) or
used as ‘live’ hyperlinks in online articles, or cited in place of the volume
McMillan and Weyers (2013) for further
and page numbers for the article, with the remainder of the details cited in
advice on how to cite websites in these
the usual fashion, e.g. ‘Smith, A. and Jones, B. (2011) Our latest important
systems.
research in the form of a web-published article. Online Chemistry 8/2011
(p. 781). Published Online: 26 March 2011. DOI: 10.1083/mabi.200680019’.

Anon. CHEMnetBASE. Available: McMillan, K.M. and Weyers, J.D.B. (2013) The Study
http://www.chem netbase.com Skills Book 3rd edn. Pearson Education, Harlow.
Last accessed 01/09/2015. Neville, C. (2009) The Complete Guide to Referenc-
[Access to the Handbook of Chemistry and Physics] ing and Avoiding Plagiarism. Open University Press,
Anon. ISI Web of Science. Available: Maidenhead.
http://wos.mimas.ac.uk
O’Neil, M.J., Heckelman, P.E., Koch, C.B. and Roman,
K.J. (2013) Merck Index: An Encyclopedia of Chemi-
[Requires Athens password.]
cals, Drugs, and Biologicals, 15th edn. Merck & Co.
Grix, J. and Watkins, G. (2010) Information Skills: Find- Inc., Whitehouse Station.
ing and Using the Right Resources. Palgrave Macmillan,
Basingstoke. Pears, R. and Shields, G. (2010) Cite Them Right: The
Essential Referencing Guide, 8th edn. Palgrave
Macmillan, Basingstoke [Also available as an online
resource, see: http://www.citethemrightonline.com/].

Study exercises
41.1 Test your library skills. This exercise relies journal paper in the ‘references’ or ‘literature
on the fact that most university-level libraries cited’ section. Where used, indicate italicised
serving chemistry departments will take the text with normal underline and bold text with
scientific journal Nature. To help you to answer wavy underline. Pay attention to punctuation.
these questions, it may be beneficial to attend Compare these methods with each other, with
a library induction session if you have not done the methods recommended on pp. 382–384 of
so already. Alternatively, the library’s help or this book and with the recommendations your
enquiry desks may be able to assist you if you department or your course handbook makes.
are having problems. Are they all the same?
(a) First, find out and provide the name of the 41.3 Make website citations. Use a search engine
classification system that your university uses (p. 397) to find an informative website that cov-
for cataloguing its books and periodicals. ers each of the following:
(b) Using your library’s cataloguing system
(a) The use of SI units.
(online, preferably), find out the appropriate
(b) Description of a Diels–Alder reaction.
local classification number for the journal
(c) Melting point determination.
Nature.
(c) Where is Nature shelved in your library? Indicate how you would cite each website at the
(Your answer need refer only to most recent end of an essay (follow your department’s guide-
issues if some have been archived.) lines or use those in this chapter).
(d) What is the exact title of the landmark papers
41.4 Compare the Harvard and Vancouver methods
in the following two volumes? (i) Nature
of citation. Pair up with a partner in your class.
171, 737–738 (1953); (ii) Nature 408, 796–815
Each person should then pick one of the two
(2000).
main methods of citation and consider its pros
41.2 Explore different methods of citing refer- and cons independently. Meet together and com-
ences. Go to your library and seek out the pare your lists. Given the choice, which method
journal area for chemistry. Choose three dif- would you choose for: (a) a handwritten essay;
ferent journals in your subject area and from (b) a word-processed review; (c) an article in an
a recent edition write down how they would academic journal, and why?
print a typical citation for a multi-author
Answer to these study exercises are available at www.pearsoned.co.uk/practicalskills.

42 Evaluating information
Checking the reliability of information, assessing the relative value of differ-

Example A web search for the letters
ent ideas and thinking critically are skills essential to the scientific approach
‘PAH’ (e.g. using the search engine
Google, p. 397), will reveal that this
(p. 46). You will need to develop your abilities to evaluate information in this
acronym appears in several million way because:
websites. Not all of these deal with ●● you will be faced with many sources of information, from which you will
polycyclic aromatic hydrocarbon – need to select the most appropriate material;
the listed websites include: The
Princess Alexandra Hospital, an NHS ●● you may come across two conflicting sources of evidence and may have
hospital; The Princess Alice Hospice, to decide which is the more reliable;
in Esher, Surrey; and a healthcare
professionals site for Pulmonary ●● the accuracy and validity of a specific fact may be vital to your work;
Arterial Hypertension. These examples ●● you may doubt the quality of the information from a particular source;
are easy to identify as irrelevant
to research on ‘PAH, the group of ●● you may wish to check the original source because you are not sure
molecules’, but considering the many whether someone else is quoting it correctly.
websites that do actually cover this
topic, which might contain the exact
information you seek? How valid is Key Point Evaluating information and thinking critically
the information? Is it biased towards are regarded as higher order academic skills. The ability to
one viewpoint or hypothesis? Does it think deeply in this way is greatly valued in chemistry and will
represent current mainstream think- consequently be assessed in coursework and exam questions
ing on the topic? These are some of (see Chapter 63).
the issues that an evaluation of the
information sources might deal with.
The process of evaluating and using information can be broken down into four
stages:
1. Selecting and obtaining material. How to find sources is covered in
Chapter 41. Printed books and journals are important, but if you identify
a source of this kind there may be delays in borrowing it or obtaining a
photocopy. If the book or journal is available online, then downloading or
printing sections or papers will be more convenient and faster. The Inter-
net is often a first port of call if you wish to find something out quickly.
However, for many websites, it can be difficult to verify the authenticity
of the information given (see Box 43.5).
2. Assessing the content. You will need to understand fully what has been
written, including any technical terms and jargon used. Establish the rel-
evance of the information to your needs and assure yourself that the data
or conclusions have been presented in an unbiased way.
Definition
3. Modifying the information. In order to use the information, you may need
Plagiarism – the unacknowledged use of to alter it to suit your needs. This may require you to make comparisons,
another’s work as if it were one’s own. interpret or summarise. Some sources may require translation. Some data may
In this definition, the concept of ‘work’ require mathematical transformation before they are useful. There is a chance
includes ideas, writing, data or inven- of error in any of these processes and also a risk of plagiarism (see Box 42.1).
tions and not simply words; and the
4. Analysis. This may be your own interpretation of the information presented,
notion of ‘use’ does not only mean copy
or an examination of the way the original author has used the information.
‘word-for-word’, but also ‘in substance’
(i.e. a copy of the ideas involved). Use
of another’s work is acceptable only, if Key Point Advances in communications and information
you acknowledge the source (Box 42.1), technology mean that we can now access almost limitless
and you must use quotation marks if you knowledge. Consequently, the ability to evaluate information
are using the words of another person. has become an extremely important skill.

Evaluating information
Box 42.1 How to avoid plagiarism and copyright infringement
Plagiarism is defined on p. 386. Examples of plagiarism sources. The lecturer’s reading list or a book’s refer-
include the following ences may help you here.
●● Copying the work of a fellow student (past or present) 3. Take care when note-taking. If you decide to copy
and passing it off as your own. text word-for-word, make sure you show this clearly
in your notes with quotation marks. If you decide to
●● Using ‘essay-writing services’, such as those on offer
make your own notes based on a source, make sure
on certain websites.
these are paraphrased, rather than copy phrases from
●● Copying text or images from a source (book, journal the text. In both cases, write down full details of the
article or website, for instance) and using this within source at the appropriate point in your notes.
your own work without acknowledgement.
4. Place appropriate citations throughout your text,
●● Quoting others’ words without indicating who wrote where required. If you are unsure about when to do
or said them. this, study reviews and articles in your subject area
●● Copying ideas and concepts from a source without (see also Chapter 41).
acknowledgement, even if you paraphrase them. 5. Understand the differences between quoting, sum-
Most students would accept that some of the above marising and paraphrasing (p. 545). Apply the rele-
can only be described as cheating. However, many stu- vant conventions for text format: show clearly where
dents, especially at the start of their studies, are unaware you are quoting directly from a source by using quo-
of the academic rule that they must always acknowledge tation marks, and always cite your source(s). Try not to
the originators of information, ideas and concepts, and rely too much on quotation as this may be regarded
that not doing so is regarded as a form of academic dis- as a lack of ability to synthesise your own ideas.
honesty. If you adopt the appropriate conventions that Copyright issues are often associated with plagiarism,
avoid such accusations, you will achieve higher marks and refer to the right to publish (and hence copy) origi-
for your work as it will fulfil the marker’s expectations for nal material, such as text, images and music. Copyright
academic writing. material is indicated by a symbol © and a date (see, for
Universities have a range of mechanisms for identi example, p. iv of this book). Literary copyright is the
fying plagiarism, from employing experienced and vig- aspect most relevant to students in their academic stud-
ilant coursework markers and external examiners to ies. UK Copyright Law protects authors’ rights for life and
analysing students’ work using sophisticated software gives their estates rights for a further 70 years. Publishers
programs. Plagiarism is always punished severely when have ‘typographical copyright’ that lasts for 25 years. This
detected. Penalties may include awarding a mark of zero means that it is illegal to photocopy, scan or print out cop-
to all involved – both the copier(s) and the person whose yright material unless you have permission, or unless your
work has been copied (who is regarded as complicit in copying is limited to an extent that could be considered
the crime). Further disciplinary measures may be taken ‘fair dealing’. For educational purposes – private study or
in some instances. In severe cases, such as copying sub- research – in a scientific context, this generally means:
stantive parts of another’s work within a thesis, a student
may be dismissed from the university. ●● no more than 5% in total of a work;
If you wish to avoid being accused of plagiarism, the ●● one chapter of a book;
remedies are relatively simple:
●● one article per volume of an academic journal;
1. Make sure the work you present is always your own.
If you have been studying alongside a colleague, ●● 20% of a short book;
or have been discussing how to tackle a particular ●● one separate illustration or map.
problem with your peers, make sure you write on your
own when working on your assignments. You may only take one copy within the above limits, may
not copy for others, and may not exceed these amounts
2. Never, ever be tempted to ‘cut and paste’ from web- even if you own a copy of the original. These rules also
sites or online sources such as word processed hand- apply to web-based materials, but sometimes you will find
outs. Read these carefully, decide what the important sites where the copyright is waived. Some copying may
points are, express these in your own words and probe licensed; you should consult your library’s website or
vide literature citations to the original sources (see helpdesk to see whether it has access to licensed mate-
Chapter 41). In some cases, further investigations rial. Up-to-date copyright information is generally posted
may be required to find out details of the original close to library and departmental photocopiers.

Evaluating sources of information

One way of assessing the reliability of a piece of scientific information is to
think about how it was obtained in the first place. Essentially, facts and ideas
originate from someone’s research or scholarship, whether they are numerical
data, descriptions, concepts or interpretations. Sources are divided into two
main types:
Distinguishing between primary and
secondary sources – try the ‘IMRaD 1. Primary sources – those in which ideas and data are first communicated.
test’. Many primary sources contain The primary literature is generally published in the form of ‘papers’ (arti-
information in the order: Introduction, cles) in journals, whether printed or online. These are usually refereed
Materials and Methods, Results and by experts in the academic peer group of the author, and they will check
Discussion. If you see this format, and the accuracy and originality of the work and report their opinions back
particularly if data from an experiment, to the editors of the journal. This peer review system helps to maintain
study or observation are presented, reliability, but it is not perfect. Books and, more rarely, websites and
then you are probably reading a pri- articles in magazines and newspapers, can also be primary sources but
mary source. this depends on the nature of the information published rather than the
medium. These sources are not formally refereed, although they may be
read by editors and lawyers to check for errors and unsubstantiated or
libellous allegations.
Example If a journalist wrote an 2. Secondary sources – those which quote, adapt, interpret, translate,
article about graphene for the New
develop or otherwise use information drawn from primary sources. It is
York Times that was based on an
the act of quoting or paraphrasing that makes the source secondary, rather
article in science, the New York Times
article would be the secondary source, than the medium. Reviews are examples of secondary scientific sources,
while the science article would be the and books and magazine articles are often of this type.
primary source.
When information is modified for use in a secondary source, alterations are
likely to occur, whether intentional or unintentional. Most authors do not delib-
erately set out to change the meaning of the primary source, but they may
Taking account of the changing nature unwittingly do so, e.g. in changing text to avoid plagiarism or by oversimplifi-
of websites and wikis – by their very cation. Others may consciously or unconsciously exert bias in their reporting,
nature, these sources may change. for example, by quoting evidence that supports only one side of a debate.
This means that it is important to quote Therefore, the closer you can get to the primary source, the more reliable the
accurately from them and to give a ‘last information is likely to be. On the other hand, modification while creating a
accessed’ date when citing (see p. 383). secondary source could involve correcting errors, or synthesising ideas and
content from multiple sources.
Authorship and provenance

Clearly, much depends on who is writing the source and on what basis (e.g.
Finding out about authors and prove-
nance – these pieces of information
who paid them?). Consequently, an important way of assessing sources is to
are easy to find in most printed sources
investigate the ownership and provenance of the work (who and where it orig-
and may even be presented just below
inated from, and why).
the title, for convenience. In the case of
Can you identify who wrote the information? If it is signed or there is a
the web, it may not be so easy to find
‘by-line’ showing who wrote it, you might be able to make a judgement on
what you want. Relevant clues can be
the quality of what you are reading. This may be a simple decision, if you
obtained from ‘homepage’ links and the
know or can assume that the writer is an authority in the area; otherwise a
header, body and footer information.
little research might help (for example, by putting the name into a web search
For example, the domain may be use-
engine). Of course, just because Professor X thinks something does not make it
ful, while the use of the tilde symbol (∼ )
true. However, if you know that this opinion is backed up by years of research
in a URL usually indicates a personal,
and experience, then you might take it a little more seriously than the thoughts
rather than an institutional, website.
of a school pupil. If an author is not cited, effectively nobody is taking respon-
sibility for the content. Could there be a reason for this?

Is the author’s place of work cited? This might tell you whether the facts
Assessing substance over presentation –
or opinions given are based on an academic study. Is there a company with a
just because the information is pre-
vested interest behind the content? If the author works for a public body, there
sented well (e.g. in a glossy magazine
may be publication rules to follow and they may even have to submit their work
or a particularly well-constructed web-
to a publications committee before it is disseminated.
site), this does not necessarily tell you
much about its quality. Try to look below
Evaluating facts and ideas
the surface, using the methods men-
tioned in this chapter. However reliable the source of a piece of information seems to be, it is proba-
bly a good idea to retain a slight degree of scepticism about the facts or ideas
involved. Even information from impeccable primary sources may not be
perfect – different approaches can give different outcomes, and interpretations
can change with time and with further advances in knowledge. Table 42.1 pro-
vides a checklist for evaluating sources.
Critically examining facts and ideas is a complex task depending on the
particular issues involved, and a number of different general approaches can be
applied. You will need to decide which of the following general tips are useful
in your specific case:
Table 42.1 Checklist for assessing informa- ●● Make cross-referencing checks: triangulation – look at more than one
tion in science. How reliable is the informa- source and compare what is said in each. The cross-referenced sources
tion you have been reading. The more ‘yes’ should be as independent as possible (for example, do not compare a pri-
answers you can give below, the more mary source together with a secondary review based on it). If you find that
trustworthy you can assume it to be. all the sources give a similar picture, then you can be more confident about
the reliability of the information.
Assessing sources
●● Look at the extent and quality of citations (Chapter 41) – if references are
□□ Can you identify the author’s name? quoted, these indicate that a certain amount of research has been carried out
□□ Can you determine what relevant
beforehand, and that the ideas or results are based on genuine scholarship.
qualifications he/she holds?
□□ Can you say who employs the author? If you are doubtful about the quality of the work, these references might be
□□ Do you know who paid for the work to be worth looking at. How up to date are they? Do they cite independent work,
done? or is the author exclusively quoting their own work, or solely the work of
□□ Is this a primary or secondary source? one person?
□□ Is the content original or derived from
another source? ●● Consider the age of the source – the fact that a source is old is not neces-
sarily a barrier to truth, but ideas and facts may have altered since the date
Evaluating information of publication, and methods may have improved. Can you trace changes
through time in the sources available to you? What key events or publications
□□ Have you checked a range of sources?
□□ Is the information supported by relevant have forced any changes in the conclusions?
literature citation?
●● Try to distinguish fact from opinion – to what extent has the author sup-
□□ Is the age of the source likely to be
important regarding the accuracy of the ported a given viewpoint? Have relevant facts been quoted, via literature cita-
information? tions or the author’s own researches? Are numerical data used to substantiate
□□ Have you focused on the substance of the points used? Are these reliable and can you verify the information, for
the information presented rather than its example, by looking at the sources cited? Might the author have a reason for
packaging?
putting forward biased evidence to support a personal opinion?
□□ Is the information fact or opinion?
□□ Have you checked for any logical fallacies ●● Analyse the language used – words and their use can be very revealing.
in the arguments?
Subjective wording might indicate a personal opinion rather than an objec-
□□ Does the language used indicate anything
about the status of the information? tive conclusion. Propaganda and personal bias might be indicated by abso-
□□ Have the errors associated with any lute terms, such as ‘everyone knows . . . ’; ‘It can be guaranteed that . . . ’ or,
numbers been taken into account? a seemingly one-sided consideration of the evidence. How carefully has the
□□ Have the data been analysed using author considered the topic? A less studious approach might be indicated by
appropriate statistics?
exaggeration, ambiguity or the use of ‘journalese’ and slang. Always remem-
□□ Are any graphs constructed fairly?
ber, however, that content should be judged above presentation.

●● Look closely at any numbers – if the information you are looking at is

Analysing a graph – this process can
numerical in form, have statistical errors been taken into consideration, and,
be split into six phases:
where appropriate, quantified? If so, does this help you arrive at a conclusion
1. Considering the context and pur- about how genuine the differences are between important values?
pose of the graph.
●● Think carefully about any hypothesis-testing statistics used – are the
2. Recognising the type of presenta- methods appropriate? Are the underlying hypotheses the right ones? Have the
tion and examining the axes. results of any tests been interpreted correctly in arriving at the conclusion? To
3. Looking closely at the scale on each
deal with these matters, you will need at least a basic understanding of the ‘sta-
axis.
tistical approach’ and of commonly used techniques (see Chapters 53 and 54).
4. Examining the data presented (e.g. Interpreting data

data points, symbols, curves).
Numerical data
5. Considering errors and statistics
Information presented in public, whether as a written publication or a spo-
associated with the graph.
ken presentation, is rarely in the same form as it was when first obtained.
6. Reaching conclusions based on the Chapters 3, 47–48 deal with processes in which data are recorded and manip-
above. ulated – take particular care over percentages and proportions (p. 471), while
Chapter 53 describes the standard descriptive statistics used to ‘encapsulate’
These processes are amplified in
large data sets. Chapter 53 covers some relevant mathematical techniques. Sam-
Chapter 50.
pling (essentially, obtaining representative measurements) is at the heart of
many observational and experimental approaches in chemistry (see C hapters 5
and 6), and analysis of samples is a key component of hypothesis-testing sta-
Analysing a table – as with analysing tistics (Chapter 54). Understanding these topics and carrying out the associated
a graph, this process can be split into study exercises will help you improve your ability to interpret numerical data.
six phases:
Graphs
1. Considering the context and
Frequently, understanding and analysis in science depend on your ability to
purpose.
interpret data presented in graphical form. Sometimes, graphs may mislead
2. Examining the subheadings to see (Chapter 49). This may be unwitting, as in an unconscious effort to favour a
what information is contained in ‘pet’ hypothesis of the author. Graphs may be used to ‘sell’ a product, e.g. in
the rows and columns. advertising, or to favour a viewpoint as, perhaps, in politics. Experience in
3. Considering the units used and drawing and interpreting graphs will help you spot these flawed presentations,
checking any footnotes. and understanding how graphs can be erroneously presented (Box 49.3) will
help you avoid the same pitfalls.
4. Comparing the data values across
rows and/or down columns, look- Tables
ing for patterns, trends and unusual Tables, especially large ones, can appear as a mass of numbers and thus be more
values. daunting at first sight than graphs. In essence, however, most tables are simpler
5. Taking into account any statistics than most graphs. The construction of tables is dealt with in Chapter 50.
presented.
Critical thinking
6. Reaching conclusions based on the
above. Critical thinking involves the application of logic to a problem, or case study
issue. It requires a wide range of skills. Key processes involved include acquir-
These processes are covered in more ing and processing information; creating appropriate hypotheses and formu-
detail in Chapter 50. lating conclusions; and acting on the conclusions towards a specific objective.
Key Point Critical thinking needs reliable knowledge, but it

requires you to use this appropriately to analyse a problem. It
can be contrasted with rote learning – where you might mem-
orise facts without an explicit purpose other than building your
knowledge base.

Critical thinking is particularly important in the biosciences, because the sub-

Learning from examples – as your lec- ject deals with complex and dynamic systems. These can be difficult to under-
turers introduce you to case studies, stand for several reasons:
you will see how life scientists have
applied critical thinking to understand ●● they are often multi-faceted, involving many interactions;
the nature of biomolecules, cells and ●● it can be difficult to alter one variable in an experiment without produc-
organisms. Some of your laboratory ing confounding variables (see p. 48);
sessions may mimic the processes
involved – observation, hypothesis, ●● many variables may be unmeasured or unmeasurable;
experimental design, data gathering, ●● heterogeneity (variability) is encountered at all scales from the molecular
analysis and formulating a conclu- scale to the whole organism;
sion (see Chapter 41). These skills and
approaches can be applied in your ●● perturbation of the system can lead to unexpected (‘counter-intuitive’)
degree programme, e.g. when writing results.
about a biological issue or carrying out As a result, conclusions in biological research are seldom clear-cut. Crit-
a research project. ical thinking allows you to arrive at the most probable conclusion from the
results at hand; however, it also involves acknowledging that other conclu-
sions might be possible. It allows you to weigh up these possibilities and find
‘You can prove anything with statistics’ – a working hypothesis or explanation, but also to understand that your con-
leaving aside the issue that statistical clusions are essentially dynamic and might alter when new facts are known.
methods deal with probability, not cer- Hypothesis-testing with statistics (Chapter 54) is an important adjunct to crit-
tainty (Chapter 54), it is often possible ical thinking because it demands the formulation of simple hypotheses and
to analyse and present data in such a provides rational reasons for making conclusions.
way that they support one chosen argu- Recognising fallacies in arguments is an important aspect of critical think-
ment or hypothesis rather than another. ing. Philosophers and logicians recognise different forms of argument and
Detecting a bias of this kind can be many different fallacies in each form. Damer (2013) provides an overview of
difficult, but the critical thinking skills this wide-ranging and complex topic.
involved are essential for all scientists.
Explaining your thoughts

The context for your evaluation of the literature and the associated critical
Good writing requires good logic – thinking will normally be an essay, report or similar piece of academic writ-
understanding the logic behind what ing (Chapters 67–69). The skills involved in marshalling and explaining your
you want to write is a prerequisite for thoughts are regarded as highly important in employment and research.
creating high quality text. Creating a You may have a very specific remit, as defined in the instruction for the
plan for your writing (Chapter 69) will assignment (Chapter 70), or the topic may be open. In either case, your reading
help you to both recognise and organ- around the topic should result in an overarching position or argument you wish
ise what you want to say. to put forward – this is sometimes termed the ‘thesis’ for your writing. You may
be explaining an established viewpoint or creating an original perspective on
a topic. For both situations, the same principles apply. When setting out your
thoughts, you should:
●● make a clear statement of the issue being considered, if necessary defin-
ing terms and boundaries;
●● consider the issue from different perspectives, providing evidence for or
against different propositions;
●● cite sources of evidence and ideas that you are evaluating (Chapter 41);
●● ensure your viewpoint is logical and internally consistent;
●● structure your writing apprpriately; for example, by first considering evi-
dence for a particular view an then evidence againt it; or by considering the
development of evidence through time;

●● use an academic style of writing, avoiding personal statements (Box 69.2)

●● arrive at a conclusion, even if this is that the evidence is not sufficient to
allow firm statements to be made.
In the sciences, the norm is to use inductive reasoning – that is, to state observed
facts and assumptions at the outset, then draw a logical conclusion based on
these. The alternative, deductive reasoning, starts from a general statement,
premise or law which is held to be true and then reaches a conclusion by con-
sidering facts logically. You should look for these types of arguments in texts
and papers, and also think about possible flaws in such arguments. Chapter 64
discusses further the use of hypotheses and experiments in chemical sciences.
Text reference
Damer, T.E. (2013) Attacking Faulty Reasoning: A Prac-
tical Guide to Fallacy-Free Arguments, 7th edn. Wad-
sworth, Belmont, CA.

Currano, J. and Roth, D. (2013) Chemical Information for Van Gelder, T. Critical Thinking on the Web. Available:
Chemists. Royal Society of Chemistry, Cambridge. http://www.austhink.org/critical/
Smith, A. Evaluation of Information Sources. Available: Last accessed 01/09/15.
http://www.vuw.ac.nz/staff/alastair_smith/evaln/evaln.htm [Includes a useful directory of web resources.]
[Part of the Web-based Information Quality Virtual
Library giving details of a wide range of sources with
criteria for evaluation.]
Study exercises
42.1 Distinguish between primary and secondary liter- issues such as ‘nuclear energy’ or ‘global warm-
ature. Based on their titles and any research you ing’ would be good examples. Next, write out a
can do in your library, determine whether the fol- statement that you might use for a motion in a
lowing journals are primary or secondary sources: debate, such as ‘Is nuclear energy the solution
to our energy problems?’ or ‘Is global warming
(a) Chemical Communications
really happening?’ Then, write at least five points
(b) Analyst
in support of either side of the argument, which
(c) Organic and Biomolecular Chemistry
you should organise in tabular form. If you can
(d) Chemical Society Reviews
find more than five points, add these to your
(e) Polymer Chemistry
table, but for each point that you add to one side,
(f) Natural Product Updates
you should add one to the other side.
(g) Dalton Transactions
(h) Faraday Discussions
42.3 Analyse graphic presentations in the media. Many
(i) Analytical Abstracts
newspapers provide graphic presentations related
(j) Green Chemistry
to current issues, and graphs are frequently used
42.2 Consider a controversial issue from both sides. in television news reports. Practise critical think-
Select a current chemical topic being discussed ing skills by determining whether the graphs pre-
in the newspapers or other media. Controversial sented are a fair representation of the facts.

Information and communication technology (ICT) is vital in the modern aca-

Definitions demic world and ‘IT literacy’ is a core skill for all bioscientists. This involves
a wide range of computer-based skills, including:
Browser – a program to display web
pages and other Internet resources. ●● Accessing web pages using a ‘browser’ such as Internet Explorer, Firefox
Safari or Chrome.
FAQ – Frequently Asked Question; a file
or web page giving information on com- ●● Searching the web for useful information and resources using a search
mon queries, sometimes used as a file engine such as Google, or a meta-search engine such as Dogpile.
extension (.faq). ●● Finding what you need using online databases, such as library catalogues
FTP – File Transfer Protocol; a mecha- or complex websites, such as your university’s homepage.
nism for downloading files. ●● Downloading, storing and manipulating files.
URL – Uniform Resource Locator; the ●● Communicating via the Internet.
‘address’ for web resources.
●● Using e-learning facilities effectively.
●● Working with ‘Office’-type programs and other software (dealt with in
detail in Chapters 45 and 46).
Academic use of ICT resources – a
range of in appropriate activities will be You will probably receive an introduction to your university’s networked
identified in your university’s rules for IT systems and you will be required to follow rules and regulations that are
use of ICT systems. They may include: important for the operation of these systems. Whatever your level of experience
hacking, spamming, using another per- with PCs and the Internet, you should also follow the basic guidelines shown
son’s account, and copyright infringe- in Box 43.1. Reminding yourself of these from time to time will reduce your
ment, as well as broader aspects of chances of losing data.
behaviour covered by a code of conduct
or student charter The Internet as a global resource
The Internet is a complex network of computer networks; it is loosely organised
and no one group organises it or owns it. Instead, many private organisations,
universities and government organisations fund and operate discrete parts of it.
The web is the most popular application of the Internet. It allows easy
links to information and files which may be located on networked computers
across the world. The web enables you to access millions of ‘homepages’ or
‘websites’ – the initial point of reference with many individuals, institutions
and companies. Besides text and images, these sites may contain ‘hypertext
links’, highlighted words or phrases that take you to another Internet location
via a single mouse click.
Understanding the technology – you You can gain access to the Internet either through a network at your uni-
do not need to understand the work- versity, at most public libraries, at a commercial ‘Internet cafe’, or from home
ings of the Internet to use it – most of via a modem connected to a broadband or dial-up internet service provider
it is invisible to the user. To ensure you (e.g. Virgin Media, BT or Sky).
obtain the right facilities, you may need
to know some jargon, such as terms for
the speed of data transfer (megabits) Key Point Most material on the Internet has not been subject
and the nature of internet addresses. to peer review or vetting. Information obtained from the web or
Setting up a modem and/or local posted on newsgroups may be inaccurate, biased or spoof; do
wireless network can be complex, but not assume that everything you read is true, or even legal.
instructions are usually provided with
the hardware. White and Downs (2014) Online communication
and Gralla (2006) are useful texts if you
wish to learn more about computing
You will be allocated an email account by your university and should use
and the Internet.
this routinely for communicating with staff and fellow students, rather than
using a personal account. You may be asked to use email to submit work as

Using online resources
Box 43.1 Important guidelines for using PCs and networks
Hardware ●● Always use virus-checking programs on copied or

imported files before running them.
●● Do not drink or smoke around the computer.
●● Try not to turn the computer off more than is necessary. ●● Make backups of all important files at frequent inter-
vals (say, every 10 min), e.g. when using a word pro-
●● Never turn off the electricity supply to the machine cessor or spreadsheet.
while in use.
●● Periodically clear out redundant files.
●● Switch off the computer and monitor when not in use
(saves energy and avoids dangers of ‘hijacking’). Network rules
●● Rest your eyes at frequent intervals if working for
●● Never attempt to ‘hack’ into other people’s files.
extended periods at a computer monitor. Consult
Health and Safety Executive publications for up-to- ●● Do not give out any of your passwords to others.
date advice on working with display screens (http:// Change your password regularly. Make sure it is not
www.hse.gov.uk/msd/dse). a common word, is longer than eight characters, and
●● Never try to reformat the hard disk without the help includes numerical characters and punctuation sym-
of an expert. bols, as well as upper and lower case letters.
●● Never use network computers to access or provide
CDs and USB drives
financial or other personal information: spyware and
●● Protect CDs when not in use by keeping them in hold- Trojan programs may intercept your information.
ers or boxes.
●● Never open email attachments without knowing where
●● Label USB (Universal Serial Bus) drives with your they came from; always virus-check attachments
name and return details and consider adding these to before opening.
a file stored on the drive.
●● Remember to log out of the network when finished;
●● Try not to touch the surface of CDs, and if they need others can access your files if you forget to log out.
cleaning, do so carefully with a clean cloth, avoiding
scratching. If floppy disks are used, keep these away ●● Be polite when sending email messages.
from sources of magnetism (e.g. speakers).
●● Periodically reorganise your email folder(s). These rap-
●● Keep disks and USB drives away from moisture, idly become filled with acknowledgements and redun-
excess heat or cold. dant messages that reduce server efficiency and take
●● Do not use disks from others, unless you first check up your allocated filespace.
them for viruses. ●● Do not play games without approval – they can affect
●● Do not insert or remove a disk or USB drive when it is the operation of the system.
operating (drive light on). Close all files before remov-
●● If you are setting up your own network, e.g. in your
ing a USB drive and use the Safely Remove Hardware
flat, always install up-to-date firewall software, anti-
feature.
spyware and anti-virus programs.
●● Try not to leave a disk or USB drive in the computer
when you switch off. The Golden Rule – always make backup copies of impor-
tant files and store them well away from your working
File management copies. Ensure that the same accident cannot happen to
both copies.
●● Organise your files in an appropriate set of folders.
an attachment, or you may be asked to use a ‘digital drop-box’ within the uni-
versity’s e-learning system (Box 43.2). When using email at university, follow
conventions, including etiquette, carefully:
●● Check your email account regularly (daily). Your tutors may wish to send
urgent messages to you in this way.
●● Respond promptly to emails. Even if you are just acknowledging receipt,
it is polite to indicate that you have received and understood the message.

Box 43.2 Getting to grips with e-learning
Some key aspects of tackling e-learning are outlined below. ●● Try to gain as much as you can from formative
online assessments (p. 555). If these include feed-
1. Develop your basic IT skills, if required. e-learning
back on your answers, make sure you learn from
requires only basic IT skills, such as: use of keyboard
this and, if you do not understand it, consult your
and mouse; word processing; file management;
tutors.
browsing and searching. If you feel weak on any of
●● Learn from the critical descriptions that your lec-
these, seek out additional courses offered by the IT
turers provide of linked websites. These pointers
administration or your department.
may help you to evaluate such resources for your-
2. Visit your e-learning modules regularly. You should self in future.
try to get into a routine of doing this on a daily basis ●● Do not think that you will automatically assimilate
at a time that suits you. Staff will present up-to- information and concepts, just because you are
date information (e.g. lecture room changes) via viewing them online. The same principles apply as
the ‘announcements’ section, may post information with printed media: you must apply active learn-
about assessments, or links to the assessments them- ing methods (p. 548).
selves, and you may wish to provide feedback or look ●● Help your lecturers by providing constructive
at discussion boards and their threads. feedback when they ask for it. You may find this
easier to do online, than hurriedly filling out a
3. Participate. e-learning requires an active approach.
feedback sheet at the end of a session.
●● At the start of each new course, spend some time
getting to know what has been provided online 4. Organise files and web links. Take the time to cre-
to support your learning. As well as valuable ate a meaningful system of folder’s and files for
resources, this may include crucial information downloaded material in tandem with your own
such as learning outcomes (p. 555), dates of sub- coursework files and set up folders on your browser
mission for coursework and weighting of marks for bookmarked websites (Favorites in Internet
for different elements of the course. Explorer).
●● If you are allowed to download lecture notes (e.g. 5. Take care when submitting coursework. Make sure
in the form of PowerPoint presentations), do not you keep a backup of any file you email or submit
think that simply reading through these will be online and check the version you are sending care-
an adequate substitute for attending lectures and fully. Follow instructions carefully, for example
making further notes (see p. 542). regarding file type, or how to use your system’s ‘dig-
●● Do not be tempted to ‘lurk’ on discussion boards: ital drop-box’.
take part. Ask questions; start new threads;
answer points raised by others if you can.
●● Be polite. Email messages can seem abrupt and impersonal. Take care to
Spam, junk mail and phishing – these
read your messages through before sending and if you are at all in doubt, do
should be relatively easy to identify,
not send your message right away: re-read at a later time and consider how
and should never be responded to or
others might view what you say.
forwarded. Some may look ‘official’ and
request personal or financial details (for ●● Consider content carefully. A useful approach is only to send what you
example, they may pretend to come would be happy to hear being read out loud to classmates or family.
from your bank, and ask for account
●● Take care with language and names when communicating with tutors
details). Never send these details by
and other staff. Slang phrases and text message shorthand are unlikely to
email or your identity may be used
be understood. Be friendly without being overfamiliar.
illegally.
●● Use your university email for academic purposes – this includes dis-
cussing coursework with classmates, but not forwarding questionable jokes,
potentially offensive images, links to inappropriate websites, etc. In fact,
doing so may break regulations and result in disciplinary action.
●● Beware of spam, junk and ‘phishing’ via email.
Similar rules apply to discussion board and the learning management system.

The Usenet Newsgroup service is an electronic discussion facility, and

Using newsgroups and online
there are thousands of newsgroups representing different interests and topics.
forums – these can be useful to obtain
Any user can contribute to the discussion within a topic by posting their own
answers to a specific problem: just post
message; it is like email, but without privacy, since your message becomes
a query to the appropriate group and
available to all other subscribers.
wait for someone to reply. However,
Your university may also make use of social media, including Facebook
always bear in mind that this is the view
and Twitter. Ofter these systems will be explained during the initial orientation
of an individual person.
period, including how to ‘follow’ the university so that you can keep up to date
through news feeds.
Internet tools
The specific programs you will use for accessing the Internet will depend on
what has been installed locally, on the network you are using, and on your
Internet service provider. The best way to learn the features of the programs is
to try them out, making full use of whatever help services are available.
e-learning systems
Most university departments present their courses through a mixture of face-
to-face sessions (e.g. lectures, tutorials and practicals) and online resources
(e.g. lecture notes, websites, discussion boards, computerised tests and assess-
ments). This constitutes ‘blended learning’ on your part, with the online com-
ponent also being known as e-learning. Some courses are delivered fully online
(distance education) or by online study supplemented by block attendance on
campus (‘intensives’ or residential schools).
The e-learning element is usually delivered through an online module
within a virtual learning environment (e.g. Blackboard, Moodle). It is important
not to neglect the e-learning aspects of your course just because it may not be
as rigidly timetabled as your face-to-face sessions. This flexibility is to your
advantage, as you can work when it suits you, but it requires discipline on your
part. Box 43.2 provides tips for making the most of the e-learning components
of your courses. Mobile learning (m-learning), facilitated by wireless devices,
Definition is likely to grow in importance in the next few years.
m-learning – learning delivered via
mobile devices, including handheld Internet browsers
computers and mobile phones: m-learn- These are software programs that interact with remote server computers around
ing occurs independently of the location the world to carry out the tasks of requesting, retrieving and displaying the
of the learner. information you require. Many different browsers exist, but the most popular
are Internet Explorer, Mozilla Firefox and Google Chrome. These three brows-
ers dominate the market and have plug-ins and add-on programs available that
allow, for example, video sequences to be seen online. The standard functions
Definition of browsers include:
Bookmark – a feature of browsers that ●● accessing web documents;
allows you to save details of websites
●● following links to other documents;
you have visited. This is termed ‘add
to favorites’ in Internet Explorer. Book- ●● printing the current document;
marks save you the trouble of remem-
●● maintaining a history of visited URLs (including ‘bookmarks’ for key sites);
bering complex URL names and of
typing them into the browser’s address ●● searching for a term in a document;
window.
●● viewing images and image maps.

Browsers provide access to millions of websites. Certain sites specialise

Example – Useful web portals in providing catalogued links to other sites; these are known as portals and can
Organic chemistry – Internet resources be of enormous help when searching within a particular area of interest. Your
for recent topics, reactions and university’s library website will almost certainly provide a useful portal to cat-
chemicals at: http://www.organic- alogues and search services, often arranged by subject area, and this is often
chemistry.org/ the first port of call for electronic resources; get to know your way around this
part of the website as early as possible during your course.
When using a browser program to get to a particular page of information
on the web, all you require is the location (the web ‘address’) of that page, i.e.
Examples Common domains and the URL. Most web page URLs take the form http://, or https:// (secure), fol-
subdomains include: lowed by the various terms (domains and subdomains) that direct the system
to the appropriate site. If you do not have a specific URL in mind but wish to
.ac academic
explore appropriate sites, you will need to use a search tool with the browser.
.com commercial
.co commercial
.edu education (USA mainly) Search tools
.gov government (USA and UK) With the proliferation of information on the web, one of the main problems is
.mil military (USA only) finding the exact information you require. There are a variety of information
.net Internet-based companies services that you can use to filter the material on the network. These include:
.org organisation
●● search engines (Boxes 43.3 and 43.4);
.uk United Kingdom
●● meta-search engines;
●● subject directories;
●● subject gateways (portals).
Search engines such as Google (http://www.google.com), Bing (http://
Definition www.bing.com) and DuckDuck Go (https://duckduckgo.com) are tools
designed to search, gather, index and classify Web-based information. Search-
Search engine – a software system that ing is usually by keyword(s), although specific phrases can be defined. Many
returns hyperlinks to websites (‘hits’) search engines offer advanced searching tools such as the use of Boolean oper-
based on keywords specified by the ators to specify combinations of keywords to more precisely filter the sites.
user. Box 43.3 provides tips for refining keyword searches, while Box 43.4 provides
tips for enhanced searching with Google.
It is important to realise that each search engine will cover at most about
40% of the available sites and if you want to carry out an exhaustive search it is
necessary to use several to cover as much of the web as possible. Meta-search
engines make this easier. These operate by combining collections of search
engines. Examples include Mamma (http://www.mamma.com/) and Dogpile
(http://www.dogpile.com/).
Some useful approaches to searching include the following:
●● For a comprehensive search, use a variety of tools including search engines,
meta-search engines and portals or directories.
●● For a complex, finely specified search, employ Boolean operators and other
‘Dissecting’ a web address – you can
tools to refine your keywords as fully as possible (Box 43.3). Some search
often find out more about a particular
engines allow you to include and exclude terms or restrict by date.
site by progressively deleting sections ●● Use ‘cascading’ searching when available – this is searching within the
of the URL from the right-hand side. results of a previous search.
This will often take you to the home-
page of the organisation or company
●● Use advanced search facilities to limit your search, where possible, to the
involved.
type of medium you are looking for (e.g. graphics, video), language, sites in
a specific country (e.g. UK) or to a subject area (e.g. news only).

Box 43.3 Useful tips for using search engines
●● Keywords should be chosen with care. Try to make may prompt you with an alternative (correct) spelling.
them as specific as possible, e.g. search for ‘naph- Remember that US and UK spellings may differ (e.g.
thalene’, rather than ‘PAHs’ or ‘polycyclic aromatic sulphur/sulfur) and a search will only find hits from
hydrocarbons’. sites that use the spelling you specify.
●● Most search engines are case-insensitive. Thus ‘Nobel ●● Boolean operators (AND, OR, NOT) can be used with
Prize’ will return the same number of hits as ‘nobel some search engines to specify combinations of key-
prize’. If in doubt, use lower case throughout. words to more precisely filter the sites identified (e.g.
‘PAH AND analysis’ will find sites about methods to
●● Putting keyword phrases in double quotes (e.g. “cap-
analyse PAHs).
illary electrophoresis”) will result in a search for sites
with the phrase as a whole rather than sites with both ●● Some search engines allow ‘wildcards’ to be intro-
(all) parts of the phrase as separate words (i.e. ‘capil- duced with the symbol *. For example, this will allow
lary’ and ‘electrophoresis’ at different places within a you to specify the root of a word and include all pos-
site). This feature allows you to include common words sible endings, as with pentachloro*, which would find
normally excluded in the search, such as ‘the’. pentachlorophenol, pentachlorocyclohexene, etc. If the
search engine does not allow wildcards (e.g. Google),
●● Use multiple words/phrases plus similar words to
then you will need to be especially careful with the key-
improve your search, for example, ‘Harold Kroto buck-
words used, including all possible words of relevance.
minsterfullerene discovery’. If you can, use scientific
terms, as you are likely to find more relevant sites, e.g. ●● Numbers can be surprisingly useful in search engines.
search for the name of a particular chemical such as For example, typing in 1,1,2,2-tet will find sites con-
pentachlorophenol. cerned with organic compounds that might contain
halogens, e.g. 1,1,2,2-tetrachloroethane. If you know
●● Adding words preceded with + or − will add or
the phone number for a person, institute or company
exclude sites with that word present (e.g. ‘cancer
or the ISBN of the book, this can often help you find
treatment + cisplatin’ will search for all cancer treat-
relevant pages quickly.
ments that use cisplatin). This feature can also be used
to include common words normally excluded by the ●● If you arrive at a large site and cannot find the point
search engine. at which your searched word or phrase appears, press
Control and F together and a ‘local’ search window
●● Check that your search terms have the correct spell-
will appear, allowing you to find the point(s) where it
ing, otherwise you may only find sites with the same
is mentioned.
misspelled word. In some cases, the search engine
However well defined your search is, you will still need to evaluate the informa-
Downloading files from the Internet and tion obtained. Chapter 42 covers general aspects of this topic, while Box 43.5
emails – read-only files are often avail- provides specific advice on assessing the quality of information provided on
able as ‘pdf’ files that can be viewed by websites.
Adobe reader software (available free
from http://www.adobe.com), while Directories
other files may be presented as attach- A directory is a list of web resources organised by subject. It can usually be
ments to emails or as links from web browsed and may or may not have a search facility. Directories often con-
pages that can be opened by suitable tain better quality information than the lists produced by search engines, as
software (e.g. Microsoft Word or ‘paint’ they have been evaluated, often by subject specialists or librarians. Check your
programs like Paint Shop Pro).Take great library’s subject resources for relevant directories.
care in the latter cases as the transfer of
files can result in the transfer of asso-
ciated viruses. Always check new files Using the Internet as a resource
for viruses (especially .exe files) before
running them, and make sure your virus-
A common way of finding information on the web is by browsing or ‘surfing’.
detecting software is kept up to date.
However, this can be time-consuming; try to restrict yourself to sites known
to be relevant to the topic of interest. Some of the most useful sites are those

Box 43.4 Getting the most from Google searches
Google (http://www.google.com) has become the search ●● Find similar web pages. Simply click the Similar pages
engine of choice for millions of people, owing to its sim- option at the end of a Google search result to list other
plicity and effectiveness. However, you may be able to sites (note that these sites will not necessarily include
improve your searches by understanding its default set- the term(s) searched for).
tings and how they can be changed. ●● If a web link is unavailable, try the cached (stored)
page. Clicking on Cached at the end of a particular result
●● Download the Google toolbar to your browser. This is should take you to the stored page, with the additional
available from the Google homepage and will give you useful feature that the search term(s) will be highlighted.
quick access to the Google search facility.
●● Use the calculator function. Simply enter a calcula-
●● Understand how standard operators are used. For tion and press Enter to display the result, for example
combinations of keywords Google uses the ‘minus’ ‘10 + (2 * 4)’ returns 18. The calculator function can
operator ‘ - ’ instead of NOT (exclude) and the ‘plus’ also carry out simple interconversion of units, e.g. ‘2
operator ‘ + ’ instead of AND (include). Since Google feet 6 inches in metres’ returns 0.762 (see Box 4.1 for
usually ignores small words (‘stop words’ such as in interconversion factors between SI and non-SI units).
or the), use ‘ + ’ to include them in a search. Where no
operator is specified, Google assumes that you are ●● Try out the advanced search features. In addition to the
looking for both terms (i.e. ‘ + ’ is default). If you want standard operators these include the ability to specify
to search for alternative words, you can use ‘OR’ (e.g. the number of results per page (e.g. 50) to reduce the
sulphur OR sulfur). Google does not allow brackets use of the next button, language (e.g. English), file for-
and also ignores most punctuation marks. mat (e.g. for PDF files), recently updated web pages
(e.g. past 3 months), usage (e.g. free to use/share).
●● While wildcard truncation of words using ‘*’ is not
allowed, you can use ‘*’ to replace a whole word (or ●● Find non-text material. These include images, video
number). For example, if you type the phrase ‘a bond and maps – always check that any material you use is
is approximately * nanometres’ (UK spelling) your not subject to copyright limitations (p. 381).
results will give you results for web pages where the ●● Use Google alerts to keep up to date. This function
wildcard is replaced by a number. (http://www.google.co.uk/alerts) enables you to
●● Search for exact wording. By placing text in double receive regular updated searches by email.
inverted commas (“”), you can ensure that only web- ●● Use Google Scholar to find articles and papers. Go
sites with this exact phrasing will appear at the head to http://scholar.google.com/ and type in either the
of your search results. general topic or specific details for a particular arti-
●● Search within your results to improve the outcome. cle, e.g. author names or words from the title. Results
If your first search has produced a large number of show titles/authors of articles, with links to either the
results, use the Search within results option near the full article, abstract or citation. A useful feature is the
bottom of each page to type in a further word or phrase. Cited by . . . link, taking you to those papers that have
cited the article in their bibliography and enabling you
●● Search for words within the title of a web page. Use to carry out forward citation searching to locate more
the command intitle: to find a web page, for exam- recent papers. Also try out the advanced scholar search
ple intitle: ‘tissue culture’ returns web pages with this features to limit your search to a particular author, jour-
phrase in the title (note that phrases must always be nal, date or subject area. However, you should note that
in double speech marks, not single quotes). Google Scholar provides only a basic search facility to
●● Search within a website. Use the site: command to easily accessible articles and should not be viewed as a
locate words/phrases on a specific website, for exam- replacement for your library’s electronic journal holdings
ple site: iupac.org inorganic returns only those results and searching software. For example, if you find the title
for inorganic chemistry on the IUPAC website (www of a paper via Google Scholar you may then be able to
.iupac.org). Pressing Control and F when visiting a locate the electronic version through your own library’s
web page will give you a pop-up search window. databases, or request it via inter-library loan (p. 381).
●● Locate definitions, synonyms and spellings. The opera- Another significant limitation is that older (more cited)
tor define: enables you to find the meaning of a word. sources are typically listed first. Use ‘Advanced Search’
If you are unsure as to the spelling of a word, try each to find sources between years X and Y.
possibility. Google will usually return more results for ●● Use Google Earth to explore locations. Download from
the correct spelling and will often also prompt you http://earth.google.com/. This allows you to zoom in on
with the correct spelling (Did you mean ...?). satellite images to find locations.

Box 43.5 How to evaluate information on the web
It is often said that ‘you can find anything on the web’. ●● What is the purpose of the site and who is it
The two main disadvantages of this are, firstly, that you aimed at?
may need to sift through many sources before you find
●● Is the content relevant to the question you are trying
what you are looking for and, secondly, that the sources
to answer?
you find will vary in their quality and validity. It is impor-
tant to realise that evaluating sources is a key aspect ●● Is there any evidence of a potential conflict of interest,
of using the Internet for academic purposes, and one or bias? (Is the information comprehensive and bal-
that you will need to develop during the course of your anced, or narrowly focused?)
studies (see also Chapter 10). The ease with which you ●● Did you find the information via a subject-specific
can ‘point and click’ to reach various sources should not website (e.g. a bioscience gateway such as BIOME),
make you complacent about evaluating their information or through a more general source, such as a search
content. The following questions can help you to assess engine (e.g. Google)?
the quality of a website – the more times you can answer
‘yes’, the more credible the source is likely to be, and The above questions are similar to those that you would
vice versa: use in assessing the value of a printed resource (p. 388),
and similar criteria should be applied to web-based infor-
Authority mation. You should be especially wary of sites containing
●● Is the author identified? unattributed factual information or data whose primary
source is not given.
●● Are the author’s qualifications or credentials given?
●● Is the owner, publisher or sponsoring organisation Currency
identified? ●● Is there any indication that the information is up to
●● Is an address given (postal and/or email)? date?
●● Does the site show the date on which it was created,
It is sometimes possible to obtain information on
or last updated?
authority from the site’s metadata (try the ‘View’ ‘Source’
option in Internet Explorer, or look at the URL to see if it Presentation
gives any clues as to the organisation, e.g. whether the
domain name ends in .ac, .edu, .gov or .org, rather than ●● What is your overall impression of how well the site
.co or .com). has been put together?
Content ●● Are there any grammatical or spelling mistakes?
●● Is there any evidence that the information has been ●● Are there links to other websites, to support state-
peer-reviewed (p. 608), edited or otherwise validated, ments and factual information?
or is it based on such sources? The care with which a site has been constructed can
●● Is the information factual or based on personal give you an indication of the credibility of the author/
opinion? organisation. However, while a poorly presented site
may cause you to question the credibility of the informa-
●● Is the factual data original (primary) or derived from tion, the reverse is not always necessarily true: do not be
other sources (secondary)? taken in by a slick, well presented website – authority,
●● Are the sources of specific factual information detailed content and currency are always more important than
in full (Chapter 42)? presentation.

that provide hypertext links to other locations. Some other resources you can
Understanding the impermanence of
use on the web are:
the web – the temporary nature of
much of the material on the web is a ●● Libraries, publishers and commercial organisations. Your university
disadvantage for academic purposes library is likely to subscribe to one or more databases providing access to
because it may change or even disap- scientific articles; these include Web of Science (http://webofknowledge.
pear after you have cited it. You may com), and Science Direct (http://www.sciencedirect.com). A password is
also find it difficult or impossible to find usually required, especially for off-campus use; consult your library staff
out who authored the material. A case for further details. Some scientific database sites give free access, without
in point are wikis, such as Wikipedia subscription or password; these include the PubMed website of the National
(www.wikipedia.org). This online ency- Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov) and
clopaedia has many potential authors the HighWire Press (http://highwire.com). Others allow free searching, but
and the content may change rapidly as require payment for certain articles, e.g. the Scientific World (http://www.
a result of new submissions or edits; thescientificworld.com/) and Infotrieve (http://www4.infotrieve.com). Pub-
nevertheless, it can sometimes be a lishers such as Pearson and booksellers such as Amazon.com provide online
useful resource for general information catalogues and ecommerce sites that can be useful sources of information
about a wide range of topics, though it (see http://pearsoned.com and http://www.amazon.com).
is not necessarily the best approach for
●● Online journals and ebooks. A number of traditional journals have web-
researching scientific assignments.
sites. You can keep up to date by visiting the websites of Nature (http://
www.nature.com), New Scientist (http://www.newscientist.com) and Scien-
tific American (http://www.sciam.com), or ScienceDirect journals (at http://
Using traditional sources – remember
www.sciencedirect.com/science/journals). Some scientific societies make
that using the Internet to find informa-
their journals and other publications available via their websites, e.g. the.
tion is not a substitute for visiting your
Royal Society of Chemistry at http://www.rsc.org. Journals solely published
university library. Internet resources
in electronic format are also available (e.g. Molecular Vision, http://www.
complement rather than replace more
molvis.org/molvis) but some require a subscription password for access;
traditional printed sources.
check whether your institute is a subscriber. The directory of open access
jourals (at: http://www.doaj.org/) provides a comprehensive list of all
Remembering useful websites – create non-subscription (free, open access) journals.
a ‘bookmark’ ( = add a ‘favourite’) for ●● Data and images. Archives of text material, video clips and photographs
the ones you find of value, to make can be accessed, and much of the material is readily available. The HEA
revisiting easy. This can be done from Physical Sciences Centre (http://heacademy.ac.uk/physsci/) is a good exam-
the menu of your browser program. ple. When downloading such material, you should check that you are not
Make a copy of your bookmark file breaching copyright and avoid potential plagiarism by giving a full citation
occasionally, to avoid loss of this of the source if you use such images in an assignment (see p. 386).
information.
●● Chemical institutions. Many culture collections, scientific societies and
other biological institutions around the world are now online. Use their sites
Note that URLs may change – make a to obtain specific information about collections, resources, etc. They fre-
keyword search using a search engine quently provide lists of other relevant sites or topics, e.g. the Royal Society
to find a particular site if the URL infor- of Chemistry at (http://www.rsc.org).
mation you have does not lead you to
an active page. Alternatively, use the
‘Way-back Machine’ at: http://www.
archive.org to locate an earlier version
of a URL.

●● Databases. In addition to those covering the scientific literature (e.g. Web

Understanding open access publica-
of Science, or the PubMed service of the US National Library of Medicine),
tion – this is a relatively recent move-
others focus on specific topics (e.g. academic employment, http://www.
ment, pioneered by the Public Library
jobs.ac.uk). Chapter 44 provides further guidance on using online chemis-
of Science (PLOS). Typically, authors of
try databases.
scholarly articles (or their institutions)
pay for all costs required for online
publishing, thereby providing free and
unrestricted online access to all poten-
tial readers (for an example of an open
access journal, see: http://journals.plos.
org/plosone/).
Text references
Gralla, P. (2006) How the Internet Works, 8th edn. Pearson, White, R. and Downs, T. (2014) How Computers Work,
Harlow. 10th edn. Que Publishing, Pearson, Harlow.

Currano, J. and Roth, D. (2013) Chemical Information for Hock, R. (2013) The Extreme Searcher’s Internet Hand-
Chemists. Royal Society of Chemistry, Cambridge. book A Guide for the Serious Searcher, 4th edn. Cyber
Grassian, E. Thinking Critically about World Wide Age Books, New Jersey.
Web Resources. Available:
http://www.library.ucla.edu/libraries/college/
11605_12337.cfm
Study exercises
43.1 Explore the resources of the web using a search sure that you include meta-search engines such
engine. Using a search engine, find the answers as Dogpile. Compare the outcomes to reveal the
to the following questions: strengths and weaknesses of the individual search
(a) Who is Linus C. Pauling, and what is he engines. Work with a colleague to compare differ-
famous for? What prize did he win and when? ent searches on a quantitative (i.e. number of hits)
(b) What is the alternative name for and qualitative (quality of hits) basis.
dichloro-methane? 43.3 Organise your bookmarks. Enter the ‘Organise
(c) What is the postal address of the Royal Soci- favorites’ menu for your preferred browser and
ety of Chemistry’s London headquarters? create folders with appropriate headings. Move
43.2 Compare results from a variety of search existing bookmarks to these folders and save any
engines. First, think of an appropriate chemis- new ones appropriately. Doing this will help you
try keyword or phrase (e.g. a compound name) find bookmarks more easily, rather than search-
and enter this into several search engines. Make ing through long lists.

44 Internet resources
The extensive development in the availability of chemical resources on the

internet continues. One way to find such material on the Internet is by brows-
ing (‘surfing’) the web. However, as this can be time-consuming and wasteful,
browsing should be focused on relevant and appropriate sites. Many of the
most useful web sites are those providing detailed lists and hypertext links to
other locations.
Using Internet addresses – note that Key Point Remember that the Internet should not be viewed
the locations given in the chapter may as a substitute for your own University library and other local
change as sites are updated: you can resources, but should complement, rather than replace, more
make a keyword search using a search- traditional printed texts and other electronic material.
ing system or ‘search engine’ to find a
particular website if necessary.
General information
Some of the principal resources you can utilise via the web are:
Revisiting websites – ‘bookmark’ sites
●● Libraries, publishers and companies. These organisations recognise the
of interest, so that you can return to
significance of the Internet as a means of communication. For example,
them at a later date. Make a copy of
the Pearson higher education website at https://heuk.pearson.com/ allows
your bookmark file occasionally, to
information on specific catalogues and books to be requested online. A
avoid loss of relevant information.
large number of scientific journals are also available in electronic format
via ingenta.com (http://www.ingentaconnect.com). Ingenta journals offers
a single point of access to over 10 000 academic and professional journals.
Access to browse and search the database of articles is free, as is the ability to
Definition:
display tables of contents, bibliographic information and abstracts. Full-text
RSS – Really Simple Syndication
articles require a fee for access – check whether your institution subscribes.
A broad range of scientific journals are also published by Elsevier and are
available via the site http://www.elsevierdirect.com/index.jsp. You can keep
up to date using New Scientist pages (at http://www.newscientist.com/),
Blog – a contraction of the words ‘web Scientific American (at http://www.scientificamerican.com/) or Nature (at
log’; it is essentially a type of website, http://www.nature.com/). Table 44.1 shows a selected list of journals that
maintained by an individual, who can would be of interest to a synthetic organic chemist.
update commentary, videos or graphics
●● Institutions. Many research organisations, societies and educational institutions
on to the site.
around the world are now online, with their own web pages. There is a detailed
list of scholarly chemical societies at http://www.chemdex.org/. You can use the
websites of these organisations to obtain specific information. They frequently
provide hypertext links to other relevant sites for particular groups or topics: for
example, the Royal Society of Chemistry web page (http://www.rsc.org) has
links to various sites of interest to chemists. Use the web to obtain details of the
activities of research organisations (e.g. GlaxoSmithKline, at http://www.gsk
.com/) or individual laboratories and researchers at specific universities. Some
other relevant websites are given in Table 44.2.
●● Data and pictures. Archives of text material, photographs and video clips
can be accessed and easily downloaded. However, downloading images may
take quite a while, especially for remote links or at busy times.
●● Newsgroups (Usernet). News articles on a wide range of topics are ‘posted’
at appropriate sites, where they are placed into subject groups (newsgroups).
Any user can contribute to the discussion by posting his/her own message,

Internet resources
Table 44.1 Selected online chemistry journals Table 44.1 Continued
Journal Publisher Journal Publisher
Accounts of Chemical American Chemical Society Heterocycles Japan Institute of

Research Heterocyclic Chemistry
Acta Crystallographica (A-F) International Union of Inorganic Chemistry American Chemical Society
Crystallography
Journal of the American American Chemical Society
Angewandte Chemie Interna- Wiley-VCH Chemical Society
tional Edition
Journal of Combinatorial American Chemical Society
Bioorganic and Medicinal Elsevier Chemistry
Chemistry
Journal of Medicinal American Chemical Society
Bioorganic and Medicinal Elsevier Chemistry
Chemistry Letters
Journal of Molecular Cataly- Elsevier
Bulletin of the Chemical Soci- Chemical Society of Japan sis A: Chemical
ety of Japan
Journal of Molecular Cataly- Elsevier
Carbohydrate Research Elsevier sis B: Enzymatic
Carbon Elsevier Journal of Natural Products American Chemical Society
ChemBioChem Wiley-VCH Journal of Organic Chemistry American Chemical Society
Chemical Communications Royal Society of Chemistry Journal of Organometallic Elsevier
Chemistry
Chemical & Engineering News American Chemical Society
Natural Product Reports Royal Society of Chemistry
Chemical Reviews American Chemical Society
Nature Nature Publishing Group
Chemical Society Reviews Royal Society of Chemistry
New Journal of Chemistry Royal Society of Chemistry
Chemistry: A European Journal Wiley-VCH
Organic Letters American Chemical Society
Chemistry & Biology Elsevier
Organic Syntheses Wiley
Chemistry Letters Chemical Society of Japan
Organometallics American Chemical Society
Chemistry of Materials American Chemical Society
Perkin Transactions 1 Royal Society of Chemistry
European Journal of Inor- Wiley-VCH
ganic Chemistry Phytochemistry Elsevier
European Journal of Elsevier Pure & Applied Chemistry IUPAC
Medicinal Chemistry
Russian Chemical Reviews Royal Society of Chemistry
European Journal of Organic Wiley-VCH
Science AAAS
Chemistry
Tetrahedron Elsevier
Green Chemistry Royal Society of Chemistry
Tetrahedron Letters Elsevier
Helvetica Chimica Acta Wiley-VCH
Tetrahedron: Asymmetry Elsevier
(continued)
to be read by other users via appropriate news software. An example of a

usernet group is The Higher Education Academy (in the UK) that supports
the Physical Sciences Centre which supports staff and students in Higher
Education by promoting the enhancement of the student learning experience
in chemistry, physics, astronomy and forensic science. Individuals can sign
up to receive the email mailing list, RSS feeds to receive regular updates on
news, blogs and periodical publications and Twitter.
Key Point Remember that the information from Internet

newsgroups and similar websites may be unedited and may
represent the personal opinion of the author of the article.

Internet resources
Table 44.2 Selected websites Chemical databases

Type Name Website A wide range of chemical databases are available on the Internet. Unfortunately
some of them require a subscription to access them; however, your university
Professional Royal Society http://www maybe already be a subscriber. You will need to ask your library staff to find
bodies of Chemistry .rsc.org/
out. Specific examples of some on these databases are now illustrated.
American http://www
hemical
C .acs.org/ Periodic Table
Society
A good place to start for a chemistry student is finding information about the
Non- International http://www elements in the Periodic Table. Various organisations are offering interactive
governmental Union of Pure .iupac.org/
body and Applied
Periodic Tables that include blogs, videos and interactive screens for each ele-
Chemistry ment. Examples of selected on-line Periodic Table suppliers are shown below:
(IUPAC)
●● Royal Society of Chemistry (http://www.rsc.org/periodic-table)
Government National http://www
bodies Institute of .nist.gov/ ●● Dynamic Periodic Table (http://www.ptable.com/)
Standards and
Technology ●● The CRC Press Periodic Table (http://www.chemnetbase.com/periodictable/)
(NIST)
●● Los Alomos National Laboratory (http://periodic.lanl/gov/)
aboratory
L http://www
of the .lgcgroup ●● The Sheffield Chemdex includes WebElements™ (https://www.webelements
G overnment .com/ .com/)
Chemist (LGC)
●● Element Collection, Inc. (http://www.periodictable.com/)
One such resource is WebElements (http://www.webelements.com/) which pro-
vides key information on all elements in the Periodic Table. This database is
searchable in terms of a range of properties of the elements including:
●● The essentials (includes names, symbol, atomic number and atomic weight;
block, period and group in periodic table; description; standard state; registry
number; and isolation)
●● History (meaning of name; discovery; and history of the element)
●● Uses
●● Geology (abundance of elements in the universe; the sun; meteorites; Earth’s
crust; oceans; and streams)
●● Biology (abundance in humans; biological role; and health hazards)
●● Compounds (halides, oxides, sulphides, hydrides and complexes; lattice
energies; and reduction potentials)
●● Electronegativity (Pauling; Sanderson; Allred Rochow; Mulliken-Jaffe; and
Allen)
●● Bond enthalpies (of diatomic species)
●● Lattice energies
●● Physical properties (includes boiling point; melting point; density; molar
volume; thermal conductivity; and electrical resistivity; bulk modulus; crit-
ical temperature; superconductivity temperature; hardness (mineralogical,
Brinell and Vickers); linear expansion coefficient; Poisson’s ratio; reflectiv-
ity; refractive index; rigidity modulus; Young’s modulus; velocity of sound)
●● Images (i.e. photographs of the elements)
●● Reactions (reactions of element with air; water; halogens; acids; and bases)
●● Crystal structure

Internet resources
●● Thermochemistry (enthalpies of atomisation, fusion, and vaporisation; ther-

modynamic properties)
●● Electron shell properties (electronic configuration; term symbol; electron
affinity; ionisation energies; and atomic spectra)
●● Atom sizes (atomic radius; Shannon and Pauling ionic radii; covalent radius;
metallic radius; element bond length; and Van der Waals radius)
●● Atomic orbital properties (effective nuclear charge; electron binding ener-
gies; and valence orbital radii maxima)
●● Isotopes (isotope abundances; radioactive isotopes; isotope masses; nuclear
spins; and nuclear magnetic moment)
●● NMR properties (frequencies; isotopes; magnetogyric ratios; quadrupole
moments; receptivities; and relative sensitivities)
Royal Society of Chemistry (RSC) databases

The RSC provides access to three databases (Fig. 44.1) and as well as litera-
ture updating services (Fig. 44.2) and the National Chemical Database Service
(http://cds.rsc.org/) (Fig. 44.3). The three databases are:
●● MarinLit: A comprehensive database of the marine natural products litera-
ture, covering new and revised structures, synthesis, ecology and biological
activities. Features flexible searching and powerful dereplication tools based
on predicted and experimental data
●● The Merck Index Online (https://www.rsc.org/merck-index). The Merck
Index (Fig. 44.4) has been regarded as the most authoritative and reliable
Fig. 44.1 Royal Society of Chemistry databases.

Internet resources
Fig. 44.2 Royal Society of Chemistry: Literature updating services.
Fig. 44.3 The National Chemical Database Service.

Internet resources
Fig. 44.4 The Merck Index Online.
source of information on chemicals, drugs and biologicals for over 120 years.
The Merck Index Online offers the same highly authoritative information as
the print edition in a convenient and easily searchable full text database. It
contains over 11 500 monographs – including historic records not available
in the print edition. Users can perform simple searches on a number of dif-
ferent properties and fields, construct complex multi-parameter searches, and
search by chemical structure (see Table 44.3). An example of a simple search
for the named reaction: Diels-Alder is shown in Fig. 44.5.
●● ChemSpider (http://www.chemspider.com/). ChemSpider is a free chemical
structure database providing fast text and structure search access to over
43 million structures from hundreds of data sources (Fig. 44.6). In addition,
Table 44.3 Merck Index Online
Topic Content
Substance ●● Names, synonyms and tradenames

monographs
●● Chemical structures
●● Physical properties (e.g. mp, bp, density)
●● Applications and usage
●● Bioactivity data
●● Literature references
Named ●● Searchable collection of over 500 organic named reactions
reactions
●● Overview of each reaction with an idealised scheme and key
references

Internet resources
Topic Content
Reference ●● Common abbreviations

tables
●● Conversions
●● Glossaries
Coverage ●● Human and veterinary drugs
●● Biotech drugs and monoclonal antibodies
●● Substances used for medical imaging
●● Biologicals and natural products
●● Plant and herbal medicines
●● Nutraceuticals and cosmeceuticals
●● Laboratory reagents and catalysts
●● Dyes, colour and indicators
●● Environmentally significant substances
●● Food additives and nutritional supplements
●● Flavours and fragrances
●● Agricultural chemicals, pesticides and herbicides
●● Industrial and specialty chemicals
ChemSpider: Synthetic pages (http://cssp.chemspider.com/) (Fig.44.7) is

a free database of practical procedures for research workers in synthetic
chemistry (organic, organometallic and inorganic), written by chemists for
chemists. An example is shown in Table 44.4.
Fig. 44.5 An example of a search for a named reaction: Diels-Alder.

Internet resources
Fig. 44.6 ChemSpider webpage.
Fig. 44.7 ChemSpider: Synthetic pages.

Internet resources
Table 44.4 An example from Chemspider: Synthetic pages illustrating the typical information that is available
Aldol condensation of benzaldehyde and acetone; trans-dibenzalacetone, (lE,4E)-1,5 Diphenyl-1,4-pentadien-3-one, DBA
SyntheticPage 771
DOI: 10.1039/SP771
Submitted Dec 19, 2014, published Dec 21, 2014
Matthew McBride (mcbridemj@gmail.com), Jean-Claude Bradley
A contribution from UsefulChem
Chemicals Used
Benzaldehyde (Sigma Aldrich, 98%)
Acetone (Sigma Aldrich, 99%)
Sodium Hydroxide (NaOH)
Ethanol
Distilled Water
Procedure
A solution of NaOH (0.902 g, 22.55 mmol, 1.6 eq) in distilled water (40 mL) at room temperature was added to a stirred solution
of benzaldehyde (4.361 g, 41.13 mmol, 2.9 eq) in ethanol (40 mL). Acetone (0.833 g, 14.36 mmol, 1.0 eq) was added to the reaction
mixture. The reaction mixture was then stirred at room temperature for 30 minutes. The precipitated product was recovered by
suction filtration, washed 1x with 2–3 mL of 1:1 ethanol/water, dried over suction for 25 minutes, and recovered (3.044 g, 90.6%) as
yellow crystals (mp 103-104°C).
Author’s Comments
●● This procedure produced pure trans-dibenzalacetone (verified by NMR spectroscopy) without having to recrystallise. However,
the measured melting point of the product (103–104°C) is less than the literature value (110–112°C). If needed, the product may be
recrystallise from ethanol.
●● The benzaldehyde must be dissolved in the ethanol prior to addition to the water solution.
Data
1
H NMR Spectrum (500 MHz, CDCl3): d 7.72 (d, J = 16.3 Hz, 2H), d 7.58 (m, 4H), d 7.38 (m, 6H), d 7.06 (d, J = 15.5 Hz, 2H)
Lead Reference
B. L. Hawbecker, D. W. Kurtz, T. D. Putnam, P. A. Ahlers, G. D. Gerber, Journal of Chemical Education 1978 55(8), 540
http:dx.doi.org/10.1021/ed055p540
Other References
M. J. McBride, 1. C. Bradley. UsefulChem, Experiment 284. http://usefulchem.wikispaces.com/EXP284
Supplementary Information: 1H NMR Spectrum
Keywords: Aldol Condensation

Internet resources
Other chemical databases

The CHEMnetBASE (http://www.chemnetbase.com/) offers a listing to some
of the most frequently used chemical databases. Specifically, the Handbook
of Chemistry and Physics (www.hbcponline.com) (Fig. 44.8) has traditionally
been available in hardback copy for nearly 100 years (the 3rd edition of this
book is citing as a reference the 96th edition of the CRC Handbook of Chem-
istry and Physics). The online version allows a range of menu-driven pages to
allow:
●● Searching by various fields – physical property, molecular formula, etc.
●● Cross table searching – displays everything about a specific compound
●● Chemical structure searching – use the free Applet to draw a desired
structure
●● Interactive tables – customise the data in spreadsheet format
The contents of the 96th on-line edition (2015-16) are shown in Table 44.5.
MassBank (http://www.massbank.jp): MassBank is the first public repos-
itory of mass spectral data (Fig. 44.9) for sharing among scientific research
community. MassBank data are useful for the chemical identification and
structure elucidation of chemical compounds detected by mass spectrometry.
By searching for a named compound, e.g. chlorobenzene, the mass spectrum
for that compound can be identified (Fig. 44.10); peaks at 77 m/z indicate a
monosubstituted benzene ring while the 3:1 ratio of the peaks at 112 and 114
would indicate the presence of chlorine.
Fig. 44.8 Handbook of Chemistry and Physics Online.

Internet resources
Table 44.5 The content of the 96th online edition (2015-16) of the CRC Handbook of Chemistry & Physics
Introduction ●● Section 4: Properties of the Elements and Inorganic

Section 1: Basic Constants, Units and Conversion Factors Compounds
●● CODATA Recommended Values of the Fundamental Physi- ●● The Elements

cal Constants: 2010 ●● Physical Constants of Inorganic Compounds
●● Standard Atomic Weights (2013) ●● Physical Properties of the Rare Earth Metals
●● Atomic Masses and Abundances ●● Melting, Boiling, Triple and Critical Point Temperatures of
●● Electron Configuration and Ionisation Energy of Neutral the Elements
Atoms in the Ground State ●● Heat Capacity of the Elements at 25°C
●● International Temperature Scale of 1990 (ITS-90) ●● Vapour Pressure of the Metallic Elements – Equations
●● Conversion of Temperatures from the 1948 and 1968 ●● Vapour Pressure of the Metallic Elements – Data
Scales to ITS-90
●● Density of Molten Elements and Representstive Salts
●● International System of Units (SI)
●● Magnetic Susceptibility of the Elements and Inorganic
●● Units for Magnetic Properties Compounds
●● Conversion Factors ●● Index of Refraction of Inorganic Liquids
●● Conversion of Temperatures ●● Physical and Optical Properties of Minerals
●● Conversion Factors for Energy Units ●● Crystallographic Data on Minerals
●● Conversion Factors for Pressure Units
Section 5: Thermochemistry, Electrochemistry and Solution
●● Conversion Factors for Thermal Conductivity Units Chemistry
●● Conversion Factors for Electrical Resistivity Units ●● CODATA Key Values for Thermodynamics
●● Conversion Formulas for Concentration of Solutions ●● Standard Thermodynamic Properties of Chemical
Substances
●● Descriptive Terms for Solubility
●● Thermodynamic Properties as a Function of Temperature
●● Conversion Factors for Chemical Kinetics
●● Thermodynamic Properties of Aqueous Ions
●● Conversion Factors for Ionising Radiation
●● Heat of Combustion
●● Values of the Gas Constant in Different Unit Systems
●● Energy Content of Fuels
●● Periodic Table of the Elements
●● Ionisation Constant of Water
Section 2: Symbols, Terminology and Nomenclature
●● Ionisation Constant of Normal and Heavy Water
●● Symbols and Terminology for Physical and Chemical
Quantities ●● Electrical Conductivity of Water
●● Expression of Uncertainty of Measurements ●● Electrical Conductivity of Aqueous Solutions
●● Nomenclature of Chemical Compounds ●● Standard KCI Solutions for Calibrating Conductivity Cells
●● Nomenclature for Inorganic Ions and Ligands ●● Molar Conductivity of Aqueous HF, HCI, HBr and HI
●● Organic Substituent Groups and Ring Systems ●● Equivalent Conductivity of Electrolytes in Aqueous
Solution
●● Representation of Chemical Structures with the IUPAC
International Chemical Identifier (InChl) ●● Ionic Conductivity and Diffusion at Infinite Dilution
●● Scientific Abbreviations, Acronyms and Symbols ●● Electrochemical Series
●● Greek, Russian and Hebrew Alphabets ●● Reduction and Oxidation Potentials for Certain Ion Radicals
●● Definitions of Scientific Terms ●● Dissociation Constants of Inorganic Acids and Bases
●● Thermodynamic Functions and Relations ●● Dissociation Constants of Organic Acids and Bases
●● Nobel Laureates in Chemistry and Physics ●● Activity Coefficients of Acids, Bases and Salts
Section 3: Physical Constants of Organic Compounds ●● Mean Activity Coefficients of Electrolytes as a Function of
Concentration
●● Physical Constants of Organic Compounds
●● Enthalpy of Dilution of Acids
●● Diamagnetic Susceptibility of Selected Organic
Compounds ●● Enthalpy of Solution of Electrolytes
(continued)

Internet resources
Table 44.5 Continued
●● Enthalpy of Hydration of Gases ●● Recommended Data for Vapour-Pressure Calibration

●● pH Scale for Aqueous Solutions ●● Enthalpy of Vapourization
●● Practical pH Measurements on Natural Waters ●● Enthalpy of Fusion
●● Buffer Solutions Giving Round Values of pH at 25°C ●● Compressibility and Expansion Coefficients of Liquids
●● Concentrative Properties of Aqueous Solutions: Density, ●● Temperature and Pressure Dependence of Liquid Density
Refractive Index, Freezing Point Depression, and Viscosity
●● Volumetric Properties of Aqueous Sodium Chloride
●● Solubility of Selected Gases in Water Solutions
●● Solubility of Carbon Dioxide in Water at Various Tempera- ●● Properties of Cryogenic Fluids
tures and Pressures
●● Properties of Liquid Helium
●● Aqueous Solubility and Henry’s Law Constants of Organic
Compounds ●● Properties of Refrigerants
●● Aqueous Solubility of Inorganic Compounds at Various ●● Properties of Gas Clathrate Hydrates

Temperatures ●● Ionic Liquids
●● Octanol-Water Partition Coefficients
●● Density and Specific Volume of Mercury
●● Solubility Product Constants
●● Thermal Properties of Mercury
●● Solubility of Common Salts at Ambient Temperatures
●● Melting Curve of Mercury
●● Solubility of Hydrocarbons in Seawater
●● Vapor Pressure of Mercury
●● Solubility of Organic Compounds in Pressurised Hot Water
●● Surface Tension of Common Liquids
●● Solubility Chart
●● Surface Tension of Aqueous Mixtures
Section 6: Fluid Properties ●● Permittivity (Dielectric Constant) of Liquids
●● Thermophysical Properties of Water and Steam ●● Permittivity (Dielectric Constant) of Gases
●● Vapour Pressure and other Saturation Properties of Water ●● Azeotropic Data for Binary Mixtures
●● Standard Density of Water ●● Viscosity of Gases
●● Fixed Point Properties of H2O and D2O ●● Viscosity of Liquids
●● Properties of Saturated Liquid D2O ●● Viscosity of Carbon Dioxide along the Saturation Line
●● Properties of Ice and Supercooled Water ●● Viscosity and Density of Aqueous Hydroxide Solutions
●● Vapour Pressure of Ice ●● Viscosity of Liquid Metals
●● Melting Point of Ice as a Function of Pressure
●● Thermal Conductivity of Gases
●● Permittivity (Dielectric Constant) of Water at Various
●● Thermal Conductivity of Liquids
Frequencies
●● Diffusion in Gases
●● Thermophysical Properties of Air
●● Diffusion of Gases in Water
●● Thermophysical Properties of Fluids
●● Thermophysical Properties of Selected Fluids at ●● Diffusion Coefficients in Liquids at Infinite Dilution
Saturation Section 7: Biochemistry
●● Virial Coefficients of Selected Gases ●● Properties of Amino Acids
●● Van der Waals Constants for Gases ●● Structures of Common Amino Acids
●● Mean Free Path and Related Properties of Gases ●● Properties of Purine and Pyrimidine Bases
●● Influence of Pressure on Freezing Points
●● The Genetic Code
●● Critical Constants of Organic Compounds
●● Properties of Fatty Acids and their Methyl Esters
●● Critical Constants of Inorganic Compounds
●● Properties of Fatty Acid Methyl and Ethyl Esters Related to
●● Sublimation Pressure of Solids Biofuels
●● Vapour Pressure ●● Composition and Properties of Common Oils and Fats
●● Vapour Pressure of Fluids at Tevmperatures below 300 K ●● Carbohydrate Names and Symbols
●● Vapour Pressure of Saturated Salt Solutions

Internet resources
●● Standard Transformed Gibbs Energy of Formation for Bio- ●● Proton NMR Absorption of Major Chemical Families
chemical Reactants
●● Proton NMR Correlation Chart for Major Organic Func-
●● Apparent Equilibrium Constants for Enzyme-Catalysed tional Groups
Reactions
●● Proton NMR Shifts of Common Organic Solvents
●● Thermodynamic Quantities for the Ionisation Reactions of
●● 13C NMR Absorptions of Major Functional Groups
Buffers in Water
●● 13C NMR Chemical Shifts of Common Organic Solvents
●● Biological Buffers
●● 15N NMR Chemical Shifts of Major Chemical Families
●● Typical pH Values of Biological Materials and Foods
●● Natural Abundance of Important Isotopes
●● Structure and Functions of Some Common Drugs
●● Common Mass Spectral Fragmentation Patterns of
●● Chemical Constituents of Human Blood
Organic Compound Families
●● Chemical Composition of the Human Body
●● Common Mass Spectral Fragments Lost
●● Nutrient Values of Foods
●● Major Reference Masses in the Spectrum of Hepta-
Section 8: Analytical Chemistry cosafluorotributylamine (Perfluorotributylamine)
●● Abbreviations and Symbols Used in Analytical Chemistry ●● Mass Spectral Peaks of Common Organic Solvents
●● Basic Instrumental Techniques of Analytical Chemistry ●● Common Spurious Signals Observed in Mass
Spectrometers
●● Analytical Standardization and Calibration
●● Chlorine-Bromine Combination Isotope Intensities
●● Figures of Merit
●● Reduction of Weighings in Air to Vacuo
●● Mass- and Volume-Based Concentration Units
●● Standards for Laboratory Weights
●● Detection of Outliers in Measurements
●● Indicators for Acids and Bases
●● Properties of Carrier Gases for Gas Chromatography
●● Preparation of Special Analytical Reagents
●● Common Symbols Used in Gas and Liquid Chromato-
graphic Schematic Diagrams ●● Organic Analytical Reagents for the Determination of Inor-
ganic Ions
●● Stationary Phases for Porous Layer Open Tubular
Columns ●● Precipitation of Sulfides
●● Coolants for Cryotrapping ●● pH Range for Precipitation of Metal Hydroxides and

Oxides
●● Properties of Common Cross-Linked Silicone Stationary
Phases Section 9: Molecular Structure and Spectroscopy
●● Detectors for Gas Chromatography ●● Bond Lengths in Crystalline Organic Compounds
●● Varieties of Hyphenated Gas Chromatography with Mass ●● Bond Lengths in Organometallic Compounds
Spectrometry
●● Structure of Free Molecules in the Gas Phase
●● Solid-Phase Microextraction Sorbents
●● Characteristic Bond Lengths in Free Molecules
●● Gas Chromatographic Retention Indices
●● Atomic Radii of the Elements
●● Eluotropic Values of Solvents on Octadecylsilane and
●● Dipole Moments
Octytsilane
●● Hindered Internal Rotation
●● Instability of HPLC Solvents
●● Bond Dissociation Energies
●● Detectors for Liquid Chromatography
●● Electronegativity
●● Solvents for Ultraviolet Spectrophotometry
●● Force Constants for Bond Stretching
●● Correlation Table for Ultraviolet Active Functionalities
●● Fundamental Vibrational Frequencies of Small Molecules
●● Wavelength-Wavenumber Conversion Table
●● Spectroscopic Constants of Diatomic Molecules
●● Middle-Range Infrared Absorption Correlation Charts
Section 10: Atomic, Molecular and Optical Physics
●● Common Spurious Infrared Absorption Bands
●● Line Spectra of the Elements
●● Nuclear Spins, Moments and Other Data Related to NMR
Spectroscopy ●● Atomic Transition Probabilities
●● Properties of Important NMR Nuclei ●● Electron Affinities
(continued)

Internet resources
●● Proton Affinities ●● Permittivity (Dielectric Constant) of Inorganic Solids

●● Atomic and Molecular Polarisabilities ●● Curie Temperature of Selected Ferroelectric Crystals
●● Ionisation Energies of Atoms and Atomic Ions ●● Properties of Antiferroelectric Crystals
●● Ionisation Energies of Gas-Phase Molecules ●● Dielectric Constants of Glasses
●● X-Ray Atomic Energy Levels ●● Properties of Superconductors
●● Electron Binding Energies of the Elements ●● High Temperature Superconductors
●● Natural Width of X-Ray Lines ●● Organic Superconductors
●● Photon Attenuation Coefficients ●● Properties of Semiconductors
●● Classification of Electromagnetic Radiation ●● Selected Properties of Semiconductor Solid Solutions
●● Sensitivity of the Human Eye to Light of Different ●● Properties of Organic Semiconductors
Wavelengths
●● Diffusion Data for Semiconductors
●● Blackbody Radiation
●● Properties of Magnetic Materials
●● Characteristics of Infrared Detectors
●● Organic Magnets
●● Index of Refraction of Inorganic Crystals
●● Electron inelastic Mean Free Paths
●● Refractive Index and Transmittance of Representative
●● Electron Stopping Powers
Glasses
●● Electron Work Function of the Elements
●● Index of Refraction of Water
●● Secondary Electron Emission
●● Index of Refraction of Liquids for Calibration Purposes
●● Optical Properties of Selected Elements
●● Index of Refraction of Air
●● Optical Properties of Selected Inorganic and Organic Solids
●● Index of Refraction of Gases
●● Elasto-optic, Electro-optic, and Magneto-optic Constants
●● Characteristics of Laser Sources
●● Nonlinear Optical Constants
●● Infrared Laser Frequencies
●● Phase Diagrams
●● Infrared and Far-Infrared Absorption Frequency Standards
●● Properties of Selected Materials at Cryogenic
Section 11: Nuclear and Particle Physics Temperatures
●● Summary Tables of Particle Properties ●● Heat Capacity of Selected Solids
●● Table of the Isotopes ●● Thermal and Physical Properties of Pure Metals
●● Neutron Scattering and Absorption Properties ●● Thermophysical Properties of Stainless Steel 310
●● Cosmic Radiation ●● Thermal Conductivity of Metals and Semiconductors as a
Section 12: Properties of Solids Function of Temperature
●● Techniques for Materials Characterisation ●● Thermal Conductivity of Alloys as a Function of

Temperature
●● Symmetry of Crystals
●● Thermal Conductivity of Crystalline Dielectrics
●● Ionic Radii in Crystals
●● Thermal Conductivity of Ceramics and Other Insulating
●● Polarisability of Atoms and ions in Solids Materials
●● Crystal Structures and Lattice Parameters of Allotropes of ●● Thermal Conductivity of Glasses
the Elements
●● Thermoelectric Properties of Metals and Semiconductors
●● Phase Transitions in the 5olid Elements at Atmospheric
Pressure ●● Fermi Energy and Related Properties of Metals
●● Lattice Energies ●● Properties of Commercial Metals and Alloys
●● The Madelung Constant and Crystal Lattice Energy ●● Hardness of Minerals and Ceramics
●● Elastic Constants of Single Crystals Section 13: Polymer Properties
●● Electrical Resistivity of Pure Metals ●● Abbreviations Used in Polymer Science and Technology
●● Electrical Resistivity of Selected Alloys ●● Physical Properties of Selected Polymers
●● Electrical Resistivity of Graphite Materials

Internet resources
●● Nomenclature for Organic Polymers Section 15: Practical Laboratory Data

●● Solvents for Common Polymers ●● Standard ITS-90 Thermocouple Tables
●● Glass Transition Temperature for Selected Polymers ●● Reference Points on the ITS-90 Temperature Scale
●● Dielectric Constant of Selected Polymers ●● Relative Sensitivity of Bayard-Alpert Ionisation Gauges to
Various Gases
●● Second Virial Coefficients of Polymer Solutions
●● Laboratory Solvents and other Liquid Reagents
●● Pressure-Volume-Temperature Relationship for Polymer
Melts ●● Miscibility of Organic Solvents
●● Upper Critical (UCST) and Lower Critical (LCST) Solution ●● Density of Solvents as a Function of Temperature
Temperatures of Binary Polymer Solutions
●● Dependence of Boiling Point on Pressure
●● Vapour Pressures (Solvent Activities) for Binary Polymer
●● Ebullioscopic Constants for Calculation of Boiling Point
Solutions
Elevation
●● Specific Enthalpies of Solution of Polymers and
●● Cryoscopic Constants for Calculation of Freezing Point
Copolymers
Depression
●● Solubility Parameters of Selected Polymers
●● Freezing Point Lowering by Electrolytes in Aqueous Solution
Section 14: Geophysics, Astronomy and Acoustics
●● Correction of Barometer Readings to 0° C Temperature
●● Astronomical Constants ●● Determination of Relative Humidity from Dew Point
●● Properties of the Solar System ●● Determination of Relative Humidity from Wet and Dry
●● Satellites of the Planets Bulb Temperatures
●● Interstellar Molecules ●● Constant Humidity Solutions
●● Mass, Dimensions and other Parameters of the Earth ●● Standard Salt Solutions for Humidity Calibration
●● Geological Time Scale ●● Low Temperature Baths for Maintaining Constant

Temperature
●● Acceleration Due to Gravity
●● Metals and Alloys with Low Melting Temperature
●● Density, Pressure and Gravity as a Function of Depth
within the Earth ●● Wire Tables
●● Ocean Pressure as a Function of Depth and Latitude ●● Standard Fittings for Compressed Gas Cylinders
●● Properties of Seawater ●● Plug and Outlet Configurations for Common Laboratory

Devices
●● Abundance of Elements In the Earth’s Crust and in the Sea
●● Characteristics of Particles and Particle Dispersoids
●● Solar Irradiance at the Earth
●● Density of Various Solids
●● U.S. Standard Atmosphere (1976)
●● Density of Sulphuric Acid
●● Geographical and Seasonal Variation in Solar Radiation
●● Density of Ethanol-Water Mixtures
●● Major World Earthquakes
●● Dielectric Strength of Insulating Materials
●● Weather-Related Scales
●● Coefficient of Friction
●● Infrared Absorption by the Earth’s Atmosphere
●● Flame Temperatures
●● Atmospheric Concentration of Carbon Dioxide, 1958–2014
●● Allocation of Frequencies in the Radio Spectrum
●● Global Temperature Trend, 1880–2014
Section 16: Health and Safety Information
●● Global Warming Potential of Greenhouse Gases
●● Abbreviations Used in the Assessment and Presentation
●● Atmospheric Electricity
of Laboratory Hazards
●● Speed of Sound in Various Media
●● Incompatible Chemicals
●● Attenuation and Speed of Sound in Air as a Function of
●● Explosion (Shock) Hazards
Humidity and Frequency
●● Water-Reactive Chemicals
●● Speed of Sound in Dry Air
●● Testing Requirements for Peroxidisable Compounds
●● Musical Scales
●● Tests for the Presence of Peroxides
●● Characteristics of Human Hearing
(continued)

Internet resources
●● Pyrophoric Compounds – Compounds That Are Reactive Binomial Series

with Air
Reversion of Series
●● Flammability Hazards of Common Solvents
Taylor Series
●● Selection of Laboratory Gloves
Exponential Series
●● Selection of Protective Laboratory Garments
Logarithmic Series
●● Selection of Respirator Cartridges and Filters
Trigonometric Series
●● Materials Compatible with and Resistant to 72%
●● Transforms
Perchloric Acid
Fourier Transforms
●● Protective Clothing Levels
Table of Fourier Cosine Transforms
●● Chemical Fume Hoods and Biological Safety Cabinets
Table of Finite Cosine Transforms
●● Gas Cylinder Safety and Stamped Markings
Table of Fourier Sine Transforms
●● Flammability of Chemical Substances
Table of Finite Sine Transforms
●● Threshold Limits for Airborne Contaminants
Table of Fourier Transforms
●● Laser Hazards in the Laboratory
Table of Functional Relations for Fourier Transforms
●● General Characteristics of Ionising Radiation for the Pur-
pose of Practical Application of Radiation Protection Table of Multidimensional Fourier Transforms
●● Radiation Safety Units Table of Laplace Transforms
●● Relative Dose Ranges from Ionising Radiation Table of Functional Relations for Laplace Transforms
●● Annual Limits on Intakes of Radionuclides ●● Special Functions
●● Chemical Carcinogens Orthogonal Polynomials
Section 17: Mathematical Tables Tables of Orthogonal Polynomials
●● Constants Bessel Functions
Mathematical Constants Factorial Function
Decimal Equivalents of Fractions (inches to mm) Gamma Function
Exponential and Hyperbolic Functions and their Common Beta Function

Logarithms Error Function
Trigonometric Functions to Four Decimal Places ●● Probability
●● Algebra Normal Probability Function
Quadratic Formula Confidence Intervals
Vector Algebra Percentage Points, Student’s t-Distribution
●● Geometry Percentage Points, Chi-Square Distribution
Geometry of the Plane, Straight Line and Sphere Percentage Points, F-Distribution
Geometry of Curves in Space ●● Physics Related
●● Trigonometry Clebsch-Gordan Coefficients
Trigonometric Functions in Terms of One Another Moment of Inertiol for Different Shapes
Hyperbolic Functions in Terms of One Another ●● References
●● Calculus Section 18: Sources of Physical and Chemical Data
Differentiation
●● Sources of Physical and Chemical Data
Orthogonal Coordinate Systems
Section 19: Tables from Older Editions
Integration
●● Tables Relocated or Removed from CRC Handbook of
Differential Equations Chemistry and Physics, 71st through 96th Editions
●● Series ●● Allowable Carrying Capacities of Conductors
Fourier Series

Internet resources
●● Ammine Complexes of Metals ●● Platinum Wire

●● Aluminium Wire Table ●● Practical pH Measurements on Natural Waters
●● Brazing Filler Metals (Solders) ●● Properties of Carbohydrates
●● Chemical Composition of Rocks ●● Properties of Sulphuric Acid
●● Chemical Kinetic Data for Stratospheric Modelling ●● Properties of Tungsten
●● Chemical Kinetic Data for Combustion Modelling ●● Protection against Ionising Radiation
●● Classification of Comparative Life Hazards of Gases and ●● Radiative Transition Probabilities for X-Ray Lines
Vapours
●● Radioactive Tracer Diffusion Data for Pure Metals
●● Constants for Satellite Geodesy
●● Recommended Daily Dietary Allowances
●● Cross-Section and Mass of Wires
●● Reduction of Barometer to Sea Level
●● Density and Composition of Fuming Sulphuric Acid
●● Refractory Materials
●● Diamagnetic Susceptibility Data on Organosilicon
Compounds ●● Solvents for Liquid Chromatography
●● Diffusivities of Metallic Solutes in Molten Metals ●● Spark-Gap Voltages
●● Diffusivities of Metallic Tracers in Mercury ●● Specific Heat and Enthalpy of Some Solids at Low
Temperature
●● Dissociation Constants of Acids in Water at Various
Temperatures ●● Spectral Emissivity
●● Dissociation Constants of Aqueous Ammonia from 0 to ●● Spectral Emissivity of Oxides

50°C ●● Standard Solutions of Acids, Bases and Salts
●● Efficacies and Other Characteristics of llluminants ●● Standard Solutions of Oxidation and Reduction Reagents
●● Efficiency of Drying Agents
●● Standard Test Sieves and Mesh Size Conversion
●● Emergent Stem Corrections for Liquid-in-Glass
●● Standard Types of Stainless and Heat Resisting Steels
Thermometers
●● Steroid Hormones and Other Steroidal Synthetics
●● Emissivity of Total Radiation for Various Materials
●● Surface Tension of Liquid Elements
●● Emissivity of Tungsten
●● Temperature Correction for Glass Volumetric Apparatus
●● Fats and Oils
●● Temperature Correction for Volumetric Solutions
●● Flame and Bead Tests
●● Temperature Correction, Glass Scale
●● Fluorescent Indicators
●● The Limits of Superheat of Pure Liquids
●● Handling and Disposal of Chemicals in Laboratories
●● Thermal Conductivity of Rocks
●● Infrared Correlation Charts
●● Introduction to X-Ray Cross Sections ●● Total Monthly Solar Radiation in a Cloudless Sky
●● Ion Exchange Resins ●● Transmission of Corning Colored Filters
●● Lattice Constants for Cubic Crystals ●● Transmission of Wratten Filters
●● Lattice Spacing of Common Analysing Crystals ●● Ultraviolet Spectra of Common Liquids
●● Lowering of Vapour Pressure by Salts in Aqueous Solution ●● Units, Symbols and Equations for Radiometric and
Photometric Quantities
●● Magnetic Rotatory Power
●● Values for the Langevin Function
●● Mean Temperatures in the United States, 1900–1992
●● Van der Waals Constants for Gases
●● Nomograph and Table for Doppler Linewidths
●● Volume of One Gram of Water
●● Organic Analytical Reagents for the Determination of Inor-
ganic Compounds ●● X-Ray Wavelengths
●● Oxygen Solubility in Aqueous Electrolyte Solutions Section 20: Index

●● Physical Constants of Clear Fused Quartz ●● Index

Internet resources
Crystallography Open Database (http://www.crystallography.net/).

An open-access collection of crystal structures of organic, inorganic,
metal-organic compounds and minerals, excluding biopolymers (Fig. 44.11).
It can be searched as follows: for example, select ‘Chemical Communica-
tions’ (Fig. 44.12 (a)). Then, by selecting one of the entries (Fig. 44.12 (b))
i.e. 7118949, click on the ‘COD ID’ (Fig. 44.12 (c), then select ‘structure
parameters’ (Fig. 44.12(d)).
‘MassBank: A public repository for sharing mass spectral data for life sciences’,
H. Horai, M. Arita, S. Kanaya, V. Nihei, T. Ikeda, K. Suwa. Y. Ojima, K. Tanaka,
S. Tanaka, K. Aoshima, Y. Oda, Y. Kakazu, M. Kusano, T. Tohge, F. Matsuda,
Y. Sawada, M. Yokota Hirai, H. Nakanishi, K. Ikeda, N. Akimoto, T. Maoka,
H. Takahashi, T. Ara, N. Sakurai, H. Suzuki, D. Shibata, S. Neumann, T. Iida,
K. Tanaka, K. Funatsu, F. Matsuura, T. Soga, R. Taguchi, K. Saito and
T. Nishioka, J. Mass Spectrom., 45, 703–714 (2010) .
Fig. 44.9 MassBank: contains chemical identification and structure elucidation of chemical compounds detected by
mass spectrometry.

Internet resources
Fig. 44.10 Mass spectral search of MassBank for chlorobenzene.
Fig. 44.11 The Crystallography Open Database.

Internet resources
(a)
Fig. 44.12 Use of Crystallography Open Database (COD) database.
(b)
Fig. 44.12 Continued

Internet resources
(c)
(d)
Fig. 44.12 Continued

Internet resources
Study exercises
44.1 Investigate the potential of the internet for locat- (a) Who is the publisher of this journal?
ing information in the Periodic Table. Using the (b) What is the journal’s Impact Factor?
web, answer the following questions: (c) Who is the chair of the editorial board?
(d) What type of articles does the journal publish?
(a) What is the chemical symbol for
praseodymium? 44.3 Search the MassBank (http://www.massbank
(b) What is its atomic number? .jp/) database for the spectrum for bromobenzene.
(c) What group of the Periodic Table is the ele-
(a) What does the 77 m/z ion represent?
ment located in?
(b) What features are characteristic of bromine?
(d) What is praseodymium’s electronic
(c) How many stable isotopes does bromine have?
configuration.
(d) How would you confirm that it is
44.2 Search for the journal: Chemical Society Reviews bromobenzene?

45 Using spreadsheets
The spreadsheet is one of the most powerful and flexible computer applica-
Definitions tions. It can be described as the electronic equivalent of a paper-based longhand
calculation, where the sums are carried out automatically. Spreadsheets provide
Macro – a sequence of user-defined
a dynamic method of storing, manipulating and analysing data sets. Advantages
instructions carried out by the spread-
of spreadsheets include:
sheet, allowing complex repeated tasks
to be ‘automated’. ●● Ease and convenience – especially when complex calculations are repeated
Office suite – a package of complemen- on different sets of data.
tary and integrated programs such as
a word processor, a spreadsheet and a ●● Accuracy – providing the entry data and cell formulae are correct, the result
database. will be free of calculation errors.
Spreadsheet – a display of a grid of ●● Improved presentation – data can be produced in graphical or tabular form
cells into which numbers, text or formu- to a very high quality.
lae can be typed to form a worksheet.
Each cell is uniquely identifiable by its ●● Integration with other programs – graphs and tables can be exported to
column and row number combination other compatible programs, such as a word processor in the same office suite.
(i.e. its 2D coordinates) and can contain
a formula which makes it possible for an ●● Useful tools – advanced features include hypothesis-testing statistics, data-
entry to one cell to alter the contents of base features and macros.
one or more other cells.
Spreadsheets can be used to:
Template – a pre-designed spreadsheet
without data but including all formulae ●● store and manipulate raw data by removing the drudgery of repeated cal-
necessary for (repeated) data analysis. culations, allowing easy transformation of data and calculation of statistics;
●● display your data Printouts can be used in practical and project reports;
●● carry out statistical analysis using in built procedures or by allowing con-
Example (Spreadsheets) struction of formulae for specific tasks;
●● Excel (Microsoft)
●● model ‘what if’ situations where the consequences of changes in data can
●● Numbers (Apple)
be seen and evaluated.
Note: This chapter uses Microsoft
Excel for illustrative purposes. A typical spreadsheet (Fig. 45.1) is divided into rows (identified by num-
bers) and columns (identified by alphabetic characters). Each individual com-
bination of column and row forms a cell which can contain either a data item,
a formula or a piece of text. Formulae can include scientific and/or statistical
functions and/or a reference to other cells or groups of cells (often called a
range). Complex systems of data input and analysis can be constructed. The
analysis, in part or complete, can be printed out. New data can be added at
any time and the sheet will recalculate automatically. The power a spreadsheet
offers is directly related to your ability to create arrays of formulae (models)
that are accurate and templates that are easy to use.
Data output from analytical instruments Data entry

– many devices provide output in
spreadsheet-compatible form (e.g. a
Spreadsheets have built-in commands which allow you to control the layout
‘comma delimited’ file). Once you have
of data in the cells (see Fig. 45.2). These include number format, the number of
uploaded the information into a spread-
decimal places to be shown (the spreadsheet always calculates using eight or
sheet, you can manipulate, analyse and
more places), the cell width and the location of the entry within the cell (left,
present it according to your needs.
right or centre). An auto-entry facility assists greatly in entering large amounts
Consult instrument manuals and the
of data by moving the entry cursor either vertically or horizontally as data are
spreadsheet help function for details.
entered. Recalculation default is usually automatic so that when a new data
value is entered the entire sheet is recalculated immediately.

Using spreadsheets
Fig. 45.1 The appearance of a typical spreadsheet, showing cells, rows and columns, toolbars, etc.
Microsoft Excel 2007 product screenshot reprinted with permission from Microsoft Corporation.
(a) (b)
Fig. 45.2 Example of cell formatting options within a Microsoft Excel spreadsheet. These menus are accessed via the
Format 7 Cell option and would apply to all of a range of selected cells. (a) Use of the number formatting option to
specify that data will be presented to three decimal places (the underlying data will be held to greater accuracy). (b) Use
of the date formatting option to specify that dates will be presented in day/month/year format. (Spreadsheet dates
are stored numerically and converted to appropriate formats. This allows a period between two dates to be calculated
more easily).
Microsoft Excel Product screenshots reprinted with permission from Microsoft Corporation.

Using spreadsheets
The parts of a spreadsheet

Labels
These should be used to identify parts of the spreadsheet, for example, stat-
ing what data are contained in a particular column or indicating that a cell’s
contents represent the end point of a calculation. In Excel, it may be useful to
use the Cells 7 Format 7 Format Cells section in the Home tab to delimit
Using hidden columns – these are use-
numerical sections of your spreadsheet. Note that spreadsheet programs have
ful for storing intermediate calculations
been designed to make assumptions about the nature of data entry being made.
which you do not wish to be displayed
If the first character is a number, then the entry is treated as numerical data; if it
on the screen or printout.
is a letter, then it is treated as a text entry; and if it is a specific symbol (‘ = ’ in
Microsoft Excel), then what follows is a formula. If you wish to enter text that
starts with a number, then you must type a designated character to show this (a
single quote mark in Microsoft Excel).
Operators and brackets in spreadsheets
– the standard mathematical operators Numbers
, and * are usually replaced by / You can also enter numbers (values) in cells for use in calculations. Many pro-
and * respectively, while ^ signifies grams let you enter numbers in more than one way and you must decide which
‘to the power’ (e.g. 10 ^ 4 = 104 ). In method you prefer. The way you enter the number does not affect the way it is
complex formulae, brackets should be displayed on the screen as this is controlled by the cell format at the point of
used to separate the elements, other- entry. There are usually special ways to enter data for percentages, currency
wise the results may not be what you and scientific notation for very large and small numbers.
expect. For example, Excel will calcu-
late = A1*B1/C1- D1 differently from Formulae
(A1*B1)/(C1-D1). These are the ‘power tools’ of the spreadsheet because they do the calculations. A
cell can be referred to by its alphanumeric code, e.g. A5 (column A, row 5) and
the value contained in that cell manipulated within a formula, e.g. =(A5 + 10)
or =(A5 + B22) in another cell. Formulae can include various pre-programmed
Definition functions which can refer to a cell, so that if the value of that cell is changed, so
is the result of the formula calculation. They may also include branching options
Function – a pre-programmed code for
the transformation of values (mathemat-
through the use of logical operators (IF, TRUE, FALSE, OR, etc.).
ical or statistical functions) or selection
of text characters (string functions). Functions
A various functions may be offered only mathematical and statistical functions
will be considered here.
Example =sin(A5) is an example of a
function in Excel. If you write this in a Mathematical functions
cell, the spreadsheet will calculate the Spreadsheets offer a range of options, including trigonometrical functions, angle
sine of the number in cell A5 (assum-
functions, logarithms and random number functions. Functions are invaluable for
ing it to be an angle in radians) and
transforming sets of data rapidly and can be used in formulae required for more
write it in the cell. Different programs
may use a slightly different syntax. complex analyses. Spreadsheets work with an order of preference of the math-
ematical operators in much the same way as a standard calculator and this must
always be taken into account when operators are used in formulae. They also
require a very precise syntax – the program should warn you if you break this.
Working with empty cells – note that
these may be given the value 0 by the Statistical functions
spreadsheet for certain functions. This Modern spreadsheets incorporate many sophisticated statistical functions, and
may cause errors, e.g. by rendering a if these are not appropriate the spreadsheet can be used to carry out the calcu-
minimum value inappropriate. Also, an lations required for most of the statistical tests found in textbooks. The descrip-
‘error return’ may result for certain func- tive statistics normally available include:
tions if the cell content is zero.
●● the sum of all data present in a column, row or block;

Using spreadsheets
●● the minimum and maximum of a defined range of cells;

●● counts of cells – a useful operation if you have an unknown or variable
number of data values;
●● averages and other statistics describing location;
●● standard deviations and other statistics describing dispersion.
A useful function where you have large data sets allows you to create
Statistical calculations – make sure
frequency distributions using pre-defined class intervals.
you understand whether the functions
The hypothesis-testing statistical functions are usually reasonably pow-
you employ apply to populations or
erful (e.g. t-test, ANOVA, regression) and they often return the probability
samples (see p. 486).
(P) of obtaining the test statistic when the null hypothesis (p. 491) is true, so
there may be no need to refer to statistical tables. Again, check on the effects
of including empty cells within the statistical calculations.
Using text functions – these allow you Database functions

to manipulate text within your spread- Many spreadsheets can be used as simple databases and offer a range of func-
sheet and include functions such as tions to support this, including filtering and sorting options. The rows and
‘find and replace’ and alphabetical or columns of the spreadsheet are used as the fields and records of the database
numerical ‘sort’. 46. For many biological purposes, this form of database is perfectly adequate
and should be seriously considered before using a full-feature database product.
Copying
(a)
Cell Formula All programs provide a means of copying (replicating) formulae or cell contents
Original Original
when required and this is a very useful feature. This is usually accomplished
cell A1 =B1+C1 formula by ‘dragging’ a cell’s contents to a new range using the mouse. When copying,
A2 =B2+C2
references to cells may be either relative, changing with the row/column as they
Copied Copied
cells formulae are copied, or absolute, remaining a fixed cell reference and not changing as
A3 =B3+C3 (relative) the formulae are copied (Fig. 45.3).
A4 =B4+C4
Key Point The distinction between relative and absolute cell

(b) references is very important and must be understood; it pro-
Cell Formula
vides one of the most common forms of error when copying
Original Original formulae.
cell A1 =B1/$C$1 formula
Copied A2 =B2/$C$1 Copied

cells formulae As an example, in Excel, copying is normally relative and if you wish a
A3 =B3/$C$1 (mixed
relative and cell reference to be absolute when copied, this is done by putting a dollar ($)
A4 =B4/$C$1 absolute) sign before and after the column reference letter, e.g. $C$56.
Fig. 45.3 Illustration of relative (a) and Naming blocks

absolute (b) copying. In Excel, the $ sign
before and after the column letter makes When a group of cells (a block) is carrying out a particular function, it is often
the cell reference absolute, as shown in (b). easier to give the block a name which can then be used in all formulae referring to
that block. This powerful feature also allows the spreadsheet to be more readable.
Spreadsheet templates
A template is a pre-constructed spreadsheet containing the formulae required
for repeated data analysis. Data are added when they become available, and
results are available as soon as the last item is entered. To create a template,
the sequence of operations is:

Using spreadsheets
1. Determine what information/statistics you want to produce.

Using templates – these should contain:
●● a data input section; 2. Identify the variables you will need to use, both for original data
●● data transformation and/or calcula- that will be entered and for any intermediate calculations that might be
tion sections; required.
●● a results section, which can include
3. Set up areas of the spreadsheet for data entry, calculation of intermedi-
graphics;
●● text in the form of headings and
ate values (statistical values such as sums of squares, etc.), calculation of
annotations; final statistics and, if necessary, a summary area.
●● a summary section. 4. Establish the format of the numeric data if this is different from the
default values. This can be done globally (affecting the entire spreadsheet)
or locally (affecting only a specified part of the spreadsheet).
5. Establish the column widths required for the various activities.
6. Add text (labels) to identify input, intermediate formulae and output
cells. This is valuable in error-tracking and when carrying out further
Constructing a spreadsheet – start development work. Text can be entered in designated cells, or cells can be
with a simple model and extend it grad- annotated using the New Comment feature in the Review tab.
ually, checking for correct operation as
you go. 7. Enter a test set of values to use during formula entry: use a fully worked
example to check that formulae are working correctly.
8. Enter the formulae required to make all the calculations, both interme-
diate and final. Check that results are correct using the test data.
The spreadsheet is then ready for use. Delete all of the test data values and you
have created your template. Save the template to a disk and it is then available
for repeated operations.
Graphics display
Most spreadsheets now offer a wide range of graph and chart options which
are easy to use, and this represents a way to examine your data sets rapidly and
comprehensively. The quality of the final graphics output (to a printer) variable
but is usually may be, sufficient for data exploration and analysis. Many of the
Preparing graphs using Excel 2010 –
default options are business graphics styles but there are usually histogram, bar,
Box 48.2 on p. 448 gives step-by-step
column and pie charts, X–Y plotting, line and area graphics options available.
guidance for plotted curves, histograms
Note that some spreadsheet graphics often do not come up to the standards
and pie charts.
expected for the formal presentation of scientific data, unless you manipulate
the option settings appropriately.
Printing spreadsheets
This is usually a straightforward, menu-controlled procedure, made difficult
only by the fact that your spreadsheet may be too big to fit on one piece of
paper. Try to develop an area of the sheet which contains only the data that
you need to print, i.e. perhaps a summary area. Remember that columns and/or
rows can usually be hidden for printing purposes and you can control whether
the printout is in portrait or landscape mode, and for page size and format. Use
a screen preview option, if available, to check your layout before printing. A
‘print to fit’ option is also available in some programs, making the output fit
the page dimensions.

Using spreadsheets

Frye, C. (2010) Microsoft Office Home and Student 2010 Harvey, G. (2013) Excel 2013 for Dummies. John Wiley &
Step-by-Step. Microsoft Press, Redmond. Sons Ltd, New York.
Hart-Davies, G. (2010) Beginning Microsoft Office 2010.
Apress, New York.
Study exercises
The instructions and tips for these problems assume 45.2 Create a spreadsheet and graph (advanced).
that you have Excel (2003 or later) available. If not, they
(i) Copy the data in the table below into a spread-
should be readily modified for most advanced spread-
sheet. Name and save the file appropriately.
sheet programs. If you have problems with any of the
tasks, consult Box 49.2 or try using the program’s Help Decomposition of N2O5
facility.
45.1 Create a spreadsheet and graph (introductory). Time (s) [n2O5]

0 1.000
(i) Copy the information in the table below into
a spreadsheet. Name and save the spread- 40 0.762
sheet file appropriately. 80 0.580
(ii) From the copied information, create a pie
120 0.442
chart using the Chart Wizard function.
(iii) Adjust the colours selected so the chart will 160 0.336
print out in black and white. Save the final 200 0.256
version of your spreadsheet. Print the chart
400 0.066
out directly from Excel.
600 0.017
Relative percentage composition of elements in
the Earth’s crust. (ii) Use the spreadsheet and chart-making facili-
ties to explore by eye which of the following
Element Percentage
transformations would result in the best lin-
oxygen 46.2 ear fit for these data: reciprocal, square root,
silicon 28.2 cube root or log.
(iii) Add a linear trend-line to the chart for the
aluminium 8.2
most appropriate transformation.
iron 5.6 (iv) Copy the graph to a file in Word and print out.
calcium 4.2
45.3 Use a spreadsheet as a simple database. Copy
sodium 2.4
the data in the table on p. 431 into cells within a
magnesium 2.3 spreadsheet. Modify the column widths so you
potassium 2.1 can see all of the text on a single screen. Now
sort the data in the following ways:
titanium 0.6
hydrogen 0.2 (a) by subject, in alphabetical order;
(b) by date and then by time of day;
Total 100
(c) by topic, in reverse alphabetical order.

Using spreadsheets
Use the Column Hide function (Format menu in Microsoft Excel) so that the information in columns 4 and
6 is not displayed. Find out how to undo this operation.
My exam timetable
Subject Date Time Paper Location Question style

Physical Chemistry 3 Jun Morning 1 Great Hall Multiple choice
Physical Chemistry 17 Jun Morning 2 Exam Hall 5 Written paper
Organic Chemistry 3 Jun Afternoon 1 Small Hall Short answer questions
Organic Chemistry 14 Jun Afternoon 2 Exam Hall 5 Written paper
Inorganic Chemistry 4 Jun Morning 1 Small Hall Short answer questions
Inorganic Chemistry 1 Jul Afternoon 2 Exam Hall 3 Written paper
Analytical Chemistry 13 Jun Afternoon 1 Exam Hall 5 Written paper
Analytical Chemistry 2 Jun Morning 2 Main Laboratory Practical exam

46 Using word processors, databases
and other packages
You will probably be familiar with a range of computer programs used for
Reassess your IT skills and knowledge
academic work, and may own a laptop or PC with this software installed. To
in relation to your course – although
support your studies, you will also have access to networked PCs and associ-
you may be familiar with ‘office’ style
ated programs licensed for your use. However, the specific software and ver-
software (e.g. Microsoft Office Apple
sions may vary between these systems, involving different menu protocols and
Mac Pages or Apache OpenOffice), you
commands for the same or similar functions. In addition, the specific demands
may be required to use features in new
of academic work may involve the use of sophisticated features or even new
or different ways. For example:
types of program. This chapter considers some common software applications
●● You may be asked to present docu- in relation to relevant tasks. Spreadsheets and presentation software are con-
ments and their components in very sidered in detail in Chapters 45 and 67.
specific formats (such as defining
fonts, line spacing, paragraph lay-
out, margins, layout of tables and
figures). Key Point Even when you are familiar with the basics of
●● You may benefit from using certain a software program, it may be valuable to learn more about
editing features to refine your writ- advanced features, to ensure that you are working efficiently
ing (such as word counts, thesaurus, and effectively.
spelling and grammar checker).
●● You may need to adapt your meth-
ods to longer writing exercises (such
Word processors
as creating an outline structure, writ-
ing parts out of sequence, creating a Word processors are generally available as part of an ‘office’ package with
bibliography at the outset and con- the advantage that they share a common interface in the different components
tributing to it as you write). (word processor, spreadsheet, database, etc.), allowing exchange of informa-
tion (e.g. text, graphics) between applications. Examples include Pages (Apple
Mac) and Microsoft Word (Microsoft Office). Compatible open source software
is also available, such as OpenOffice. Most word processors have similar gen-
eral features but differ in operational detail. They offer specific advantages for
academic work. For example, you can:
●● refine material many times before submission;
●● insert material easily, allowing writing to take place in any sequence;
●● use a spellchecker to check your text;
●● use a thesaurus when composing your text;
●● carry out ongoing checks of the word count;
●● produce high-quality final copies;
●● reuse part or all of the text in other documents.
Although use of computers to compose text is now almost universal, it is
worth remembering certain disadvantages of this approach:
●● the need for a reliable battery or mains power supply (important for work
in the field);
●● the need to learn specific operational details of the program;
●● the temptation to make trivial revisions, hence ‘overworking’ your text;
●● the risk of losing files or forgetting to save work appropriately.

Using word processors, databases and other packages
Key Point It is vital to save your work frequently to a memory

stick, hard drive or network drive. This should be done every
10 min or so. If you do not save regularly, you may lose hours
or days of work. Most programs can be set to ‘autosave’ every
few minutes – make sure you switch this feature on.
When using a word processor, take full advantage of the differences between
Using online help, tutorials and text-
word processing and handwriting (which necessarily follows a linear sequence
books – support information is usually
and requires more planning):
provided in one or more of the follow-
ing ways: as a help facility within the ●● Simply jot down your initial ideas for a plan, preferably at paragraph topic
program; as a file on the installation level. The order can be altered easily and if a paragraph grows too much it
CD; or as an online help support site. can easily be split.
If unfamiliar with the software, it may
●● Start writing wherever you wish and fill in the rest later.
be worthwhile investing in one of the
commercial textbooks written to sup- ●● Just put down your ideas as you think, confident in the knowledge that it is
port users of specific programs. the concepts that are important to note; their order and the way you express
them can be adjusted later.
●● Do not worry about spelling and use of synonyms – these can (and should)
be checked during a separate scan of your text, using the spell-checker first
to correct obvious mistakes, then the thesaurus to change words for style or
meaning.
●● Use a draft printout to check (a) for pace and spacing (b) to ensure that
words checked for spelling fit the required sense.
Laying out (formatting) your document

The appearance of a document on-screen generally represents what the printout
Controlling widow and orphan lines –
on paper will look like, but this should be checked using the ‘print preview’ fea-
single lines or pairs of lines at the top
ture before printing large documents, or where the paper size is non-standard.
or bottom of a page are referred to as
Menu commands can be used to adjust page margins, headers and footers, line
‘widows’ or ‘orphans’ respectively and
the TAB or M key rather than the space bar, but you can also indent or bullet
spacing and fonts, as required. Paragraph indentations should be created using
they are generally undesirable in page
design. You can control whether they
whole blocks of text using special menu options. Although you can reformat
appear in most word processors using
your text at any time, it is good practice to enter the relevant details in the Page
the appropriate commands for your
Setup when you start writing: entering them later can lead to problems due to
program.
forced reorganisation of the text layout. If you use a particular set of layout
formats regularly, e.g. an A4 page with space for a letterhead, create a formatted
template that can be called up whenever you start a new document. Advanced
users can set up automated styles and formatting features to suit particular
requirements (e.g. using the Home tab, Styles group in Microsoft Word).
Editing features
Using a spell check facility – do not
rely on this to spot all errors. Remem-
Word processors usually have many features designed to make editing doc-
ber that spell-check programs do not
uments easy. Most programs allow you to customise the toolbar to add com-
correct grammatical errors, not do they
monly used command ‘buttons’ – some examples relevant to scientific writing
pick up words that are incorrect but
include subscripts and superscripts, word count, A-Z sort and table column
that are legitimate words, e.g. ‘their’ for
formatting. The spell-checkers and thesaurus are self-explanatory, but they
‘there’, or ‘form’ for ‘from’.
generally do not include technical terms and, unless set appropriately, usually
suggest US spellings as default. Therefore, where a misspelling is‘incorrectly’

indicated (in Microsoft Word, by a red wavy underline) you may wish to add
Language setting – remember to set
the word to the dictionary file.
up your document language before you
Grammar checkers are designed to highlight text that may be incorrectly
start compiling text: this will ensure that
structured (in Microsoft Word, this is shown by a green wavy underline).
the spell-check and thesaurus provide
In general, these features are unreliable for scientific writing except when
the correct spellings by default within
pointing out obvious errors, such as a missing verb. For example, academic
your documents.
style frequently demands the use of impersonal language involving passive
tense and this is usually marked up as a matter for you to review, when it is
perfectly acceptable. Note that the grammar-check tool is customisable in
most programs.
The ability to move blocks of text (‘cut and paste’) is very valuable when
reviewing and editing, and indeed can be said to have changed the nature of
writing: it is now easy to write sections out of sequence, as availability of
information or personal interest determines. However, there is an associated
risk that the planning phase of writing (Chapter 68) is neglected, resulting in
Quick commands for commonly used weak structure. To counteract this, you should create a plan and use temporary
functions – most word processors pro- or permanent headings to organise your writing.
vide several routes to achieve the same An extremely useful editing facility is the ‘find’ or ‘search’ procedure,
end result. These include: standard which allows you to scan quickly through a document looking for a specified
menus and commands; customisable word, phrase or punctuation (in Microsoft Word, available from the Home tab,
toolbar commands; and key combina- Editing group). This is particularly valuable when combined with a replace
tions. Some useful examples of key facility so that, for example, you could replace the word ‘test’ with ‘trial’
combinations in Microsoft Word and throughout your document simply and rapidly. Format Painter is a convenient
other programs include: Ctrl + F for function that allows you to copy the format of one section to another, without
find; Ctrl + X for delete, Ctrl + P for altering the text (in Microsoft Word, available on the Home tab, Clipboard
print, Ctrl + C for copy and Ctrl + V menu). Most users will be familiar with the ‘undo’ command to rectify mis-
for paste. takes, but the ‘redo’ or ‘repeat’ command is equally valuable where repeated
formatting or text entry is required.
Fonts and line spacing

The instructions for assignments will often define specific fonts and line spac-
ing. The terminology involved is explained below.
●● Typeface: the term for a family of characters of a particular design,

each of which is given a particular name. The most commonly used for
Presenting your documents – it is good normal text is Times Roman (as used here for the main text) but many
practice not to mix typefaces too much others are widely available, particularly for the better quality printers
in a formal document; also the font size They fall into three broad groups: serif fonts with curves and flourishes
should not differ greatly for different at the ends of the characters (e.g. Times Roman); sans serif fonts without
headings, subheadings and the text. such flourishes, providing a clean, modern appearance (e.g. Arial); and
decorative fonts used for special purposes only, such as the production
of newsletters and notices.
●● Size: measured in units called points. There are about 28 points per cm.
The standard sizes for text are 10, 11 and 12 points, but typefaces are often
available up to 72 point or more.
●● Appearance: many typefaces are available in a variety of styles and weights
Many of these are not designed for use in scientific writing, but for desktop
publishing.
●● Spacing: double or 1.5 line spacing may be specified to allow the marker
to add comments or corrections. Character spacing is usually defined by the
font type (‘proportional’ spacing, where each character takes up a different

amount of space, being more readable). There are conventions for spacing
Preparing final documents – if no spe-
after each sentence – a double space used to be the norm but single is now
cific guidance is provided, use a 12-point
common. Paragraph indentation may be specified – a common format is not
proportional serif font, such as Times
to indent the first paragraph in a section, but to indent thereafter (as here),
Roman. At this font size, 1.5 line spac-
but increasingly ‘blocked’ paragraphs are used with no indents.
ing will generally be sufficient, unless
otherwise specified. You can add con- ●● Justification: this is the term describing the way in which text is aligned
trast to headings by using a sans-serif vertically. Left justification is normal, but for formal documents, both left
typeface, such as Arial, at 11 point bold. and right justification may be used (as here).
You may wish to add extra blank lines
Commands for the above features may be accessed from the Home menu
before headings and additional half
in Microsoft Word.
lines between paragraphs to improve
readability by spacing out the text.
Table construction
Tables are widely used in the biosciences, but the conventions for their pres-
entation are very specific (see Chapter 50). So, while you can produce basic
tables via tabbing or by accessing the standard formats available on most word
processor menus, this is rarely appropriate for formal submissions. Your main
options here are to:
●● Use the in-built word processor table-constructing commands. Here, the
basic table outline is constructed for you (e.g. four columns by ten rows) and
can be adjusted thereafter to fit your presentational needs (e.g. by changing
column widths, merging cells or deleting boundary lines).
●● Use a spreadsheet to construct the table and then copy it to the word
processor file. This requires considerably more manipulation than using
the word processor directly and is best reserved for special circumstances,
such as very large or complex tables of data, and especially where calcula-
tions or recalculations are involved and/or the data are already stored as a
spreadsheet.
Box 50.2 provides further guidance about how to create such tables to suit
your requirements using Microsoft Office programs.
Special characters, equations and graphics

You can draw lines and insert other small-scale graphical features such as
Inserting special characters – a wide
brackets directly within most word processors, and special characters (e.g.
range of symbols, including Greek
mathematical symbols and Greek characters) should also be available. Word
letters, and characters from other lan-
processors can be used to create simple charts and diagrams, but it is better
guages is available within most word
to create graphs using a more flexible and fully featured application (e.g. a
processing programs.
spreadsheet, see Box 50.2) then import the final output into the text of a doc-
ument. Files must be compatible, as with ‘office’ suites, but if this is so, it
is a relatively straightforward procedure. Microsoft Equation Editor may be
available on your setup to assist with mathematical formulae – consult the help
menu for guidance.
Printing
If more than one printer is attached to your PC or network, you will need to
specify which one to use from the word processor’s print menu. Most printers
offer choices as to text and graphics quality, so choose draft (low) quality for
all but your final copy since this will save both time and materials. Use a print
preview option to show the page layout if it is available. Assuming that you
have entered appropriate layout and font commands, printing should then be a

straightforward operation. Although document layout settings are modifiable

Using the print preview mode – this
at any time, changing the page, margin and font sizes will cause your text to be
can reveal errors of several kinds, e.g.
rearranged, and this can be frustrating if you have spent time carefully laying
spacing between pages, that can pre-
out the pages.
vent you wasting paper and printer ink
unnecessarily.
Databases
A database is an electronic filing system whose structure is similar to a manual
record-card collection. Its collection of records is termed a file. The individual
Definition items of information on each record are termed fields. Once the database is
constructed, search criteria can be used to view files through various filters
Relational database – effectively, a set according to your requirements. The computerised catalogues in your library
of structured tables of data where infor- are just such a system; you enter the filter requirements in the form of author
mation from each array of information is or subject keywords.
formally related to others. The database You can use a database to catalogue, search, sort and relate collections of
can be used to access and construct new information. The benefits of a computerised database over a manual cardfile
data sets without the need to adjust the system are:
original tables.
●● the information content is easily amended or updated;
●● printout of relevant items can be obtained;
●● it is quick and easy to organise through sorting and searching/selection
criteria, to produce subgroups of relevant records;
●● record displays can easily be redesigned, allowing flexible methods of
presenting records according to interest;
●● relational databases can be combined, giving the whole system immense
flexibility. The older ‘flat-file’ databases store information in files that can
Choosing between a database and a be searched and sorted, but cannot be linked to other databases.
spreadsheet – use a database only
Simple database files can be constructed within spreadsheets using the
after careful consideration. Can the
columns and rows as fields and records respectively. These programs are capa-
task be done within a spreadsheet? A
ble of reasonably sophisticated sorting, searching and filtering operations and
database program can be complex to
are probably sufficient for the types of database you are likely to require as an
set up and usually needs to be updated
undergraduate. You may also make use of a bibliographic database specially
regularly.
constructed for that purpose (e.g. EndNote, see Chapter 41).
Statistical analysis packages

Statistical packages vary from small programs designed to carry out very
specific statistical tasks to large sophisticated packages (e.g. SYSTAT, Sig-
maStat, SPSS, etc.) intended to provide statistical assistance, from experi-
mental design to the analysis of results. Consider the following features when
selecting a package:
●● The data entry and editing section – this should be user-friendly, with
options for transforming data.
●● Data exploration options – these should include descriptive statistics and
techniques for exploratory data analysis.
●● Hypothesis-testing techniques – these should include ANOVA, regres-
sion analysis, multi-variate techniques, and parametric and non-parametric
statistics.

●● Support for research methods – the program should provide assistance

Using spreadsheet statistics functions –
with experimental design and sampling.
before using a specific statistics pack-
age, check whether your spreadsheet is ●● Output facilities – these should be suitable for graphical and tabular formats.
capable of carrying out the form of anal-
Some programs have very complex data entry systems, limiting the ease of
ysis you require, as this can often be the
using data in different tests. Ideally, the data entry and storage system should be
simpler option.
based on a spreadsheet system, so that subsequent editing and transformation
operations are straightforward.
Key Point Make sure that you understand the theoretical

basis for your statistical test and the computational techniques
involved before using a particular program function.
Graphics and presentation packages

Microsoft Office programs or similar open source products can be used to
Presentation using computer packages achieve most coursework tasks requiring images, presentation slides or web
– though many computer programs pages: PowerPoint is useful for creating posters (Box 66.1) and for oral pres-
enhance presentational aspects of your entations (Box 67.1). For more advanced tasks, additional software may be
work, there are occasions when they available on your network; for example:
can make your presentation worse.
Take care to avoid the following com- ●● SigmaPlot can produce graphs with floating axes;
mon pitfalls: ●● Adobe Illustrator is useful for designing complex graphics;
●● Default or ‘chart wizard’ settings for ●● Adobe Dreamweaver enables you to produce high quality web pages;
graphs may result in output that is
unacceptable for the sciences (see ●● MindGenius can be used to produce mind maps.
Box 50.2).
Important points regarding the use of such packages are:
●● Fonts in labels and legends may not
be consistent with other parts of ●● the learning time required for some of the more complex operations can
your presentation. be considerable;
●● Some basic programs cannot pro-
duce Greek symbols (e.g. m); do not ●● the quality of your printer will limit the quality of your output;
use ‘u’ as a substitute. The same
●● not all files will readily import into a word processor you may need to save
applies to scientific notation and
superscripts: do not use 14C for 14C, your work in a particular format. The different types of file are distinguished
and replace, e.g., 1.4 E + 09 with by the three-character filename extension, e.g. .jpg and .bmp.
1.4 * 109. First try cutting and past-
ing symbols from Microsoft Word
or, if this fails, leave space and draw Key Point ‘Default’ computer graphics are rarely satisfactory
the correct symbols by hand on the for scientific presentation. Make appropriate changes to suit sci-
printout. entific standards and style. Box 70.1 gives a checklist for graph
drawing and Box 70.2 provides guidelines for adapting Micro-
soft Excel output.
Image storage and manipulation

With the widespread use of digital images, programs that facilitate the storage
and manipulation of electronic image files have become increasingly important.
These programs create a library of your stored images and provide a variety
of methods for organising and selecting images. Programs for image manip-
ulation vary widely in capability, cost and associated learning time. Adobe
Photoshop and Paint Shop Pro are two widely available examples. Many are

highly sophisticated programs intended for graphic artists. For most scientific
Take care when copying images – you
purposes, however, only relatively limited functions are required, such as image
may be at risk of committing plagiarism
cropping and resizing.
and copyright infringement (Chapter 42).

Apache. OpenOffice.org. Available: http://www.openoffice Gookin, D. (2013) Word 2013 for Dummies. John Wiley &
.org/ Sons Ltd, New York.
Last accessed 01/09/15. Wang, W. (2013) Office 2013 for Dummies. John Wiley &
[Website containing information and links for down- Sons Ltd, New York.
loading freeware ‘office’ type programs.]
[And similar texts for other packages and release
Fuller, L.U., Cook, K. and Kaufeld, J. (2013) Access 2013 versions.]
for Dummies. John Wiley & Sons Ltd, New York.
Study exercises
46.1 Investigate intermediate/advanced Word fea- meaning. Try, for example, to find alterna-
tures. The tasks in the following list are likely to tives to the word ‘alternative’.
be useful in preparing assignments and report (h) Carry out a spell-check on your document.
writing within chemistry. Can you carry out all (i) Carry out a word count on your document
of the tasks? If not, use either a manual or thy and on a selected part of it.
online Help feature to find out how to accomplish (j) Open two documents and switch between
them. Tips are given in the answer section. them.
(a) Sort information in a list into alphabetical 46.2 Make precise copies of tables. Copy the follow-
order. ing tables using word processor software.
(b) Replace a text string word or phrase with a Test organism Results of analysis (units)
new text string throughout your document.
August September October
(c) Replace a text string in normal font with the
same text string in italics throughout your X
document. Y
(d) Add a ‘header’ and ‘footer’ to your docu-
Z
ment, the former showing the document’s
title and the latter containing page numbers
Results of analysis (units)
on the bottom centre of the page.
(e) Adjust the margins of the page to give a 5-cm Test organism August September October
margin on the left and a 2-cm margin on the X
right.
Y
(f) Change the type of bullets used in a list from
standard (• or ■) to a different form (e.g. -, Z
◆ or ✓).
46.3 Investigate what programs and packages are
(g) Use the ‘thesaurus’ option to find a differ-
available to you as a student. Test each program
ent or more suitable word to express your
with appropriate data, images, etc.

Analysis and presentation of data
47. Fundamental principles of quantitative
chemical analysis 437
48. Calibration and quantitative analysis 441
49. Using graphs 447
50. Presenting data in tables 459
51. Hints for solving numerical problems 463
52. Manipulating and transforming raw data 472
53. Descriptive statistics 476
54. Choosing and using statistical tests 487
55. Drawing chemical structures 501
56. Chemometrics 507
57. Computational chemistry 514

47 Fundamental principles of quantitative
chemical analysis
Most analytical methods rely on one or more chemical or physical properties

of the test substance (the analyte) for detection and/or measurement. There are
two principal approaches:
1. Qualitative analysis – where a sample is analysed to determine whether
Definitions a compound is present or absent. For example, the use of infrared spec-
troscopy (Chapter 36) and nuclear magnetic resonance spectroscopy
Accuracy The closeness of an indi-
(Chapter 37).
vidual measurement, or a mean value
based on a number of measurements, 2. Quantitative analysis – where the quantity of a particular element or com-
to the true value. pound in a sample is determined in terms of its concentration (e.g. as
Concentration range The range of val- mg L-1). For example, the use of atomic spectroscopy (Chapter 30) and
ues from the detection limit to the upper X-ray fluorescence spectroscopy (Chapter 31) for elemental analysis, or
concentration at which the technique gas and liquid chromatography (Chapter 32) for compound analysis.
becomes inaccurate or imprecise. Your choice of approach will be determined by the purpose of the investi-
Detection limit The minimum concen- gation and by the level of accuracy and precision required. Many of the basic
tration of an analyte that can be detected quantitative methods rely on chemical reactions of the analyte and involve
at a particular confidence level. assumptions about the nature of the test substance and the lack of interfering
Drift ‘Baseline’ movement in a par- compounds in the sample: such assumptions are unlikely to be wholly valid
ticular direction: drift can be a problem at all times. If you need to make more exacting measurements of a particular
during analysis (e.g. when separating analyte, it may be necessary to separate it from the other components in the
compounds by chromatography). sample, e.g. extraction of the analyte using solid phase extraction to remove
interferences (Chapter 32), using chromatography (Chapter 32) or electropho-
Noise Random fluctuations in a contin-
resis (Chapter 33), and then identify the separated components, e.g. using mass
uously monitored signal.
spectrometry (e.g. Chapter 38).
Precision The extent of mutual agree-
ment between replicate data values for
an individual sample.
Key Point In general, you should aim to use the simplest pro-
Quality control Measures in place to cedure that satisfies the purpose of your investigation – there
ensure that the result meets your labo- is little value in using a complex, time-consuming or costly
ratory’s standard. analytical procedure to answer a simple problem when a high
Quality assurance Measures in place degree of accuracy is not required.
to monitor and document the perfor-
mance of a test procedure, e.g. profi-
ciency testing schemes. Most of the routine methods based on chemical analysis are destructive
Replicate Repeat measurement. since the analyte usually has to be extracted from the matrix in which it is held,
Selectivity The extent to which a for example, extracting drugs from blood or urine (Chapter 32) or decomposi-
method is free from interference due to tion of a solid sample using concentrated acids (Chapter 30). However, many
other substances in the sample. analytical methods that are based on physical properties are non-destructive,
for example compound identification using Fourier transform infrared (FTIR)
Sensitivity The ability to discriminate
spectroscopy (Chapter 36). Non-destructive methods are often preferred, as
between small differences in analyte
they allow further characterisation of a particular sample.
concentration.
Validation The process whereby the
Validity and quantitative analysis
accuracy and precision of a particular
analytical method are checked in rela- Before using a particular procedure, you should consider its possible limitations
tion to specific standards, using an in terms of:
appropriate reference material contain-
●● measurement errors, and their likely magnitude: these might include process-
ing a known amount of analyte.
ing errors (e.g. in preparing solutions and making dilutions), instrumental

Fundamental principles of quantitative chemical analysis
errors (e.g. a gas chromatography instrument that has not been set up cor-
Selected suppliers of certified reference
rectly), calibration errors (e.g. converting a digital readout to an analyte
materials – UK: LGC – Laboratory of
concentration) and errors due to the presence of interfering substances (e.g.
the Government Chemist, London.
fatty acids in blood samples);
USA: NIST – National Institute of Sci-
ence and Technology, Washington DC. ●● sampling errors: these may occur if the material used for analysis is not
representative, e.g. elemental analysis of a soil sample or drug composition
of a biological fluid (see Prichard and Barwick, 2007).
Certified reference materials can be
used for a variety of purposes including The reliability of a particular method can be assessed by measuring ‘standards’
(sometimes termed ‘controls’). These are often prepared in the laboratory by
●● establishing metrological traceabil-
adding a known amount of analyte, or reference material, to a sample matrix.
ity of results;
●● confirmation of the identity of a For example, when analysing a blood sample, controls should be prepared
material; using real blood, such as horse blood or human transfusion blood (this is often
●● development and validation of new termed ‘spiking’ a sample). The reference material used for spiking must be
methods of measurement; pure and supplied from a recognised source. These are available from com-
●● calibration and verification of mercial suppliers who will also provide a certificate of analysis indicating the
measurement processes in routine exact concentration and purity of the compound. In most instances, several
analyses; standards (including a ‘blank’ or zero) are analysed to construct a calibration
●● verification of the correct application curve (see Chapter 48), which is then used to convert sample measurements to
of standard methods; amounts of analyte.
●● internal quality control schemes;
The use of certified reference materials (CRMs) is particularly important
●● defining values for other materials
when establishing (validating) new methods or comparing methods. CRMs
which may then be used as second-
ary standards or calibrants. are obtained from recognised suppliers and can either be analysed directly or
extracted/decomposed and then analysed. As well as the actual material all
CRMs have a certificate which provides details of element/compound con-
Accreditation – In the UK, analytical
tent, in appropriate units, and an estimate of uncertainty of the content, e.g.
science laboratories can be accredited
Pb-content of 12.5 { 0.3 mg/kg. The results obtained within the laboratory
by UKAS (United Kingdom Accredita-
can then be compared with the certificate values. Ideally agreement between
tion Services) to ensure that all ana-
measured and certified values for each analyte should be obtained before con-
lytical procedures are performed to an
tinuing with the selected experimental approach.
internationally recognised standard (BS
CRMs can be obtained for a whole variety of analytes in a wide range
ISO 17025).
of matrices (chemical, biological and foodstuffs). Care should be taken in
matching both concentrations of analytes and matrix between samples and
the chosen CRM.
Validation of a particular method is particularly important in the pharma-
A proficiency testing scheme should ceutical, environmental or food analysis laboratory, where the results of the
test the: analysis have important implications. Such laboratories operate strict valida-
●● organisation tion procedures which include: (i) adherence to standard operating procedures
●● quality system (SOPs); (ii) calibration of assays using CRMs containing known amounts of
●● quality audits and reviews analyte and traceable to a national reference laboratory; (iii) effective quality
●● staff assurance and quality control systems; (iv) detailed record-keeping, covering
●● equipment all aspects of the analysis and recording of results. Such rigour is often required
●● measurement traceability for legal reasons and consumer protection, although the general principles of
●● methods and procedures
standardisation, calibration, assessment of performance and record-keeping
●● environment
are equally valid.
●● handling of samples
●● records
●● complaints
●● sub-contracting and purchasing Proficiency testing schemes
These are used by laboratories carrying out analytical measurements and
based on NAMAS criteria available at
will give a snapshot of a laboratory’s performance and quality systems at any
http://www.smtl.co.uk/MDRC/NAMAS/
one point of time. By reviewing the performance over a period of time, the
about-namas.html
analytical quality of the laboratory can be determined.
438 Analysis and presentation of data

Reporting analytical results

Criteria for the selection of a particular
analytical method – Chemical analysis demands both accuracy and precision (p. 38). In order to
●● the required level of accuracy and
report the presence of, for example, a drug in a blood sample, you have to be
precision certain that the drug is present. In order to do so, a combination of analytical
●● the number of samples to be procedures should be used. GC–MS (Chapter 32) used for screening the blood
analysed sample may identify the specific drug using a combination of the retention time
●● the amount of each sample available and the mass spectrum of the drug. Using this procedure, an internal standard
for analysis (p. 271) is added to the blood sample prior to extraction. This ensures that (i) the
●● the physical form of the samples extraction process is working satisfactorily and (ii) the instrument is operating
●● the expected concentration range of correctly. If a drug has been identified using this process, a reference standard is
the analyte in the samples analysed which should elute at the same retention time ( {2%) (Anon., 2002) of
●● the sensitivity and detection limit of the drug in the case sample. Occasionally, after the extraction process, the extract
the technique
will have to be derivatised prior to analysis by GC–MS, for example, mor-
●● the lik elihood of interfering
substances
phine is usually derivatised using N-methyl-N-trimethylsilyltrifluoroacetamide
●● the speed of the analysis (MSTFA). If a drug has been derivatised, it can still be identified by the com-
●● the ease and convenience of the bination of retention time and mass spectrum in comparison to a derivatised
procedure reference standard. Once the drug has been identified, the presence of the drug
●● the skill required by the operator can either be confirmed or quantified, usually by a technique based on a different
●● the cost and availability of the chemical principle to that used for screening, such as HPLC (Chapter 32) or
equipment. LC–MS (Chapter 38). It is only after the process of identification, quantification
(if appropriate) and confirmation that the drug can be reported.
Text references
Anon. (2002) Forensic Toxicology Laboratory Guidelines. Prichard, E. and Barwick, V. (2007) Quality Assurance in
Society of Forensic Toxicologists, SOFT. Analytical Chemistry. John Wiley & Sons Ltd, Chichester.

Anon. (2004) Guidance on Analytical Methods Validation. Currell, G. (2000) Analytical Instrumentation. Performance
Available: http://www.fda.gov/cber/gdlns/methval.htm and Characteristics of Quality. John Wiley & Sons Ltd,
Last accessed 15/12/04. Chichester.
[US FDA guidance/details of reference standards, proce- Funk, W., Dammann, V. and Donnevert, G. (2007) Quality
dures for method validation, etc.] Assurance in Analytical Chemistry. 2nd edn. John Wiley &
Anon. (2004) Valid Analytical Measurement Homepage. Sons Ltd, Chichester.
Available: http://www.vam.org.uk Kellner, R., Mermet, J.M., Otto, M., Valcarcel, M. and
Last accessed: 15/12/04. Widner, H.M. (2004) Analytical Chemistry: A Modern
Barwick, V., Burke, S., Lawn, R., Roper, P. and Walker, R. Approach to Analytical Science, 2nd edn. John Wiley &
(2001) Applications of Reference Materials in Analytical Sons Ltd, Chichester.
Chemistry. RSC, Cambridge. Konieczka, P. and Namiesnik, J. (2009) Quality Assur-
Burgess, C. (2000) Valid Analytical Methods and Proce- ance and Quality Control in the Analytical Chemical
dures. Royal Society of Chemistry, Cambridge. Laboratory. A Practical Approach. CRC Press, Boca
Chan, C.C., Lee, Y.C., Lam, H. and Zhang, X-M. (eds) Raton.
(2004) Analytical Method Validation and Instrument Prichard, E. and Barwick, V. (2007) Quality Assurance
Performance Verifications. John Wiley & Sons Ltd, in Analytical Chemistry. John Wiley & Sons Ltd,
Chichester. Chichester.

Ratliff, T.A. (2003) The Laboratory Quality Assurance Sys- Stoeppler, M., Wolf, W.R. and Jenks, P.J. (eds) (2001) Ref-
tem. A manual of quality procedures and forms, 3rd edn. erence Materials for Chemical Analysis. John Wiley &
John Wiley & Sons Ltd, Chichester. Sons Ltd, Chichester.
Schwedt, G. (1998) The Essential Guide to Analytical Swartz, M.E. and Krull, I.S. (2012). Handbook of Analytical
Chemistry. John Wiley & Sons Ltd, Chichester. Validation. CRC Press, Boca Raton.
Study exercises
47.1 Test your knowledge of quality in analytical 47.3 Based on your knowledge of an instrumental
measurements – explain the meaning of the analytical technique that you have used consider
following terms and why they are important in the following issues:
quantitative chemical analysis:
(a) What physical form must the sample be in for
(a) accuracy the technique to accept the sample?
(b) precision (b) For quantitative analysis what is the expected
(c) quality control concentration range for the calibration?
(d) quality assurance (c) Does the technique suffer from any known
(e) replicate interferences?
(f) detection limit. (d) How would you dilute or pre-concentrate the
analyte in the sample?
47.2 Offer your opinion – discuss with your colleagues
whether or not all analytical chemistry laborato-
ries should be accredited to UKAS standards.

48 Calibration and quantitative analysis
There are many instances where it is necessary to measure the quantity of a test
substance using a calibrated procedure. You are most likely to encounter this
approach in one or more of the following practical exercises:
●● Quantitative spectrophotometric assay (Chapter 29).
●● Atomic absorption spectroscopic analysis of metal ions in quantitative
analysis of metal ions in solution using atomic spectroscopy (Chapter 30)
Understanding quantitative measure- or electroanalytical techniques (Chapter 34).
ment – Chapter 47 contains details of
the basic principles of valid measure-
●● Quantitative chromatographic analysis (e.g. Chapter 32).
ment, while Chapters 29–40 deal with ●● Quantitative element analysis of solids using XRF (Chapter 31).
some of the specific analytical tech-
niques used in chemistry. The use of
internal standards is covered on p. 271. Key Point In most instances, calibration involves the estab-
lishment of a relationship between the measured response
(the ‘signal’) and one or more ‘standards’ containing a known
Calibrating laboratory apparatus – this
amount of substance.
is important in relation to validation of
equipment, e.g. when determining the
accuracy and precision of a pipettor by In some instances, you can measure a signal due to an inherent property of the
the weighing method: see p. 66. substance (e.g. the absorption of UV light p. 217), whereas in other cases you
will need to react it with another substance to see the result (e.g. an acid–base
titration, visualised using an indicator p. 199).
The different types of calibration curve

By preparing a set of solutions (termed ‘standards’), each containing either (i) a
(a) (b)
known amount or (ii) a specific concentration of the substance, and then meas-
uring the response of each standard solution, the underlying relationship can be
established in graphical form as a ‘calibration curve’, or ‘standard curve’. This
can then be used to determine either (i) the amount or (ii) the concentration
log y
of the substance in one or more test samples. Alternatively, the response can
be expressed solely in mathematical terms: an example of this approach is the
determination of chlorophyll pigments in plant extracts by measuring absorp-
x x tion at particular wavelengths, and then applying a formula based on previous
(published) measurements for purified pigments.
(c) (d) There are various types of standard curve: in the simplest cases, the rela-
tionship between signal and substance will be linear, or nearly so, and the
calibration will be represented best by a straight-line graph (see Box 48.1). In
some instances (Fig. 48.1(a)), you will need to transform either the x values
or y values, in order to produce a linear graph. In other instances, the straight-
y
y
line relationship may only hold up to a certain value (the ‘linear dynamic
range’) and beyond this point the graph may curve (e.g. in quantitative spec-
trophotometry the Beer–Lambert relationship often becomes invalid at high
x x
absorbance, giving a curve, Fig. 48.1(b)). Some calibration curves are sigmoid
Fig. 48.1 Calibration curves: (a) log-linear; (Fig. 48.1(c)). Finally, the signal may decrease in response to an increase in
(b) curvilinear; (c) sigmoid, or S-shaped; the substance (Fig. 48.1(d), e.g. radioimmunoassay), where an inverse sigmoid
(d) inverse. calibration curve is obtained. In some practical classes, you may be told that the

Calibration and quantitative analysis
Box 48.1 The stages involved in preparing and using a calibration curve
1. Decide on an appropriate test method – for exam- 1

ple, in a project, you may need to research the best 0.8
approach to the analysis of a particular compound in
absorbance
0.6
your sample.
0.4
2. Select either (a) amount or (b) concentration, and
an appropriate range and number of standards – in 0.2
practical classes, this may be given in your sched- 0
ule, along with detailed instructions on how to make 0 10 20 30 40 50
up the standard solutions. In other cases, you may amount (ng)
be expected to work this out from first principles
Fig. 48.2 Typical calibration curve for spectrophotometric
(Chapters 9, 30 and 32 give worked examples) – aim
analysis.
to have evenly spaced values along the x axis.
3. Prepare your standards very carefully – due attention use type linear regression (p. 499) to produce a linear
to detail is required: for example, you should ensure trend line (also termed the ’line of best fit’) and you
that you check the calibration of pipettors beforehand, can then quote the value of r2, which is a measure of
using the weighing method (p. 66). Don’t forget the the ‘fit’ of the measurements to the line (see p. 501). In
’zero standard’ plus any other controls required, for chemical analysis the value of r2 should normally be
example to test for interference due to other chemical as close to 1.00 as possible. However, you should take
substances. Your standards should cover the range of care not to use a linear plot when the underlying rela-
values expected in your test samples. tionship is clearly non-linear (p. 500) and you must
consider whether the assumptions of the regression
4. Analyse the standards and the unknown (test) analysis are valid (e.g. for transformed data).
samples – preferably all at the same time, to avoid
introducing error due to changes in the sensitivity or 6. Determine the amount or concentration in each
drift in the zero setting of the instrument with time. unknown sample – either by reading the appropri-
It is a good idea to measure all of your standard ate value from the calibration curve, or by using the
solutions at the outset, and then measure your test underlying mathematical relationship, i.e. y = a + bx
solutions, checking that the ’zero standard’ and ’top (p. 472). Make sure you draw any horizontal and ver-
standard’ give the same values after, say, every six tical construction lines very carefully – students often
test measurements. If the remeasured standards do lose marks unnecessarily by submitting poorly drawn
not fall within a reasonable margin of the previous construction lines within practical reports.
value, then you will have to go back and recalibrate 7. Correct for dilution or concentration, where appro-
the instrument, and repeat the last six test measure- priate – for example, if you diluted each test sample
ments. If your test samples lie outside the range of by ten-fold, then you would need to multiply by 10 to
your standards, you may need to repeat the assay determine the value for the undiluted test sample. As
using diluted test samples (extrapolation of your another example, if you analysed 0.2 mL of test sam-
curve may not be valid, see p. 458). ple, you would need to multiply the value obtained
from the calibration curve by 5, to give the value per
5. Draw the standard curve, or determine the underly-
mL of the original sample.
ing relationship – Figure 48.2 gives an example of a
typical linear calibration curve, where the spectropho- 8. Quote your test results to an appropriate number of
tometric absorbance of a series of standard solutions significant figures – this should reflect the accuracy
is related to the amount of substance. When using of the method used (see p. 38), not the size of your
Excel® or graphics packages, it is often appropriate to calculator’s display.
relationship is expected to be linear, curvilinear, or whatever, while in others

you may be expected to decide the form of the standard curve as part of the
exercise.
Key Point You should always aim for a linear regression value
(r 2) 7 0.99 in your calibration curve

Practical considerations
Example A test tube containing 8
mL of water plus 2 mL of 1% w/v NaCl Amount or concentration?
(Mr = 58.44) would have a mass con-
This first step is often the most confusing for new students. It is vital that
centration of 0.2% w/v NaCl
(a five-fold dilution of the original NaCl
you understand the difference between amount of substance (e.g. mg,
solution), which can also be expressed ng), and concentration (the amount of substance per unit volume, e.g.
as 2 g L-1: in terms of molar concen- mmol L-1, mol m -3, % w/v) before you begin your practical work.
tration (p. 74), this would be equiva-
lent to 2 , 58.44 = 0.0342 mol L-1 (to
three significant figures). Expressed in
terms of the amount of NaCl in the test Key Point Essentially, you have to choose whether to work in
tube, this would be 0.02 g in mass, terms of either (i) the total amount of substance in your assay
or 0.02 , 58.44 = 3.42 * 10-4 mol vessel (e.g. test tube or cuvette) or (ii) the final concentration of
(342 mmol) in moles. the substance in your assay vessel.
The interconversion of amount and concentration is covered in more detail

on p. 43. Either way, this is usually plotted on the x-(horizontal) axis and the
measured response on the y-(vertical) axis.
Choice of standards
In your early practical classes, you may be provided with a stock solution (p. 73),
Organisations providing national/
from which you then have to prepare a specified number of standard solutions.
international standards – these
In such cases, you will need to understand how to use dilutions to achieve the
include:
required amounts or concentrations (pp. 72, 74). In later work and projects, you
may need to prepare your standards from chemical reagents in solid form, where
●● Laboratory of the Government the important considerations are purity and solubility (p. 74). For professional
Chemist (http://www.lgc.co.uk/) for analysis (e.g. in forensic science or analytical science), it is often important to
UK standards and European Refer- be able to trace the original standard or stock solution back to national or inter-
ence Materials.
national standards (Chapter 47) or to certified reference materials.
●● National Institute for Science and
Technology (http://www.cstl.nist How many standards are required?
.gov/), for biochemical/chemical
standards in the USA.
This may be given in your practical schedule, or you may have to decide what
●● Institute for Reference Materials and is appropriate (e.g. in project work and research). If the form and working range
Measurements (http://www.irmm.jrc of the standard curve is known in advance, this may influence your choice –
.be/) for European standards, includ- for example, linear calibration curves can be established with fewer standards
ing BCR and ERM materials. than curvilinear relationships. In some instances, analytical instruments can be
●● OIE Biological Standards Commis- calibrated using a single standard solution, often termed a single point ‘cali-
sion (http://www.oie.int/bsc/eng/ brator’. Replication of each standard solution is a good idea, since it will give
en_bsc.htm), for international animal you some information on the variability involved in preparing and assaying
standards. the standards. Consider whether you should plot mean values on your standard
curve, or whether it is better to plot the individual values (if one value appears
to be well off the line, you are likely to have made an error, and you may need
to check and repeat the measurement).
Preparing your standards
It is extremely important to take the greatest care to measure out all chemicals
Plotting a standard curve – do not
force your calibration line to pass
and liquids very accurately, to achieve the best possible standard curve. The grade
through zero if it clearly does not. There
of volumetric flask used and temperature of the solution also affect accuracy
is no reason to assume that the zero
(grade A apparatus is best). You may also consider what other additives might
value is any more accurate than any
be required in your standard solutions. For example, do your test samples have
other reading you have made.
high levels of potentially interfering substances, and should these also be added
to your standards? Also consider what controls and blank solutions to prepare.

Box 48.2 How to use a spreadsheet (e.g. a Microsoft Excel) to produce a linear regression plot
Two approaches are possible: either using the trendline 1

feature or using the regression analysis function within
0.8 y = 0.0155x + 0.0352
the data analysis took pack. Both make the assumption
R 2 = 0.9836
that the criteria for Model I linear regression are met (see
Absorbance
0.6
p. 499). In the example shown below, the following sim-
ple data set has been used: 0.4
0.2
Amount (ng) Absorbance
0
0 0.00 0 20 40 60
Amount (ng)
10 0.19
20 0.37 Fig. 48.3 Calibration curve showing line of best fit and
details of linear regression equation.
30 0.56
40 0.63
and use the Format Trendline 7 Line style menu
50 0.78 to adjust the Width of the line. Drag and move the
equation panel if you would like to alter its location
Using the Trendline feature. This quick method provides on the chart, or delete it, having noted the values.
a line of best fit on an Excel chart and can also provide a Figure 48.3 shows a calibration curve produced in this
set of equation values for predictive purposes. way for the data presented above.
1. Create a graph (chart) of your data. Enter the data in 4. Use the regression equation to estimate unknown
two columns within your spreadsheet, select the data (test) samples. By rearranging the equation for a
array (highlight using left mouse button) and then, straight line and substituting a particular y value, you
using the Insert tab Charts group, select Scatter. can predict the amount/concentration of substance (x
value) in a test sample. This is more precise than sim-
2. Add a trend line. Right-click on any of the data points on ply reading the values from the graph using construc-
your graph, and select the AddTrendline menu. Choose tion lines (Box 48.1). If you are carrying out multiple
the Linear trend line option, but do not click OK at this calculations, the appropriate equation, x = (y - a)/b,
stage. Rather, from the Trendline Options menu, select: can be entered into a spreadsheet, for convenience.
(i) Display equation on chart and (ii) Display R-squared
value on chart. Now click Close. The equation (shown Using the Regression Analysis tool requires the ‘data
in the form y = bx + a) gives the slope (b) and inter- analysis took pack’ to be loaded beforehand (using the
cept (a) of the line of best fit, while the R-squared value File tab, Options menu, Add-Ins, then select Analysis
(coefficient of determination, p. 500) gives the propor- ToolPak) and this provides summary output that contains
tional fit to the line (the closer this value is to 1, the details of slope, intercept and coefficient of determina-
better the fit of the data to the line). tion along with an analysis of variance (ANOVA) table
(Chapter 54 gives further details of these aspects, includ-
3. Modify the graph to improve its effectiveness. For ing an example of output in study exercise 54.4).
a graph that is to be used elsewhere (e.g. in a lab
write-up or project report), adjust the display to Note Instructions illustrated here may vary among the
remove the default background and gridlines and different versions of Microsoft Office programs. Check
change the symbol shape (see Box 54.2 for more the exact functions and syntax using the Insert Function
advice and examples). Right-click on the trend line menu option (fx button in some versions).
Key Point The validity of your standard curve depends upon

careful preparation of standards, especially in relation to accu-
rate dispensing of the volumes of any stock solution and diluting
liquid (diluent) – the results for your test samples can only be as
good as your standard curve.

Preparing the calibration curve and determining the amount

Dealing with interfering substances –
one approach is to use the method of
of the unknown (test) sample(s)
’standard additions’, where the stand-
This is described in stepwise fashion in Box 48.1. Check you understand
ards all contain a fixed additional
the requirements of graph drawing, especially in relation to plotted curves
amount of the sample (for more details
(p. 453) and the mathematics of straight-line graphs (p. 472). Spreadsheet pro-
of this approach (see e.g., Dean, 1997).
grams such as Microsoft Excel can be used to produce a regression line for a
Internal standards (p. 271) can also be
straightline calibration plot (p. 446). Examples of how to do this are provided
used to detect such problems.
in Box 48.2.
Text reference
Dean, J.D. (1997) Atomic Absorption and Plasma Spec-
troscopy, 2nd edn. John Wiley & Sons Ltd, Chichester.

Cable, M. (2005) Calibration: A Technician’s Guide ISA, Harris, D.C. (2010) Quantitative Chemical Analysis, 8th
Raleigh. edn. Freeman, New York.
Miller, J.N. and Miller, J.C. (2010) Statistics and
Chemometrics for Analytical Chemistry, 6th edn. Pren-
tice Hall, Harlow.
Study exercises
48.1 Determine unknowns from a calibration curve Give your answers to three significant figures in
produced in Excel. The following data are for a each case.
set of calibration standards for chlorophenol in
48.2 Identify the errors in a calibration curve. The fig-
a sample extract.
ure below shows a calibration curve of the type
Concentration (mg L −1) Peak area that might be submitted in a practical write-up.
List the errors and compare your observations
0 0 with the list given on p. 454.
2 14 567
0.45
4 30 124
0.4
6 45 623 0.35
8 60 021 0.3
Absorbance
10 71 209 0.25
15 112 458 0.2

0.15
Using PC-based software (e.g. Excel), fit a trend
0.1
line (linear regression) and determine the chloro-
0.05
phenol content of the following extracted water
samples with the following peak areas: 0
2000 4000 6000 8000 10000
(a) 8 741, (b) 23 478, (c) 38 500. Amount (ng)
(continued)

Study exercises (continued)
6000
20 2054
y = 101.04x + 16
5000
30 3089 R 2 = 0.9995
40 4100 4000
Peak area
50 5007 3000
2000
48.3 Using the Trendline feature. This quick method
provides a line of best fit on an Excel chart and 1000
can also provide a set of equation values for pre-
0
dictive purposes. 0 10 20 30 40 50 60
Concentration (µg/mL)
1. Create a graph (chart) of your data. Enter
the data in two columns within your spread- Fig. 48.4 Calibration curve showing line of best fit and
sheet, select the data array (highlight using details of linear regression equation.
left mouse button) and then, using the ‘Insert’
icon, select ‘Scatter’ and then the icon ‘Scat- the Format Trendline 7 line style menu to
ter only with markers’. adjust the Weight of the line to make it thin-
ner or thicker. Drag and move the equation
2. Add a trend line. Right-click on any of the
panel if you would like to alter its location on
data points on your graph, and select the
the chart. Fig. 48.4 shows a typical calibration
Add Trendline menu. Choose the Linear trend
curve produced in this way.
line option, but do not click OK at this stage.
Select: (i) Display equation on chart and 4. Add a title and axes labels. Click on the Lay-
(ii) Display R-squared value on chart. Now out icon 7 chart title (to add a title) and Lay-
click OK. The equation (shown in the form out icon 7 chart axis (horizontal or vertical to
y = bx + a) gives the slope and intercept of add a label to the x and y axis respectively).
the line of best fit, while the R-squared value
5. Use the regression equation to estimate
(coefficient of determination, p. 499) gives
unknown (test) samples. By rearranging
the proportional fit to the line (the closer this
the equation for a straight line and substitut-
value is to 1, the better the fit of the data to
ing a particular y-value, you can predict the
the trend line).
amount/concentration of substance (x-value)
3. Modify the graph to improve its effective- in a test sample. This is more precise than
ness. For a graph that is to be used else- simply reading the values from the graph
where (e.g. in a lab write-up or project report), using construction lines. If you are carrying
adjust the display to remove the default back- out multiple calculations, the appropriate
ground and gridlines and change the symbol equation, x = (y - a)/b, can be entered into
shape. Right-click on the trend line and use a spreadsheet, for convenience.

49 Using graphs
0.6 Graphs can be used to show detailed results in an abbreviated form, displaying
0.5 the maximum amount of information in the minimum space. Graphs and tables
0.4 present findings in different ways. A graph (figure) gives a visual impression
absorbance
0.3 of the content and meaning of your results, while a table provides an accurate
0.2
numerical record of data values. You must decide whether a graph should be
0.1
used, e.g. to illustrate a pronounced trend or relationship, or whether a table
(Chapter 50) is more appropriate.
0
0 2 4 6 8 10 A well-constructed graph will combine simplicity, accuracy and clarity.
concentration/mg mL-1 Planning of graphs is needed at the earliest stage in any write-up as your accom-
panying text will need to be structured so that each graph delivers the appropri-
Fig. 49.1 Calibration curve for the determi- ate message. Therefore, it is best to decide on the final form for each of your
nation of lead in soil using FAAS. Vertical
graphs before you write your text. The text, diagrams, graphs and tables in a
bars show standard errors (n = 3).
laboratory write-up or project report should be complementary, each contrib-
uting to the overall message. In a formal scientific communication it is rarely
0 100 200 300 400 500 600 700 necessary to repeat the same data in more than one place (e.g. as a table and as
0
a graph). However, graphical representation of data collected earlier in tabular
ln [N2O5] (mol L-1)
-1
-2
format may be applicable in laboratory practical reports.
-3
-4 Practical aspects of graph drawing
-5
time (s) The following comments apply to graphs drawn for laboratory reports. Figures
for publication, or similar formal presentation, are usually prepared according
Fig. 49.2 Decomposition of N2O5. The
to specific guidelines, provided by the publisher/organiser.
first-order rate constant can be determined
from the slope of the line.
Key Point Graphs should be self-contained – they should
include all material necessary to convey the appropriate mes-
Criteria for preparing graphs
sage without reference to the text. Every graph must have a con-
●● Clarity – show the relationships cise explanatory title to establish the content. If several graphs
within the data. are used, they should be numbered so they can be quoted in
●● Economy – leave out superfluous the text.
information/text.
●● Integrity – present the data without
trying to mislead. ●● Consider the layout and scale of the axes carefully. Most graphs are used
to illustrate the relationship between two variables (x and y) and have two
axes at right angles (e.g. Fig. 49.1 and 49.2). The horizontal axis is known as
Selecting a title – it is a common fault the abscissa (x axis) and the vertical axis as the ordinate (y axis).
to use titles that are grammatically incor- ●● The axis assigned to each variable must be chosen carefully. Usually the
rect. A widely applicable format is to state x axis is used for the independent variable (e.g. treatment) while the depend-
the relationship between the dependent variable (e.g. chemical response) is plotted on the y axis. When neither
ent and independent variables within variable is determined by the other, or where the variables are interdependent,
the title, e.g. ‘The relationship between the axes may be plotted either way round.
enzyme activity and external pH’.
●● Each axis must have a descriptive label showing what is represented,
together with the appropriate units of measurement, separated from the
descriptive label by a solidus or ‘slash’ (/), as in Fig. 49.1, or by brackets,
as in Fig. 49.2.
●● Each axis must have a scale with reference marks (‘tics’) on the axis to
show clearly the location of all numbers used.
●● A figure legend should be used to provide explanatory detail, including
a key to the symbols used for each data set.

Using graphs
Handling very large or very small numbers

Remembering which axis is which – a
way of remembering the orientation of To simplify presentation when your experimental data consist of either very
the x axis is that ‘x’ is ‘a cross’, and it large or very small numbers, the plotted values may be the measured numbers
runs ‘across’ the page (horizontal axis) multiplied by a power of 10: this multiplying power should be written imme-
while ‘y’ is the first letter of ‘yacht’, with diately before the descriptive label on the appropriate axis (as in Fig. 49.3).
a large vertical mast (vertical axis). However, it is often better to modify the primary unit with an appropriate prefix
(p. 40) to avoid any confusion regarding negative powers of 10.
Size
Remember that the purpose of your graph is to communicate information.
y-axis
It must not be too small, so use at least half an A4 page and design your
axes and labels to fill the available space without overcrowding any adjacent
text. If using graph paper, remember that the white space around the grid is
usually too small for effective labelling. The shape of a graph is determined
x-axis
by your choice of scale for the x- and y-axes which, in turn, is governed by
your experimental data. It may be inappropriate to start the axes at zero (e.g.
Fig. 49.1). In such instances, it is particularly important to show the scale
clearly, with scale breaks where necessary, so the graph does not mislead.
12
Note that Fig. 49.1 is drawn with ‘floating axes’ (i.e. the x and y axes do not
11 meet in the lower left-hand corner), while Fig. 49.2 has clear scale breaks on
both x and y axes.
10–3 3 number of animals
4
Graph paper
In addition to conventional linear (squared) graph paper, you may need the
3
following:
2 ●● Probability graph paper. This is useful when one axis is a probability scale
1
(e.g. p. 494).
●● Log-linear graph paper. This is appropriate when one of the scales shows a
0
logarithmic progression, e.g. in chemical kinetics. Log-linear paper is defined
0 10 12 14 16 18 20 22
Mass (g) by the number of logarithmic divisions (usually termed ‘cycles’) covered
(e.g. Fig. 49.4), so make sure you use a paper with the appropriate number
Fig. 49.3 Frequency distribution (histogram)
of masses for a sample of animals (sample
size 24 085); the size class interval is 2 g.
10000
Example For a data set where the

smallest number on the log axis is 1000
12 and the largest number is 9000,
three-cycle log-linear paper would be
used, covering the range 10–10 000.
100
Choosing between a histogram and a

bar chart – use a histogram for con-
10
tinuous quantitative variables and a 0 2 4 6 8 10
bar chart for discrete variables (see
Chapter 5 for details of these types of Fig. 49.4 Representation of three-cycle log-linear graph paper, marked up
measurement scale). to show a y axis (log) scale from 10 to 10 000 and an x axis (linear) scale
from 0 to 10.

Using graphs
20 of cycles for your data. An alternative approach is to plot the log-transformed

values on ‘normal’ graph paper.
15 ●● Log-log graph paper. This is appropriate when both scales show a loga-
rithmic progression.
Frequency
10
Types of graph
5
Different graphical forms may be used for different purposes.
0 ●● Plotted curves – used for data where the relationship between two variables
A B AB O
Blood group can be represented as a continuum (e.g. Fig. 49.1).
Fig. 49.5 Bar chart, showing the number ●● Scatter diagrams – used to visualise the relationship between individual
of students belonging to each ABO blood data values for two interdependent variables (e.g. Fig. 53.7) often as a pre-
group (n = 29). liminary part of a correlation analysis (p. 499).
●● Three-dimensional graphs show the interrelationships of three variables,
AB often one dependent and two independent (e.g. Fig. 55.5). A contour diagram
B is an alternative method of representing such data.
A
●● Histograms represent frequency distributions of continuous variables (e.g.
Fig. 49.3).
●● Frequency polygons emphasise the form of a frequency distribution by
joining the coordinates with straight lines, in contrast to a histogram. This
is particularly useful when plotting two or more frequency distributions on
the same graph.
●● Bar charts represent frequency distributions of a discrete qualitative or
O quantitative variable (e.g. Fig. 49.5). An alternative representation is the
line chart (e.g. Fig. 53.3).
Fig. 49.6 Pie chart: relative abundance of
human blood groups. ●● Pie charts illustrate portions of a whole (e.g. Fig. 49.6).
The plotted curve

Choosing graphical symbols –
plotted curves are usually drawn This is the commonest form of graphical representation used in biology. The
using a standard set of symbols: key features are outlined below and in checklist form in Box 49.1, while
● , ~ , ■, □, ▲, △, ◆, ◇. By convention, Box 49.2 advises on using Microsoft Excel.
paired symbols (‘closed’ and ‘open’) are
often used to represent ‘plus’ (treat- Data points
ment) and ‘minus’ (control) treatments. Each data point must be shown accurately, so that any reader can determine
the exact values of x and y. The results of each treatment must be readily
identifiable. A useful technique is to use a dot for each data point, surrounded
by a hollow symbol for each treatment (see Fig. 54.5) or to use symbols only
(Fig. 49.1–49.2), though the coordinates of each point are defined less accu-
rately. Use the same symbol for the same entity if it occurs in several graphs
and provide a key to all symbols.
Statistical measures
If you are plotting average values for several replicates and if you have the
necessary statistical knowledge, you can calculate the standard error (p. 486)
or the 95% confidence limits (p. 499) for each mean value and show these on
your graph as a series of vertical bars (see Fig. 49.1). Make it clear in the legend
whether the bars refer to standard errors or 95% confidence limits and quote

Using graphs
Box 49.1 Checklist for the stages in drawing a graph
The following sequence can be used whenever you need 6. Decide on appropriate descriptive labels for both axes,
to construct a plotted curve: it will need to be modified with SI units of measurement, where appropriate.
for other types of graph.
7. Choose the symbols for each set of data points and
1. Collect all of the data values and statistical values decide on the best means of representation for statis-
(in tabular form, where appropriate). tical values.
2. Decide on the most suitable form of presentation: 8. Plot the points to show the coordinates of each value
this may include transformation to convert data to with appropriate symbols.
linear form.
9. Draw a trend line for each set of points. Use a trans-
3. Choose a concise descriptive title, together with a ref- parent ruler, so you can draw the line to have an
erence (figure) number and date, where necessary. equal number of points on either side of it.
4. Determine which variable is to be plotted on the 10. Write a figure legend, to include a key which identifies
x axis and which on the y axis. all symbols and statistical values and any descriptive
footnotes.
5. Select appropriate scales for both axes and make
sure that the numbers and their location (scale marks)
are clearly shown, together with any scale breaks.
Box 49.2 H
ow to create and amend graphs within a spreadsheet (Microsoft Excel)
for use in coursework reports and dissertations
Microsoft Excel can be used to create graphs of reasona- Markers). Once selected, this will produce an embed-
ble quality, as long as you know how to amend the default ded scatter chart of the type shown in Fig. 49.7(a). The
settings so that your graph meets the formal standards line is then added later, as described below.
required for practical and project reports. As with a
hand-drawn graph, the basic stages in graph drawing 2. Change the default settings to improve the appear-
(Box 49.1) still apply. The following instructions explain ance of your graph. Consider each element of the
how to produce an X–Y graph (plotted curve, p. 453), image in turn, including the overall size, height and
bar graph (p. 453), pie graph and histogram using Excel, width of the graph (resize by clicking and dragging
where all types of graphs are termed ‘charts’. Earlier ver- one of the ‘sizing handles’ around the edge of the
sions use broadly similar commands, though not always chart). The graph shown in Fig. 49.7(b) was produced
in the same locations within the software. by altering the default settings, typically by moving
the cursor over the feature and then clicking the right
Producing an X-Y graph (Scatter chart in Excel) mouse button to reveal an additional menu of editing
and formatting options. (Note that the example given
1. Create the appropriate type of graph for your data.
below is for illustrative purposes only, and should not
Enter the numeric values for your X variable data in the
necessarily be regarded as prescriptive.)
cells of a single column and the equivalent values for
the Y variable in the adjacent cells of the next column Example for an X–Y graph (compare Fig. 49.7(a) with
to the right. Then select the whole data array (highlight Fig. 49.7(b)):
the appropriate cells by clicking and holding down the
left mouse button and dragging the cursor across ●● Unnecessary legend box on right-hand side can be
the cells so that all values are included). Then select removed using the (right-click) Delete option.
the Insert tab at the top of the sheet, and select (left-
●● Border to chart can be removed using Format Chart
click) the Scatter chart from the options provided in the
Area function (available by right-clicking within chart
upper ribbon. Note that you should never use the Line
area).
chart option, as it is based on an x axis that is not quan-
titative, so all X values will appear as equally spaced ●● Gridlines can either be removed, using the Delete func-
categories, rather than having a true scale). Select the tion or, if desired (as in Fig. 49.7(b), changed by click-
first option from the Scatter menu (Scatter with only ing on each axis and using the Add Minor Gridlines

Using graphs
(a) Producing a bar graph (Column chart in Excel)

1.8 1. Create the appropriate type of graph for your data.
1.6 Enter the category names (for x axis) in one column
1.4 and the numeric values (for y axis) in the next col-
1.2 umn. Select (highlight) the data array, then select
1.0 Series1 the Insert tab, and choose the Column chart from the
0.8 options provided. For a standard bar graph, select the
0.6
first option from the Column menu (Clustered Col-
0.4
umn). Once selected, this will produce an embedded
0.2
bar graph of the type shown in Fig. 49.8(a).
0.0
0 20 40 60 80 100 2. Change the default settings to improve the appear-
ance of your graph. The bar graph shown in
(b) Fig. 49.8(b) was produced by selecting each feature
2.0 and altering the default settings, as detailed below
(illustrative example).
Absorbance at 260 nm
1.5
Example for a bar graph (compare Fig. 49.8(a) with
Fig. 49.8(b)):
1.0
●● Unnecessary legend box on right-hand side can be
0.5 removed using the Delete option.
●● Border to chart can be removed using Format Chart
0.0
0 20 40 60 80 Area function available by right-clicking within chart
PAH concentration (µg mL–1) area.
Fig. 49.7 Examples of a plotted curve produced in Micro-

soft Excel using (a) default settings and (b) modified (a)
(improved) settings. 6
and Format Gridlines options to alter the Colour and 4

Style of the gridlines to make them more like those of Series1
3
conventional graph paper.
2
●● x and y axes can be reformatted by selecting each in
turn, and using the Format Axis menu options to select 1
appropriate scales for major and minor units, style of
0
tick marks, etc. Remember that it is better to use a fig- A B C
ure legend in Word (Chapter 45), rather than the Chart
Title option within Excel. (b)
●● x and y axis labels can be added by selecting the Lay- 6 5.7
out tab, then Axis Titles, then Primary Horizontal Axis
5
Title and Primary Vertical Axis Title options, which will
Mean weight (g)
produce a text box beside each axis into which can be 4 3.7
typed the axis label and any corresponding units. This 3
can then be changed from the default font using the 2.1
Home tab options. 2
1
●● Data point style can be changed by selecting
(right-clicking) any data point and following the For- 0
A B C
mat Data Series options to choose appropriate styles,
Treatment
colours and fill of the data markers.
●● A straight line of best fit can be added by selecting Fig. 49.8 Examples of a bar graph produced in Micro-
any data point and using the Add Trendline option to soft Excel using (a) default settings and (b) modified
choose a Linear line type with appropriate colour and (improved) settings.
style (explore other options within the Format, Layout
and Design tabs at the top of the worksheet). (continued)

Using graphs
●● Gridlines can either be removed, using the Delete

function or changed by selecting the gridlines and
using the Format Gridlines option to alter the Colour 21 31 Roots
and Style. Stem and petioles
Leaves
●● The y axis can be reformatted by selecting the axis, 5
then using the Format Axis menu options to select Flowers
appropriate scales, tick marks, line colour, etc. Note 19 24 Fruits
that the x axis should already contain category labels
from the spreadsheet cells (modify original cells to
update spreadsheet, if necessary).
Fig. 49.9 Example of a pie graph produced in Microsoft
●● x and y axis labels can be added by selecting the Lay- Excel using format modified from the default settings.
out tab, then Axis Titles, then Primary Horizontal Axis
Title and Primary Vertical Axis Title options, as detailed
for the plotted curve (Fig. 49.7). Producing a histogram
The histogram function in Excel requires a little more
●● Bar colour can be modified using the Format Data
effort to master, compared with other chart types. Essen-
Series menu, selecting appropriate Fill and Border col-
tially, a histogram is a graphical display of frequencies
ours, e.g. white and black, respectively, in Fig. 49.8(b).
(counts) for a continuous quantitative variable (p. 6),
●● Individual Y data values can be shown using the Add where the data values are grouped into classes. It is pos-
Data Labels option (other options and adjustments can sible to select the upper limit for each class into which
be made using the Format, Layout and Design tabs at you want the data to be grouped (these are termed ‘bin
the top of the worksheet). range values’ in Excel). An alternative approach is to
let the software select the class intervals (bin range
Note that for all types of graph, it is better not to
values) for you: Excel selects evenly distributed bins
use the Chart Title option within Excel, which places
between the minimum and maximum values. However,
the title at the top of the chart (as in Fig. 49.8(a)), but
this is often less effective than selecting your own class
to copy and paste your untitled graph into a word-
intervals.
processed document, such as a Microsoft Word file and
To use the Excel histogram function, you will need
then type a formal figure legend below the graph, as in
to make sure that the Analysis ToolPak is loaded
Fig. 49.7(b) and Fig. 49.8(b). However, once your graph
(check under the Data tab and, if not loaded, select via
is embedded into a Word file, it is generally best not to
the Office Button 7 Excel Options 7 Add@ins 7 Manage
make further amendments – you should go back to the
Excel Add@ins 7 Analysis ToolPak. The Histogram func-
original Excel file, make the required changes and then
tion should then be available in the Analysis ToolPak
repeat the copy-paste procedure to reinsert the graph
option on the Data tab/ribbon.
into the Word file.
The following steps outline the procedure used to cre-
ate the histograms shown in Fig. 49.10 for the table of
Producing a pie graph (Pie chart in Excel) data below (concentration of benzo (a) pyrene, in mg kg -1
in 24 soil samples).
1. Create the appropriate type of graph for your data.
Enter the category names for each part of the pie 7.2 6.5 7.1 8.5 6.6 7.2
chart in one column, and the corresponding numbers
(counts, fractions or percentages) in the next column. 7.0 7.3 8.6 9.1 7.5 8.3
Select (highlight) the data array, then select (left- 7.1 5.7 7.3 7.6 6.9 7.1
click) the Insert tab, and choose the Pie chart from
the options provided. For a standard pie graph, select 8.3 7.6 5.4 8.6 7.9 8.0
the first option from the menu (Pie). Once selected,
this will produce an embedded pie graph. 1. Enter the raw data values, e.g. as a single column of
numbers, or as an array, as above.
2. Change the default settings to improve the appear-
ance of your graph. For example, you can show the 2. Decide on the class intervals to be used. Base
data values (Data labels), adjust colours and shad- your choice on the number of data points and the
ing (e.g. switch from multi-colour to shades of grey), maximum and minimum values (for a small data
remove chart border, etc., as required (an illustrative set such as that shown above, you can do this by
example is shown as Fig. 49.9). visual examination, whereas for a large data set,

Using graphs
use the Excel functions COUNT, MAX and MIN (find (a)
these under the Σ symbol (More Functions) on the
Histogram
Editing section on the Home tab/ribbon, or use the 14
Descriptive Statistics 7 Summary Statistics option 12
of the Data Analysis component on the Data tab/rib- 10
bon). A typical histogram would have 4–10 classes,
Frequency
8
depending on the level of discrimination required. Frequency
6
Enter the upper limit for each class (Bin Range Val-
4
ues) in ascending order in a separate array of cells,
2
e.g. in a column close to the data values (in the
0
above example, 6, 7, 8 and 9 were chosen – the few 6 7 8 9 More
data values above the final bin value will be shown Bin
on the histogram as a group labelled ‘more’).
(b)
3. Select the histogram function, then input your data
and class interval values. From the Data tab/ribbon, 15
select Data Analysis 7 Histogram. A new window

will open: input your data into the Input Range box
10
(highlight the appropriate cells by clicking on the
Frequency
first data value and dragging to the final data value
while holding down the left mouse button). Next, 5
input the Bin Range values into the appropriate box
(if this is left empty, Excel will select default bin
range values). Most of the remaining boxes can 0
#6 6.1–7.0 7.1–8.0 8.1–9.0 .9.0
be left empty, though you must click the last box
Length (mm)
to get a Chart Output, otherwise the software will
give the numerical counts for each group, with-
Fig. 49.10 Examples of histogram output from Micro-
out drawing a histogram. Click OK and entries will
soft Excel using (a) default settings and (b) modified
be created within a new worksheet, showing the
(improved) settings.
upper limits of each group (in a column labelled
Frequency), plus a poorly constructed chart based
on Excel default settings, as shown in Fig. 49.10(a) ●● Bar chart converted to correct histogram format
(note that the default output is a bar chart, rather (no gaps between bars) using the Format Data
than a histogram, since there are gaps between the Series 7 Series Options, setting Gap Width to 0%.
groups).
●● Figure legend can be added below figure in Microsoft
Word, following copying and pasting of the Excel his-
Example for a histogram (compare Fig. 49.10(a) with togram into a Word file.
Fig. 49.10(b)):
Importing an Excel chart into a Word document
●● Chart can be resized to increase height, using the ‘siz-
ing handles’ at the edges of the chart and border line One problem encountered with current Microsoft Office
around graph can be removed using Format Chart products (but not with earlier versions, e.g. Office 1997–
Area 7 Border Color 7 No line (menu available by 2003) is that the standard Cut 7 Paste procedure gives
right-clicking within chart area). a poor quality figure, with grainy appearance and fuzzy
lines/text; similar problems occur using the Insert tab/
●● Title and unnecessary legend box can be removed
ribbon in Word. The simplest approach is to follow the
using the Delete option.
step-wise procedure below:
●● Axis scales can be reformatted using the Format Axis
1. Select your Excel chart: right-click outside the chart
menu options (e.g. scales, tick marks, line colour).
itself, near to the edge, then choose the Copy option
●● The x axis labels (class intervals) can be amended by from the drop-down menu.
typing directly into the cells containing the bin range
values. 2. Open your Word file, go to the Home tab/ribbon and
select Paste Special 7 Microsoft Excel Chart from the
●● Axis titles can be changed by typing directly into the options.
axis title box (double-click to access).
3. This should give a graph with the same crisp axis/
●● Bar colour can be changed (e.g. to grey, with black outline/text formatting as the original chart in Excel.
line) using the Format Data Series 7 Fill and Border
Colour options. (continued)

Using graphs
The alternative approach is to use Excel to print the entire Note: instructions illustrated here may vary among the
graph (chart) as a single sheet, and then add this to the print- different versions of Microsoft Office programmes. Check
out from your word-processed document. However, the dis- the exact functions and syntax using the ‘Insert function’
advantage with this approach is that you cannot produce a menu option (Fx button in some versions).
professional looking figure legend below your graph.
the value of n (the number of replicates per data point). Another approach is to
Adding error bars to Microsoft Excel
add a least significant difference bar (p. 498) to the graph.
graphs – you can do this as follows
(for Excel):
Interpolation
1. On your graph, right-click on one Once you have plotted each point, you must decide whether to link them by
of the data points of the series to straight lines or a smoothed curve. Each of these techniques conveys a dif-
which you want to add error bars; ferent message to your reader. Joining the points by straight lines may seem
2. From the Chart Tools 7 Layout tab, the simplest option, but may give the impression that errors are very low or
then Analysis group, select Error non-existent and that the relationship between the variables is complex. Join-
Bars and then choose the appro- ing points by straight lines is appropriate in certain graphs involving time
priate option from the menu. Typ- sequences (e.g. the number of animals at a particular site each year), or for
ically, you will wish to select More repeat measurements where measurement error can be assumed to be minimal
Error Bars Options 7 Custom and (e.g. recording a patient’s temperature in a hospital, to emphasise any variation
choose appropriate options from from one time point to the next). However, in most plotted curves the best
that menu. You may need to select straight line or curved line should be drawn (according to appropriate mathe-
a data array into which you have matical or statistical models, or by eye) to highlight the relationship between
entered calculated values for the the variables – after all, your choice of a plotted curve implies that such a rela-
upper and lower error limits for tionship exists. Do not worry if some of your points do not lie on the line: this
your chosen statistic over the range is caused by errors of measurement and by biological variation. Most curves
of the plotted values. You can adjust drawn by eye should have an equal number of points lying on either side of the
the format of the error bars from line. You may be guided by 95% confidence limits, in which case your curve
the More Error Bars Options menu. should pass within these limits wherever possible.
Curved lines can be drawn using a flexible curve, a set of French curves or
freehand. In the latter case, turn your paper so that you can draw the curve in a
single, sweeping stroke by a pivoting movement at the elbow (for larger curves)
Conveyìng the correct message – the or wrist (for smaller ones). Do not try to force your hand to make complex,
golden rule is: ‘always draw the sim- unnatural movements as the resulting line will not be smooth.
plest line that fits the data reasonably
well and is biologically reasonable’. Extrapolation
Be wary of extrapolation beyond the upper or lower limit of your measured val-
ues. This is rarely justifiable and may lead to serious errors. Whenever extrap-
Extrapolating plotted curves – try to olation is used, a dotted line ensures that the reader is aware of the uncertainty
avoid the need to extrapolate by better involved. Any assumptions behind an extrapolated curve should also be stated
experimental design. clearly in your text.
The histogram
While a plotted curve assumes a continuous relationship between the variables
by interpolating between individual data points, a histogram involves no such
assumptions. Histograms are also used to represent frequency distributions
(p. 452), where the y axis shows the number of times a particular value of
x was obtained (e.g. Fig. 49.2). As in a plotted curve, the x axis represents a

Using graphs
continuous quantitative variable which can take any value within a given range
Drawing a histogram – each datum is
(e.g. absorbance), so the scale must be broken down into discrete classes and
represented by a column with an area
the scale marks on the x axis should show either the mid-points (mid-values)
proportional to the magnitude of y: in
of each class, or the boundaries between the classes.
most cases, you should use columns of
The columns are contiguous (adjacent to each other) in a histogram, in
equal width, so that the height of each
contrast to a bar chart, where the columns are separate because the x axis of a
column is then directly proportional to y.
bar chart represents discrete values.
Shading or stippling may be used to
identify individual columns, according
to your needs. Interpreting graphs
The process of analysing a graph can be split into five phases:
1. Consider the context. Look at the graph in relation to the aims of the
Using computers to produce
study in which it was reported. Why were the observations made? What
graphs – never allow a computer pro-
hypothesis was the experiment set up to test? This information can usu-
gram to dictate size, shape and other
ally be found in the Introduction or Results section of a report. Also
aspects of a graph: find out how to alter
relevant are the general methods used to obtain the results. This might
scales, labels, axes, etc. and make appro-
be obvious from the figure title and legend, or from the Materials and
priate selections (see Box 49.2 for Micro-
Methods section.
soft Excel). Draw curves freehand if the
program only has the capacity to join the 2. Recognise the graph form and examine the axes. First, what kind of
individual points by straight lines. graph is presented (e.g. histogram, plotted curve)? You should be able to
recognise the main types summarised on p. 471 and their uses. Next, what
do the axes measure? You should check what quantity has been measured
in each case and what units are used.
Interpreting proportions and percentages
– you will need to establish how the 3. Look closely at the scale of each axis. What is the starting point and what
values have been calculated (see p. 85). is the highest value measured? For the x axis, this will let you know the
scope of the treatments or observations (e.g. whether they lasted for 5 min
or 20 years; whether a concentration span was 2-fold or 50-fold). For each
Examining graphs – do not be tempted axis, it is especially important to note whether the values start at zero; if
to look at the data displayed within a not, then the differences between any treatments shown may be magnified
graph before you have considered its by the scale chosen (see Box 49.3).
context, read the legend and the scale 4. Examine the symbols and curves. Information will be provided in
of each axis. the key or legend to allow you to determine to what these refer. If you
have made your own photocopy of the figure, it may be appropriate
to note this directly on it. You can now assess what appears to have
Understanding graphs within scientific happened. If, say, two conditions have been observed while a variable
papers – the legend should be a suc- is altered, when exactly do they differ from each other; by how much;
cinct summary of the key information and for how long?
required to interpret the figure without 5. Evaluate errors and statistics. It is important to take account of variabil-
further reference to the main text. This ity in the data. For example, if mean values are presented, the underlying
is a useful approach when ‘skimming’ a errors may be large, meaning that any difference between two treatments
paper for relevant information (p. 545). or observations at a given x value could simply have arisen by chance.
Thinking about the descriptive statistics used (Chapter 52) will allow you
to determine whether apparent differences could be significant in both
statistical and biological senses.
Sometimes graphs are used to mislead. This may be unwitting, as in an
unconscious favouring of a ‘pet’ hypothesis of the author. Graphs may be used
to ‘sell’ a product in the field of advertising or to favour a viewpoint as, per-
haps, in politics. Experience in drawing and interpreting graphs will help you
spot these flawed presentations, and understanding how graphs can be errone-
ously presented (Box 49.3) will help you avoid the same pitfalls.

Using graphs
Box 49.3 How graphs can misrepresent and mislead
1. The ‘volume’ or ‘area’ deception – this is mainly (a) (b)

found in histogram or bar chart presentations
where the size of a symbol is used to repre-
sent the measured variable. For example, the
amount of hazardous waste produced in differ-
ent years might be represented on a chart by dif-
ferent sizes of a chemical drum, with the y axis
(height of drum) representing the amount of
1972 2002 1972 2002
waste. However, if the symbol retains its shape
for all heights, as in Fig. 49.11(a), its volume will Fig. 49.11 Increase in pesticide use over a 30-year period.
increase as a cubic function of the height, rather
than in direct proportion. To the casual observer,
a twofold increase may look like an eightfold
one, and so on. Strictly, the height of the sym-
bol should be the measure used to represent the
(a) 200 (b) 250
variable, with no change in symbol width, as in
Fig. 49.11(b). 199 200
Mass (g) 198 150
Mass (g)
2. Effects of a non-zero axis – a non-zero axis acts to
emphasise the differences between measures by 197 100
reducing the range of values covered by the axis. 196 50
For example, in Fig. 49.12(a), it looks as if there
are large differences in mass between males and 195 0
Male Female Male Female
females; however, if the scale is adjusted to run
from zero (Fig. 49.12(b)), then it can be seen that Fig. 49.12 Average mass of males and females in test group.
the differences are not large as a proportion of
the overall mass. Always scrutinise the scale val-
ues carefully when interpreting any graph.
3. Use of a relative rather than absolute scale – this is

(a) 100 (b) Absolute response(measured units) 300
similar to the above, in that data compared using
relative scales (e.g. percentage or ratio) can give 250
Relative response (%)
80
the wrong impression if the denominator is not 60
200
the same in all cases. In Fig. 49.13(a), two treat- 150
ments are shown as equal in relative effect, both 40
100
resulting in 50% relative response compared (say) 20 50
to the respective controls. However, if treatment
0 0
A is 50% of a control value of 200 and treatment Treatment A Treatment B Treatment A Treatment B
B is 50% of a control value of 500, then the actual
difference in absolute response would have been Fig. 49.13 Responses to treatments A and B.
masked, as shown by Fig. 49.13(b).
4. Effects of a non-linear scale – when interpreting

graphs with non-linear (e.g. logarithmic) scales,
you may interpret any changes on an imagined
(a) 6 (b)
linear scale. For example, the pH scale is loga-
Cell density (cells per mL)
rithmic, and linear changes on this scale mean 200 000

log cell density
less in terms of absolute H + concentration at

high (alkaline) pH than they do at low (acidic) 4
100 000
pH. In Fig. 49.14(a), the cell density in two media
is compared on a logarithmic scale, while in
Fig. 49.14(b), the same data are graphed on a
2 0
linear scale. Note, also, that the log y axis scale Medium A Medium B Medium A Medium B
in Fig. 49.14(a) cannot be shown to zero, because
there is no logarithm for 0. Fig. 49.14 Effect of different media on cell density.

Using graphs
5. Unwarranted extrapolation – a graph may be (a) (b)
extrapolated to indicate what would happen if a

trend continued, as in Fig. 49.15(a). However, this
can only be done under certain assumptions (e.g.
that certain factors will remain constant or that rela-
tionships will hold under new conditions). There
may be no guarantee that this will actually be the
case. Fig. 49.15(b) illustrates other possible out-
comes if the experiment were to be repeated with Fig. 49.15 Extrapolation of data under different
higher values for the x axis. assumptions.
6. Failure to account for data point error – this mis-

representation involves curves that are overly
complex in relation to the scatter in the under-
lying data. When interpreting graphs with com-
plex curves, consider the errors involved in the (a) (b)
data values. It is probably unlikely that the curve
would pass through all the data points unless the
errors were very small. Fig. 49.16(a) illustrates a
curve that appears to assume zero error and is
thus overly complex, while Fig. 49.16(b) shows a
curve that takes possible errors of the points into
account.
Fig. 49.16 Fitted curves under different assumptions of
7. Failure to reject outlying points – this is a special
data error.
case of the previous example. There may be many
reasons for outlying data, from genuine mistakes to
statistical ‘freaks’. If a curve is drawn through such
points on a graph, it indicates that the point carries
equal weight with the other points, when in fact,
it should probably be ignored. To assess this, con-
(a) (b)
sider the accuracy of the measurement, the num-
ber and position of adjacent points and any special
factors that might be involved on a one-off basis.
Fig. 49.17(a) shows a curve where an outlier has
perhaps been given undue weight when showing
the presumed relationship. If there is good reason
to think that the point should be ignored, then the
curve shown in Fig. 49.17(b) would probably be
more valid. Fig. 49.17 Curves with and without outlier taken into
account.
8. Inappropriate fitted line – here, the mathematical
function chosen to represent a trend in the data
might be inappropriate. A straight line might be
fitted to the data, when a curve would be more cor-
rect, or vice versa. These cases can be difficult to
assess. You need to consider the theoretical valid- (a) (b)
ity of the model used to generate the curve (this is

not always stated clearly). For example, if a straight
line is fitted to the points, the implicit underlying
model states that one factor varies in direct relation
to another, when the true situation may be more
complex. In Fig. 49.18(a), the relationship has been
shown as a linear relationship, whereas an expo-
nential relationship, as shown in Fig. 49.18(b), could Fig. 49.18 Different mathematical model used to repre-
be more correct. sent trends in data.

Using graphs

Billo, E.J. (2011) Excel for Chemists: A comprehensive Miller, J.N. and Miller, J.C. (2010) Statistics and Chemo
guide, 3rd edn. John Wiley & Sons Ltd, Chichester. metrics for Analytical Chemistry, 6th edn. Prentice Hall,
Briscoe, M.H. (1996) Preparing Scientific Illustrations: A Harlow.
Guide to Better Posters, Presentations and Publications. Monk, P. and Munro, L.J. (2010) Maths for Chemists: A
Springer-Verlag, New York. Chemist’s Toolkit of Calculations. Oxford University
Currell, G. and Dowman, T. (2009) Essential Mathematics Press, Oxford.
and Statistics for Science, 2nd edn. John Wiley & Sons Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
Ltd, Chichester. Oxford University Press, Oxford.
De Levie, R. (2001) How to use Excel in Analytical Chem
istry and in General Scientific Data Analysis. Cambridge
University Press, Cambridge.
Study Exercises
49.1 Select appropriate graphical presenta- 49.3 Create a frequency distribution histogram. The
tions. Choose an appropriate graphical form for table below gives data for the fructose concen-
each of the following examples: tration in 100 samples of fruit juice. Plot a histo-
gram showing the frequency distribution of the
(a) A titration curve showing the influence of
data. Write a brief description of the important
0.1 M NaOH on pH following the addition of
features of the distribution.
0.1 M HCl.
(b) The concentration of lead in blood from 100 Fructose content (g L-1) in fruit juice
children.
11.1 14.2 13.5 9.8 12.0 13.9 14.1 14.6 11.0 12.3
(c) The determination of selenium in bread flour.
(d) The sand, silt and clay content of a soil 13.4 12.9 12.9 10.0 13.1 11.8 12.6 10.7 8.1 11.2
sample. 13.8 12.4 12.9 11.3 12.7 12.4 14.6 15.1 11.2 9.7
(e) A quality control chart for the paracetamol
11.3 14.7 10.8 13.3 11.9 11.4 12.5 13.0 11.6 13.1
content of tablets.
9.3 13.5 14.6 11.2 11.7 10.9 12.4 12.0 12.1 12.6
49.2 Create a pie chart. Display the following infor-
10.9 12.1 13.4 9.5 12.5 11.6 12.2 8.8 10.7 11.1
mation in the form of a pie chart.
10.2 11.7 10.4 14.0 14.9 11.5 12.0 13.2 12.1 13.3
Typical composition of a breakfast cereal.
12.4 9.4 13.2 12.5 10.8 11.7 12.7 14.1 10.4 10.5
Content Typical value per 100 g 13.3 10.6 10.5 13.7 11.8 14.1 10.3 13.6 10.4 13.9
Protein 6g 11.7 12.8 10.4 11.9 11.4 10.6 12.7 11.4 12.9 12.1
Carbohydrate 83 g
49.4 Find examples of misleading graphs. Create a port-
Fat 5g folio of examples of misleading graphs taken from
Fibre 4g newspapers. For each graph, state what aspect is
misleading (see Box 49.3) and, where possible,
Other 2g
attempt to show the data correctly in a new graph.

50 Presenting data in tables
A table is often the most appropriate way to present numerical data in a concise,
Alternatives to tables for presenting
accurate and structured form. Assignments and project reports should contain
numerical data – if you have only a
tables that have been designed to condense and display results in a meaningful
few numbers, consider simply present-
way and to aid numerical comparison. The preparation of tables for recording
ing these within the text; an alternative
primary data is discussed Chapter 3.
approach is to show the data values on
Decide whether you need a table, or whether a graph is more appropriate.
a bar chart (e.g. Fig. 49.5).
Histograms and plotted curves can be used to give a visual impression of the
relationships within your data (Chapter 49). On the other hand, a table gives
you the opportunity to make detailed numerical comparisons.
Key Point Always remember that the primary purpose of your

table is to communicate information and allow appropriate com-
parison, not simply to put down the results on paper.
Preparation of tables
Title
Every table must have a brief descriptive title. If several tables are used, number
Constructing titles – take care over
them consecutively so they can be quoted in your text. The titles within a report
titles as it is a common mistake in stu-
should be compared with one another, making sure they are logical and consist-
dent practical reports to present tables
ent and that they describe accurately the numerical data contained within them.
without titles, or to misconstruct the title.
Structure
Display the components of each table in a way that will help the reader under-
stand your data and grasp the significance of your results. Organise the columns
so that each category of like numbers or attributes is listed vertically, while each
Saving space in tables – you may
be able to omit a column of control
horizontal row shows a different experimental treatment, organism, sampling
data if your results can be expressed
site, etc. (as in Table 50.1). Where appropriate, put control values near the
as percentages of the corresponding
beginning of the table. Columns that need to be compared should be set out
control values.
alongside each other. Use rulings to subdivide your table appropriately, but
avoid cluttering it up with too many lines.
Table 50.1 Selected properties of elements in the Periodic Table
Atomic
Atomic Relative radius Important Density
Element Symbol number atomic mass** (pm) oxidation states (g mL -1)* Compounds in ores
Aluminium Al 13 26.98 143 +3 2.7 Al2O3

Cobalt Co 27 58.93 125 + 2, + 3 8.7 CoAsS, CoAs2, CoS
Iron Fe 26 55.84 126 + 2, + 3 7.9 Fe2O3, Fe3O4, FeCO3, FeS2
Manganese Mn 25 54.94 126 + 2, + 3, + 4, + 7 7.2 MnO2, Mn2O3, Mn3O4,
mixed oxide
Zinc Zn 30 65.38 138 +2 7.1 ZnS, ZnO, ZnCO3
*Determined at 28 °C.
**Relative to the atomic mass of 12C ( = 12).

Presenting data in tables
Headings and subheadings

These should identify each set of data and show the units of measurement,
where necessary. Make sure that each column is wide enough for the headings
and for the longest data value.
Numerical data
Example Quote the radius of Within the table, do not quote values to more significant figures than necessary,
an atom as 0.126 nm or 126 pm, as this will imply spurious accuracy (pp. 38). By careful choice of appropriate
rather than 0.000 000 000 126 m or units for each column you should aim to present numerical data within the
0.126 * 10-9 m (or 126 * 10-12 m). range 0 to 1000. As with graphs, it is less ambiguous to use derived SI units,
However, some texts still use the ang- with the appropriate prefixes, in the headings of columns and rows, rather than
strom, Å. An angstrom is equivalent to quoting multiplying factors as powers of 10. Alternatively, include exponents
10-10 m; hence the radius of an atom in the main body of the table (see Table 12.1) to avoid any possible confusion
would be 1.26 Å in this example.
regarding the use of negative powers of 10.
Other notations
Saving further space in tables – in Avoid using dashes in numerical tables, as their meaning is unclear; enter a zero
some instances a footnote can be reading as ‘0’ and use ‘NT’ not tested or ‘ND’ if no data value was obtained,
used to replace a whole column of with a footnote to explain each abbreviation. Other footnotes, identified by
repetitive data. asterisks, superscripts or other symbols in the table, may be used to provide
relevant experimental detail (if not given in the text) and an explanation of
column headings and individual results, where appropriate. Footnotes should
be as condensed as possible. Table 50.1 provides examples.
Statistics
In tables where the dispersion of each data set is shown by an appropriate statis-
tical parameter, you must state whether this is the (sample) standard deviation,
the standard error (of the mean) or the 95% confidence limits and you must give
the value of n (the number of replicates). Other descriptive statistics should be
quoted with similar detail, and hypothesis-testing statistics should be quoted
along with the value of P (the probability). Details of any test used should be
given in the legend, or in a footnote.
Text
Sometimes a table can be a useful way of presenting textual information in a
Using spreadsheets and word process-
condensed form (see example on pp. 312 and 463).
ing packages – these can be used to
When you have finished compiling your tabulated data, carefully
prepare high-quality versions of tables
double-check each numerical entry against the original information, to ensure
for project work (Box 50.2).
that the final version of your table is free from transcriptional errors. Box 50.1
gives a checklist for the major elements of constructing a table.
Box 50.1 Checklist for preparing a table
Every table should have the following components.

1. A title, plus a reference number and date where necessary.
2. Headings for each column and row, with appropriate units of
measurement.
3. Data values, quoted to the nearest significant figure and with statis-
tical parameters, according to your requirements.
4. Footnotes to explain abbreviations, modifications and individual
details.
5. Rulings to emphasise groupings and distinguish items from each other.

Box 50.2 H
ow to use a word processor (Microsoft Word) or a spreadsheet (Microsoft Excel)
to create a table for use in coursework reports and dissertations
Creating tables with Microsoft Word Heading 1 Heading 2 Heading 3 Heading 4

Word-processed tables are suitable for text-intensive or xx xx xx xx
number-intensive tables, although in the second case
entering data can be laborious. When working in this way, xx xx
the natural way to proceed is to create the ‘shell’ of the
table, add the data, and then carry out final formatting 6. Finally, remove selected borders to cells – one way
on the table. is to highlight the table, then click on the Design tab
1. Move the cursor to the desired position in your and choose an appropriate style from the template
document – this is where you expect the top left cor- options. If you do not want any shading, click on
ner of your table to appear. Click the Insert tab, then Design 7 Shading and choose No Color. Another way
choose Table. is to highlight the cells in the table, then right-click the
mouse button and choose Borders and Shading. You
2. Select the appropriate number of columns and can choose the style you wish from this submenu so
rows – do not forget to add rows and columns for that your table looks like the examples shown in this
headings. As default, a full-width table will appear, chapter.
with single rulings for all cell boundaries, with all
columns of equal width and all rows of equal height. 7. Add a table title – this should be positioned above the
table (cf. a figure title and legend, p. 490), legend and
Example of a 4 * 3 table:
footnotes.
Final version of the table:
Table xx. A table of some data
Heading 1 Heading 2a Heading 3 Heading 4
3. Customise the columns – by placing the cursor over xx xx xx xx

the vertical rulings then ‘dragging’, you can adjust
xx xx xx
their width to suit your heading text entries, which
should now be added. a
An example of a footnote.
Heading 1 Heading 2 Heading 3 Heading 4 Creating tables with Microsoft Excel

Tables derived from spreadsheets are effective when
you have lots of numerical data, especially when these
are stored or created using the spreadsheet itself. When
working in this way, you can design the table as part of
4. Work through the table adding the data – entries can an output or summary section of the spreadsheet, add
be numbers or text. explanatory headings, format, then possibly export to a
word processor when complete.
Heading 1 Heading 2 Heading 3 Heading 4
1. Design the output or summary section. Plan this as if
xx xx xx xx it were a table, including adding text headings within
xx xx xx cells.
5. Make further adjustments to column and row widths

to suit – for example, if text fills several rows within
a cell, consider increasing the column width, and if a
column contains only single or double digit numbers,
consider shrinking its width. To combine cells, first
highlight them, then right-click the mouse button (or
equivalent) and click on Merge Cells. You may wish 2. Insert appropriate formulae within cells to produce
to reposition text within a cell by right-clicking the data. If necessary, formulae should draw on the other
mouse button and choosing an appropriate position parts of the spreadsheet.
in the cell alignment menu. (continued)

3. Format the cells. This is important to con- 6. Add ‘real’ data values to the spreadsheet. This should
trol the number of decimal places presented result in the summary values within the table being
(Home 7 Number 7 Format 7 Cells). filled. Check that these are presented with the appro-
4. Adjust column width to suit. You can do this via the priate number of significant figures (pp. 41).
column headings, by placing the cursor over the rul- 7. The table can now be copied and pasted to a Word
ings between columns then ‘dragging’. document. For best results, use the Paste Spe-
cial . . . 7 Microsoft Office Excel Worksheet Object
option.
Note: Instructions illustrated here may vary among the

different versions of Microsoft Office programmes. Check
5. Add rulings as appropriate. One way is to use the the exact functions and syntax using the ‘Insert function’
Home 7 Font 7 Borders menu, having selected the menu option (fx button in some versions.
relevant cells.

Billo, E.J. (2007) Excel for Scientists and Engineers. John Miller, J.N. and Miller, J.C. (2010) Statistics and Chemo
Wiley & Sons Ltd, Chichester. metrics for Analytical Chemistry, 6th edn. Prentice Hall,
Currell, G. and Dowman, T. (2009) Essential Mathematics Harlow.
and Statistics for Science, 2nd edn. John Wiley & Sons Monk, P. and Munro, L.J. (2010) Maths for Chemists: A
Ltd, Chichester. Chemist’s Toolkit of Calculations, Oxford University
Kirkup, L. (1994) Experimental Methods: An Introduction Press, Oxford.
to the Analysis and Presentation of Data. John Wiley & Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
Sons Ltd, New York. Oxford University Press, Oxford.
Study exercises
50.1 Redesign a table of data. Using the following Alcohol* 86 mg 100 mL-1
example, redraft the table to improve layout and Caffeine – –
correct inconsistencies. Nicotine – –
Concentrations of drug and metabolites found in a *nicotine and caffeine were not quantified
blood sample
50.2 Devise a text-based table. After reading through
Diazepam 0.2 mg L-1 this chapter, and working from memory, draw up
Desmethyl diazepam 0.075 mg mL-1
a table listing the principal components of a typi-
Temazepam 0.1 cal table in the first column, and brief comments
mg L-1
on the major features of each component in the
Methadone 0.23 mg mL-1
second column.

51 Hints for solving numerical problems
Chemistry often requires a numerical or statistical approach. Not only is math-

Advice on dealing with numerical
ematical modelling an important aid to understanding, but computations are
procedures – see:
often needed to turn raw data into meaningful information or to compare them
●● Box 9.1: preparing solutions (p. 72); with other data sets. Moreover, calculations are part of laboratory routine, per-
●● Box 9.2: preparing solutions (p. 77); haps required for making up solutions of known concentration (see p. 71 and
●● Box 11.1: molar concentrations (p. 103); below) and calibration curves (see Box 30.2). In research, ‘trial’ calculations
●● Box 48.1: calibration curves (p. 446); can reveal what input data are required and where errors in their measurement
●● Box 30.1: stock solutions (p 226); might be amplified in the final result (see p. 486).
●● Box 30.2: calibration solutions (p. 227);
●● Box 30.3: standard additions (p. 229);
●● Box 30.5: dilution factor (p. 230); Key Point If you find numerical work difficult, practice at
●● Box 32.1: calibration solutions (p. 257); problem-solving is especially important.
●● Box 35.1: radioactivity (p. 314).
Practising problem-solving:
●● demystifies the procedures involved, which are normally just the elemen-
tary mathematical operations of addition, subtraction, multiplication and
division (Table 51.1);
●● allows you to gain confidence so that you do not become confused when
confronted with an unfamiliar or apparently complex form of problem;
●● helps you to recognise the various forms a problem can take as, for
instance, in the different forms of titrations (Chapter 25–28).
Table 51.1 Sets of numbers and operations.
Sets of numbers
Whole numbers: 0, 1, 2, 3, . . .
Natural numbers: 1, 2, 3, . . .
Integers: . . . - 3, - 2, - 1, 0, 1, 2, 3, . . .
Real numbers: integers and anything between (e.g. - 5, 4.376, 3/16, p, 25)
Prime numbers: subset of natural numbers divisible by 1 and themselves only (i.e. 2, 3, 5, 7, 11, 13, . . . )
Rational numbers: p/q where p (integer) and q (natural) have no common factor (e.g. 3/4)
Fractions: p/q where p is an integer and q is natural (e.g. - 6/8)
Irrational numbers: real numbers with no exact value (e.g. p)
Infinity: (symbol ∞ ) is larger than any number (technically not a number as it does not obey the laws of algebra)
Operations and symbols
Basic operators: + , - , * and , will not need explanation; however, / may substitute for , , * may substitute for * or this
operator may be omitted
Powers: an, i.e. ‘a to the power n’, means a multiplied by itself n times (e.g. a2 = a * a = ‘a squared’,
a3 = a * a * a = ‘a cubed’), n is said to be the index or exponent. Note a0 = 1 and a1 = a
Logarithms: the common logarithm (log) of any number * is the power to which 10 would have to be raised to give x
(i.e. the log of 100 is 2; 102 = 100); the antilog of x is 10 * . Note that there is no log for 0, so take this into
account when drawing log axes by breaking the axis. Natural or Napierian logarithms (In) use the base
e ( = 2.71828c) instead of 10
Reciprocals: the reciprocal of a real number a is 1/a(a ≠ 0)
Relational a 7 b means ‘a is greater (more positive) than b’, 6 means less than, … means less-than-or-equal-to and
operators: Ú means greater-than-or-equal-to
Proportionality: a ∝ b means ‘a is proportional to b’ (i.e. a = kb, where k is a constant). If a ∞1/b, a is inversely proportional
to b (a = k/b)
Sums: Σxi is shorthand for the sum of all x values from i = 0 to i = n (more correctly, the range of the sum is speci-
fied under the symbol)
Moduli: x signifies modulus of x, i.e. its absolute value (e.g. 4 = - 4 = 4)
Factorials: x! signifies factorial x, the product of all integers from 1 to x (e.g. 3! = 6). Note 0! = 1! = 1

Hints for solving numerical problems
Steps in tackling a numerical problem

Tracing errors in mathematical problems
– this is always easier when all the The step-by-step approach outlined below may not be the fastest method of
stages in a calculation are laid out arriving at an answer, but most mistakes occur where steps are missing, com-
clearly. bined or not made obvious, so a logical approach is often better. Error tracing
is distinctly easier when all stages in a calculation are laid out.
Have the right tools ready

Using a computer spreadsheet for Scientific calculators (p. 4) greatly simplify the numerical part of
numerical problems – this may be problem-solving. However, the seeming infallibility of the calculator may
very useful in repetitive work or for lead you to accept an absurd result which could have arisen because of faulty
‘what if?’ case studies (see Chapter 45) key-pressing or faulty logic. Make sure you know how to use all the features on
(Adams, 2003). your calculator, especially how the memory works, how to introduce a constant
multiplier or divider, and how to obtain an exponent (note that the ‘exp’ button
on most calculators gives you 10x, not 1x or y x; so 1 * 106 would be entered
as 1 exp 6 , not 10 exp 6).
Approach the problem thoughtfully

If the individual steps have been laid out on a worksheet, the ‘tactics’ will
Table 51.2 Simple algebra – rules for
already have been decided. It is more difficult when you have to adopt a strat-
manipulating. egy on your own, especially if the problem is presented as a story and it is not
obvious which equations or rules need to be applied.
If a = b + c, then b = a - c and c = a - b
●● Read the problem carefully as the text may give clues as to how it should
If a = b * c, then b = a , c and c = a , b be tackled. Be certain of what is required as an answer before starting.
If a = bc, then b = a1/c and c = log a , log b ●● Analyse what kind of problem it is, which effectively means deciding
a 1/n n
= 2a which equation(s) or approach will be applicable. If this is not obvious,
consider the dimensions/units of the information available and think how
a -n = 1 , an
they could be fitted to a relevant formula. In examinations, a favourite ploy
ab * ac = a(b + c) and ab , ac = a(b - c) of examiners is to present a problem such that the familiar form of an equa-
(ab)c = a(b * c) tion must be rearranged (see Table 51.2 and Box 51.1). If you are unsure
whether a recalled formula is correct, a dimensional analysis can help: write
a * b = antilog(log a + log b)
in all the units for the variables and make sure that they cancel out to give
the expected answer.
●● Check that you have, or can derive, all of the information required to
use your chosen equation(s). It is unusual but not unknown for examiners
to supply redundant information. So, if you decide not to use some of the
information given, be sure why you do not require it.
Presenting calculations in assessed

●● Decide on what format and in which units the answer should be pre-
work – always show the steps in your
sented. This is sometimes suggested to you. If the problem requires many
calculations, as most markers will only
changes in the prefixes to units, it is a good idea to convert all data to base
penalise a mistake once and part marks
SI units (multiplied by a power of 10) at the outset.
will be given if the remaining opera- ●● If a problem appears complex, break it down into component parts.
tions are performed correctly. This can
only be done if those operations are Present your answer clearly
visible.
The way you present your answer obviously needs to fit the individual problem.
Multiple examples are shown in Chapters 22–28 and 30 that illustrate several
important points. Guidelines for presenting an answer include the following:
(a) Make your assumptions explicit. Most mathematical models of biolog-
ical phenomena require that certain criteria are met before they can be

Box 51.1 Example of using the algebraic rules of Table 51.2
Problem: if a = (b - c) , (d + en), find e. 3. Subtract d from both sides; formula becomes:

b - c
1. Multiply both sides by (d + en); formula becomes: en = - d
a
a(d + en) = (b - c)
4. Raise each side to the power 1/n; formula becomes:
2. Divide both sides by a; formula becomes: 1/n
b - c
b - c e = e - df
d + en = a
a
legitimately applied (e.g. ‘assuming the tissue is homogeneous . . . ’), while

some approaches involve approximations that should be clearly stated (e.g.
‘to estimate the mouse’s skin area, its body was approximated to a cylinder
with radius x and height y. . .’).
(b) Explain your strategy for answering, perhaps giving the applicable for-
mula or definitions which suit the approach to be taken. Give details of
what the symbols mean (and their units) at this point.
(c) Rearrange the formula to the required form with the desired unknown
Units – never write any answer
on the left-hand side (see Table 51-2 as well as examples in Chapters
without its unit(s) unless it is truly
22–28 and 30).
dimensionless.
(d) Substitute the relevant values into the right-hand side of the formula,
using the units and prefixes as given (it may be convenient to convert
values to SI beforehand). Convert prefixes to appropriate powers of 10 as
soon as possible.
(e) Convert to the desired units step by step, i.e. taking each variable in turn.
(f) When you have the answer in the desired units, rewrite the left-hand
side and underline the answer, for emphasis. Make sure that the result
is presented to an appropriate number of significant figures (see below).
Check your answer
Having written out your answer, you should check it methodically, answering
the following questions:
●● Is the answer realistic? You should be alerted to an error if a number is
absurdly large or small. In repeated calculations, a result standing out from
others in the same series should be double-checked.
●● Do the units make sense and match up with the answer required? For
example, do not present a volume in units of m2.
●● Do you get the same answer if you recalculate in a different way? If
you have time, recalculate the answer using a different ‘route’, entering the
numbers into your calculator in a different form and/or carrying out the
operations in a different order.
Rounding off to a specific number of
significant figures – do not round off Rounding: decimal places and significant figures
numbers until you arrive at the final
answer or you will introduce ‘rounding’
In many instances, the answer you produce as a result of a calculation will
errors into the calculation.
include more figures than is justified by the accuracy and precision of the
original data. Sometimes you will be asked to produce an answer to a specified

number of decimal places or significant figures, and at other times you will be
Examples expected to decide for yourself what would be appropriate.
The number 4.123 correct to two
decimal places is 4.12
The number 4.126 correct to two Key Point Do not simply accept the numerical answer from
decimal places is 4.13 a calculator or spreadsheet, without considering whether you
The number 4.1251 correct to two need to modify this to give an appropriate number of significant
figures or decimal places.
The number 4.1250 correct to two
The number 4.1350 correct to two Rounding to n decimal places
The number 99.99 correct to one This is relatively easy to do.
decimal place is 100.0. 1. Look at the number to the right of the nth decimal place.
2. If this is less than 5, simply ‘cut off’ all numbers to the right of the nth
decimal place (i.e. round down).
3. If the number is greater than 5, ‘cut off’ all numbers to the right of the
nth decimal place and add one to the nth decimal place (i.e. round up).
Examples 4. If the number is 5, then look at further numbers to the right to deter-
The number of significant figures in mine whether to round up or not.
194 is three
The number of significant figures in 5. If the number is exactly 5 and there are no further numbers to the
2305 is four right, then round the nth digit to the nearest even number. Note: When
The number of significant figures in considering a large number of calculations, this procedure will not affect
0.003 482 is four the overall mean value. Some rounding systems do the opposite to this (i.e.
The number of significant figures in round to the nearest odd number), while others always round up where the
210 * 108 is 3 (21 * 109 would be two). number is exactly 5 (which will affect the mean). Take advice from your
tutor and stick to one system throughout a series of calculations.
Whenever you see any numbers quoted, you should assume that the last
Examples digit has been rounded. For example, in the number 22.4, the ‘.4’ is assumed to
The number of significant figures in be rounded and the calculated value may have been between 22.35 and 22.45.
3051.93 is six
To five significant figures, this number
Quoting to n significant figures
is 3051.9 The number of significant figures indicates the degree of approximation in the
To four significant figures, this number number. For most cases, it is given by counting all the figures except zeros
is 3052 that occur at the beginning or end of the number. Zeros within the number are
To three significant figures, this always counted as significant. The number of significant figures in a number
number is 3050 like 200 is ambiguous and could be one, two or three; if you wish to specify
To two significant figures, this number clearly, then quote as e.g. 2 * 102 (one significant figure), 2.0 * 102 (two
is 3100
significant figures), etc. to avoid spurious accuracy (p. 193). When quoting a
To one significant figure, this number
is 3000
number to a specified number of significant figures, use the same rules as for
3051.93 to the nearest 10 is 3050 rounding to a specified number of decimal places, but do not forget to keep
3051.93 to the nearest 100 is 3100 zeros before or after the decimal point. The same principle is used if you are
Note that in this last case you must asked to quote a number to the ‘nearest 10’, ‘nearest 100’, etc.
include the zeros before the decimal When deciding for yourself how many significant figures to use, adopt the
point to indicate the scale of the following rules of thumb:
number (even if the decimal point is
not shown). For a number less than ●● Always round after you have completed a calculation. Use all significant
1, the same would apply to the zeros figures available in the measured data during a calculation.
before the decimal point. For example, ●● If adding or subtracting with measured data, then quote the answer to
0.003 051 93 to three s ignificant figures
the number of decimal places in the data value with the least number
is 0.003 05. Alternatively, use scientific
notation (in this case, 3.05 * 10-3).
of decimal places (e.g. 32.1 - 45.67 + 35.6201 = 22.1, because 32.1 has
one decimal place).

●● If multiplying or dividing with measured data, keep as many significant

figures as are in the number with the least number of significant figures
(e.g. 34 901 , 3445 * 1.341 0344 = 13.59, because 3445 has four sig-
nificant figures).
●● For the purposes of significant figures, assume ‘constants’ have an
infinite number of significant figures (e.g. number of millimetres in a
metre).
Some reminders of basic mathematics

Errors in calculations sometimes appear because of faults in mathematics rather
than computational errors. For reference purposes, Tables 51.1 and 51.2 give
some basic mathematical principles that may be useful. Eason et al. (1992)
should be consulted for more advanced/specific needs.
Examples
Percentages and proportions
1/8 as a percentage is A percentage is just a fraction expressed in terms of hundredths, indicated by
1 , 8 * 100 = 100 , 8 = 12.5% putting the percentage sign (%) after the number of hundredths. So 35% simply
0.602 as a percentage is means 35 hundredths. To convert a fraction to a percentage, just multiply the
0.602 * 100 = 60.2%. fraction by 100. When the fraction is in decimal form, multiplying by 100 to
obtain a percentage is easily achieved just by moving the decimal point two
places to the right.
To convert a percentage to a fraction, just remember that, since a per-
Examples centage is a fraction multiplied by 100, the fraction is the percentage divided
190% as a decimal fraction is
by 100. For example: 42% = 42/100 = 0.42. In this example, since we are
190 , 100 = 1.9
dealing with a decimal fraction, the division by 100 is just a matter of moving
5/2 as a percentage is
5 , 2 * 100 = 250%. the decimal point two places to the left (42% could be written as 42.0%). Per-
centages greater than 100% represent fractions greater than 1. Percentages less
than 1 may cause confusion. For example, 0.5% means half of one per cent
(0.005) and must not be confused with 50% (which is the decimal fraction 0.5).
Example A population falls from To find a percentage of a given number, just express the percentage as a
4 million to 3.85 million. What is the decimal fraction and multiply the given number. For example: 35% of 500 is
percentage change? The decrease in given by 0.35 * 500 = 175. To find the percentage change in a quantity, work
numbers is 4 - 3.85 = - 0.15 million. out the difference ( =value ‘after’ – value ‘before’), and divide this difference
The fractional decrease is by the original value to give the fractional change, then multiply by 100.
- 0.15 , 4 = -0.0375 and we mul-
tiply by 100 to get the percentage Exponents
change = minus 3.75%. Exponential notation is an alternative way of expressing numbers in the form
an (‘a to the power n’), where a is multiplied by itself n times. The number a is
called the base and the number n the exponent (or power or index). The expo-
nent need not be a whole number, and it can be negative if the number being
Example 23 = 2 * 2 * 2 = 8.
expressed is less than 1. See Table 51.2 for other mathematical relationships
involving exponents.
Scientific notation
In scientific notation, also known as ‘standard form’, the base is 10 and the
exponent a whole number. To express numbers that are not whole powers of
10, the form c * 10n is used, where the coefficient c is normally between
1 and 10. Scientific notation is valuable when you are using very large numbers
and wish to avoid suggesting spurious accuracy. Thus if you write 123 000, this
may suggest that you know the number to {0.5, whereas 1.23 * 105 might
give a truer indication of measurement accuracy (i.e. implied to be {500 in
this case). Engineering notation is similar, but treats numbers as powers of 10 in

groups of 3, i.e. c * 100, 103, 106, 109, etc. This corresponds to the SI system
Example Avogadro’s number,
≈602 352 000 000 000 000 000 000,
of prefixes (p. 40).
is more conveniently expressed as A useful property of powers when expressed to the same base is that
6.023 52 * 1023. when multiplying two numbers together, you simply add the powers, while, if
dividing, you subtract the powers. Thus, suppose you counted 8 bacteria in a
known value of a 10-7 dilution, there would be 8 * 107 in the same volume of
Examples The logarithm to the undiluted solution; if you now dilute this 500-fold (5 * 102), then the number
base 10 (log10) of 1000 is 3, since present in the same volume would be 8/5 * 10(7-2) = 1.6 * 105 = 160 000.
103 = 1000. The logarithm to the
base e (loge or In) of 1000 is 6.907 755
Logarithms
(to six decimal places). When a number is expressed as a logarithm, this refers to the power n to which
the base number a must be raised to give that number. Any base could be used,
but the two most common are 10, when the power is referred to as log10 or
Examples (use to check the correct simply log, and the constant e (2.718 282), used for mathematical convenience
use of your own calculator) in certain situations, when the power is referred to as loge or ln (natural loga-
102 963 as a log (to base 10) = rithm). Note that: (a) logs need not be whole numbers; (b) there is no log value
5.012 681 (to six decimal places) for the number zero; and (c) log10 = 0 for the number 1.
105.012681 = 1029 62.96 To obtain logs, you will need to use the log key on your calculator, or spe-
(Note loss of accuracy owing to cial log tables (now largely redundant). To convert back (‘antilog’) use
loss of decimal places.) 102 963 as a
natural logarithm (In) = 11.542 125 ●● the 10x key, with x = log value;
(to six decimal places) thus ●● the inverse then the log key; or
2.718 28211.542125 = 102 963.
●● the y x key, with y = 10 and x = log value.
If you have used log tables, you will find complementary antilogarithm tables
Example Using Eqn [51.3], the pre- to do this.
dicted value for y for a linear function
There are many uses of logarithms in chemistry, including pH (= -log[H + ]),
where a = 2 and b = 0.5, where x = 8
is: y = 2 + (0.5 * 8) = 6.
where [H + ] is expressed in m o l L-1 (see p. 113); rate constants in physical
chemistry, where a plot ln (reactants) against time produces a straight line if the
reaction is first order (Fig. 49.2, p. 451).
(a) 5 a = +1
Linear functions and straight lines
4 a=0
One of the most straightforward and widely used relationships between two
y variable
3 a = -1
variables x and y is that represented by a straight-line graph, where the cor-
2
responding mathematical function is known as the equation of a straight line,
1 where:
0
-1 1 2 3 4 y = a + bx[51.3]
-1
x variable
(b) 8 b = +2 In this relationship, a represents the intercept of the line on the y-(vertical)
axis, i.e. where x = 0, and therefore bx = 0, while b is equivalent to the slope
6
(gradient) of the line, i.e. the change in y for a change in x of 1. The constants
y variable
4 b = +1 a and b are sometimes given alternative symbols, but the mathematics remains
unchanged, e.g. in the equivalent expression for the slope of a straight line,
2 b = +0.5
y = mx + c. Figure 51.1 shows what happens when these two constants are
0 changed, in terms of the resultant straight lines.
0 1 2 3 4
x variable The two main applications of the straight-line relationship are:
Fig. 51.1 Straight-line relationships 1. Function fitting. Here, you determine the mathematical form of the func-
(y = a + bx), showing the effects of tion, i.e. you estimate the constants a and b from a data set for x and y,
(a) changing the intercept at constant either by drawing a straight line by eye and then working out the slope
slope, and (b) changing the slope at con- and y intercept, or by using linear regression (p. 499) to obtain the most
stant intercept.

(a) 2.5 probable values for both constants. When putting a straight line of best fit
2 by eye on a hand-drawn graph, note the following:
1.5 Always use a transparent ruler, so you can see data points on either
y variable
●●
1 -1.6 side of the line.

0.5 ●● For a data series where the points do not fit a perfect straight line,
+4.0
0 try to have an equal number of points on either side of the line, as
0 1 2 3 4 5
x variable in Fig. 51.2(a), and try to minimise the average distance of these points
(b) 20 from the line.
tangent touches at
15
exactly x = 1.8 ●● Once you have drawn the line of best fit use this line, rather than
your data values, in all subsequent procedures (e.g. in a calibration
y variable
10 +16 curve, Chapter 48).

5
+4.0
●● Tangents drawn to a curve give the slope (gradient) at a particular
0
point, e.g. in a titration curve. These are best drawn by bringing your
0 2 4 6 8 ruler up to the curve at the exact point where you wish to estimate the
x variable slope and then trying to make the two angles immediately on either side
Fig. 51.2 Drawing straight lines. (a) Sim-
of this point approximately the same, by eye (Fig. 51.2(b)).
ple linear relationship, giving a straight ●● Once you have drawn the straight line or tangent, choose two points
line with an intercept of 2.3 and a slope reasonably far apart at either end of your line and then draw construction
of - 1.6 , 4.0 = 0.4. (b) Tangent drawn lines to represent the change in y and the change in x between these two
to a curve at x = 1.8, giving a slope of
points: make sure that your construction lines are perpendicular to each
16 , 4 = 4.
other. Determine the slope as the change in y divided by the change in
x (Fig. 51.2(b)).
2. Prediction. Where a and b are known, or have been estimated, you can
use eqn [51.3] to predict any value of y for a specified value of x, e.g. in
Example Using eqn [51.4], the a calibration curve. You will need to rearrange eqn [51.3] in cases where
predicted value for x for a lin- a prediction of x is required for a particular value of y (e.g. in calibration
ear function where a = 1.5 and curves, p. 446), as follows:
b = 2.5, where y = - 8.5 is:
x = ( - 8.5 - 1.5) , 2.5 = -4. x = (y - a) , b[51.4]
Using eqn [51.4] the predicted
x intercept for a linear function This equation can also be used to determine the intercept on the
where a = 0.8 and b = 3.2 is:
x-(horizontal) axis, i.e. where y = 0.
x = (0 - 0.8) , 3.2 = - 0.25.
Hints for some typical problems
Example A lab schedule states that

Calculations involving proportions or ratios
5 g of a compound with a relative The ‘unitary method’ is a useful way of approaching calculations involving
molecular mass of 220 are dissolved proportions or ratios, such as those required when making up solutions from
in 400 mL of solvent. For writing up stocks (see also Chapter 8) or as a subsidiary part of longer calculations.
your Materials and Methods, you wish
to express this as mol L-1. 1. If given a value for a multiple, work out the corresponding value for
a single item or unit.
1. If there are 5 g in 400 mL, then
there are 5 , 400 g in 1 mL. 2. Use this ‘unitary value’ to calculate the required new value.
2. Hence, 1000 mL will contain Calculations involving series

5 , 400 * 1000 g = 12.5 g. Series (used in, for example, dilutions, see also p. 75) can be of three main
3. 12.5 g = 12.5 , 220 mol = forms:
0.0568 mol, so [solution] =
1. Arithmetic, where the difference between two successive numbers in the
56.8 mmol L-1 ( = 56.8 mol m -3).
series is a constant, e.g. 2, 4, 6, 8, 10, . . .

2. Geometric, where the ratio between two successive numbers in the series
is a constant, e.g. 1, 10, 100, 1000, 10 000, . . .
Examples For a geometric dilu-
tion series involving tenfold dilution 3. Harmonic, where the values are reciprocals of successive whole numbers,
steps, calculation of concentrations is e.g. 1, 12, 13, 14, c
straightforward, e.g. two serial dec-
imal dilutions ( = 100@fold dilution)
Note that the logs of the numbers in a geometric series will form an arith-
of a solution of NaCl of 250 mmol L-1 metic series (e.g. 0, 1, 2, 3, 4, . . . in the above case). Thus, if a quantity y varies
will produce a dilute solution of with a quantity x such that the rate of change in y is proportional to the value
250 , 100 = 2.5 mmol L-1. Similarly, of y (i.e. it varies in an exponential manner), a semi-log plot of such data will
for an arithmetic dilution series, divide form a straight line. This form of relationship is relevant for radioactive decay
by the overall dilution to give the final (p. 311).
concentration, e.g. a 16-fold dilution
of a solution of NaCl of 200 mg mL-1 Statistical calculations
will produce a dilute solution of The need for long, complex calculations in statistics has largely been removed
200 , 16 = 12.5 mg mL-1. because of the widespread use of spreadsheets with statistical functions
(Chapter 45) and specialised programs such as SPSS and Minitab. It is, how-
ever, important to understand the principles behind what you are trying to do
(see Chapters 53 and 54) and interpret the program’s output correctly, either
using the ‘help’ function or a reference manual.
Text references
Adams, D.S. (2003) Lab Math: A Handbook of Measure- Eason, G., Coles, C.W. and Gettinby, G. (1992) Mathemat-
ment, Calculations, and Other Quantitative Skills for ics and Statistics for the Bio-Sciences. Ellis Horwood,
Use at the Bench. Cold Spring Harbor Lab Publishing, Chichester.
Cold Spring Harbor.

Anon. S.O.S. Mathematics. Kirkup, L. (1994) Experimental Methods: An Introduction
Available: http://www.sosmath.com to the Analysis and Presentation of Data. John Wiley &
Last accessed 01/09/15. Sons Ltd, New York.
[A basic web-based guide with very wide coverage.] Lawler, G. (2011) Understanding Maths. Basic Mathemat-
Billo, E.J. (2007) Excel for Scientists and Engineers. John ics Explained. Studymates, 4th edn. Aber Publishing,
Wiley & Sons Ltd, Chichester. Abergele.
[Covers basic principles, units, logarithms, ratios and Miller, J.N. and Miller, J.C. (2010) Statistics and Chemom-
p roportions, concentrations, equations, rates and
etrics for Analytical Chemistry, 6th edn. Prentice Hall,
graphs.] Harlow.
Currell, G. and Dowman, T. (2009) Essential Mathematics Monk, P. and Munro, L.J. (2010) Maths for Chemists: A
and Statistics for Science, 2nd edn. John Wiley & Sons Chemist’s Toolkit of Calculations, Oxford University
Ltd, Chichester. Press, Oxford.
Forster, P.C. (1999) Easy Mathematics for Biologists. Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
Harwood, Amsterdam. Oxford University Press, Oxford.

Study exercises
51.1 Rearrange a simple formula. The Beer–Lambert (e) 99.897 to two decimal places
relationship, eqn [29.3] is written in the form (f) 99.997 to two decimal places
A = e/[C]. Rearrange, in the form: (g) 6255 to the nearest 10
(h) 134 903 to the nearest ten thousand
(a) [C] =
(b) e = State the following:
51.2 Rearrange the following formulae: (i) the number of significant figures in 3400
(j) the number of significant figures in 3400.3
(a) If y = ax + b, find b
(k) the number of significant figures in 0.001 67
(b) If y = ax + b, find x
(l) the number of significant figures in 1.001 67
(c) If x = y3, find y
(m) the number of decimal places in 34.46
(d) If x = 3y, find y
(n) the number of decimal places in 0.001 67
(e) lf x = (1 - y)(zp + 3), find z
(f) If x = (y - z)1/n/pq, find n 51.4 Carry out calculations involving percentages.
Answer the following questions, giving your
51.3 Work with decimal places or significant figures.
answers to two decimal places:
Give the following numbers to the accuracy
indicated: (a) What is 6/35ths expressed as a percentage?
(b) What is 0.0755 expressed as a percentage?
(a) 214.51 to three significant figures
(c) What is 4.35% of 322?
(b) 107 029 to three significant figures
(d) A rat’s weight increased from 55.23 to 75.02 g.
(c) 0.0450 to one significant figure
What is the % increase in its weight?
(d) 99.817 to two decimal places

52 Manipulating and transforming raw data
The process of discovering the meaning within your results can be fascinating.
There are two main elements to this process.
1. Exploratory data manipulation – this is used to investigate the nature of
your results and suggest possible patterns and relationships within the data
set. The aim is to generate hypotheses for further investigation. Explora-
tory techniques allow you to visualise the form of your data. They are ideal
for examining results from pilot ‘studies’, but should be used throughout
your investigations.
2. Confirmatory analysis – this is used to test the hypotheses generated dur-
ing the exploratory phase. The techniques required are generally statistical
in nature and are dealt with in Chapter 54.
Key Point Spreadsheets (Chapter 45) are invaluable tools for

data manipulation and transformation: complex mathematical
procedures can be carried out rapidly and the results visual-
ised almost immediately using the inbuilt graphing functions.
Spreadsheets also facilitate the statistical analysis of data (see
Chapters 53 and 54).
Summarising your results – original,

unsummarised data belong only in Organising numbers
your primary record, either in labo- To organise, manipulate and summarise data, you should:
ratory books or as computer records.
You should produce summary tables ●● simplify the numbers, e.g. by rounding or taking means. This avoids the
to organise and condense original data. detail becoming overwhelming;
●● rearrange your data in as many ways as possible for comparison;
●● display in graphical form; this provides an immediate visual summary that
Colour Tally Total is relatively easy to interpret;
●● look for an overall pattern in the data – avoid getting lost in the details at
this stage;
Green III 3
Blue IIII III 8 ●● look for any striking exceptions to that pattern (outliers) – they often
point to special cases of particular interest or to errors in the data produced
Red IIII 4 through mistakes during the acquisition, recording or copying of data;
White IIII IIII II 12 ●● move from graphical interpretations to well-chosen numerical summa-
Black I 1 ries and/or verbal descriptions, including, where applicable, an explanatory
Maroon III 3 hypothesis.
Yellow II 2 After collecting data, the first step is often to count how frequently each
33 value occurs and to produce a frequency table. The frequency is simply the
number of times a value occurs in the data set, and is, therefore, a count. The
raw data could be acquired using a tally chart system to provide a simple fre-
Fig. 52.1 An example of a tally chart. quency table. To construct a tally chart (e.g. Fig. 52.1):
●● enter only one tally at a time;
Producing a histogram – a neatly con- ●● if working from a data list, cross out each item on the list as you enter it
structed tally chart will double as a on to the tally chart, to prevent double entries;
rough histogram.
●● check that all values are crossed out at the end and that the totals agree.

Manipulating and transforming raw data
Table 52.1 An example of a frequency Convert the data to a formal table when complete (e.g. Table 52.1).
table. Because proportions are easier to compare than class totals, the table may con-
tain a column to show the relative frequency of each class. Relative frequency
Relative can be expressed in decimal form (as a proportion of 1) or as a percentage (as
Size class Frequency frequency (%)
a proportion of 100).
0–4.9 7 2.6
5–9.9 23 8.6 Graphing data
10–14.9 56 20.9
Graphs are an effective way to investigate trends in data and can reveal features
15–19.9 98 36.7 that are difficult to detect from a table, e.g. skewness of a frequency distribu-
20–24.9 50 18.7 tion. The construction and use of graphs is described in detail in Chapter 49.
25–29.9 30 11.2
When investigating the nature of your data, the main points are as follows:
30–34.9 3 1.1 ●● make the values stand out clearly: attention should focus on the actual
Total 267 99.8* data, not the labels, scale markings, etc. (contrast with the requirements for
constructing a graph for data presentation; see p. 451);
* ≠ 100 because of rounding error.
●● avoid clutter in the graph: leave out grid lines and try to use the simplest
graph possible for your purpose.
Key Point Use a computer spreadsheet with graphics options

whenever possible: the speed and flexibility of these powerful
tools should allow you to explore every aspect of your data rap-
idly and with relatively little effort (see Chapters 45 and 51).
stem leaves
7 23 Displaying distributions
7 55
7 6 A visual display of a distribution of values is often useful for variables meas-
7 9 ured on an interval or ratio scale (p. 452). The distribution of a variable can
8 000 be displayed by a frequency table for each value or, if the possible values are
8 233 numerous, groups (classes) of values of the variable. Graphically, there are two
8 45555 main ways of viewing such data:
8 77
8 888899 1. Histograms (see pp. 452), generally used for large samples.
9 0000111111 2. Stem and leaf plots (e.g. Fig. 52.2), often used for samples of less than
9 2333333
100: these retain the actual values and are faster to draw by hand. The main
9 44555555555
9 66677777 drawback is the limitation imposed by the choice of stem values since these
9 88888999 class boundaries may obscure some features of the distribution.
10 00 These displays allow you to look at the overall shape of a distribution and
to observe any significant deviations from the idealised theoretical ones. Where
Fig. 52.2 A simple ‘stem and leaf’ plot of necessary, you can use data transformations to investigate any departures from
a data set. The ‘stem’ shows the common standard distribution patterns such as the normal distribution or the Poisson
component of each number, while the distribution (p. 493).
‘leaves’ show the individual components,
e.g. the top line in this example represents
the numbers 72 and 73. Transforming data
Transformations are mathematical functions applied to data values. They are
particularly valuable where your results are related to areas and volumes (e.g.
field area, sample size).
The most common use of transformations is to prepare data sets so that
specific statistical tests may be applied. For instance, if you find that your data
distribution is unimodal but not symmetrical (p. 486), it is often useful to apply
a transformation that will redistribute the data values to form a symmetrical

distribution. The object of this exercise is often to find the function that most
Using transformations – note that if
closely changes the data into a standard normal distribution, allowing you to
you wish to conserve the order of your
apply a wide range of parametric hypothesis-testing statistics (see Chapter 54).
data, you will need to take negative
A frequently used transformation is to take logarithms of one or more sets of
values when using a reciprocal func-
values: if the data then approximate to a normal distribution, the relationship
tion (i.e. - 1/nn). This is essential when
is termed ‘log-normal’.
using a box plot to compare graphically
Some general points about transformations are:
the effects of transformations on the
five-number summary of a data set. ●● they should be made on the raw data, not on derived data values: this is
simpler, mathematically valid and more easily interpreted;
●● the transformed data can be analysed like any other numbers;
Transformation Effect
●● transformed data can be examined for outliers, which may be more
1/x 2
increasingly strong important if they remain after transformation.
correction for positive
1/x
skew in data set Figure 52.3 presents a ladder of transformations that will help you decide
which transformations to try (see also Table 54.1). Note that percentage and
proportion data are usually arc-sine transformed, which is a more complex
log x procedure.
Figure 52.4 illustrates the following ‘quick-and-easy’ way to choose a
x transformation:
1. Calculate the ‘five-number summary’ for the untransformed data (p. 485).
x no shape
change 2. Present the summary graphically as a ‘box and whisker’ plot (p. 485).
3. Decide whether you need to correct for positive or negative skew
x2
(p. 486).
x3
4. Apply one of the ‘mild’ transformations in Fig. 52.3 on the five-number
summary values only.
antilog x increasingly strong 5. Draw a new box and whisker plot and see whether the skewness has been
correction for negative
skew in data set
corrected.
6. If the skewness has been undercorrected, try again with a stronger
Fig. 52.3 Ladder of transformations (after transformation. If it has been overcorrected, try a milder one.
J.W. Tukey).
7. When the distribution appears to be acceptable, transform the full
data set and recalculate the summary statistics. If necessary, use a
Original data set box and whisker plots Transformed data set
2
Frequency
Frequency
original 1st 2nd

Dependent variable Dependent variable
data set transformation transformation
(transformed)
–positively which leads to
skewed overcompensates near-symmetrical
–reject distribution
–accept
Fig. 52.4 Illustration of the processes of transforming a data set.

statistical test to confirm that the transformed data are normally dis-
tributed (p. 492).
8. If no simple transformation works well, you may need to use non-
parametric statistics when comparing data sets.

Currell, G. and Dowman, T. (2009) Essential Mathematics Schmuller, J. (2013) Statistical Analysis with Excel for
and Statistics for Science, 2nd edn. John Wiley & Sons Dummies John Wiley & Sons Ltd, Hoboken.
Ltd, Chichester. Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
Dytham, C. (2010) Choosing and Using Statistics: A Biol- Oxford University Press, Oxford.
ogists’ Guide, 2nd edn. Blackwell, Oxford. Taylor, J.K. and Cihon, C. (2004) Statistical Techniques
Miller, J.N. and Miller, J.C. (2010) Statistics and Chemomet- for Data Analysis, 2nd edn. CRC Press, Boca Raton.
rics for Analytical Chemistry 6th edn. Prentice Hall, Harlow.
Monk, P. and Munro, L.J. (2010) Maths for Chemists: A
Chemist’s Toolkit of Calculations. Oxford University
Press, Oxford.
Study Exercises
The following study exercises can be most easily car- 52.2 Use a stem and leaf plot. Work out the mean and
ried out using a spreadsheet such as Microsoft Excel. standard deviation of the data contained in the
following stem and leaf plot:
52.1 Compare frequency distributions for samples
with different sample sizes. Compute relative 6 1334
frequencies for each class in the table below. 6 56788
Graph the data as a frequency polygon. 7 013344
Frequency distribution data. 7 556667899
8 11224
Hyphal Treat- Treat- Hyphal Treat- Treat-
8 569
length ment ment length ment ment
(Mm) B A (Mm) B A 9 03
9 6
0 0 0 13 6 30
1 1 0 14 3 26 Stem and leaf plot.
2 2 0 15 1 22
52.3 Use transformations to correct skew in a data set.
3 4 1 16 1 18
Find a transformation that will make the data of
4 8 1 17 0 14
treatment B in Study exercise 52.1 approximately
5 13 2 18 0 12
symmetrical about the mean. Demonstrate
6 22 5 19 0 8
graphically that you have accomplished this.
7 36 9 20 0 5
8 48 14 21 0 4
9 46 21 22 0 2
10 32 26 23 0 2
11 18 31 24 0 1
12 11 33 25 0 1

53 Descriptive statistics
The purpose of most practical work is to observe and measure a particular char-
Use of symbols – Y is used in Chapters
acteristic of a molecule or chemical system. However, it would be extremely
53 and 54 to signify the dependent var-
rare if the same value was obtained every time the characteristic was meas-
iable in statistical calculations. Note,
ured, or with every experimental subject. More commonly, such measurements
however, that some calculators refer
will show variability, caused by measurement error, sampling variation and/or
to, e.g., x, Σx2, etc. for their statistical
chemical variability (p. 38). Such variability can be displayed as a frequency
functions.
distribution (e.g. Fig. 50.2), where the y-axis shows the number of times (fre-
quency, f) each particular value of the measured (dependent) variable (Y) has
been obtained. Descriptive (or summary) statistics quantify aspects of the fre-
quency distribution of a sample. You can use them to condense a large data set,
for presentation in figures or tables. An additional application of descriptive
statistics is to provide estimates of the true values of the underlying frequency
distribution of the population being sampled, allowing the significance and
precision of the observations to be assessed (pp. 486, 492).
Summarising your results – original Key Point The appropriate descriptive statistics to choose
data belong only in your primary will depend on both the type of data, i.e. whether quantitative,
record, either in laboratory books or ranked or qualitative (p. 36), and the nature of the underlying
as computer records. You should pro- frequency distribution.
duce summary tables to condense and
describe original data.
In many instances, the normal (Gaussian) distribution (Chapter 54) best
describes the observed pattern for a quantitative variable, giving a symmetrical,
bell-shaped frequency distribution (p. 494); for example, measurements of a
quantitative continuous variable across a number of sites (e.g. PAH concentra-
tion across a contaminated land site), or replicate measurements of a particular
characteristic (e.g. repeated measurements of PAH concentration in an specific
sampling location within the site). If you have no clear theoretical grounds for
assuming what the underlying frequency distribution is like, graph one or more
sample frequency distributions, ideally with a sample size7 100.
Three important features of a frequency distribution that can be summa-
rised by descriptive statistics are:
1. the sample’s location, i.e. its position along a given dimension represent-
ing the dependent (measured) variable (Fig. 53.1);
A B
2. the dispersion of the data, i.e. how spread out the values are (Fig. 53.2);
3. the shape of the distribution, i.e. whether symmetrical, skewed, U-shaped
(Fig. 53.3).
Frequency
Measuring location
Here, the objective is to pinpoint the ‘centre’ of the frequency distribution, i.e.
the value about which most of the data are grouped. The chief measures of
location are the mean, median and mode. Fig. 53.4 shows how to choose among
Dependent (measured) variable
these for a given data set.
Fig. 53.1 Two distributions with different
Mean
locations but the same dispersion. The data
set labelled B could have been obtained by The mean (denoted Y and also referred to as the arithmetic mean) is the average
adding a constant to each datum in the value of the data. It is obtained from the sum of all the data values divided by
data set labelled A. the number of observations (in symbolic terms, ΣY/n). The mean is a good

Descriptive statistics
median
mean
mode
A
mode
mode
median
median
mean
mean
B
Frequency
Frequency
(a) (b) (c)
symmetrical positively negatively
skewed skewed
Dependent (measured) variable Dependent (measured) variable
Fig. 53.2 Two distributions with different Fig. 53.3 Symmetrical and skewed frequency distributions, showing rela-
dispersions but the same location. The data tive positions of mean, median and mode.
set labelled A covers a relatively narrow
range of values of the dependent (meas-
ured) variable while that labelled B covers WHAT TYPE OF VARIABLE
a wider range. AM I SAMPLING?
quantitative ranked qualitative

(continuous or
discontinuous)
mode mode
WHAT IS THE SHAPE mean (with
OF THE DISTRIBUTION? caution)
one main peak two or more

pronounced peaks
symmetrical asymmetrical outliers

present
Example Box 53.1 shows a set of
data and the calculated values of the
mean median modes
measures of location, dispersion and median mean (if Poisson) median median (with
shape for which methods of calcu- mode mode mode mean caution)
lation are outlined here. Check your
understanding by calculating the Fig. 53.4 Choosing a statistic for characterising a distribution’s location.
statistics yourself and confirming that Statistics written in bold are the preferred option(s).
you arrive at the same answers.
measure of the centre of symmetrical frequency distributions. It uses all of the

numerical values of the sample and therefore incorporates all of the information
Definitions content of the data. However, the value of a mean is greatly affected by the
presence of extreme values (outliers). The arithmetic mean is a widely used
An outlier – any datum that has a value statistic in the biosciences, but there are situations when you should be careful
much smaller or bigger than most of the about using it (see Box 53.2 for examples).
other data values.
Rank – the position of a data value Median
when all the data are placed in order The median is the mid-point of the observations when ranked in increasing order.
of ascending magnitude. If ties occur, For odd-sized samples, the median is the middle observation; for even-sized
an average rank of the tied variates is samples it is the mean of the middle pair of observations. Where data are grouped
used. Thus, the rank of the datum 6 in into classes, the median can only be estimated. This is most simply done from
the sequence 1, 3, 5, 6, 8, 8, 10 is 4; the
a graph of the cumulative frequency distribution, but can also be worked out by
rank of each datum with value 8 is 5.5.
assuming the data to be evenly spread within the class. The median may represent

Box 53.1 Descriptive statistics for a sample of data – an example
Value (Y) Frequency (f ) Cumulative fY fY 2

frequency
1 0 0 0 0
2 1 1 2 4
3 2 3 6 18
4 3 6 12 48
5 8 14 40 200
6 5 19 30 180
7 2 21 14 98
8 0 21 0 0
Totals 21 = a f ( = n) 104 = a fY 548 = a fY 2
In this example, for simplicity and ease of calculation, underlying frequency distribution of such data sets,
integer values of Y are used. In many practical exer- you would need to group individual data into broader
cises, where continuous variables are measured to classes (e.g. all values between 1.0 and 1.9, all values
several significant figures and where the number of between 2.0 and 2.9, etc.) and then draw a histogram
data values is small, giving frequencies of 1 for most (p. 458). Calculation of certain statistics for data sets
of the values of Y, it may be simpler to omit the column that have been grouped in this way (e.g. median, quar-
dealing with frequency and list all the individual values tiles, extremes) can be tricky, and a statistical text
of Y and Y 2 in the appropriate columns. To gauge the should be consulted.
Statistic Value* How calculated
a fY/n, i.e. 104/21

Mean 4.95
Median 5 Value of the (n + 1)/2 variate, i.e. the value ranked (21 + 1)/2 = 11th
(obtained from the cumulative frequency column)
Mode 5 The most common value (Y value with highest frequency)
Upper quartile 6 The upper quartile is between the 16th and 17th values, i.e. the value
exceeded by 25% of the data values
Lower quartile 4 The lower quartile is between the 5th and 6th values, i.e. the value
exceeded by 75% of the data values
Semi-interquartile range 1.0 Half the difference between the upper and lower quartiles, i.e. (6 - 4)/2
Upper extreme 7 Highest Y value in data set
Lower extreme 2 Lowest Y value in data set
Range 5 Difference between upper and lower extremes
Variance (s2) 1.65 Σf Y 2 - 1 Σf Y 2 2/n
s2 =
n - 1
548 - (104)2/21
=
20
Standard deviation (s) 1.28 Us 2
Standard error (SE) 0.280 s/Un
95% confidence limits 4.36–5.54 Y | t0.05[20] * SE, (where t0.05[20] = 2.09, Table 54.2)
Coefficient of variation (cov) 25.9% 100s/Y
*Rounded to three significant figures (see p. 470), except when it is an exact number.

Box 53.2 Three examples where simple arithmetic means are inappropriate
1. If means of samples are themselves meaned, an error can arise if the

Mean n
samples are of different size. For example, the arithmetic mean of
6 4 the means in the table shown left is 7, but this does not take account
of the different ‘reliabilities’ of each mean owing to their sample
7 7
sizes. The correct weighted mean is obtained by multiplying each
8 1 mean by its sample size (n) (a ‘weight’) and dividing the sum of these
values by the total number of observations, i.e. in the case shown,
(24 + 49 + 8)/12 = 6.75.
2. When making a mean of ratios (e.g. percentages) for several groups
of different sizes, the ratio for the combined total of all the groups is
not the mean of the proportions for the individual groups. For exam-
ple, if 20 tablets from a batch of 50 are yellow, this implies 40% are
yellow. If 60 tablets from a batch of 120 are yellow, this implies 50%
are yellow. The mean percentage of yellow tablets (50 + 40)/2 = 45%
is not the percentage of yellow tablets in the two groups combined,
because there are 20 + 60 = 80 yellow tablets in a total of 170
tablets = 47.1% approx.
pH value [H+ ](mol L −1) 3. If the measurement scale is not linear, arithmetic means may give
a false value. For example, if three media had pH values 6, 7 and 8,
6 1 * 10 -6 the appropriate mean pH is not 7 because the pH scale is logarith-
7 1 * 10 -7 mic. The definition of pH is -log10[H +], where [H +] is expressed in
mol l-1 (‘molar’); therefore, to obtain the true mean, convert data into
8 1 * 10 -8
[H +] values (i.e. put them on a linear scale) by calculating 10(-pH value)
mean 3.7 * 10 -7 as shown. Now calculate the mean of these values and convert the
- log10 mean 6.43 answer back into pH units. Thus, the appropriate answer is pH 6.43
rather than 7. Note that a similar procedure is necessary when cal-
culating statistics of dispersion in such cases, so you will find these
almost certainly asymmetric about the mean.
Mean values of log-transformed data are often termed geometric means –
they are sometimes used in biochemistry, where log-transformed val-
ues for antibody concentrations in human blood serum are averaged
and plotted, rather than using the raw data values. The use of geometric
means in such circumstances serves to reduce the effects of outliers on
the mean.
the location of the main body of data better than the mean when the distribution
is asymmetric or when there are outliers in the sample.
Mode
The mode is the most common value in the sample. The mode is easily found
from a tabulated frequency distribution as the most frequent value. If data have
been grouped into classes then the term modal class is used for the class con-
taining most values. The mode provides a rapidly and easily found estimate of
Describing the location of qualitative
data – the mode is the only statistic
sample location and is unaffected by outliers. However, the mode is affected by
that is suitable for this task. For exam-
chance variation in the shape of a sample’s distribution and it may lie distant
ple, the modal (most frequent) titre
from the obvious centre of the distribution.
value was 21.2 mL.
The mean, median and mode have the same units as the variable under dis-
cussion. However, whether these statistics of location have the same or similar
values for a given frequency distribution depends on the symmetry and shape
of the distribution. If it is near-symmetrical with a single peak, all three will be

very similar; if it is skewed or has more than one peak, their values will differ
to a greater degree (see Fig. 53.3).
Measuring dispersion
Here, the objective is to quantify the spread of the data about the centre of the
distribution. Figure 53.5 indicates how to decide which measure of dispersion
to use.
Range
The range is the difference between the largest and smallest data values in the
Example In a sample of data with sample (the extremes) and has the same units as the measured variable. The
values 3, 7, 15, 8, 5, 10 and 4, the range
range is easy to determine, but is greatly affected by outliers. Its value may
is 12 (i.e. the difference between the
highest value, 15, and the lowest
also depend on sample size: in general, the larger this is, the greater will be the
value, 3). range. These features make the range a poor measure of dispersion for many
practical purposes.
Semi-interquartile range
The semi-interquartile range is an appropriate measure of dispersion when a
median is the appropriate statistic to describe location. For this, you need to
determine the first and third quartiles, i.e. the medians for those data values
ranked below and above the median of the whole data set (see Fig. 53.6). To
calculate a semi-interquartile range for a data set:
1. rank the observations in ascending order;
2. find the values of the first and third quartiles;
3. subtract the value of the first quartile from the value of the third;
4. halve this number.
WHAT TYPE OF VARIABLE

AM I SAMPLING?
quantitative ranked
(continuous or or
discontinuous) qualitative
WHAT IS THE SHAPE

OF THE DISTRIBUTION? no meaningful
statistics of
dispersion can
be applied
symmetrical asymmetrical outliers

present
standard deviation semi-interquartile range

semi-interquartile five-number summary semi-interquartile range
range variance (if Poisson) five-number summary
Fig. 53.5 Choosing a statistic for characterising a distribution’s disper-

sion. Statistics written in bold are the preferred option(s). Note that you
should match statistics describing dispersion with those you have used
to describe location, i.e. standard deviation with mean, semi-interquartile
range with median.

lower upper For data grouped in classes, the semi-interquartile range can only be
extreme extreme
estimated. Another disadvantage is that it takes no account of the shape of
range
the distribution at its edges. This objection can be countered by using the
lowest full set of data, ranked in order highest
so-called ‘five number summary’ of a data set, which consists of the three
quartiles and the two extreme values; this can be presented on graphs as
lower half upper half
a box and whisker plot (see Fig. 53.7) and is particularly useful for sum-
marising skewed frequency distributions. The corresponding ‘six number
summary’ includes the sample’s size.
lower median upper

quartile (middle quartile Variance and standard deviation
quartile)
For symmetrical frequency distributions, an ideal measure of dispersion
interquartile
range would take into account each value’s deviation from the mean and provide
the sample variance, which is the sum of squares ( a (Y - Y )2) divided

a measure of the average deviation from the mean. Two such statistics are
semi-interquartile
range = 1/2 interquartile range
by n - 1 (where n is the sample size), and the sample standard deviation,
Fig. 53.6 Illustration of median, quartiles,
which is the positive square root of the sample variance.
range and semi-interquartile range.
The variance (s2) has units which are the square of the original units,
while the standard deviation ( s, or SD) is expressed in the original units, one
upper extreme reason s is often preferred as a measure of dispersion. Calculating s or s2 long-
hand is a tedious job and is best done with the help of a calculator or computer.
upper quartile If you do not have a calculator that calculates s for you, an alternative formula
that simplifies calculations is:
median ΣY 2 - (ΣY )2/n

s = + [53.1]
C n - 1
To calculate s using a calculator:
1. Obtain ΣY, square it, divide by n and store in memory;

lower quartile 2. Square Y values, obtain ΣY 2, subtract the memory value from this;
3. Divide this answer by n - 1.
lower extreme 4. Take the positive square root of this value.
Fig. 53.7 A box and whisker plot, showing Take care to retain significant figures, or errors in the final value of s will
the ‘five number summary’ of a sample as result. If continuous data have been grouped into classes, the class mid-values
it might be used on a graph. or their squares must be multiplied by the appropriate frequencies before
summation (see example in Box 53.1). When data values are large, long-
hand calculations can be simplified by coding the data, e.g. by subtracting
a constant from each datum, and decoding when the simplified calculations
are complete.
Using a calculator for statistics – make
sure you understand how to enter indi-
vidual data values and which keys will
Coefficient of variation
give the sample mean (usually shown as The coefficient of variation (CoV) is a dimensionless measure of variability
X or x) and sample standard deviation relative to location which expresses the sample standard deviation, usually as
(often shown as sn - 1). In general, you a percentage of the sample mean, i.e.
should not use the population standard
deviation (usually shown as sn). CoV = 100s/Y(%)[53.2]
This statistic is useful when comparing the relative dispersion of data sets
with widely differing means, or where different units have been used for the
same or similar quantities.

A useful application of the CoV is to compare different analytical methods

or procedures, so that you can decide which involves the least proportional
Example Consider two methods of
error – create a standard stock solution, then base your comparison on the
analysis for a Pb in soil. For a given
standard, Method A gives a mean
results from several subsamples analysed by each method. You may find it
result of = 50 ‘response units’ with useful to use the CoV to compare the precision of your own results with those
s = 8, while Method B gives a mean of a manufacturer, e.g. for a pipettor (p. 65). The smaller the CoV, the more
result of = 160 ‘response units’ with precise (repeatable) is the apparatus or technique. (Note: this does not mean
s = 18. Which analysis gives the more that it is necessarily more accurate, see p. 38.)
reproducible results? The answer can
be found by calculating the CoV val- Measuring the precision of the sample mean as an
ues, which are 16 and 11.25%, respec-
estimate of the true value using the standard error
tively. Hence, Method B is the more
precise (K reproducible), even though Most practical exercises are based on a limited number of individual data values
the absolute value of s is larger. (a sample) which are used to make inferences about the population from which
they were drawn. For example, the Pb content might be measured in blood sam-
ples from 100 adults and used as an estimate of the adult blood Pb content, with
m the sample mean (Y) and sample standard deviation (s) providing estimates of
the true values of the underlying population mean (m) and the population stand-
ard deviation (s). The reliability of the sample mean as an estimate of the true
(population) mean can be assessed by calculating a statistic termed the standard
SE SE error of the sample mean (often abbreviated to standard error or SE), from:
SE = s/2n[53.3]
Strictly, the standard error is an estimate of the dispersion of repeated

sample means around the true (population) value: if several samples were taken,
Fig. 53.8 Frequency distribution of sample each with the same number of data values (n), then their means would cluster
means around the population mean (m). around the population mean (m) with a standard deviation equal to SE, as
Note that SE is equivalent to the standard shown in Fig. 53.8. Therefore, the smaller the SE, the more reliable the sam-
deviation of the sample means, for sample
ple mean is likely to be as an estimate of the true value, since the underlying
size = n.
frequency distribution would be more tightly clustered around m. At a practical
level, Eqn [53.3] shows that SE is directly affected by the dispersion of indi-
vidual data values within the sample, as represented by the sample standard
deviation (s). Perhaps more importantly, SE is inversely related to the square
root of the number of data values (n). Therefore, if you wanted to increase the
precision of a sample mean by a factor of 2 (i.e. to reduce SE by half), you
would have to increase n by a factor of 22 (i.e. fourfold).
Summary descriptive statistics for the sample mean are often quoted as
Example Summary statistics for the
sample mean and standard error for Y { SE(n), with the SE being given to one significant figure more than the
the data shown in Box 53.1 would be mean. You can use such information to carry out a t-test between two sample
quoted as 4.95 { 0.280 (n = 21). means (Box 54.1); the SE is also useful because it allows calculation of confi-
dence limits for the sample mean (p. 499).
Describing the ‘shape’ of frequency distributions

Frequency distributions may differ in the following characteristics:
●● number of peaks;
●● skewness or asymmetry;
●● kurtosis or pointedness.
The shape of a frequency distribution of a small sample is affected by
chance variation and may not be a fair reflection of the underlying population

(a) unimodal (b) bimodal (c) polymodal
Frequency
Fig. 53.9 Frequency distributions with different numbers of peaks. A uni-

modal distribution (a) may be symmetrical or asymmetrical. The dotted
lines in (b) indicate how a bimodal distribution could arise from a combi-
nation of two underlying unimodal distributions. Note here how the term
‘bimodal’ is applied to any distribution with two major peaks – their fre-
quencies do not have to be exactly the same.
frequency distribution: check this by comparing repeated samples from the

same population or by increasing the sample size. If the original shape were
caused by random events, it should not appear consistently in repeated samples
and should become less obvious as sample size increases.
Genuinely bimodal or polymodal distributions may result from the
combination of two or more unimodal distributions, indicating that more
Calculating the extent of skew and kur-
than one underlying population is being sampled (Fig. 53.9). An example
tosis of a data set – use the SKEW and
of a bimodal distribution is the height of adult humans (females and males
KURT functions in Microsoft Excel.
combined).
A distribution is skewed if it is not symmetrical, a symptom being that
the mean, median and mode are not equal (Fig. 53.3). Positive skewness is
where the longer ‘tail’ of the distribution occurs for higher values of the meas-
ured variable; negative skewness where the longer tail occurs for lower values.
Some chemical examples of characteristics distributed in a skewed fashion are
pollutants levels in soil samples, insulin levels in human plasma and pesticide
(b) leptokurtic levels in agricultural products.
Frequency
(a) platykurtic
Kurtosis is the name given to the ‘pointedness’ of a frequency distribu-
tion. A platykurtic frequency distribution is one with a flattened peak, while
a leptokurtic frequency distribution is one with a pointed peak (Fig. 53.10).
While descriptive terms can be used, based on visual observation of the
shape and direction of skew, the degree of skewness and kurtosis can be
quantified and statistical tests exist to test the ‘significance’ of observed
Fig. 53.10 Examples of the two types of values, but the calculations required are complex and best done with the aid
kurtosis. of a computer.
Using computers to calculate descriptive statistics

There are many specialist statistical packages (e.g. SPSS) that can be used to
simplify the process of calculation of statistics. Note that correct interpreta-
tion of the output requires an understanding of the terminology used and the
underlying process of calculation, and this may best be obtained by working
through one or more examples by hand before using these tools. Spreadsheets
offer increasingly sophisticated statistical analysis functions, some examples
of which are provided in Box 53.3 for Microsoft Excel.

Box 53.3 How to use a spreadsheet (Microsoft Excel) to calculate descriptive statistics
Method 1: Using spreadsheet functions to generate the required statistics. Suppose you had obtained the following
set of data, stored within an array (block of columns and rows) of cells (A2:L6) within a spreadsheet:
Taking Microsoft Excel as an example, the following functions could be used to extract descriptive statistics from
this data set:
Descriptive statistic Example of use of functiona,b Result for the above data set
Sample size n = COUNT(A2:L6) 60

Mean = AVERAGE(A2:L6) c
4.9
Median = MEDIAN(A2:L6) 4.0
Mode = MODE(A2:L6) 3
Upper quartile = QUARTILE(A2:L6,3) d
6.0
Lower quartile = QUARTILE(A2:L6,1) 3.0
Semi-interquartile range = QUARTILE(A2: L6,3)–QUARTILE(A2:L6,1) 3.0
Upper extreme = QUARTILE(A2:L 6,4) or = MAX(A2:L 6) 11
Lower extreme = QUARTILE(A2:L 6,0) or = MIN(A2:L 6) 2
Range = MAX(A2:L6)9MIN(A2:L6) e
9.0
Variance = VAR(A2:L6) 4.464
Standard deviation = STDEV(A2:L6) 2.113
Standard error = STDEV(A2:L6)/(SQRT(COUNT(A2:L6))) f
0.273
Coefficient of variation = 100*STDEV(A2:L6)/AVERAGE(A2:L6) 43.12%
Notes:
a
Typically for Excel, in an appropriate cell, you would select the Formulas tab, then More Functions 7 Statistical 7 COUNT, then
select the input range and press return.
b
Other descriptive statistics can be calculated – these mirror those shown in Box 53.1, but for this specific data set.
c
There is no ‘MEAN’ function in Microsoft Excel.
d
The first argument within the brackets relates to the array of data, the second relates to the quartile required (consult the Help on
this function feature for further information).
e
There is no direct ‘RANGE’ function in Microsoft Excel.
f
There is no direct ‘STANDARD ERROR’ function in Microsoft Excel. The SQRT function returns a square root and the COUNT
function determines the number of filled data cells in the array.

Method 2: Using the Tools 7 Data Analysis option. This Descriptive statistics for a data set
can automatically generate a table of descriptive statistics
for the data array selected, although the data must be Column 1a,b
presented as a single row or column. This option might Mean 4.9
need to be installed for your network or personal com-
Standard error 0.27
puter before it is available to you (consult the Help feature
for details). Having entered or rearranged your data into a Median 4.0
row or column, the steps involved are as follows: Mode 3
Standard deviation 2.113
1. Select Data 7 Data Analysis.
Sample variance 4.464
2. From the Data Analysis box, select Descriptive Kurtosis 0.22
Statistics.
Skewness 0.86
3. Input your data location into the Input Range (left click Range 9.00
and hold down to highlight the column of data). Minimum 2.0
4. From the menu options, select Summary Statistics Maximum 11.0
and Confidence Level for Mean: 95%. Sum 294
5. When you click OK, you should get a new work- Count 60
sheet, with descriptive statistics and confidence Confidence level (95.0%) 0.55
limits shown. Alternatively, at step 3, you can select
Notes:
an area of your current worksheet as a data output a
These descriptive statistics are specified (and are auto-
range (select an area away from any existing content
matically presented in this order) – any others required can
as these cells would otherwise be overwritten by the be generated using Method 1.
descriptive statistics output table). b
A more descriptive heading can be added if desired – this
6. Change the format of the cells to show each number is the default.
to an appropriate number of decimal places. You may
also wish to make the columns wider so you can read
Note: instructions illustrated here may vary among the
their content.
different versions of Microsoft Office programs. Check
7. For the data set shown above, the final output table the exact functions and syntax using the Insert function
should look as shown below. menu option (fx button in some versions).

Currell, G and Dowman, T. (2009) Essential Mathematics Schmuller, J. (2013) Statistical Analysis with Excel for
and Statistics for Science, 2nd edn. John Wiley & Sons Dummies, 3rd edn. John Wiley & Sons Ltd, Hoboken.
Ltd, Chichester. Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
Dytham C. (2010) Choosing and Using Statistics: A Biolo- Oxford University Press, Oxford.
gists’ Guide, 3rd edn. Blackwell, Oxford. Taylor, J.K. and Cihon, C. (2004) Statistical Techniques for
Miller, J.N. and Miller, J.C. (2010) Statistics and Chemom- Data Analysis, 2nd edn. CRC Press, Boca Raton.
Harlow.
Press, Oxford.

Study exercises
53.1 Practise calculating descriptive statistics. Using number of data values = 12; Sample B,
the data set given in Box 53.3, calculate the fol- mean = 13.2, standard deviation 14.4, number of
lowing statistics: data values = 20. Which has the lower standard
error in absolute terms and in proportion to the
(a) range
sample mean? (Express answers to three signif-
(b) variance
icant figures.)
(c) standard deviation
(d) coefficient of variation 53.3 Compute a mean value correctly. A researcher
(e) standard error. finds that the mean vitamin concentration
in three replicate samples designated A, B
Answers (b) to (e) should be given to three sig-
and C is 3.0, 2.5 and 2.0 mg, respectively. He
nificant figures.
computes the mean vitamin concentration as
53.2 Calculate and interpret standard errors. Two 2.5 mg, but forgets that the sample sizes were
samples, A and B, gave the following descriptive 24, 37 and 6, respectively. What is the true
statistics (measured in the same units): Sam- mean vitamin concentration? (Answer to three
ple A, mean = 16.2, standard deviation = 12.7, significant figures.)

54 Choosing and using statistical tests
This chapter outlines the philosophy of hypothesis-testing statistics, indicates

Definition the steps to be taken when choosing a test, and discusses features and assump-
tions of some important tests. For details of the mechanics of tests, appropriate
Null hypothesis – a statement of ‘no
texts should be consulted (e.g. Miller and Miller, 2010). Most tests are now
effect’ in relation to the comparison
available in statistical packages for computers (see p. 436) and many in spread-
under test.
sheets (Chapter 45).
To carry out a statistical test:
1. decide what it is you wish to test (create a null hypothesis and its
alternative).
2. determine whether your data fit a standard distribution pattern;
3. select a test and apply it to your data.
Setting up a null hypothesis

(a)
mean for mean for
Hypothesis-testing statistics are used to compare the properties of samples
either with other samples or with some theory about them. For instance, you
may be interested in whether two samples can be regarded as having different
means, whether the vitamin content in different products can be regarded as
randomly distributed, or whether characteristic A of the vitamin is linearly
Frequency
related to characteristic B.
Key Point You cannot use statistics to prove any hypothesis,

but they can be used to assess how likely it is to be wrong.
(b) Statistical testing operates in what at first seems a rather perverse manner.
Suppose you think a treatment has an effect. The theory you actually test is
that it has no effect; the test tells you how improbable your data would be if
this theory were true. This ‘no effect’ theory is the null hypothesis (NH). If
your data are very improbable under the NH, then you may suppose it to be
wrong, and this would support your original idea (the ‘alternative hypothesis’).
Frequency
The concept can be illustrated by an example. Suppose two groups of subjects

were treated in different ways, and you observed a difference in the mean value
of the measured variable for the two groups. Can this be regarded as a ‘true’
difference? As Fig. 54.1 shows, it could have arisen in two ways:
1. Because of the way the subjects were allocated to treatments, i.e. all the
Dependent (measured) variable subjects liable to have high values might, by chance, have been assigned
to one group and those with low values to the other (Fig. 54.1(a)).
Fig. 54.1 Two explanations for the differ-
ence between two means. In case (a) the 2. Because of a genuine effect of the treatments, i.e. each group came from
two samples happen by chance to have a distinct frequency distribution (Fig. 54.1(b)).
come from opposite ends of the same fre-
A statistical test will indicate the probabilities of these options. The NH
quency distribution, i.e. there is no true
difference between the samples. In case
states that the two groups come from the same population (i.e. the treatment
(b) the two samples come from different effects are negligible in the context of random variation). To test this, you
frequency distributions, i.e. there is a true calculate a test statistic from the data, and compare it with tabulated critical
difference between the samples. In both values giving the probability of obtaining the observed or a more extreme result
cases, the means of the two samples are by chance (see Boxes 54.1 and 54.2). This probability is sometimes called the
the same. significance of the test.

Choosing and using statistical tests
Note that you must take into account the degrees of freedom (d.f) when
looking up critical values of most test statistics. The d.f. is related to the size(s)
of the samples studied; formulae for calculating it depend on the test being
used. Biologists normally use two-tailed tests, i.e. we have no expectation
beforehand that the treatment will have a positive or negative effect compared
to the control (in a one-tailed test we expect one particular treatment to be
bigger than the other). Be sure to use critical values for the correct type of test.
By convention, the critical probability for rejecting the NH is 5%
Definition (i.e. P = 0.05). This means we reject the NH if the observed result would have
come up by chance a maximum of one time in twenty. If the modulus of the test
Modulus – the absolute value of a num-
statistic is less than or equal to the tabulated critical value for P = 0.05, then
ber, e.g. modulus - 3.385 = 3.385.
we accept the NH and the result is said to be ‘not significant’ (NS for short). If
the modulus of the test statistic is greater than the tabulated value for P = 0.05,
then we reject the NH in favour of the alternative hypothesis that the treatments
Quoting significance – the convention had different effects and the result is ‘statistically significant’.
for quoting significance levels in text, Two types of error are possible when making a conclusion on the basis of a
tables and figures is as follows: statistical test. The first occurs if you reject the NH when it is true and the sec-
P 7 0.05 = ‘not significant’ (or NS) ond if you accept the NH when it is false. To limit the chance of the first type of
P 8 0.05 = ‘significant’ (or *) error, choose a lower probability, e.g. P = 0.01, but note that the critical value
P 8 0.01 = ‘highly significant’ (or **) of the test statistic increases when you do this and results in the probability of
the second type of error increasing. The conventional significance levels given
P 8 0.001 = ‘very highly significant’
in statistical tables (usually 0.05, 0.01, 0.001) are arbitrary. Increasing use of
(or ***).
statistical computer programs now allows the actual probability of obtaining
Thus, you might refer to a difference the calculated value of the test statistic to be quoted (e.g. P = 0.037).
in means as being ‘highly significant Note that if the NH is rejected, this does not tell you why many alterna-
(P 8 0.01)’. For this reason, the word tive explanations may be possible. Also, it is important to distinguish between
‘significant’ in its everyday meaning statistical significance and chemical relevance: identifying a statistically sig-
of ‘important’ or ‘notable’ should be nificant difference between two samples does not mean that this will carry any
used with care in scientific writing. The chemical importance.
asterisk coding shown above is most
often used as a short-form in tables and
should be explained in the table legend, Comparing data with parametric distributions
or in a footnote.
The distribution pattern of a set of data values may be chemically relevant,
but it is also of practical importance because it defines the type of statis-
tical tests that can be used. A parametric test is one that makes particular
assumptions about the mathematical nature of the population distribution from
Choosing between parametric and
which the samples were taken. If these assumptions are not true, then the
non-parametric tests – always plot
test is obviously invalid, even though it might give the answer we expect. A
your data graphically when determin-
non-parametric test does not assume that the data fit a particular pattern, but
ing whether they are suitable for par-
it may assume some characteristics of the distributions. Used in appropriate
ametric tests as this may save a lot of
circumstances, parametric tests are better able to distinguish between true but
unnecessary effort later.
marginal differences between samples than their non-parametric equivalents
(i.e. they have greater ‘power’).
The properties of the main distribution types found in chemistry are given
below, with both rules-of-thumb and more rigorous tests for deciding whether
data fit these distributions.
Binomial distributions
These apply to samples of any size from populations when data values occur
independently in only two mutually exclusive classes (e.g. type A or type B).
They describe the probability of finding the different possible combinations

0.6
(a) P = 0.1 (b) P = 0.5 (c) P = 0.7
0.5
Relative frequency
0.4
0.3
0.2
0.1
0 1 2 3 4 5 0 1 2 3 4 5 0 1 2 3 4 5
Number of individuals of type A in sample
Fig. 54.2 Examples of binomial frequency distributions with different prob-

abilities. The distributions show the expected frequency of obtaining n
individuals of type A in a sample of 5. Here P is the probability of an indi-
vidual being type A rather than type B.
of the attribute for a specified sample size k (e.g. out of ten samples, what is
the chance of eight being type A). If p is the probability of the attribute being
of type A and q the probability of it being type B, then the expected mean
Quantifying skew – the Microsoft Excel sample number of type A is kp and the standard deviation is Ukpq. Expected
SKEW function can be used to assess frequencies can be calculated using mathematical expressions (see miller
the extent of skewness in a data set. and miller, 2010). Examples of the shapes of some binomial distributions are
shown in Fig. 54.2. Note that they are symmetrical in shape for the special
case p = q = 0.5 and the greater the disparity between p and q, the more
skewed the distribution.
Some chemical examples of data likely to be distributed in binomial fash-
ion are: a drug in a pharmaceutical product; vitamin C in fruit; and the absence
or presence of a pesticide in a soil sample. To establish whether a set of data
is distributed in binomial fashion, calculate expected frequencies from prob-
ability values obtained from theory or observation, then test against observed
frequencies using a x2@test or a G-test.
Poisson distributions
Tendency towards the normal
These apply to discrete characteristics which can assume low whole num-
distribution – under certain conditions,
ber values, such as counts of events occurring in area, volume or time. The
binomial and Poisson distributions can
events should be ‘rare’ in that the mean number observed should be a small
be treated as normally distributed:
proportion of the total that could possibly be found. Also, finding one count
●● where samples from a binomial dis- should not influence the probability of finding another. The shape of Poisson
tribution are large (i.e. 7 15) and p distributions is described by only one parameter, the mean number of events
and q are close to 0.5;
observed, and has the special characteristic that the variance is equal to the
●● for Poisson distributions, if the num-
mean. The shape has a pronounced positive skewness at low mean counts,
ber of counts recorded in each out-
come is greater than about 15. but becomes more and more symmetrical as the mean number of counts
increases (Fig. 54.3).
An example of characteristics distributed in a Poisson fashion is number of
radioactive disintegrations per unit time. One of the main uses for the Poisson
distribution is to quantify errors in count data such as the number of minor
accidents in the chemical laboratory over the course of an academic year. To
decide whether data are Poisson distributed:
●● use the rule of thumb that if the coefficient of dispersion ≈1, the distribution
is likely to be Poisson;

0.4
(a) Y = 1.0
0.3
(b) Y = 2.5
Relative frequency
0.2
(c) Y = 12
0.1
0 1 2 3 4 5 0 1 2 3 4 5 6 7 8 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
Number of individuals of type A in sample
Fig. 54.3 Examples of Poisson frequency distributions differing in mean. The distributions are shown as line charts
because the independent variable (events per sample) is discrete.
●● calculate ‘expected’ frequencies from the equation for the Poisson distribu-
Definition tion and compare with actual values using a x2@test or a G-test.
Coefficient of dispersion = s 2/Y. This is It is sometimes of interest to show that data are not distributed in a Poisson
an alternative measure of dispersion to fashion, e.g. the distribution of parasite larvae in hosts. If s2/Y 7 1, the data
the coefficient of variation (p. 485). are ‘clumped’ and occur together more than would be expected by chance; if
s2/Y 6 1, the data are ‘repulsed’ and occur together less frequently than would
be expected by chance.
Normal distributions (Gaussian distributions)

These occur when random events act to produce variability in a continuous char-
acteristic (quantitative variable). This situation occurs frequently in chemistry,
so normal distributions are very useful and much used. The bell-like shape of
normal distributions is specified by the population mean and standard deviation
(Fig. 54.4): it is symmetrical and configured such that 68.27% of the data will
lie within {1 standard deviation of the mean, 95.45% within {2 standard
deviations of the mean, and 99.73% within {3 standard deviations of the mean.
Some chemistry examples of data likely to be distributed in a normal fash-
ion are: pH of natural waters; melting point of a solid compound. To check
whether data come from a normal distribution, you can:
●● use the rule of thumb that the distribution should be symmetrical and that
nearly all the data should fall within {3s of the mean and about two-thirds
within {1s of the mean;
●● plot the distribution on normal probability graph paper. If the distribution is
normal, the data will tend to follow a straight line (see Fig. 54.5). Deviations
from linearity reveal skewness and/or kurtosis (see pp. 486), the significance
of which can be tested statistically (see Miller and Miller, 2010);

(a)
Frequency
(b) (c)
Dependent variable
Fig. 54.4 Examples of normal frequency distributions differing in mean and standard deviation. The horizontal bars
represent population standard deviations for the curves, increasing from (a) to (c). Vertical dashed lines are population
means, while vertical solid lines show positions of values {1, 2 and 3 standard deviations from the means.
●● use a suitable statistical computer program to generate predicted normal

curves from the Y and s values of your sample(s). These can be compared
98
visually with the actual distribution of data and can be used to give ‘expected’
95 values for a x2@test or a G-test.
90 The wide availability of tests based on the normal distribution and their rel-
ative simplicity means that you may wish to transform your data to make them
80 more like a normal distribution. Table 54.1 provides transformations that might
Cumulative frequency (%)
70
be applied. The transformed data should be tested for normality as described
Y+s above before proceeding – do not forget that you may need to check that trans-
60
formed variances are homogeneous for certain tests (see below).
50
A very important theorem in statistics, the Central Limit Theorem, states
40
that, as sample size increases, the distribution of a series of means from any
30
Y frequency distribution will become normally distributed. This fact can be used
20 to devise an experimental or sampling strategy that ensures that data are nor-
mally distributed, i.e. using means of samples as if they were primary data.
10
5
Choosing a suitable statistical test
Y–s
Comparing location (e.g. means)
2
If you can assume that your data are normally distributed, the main test for
2 3 4 5 6 7 8 9 10 comparing two means from independent samples is Student’s t-test (see
Upper class limit Boxes 54.1 – 54.3, and Table 54.2). This assumes that the variances of the
Fig. 54.5 Example of a normal probability
plot. The plotted points are from a small Table 54.1 Suggested transformations altering different types of frequency
data set where the mean Y = 6.93 and the distribution to the normal type. To use, modify data by the formula shown;
standard deviation s = 1.895. Note that then examine effects with the tests described on pp. 496–497; 501–503.
values corresponding to 0% and 100%
cumulative frequency cannot be used. The Type of data; distribution suspected Suggested transformation(s)
straight line is that predicted for a normal
Proportions (including percentages); arcsine Ux (also called the angular
distribution with Y = 6.93 and s = 1.895. binomial transformation)
This is plotted by calculating the expected
positions of points for Y {s. Since 68.3% of Scores; Poisson Ux or U(x + 1/2) if zero values present
the distribution falls within these bounds, Measurements; negatively skewed x2, x3, x4, etc. (in order of increasing
the relevant points on the cumulative fre- strength)
quency scale are 50{34.15%; thus this line
Measurements; positively skewed 1/Ux, Ux, In x, 1/x (in order of increasing
was drawn using the points (4.495, 15.85)
strength)
and (8.285, 84.15) as indicated on the plot.

Box 54.1 How to carry out a t-test
The t-test was devised by a statistician who used the this table is designed for ‘two-tailed’ tests, i.e. where the
pen-name ‘Student’, so you may see it referred to as Stu- treatment or sampling strategy could have resulted in
dent’s t-test. It is used when you wish to decide whether either an increase or a decrease in the measured values.
two samples come from the same population or from These are the most likely situations you will deal with in
different ones (Fig. 54.1). The samples might have been biology.
obtained by observation, or by applying two different Examine Table 54.2 and note the following.
treatments to an originally homogeneous population
●● The larger the size of the samples (i.e. the greater
(Chapter 5).
the ‘degrees of freedom’), the smaller t needs to be
The null hypothesis (NH) is that the two groups can be
to exceed the critical value at a given significance
represented as samples from the same overlying popu-
level.
lation (Fig. 54.1(a)). If, as a result of the test, you accept
this hypothesis, you can say that there is no significant ●● The lower the probability, the greater t needs to be to
difference between the group means. exceed the critical value.
The alternative hypothesis is that the two groups come
from different populations (Fig. 54.1(b)). By rejecting the The mechanics of the test
NH as a result of the test, you can accept the alternative A calculator that can work out means and standard devi-
hypothesis and say that there is a significant difference ations is helpful.
between the sample means, or, if an experiment were
carried out, that the two treatments affected the samples 1. Work out the sample means Y1 and Y2 and calculate
differently. the difference between them.
How can you decide between these two hypotheses? 2. Work out the sample standard deviations S1 and S2.
On the basis of certain assumptions (see below), and (Note: if your calculator offers a choice, choose the
some relatively simple calculations, you can work out the ‘n - 1’ option for calculating s – see pp. 485).
probability that the samples came from the same popula-
tion. If this probability is very low, then you can reasona- 3. Work out the sample standard errors Se1 = s1/2n1
bly reject the NH in favour of the alternative hypothesis, and Se2 = s2/2n2; now square each, add the squares
and if it is high, you will accept the NH. together, then take the positive square root of this (n1
To find out the probability that the observed differ- and n2 are the respective sample sizes, which may, or
ence between sample means arose by chance, you must may not, be equal).
first calculate a ‘t value’ for the two samples in question.
4. Calculate t from the formula:
Some computer programs (e.g. Minitab) provide this
probability as part of the output, otherwise you can look
Y1 - Y2
up statistical tables (e.g. Table 54.2). These tables show t =
‘critical values’ – the borders between probability levels. 2 1 (SE1)2 2 + 1 SE2)2 2
If your value of t equals or exceeds the critical value for
probability P, you can reject the NH at this probability The value of t can be negative or positive, depending
(‘level of significance’). on the values of the means; this does not matter and
Note that: you should compare the modulus (absolute value) of
●● for a given difference in the means of the two samples, t with the values in tables.
the value of t will get larger the smaller the scatter 5. Work out the degrees of freedom = (n1 - 1) +
within each data set; and (n2 - 1).
●● for a given scatter of the data, the value of t will get 6. Compare the t value with the appropriate critical
larger the greater the difference between the means. value (see, e.g., Table 54.2) and decide on the signifi-
So, at what probability should you reject the NH? Nor- cance of your finding (see p. 491).
mally, the threshold is arbitrarily set at 5% – you quite Box 54.2 provides a worked example – use this to check
often see descriptions like ‘the sample means were sig- that you understand the above procedures.
nificantly different (P 6 0.05)’. At this ‘significance level’
there is still up to a 5% chance of the t value arising by Assumptions that must be met before using the test
chance, so about 1 in 20 times, on average, the conclu-
The most important assumptions are as follows.
sion will be wrong. If P turns out to be lower, then this
kind of error is much less likely. ●● The two samples are independent and randomly drawn
Tabulated probability levels are generally given for 5%, (or, if not, drawn in a way that does not create bias). The
1% and 0.1% significance levels (see Table 54.2). Note that test assumes that the samples are quite large.

●● The underlying distribution of each sample is nor- two-thirds within one standard deviation of the mean
mal. This can be tested with a special statistical test, (see p. 494).
but a rule of thumb is that a frequency distribution
of the data should be (a) symmetrical about the ●● The two samples should have uniform variances. This
mean and (b) nearly all of the data should be within again can be tested (by an F-test), but may be obvious
three standard deviations of the mean and about from inspection of the two standard deviations.
Box 54.2 Worked example of a t-test
Suppose the following data were obtained in an experi- -0.8666 -0.8666

4. t = =
ment (the units are not relevant): 2
2(0.202 348 + 0.193 934 ) 2 0.280 277
Control: 6.6, 5.5, 6.8, 5.8, 6.1, 5.9 = -3.09.
Treatment: 6.3, 7.2, 6.5, 7.1, 7.5, 7.3
5. d.f. = (5 + 5) = 10.
Using the steps outlined in Box 54.1, the following
6. Looking at Table 54.2, we see that the modulus of
values are obtained (denoting control with subscript 1,
this t value exceeds the tabulated value for P = 0.05
treatment with subscript 2):
at 10 degrees of freedom ( = 2.23). We therefore
1. Y1 = 6.1167; = Y2 = 6.9833: difference between reject the NH, and conclude that the means are dif-
means = Y1 - Y2 = - 0.8666. ferent at the 5% level of significance. If the modulus
of t had been 8 2.23, we would have accepted the
2. s1 = 0.495 65; s2 = 0.475 04.
NH. If the modulus of t had been 73.17, we could
3. SE1 = 0.495 65/2.449 49 = 0.202 348 have concluded that the means are different at the
SE2 = 0.475 04/2.449 49 = 0.193 934. 1% level of significance.
Table 54.2 Critical values of Student’s t statistic (for two-tailed tests).

Reject the null hypothesis at probability P if your calculated t value
equals or exceeds the value shown for the appropriate degrees of
freedom = (n1 - 1) + (n2 - 1).
Degrees of Critical values Critical values Critical values

freedom for P = 0.05 for P = 0.01 for P = 0.001
1 12.71 63.66 636.62

2 4.30 9.92 31.60
3 3.18 5.84 12.94
4 2.78 4.60 8.61
5 2.57 4.03 6.86
6 2.45 3.71 5.96
7 2.36 3.50 5.40
8 2.31 3.36 5.04
9 2.26 3.25 4.78
10 2.23 3.17 4.59
12 2.18 3.06 4.32
14 2.14 2.98 4.14
16 2.12 2.92 4.02
20 2.09 2.85 3.85
25 2.06 2.79 3.72
30 2.04 2.75 3.65
40 2.02 2.70 3.55
60 2.00 2.66 3.46
120 1.98 2.62 3.37
∞ 1.96 2.58 3.29

data sets are homogeneous. Tests based on the t-distribution are also available
Definition for comparing means of paired data or for comparing a sample mean with a
chosen value.
Homogeneous variance – uniform
When comparing means of two or more samples, analysis of variance
(but not necessarily identical) variance
(ANOVA) is a very useful technique. This method also assumes data are nor-
of the dependent variable across the
range of the independent variable. The mally distributed and that the variances of the samples are homogeneous. The
term homoscedastic is also used in this samples must also be independent (e.g. not subsamples). The test statistic cal-
sense. The opposite of homogeneous is culated is denoted F and it has two different degrees of freedom related to the
heterogeneous ( = heteroscedastic). number of means tested and the pooled number of replicates per mean. The
nested types of ANOVA are useful to determine the relative importance of
different sources of variability in your data. Two-way and multi-way ANOVAs
are useful for studying interactions between treatments.
Understanding ‘degrees of freedom’ – For data satisfying the ANOVA requirements, the least significant dif-
this depends on the number of values in ference (LSD) is useful for making planned comparisons among several
the data set analysed, and the method means (see Miller and Miller, 2010). Any two means that differ by more
of calculation depends on the statistical than the LSD will be significantly different. The LSD is useful for showing
test being used. It relates to the num- on graphs.
ber of observations that are free to vary The chief non-parametric tests for comparing the locations of two sam-
before the remaining quantities for a ples are the Mann–Whitney U-test and the Kolmogorov–Smirnov test. The
data set can be determined. former assumes that the frequency distributions of the samples are similar,
whereas the latter makes no such assumption. In both cases the sample’s size
must be ⩾ 4 and for the Kolmogorov–Smirnov test the samples must have
equal sizes. In the Kolmogorov–Smirnov test, significant differences found
with the test could be caused by differences in location or shape of the dis-
Checking the assumptions of a test –
tribution, or both.
always acquaint yourself with the
Suitable non-parametric comparisons of location for paired data (sample
assumptions of a test. If necessary, test
size ⩾ 6) include Wilcoxon’s signed rank test, which is used for quantitative
them before using the test.
data and assumes that the distributions have similar shape. Dixon and Mood’s
sign test can be used for paired data scores where one variable is recorded as
‘greater than’ or ‘better than’ the other.
Non-parametric comparisons of location for three or more samples include
the Kruskal–Wallis H-test. Here, the number of samples is without limit and
they can be unequal in size, but again the underlying distributions are assumed
to be similar. The Friedman S-test operates with a maximum of five samples
and data must conform to a randomised block design. The underlying distribu-
tions of the samples are assumed to be similar.
Comparing dispersions (e.g. variances)

If you wish to compare the variances of two sets of data that are normally dis-
tributed, use the F-test. For comparing more than two samples, it may be suffi-
cient to use the Fmax@test, on the highest and lowest variances. The Scheffé–Box
(log-ANOVA) test is recommended for testing the significance of differences
between several variances. Non-parametric tests exist but are not widely avail-
able: you may need to transform the data and use a test based on the normal
distribution.
Confidence limits for statistics other Comparing proportion data

than the mean – consult an advanced When comparing proportions between two small groups (e.g. whether 3/10 is
statistical text (e.g. Miller and Miller, significantly different from 5/10), you can use probability tables such as those
2010) if you wish to indicate the reliabil- of Finney et al. (1963) or calculate probabilities from formulae; however, this
ity of estimates of, for example, popu- can be tedious for large sample sizes. Certain proportions can be transformed
lation variances. so that their distribution becomes normal.

Placing confidence limits on an estimate of a population

parameter
On many occasions, a sample statistic is used to provide an estimate of a pop-
ulation parameter, and it is often useful to indicate the reliability of such an
estimate. This can be done by putting confidence limits on the sample statistic,
i.e. by specifying an interval around the statistic within which you are confident
that the true value (the population parameter) is likely to fall, at a specified
level of probability. The most common application is to place confidence limits
on the mean of a sample taken from a population of normally distributed data
values. In practice, you determine a confidence factor for a particular level of
probability which is added to and subtracted from the sample mean (Y) to give
the upper confidence limit and lower confidence limit respectively. These are
calculated as:
Y + (tP[n - 1] * SE) for the upper limit and

Y - (tP[n - 1] * SE) for the lower limit[54.2]
where tP[n - 1] is the tabulated critical value of Student’s t-statistic for a

two-tailed test with n - 1 degrees of freedom at a specified probability level
(P) and SE is the standard error of the sample mean (p. 485). The 95% con-
fidence limits (i.e. P = 0.05) tells you that, on average, 95 times out of 100,
the interval between the upper and lower limits will contain the true (popula-
tion) value. Confidence limits are often shown as ‘error bars’ for individual
sample means plotted in graphical form. Figure 54.6 illustrates how this is
applied to plotted curves and histograms (note that this can be carried out for
data series within a Microsoft Excel graph (chart) using the Layout tab and
Error Bars menu).
(a) (b) Correlation and regression

upper confidence
limit These methods are used when testing the relationship between data values for
confidence factor two variables. Correlation is used to measure the extent to which changes in the
two sets of data values occur together in a linear manner. If one variable can be
confidence
interval
sample mean assumed to be dependent upon the other (i.e. a change in X causes a particular
change in Y), then regression techniques can be used to provide a mathematical
confidence factor description of the underlying relationship between the variables, e.g. to find a
lower confidence line of best fit for a data series. If there is no a priori reason to assume depend-
limit
ency, then correlation methods alone are appropriate.
Fig. 54.6 Graphical representation of confi- A correlation coefficient measures the strength of the linear relationship
dence limits as ‘error bars’ for (a) a sample between two variables, but does not describe the relationship. The coefficient
mean in a plotted curve, where both upper is expressed as a number between -1 and +1: a positive coefficient indicates
and lower limits are shown; and (b) a sam- a direct relationship, where the two variables change in the same direction,
ple mean in a histogram, where, by con- while a negative coefficient indicates an inverse relationship, where one vari-
vention, only the upper value is shown. able decreases as the other increases (Fig. 54.7). The nearer the coefficient is
For data that are assumed to be symmetri-
to -1 or +1, the stronger the linear relationship between the variables, i.e. the
cally distributed, such representations are
less ‘scatter’ there would be about a straight line of best fit (note that this
often used in preference to the ‘box and
whisker’ plot shown on p. 485. Note that does not necessarily imply that one variable is dependent upon the other). A
SE is an alternative way of representing coefficient of 0 implies that the two variables show no linear association and
sample imprecision/error (e.g. Fig. 49.1). therefore the closer the correlation coefficient is to zero, the weaker the linear
See p. 458 for how to add error bars to relationship. The importance of graphing data is shown by the case illustrated
Microsoft Excel graphs. in Fig. 54.7(d).

Pearson’s product moment correlation coefficient (r) is the most com-

(a)
monly used statistic for testing correlations. The test is valid only if both var-
iables are normally distributed. Statistical tests can be used to decide whether
the correlation is significant, e.g. using a one-sample t-test to see whether r is
significantly different from zero, based on the following equation:
y
t = r , U[(1 - r 2) , (n - 2)][54.3]
at n - 2 degrees of freedom, where n is the number of paired observa-

tions. If one or both variables are not normally distributed, then you should
x
calculate an alternative non-parametric coefficient, e.g. Spearman’s coeffi-
(b) cient of rank correlation (rS) or Kendall’s coefficient of rank correlation (t).
These require the two sets of data to be ranked separately, and the calculation
can be complex if there are tied (equal) ranks. Spearman’s coefficient is said
to be better if there is any uncertainty about the reliability of closely ranked
data values.
y
If underlying theory or empirical graphical analysis indicates a linear
relationship between a dependent and an independent variable, then linear
regression can be used to estimate the mathematical equation that links the
two variables. Model I linear regression is the standard approach, and is
x available within general-purpose software programs such as Microsoft Excel
(Box 54.3), and on some scientific calculators. It is suitable for experiments
(c)
where a dependent variable Y varies with an error-free independent variable
X in accordance with the relationship Y = a + bX + eY, where eY repre-
sents the residual (error) variability in the Y variable. For example, this
relationship might apply in a laboratory procedure where you have care-
y fully controlled the independent variable and the X values can be assumed
to have zero error (e.g. in a calibration curve, see Chapter 48, or in a time
course experiment where measurements are made at exact time points). The
regression analysis gives estimates for a and b (equivalent to the slope and
intercept of the line of best fit, p. 472): computer-based programs usually
x
provide additional features, e.g. residual values for Y (eY), estimated errors
(d) for a and b, predicted values of Y along with graphical plots of the line of
best fit (the trend line) and the residual values. In order for the model to be
valid, the residual (error) values should be normally distributed around the
trend line and their variance should be uniform (homogeneous), i.e. there
should be a similar scatter of data points around the trend line along the
y
x-axis (independent variable).
If the relationship is not linear, try a transformation. For example, this
is commonly done in physical chemistry (kinetics). However, you should be
aware that the transformation of data to give a straight line can lead to errors
x
when carrying out linear regression analysis: take care to ensure that (a) the
assumptions listed in the previous paragraph are valid for the transformed data
Fig. 54.7 Examples of correlation. The linear set, and (b) the data points are evenly distributed throughout the range of the
regression line is shown. In (a) and (b), the independent variable. If these criteria cannot be met, non-linear regression may
correlation between x and y is good: for (a) be a better approach, but for this you will require a suitable computer program,
there is a positive correlation and the cor-
e.g. GraphPad Prism.
relation coefficient, r, would be close to 1;
The strength of the relationship between Y and X in model I linear regres-
for (b) there is a negative correlation and the
correlation coefficient would be close to - 1. sion is best estimated by the coefficient of determination (r 2 or R2), which
In (c) there is a weak positive correlation and is equivalent to the square of the Pearson correlation coefficient. The coef-
r would be close to 0. In (d) the correlation ficient of determination varies between 0 and +1 and provides a measure of
coefficient may be quite large, but the choice the goodness of fit of the Y data to the regression line: the closer the value
of linear regression is clearly inappropriate. is to 1, the better the fit. In effect, r 2 represents the fraction of the variance

Using more advanced types of in Y that can be accounted for by the regression equation. Conversely, if you
regression – these include the following. subtract this value from 1, you will obtain the residual (error) component,
i.e. the fraction of the variance in Y that cannot be explained by the line of
●● Model II linear regression, which best fit. Multiplying the values by 100 allows you to express these fractions
applies to situations where a in percentage terms.
dependent variable Y varies with an
independent variable X, and where
both variables may have error terms
associated with them. Using computers to calculate hypothesis-testing statistics
●● Multiple regression, which applies As with the calculation of descriptive statistics (p. 487), specialist statistical
when there is a relationship between
packages such as SPSS and MINITAB can be used to simplify the calculation
a dependent variable and two or
more independent variables.
of hypothesis-testing statistics. The correct use of the software and interpreta-
●● Non-linear regression, which extends tion of the output requires an understanding of relevant terminology and of the
the principles of linear regression to fundamental principles governing the test, which is probably best obtained by
a wide range of functions. Techni- working through one or more examples by hand before using these tools (e.g.
cally, this method is more appropri- Box 54.2). Spreadsheets offer increasingly sophisticated statistical analysis
ate than trans-forming data to allow functions, three examples of which are provided in Box 54.3.
linear regression.
Advanced statistics books should be

consulted for details of these methods,
which may also be offered by some sta-
tistical computer programs.
Example If a regression analysis

gives a value for r 2 of 0.75
(i.e. r = 0.84), then 75% of the
v
ariance in Y can be explained by the
trend line, with 1 - r 2 = 0.25 (25%)
remaining as unexplained (residual)
variation.
Box 54.3 Using a spreadsheet (Microsoft Excel) to calculate hypothesis-testing statistics
Presented below are three examples of the use of a spreadsheet to investigate hypotheses about specific data sets.
In each case, there is a brief description of the problem, a table showing the data analysed, an outline of the Excel
commands used to carry out the analysis and an annotated table of results from the spreadsheet (you can use the
same data to check your ability to carry out the procedures).
Example 1: A t-test
As part of a project, a student measured zinc by both EDTA titration and by AAS and obtained the following results.
Zinc concentration (mg mL - 1).
Replicate 1 2 3 4 5 6 7 8 Mean Variance
EDTA 2.342 2.256 2.521 2.523 2.943 2.481 2.601 2.449 2.515 0.042
AAS 2.658 2.791 2.731 2.402 3.041 2.668 2.823 2.509 2.703 0.038
(continued)

The student proposed the null hypothesis that there was The analytical chemist wanted to know whether the
no difference between the two means and tested this observed differences were statistically significant, so
using a t-test, and had evidence from other studies that he carried out an ANOVA test, assuming the samples
the zinc concentration of replicate flasks were normally were normally distributed and the variances in the
distributed. The student had also established, by calcula- three populations were homogeneous. He selected
tion, that the assumption that the populations had homo- Data 7 Data Analysis 7 Anova: Single Factor, then
geneous variances was likely to be valid. She selected enter the data ranges, select Grouped By: Rows and
Data 7 Data Analysis 7 t@Test: Two-Sample Assuming set Alpha to 0.05 ( = P). This returned a table of data as
Equal Variances, then enter data ranges as above and then shown below.
enter 0 for Hypothesised Mean Difference and 0.05 for
Alpha ( = P). This returned a table of data as shown below.
ANOVA: single factor
t-Test: two-sample assuming equal variances SUMMARY
Variable 1 Variable 2 Groups Count Sum Average Variance
Mean 2.515 2.703 A 6 3.317 0.552 833 0.000189

Variance 0.042 0.038 B 6 3.243 0.540 5 0.000 221
Observations 8 8 C 6 3.363 0.560 5 8.51E-05
Pooled variance 0.040 D 6 3.242 0.540 333 3.95E-05
Hypothesised mean 0
difference
ANOVA
df 14
t Stat - 1.881 Source of SS df MS F P-value F crit
variation
P (T 8 t) one-tail 0.040
t Critical one-tail 1.761 Between 0.001 761 3 0.000 587 4.397 856 0.015669 3.098391
P (T 8 t) two-tail 0.081 groups
t Critical two-tail 2.145 Within 0.002 669 20 0.000133
groups
Total 0.00 443 23
The value of t obtained was - 1.881 (row 7 ‘t Stat’) and
the probability of obtaining this value for a two-tailed test
(row 10) was 0.081 (or 8.1%), so the student was able to The F value calculated was 4.397 856. This comforta-
accept the null hypothesis and conclude that zinc could bly exceeds the stated critical value (Fcrit) of 3.098 391
be measured by both techniques and obtain acceptable and the probability of obtaining this result by chance
results i.e. no significant difference between the two ana- (P-value) was calculated as 0.015669 (1.57% to three
lytical techniques. significant figures), hence the chemist was able to
reject the null hypothesis and conclude that there was
Example 2: An ANOVA test a significant difference in average pesticide concen-
An analytical chemist made six replicate measurements tration between the four batches, since P 6 0.05. Such
of four different batches (A–D) of a pesticide in four soil a finding might lead on to an investigation into why
extracts, obtaining the following data. there was batch variation, e.g. had they been stored
differently?
pesticide (ng mL-1 )
Batch \ 1 2 3 4 5 6 Mean Variance Example 3: Testing the significance of a correlation

replicate
A researcher wanted to know whether there was any
A 0.562 0.541 0.576 0.545 0.542 0.551 0.552 833 0.000189 correlation between the levels of tar and nicotine in
B 0.531 0.557 0.537 0.521 0.559 0.538 0.540 500 0.000221 cigarettes. The researcher made measurements of both
C 0.572 0.568 0.551 0.549 0.564 0.559 0.560 500 0.000085 consitituents for 10 different brands and obtained the fol-
lowing results.
D 0.532 0.548 0.541 0.538 0.547 0.536 0.540 333 0.000039

Tar and nicotine content (mg g -1) of cigarettes function PEARSON (array1, array 2) to obtain a value
of +0.950260385 for the Pearson’s product moment cor-
Brand Tar Nicotine relation coefficient r, specifying the tar content data
as array1 and the nicotine content data as array2 (the
1 12.6 0.92 Excel CORREL function can also be used to carry out
2 8.5 0.58 the same task). The researcher then used a spread-
3 15.7 1.14 sheet to calculate the t statistic (p. 495) for this r value,
4 32.5 2.16 using the formula for eqn [54.3]. The calculated value
of t was 8.6296, with 8 degrees of freedom. The criti-
5 14.5 1.03
cal value from tables (e.g. Table 54.2) at P = 0.001 is
6 3.1 0.17 5.04, so the researcher concluded that there was a very
7 18.9 1.32 highly significant positive correlation between the two
8 7.7 0.98 constituents.
9 10.7 1.13
10 6.2 0.55 Note: instructions illustrated here may vary among the
different versions of Microsoft Office programs. Check the
The researcher assumed that both variables were exact functions and syntax using the Insert function menu
normally distributed and used the Microsoft Excel option (fx button in some versions).

Finney, D.J., Latscha, R., Bennett, B.M. and Hsu, P. Schmuller, J. (2013) Statistical Analysis with Excel
(1963) Tables for Testing Significance in a 2 * 2 Table. for Dummies, 3rd edn. John Wiley & Sons Ltd. Hoboken.
Cambridge University Press, Cambridge. Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
Leardi, R. (2009) Experimental design in chemistry: A tuto- Oxford University Press, Oxford.
rial. Analytica Chimica Acta, 652, 161–172. Taylor, J.K. and Cihon, C. (2004) Statistical Techniques for
Miller, J.N. and Miller, J.C. (2010) Statistics and Chemo- Data Analysis, 2nd edn. CRC Press, Boca Raton.
metrics for Analytical Chemistry, 6th edn. Prentice Hall,
Harlow.
Press, Oxford.
Study Exercises
54.1 Calculate 95% confidence limits. What are 54.2 Practise using a t-test. chemistry student meas-
the 95% confidence limits of a sample with a ures the paracetamol concentration (mg g -1)
mean = 24.7, standard deviation = 6.8 and in tablets by two different analytical meth-
number of data values = 16? (Express your ods. Seven tablets from different batches were
answer to three significant figures.) analysed to see whether the results obtained by

the two methods differed. Carry out a t-test on ANOVA

the data and draw appropriate conclusions.
df SS MS F Significance F
-1
Paracetamol concentration (mg g ) Regression 1 607.406 285 7 607.4063 133.3954 0.000320975
Residual 4 18.21371429 4.553 429
Method 1 7.5 8.1 7.6 6.2 7.5 7.8 8.9
Total 5 625.62
Method 2 5.6 7.5 8.2 6.7 3.5 6.5 5.9
54.3 Interpret the output from Microsoft Excel linear

Standard
regression analysis. The following output rep- Coefficients error t Stat P-value
resents a regression analysis for an experiment
measuring the uptake of an amino acid by a cell Intercept 1.171428 571 1.544386 367 0.758 507 0.490383
suspension (in pmol cell-1) against time (in min- X variable 1 2.945714286 0.255 047 014 11.549 69 0.000321
utes). Based on this output, what is the form and
strength of the underlying linear relationship?
(Express the coefficients to three significant
figures.)
Output from Excel spreadsheet linear regression analysis

Summary output
Regression statistics
Multiple R 0.985 335 951
R square 0.970 886 937

Adjusted R square 0.963 608 672
Standard error 2.133876 419
Observations 6

55 Drawing chemical structures
Drawing the structure of a chemical compound is probably one of the first basic
requirements of any chemist. It requires knowledge of the chemical composi-
tion of the structure to be drawn, an understanding of the type of bonding, and
frequently a mental visualisation of the arrangement of atoms (or ions). Once
this has been assimilated it is not uncommon to draw a representation of the
structure on paper. What is often lacking is the realisation that the molecule
H should be represented in three dimensions. To some extent it is possible to
represent a three-dimensional chemical structure on a piece of paper. Fig. 55.1
C
shows the structure of methane, CH4, where standard symbols e.g. the hatched
H H line, are used to imply a direction of the bond, and one that is different to, for
H
example, the solid line. This simple notation is commonly used to give a mol-
Fig. 55.1 Methane. ecule the perception of three-dimensionality.
In co-ordination chemistry, a different approach is used; for example, in the
structure for hexaamminecobaltate complex, [Co(NH3)6]3+ , there are several
NH3
alternative approaches. The simplest way to express this structure is shown in
H3N NH3
Co
Fig. 55.2.
H3N NH3 The general points to remember are as follows:
NH3
●● Always draw chemical structures in ink (pencil fades with time).
Fig. 55.2 Structure of hexaammine-cobaltate. ●● Always ensure that the chemical structure is drawn large enough, so that no
ambiguity is possible.
●● If drawing by hand, ensure that each atom is clearly identified. This may
require the use of coloured pens.
●● Try to keep the structure as simple as possible, highlighting only the key
features.
●● Any text should appear in a clear script (by hand or word processed).
●● When possible, it is advisable to use computer software packages to generate
chemical structures.
●● Make sure no confusion is possible between different letters of the alphabet
representing elements in the Periodic Table.
●● If necessary, show the number of electrons (pairs or individual electrons)
clearly. Remember, a . (full stop) may be mistaken for a mark in the paper.
●● It may be necessary to indicate the structural formula of a molecule, e.g. in
isomerism.
Selected examples of drawing chemical structures

Organic structures
Organic molecules vary from the simplest structures containing a few atoms
to the most complex molecules comprising hundreds (see Chapter 57). Chem-
ists have devised many methods of unambiguously communicating struc-
tures which illustrate the features under discussion. Most organic chemistry
is based on the reactions of functional groups, e.g. alcohol, ester, acid, amine
with the carbon skeleton making no contribution to the chemistry. This has

Drawing chemical structures
Table 55.1 Common abbreviations for the carbon skeleton
Abbreviation Carbon skeleton Formula
Me Methyl CH3 ¬
Et Ethyl CH3CH2 ¬
Pr Propyl CH3CH2CH2 ¬
Pri Iso-propyl (CH3)2CH ¬

CH2 OH Bu Butyl CH3CH2CH2CH2 ¬
H3C CH2 OH
Bui Iso-butyl (CH3)2CHCH2 ¬
But Tert-butyl (CH3)3C ¬

CH3
CH OH OH Ph Phenyl C6H5 ¬
H3C CH2
Ac Acetyl (ethanoyl) CH3CO ¬
Ts p-Toluenesulphonyl (tosyl) CH3C6H5SO2 ¬
O O
CH2 C
H3C CH2 CH3
led to the development of two common methods of depicting organic struc-

H tures: (a) abbreviated formulae and (b) bond–line structures, which avoid the
CH C complexity and irrelevance of writing out all the C ¬ H and C ¬ C bonds in
H3C CH O O
a structural formula.
(a) (a) Abbreviated formulae The carbon skeleton, usually a saturated hydro-
carbon chain or mono-substituted benzene ring, is depicted by a two-letter
Fig. 55.3a Examples of bond-line formulae.
combination as shown in Table 55.1. All other functional groups are shown
normally.
Br CH3 Simple illustrative examples of this system are bromomethane
Br
C OH (MeBr), 2-propanol (PriOH), 2-chloro-2-methylpropane (ButCl), nitroben-
CH3CH2 CH2OH
zene (PhNO2) and N-phenylethanamide (PhNHAc).
(b) (b) Bond-line formulae This is one of the most common systems in use
today and is compatible with computer-based structure drawing pro-
Fig. 55.3b Stereochemical representation
using bond-line formulae.
grams. The carbon skeleton is represented by a series of lines showing
the bonds between the saturated carbon atoms but the hydrogen atoms
and their bonds to carbon are not shown. All other atoms and multiple
2- bonds are shown. These ideas are illustrated by the following examples
O
(Fig. 55.3(a)) and stereochemistry can be shown by the usual ‘wedge’
C
O O bonds (Fig. 55.3(b)).
(a)
Lewis structures
2- 2- 2- When drawing Lewis structures it is important to show the position and
O O O
number of electrons. For example, the Lewis structure for CO2- 3 is shown in
C C C
O O O O O O Fig. 55.4(a). Note the position of the double bond. Also, both carbon and oxy-
gen obey the octet rule, i.e. the number of electrons around each atom adds up
(b) to eight (a single bond is composed of two electrons, a double bond of four
2- electrons). Lewis structures sometimes give rise to canonical forms. The three
O
possible canonical forms for CO2- 3 are shown in Fig. 55.4(b). It is noted that
C as the position of the double bond moves the number of lone pairs of electrons
O O changes. In reality, experimental evidence indicates that the C ¬ O bond in
carbonate is composed of neither single nor double bonds but is intermediate
(c)
in bond length and strength. A more appropriate method of representing this
Fig. 55.4 Lewis diagrams for CO2-
3 . structure is by delocalisation of bonding electrons (Fig. 55.4(c)).

HCP CCP Ionic structures

In solid-state chemistry it is often necessary to draw a ‘unit cell’. The most
A C
commonly found are the hexagonal close-packed (HCP) crystal structure
and cubic close-packed (CCP) crystal structure. In the HCP structure atoms
are arranged in an ABABA repeating pattern, while in the CCP structure the
B B
arrangement is an ABCABCA pattern. In both cases it is difficult to represent
the structures without resorting to the drawing of circles. Fig. 55.5 shows the
A A close-packing arrangement for both the HCP and CCP crystal structures. In
the HCP structure the first and third layers of atoms are orientated in the same
direction (directly above one another) while in the CCP structure, the first and
(a) (b)
third layers do not coincide, i.e. no atom in the third layer is directly above an
Fig. 55.5 Close-packing arrangement for atom in the first layer.
(a) hexagonal close packing (ABABA etc.) It is usually not recommended to attempt drawing ionic structures of
and (b) cubic close packing (ABCABCA greater complexity without resorting to a specialised computer-based drawing
etc.). package.
Key Point It is often important when drawing chemical struc-

tures to impart some structural identity.
Atomic orbitals
You may need to draw a visualisation of atomic orbitals, usually the s, p and
d orbitals. This can be simplified by the use of Cartesian co-ordinates which
allow a three-dimensional representation on paper. This is neither easy to
replicate or often necessary. A simplified approach is, for example, to replace
the spherical s orbital with a circle (Fig. 55.6). Similarly, the three p orbitals
y can be represented in two dimensions by the use of correct labelling of the
z axes (Fig. 55.6). Finally, the same approach can be replicated on the five d
x orbitals (Fig. 55.6). It is worthwhile remembering that the dxy, dxz and dyz
orbitals do not reside on the axes (x, y or z), but in the plane of their respec-
S tive axes. In addition, the dx2 - y2 orbital occupies the x- and y-axes and the dz2
orbital occupies the z-axis.
y z y
Electronic configuration
x x x
It is often difficult to recollect the order of filling of the electronic structures
of atoms of different elements. Fig. 55.7 shows the usual order of filling of
py pz px
the orbitals of an atom according to the Aufbau principle (lowest energy first).
However, a simple mnemonic exists to facilitate the correct order for each
y z y element in the Periodic Table (Fig. 55.8). In order to use the mnemonic all you
need to remember is that:
x x z
●● s orbitals can have up to 2 e -
dxy dxz dyz ●● p orbitals can have up to 6 e -
y z ●● d orbitals can have up to 10 e -
x x
●● f orbitals can have up to 14 e - .
Then, it is simply a case of addition. In general terms, the order of electron
dx 2-y 2 pz 2 filling is as follows (simply translating the mnemonic into an order):
1s 2s 2p 3s 3p 4s 3d 4p 5s 4d 5p 6s 4f 5d 6p 7s . . .
Fig. 55.6 Atomic orbitals.

1s s p d f
2s 2p 6d
5f
3s 3p 3d 7s 6p
4d
4s 4p 4d 4f 4f
5s 5p 5d 5f 5g 6s
5p
4d
6s 6p 6d 6f 6g 6h
5s
7s …
4p
increasing energy
3d
Fig. 55.8 Mnemonic for electronic configu- 4s
ration determination based on the Aufbau
3p
principle.
3s
2p
2s
1s
Fig. 55.7 Electronic configuration.
For example, the atomic number of calcium is 20 (corresponding to 20 elec-

trons). Therefore the electronic configuration is:
1s2 2s2 2p6 3s2 3p6 4s2
linear Valence shell electron pair repulsion (VSEPR) theory

This is used to predict the shape of molecules. In order to be able to predict the
geometry (or shape) of a molecule several simple steps are required. For exam-
trigonal planar
ple, consider the case of BrF3. In this case we have indicated by underlining that
the central atom is bromine. The first step is to determine the number of valence
electrons for bromine; this is done by establishing the electron configuration
for bromine, i.e. 1s2 2s2 2p6 3s2 3p6 4s2 3d10 4p5. We can determine that the
number of outer electrons is seven (from 4s2 and 4p5). We then determine how
tetrahedral
many atoms are attached to the central bromine atom – the answer is three. By
simple addition we have 7 + 3 = 10 electrons. We know that two electrons are
required per bond; therefore we have enough electrons for five bonds. Then, it
is simply the case to determine a geometry that allows for five bonds to be at
the maximum distance from each other. Commonly, five different arrangements
of atoms are found: linear, trigonal planar, tetrahedral, trigonal bipyramidal
trigonal bipyrimidal
and octahedral. These arrangements are shown in Fig. 55.9. We can therefore
see that the geometry of BrF3 is trigonal bipyramidal. VSEPR also works for
anions and cations using the same procedure. For a cation, a positively charged
species, simply deduct one electron from the total; similarly for an anion, a
negatively charged species, add one electron to the total. In all cases the total
octahedral
number of electrons obtained will be an even number.
Molecular orbital diagrams
Molecular orbital diagrams (see Fig. 55.10) are required in the study of chem-
ical bonding. For a diatomic molecule these consist of two atomic orbitals
Fig. 55.9 Geometries of molecules. (AOs) and two molecular orbitals (MOs). In the case of the two MOs, one is

AO MO AO
O2
O s*6 O
p*2
2p 2p'
p1
s5
s*4
2s 2s'
s3
s*2
1s 1s'
s1
O – electronic configuration: 1s2 2s2 2p4
Fig. 55.10 Molecular orbital diagram for oxygen.
the bonding MO and the other the anti-bonding MO. An asterisk (*) is used
to represent an anti-bonding MO. Bonding MOs (of lower energy) are always
occupied first. Two types of MO are shown, s and p, using oxygen, O2, as an
example. The first task is to determine the electronic configuration for each
oxygen atom, i.e. 1s2 2s2 2p4. An outline of the MO diagram is constructed such
that one AO is located on the left-hand side of the page and the other on the
right-hand side (the use of = indicates that a different AO is represented) with the
MOs positioned in the centre (bonding MOs in the lowest position). You then
have an MO diagram composed of three different energy levels corresponding
to the 1s, 2s and 2p orbitals. Then, by simply adding the correct number of
electrons into the two AOs first, total up the number of electrons. Electrons are
then placed into the MO in the following sequence: lowest available position
first; and then, individual electrons prior to pairing of electrons. The completed
diagram is shown in Fig. 55.10 where all solid lines (in both the AO and MO
positions) represent places where up to two electrons can be paired up (dashed
lines simply indicate association with a particular AO). Arrows ( c T ) indicate
that the electrons are spinning (paired electrons have opposite spins). The num-
bering of bonding and anti-bonding MOs is merely for numerical sequencing
and has no other significance. It is also worth noting that O2 has unpaired
electrons in the anti-bonding MO, i.e. p2*.

Anon. (2015) ChemDraw 15.0. CambridgeSoft Corp., Anon. (2015) ChemSketch 12.0. Available:
Cambridge, USA. Available: http://www.acdlabs.com/download/chemsk
http://www.cambridgesoft.com/ Last accessed 05/03/16.]
Last accessed 05/03/16

Study exercises
55.1 Convert the following abbreviated formulae to (c) (CH3CH2)2C “ O

structural formulae: (d) CH3COCH2COOCH2CH3
CH2 CH2 CH3
(a) BuOH (e) CH3 C CH C
(b) PhCH2Br CH CH2 CH2
(c) PriNH2
55.3 Using VSEPR theory, predict the shapes of the
(d) AcOEt
following molecules:
(e) TsCl
(a) AlCl3
55.2 Convert the following structural formulae to
(b) H 2O
bond-line formulae:
(c) BeF2
(a) CH3CH2CH(OH)CH3 (d) SF6
(b) (CH3)3Cl (e) IF5

56 Chemometrics
Chemometrics has been defined as the chemical discipline that uses mathe-
matical and statistical methods to design or select optimal measurement pro-
cedures and experiments and to provide maximum chemical information by
analysing chemical data. It can assist with (i) the planning of experiments,
and (ii) the manipulation and interpretation of large data sets. Some aspects of
chemometrics can be done using an appropriate spreadsheet but the majority
of applications require the use of dedicated software. The fundamental princi-
ples of most of the processes involved in chemometrics are those of statistics.
You are therefore advised to become familiar with the material in Chapter 53
Table 56.1 Melting points for naphthalene and 54 before proceeding.
When carrying out any experimental work, e.g. an undergraduate practical,
Student m.pt. (°C) you should always read the entire practical script before starting the experimen-
tation. This is important as it allows you to plan each step of the process and
A 79
to organise space and time to perform the experiment. This initial planning is
B 81 further complicated in project work and research projects when, often, there
C 80 is no laboratory script to follow. In these situations, you finally come down to
D 77 planning the initial experiments after background research (e.g. reading the
appropriate scientific literature on the subject area to be investigated), purchas-
E 80
ing/obtaining the appropriate chemicals/reagents, etc. (see also Box 5.1). It is
F 80 at this stage that chemometrics can be of some assistance. Assuming that you
G 79 are able to identify the dependent variables in the experiment, then you can
H 76 apply ‘experimental design’ which allows you to gain the maximum amount
of knowledge about the system you are investigating from a limited number
I 81
of experiments.
J 80 Once the experimental work has been completed you then need to consider
K 79 how to interpret the results, i.e. how to maximise the chemical information
L 78 inherent in the data. Initial attempts are often centred around plotting the data,
M 82
to visualise trends and to allow conclusions to be drawn. The simplest form of
data visualisation is simply to tabulate the results (Chapter 50). As an example,
N 81
if a class of students has determined the melting point of naphthalene, it is a
Mean melting point of naphthalene is 79.5°C. relatively simple matter to tabulate the data (see Table 56.1). One possibility for
the data is then to calculate the mean and standard deviation. Another approach
would be to plot the data as a histogram, as in Fig. 56.1, so we are then able to
5
make a visual interpretation of the quality of this univariate (one-variable) data.
4
However, what if we had more than one variable to consider? In other
frequency
3
words, we have multivariate data. For example, what if we want to identify
2
trends in the properties of a range of organic molecules? The variables we
1
might want to consider could be: melting point, boiling point, Mr, solubility in
0
76 77 78 79 80 81 82 a solvent and vapour pressure. We can, of course, tabulate the data, as before,
melting point (8C) but this does not allow us to consider any trends in the data. To do this we need
to be able to plot the data. However, once we exceed three variables (which we
Fig. 56.1 Histogram: melting point of need to be able to plot in three dimensions) it becomes impossible to produce
naphthalene. a straightforward plot. It is in this context that chemometrics offers a solution,
reducing the dimensionality to a smaller number of dimensions and hence the
ability to display multivariate data. The most important technique in this con-
text is called principal component analysis (PCA).
The following discussion highlights only the basic principles. For more
detailed information you are advised to consult the literature and dedicated
chemometric software packages. It should always be borne in mind, how-
ever, that the choice of which variables to optimise should be selected (i) by

Chemometrics
someone with prior knowledge of the system under investigation, or (ii) after
Useful sources of information Journals:
performing preliminary experiments to determine which are the most impor-
Annual Review of Physical Chemistry tant variables.
Chemical Physics
Chemical Physics Letters Experimental design
International Journal of Quantum There are two main multivariate optimisation strategies: those based on sequen-
Chemistry
tial designs and those based on simultaneous designs.
Journal of Chemical Physics
Journal of Computational Chemistry Sequential design
Journal of Molecular Structure/ Sequential optimisation is based on the one-at-a-time approach. The major
Theochem limitation of this approach is that it assumes that no interaction effects occur
Journal of Physical Chemistry between the variables. Unfortunately this is not always the case. A sequential
Reviews in Computational Chemistry design strategy involves carrying out a few experiments at a time and using the
Theoretica Chimica Acta results of those experiments to determine the next experiment to be done. The
best known of the sequential design approaches is called the simplex method.
A simplex is essentially a geometric figure having a number of vertices equal to
one more than the number of variables. For example, if we have two variables,
the simplex is a triangle, three variables a tetrahedron, and so on.
Let us consider the case of two variables, x1 and x2. An algorithm describes
the initial simplex to be performed (Fig. 56.2). By performing experiments 1–3,
x2
described by the initial simplex, and recording their responses, the next set of
experiments can be described. If we obtain the lowest response for experiment
4 3 2, it can therefore be assumed that a higher response would be obtained in the
opposite direction. By reflecting point 2, we can obtain point 4. By performing
the experiment described by point 4 we obtain its response, thereby perpetu-
1 2 ating the simplex.
Four rules can be described for a simplex design:
x1
1. A new simplex is formed by rejecting the point with the lowest response
and replacing it with its mirror image across the line defined by the two
Fig. 56.2 Simplex optimisation.
remaining points.
2. If the new point in the simplex has the lowest response, return to the
preceding simplex and create the new simplex by using instead the point
with the second-lowest response.
1
3. If a point is retained in three consecutive simplexes, then it can be assumed
6 7 3 4 5 that an optimum has been reached. (Note: it may be that this optimum
is not the true optimum, but that the simplex has been trapped at a false
optimum. In this situation, it is necessary to start the simplex again, or
2
use a modified simplex in which the step size is not fixed but variable, see
Fig. 56.3 Step-size simplex. Fig. 56.3.)
4. If a point is suggested by the simplex algorithm that is beyond the limit
of the variables, i.e. it is beyond the safe working limits of an instrument,
then the point is rejected and an artificially low response is assigned to it,
and the simplex is continued with rules 1–3.
Simultaneous design
In a simultaneous approach the relationship between variables and results is
studied as follows: carry out an appropriate design, apply a mathematical model
to the design, and then apply a response surface method to the data. Appropri-
ate designs might be based on factorial designs (full or fractional) or a central
composite design. Response surface methods frequently rely on visualisation
of the data for interpretation.

Chemometrics
Box 56.1 Example of a two-level factorial design
The recovery of phenols by liquid–liquid extraction, and 1.0 g); and pH (4 and 7). The coded values for the
from 1 litre of river water, and subsequent analysis experiment are shown in Table 56.2. The + 1 values rep-
by high-performance liquid chromatography is to be resent the higher value, e.g. pH7, while - 1 represents
optimised. It has been determined that the following the lower value, e.g. pH4. The number of experiments
are critical to achieving an optimum extraction: the can be reduced if a fractional factorial design is used.
volume of extraction solvent, the mass of salt added For example, in this situation the fractional factorial
and the pH of the water sample. Therefore, a 23 design design would become 23 - 1 = 4 experiments. In this
is required. The experimental levels are: volume of situation the experiments labelled with an asterisk in
extraction solvent (5 and 50 mL); mass of salt added (0.0 Table 56.2 would be done.
Table 56.2 Two-level factorial design Factorial design

In general terms, consider the case of two variables at two levels, e.g. a high
Volume of
Experiment solvent salt pH Result value and a low value. This is termed a two-level design or a (full) 2k factorial
design, where k is the number of variables. Therefore we have 22 = 4 experi-
1 -1 -1 -1 Y1 ments to be done. Often the values of the variables are coded; this is done for
2* +1 -1 -1 Y2 convenience purposes only. In this example, high and low values will be coded
3 +1 +1 -1 Y3 as (+) and (-). Alternatively, it might be the case that the number of variables
is three. In this situation we would have a 23 factorial design, requiring eight
4* -1 +1 -1 Y4
experiments. An example of a two-level factorial design is shown in Box 56.1.
5* -1 -1 +1 Y5 The limitation of the two-level factorial design approach is that no estima-
6 +1 -1 +1 Y6 tion of curvature can be determined. In order to take this into account the use
7 +1 +1 +1 Y7 of designs with at least three levels is required. Three-level designs are there-
fore often known as response surface designs. Probably the most important
8* -1 +1 +1 Y8
design in this context is the central composite design (CCD). Central composite
designs consist of a full (or fractional) factorial design onto which is superim-
posed a star design. The number of experiments to be done (R) can be worked
out as follows:
R = 2k + 2k + n0[56.1]
where k is the number of variables, and n0 is the number of experiments in the

centre of the design.
For a design with three variables we would require [23 + (2 * 3) + 1] = 15
experiments. In order to obtain repeatability information it is necessary to run
an experiment several times. This is done by performing the centre point exper-
iment twice. The total number of experiments would therefore be 16. The list
of experiments is shown in Table 56.3 while Fig. 56.4 shows a diagrammatic
representation of the CCD. The CCD is composed of a 3k factorial design
superimposed with a star design (+ a, -a). In order to minimise systematic
error (bias) it is necessary to randomise the experimental run order. This is
shown in Table 56.4.
Response surface methodology

Response surface methodology allows the relationship between the responses
and variables to be quantified, using a mathematical model, and to be visual-
ised. Thus the equation for a straight-line graph can be written as:
y = mx + c[56.2]

Chemometrics
Table 56.3 Central composite design for three variables
Experiment Variable 1 Variable 2 Variable 3 Result
Factorial design, 23
1 -1 -1 -1 Y1
2 +1 -1 -1 Y2
3 +1 +1 -1 Y3
4 -1 +1 -1 Y4
5 -1 -1 +1 Y5
6 +1 -1 +1 Y6
7 +1 +1 +1 Y7
8 -1 +1 +1 Y8
Star design, 2 k
9 -a 0 0 Y9
10 +a 0 0 Y10
11 0 -a 0 Y11
12 0 +a 0 Y12
13 0 0 -a Y 13
14 0 0 +a Y 14
Centre points
15 0 0 0 Y 15
16 0 0 0 Y 16
Table 56.4 A typical randomised CCD
Original
+
experiment (New)
(from Experiment
Table 66.3) run order Variable 1 Variable 2 Variable 3 Result
3k factorial design star design 8 1 -1 +1 +1 Y1

3 2 +1 +1 -1 Y2
14 3 0 0 +a Y3
6 4 +1 -1 +1 Y4
12 5 0 +a 0 Y5
13 6 0 0 -a Y6
central composite design 4 7 -1 +1 -1 Y7
Fig. 56.4 Central composite design. 1 8 -1 -1 -1 Y8
7 9 1 1 1 Y9
15 10 0 0 0 Y10
10 11 +a 0 0 Y11
16 12 0 0 0 Y12
9 13 -a 0 0 Y13
2 14 +1 -1 -1 Y14
5 15 -1 -1 +1 Y15
11 16 0 -a 0 Y16

Chemometrics
where m is a constant and c is the intercept. This describes the relationship

between a single variable (x) and its response (y). Using the previous exam-
ple, with three variables (x1, x2 and x3) it is possible to extend this mathe-
matical model.
First of all we can consider how each of the variables influences the
response (y) in a linear manner. However, the relationship between y and x1, x2
and x3 may not be linear, so it is necessary to consider the possibility of cur-
vature. This is done in terms of a quadratic variable, i.e. a squared dependence
(x 21, x 22 and x 23). Finally, it is also important to consider the effects of possible
interactions between the variables, x1 S x2 S x3, i.e. x1x2, x1x3 and x2x3. The
1900
overall general equation can therefore be written as:
(mg kg )
-1
1700
1500 Y = b0 + b1x1 + b2x2 + b3x3 + b4x 21 + b5x 22 + b6x 23 + b7x1x2 +
tion
analyte concentra
1300 b8x1x3 + b9x2x3[56.3]

1100
900 where b0 is the intercept parameter and b1- b9 are the regression coefficients
200 for linear, quadratic and interaction effects.
16
160 14 This equation can be analysed using multiple linear regression and
12
120 10 n)
te
m
8 i tested for statistical significance at, for example, the 95% confidence interval
pe
80 ( m
ra
6 e
tim
tu
4 (see p. 499). In addition, the response can be explored by plotting a three-

re
40 2
(8C
)
dimensional graph. Unfortunately, in the above example, three variables are

Fig. 56.5 Example of a response surface. present. This immediately constrains what it is possible to plot on the graph
(one of the axes must be the response). One way to select the two variables
to plot is by considering their statistical significance and then selecting two
variables which are significant at the 95% confidence interval. An alternative
approach might be simply to plot the two variables you might wish to discuss
in your experimental report. A typical response surface is shown in Fig. 56.5.
It can be seen that the ‘time’ variable has a maximum at 8–12 min while the
‘temperature’ variable has a maximum at 160–180°C. Further experiments
might be carried out at these two maxima to determine the repeatability of
the approach. However, it is necessary to plot all variables consecutively to
identify all maxima.
In general, it is important to consider the following issues when carrying
out an experimental design:
●● Carry out repeat measurements for a particular combination of variables, to
determine the repeatability of the approach.
●● To remove systematic error (bias), you should randomise the order in which
experiments are done (p. 38).
●● It is important to eliminate intervariable effects (confounding), i.e. the situ-
ation where one variable is interrelated to another.
●● Often, the large number of experiments to be carried out makes it impossible
to run all of them on the same day. If this happens run your experiments in
discrete groups or ‘blocks’.
Principal component analysis

The use of modern automated instrumentation allows the acquisition of large
amounts of chemical data. As well as simply tabulating the data, other forms of
‘analysis’ are required to interrogate the chemical information contained within
the data. One such approach, enabling the simplification of large data sets by

Chemometrics
X6
X1
regression line X2
pH
PC2
X3
X5
X4
temperature PC1
Fig. 56.6 Determination of principal component 1. Fig. 56.7 Principal component analysis.
reducing the number of independent variables, is principal component analysis

(PCA). The basis of this approach is:
●● To reduce the number of original independent variables into new axes,
so-called principal components, PCs, each of which can be estimated unam-
biguously. The data contained in these new PCs, and which are expressed as
‘scores’, are uncorrelated with each other.
●● To express, in a few PCs, the amount of variation in the data.
●● To have each new PC express a decreasing amount of variation.
An example of the application of PCA is shown in Box 56.2.
Box 56.2 Example of principal component analysis
Consider a chemical reaction where reactants, X and Y, that can be obtained is the scatter of the data on either
produce product Z. The yield of Z is dependent upon the side of the regression line. This is due to random variation
temperature of the reaction and its pH. And suppose that rather than a trend. In this example therefore it is not
the reaction has been carried out by a class of students, possible to extract any further PCs.
providing a large amount of data. By plotting tempera- In eqn [56.4], the coefficients a and b indicate the rela-
ture against pH (Fig. 66.6), we can identify a single new tive importance of the two variables to PC1, and are called
variable, PC1, which obviously contains aspects of pH and factor loadings. In general, therefore, the plotting of fac-
temperature, i.e. tor loadings between any two PCs can provide useful
information as to the relationship of variables to the PCs.
PC1 = a(temperature) + b(pH)[56.4]
Fig. 56.7 shows such a plot. It is seen that variables 1–3
PC1 can then be used to replace the original two varia- contribute strongly (positively) and variable 5 (negatively)
bles. In addition, the direction of PC1 indicates where the to PC1. Variable 6 contributes strongly to PC2, whereas
greatest variation in the data lies. The other information variable 4 contributes significantly to both PCs.

Chemometrics

Beebe, K.R., Pell, R.J. and Seasholz, M.B. (1998) Otto, M. (2007) Chemometrics: Statistics and Computer
Chemometrics: A Practical Guide. John Wiley & Sons Application in Analytical Chemistry, 2nd edn. John
Ltd, Chichester. Wiley & Sons Ltd, Chichester.
Brereton, R.G. (2003) Chemometrics: Data Analysis for Steiner, E. (2008) The Chemistry Maths Book, 2nd edn.
the Laboratory and Chemical Plant. John Wiley & Sons Oxford University Press, Oxford.
Ltd, Chichester. Varmuza, K. and Filzmoser, P. (2009) Introduction to
Brereton, R.G. (2007) Applied Chemometrics for Scientists. Multivariate Statistical Analysis in Chemometrics. CRC
John Wiley & Sons Ltd, Chichester. Press, Boca Raton.
Brereton, R.G. (2009) Chemometrics for Pattern Recogni-
tion. John Wiley & Sons Ltd, Chichester.
Miller, J.N. and Miller, J.C. (2010) Statistics and Chemom-
Harlow.
Study exercises
56.1 Search the scientific literature and find a journal has been used to interrogate data and identify
article in which a factorial design or central com- patterns within the data. Consider how the vis-
posite design has been used to optimise a set of ualisation of data using PCA may be beneficial.
variables. Consider how they selected the bound-
56.3 Consider how to might use either optimisation or
aries of their chosen variables.
pattern recognition techniques in your work, e.g.
56.2 Search the scientific literature and find a journal research project.
article in which a principal component analysis

57 Computational chemistry
A chemist is normally visualised as someone in a white laboratory coat per-

forming an experiment. However, some chemists never actually go into a ‘wet
chemical’ laboratory but utilise computers to perform experiments.
Why do chemistry on computers?
Computational chemistry uses com- ●● Safety: invariably all laboratory experiments carry some risk, associated with
mercially available software to enhance the chemicals or apparatus to be used. It is usual to perform a risk assessment
chemical knowledge. (p. 6) prior to experimental work. Computational chemistry allows the user
to carry out work on ‘dangerous’ chemicals with no risk!
●● Cost: apart from the financial outlay on a computer and associated periph-
erals together with the appropriate software, no further costs are involved,
unlike the experimental laboratory where most chemicals require disposal
after use.
●● Understanding: computational chemistry has the ability to provide a basis
for understanding chemical principles.
Although chemistry has always been primarily about experiments in the labo-
Definitions ratory, it is also true that molecular models and calculations have been essential
parts of the chemist’s toolkit for a very long time. The pioneering computa-
An ab initio method is a quantum-
tional chemists often had little more than home-made molecular models and
mechanical approach which attempts to
their knowledge of maths and chemistry to help them to understand the link
calculate, from first principles, solutions
between basic physical laws and the properties of different molecules. Although
to the Schrödinger wave equation.
there were some spectacular early successes, such as the first insights into the
Molecular mechanics describes a sys- structures of DNA (Watson and Crick, 1953) and proteins (Pauling et al. 1951;
tem in which the energy depends only Ramachandran et al., 1963), computational chemistry as a whole was held back
on the nuclei present. by the sheer complexity of molecules. However, the twentieth century saw
Quantum mechanics describes a molec- two great advances that have transformed the power of computational chem-
ular system in which both electrons and istry to solve real world problems. The first was the development of quantum
nuclei are involved. mechanics, which provided the essential mathematical framework for accurate
A semi-empirical method is a type of prediction of the behaviour of molecules. The second was the development of
quantum-mechanical theory that incor- cheap and fast computers, which have provided the essential number-crunching
porates information derived from exper- power needed for almost all modern computational chemistry.
imental data.
Key Point Today, many chemists do all their research at the
computer, and computational chemistry has become an essen-
tial part of many undergraduate courses. The field has grown
to include a number of overlapping areas. Although the names
of the different branches are often used quite loosely and inter-
changeably we can identify a number of sub-disciplines within
computational chemistry.
Quantum mechanics
Now almost 100 years old, quantum mechanics is a mathematical description
of molecules on their own terms; where the rules of the universe appear shock-
ingly different to those of the everyday world that we are used to. In its early
days, only geniuses like Schrödinger and Dirac could make use of quantum
mechanics, because of its sheer difficulty. However, many years of development
have produced computer programs that do almost all of the hard work, allowing
even novices to ‘borrow’ the knowledge and skill of the writers of the program.

Computational chemistry
There are a number of different types of quantum calculations. Probably the

Density functional theory – This is a
most widely used method at the moment is density functional theory, or DFT.
general purpose quantum method that
Other important flavours of quantum mechanics include Hartree–Fock (HF),
can be used to calculate the properties
which is similar to DFT but a bit more limited in scope, and semi-empirical
of almost any molecule, which provides
methods, which are much faster than DFT and HF, but also much more lim-
the best compromise between speed
ited in their versatility and power. Quite a few quantum chemistry programs
and accuracy for many applications.
are now available; most of them are commercial products, and their power
and ‘user-friendliness’ are quite variable. Most of these programs have been
developed over several decades, and are capable of several different types of
The main commercial quantum chemis- calculation, including DFT, HF and semi-empirical. These have broadly similar
try packages used by researchers include capabilities and performance, although each has its own special strengths. Such
Amsterdam Density Functional (ADF), programs also vary in their degree of ‘user-friendliness’. For example, not all
Gaussian, Spartan and Turbomole. of them have built-in graphical user interfaces (known as a GUI); in the case of
Gaussian, for example, the GUI comes as an optional extra called GaussView.
Several non-commercial graphics programs such as Moldraw and Molden can
be used with one or more of these quantum chemistry programs. Also worthy
of mention here is Hyperchem; although not as powerful as Gaussian or ADF,
Hyperchem is more user-friendly, and as it can perform a range of quantum
calculations it is often preferred for teaching purposes.
Molecular mechanics
The main limitation of quantum mechanics is the size of the calculations. Cal-
culations on typical molecules might take several days to run on a desktop
computer, while larger molecules such as small proteins need a supercomputer.
Therefore, many types of molecule are still beyond the practical reach of rou-
tine quantum calculations, and we need a faster method in order to be able to
model these systems. Molecular mechanics provides a much simpler treatment
of chemical bonds, which is very much faster and so can look at much bigger
molecules. The penalty is that molecular mechanics fails when we are inter-
ested in properties of molecules that depend on their quantum nature; for exam-
ple, chemical reactions. However, molecular mechanics is very useful when
we are interested in the structures of molecules, or how the energy changes
with conformation. This area is often called molecular modelling, and it relies
heavily on graphical interfaces to allow the user to build and edit molecules. As
with quantum mechanics, many different molecular mechanics programs are
available. Some of these are used extensively in industry, particularly by drug
companies, and so tend to be quite expensive (tens of thousands of pounds!).
Examples of these commercial packages include Discovery Studio by Accelrys
and Sybyl by Tripos. Also worth mentioning here is Hyperchem, which again
provides a good introduction to the area at much lower cost. Many academics
use Gromacs, which is a powerful molecular mechanics program available as
open source software, and DeepView, which is a protein homology modelling
program, freely available from the Swiss Institute of Bioinformatics.
What can calculations tell us?

In principle, quantum calculations can tell you almost anything you might wish
to know about a molecule. In practice, of course, some properties are much
easier to calculate than others. The three main properties which are routinely
calculated are:
●● Molecular geometries. This is the most widely calculated property. Geome-
tries can be obtained by both quantum and molecular mechanics. For many

small molecules, the calculated geometry can often be as good as an exper-

imental geometry (e.g. an X-ray crystal structure). However, an important
consideration for all but the simplest of cases is that molecules are not rigid,
but flexible; in particular, some of the bonds can show rotation. In this case,
the molecule may have more than one stable geometry. The problem is then
to find the most realistic model. The structure with the lowest possible energy
is called the global minimum and other stable structures are called local
minima. As modellers, we would like to find the global minimum; but as the
molecule grows in size and complexity, it becomes increasingly more difficult
to find the one global minimum in a large set of possible local minima. It is
worth remembering that molecules in solution do not sit rigidly in their global
minimum structures. Instead, the structure of each individual molecule con-
tinually cycles through all of the allowed conformations, spending most time
around the global minimum. This type of process can be modelled through a
special type of calculation called molecular dynamics (MD). MD simulates
how the structure of a molecule changing over time. Since MD simulations
require a lot of computer time, they are usually run using molecular mechan-
ics rather than quantum mechanics. (See Fig. 57.1 for an illustrative example.)
●● Energies. Here again, both quantum and molecular mechanics can be used to
provide the energies of molecules. However, in thiscasemolecular mechanics
calculations are more limited; one can only compare energies for the same
molecule in different geometries. For example, if you are interested in the
energies of different isomers with the same chemical formula, you have to use
a quantum method. Nevertheless, molecular mechanics energies can be very
useful, for example, in the molecular dynamics simulations mentioned above.
Using quantum calculations, we can also access energy-dependent properties
of molecules, such as redox potentials, pKa values, reaction energies, etc.
●● Spectra. Using reasonably powerful quantum methods, it is possible to cal-
culate many different spectroscopic properties. The most common types of
spectroscopic calculation are UV/visible, IR and NMR; all of these can be
calculated with reasonable accuracy for many types of molecule. Many other
more exotic types of spectrum have been calculated by specialists in the field,
for example, photoelectron, microwave and EPR, to name but a few.
It is worth mentioning that there are many more specialist types of mod-
elling in use for different purposes. To give a few examples, protein homology
modelling is a very useful way of building 3D models for proteins, based on
known experimental structures. Quantitative Structure Activity Relationships
(QSAR) is an important tool in drug development, where the structures of
known drug molecules are used to develop models that allow the prediction of
new biologically active compounds. Chemical engineers routinely use simu-
lation programs to model the behaviour of chemical reactions under different
industrial processing conditions.
The computational laboratory

Like all laboratory classes it is important to go prepared so that you will get the
most out of your time in the laboratory. This might include background reading,
making an outline of the experimental procedure, a sketch (or photocopy) of
the chemical structure of all molecules to be worked on, and a plan of how you
will draw each molecule – obviously the more complex molecules may require
more thought than a simple molecule. Once there, it is important to:

O CH3
O
Aspirin
O
OH
(a) global minimum (b) +3. –kJ mol
(c) +12.5 kJ mol (d) +23.4 kJ mol
Fig. 57.1 Four minimum energy structures for the aspirin molecule. The
geometries were calculated using DFT and the energies are for the gas
phase. Structure (a) has the lowest energy and is therefore referred to
as the global minimum. Structure (b) has a slightly higher energy, while
structures (c) and (d) show progressively higher energies. Structure (b)
corresponds to the experimental solid state structure, as determined by
X-ray crystallography; the small energy difference between structures (a)
and (b) could reflect the differences between the solid and gas phases
as much as inaccuracies in the calculations. A solution of aspirin would
contain all four isomers, in proportions related to their relative energies.
●● Get a comfortable chair – you may be sitting in it for quite a while. Make
yourself feel at ease and relax.
●● Plan your work – a considerable amount of time in the laboratory will be spent
constructing models, setting up calculations and evaluating results. It is there-
fore important to maximise your time on the computer by planning in advance.
●● Make sure that enough computer running time is available – some processes
can be lengthy.
●● Follow all instructions carefully – remember that a computer carries out
your instructions.

●● Examine all results carefully – do not accept everything the computer prints
out/displays. Question the results yourself – do they make sense? If not
recheck your initial data entry.
●● Save all your results for rechecking by yourself at a later date or for assess-
ment by your tutor.
Computer software
A typical software package used to perform computational chemistry should
be able to:
●● Build and display molecules.
●● Optimise the structure of molecules.
●● Investigate the reactivity of molecules.
●● Generate and view orbitals and electronic plots.
●● Evaluate chemical pathways and mechanisms.
●● Study the dynamic behaviour of molecules.
It is inappropriate to describe any particular software system. Nevertheless, all
software is usually accompanied by a user manual or a ‘help’ file to make the
use of commercial software packages user-friendly.
Text references
Watson, J.D. and Crick, F.C.H. (1953) Nature, 171, 737. Ramachandran, G.N., Ramakrishnan, C. and Sasisekharan,
Pauling, L., Corey, R.B. and Branson, H.R. (1951) Proc. V. (1963) J. Mol. Biol., 7, 95.
Nail. Acad. Sci. USA, 31, 205.

Bachrach, S.M. (2014) Computational Organic Chemistry. Leach, A.R. (2001) Molecular Modelling: Principles and
2nd edn. John Wiley & Sons Ltd, Chichester. Applications, 2nd edn. Pearson Education Ltd, Prentice
Filszar, S. (2008) Atomic Charges, Bond Properties and Hall, Harlow.
Molecular Energies. John Wiley & Sons Ltd, Chichester. Leszczynski, J. and Shukla, M. (eds) (2010) Practical
Guha, R. and Bender, A. (eds) (2012) Computational Aspects of Computational Chemistry. Springer Publish-
Approaches in Cheminformatics and Bioinformatics. ing Co., New York.
John Wiley & Sons Ltd, Chichester. Lewars, E. (2010) Computational Chemistry: Introduction
Hayward, D.O. (2002) Quantum Mechanics for Chemists. to the Theory and Applications of Molecular and Quan-
Royal Society of Chemistry, Cambridge. tum Mechanics. Springer, New York.
Heine, T., Joswig, J.-O. and Gelessus, A. (2009) Computa- Puzyn, T., Leszczynski, J. and Cronin, M.T.D. (2009)
tional Chemistry Workbook: Learning Through Exam- Recent Advances in QSAR Studies: Methods and Appli-
ples. John Wiley & Sons Ltd, Chichester. cations. Springer Publishing Co., New York.
Hinchcliffe, A. (2008) Molecular Modelling for Beginners, Young, D.C. (2001) Computational Chemistry: A Practical
2nd edn. John Wiley & Sons Ltd, Chichester. Guide for Applying Techniques to Real World Problems.
John Wiley & Sons Ltd, Chichester.
Jensen, F. (2006) Introduction to Computational Chemistry
2nd edn. John Wiley & Sons Ltd, Chichester.

Study exercise
57.1 Figure 57.2 shows the energy of the chlorine 600

molecule as a function of the Cl9Cl distance. The
500
solid curve and points were calculated by DFT,
and the dotted curve and open points were cal- 400
Energy, kj mol21
culated by molecular mechanics (MM). The DFT
300
curve reaches a minimum at 2.05 Å, and the MM
curve reaches a minimum at 1.98 Å. The experi- 200
mental Cl2 bond length is 1.99 Å.
100
●● Why are the curves very different when the 0

Cl9Cl distance is greater than ∼ 2.1 Å, and which 0 1.5 2 2.5 3 3.5 4
one is correct? CI–CI distance, Å
●● Why does the energy increase very rapidly as Fig. 57.2 Plot of energy against inter-atomic distance
the Cl9Cl distance drops below 2 Å? for the Cl2 molecule.

Study and examination skills
58. The importance of transferable skills 523
59. Managing your time 529
60. Working with others 534
61. Taking notes from lectures and texts 538
62. Learning effectively 544
63. Revision strategies 551
64. Assignments and exams 556
65. Preparing your curriculm vitae 566

58 The importance of transferable skills
This chapter outlines the range of transferable skills and their significance to
Skills terminology – different phrases
chemists. It also indicates where practical skills fit into this scheme. Having a
may be used to describe transferable
good understanding of this topic will help you place your university studies in
skills and associated personal qualities,
a wider context. You will also gain an insight into the qualities that employers
depending on place or context. These
expect you to have developed by the time you graduate. Awareness of these
include: ‘graduate attributes’, ‘personal
matters will help when carrying out personal development planning (PDP) as
transferable skills’ (PTS), ‘key skills’,
part of your studies.
‘core skills’ and ‘competences’.
The range of transferable skills

Table 58.1 provides a comprehensive listing of university-level transferable
skills under six skill categories. There are many possible classifications – and a
different one may be used in your institution or field of study. Note particularly
that ‘study skills’, while important, and rightly emphasised at the start of many
courses, constitute only one area of skills acquired by most university students.
The phrase ‘Practical Skills’ in the title of this book indicates that there is a
Using course materials – study your
course handbook and the schedules
special subset of transferable skills related to work in the laboratory. However,
for each practical session to find out
although this text deals primarily with skills and techniques required for labo-
what skills you are expected to develop
ratory practicals and associated studies, a broader range of material is included.
at each point in the curriculum. Usu-
This is because the skills concerned are important, not only in the chemical
ally the learning outcomes/objectives
sciences but also in the wider world. Examples include time management,
(p. 555) will outline the skills involved.
evaluating information and communicating effectively.
Key Point Chemistry is essentially a practical subject, and

therefore involves highly developed laboratory skills. The impor-
tance that your lecturers place on practical skills will probably
be evident from the large proportion of curriculum time you will
spend on practical work in your course.
The word ‘skill’ implies much more than the robotic learning of, for example,
Example The skills involved in a laboratory routine. Of course, some of the tasks you will be asked to carry
teamwork cannot be developed fully
out in practical classes will be repetitive. Certain techniques require manual
without a deeper understanding of
the interrelationships involved in suc-
dexterity and attention to detail if accuracy and precision are to be attained, and
cessful groups. The context will be the necessary competence often requires practice to make perfect. However, a
different for every group and a flexible deeper understanding of the context of a technique is important if the skill is
approach will always be required, to be appreciated fully and then transferred to a new situation. That is why this
according to the individuals involved text is not simply a ‘recipe book’ of methods and protocols and why it includes
and the nature of the task. background information, tips and worked examples, as well as study exercises
to test your understanding.
Transferability of skills
‘Transferability’ implies that someone with knowledge, understanding or ability
gained in one situation can adapt or extend this for application in a different
context. In some cases, the transfer of a skill is immediately obvious. Take, for
example, the ability to use a spreadsheet to summarise chemical data and create
a graph to illustrate results. Once the key concepts and commands are learned
(Chapter 45), they can be applied to many instances outside chemistry where

The importance of transferable skills
Table 58.1 Transferable skills identified as important in chemistry. The list has been compiled from the UK Quality Assur-
ance Agency for Higher Education Subject Benchmark Statement for Chemistry (2014). Particularly relevant chapters are
shown for the skills covered by this book
Chemistry-related cognitive abilities and skills

In bachelor’s degree with honours programmes, students develop:
●● the ability to demonstrate knowledge and understanding of essential facts, concepts, principles and theories relating to the subject
areas covered in their programme
●● the ability to apply such knowledge and understanding to the solution of qualitative and quantitative problems that are mostly
of a familiar nature
●● the ability to recognise and analyse problems and plan strategies for their solution
●● skills in the generation, evaluation, interpretation and synthesis of chemical information and data
●● skills in the practical application of theory using computational methodology and models
●● skills in communicating scientific material and arguments
●● information technology and data-processing skills, relating to chemical information and data.
Additionally in master’s degree programmes, students develop:
●● the ability to adapt and apply methodology to the solution of unfamiliar problems
●● the ability to assimilate, evaluate and present research results objectively
●● skills required to undertake a research project reporting outcomes that are potentially publishable (in a peer-reviewed publication).
Chemistry-related practical skills
●● an ability to determine hazards associated with carrying out chemical experiments in terms of chemical toxicity, chemical stability
and chemical reactivity and be able to find information to enable effective risk assessments to be carried out
●● skills to handle chemicals safely and carry out experiments and chemical reactions in a safe manner, based on effective risk
assessments
●● skills required for the conduct of documented laboratory procedures involved in synthesis and analysis, in relation to both inor-
ganic and organic systems
●● skills in the monitoring, by observation and measurement, of chemical properties, events or changes, and the systematic and
reliable recording and documentation thereof
●● skills in the operation of standard chemical instrumentation
●● the ability to plan experimental procedures, given well-defined objectives
●● the ability to interpret and explain the limits of accuracy of their own experimental data in terms of significance and underlying
theory.
●● the ability to select appropriate techniques and procedures

●● competence in the planning, design and execution of experiments
●● skills required to work independently and be self-critical in the evaluation of risks, experimental procedures and outcomes
●● the ability to use an understanding of the uncertainty of experimental data to inform the planning of future work.
Professional skills
●● communication skills, covering both written and oral communication with a variety of audiences
●● skills in the employment of common conventions and standards in scientific writing, data presentation, and referencing literature
●● problem-solving skills, relating to qualitative and quantitative information
●● numeracy and mathematical skills, including handling data, algebra, functions, trigonometry, calculus, vectors and complex
numbers, alongside error analysis, order-of-magnitude estimations, systematic use of scientific units and different types of data
presentation
524 Study and examination skills

●● information location and retrieval skills, in relation to primary and secondary information sources, and the ability to assess the
quality of information accessed
●● information technology skills which support the location, management, processing, analysis and presentation of scientific
information
●● basic interpersonal skills, relating to the ability to interact with other people and to engage in teamworking
●● time management and organisational skills, as evidenced by the ability to plan and implement efficient and effective ways of
working
●● skills needed to undertake appropriate further training of a professional nature other relevant professional skills such as business
awareness.
●● problem-solving skills including the demonstration of self-direction, initiative and originality

●● the ability to communicate and interact with professionals from other subjects
●● the ability to make decisions in complex and unpredictable situations
●● the ability to think critically in the context of data analysis and experimental design
●● the ability to work in multi-disciplinary and multi-skilled teams
●● independent learning skills required for continuing professional development.
this type of output is used. This is not only true for similar data sets, but also in
unrelated situations, such as making up a financial balance sheet and creating a
pie chart to show sources of expenditure. Similarly, knowing the requirements
for good graph drawing and tabulation (Chapters 49 and 50), perhaps practised
by hand in earlier work, might help you use spreadsheet commands to make
the output suit your needs.
Other cases may be less clear but equally valid. For example, towards the end
Opportunities to develop and practise of your undergraduate studies you may be involved in designing experiments as
skills in your private or social life – you part of your project work. This task will draw on several skills gained at earlier
could, for example, practise spread- stages in your course, such as preparing solutions (Chapter 8), deciding about
sheet skills by organising personal or numbers of replicates and experimental layout (Chapters 3 and 5) and perhaps
club finances using Microsoft Excel, or carrying out some particular method of observation, measurement or analysis
teamwork skills within any university (Chapters 13–40). How and when might you transfer this complex set of skills? In
clubs or societies you may join. the workplace, it is unlikely that you would be asked to repeat the same process,
but in critically evaluating a problem or in planning a complex project for a new
employer, you will need to use many of the time management, organisational and
analytical skills developed when designing and carrying out experiments. The
Types of PDP portfolio and their benefits – same applies to information retrieval and evaluation and writing essays and dis-
some PDP schemes are centred on aca- sertations, when transferred to the task of analysing or writing a business report.
demic and learning skills, while others
are more focused on career planning. Personal development planning
Some are carried out independently
and others in tandem with a personal
Many universities have schemes for personal development planning (PDP),
tutor or advisory system. Some PDP
which may go under slightly different names such as progress file or profes-
schemes involve creating an online
sional development plan. You will usually be expected to create a portfolio of
portfolio, while others are primarily
evidence on your progress, then reflect on this, and subsequently set yourself
paper-based. Each method has specific
plans for the future, including targets and action points. Analysis of your trans-
goals and advantages, but whichever
ferable skills profile will probably form part of your PDP (Box 58.1). Other
way your scheme operates, maximum
aspects commonly included are:
benefit will be gained from being fully ●● your aspirations, goals, interests and motivations;
involved with the process.
●● your learning style or preference (see p. 550);

Box 58.1 How to carry out a personal skills audit
1. Create a list of appropriate skills. As noted on p. 527, 4. Note actions. This especially applies to skills with
there are many systems for categorising skills. If your low scores in the previous column – and you may
university publishes a specific skill set, e.g. as part of wish to prioritise certain ones. You will need to think
its framework for personal development planning about ways in which you could improve, and this
(PDP) or graduate attributes, then you should use that. may require some research on your part. Is there a
If not, you could adapt the listing in Table 58.1. Your list book you could read? Is there a training workshop
should relate to you personally, your intended career you could attend? Could an extracurricular activity
and any specific skills associated with your intended help you to develop? Should you sign up to speak
qualification. to a skills advisor? It is important that you recognise
that the solution to any deficiencies you perceive lies
2. Lay out your list in table format. You will need to cre-
in your own hands. At university, no one will do the
ate a table using a word processor or spreadsheet
work for you.
program. Your table should have four columns, as
shown in Table 58.2. 5. Add comments and progress notes. Here is where
you can add any comments to amplify or assist with
3. Rate your skills. This may be challenging for many
the action points. The addition of progress notes
students as it is difficult to be objective and tough
implies that you will revisit the list from time to time.
to gauge employer expectations. A confident student
If your university PDP system allows you to add the
may rate a certain skill strongly, whereas a self-critical
list to a portfolio, then do this.
person may consider the same level of skill to be defi-
cient. However, this does not matter too much as you Inevitably, your skills audit will become out of date after a
will effectively be comparing yourself at different period. It will still be useful, however, to look back at it so
stages in your learning, rather than judging yourself that you can see how you have progressed. This will give
against an outside standard. The suggested method a sense of achievement and self-awareness that could
is to use a scale of 1 to 10, with low values indicating be valuable when speaking to academic tutors, careers
that the skill ‘needs lots of development’ and high advisors and potential employers. You may wish to set
values indicating that, for the time being, you feel that up a new list at intervals, perhaps at the start of each
your competence is ‘well above average’. academic year.
Table 58.2 One possible way of creating a personal skills audit. The second row provides guidance about the content
of each column. The third row provides an example of possible content
Comments and notes on

Skill Rating at [date] with notes Proposed actions progress
You should be quite specific. Provide a realistic evaluation This column will note what This column will summarise
It may be a good Idea to sub- of your competence in the you intend to do to try to your progress. You may wish
divide complex skills such as skill at a specific point in time improve the skill. You might to add a revised rating
‘communication’ tick these off as completed
Giving spoken presentations 4/10 [3 March 2011] Wasn’t sat- 1. Read ch 14 in practical skills in Gave secound presentation to tutorial
isfied with presentation to tutorial Chemistry ✓ group; went well, although quite nerv-
group – nervous, a little disorganised ous at start. Slides much better. Make
2. Learn how to use advanced fea-
and ppt too ‘wordy’ sure not to rush the introduction next
tures of PowerPoint ✓
time. 7/10
3. Ask more questions in tutorials ✓
●● your assessment transcript or academic profile information (e.g. record

of grades in your modules);
●● your developing CV (see p. 570).
Taking part in PDP can help you to focus your thoughts about your univer-
sity studies and future career. This is important, as many chemistry degrees do

not lead only to a single, specific occupation. The PDP process will introduce
Definition you to some new terms and will help you to describe your personality and
abilities. This will be useful when constructing your CV and when applying
Employability – ‘the possession by an for jobs.
individual of the qualities and com-
petences required to meet the chang-
ing needs of employers, and thereby Graduate attributes and employability
help realise his or her aspirations and The skills emphasised in chemistry courses (Table 58.1) are sometimes con-
potential in work’. (The Confederation sidered alongside a university-wide framework of graduate attributes that are
of British Industry (CBI) definition of intended to summarise the qualities and skills that an employer might expect
employability) in those with qualifications from your institution. The associated notion of
‘graduateness’ summarises the effect of degree-level experience and learning
on an individual. This in turn is connected with the concept of ‘employability’,
which encompasses those skills and qualities required to gain and maintain
employment. An understanding of these notions is important for every student,
as this not only leads to a better understanding of the value of certain activities
and assessments, but also provides a specialised vocabulary and gives insights
about personal and career development.
At the end of your course, which may seem some time away, you will aim
to get a job and start on your chosen career path. You will need to sell your-
self to your future employer, firstly in your application form and curriculum
vitae (Chapter 65), and perhaps later at interview. Companies rarely employ
bioscience graduates simply because they know how to carry out a particular
lab routine or because they can recall specific facts about their chosen degree
subject. Instead, they will be looking for a range of graduate level skills and
attributes. Typically, for example, they will seek employees who can demon-
strate the ability to work in a team, to speak effectively and write clearly about
their work. All of these skills and attributes can be developed at different stages
during your university studies.
Key Point Factual knowledge is important in degrees with a

strong vocational element, but understanding how to find and
evaluate information is usually rated more highly by employers
than the ability to memorise facts.
Most likely, your future employer(s) will seek someone with an organised
yet flexible mind, capable of demonstrating a logical approach to problems –
someone who has a range of skills and who can transfer these skills to new situ-
ations. Many competing applicants will probably have similar qualifications. If
you want the job, you will have to show that your additional skills and personal
attributes place you above the other candidates.
Text references
McMillan, K.M. and Weyers, J.D.B. (2013) The Study ac.uk/academicinfrastructure/benchmark/statements/
Skills Book, 3rd edn. Pearson Education, Harlow. chemistry.asp
Quality Assurance Agency (2014) Subject Benchmark Last accessed 01/09/15. [Part of the Quality Assurance
Statement – Chemistry. Available: http://www.qaa. Agency Academic Infrastructure.]


Drew, S. and Bingham, R. (2010) The Guide to Race, P. (2007) How to Get a Good Degree: Making the
Learning and Study skills. Gower Publishing Ltd, Most of Your Time at University. 2nd edn. Open Univer-
Aldershot. sity Press, Buckingham.
Study exercises
58.1 Evaluate your skills. Examine the list of skill top- Skills I feel confident about Where demonstrated
ics shown in Table 58.1. Now create a new table
1.
with two columns, like the one shown opposite.
The first half of this table should indicate five 2.
skills you feel confident about and show where 3.
you demonstrated this skill (for example, ‘work-
4.
ing in a team’ and ‘in a first year group pro-
ject in chemistry’). The second half of the table 5.
should show five skills you do not feel confi- Opportunities for
dent about, or that you recognise need devel- Skills that I could develop development
opment (e.g. ‘communicating in verbal form’). 6.
List these and then list ways in which you think
the course material and activities in your current 7.
modules will provide opportunities to develop 8.
these skills, or what activities you might take to 9.
improve them (e.g. ‘forming a study group with
10.
colleagues’).
58.2 Find skills resources. For at least one of the skills 58.3 Analyse your goals and aspirations. Spend a
in the second half of Table 58.1, check your uni- little time thinking what you hope to gain from
versity’s library database to see if there are any university. See if your friends have the same
texts on that subject. Alternatively, carry out a aspirations. Think about and/or discuss how
search for relevant websites (there are many); these goals can be achieved, while keeping the
decide which are useful and bookmark them for necessary balance between university work, paid
future use (Chapter 43). employment and your social life.

59 Managing your time
One of the most important activities that you can do is to organise your personal
Definition and working time effectively. There is a lot to do at university and a common
complaint is that there isn’t enough time to accomplish everything. In fact,
Time management – a system for con- research shows that most people use up a lot of their time without realising it
trolling and using time as efficiently and through ineffective study or activities such as extended coffee breaks. Develop-
as effectively as possible. ing your time management skills will help you achieve more in work, rest and
play, but it is important to remember that putting time management techniques
into practice is an individual matter, requiring a level of self-discipline not
unlike that required for dieting. A new system won’t always work perfectly
straight away, but through time you can develop a system that is effective for
you. An inability to organise your time effectively, of course, results in feelings
of failure, frustration, guilt and being out of control in your life.
Setting your goals

The first step is to identify clearly what you want to achieve, both in work
and in your personal life. We all have a general idea of what we are aiming
for, but to be effective, your goals must be clearly identified and priorities
allocated. Clear, concise objectives can provide you with a framework in
which to make these choices. Try using the ‘SMART’ approach, in which
objectives should be:
●● Specific – clear and unambiguous, including what, when, where, how and
Example The objective ‘to spend why.
an extra hour each week on directed
study in microbiology next term’ fulfils ●● Measurable – having quantified targets and benefits to provide an under-
the SMART criteria, in contrast to a standing of progress.
general intention ‘to study more’.
●● Achievable – being attainable within your resources.
●● Realistic – being within your abilities and expectations.
●● Timed – stating the time period for completion.
Having identified your goals, you can now move on to answer four very
important questions:
1. Where does your time go?
2. Where should your time go?
Advantages of time management – 3. What are your time-wasting activities?
these include: 4. What strategies can help you?
●● a feeling of much greater control
over your activities; Analysing your current activities
●● avoidance of stress;
●● improved productivity – achieve The key to successful development of time management is a realistic knowl-
more in a shorter period; edge of how you currently spend your time. Start by keeping a detailed time log
●● improved performance – work to for a typical week (Fig. 59.1), but you will need to be truthful in this process.
higher standards because you are in Once you have completed the log, consider the following questions:
charge;
●● increase in time available for non- ●● How many hours do I work in total and how many hours do I use for
work matters – work hard, but play relaxation?
hard too.
●● What range of activities do I do?

Managing your time
Activity Notes
Time
slots
7.00–7.15
7.15–7.30
7.30–7.45
7.45–8.00
8.00–8.15
8.15–8.30
8.30–8.45
8.45–9.00
9.00–9.15
Fig. 59.1 Example of how to lay out a time log. Write activities along the
top of the page, and divide the day into 15-min segments as shown. Think
beforehand how you will categorise the different things you do, from the
mundane (laundry, having a shower, drinking coffee, etc.) to the well-
timetabled (tutorial meeting, sports club meeting) and add supplemen-
tary notes if required. At the end of each day, place a dot in the relevant
column for each activity and sum the dots to give a total at the bottom of
the page. You will need to keep a diary like this for at least a week before
you see patterns emerging.
●● How long do I spend on each activity?

Quality in time management – avoid
spending a lot of time doing unproduc- ●● What do I spend most of my time doing?
tive studying, e.g. reading a textbook
●● What do I spend the least amount of my time doing?
without specific objectives for that read-
ing. Make sure you test your recall of ●● Are my allocations of time in proportion to the importance of my
the material, if you are working towards activities?
an examination (p. 548).
●● How much of my time is ineffectively used, e.g. for uncontrolled social-
ising or interruptions?
If you wish, you could use a spreadsheet (Chapter 45) to produce graphical
summaries of time allocations in different categories as an aid to analysis and
management. Divide your time into:
●● Committed time – timetabled activities involving your main objectives/
goals.
●● Maintenance time – time spent supporting your general life activities (shop-
ping, cleaning, laundry, etc.).
●● Discretionary time – time for you to use as you wish, e.g. recreation, sport,
hobbies and socialising.
Avoiding time-wasting activities

Look carefully at those tasks that could be identified as time-wasting activities.
They include gossiping, over-long breaks, uninvited interruptions and even
ineffective study periods. Try to reduce these to a minimum, but do not count

Managing your time
WEEKLY DIARY Week beginning:
Sunday Monday Tuesday Wednesday Thursday Friday Saturday

DATE
7–8 am Breakfast Breakfast Breakfast Breakfast Breakfast

8–9 Preparation Preparation Preparation Preparation Preparation Breakfast
9–10 Breakfast PE112(L) PE112(L) PE112(L) PE112(L) CHEM(P) Travel
10–11 FREE CHEM(L) CHEM(L) CHEM(L) CHEM(L) CHEM(P) WORK
11–12 STUDY STUDY STUDY STUDY STUDY CHEM(P) WORK
12–1 pm STUDY CHEM(L) CHEM(L) CHEM(L) CHEM(L) TUTORIAL WORK
1–2 Lunch Lunch Lunch Lunch Lunch Lunch Lunch
2–3 (VOLLEY- CHEM(P) STUDY SPORT PE112(P) STUDY WORK
3–4 BALL CHEM(P) STUDY (VOLLEY- PE112(P) STUDY WORK
4–5 MATCH) CHEM(P) STUDY BALL PE112(P) SHOPPING WORK
5–6 FREE STUDY STUDY CLUB) STUDY TEA ROTA WORK
6–7 Tea Tea Tea Tea Tea Tea Tea
7–8 FREE* STUDY STUDY FREE* STUDY FREE* FREE
8–9 FREE* STUDY STUDY FREE* STUDY FREE* FREE
9–10 FREE* FREE* STUDY FREE* STUDY FREE* FREE
Study (h) 2 10 11 4 11 6 0
Other (h) 13 5 4 11 4 9 15
Total study time = 44 h
Fig. 59.2 A weekly diary with examples of entries for a first-year science
student with a Saturday job and active membership of a volleyball club.
Note that ‘free time’ can change to ‘study time’ in sessions marked with
an asterisk when assessed work is to be produced or during revision for
exams. Study time (including attendance at lectures, practicals and tutori-
als) thus represents between 42% and 50% of the total time.
on eliminating them entirely. Remember also that some relaxation should be

programmed into your daily schedule.
Being assertive – if friends and col- Organising your tasks

leagues continually interrupt you, find Having analysed your time usage, you can now use this information, together
a way of controlling them before they with your objectives and prioritised goals, to organise your activities, both on a
control you. Indicate clearly on your short-term and a long-term basis. Consider using a diary-based system (such as
door that you do not wish to be dis- those produced by Filofax, TMI and Day-Timer) that will help you plan ahead
turbed and explain why. Otherwise, try and analyse your progress.
to work away from disturbance. Divide your tasks into several categories, such as:
●● Urgent – must be done as a top priority and at short notice (e.g. doctor’s
appointment).
●● Routine – predictable and regular and therefore easily scheduled (e.g.
Use checklists as often as possible –
post your lists in places where they are
attending lectures or playing sport).
easily and frequently visible, such as ●● One-off activities – usually with rather shorter deadlines and which may be of
in front of your desk. Ticking things off high priority (e.g. a tutorial assignment or seeking advice on a specific issue).
as they are completed gives you a feel-
ing of accomplishment and progress,
●● Long-term tasks – sometimes referred to as ‘elephant tasks’ that are too
increasing motivation.
large to ‘eat’ in one go (e.g. learning a language). These are best managed by
scheduling frequent small ‘bites’ to achieve the task over a longer timescale.

Managing your time
You should make a weekly plan (Fig. 59.2) for the routine activities, with
Matching your work to your body’s
gaps for less predictable tasks. This should be supplemented by individual daily
rhythm – everyone has times of day
checklists, preferably written at the end of the previous working day. Such plans
when they feel more alert and able to
and checklists should be flexible, forming the basis for most of your activities
work. Decide when these times are for
except when exceptional circumstances intervene. The planning must be kept
you and programme your work accord-
brief, however, and should be scheduled into your activities. Box 59.1 provides
ingly. Plan relaxation events for periods
tips for effective time management during your studies.
when you tend to be less alert.
Key Point Review each day’s plan at the end of the previ-
ous day, making such modifications as are required by circum-
stances, e.g. adding an uncompleted task from the previous day
or a new and urgent task.
Box 59.1 Tips for effective planning and working
●● Set guidelines and review expectations regularly. ●● Know your own body rhythms: e.g. are you a morning
person or an evening person?
●● Don’t procrastinate: don’t keep putting off doing
things you know are important – they will not go away ●● Learn to recognise the benefits of rest and relaxation
but they will increase to crisis point. at appropriate times.
●● Don’t be a perfectionist – perfection is paralysing. ●● Take short but complete breaks from your tasks –
come back feeling refreshed in mind and body.
●● Learn from past experience – review your manage-
ment system regularly. ●● Work in suitable study areas – keep your own work-
space organised.
●● Don’t set yourself unrealistic goals and objectives –
this will lead to procrastination and feelings of failure. ●● Avoid clutter (physical and mental).
●● Avoid recurring crises – they are telling you that some- ●● Learn to access and use information effectively
thing is not working properly and needs to be changed. (Chapter 42).
●● Learn to concentrate effectively – do not let yourself be ●● Learn to read and write accurately and quickly
distracted by casual interruptions. (Chapters 61 and 68).
●● Learn to say ‘no’ firmly but graciously when appropriate.

Anon. Day-Timer. Available: http://www.daytimer.co.uk Anon. Study Guides and Strategies. Available: http://www.
Last accessed 01/09/15. studygs.net/timman.htm
[Website for products of Day-Timers Europe Ltd, Last accessed 01/09/15.
Fairfax Road, Newton Abbot, TQ12 6UD, UK.] [Website guides to time management and other study
Anon. Filofax. Available: http://www.filofax.co.uk skills for students.]
Last accessed 01/09/15. Evans, C. (2008) Time Management for Dummies, UK edn.
[Website for products of Filofax UK, Broadgate Tower, John Wiley & Sons Ltd, Chichester.
Primrose Street, London, EC2A 2EW, UK.]

Managing your time
Study exercises
59.1 Evaluate your time usage. Compile a spread- leave low priority activities to ‘fill in’ the spare
sheet to keep a record of your daily activities in time that you may identify. This task should be
15-min segments for a week. Analyse this graph- done on a regular (monthly) basis to allow for
ically and identify areas for improvement. changing situations.
59.2 List your short-, medium- and long-term tasks 59.3 Plan an ‘elephant’ task. Spend some time plan-
and allocate priorities. Produce several lists, one ning how to carry out a large or difficult task
for each of the three timescales, and prioritise (learning a language or learning to use a com-
each item. Use this list to plan your time man- plex computer program) by breaking it down into
agement, by scheduling high priority tasks and achievable segments (‘bites’).

60 Working with others
It is highly likely that you will be expected to work with fellow students during
Definitions practicals, tutorials and group study exercises. This might take the form of
sharing tasks or casual collaboration through discussion, or it might be formally
Team – a team is not simply a group of
directed teamwork such as problem-based learning (Box 63.1) or preparing
people with job titles, but a congrega-
a poster (Chapter 66). Interacting with others in this way can be extremely
tion of individuals, each of whom has a
role this a shared task which is under- rewarding and realistically represents the professional world, where teamwork-
stood by other members. Members of ing is common. The advantages of such collaborative work with others include:
a team may seek out certain roles and ●● Synergistic interactions – teamwork often results in better ideas, produced
they may perform most effectively in the by the interchange of views, and better output, due to the complementary
ones that are most natural to them.
skills of team members.
Team role – a tendency to behave, con-
●● Support provided to individuals by other members of the team.
tribute and interrelate with others in a
particular way. ●● Enhanced levels of personal commitment because individuals will not
(both after Belbin, 2010) want to let the team down.
●● Sharing of responsibilities for complex or difficulit tasks.
However, some people approach teamwork with negative feelings. Rea-
sons for this include:
Peer assessment – this term applies
to marking schemes in which all or a ●● Initial reservations about working with strangers – not knowing whether
proportion of the marks for a team- you will be able to form a friendly and productive relationship.
work exercise are allocated by the team ●● Lack of personal commitment to the specified task – perhaps due to unfa-
members themselves. Read the instruc- miliarity or perceived dislike concerning the topic.
tions carefully before embarking on the
exercise, so you know which aspects of ●● Apprehension that others may not do their share of the work – especially
your work your fellow team members if you hope to do well in the course.
will be assessing. When deciding what ●● Lack of previous experience – worries about the kinds of personal interac-
marks to allocate yourself, try to be as tions likely to occur and the team role likely to suit you best.
fair as possible with your marking.
●● Concerns about the outcomes of peer assessment – in particular, whether
others will give you a fair mark for your efforts.
In most cases, these feelings can be transformed through positive par-
ticipation and a common desire to perform well, so that the team as a whole
achieves its target.
Teamwork skills
Some of the key skills you will need to develop to maximise the success of your
teamworking activities include:
Gaining confidence through experience –
the more you take part in teamwork, the ●● Interpersonal skills. These include the way in which you react to new
more you know how teams operate and people, how you listen to and communicate with others, and how you deal
how to make teamwork effective for you. with conflicts and disagreements.
●● Delegation and sharing of tasks. This implies that you are willing to share
effort and responsibility, are willing to trust your fellow team members, and have
mechanisms to deal with unexpected outcomes or even failure to contribute.
●● Effective listening. This involves giving others the opportunity and time to
express their views and to take these seriously, however expressed.
●● Speaking clearly and concisely. Effective communication is a vital part of
teamwork, not only between team members, but also when presenting team
outcomes to others (see Chapter 67).

Working with others
●● Providing constructive criticism. It is all too easy to be negative but only

Web-based resources and support for
constructive criticism will have a positive effect on interactions with others.
brainstorming – websites such as
http://www.belbin.com give further
information and practical advice for The dynamics of teamworking
teamworking. It is important that team activities are properly structured so that each member
knows what is expected of them. Allocation of responsibilities usually requires
the clear identification of a leader. Several studies of groups have identified
different team roles that derive from differences in personality. You should be
aware of such categorisations, both in terms of your own predispositions and
those of your fellow team members, as it will help the group to interact more
productively. Belbin (2010) identified eight such roles, recently extended to
nine, as shown in Table 60.1. Several of the categories shown in Table 60.1 are
suitable for a leader, including ‘coordinator’ and ‘shaper’.
In formal team situations, your course organiser may deal with these
issues; even if they do not, it is important that you are aware of these roles and
their potential impact on the success or failure of teamwork. You should try to
identify your own ‘natural’ role. If asked to form a team, bear these different
Table 60.1 A summary of the team roles described by Belbin (2010). A good team requires members who are able to
undertake appropriate roles at different times. Each role provides important strengths to a team, and its compensatory
weaknesses should be accepted within the group framework.
Personality Typical function in a

Team role characteristics team Strengths Allowable weaknesses
Coordinator Self-confident, calm and Leading: causing others Good at spotting others’ Often less creative or
controlled to work towards shared talents and delegating intellectual than others
goals activities in the group
Shaper Strong need for Leading: generating Providing drive and Can be headstrong,
achievement; outgoing; action within team; realism to group emotional and less
dynamic; highly strung imposing shape and activities patient than others
pattern to work
Innovator Individualistic, Generating action; Creative, innovative and Tendency to work in
serious-minded; often imposing shape and knowledgeable isolation; ideas may not
unorthodox pattern to work activities always be practical
Monitor-evaluator Sober, unemotional and Analysing problems and Shrewd judgement May work slowly;
prudent evaluating ideas not usually a good
motivator
Implementer Well-organised and Doing what needs to be Organising abilities and Lack of flexibility and
self-disciplined, with done common sense tendency to resist new
practical common sense ideas
Teamworker Sociable, mild and Being supportive, Good listener; reliable Not comfortable
sensitive perceptive and and flexible; promotes when leading; may be
diplomatic; keeping the team spirit indecisive
team going
Resource investigator nthusiastic,
Extrovert, e Exploiting opportunities; Quick thinking; good May lose interest
curious and finding resources; exter- at developing others’ rapidly
communicative nal relations ideas
Completer-finisher Introvert and anxious; Ensuring completion of Good focus on fulfilling Obsessive about details;
painstaking, orderly and activity to high standard objectives and goals may wish to do all the
conscientious work to control quality
Specialist Professional, Providing essential skills Commitment and tech- Contribute on a narrow
self-motivated and nical knowledge aspect of project; tend
dedicated to be single-minded

Working with others
roles in mind during your selection of colleagues and your interactions with
them. The ideal team should contain members capable of adopting most of
these roles. However, you should also note the following points:
●● People will probably best fit one of these roles naturally as a function of
their personality and skills.
●● Group members may be suited to more than one role.
●● In some circumstances, team members may be required to adapt and
take a different role from the one that they feel suits them.
Recording group discussions – make
sure you structure meetings (includ- ●● No one role is ‘better’ than any other. For good teamwork, the group
ing writing agendas) and note their should have a balance of personality types present.
outcomes (taking minutes and noting
action points).
●● People may have to adopt multiple roles, especially if the team size is small.
Key Point In formal teamwork situations, be clear as to how

individual contributions are to be identified and recognised.
This might require discussion with the course organiser. Make
sure that recognition, including assessment, is truly reflective of
effort. Failure to ensure that this is the case can lead to disputes
and feelings of unfairness.
Your lab partner(s)

Making the most of lab partnerships –
make sure that you share the different Many laboratory sessions in the life sciences involve working in pairs or small
practical tasks across all members of groups. In some cases, you may work with the same partner(s) for a series of
your lab group, otherwise some will practicals or for a complete module. The relationship you develop as a team is
misss out on the opportunities afforded important to your progress, and can enhance your understanding of the material
by practical classes. This is particularly and the grades you obtain. Tips for building a constructive partnership include:
important in subjects involving a prac-
●● Introduce yourselves at the first session and take a continuing interest in
tical exam at the end of term.
each other’s interests and progress at university.
●● At appropriate points, discuss the practical (both theory and tasks) and
your understanding of what is expected of you.
●● Work jointly to complete the practical effectively, avoiding the situation
where either partner dominates the activities and gains most from the prac-
tical experience.
●● Share tasks according to your strengths, but do this in such a way that one
partner can learn new skills and knowledge from the other.
●● Make sure you ask questions of each other and communicate any doubts
about what you have to do.
●● Discuss other aspects of your course, e.g. by comparing notes from lec-
tures or ideas about in-course assessments.
●● Consider meeting up outside the practical sessions to study, revise and
discuss exams.
Tutorial groups
Tutorials are a group learning environment. Discussions in tutorials can help
you to develop your ideas by observing others’ approaches, views and ideas
and assimilating the facts and concepts that they introduce (see p. 552). This

Working with others
depends on all present participating fully and enthusiastically, while at the

Making the most of tutorials – most
same time respecting the right of thers to make a contribution and express
tutorials require preparation in advance
their opinions.
of each session: this can include pre-
If you feel you need to team up with someone for study, but lack a suitable
tutorial reading(s), literature-based
’study buddy’, then remember that your fellow tutees might be feeling the same.
research and/or the completion of
Tutorial meetings are therefore chances to set up further informal meetings for
specific tutorial ‘questions’ (see also
study or exam revision.
p. 552).
Collaboration for learning

Much collaboration is informal and consists of pairs or groups of individuals
getting together to exchange materials and ideas while studying. It may consist
of a ‘brainstorming’ session for a topic or piece of work, or sharing efforts to
Studying with others – teaming up
research a topic. This has much to commend it and is generally encouraged.
with someone else on your course for
However, it is vital that this collaborative learning is distinguished from the col-
revision (a ‘study buddy’) is a potentially
laborative writing of individually assessed documents: the latter is not usually
valuable activity and may especially suit
acceptable and, in its most extreme form, is plagiarism, usually with a heavy
some types of learners (p. 550). It can
potential punishment in university assessment systems. Make sure you know
help keep your morale high when things
what plagiarism is (p. 386), what unacceptable collaboration is, and how they
get tough. You might consider:
are treated within your institution.
●● sharing notes, textbooks and other
information;
●● going through past papers together, Key Point Collaboration is inappropriate during the final
dissecting the questions and plan-
phase of an assessed piece of work unless you have been
ning answers;
●● talking to each other about a topic
directed to produce a group report. Collaboration is encouraged
(good for aural learners; Box 5.1); during research and learning activities but the final write-up
●● giving tutorials to each other about must normally be your own work. The extreme of producing
parts of the course that have not copycat write-ups is regarded as plagiarism (p. 386) and will be
been fully grasped. punished accordingly.
Text reference Source for further study

Belbin, R.M. (2010) Team Roles at Work, 2nd edn. Belbin, R.M. Belbin® Team Roles. Available: http://www.
Butterworth-Heinemann, Oxford. belbin.com/Last accessed 01/09/15.
Study exercises
60.1 Evaluate your ‘natural’ team role(s). Using for developing aspects where you feel you might
Table 60.1 as a source, decide which team role have done better.
best fits your personality.
60.3 Reflect upon your teamwork abilities. Draw up a
60.2 Keep a journal during a group activity. Record list of your reactions to previous efforts at collabo-
your feelings and observations about experi- ration or teamwork and analyse your strengths and
ences of working with other students. After the weaknesses. How could these interactions have
event, review the journal, then draw up a strategy been improved or supported more effectively?

61 Taking notes from lectures and texts
Note-taking is an essential skill that you will require in many different situa-
Choose note-taking methods appro
tions, such as:
priately – the method you choose to
take notes might depend on the subject; ●● listening to staff in lectures, seminars and practical classes;
the lecturer and their style of delivery;
●● attending meetings and tutorials;
and your own preference.
●● reading texts and research papers;
●● finding information on the web.
Compare lecture notes with a colleague –
looking at different sets of notes for the
KEY POINT Good performance in assessments and exams is
same lecture may reveal interesting dif-
built on effective learning and revision (Chapters 62 and 63).
ferences in approach, depth and detail.
However, both ultimately depend on the quality of your notes.
Adjusting to the styles of your lecturers – Taking notes from lectures

recognise that different approaches Taking legible and meaningful lecture notes is essential if you are to make sense
to lecture delivery demand different of them later, but many students find it difficult when starting their university
approaches to note-taking. For exam- studies. Begin by noting the date, course, topic and lecturer on the first page
ple, if a lecturer seems to tell lots of of each day’s notes. Number every page in case they get mixed up later. The
anecdotes or spend much of the time most popular way of taking notes is to write in a linear sequence down the
on examples during a lecture, do not page, emphasising the underlying structure using headings, as in Fig. 68.1.
switch off – you still need to be listen- However, the ‘pattern’ and ‘Mind Map’ methods (Figs 61.1 and 61.2) have their
ing carefully to recognise the impor- advocates: experiment, to see which you prefer.
tant take-home messages. Similarly, if Whatever technique you use, don’t try to take down all of the lecturer’s
a lecture includes a section consisting words, except when an important definition or example is being given, or when
mainly of images, you should still try to the lecturer has made it clear that he/she is dictating. Listen first, then write.
take notes – names of organisms, loca- Your goal should be to take down the structure and reasoning behind the lec-
tions, key features, even quick sketches. turer’s approach in as few words and phrases as possible. At this stage, follow
These will help prompt your memory the lecturer’s sequence of delivery. Use headings and leave plenty of space,
when revising. Do not be deterred by
lecturers’ idiosyncrasies; in every case
you still need to focus and take useful Columns: types to be covered:
notes (See Box 61.1). GC - DBI, DB5 HPLC
[Isotherinal vs, GC
temperature-programmed]
carrier gas: Prof. Dean

N2 chromatography
helium (intro)
L1. 20th September
Columns:
HPLC - C18, C8(non-polar)
Detectors:
HPLC - UV/fluorescence RPHPLC
GC - FID, FCD, NPD
+ ms for both
mobile phase: (polar)
methanol-water
ACN-water
[Isocratic vs, gradient]
Fig. 61.1 An example of ‘pattern’ notes, an alternative to the more com-

monly used ‘linear’ format. Note the similarity to the ‘spider diagram’
method of brainstorming ideas (Fig. 68.2).

Taking notes from lectures and texts
pens
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Fig. 61.2 Example of the ‘Mind Map’ approach to note-taking and ‘brainstorming’. Start at the centre with the overall topic
title, adding branches and sub-branches for themes and subsidiary topics. ‘Basic’ maps consist of a branched hierarchy
overwritten with key words (e.g. shaded portion, above). Connections should be indicated with arrows; numbering
and abbreviations are encouraged. To aid recall and creativity, Buzan (2009) recommend use of colour, different fonts,
three-dimensional doodles and other forms of emphasis (e.g. non-shaded portion, above).
but don’t worry too much about being tidy – it is more important that you get
Example Commonly used abbrevia- down the appropriate information in a readable form. Use abbreviations to save
tions include:
time. Recognise that you may need to alter your note-taking technique to suit
E there are, there exist(s) different lecturers’ styles.
6 therefore Make sure you note down references to books and take special care to
∵ because ensure accuracy of definitions and numerical examples. If the lecturer repeats
∝ is proportional to or otherwise emphasises a point, highlight (e.g. by underlining) or make a mar-
S leads to, into gin note of this – it could come in useful when revising. If there is something
d comes from, from you don’t understand, ask at the end of the lecture, or make an appointment to
S S involves several processes in a discuss the matter if there isn’t time to deal with it then. Tutorials may provide
sequence
an additional forum for discussing course topics.
1°, 2° primary, secondary (etc.)
≈ , ≅ approximately, roughly equal to Lectures delivered by PowerPoint or similar presentation programs
= , ≠ equals, not equal to Some students make the mistake of thinking that lectures delivered as
K , [ equivalent, not equivalent to computer-based presentations with an accompanying handout or web resource
6, 7 smaller than, bigger than require little or no effort by way of note-taking. While it is true that you may be
W much bigger than
freed from the need to copy out large diagrams and the basic text may provide
[X] concentration of X
structure, you will still need to adapt and add to the lecturer’s points. Much of the
Σ sum
important detail and crucial emphasis will still be delivered verbally. Furthermore,
∆ change
if you simply listen passively to the lecture, or worse, try to work from the handout
f function
alone, it will be far more difficult to understand and remember the content.
# number
If you are not supplied with handouts, you may be given access to the
∞ infinity, infinite
electronic file, so that you can print out the presentation beforehand, perhaps
You should also make up your own in the ‘3 slides per page’ format that allows space for notes alongside each
abbreviations relevant to the context,
slide (Fig. 61.3). Scan through this before the lecture if you can; then, during
e.g. if a lecturer is talking about gas
the presentation, focus on listening to what the lecturer has to say. Note down
chromatography, you could write ‘GC’
instead, etc. any extra details, points of emphasis and examples. After lectures, you could
also add notes from supplementary reading. The text in presentations can be

Aufbau Principle
• For an electron to be in its ground state, i.e. to

have the lowest possible energy, electrons
must occupy the available orbitals in order of
lowest energy.
• An orbital of lowest energy is always ﬁlled
ﬁrst, before orbitals of higher energy.
S P D F
6D
5F
7S 6P
5D
4F
6S 5P
4D
Increasing energy
5S
4P
3D
4S
3P
3S
2P
2S
1S
9
1s • All you need to remember is

that:
2s 2p
3s 3p 3d • s orbitals can have up to 2 e-
4s 4p 4d 4f • p orbitals can have up to 6 e-

5s 5p 5d 5f 5g
• d orbitals can have up to 10 e-
6s 6p 6d 6f 6g 6h
7s …. • f orbitals can have up to 14 e-
10
Fig. 61.3 An example of a printout from PowerPoint in ‘Handouts (three

slides per page)’ format.
converted to word processor format if you have access to the electronic file. In
Sources of abbreviations for note-taking –
PowerPoint, this can be achieved from the Outline option on the View tab, under
general abbreviations can be found
Presentation Views 7 Normal 7 Outline. You can copy and paste text between
at: http://public.oed.com/how-to-use-
programs in the normal fashion, then modify font size and colour as appropriate.
the-oed/abbreviations/
‘Making up’ your notes
As soon as possible after each lecture, work through your notes, tidying them
up and adding detail where necessary. Add emphasis to any headings you have
made, so that the structure is clearer. If you feel it would be more logical for
your purposes, change the order. Compare your notes with material in a text-
book and correct any inconsistencies. Make notes from, or copy, any useful
material you see in textbooks, ready for revision.
Identifying a study goal – reading
around your subject is worthwhile and Taking notes from books and journal papers
rewarding, but if your time is limited, it Scanning
is important to identify a purpose for
your reading and note-making. Ideally,
Good reading skills are vital when taking notes from written sources. When
the goal should relate to specific learn-
consulting a new source for specific information, the first thing you need to do
ing outcomes or assessments.
is to orientate yourself by understanding the text’s scope and structure. This is
often called ‘scanning’.

●● For books, this might involve a quick inspection of the contents section, a
Validating your sources – in some
check on how each chapter is constructed, and noting, for example, whether
cases texts and references may be rec-
the book has a Glossary that might be useful. Once you are familiar with
ommended by staff, but in others you
the structure and layout, then you might either go directly to the appropriate
may need to evaluate the reliability of
chapter or section, or consult the index.
the source yourself – for example, by
considering whether the author(s) has ●● For research publications, the best idea is to read the Abstract before
a lengthy publication record, who he/ consulting specific text, figures or tables, as this should detail the essen-
she works for or, alternatively, whether tial methods and findings. The paper’s layout thereafter will probably
they might have a biased viewpoint that involve the same component sections (IERaD, p. 602), but occasionally in
could skew their analysis. a journal-specific order. Referencing systems may also differ (p. 382), and
you may wish to note this.
At the next stage, when you are scanning through the text to find material
Definitions
relevant to your goal, it is useful to have a good understanding of how most
writing is structured. Both paragraphs and larger pieces of writing generally
Paraphrasing – This is to quote oth-
include an introduction, a main body and a conclusion. The introductory or
ers’ ideas or points by restating them
‘topic’ sentence/paragraph should orientate you to the material that will follow;
in different words. It is different from
the main body expands on the topic sentence, with explanation and examples;
summary in that you may include more
and the final or ‘terminator’ sentence/paragraph provides a conclusion and/or
detail. For example: Wilson and Brand
a link to the next part.
(2010), based on their studies of the
snail population in Arran, take the view
that small sample size may be irrelevant
when variability is low. KEY POINT Working out how a piece of writing is structured
will help you to scan quickly for relevance, digest the content
Quoting –This involves lifting text directly faster and find material of interest as soon as possible.
from your source and demonstrating
this clearly by using quotation marks
(and sometimes italics), if you wish to Depending on your goal, there may be a specific key word or phrase
miss words out, use ellipsis punctuation for which you are searching, or you may wish to read paragraphs on a spe-
marks (. . . ), as shown in this example: cific topic. As well as an index, most chemistry textbooks use systems of
Wilson and Brand (2010) concluded: ‘this headings and subheadings which facilitate such searches. With digital or
indicates that sample size was small in online resources, you can press the Control and F keys together to access a
this study. . . but that this was acceptable ‘find’ dialog box that will speed this process – however, take care to enter
in view of the low variability encoun- correctly spelled and relevant words, using alternatives if an initial search
tered’. Here, the words ‘because of draws a blank.
financial constraints’ have been omitted Finally, you need to absorb the content and decide what the salient points
as irrelevant to the author’s main point. are (Chapter 42). These will obviously depend on the task you are carrying
For longer quotes (say 40 words or more) out. When taking notes, it is vital that you distinguish between your own
create a separate paragraph of text with paraphrasing or summarising of the text and situations where your notes are
a citation (normally at the end). direct quotes, or you may be guilty of plagiarism (p. 386). If transcribing text
Summarising – This is taking the essence word-for-word, always use quotation marks and always note the full citation
from a text and expressing it briefly in details of the source (p. 382) at the same time.
your own words. For example: Other
researchers defend the use of small sam- Skimming
ple sizes (e.g. Wilson and Brand, 2010).
This is a valuable way to gain the maximum amount of information in the
In all cases above, note the citation
minimum amount of time, by reading as little of a text as is required. Essen-
of a relevant reference, in this case
using Harvard style (author names and
tially, the technique (also termed ‘surveying’) requires you to look at the
date, p. 382). Full details of the source
organisation of the text, rather than the detail. In a sense, you are trying to
should be provided in a reference
see the writer’s original plan and the purpose behind each part of the text.
or bibliography section (p. 383.) See
Look through the whole of the piece first, to gain an overview of its scope
also Chapter 42 (especially Box 42.1) in
and structure. Headings provide an obvious clue to structure, if present. Next,
relation to plagiarism.
look for the ‘topic sentence’ in each paragraph (p. 594), which is often the

first. You might then decide that the paragraph contains a definition that is
Printing PowerPoint slides – use the
important to note, or it may contain examples, so may not be worth reading
‘Black and White’ option on the Print
in detail for your purpose.
menu to avoid wasting ink on printing
When you have found relevant material, note-taking fulfils the vital pur-
of coloured backgrounds. If you wish to
pose of helping you understand and remember the information. If you simply
use colour, remember that slides can be
read it, either directly or from a photocopy, you risk accomplishing neither. The
difficult to read if printed in small for-
act of paraphrasing (using different words to give the same meaning) makes
mat. Always print a sample page before
you think about the meaning and forces you to express this for yourself. It is
printing the whole lecture.
an important active learning technique. A popular method of skimming and
note-taking is called tire SQ3R technique (Box. 61.1).
Scanning effectively – you need to stay
focused on your key words, otherwise
you may be distracted by apparently KEY POINT Obtaining information and then understanding it
interesting but peripheral material. are distinct, sequential, parts of the process of learning. As dis-
cussed in Chapter 61 (Table 61.1), you must be able to do more
Spotting sequences – writers often than recall facts to succeed.
number their points (firstly, secondly,
thirdly, etc.) and looking for these words
in the text can help you skim it quickly. Methods for finding and evaluating texts and articles are discussed further in
Chapters 41 and 42.
Making sure you have all the details
of a source – when taking notes from
a text or journal paper: (a) always take
full details (Chapter 41); (b) if copying
word-for-word make sure you indicate
this using quotes and take special care
to ensure you do not alter the original
wording.
Box 61.1 The SQ3R technique for skimming texts
Survey Gain a quick overview of the contents of the Recall Recite to yourself what has been stated
book or chapter, perhaps by rapidly reading every few paragraphs. Write notes of this
the contents page or headings if appropriate, paraphrasing the text rather
than copying it
Question Ask yourself what the material covers
and how precisely it relates to your study Review Think about what you have read and/or
objectives review your notes as a whole. Consider
where it all fits in
Read Now read the text, paying attention to the
ways it addresses your key questions

Reference
Buzan, T and Buzan, B. (2009) The Mind Map Book: [Note that there are several free versions of mind mapping
Unlock Your Creativity, Boost Your Memory, Change software available on the web, e.g. FreeMind – try using a
Your Life. Pearson, Harlow. search engine to find relevant examples and then URLs.]
Study exercises
61.1 Experiment with a new note-taking technique. If notes and handouts, investing if necessary in
you haven’t tried the pattern or mind mapping files and folders. This will be especially valuable
methods (Figs 61.1 and 61.2), carry out a trial to at the start of a revision period.
see how they work for you. Research the meth-
61.3 Try out the SQ3R technique. The next time you
ods first by consulting appropriate books or
need to obtain information from a text, compare
websites.
this method (Box 61.1) with others you may have
61.2 Carry out a ‘spring clean’ of your desk area and adopted in the past. Is it faster, and does it aid
notes. Make a concerted effort to organise your your ability to recall the information?
Answers to those study exercises are available of www.pearsoned.co.uk/practicalskills.

62 Learning effectively
There are many different ways of learning and at university you have the free-
Be adaptable – you should be prepared
dom to choose which approach to study suits you best. You should tackle this
to adjust your study methods according
responsibility with an open mind, and be prepared to consider new options.
to the topic, the learning environment
Understanding how you learn best and how you are expected to think about
(p. 551) and the approach of your lec-
your discipline will help you to improve your approach to study and to under-
turers (p. 552): well as your personal
stand chemistry at a deeper level.
earning preferences.
Key Point At university, you are expected to set your own

agenda for learning. There will be timetabled activities, assess-
ments and exam deadlines, but it is your responsibility to decide
how you will study and learn, how you will manage your time,
and, ultimately, what you will gain from the experience. You
should be willing to challenge yourself academically to discover
your full potential.
Thinking about thinking

Aligning your learning with a specific The thinking processes that students are expected to carry out can be pre-
approach – it is important not to read sented in a sequence, starting with less complex thought processes and end-
too much into any formal ‘diagnosis’ ing with more comples processes, each of which builds on the previous level
of your preferred method of learning. (see Table 62.1). The first two categories in this ladder apply to gaining basic
You need to take a flexible approach; knowledge and understanding, important when you first encounter a topic. Pro-
no matter how you prefer to learn, cesses 3 to 6 are those additionally carried out by high-performing university
many of the outputs assessed at uni- students, with the latter two being especially relevant to final-year students,
versity (for example, exam answers researchers and professionals. Naturally, the tutors assessing you will want to
essays and reports) are in a formal reward the deepest thinking appropriate for your level of study. This is often
written format and you will need to signified by the words used in assessment tasks and marking criteria (column
apply what you have learned to these four, Table 62.1, and p. 555), and while this is not an exact relationship, being
format. Also, different aspects of your more aware of this agenda can help you to gain more from your studies and
study will require different approaches, appreciate what is being demanded of you.
irrespective of personal preferences.
For example, practical skills are best
learned by ‘hands-on’ activities –
Identifying your personal learning preferences
learning by doing. Learning style analysis is a system devised by academics to measure the effec-
tiveness of different student learning processes. Several, but generally similar,
models have been developed to explain and assist in the acquisition of knowl-
edge and ideas by students. The four common areas of generalised knowledge
Understanding your learning preferences acquisition are: listening, as in formal lectures, tutorials, audio-visual systems,
– although the notion that we have peer-interactions and team work; reading, as in formal lectures, tutorials, text
‘hard-wired’ individual learning ‘styles’ books, hand-outs, editing, summarising and rewriting notes; visualising, as in
is strongly contested by many academ- lectures, presentations, video and web-based material, highlighting notes and
ics and researchers, in practice it can relevant stories; doing, as in laboratory work, tutorial workshops, project work
be helpful to reflect on your personal and collaborative exercises.
preferences, and how you can make the The overall conclusion was that most students are multi-modal in that they
most of these to enhance your learning. use at least two of the types of knowledge acquisition and for effective learning

Learning effectively
Table 62.1 A ladder of thinking processes, moving from shallower thought processes (top of table) to
deeper levels of thinking (bottom of table). This table is derived from research by Benjamin Bloom et al.
(1956). When considering the cue words in typical question instructions, bear in mind that the precise
meaning will always depend on the context. For example, while ‘describe’ is often associated with relatively
simple processes of recall, an instruction like ‘describe how the human brain works’ demands higher-level
understanding. Note also that while a ‘cue word’ is often given at the start of a question/instruction, this
is not universally so.
Thinking Processes and Example of typical question Other cue words

description (in approximate structure, with cue word used in question
order of increasing ‘depth’ Example in chemistry highlighted instructions
1. Knowledge (knowing facts). You might know that the Describe the general ●● define
If you know information, you elements are organised in a arrangement of elements in ●● list
can remember or recognise format known as the the Periodic Table. ●● state
it. This does not always Periodic Table. ●● identify
mean you understand it
at a higher level.
2. Comprehension. If you You might know that the ele- Distinguish between ●● contrast
comprehend a fact, you ments in the Periodic Table metals and non-metals ●● compare
understand what it means. are organised into columns, in the Periodic Table. ●● distinguish
known as Groups, and rows, ●● interpret
known as Periods.
3. Application. To apply a fact You might be able to take Illustrate the trend in first ●● calculate
means that you can put it to Period 2, within the Periodic ionisation energy by ●● illustrate
use in a particular context. Table, and look at changes in plotting the values against ●● solve
first ionisation energy as the atomic number for ●● show
element increases in atomic elements in Period 2.
number.
4. Analysis. To analyse You might be able to deter- Consider how the electron ●● compare
information means that you mine why the trend in first configuration changes ●● explain
are able to break it down into ionisation energy does not across Period 2. ●● consider
parts and show how these change in a linear manner. ●● infer
components fit together.
5. Synthesis. To synthesise, you You might be able to Test the hypothesis that ●● design
need to be able to extract determine how ‘s’ and ‘p’ ‘dips’ in first ionisation ●● integrate
relevant facts from a body of orbits fill with electrons energy are due to ●● test
knowledge and use these to across Period 2. electron–electron repulsion ●● create
address an issue in a novel as the orbits are filled.
way or create something
new.
6. Evaluation. If you evaluate You might be able to com- Evaluate how the trend ●● review
information, you arrive at ment on trends in other in first ionisation energy ●● assess
a judgement based on its Periods within the Periodic changes in Period 3. ●● consider
importance relative to the Table. ●● justify
topic being addressed.
it is useful for students to identify their strengths and weaknesses. If necessary

this can be achieved by using on-line questionnaires which can be found readily
by using the common on-line search engines. The relationship to learning style
and the chemistry curriculum is outlined in Box 62.1

Box 62.1 How to diagnose your learning preferences using the VAK scheme
Learning approach Description of learning preference Learning style Examples in Chemistry

Visual To learn through seeing. During lectures, students ●● The ability to comprehend
who are visual learners will synthetic mechanisms.
tend to sit at the front of the ●● To understand analytical
lecture room and they will chemistry instrumentation.
often take detailed notes dur-
ing discussions / lectures.
Audio To learn through listening. During lectures, students ●● Attend lectures / tutorials
who are audio learners will and discuss their ideas.
often ask questions, partici- ●● Actively like to engage in
pate in discussions and listen group work.
to others. ●● Repeat back verbal
instructions to check
understanding.
Kinesthetic To learn through moving, touching Kinesthetic learners are very ●● Are enthusiastic to get
or doing. hands-on and may not have going in practical work.
a long attention span for
detailed explanations.
Every person’s learning style is either or a combination of auditory,

visual, or kinesthetic (tactile) in terms of the way he or she learns best. No
particular style is better than the others; it is all about what works best for
the individual.
Key Point Having a particular learning preference or style

does not mean that you are automatically skilled in using meth-
ods generally suited to that type of learner. You must work at
developing your ability to take in information, study and cope
with assessment.
Be active in your learning – a common

theme in all recommended approaches Learning effectively in different settings
to effective learning is activity on the How you learn best depends on your personal characteristics, as noted above,
learner’s part. Thus, note-making dur- but the approaches you use must also suit the different modes of teaching you
ing lectures requires a greater level of encounter.
engagement with the material than the
more passive action of simply listen-
ing; discussing concepts with others
Lectures
involves a deeper degree of thinking These are designed to impart knowledge and understanding efficiently on the
than solo thought processes when part of the staff, who can teach large classes using this method. However, in
reading text (see also Box 63.3). some instances they can simply be note-taking exercises for you as a student.
This may tempt you to approach such teaching in a ‘passive’ frame of mind, but

this is almost certainly not the best way to learn. To get the most from lectures,
Definition you need to:
Take-home message(s) – the main ●● Prepare beforehand by finding out what the topic will be, what the learning
point(s) that a presenter wishes you to outcomes are, and doing some preliminary reading.
remember from what they have said
(perhaps a distillation of important facts ●● Arrive in good time, sitting where you can see and hear the lecturer, with
or a set of conclusions). If a lecture or the right materials for taking notes.
seminar has been particularly detailed
or complex, noting these messages will ●● Listen attentively, particularly when the lecturer is stating what he or she
help you to make sense of it at a later plans to cover; when they indicate that you are expected to write things
time – they may well prove to be useful down word-for-word; when a definition is being presented; when key facts
for assignments and/or exams are stated (often emphasised by repetition); and especially at the end, when
the take-home messages are usually covered.
●● Make useful notes – these will not be a transcript of the lecturer’s words,
but rather a personal digest of the key points that have been made (see
Chapter 61). Even if the material is already provided as a PowerPoint
Lay the foundations for learning – or Word file (or equivalent), your aim is to add value by explaining and
unless you are gifted with photographic interpreting for yourself, and thereby to lay the foundations for memo-
recall, it is not easy to commit informa- rising the content.
tion to your memory unless you have
a conceptual framework of understand- ●● Ask questions if you do not understand something. This might not be at
ing in which to ‘place’ it. This ‘bigger the point of delivery, because things may become clearer when you go over
picture’ comes from reading, listening your notes, read a textbook or talk about the subject with a fellow student.
and thinking at an early stage in the However, it should be as soon as possible after those options are exhausted.
process. There will be opportunities to enquire at tutorials, via email or on-line dis-
cussion boards or at later lectures or meetings.
●● Above all, do attend lectures, even if you feel their value may be limited
in some cases – do not underestimate the way that looking and listening
contribute to understanding and memorisation. Also, ‘cold text’ on the page
rarely has the additional context that a lecture can provide.
Box 62.2 provides some tips for dealing with some of the different teaching
styles of lecturers.
Practicals (lab classes)
In chemistry, practical work is essential, so that your understanding is grounded
Making the most of lab demonstrators – in real examples of chemicals, processes and equipment. Many practicals fol-
these staff or postgraduate students are low the ‘scientific method’ (Chapter 70) and illustrate how to design and carry
paid to help you, so ensure you get good out observations and experiments. They may help you to develop manipulative
value from them by listening to their skills, as well as those associated with data recording, analysis and reporting.
advice and asking them questions. How- To maximise learning from practicals, you should:
ever, do not expect them to hand you
answers on a plate – they will probably ●● prepare as well as you can by reading through the schedule in advance;
be under instruction to make you think ●● make sure that you have all equipment and protective clothing needed;
for yourself.
●● arrive at the lab or assembly point in good time;
●● understand and observe safe practice (Chapter 2);
●● follow the advised procedures;

Box 62.2 How to accommmodate different lectures teaching styles
One challenging aspect of lectures is that no two staff ●● It may be difficult to stay attentive if the lecturer’s
members deliver them in the same way. This means that delivery is quiet, monotonous or mumbled (or all
you will need to adjust your listening and notemaking three). Here, you will need to ensure that you sit where
approaches to suit. For example: you can hear, and listen very carefully for key words
and phrases, rather than changes in intonation, that
●● It may be hard to extract facts and meaning from a lec-
might signal importance or change of emphasis. Do
turer who tells meandering stories or keeps the class
not be afraid to ask the lecturer to speak up if you can-
amused with anecdotes. You must not lose focus when
not hear.
listening and need to be alert when the key points are
mentioned, or use the textbook to fill in the missing ●● You may find it difficult to draw out general princi-
material when making up your notes later on. ples if the lecturer provides great detail about his or
her life in science or current research work. Moreo-
●● You may be distracted if the lecturer uses technology
ver, the latter may be hard to follow if complex jar-
in a way that makes you think about the medium rather
gon and undefined acronyms are used frequently. The
than the message. By all means enjoy the videos,
remedy here may be to use the textbook before and
images and electronic wizardry, but do remember to
after the delivery to create an overarching framework
listen out for the underlying messages and focus your
of understanding.
notes on these.
Getting the most from tutorials – this ●● ask questions when you do not understand;
requires preparation beforehand, for ●● manage your time during practicals and use time when waiting for some-
example, by: thing to happen to start your write-up;
●● carrying out the recommended read-
●● take relevant notes;
ing or tackling the set problems;
●● submitting expected work in good ●● draw up informal figures and tables as you go along;
time;
●● thinking about the issues involved in ●● complete your write-up as soon as possible after the session.
the subject matter; These suggestions are discussed in greater detail in later chapters.
●● preparing any questions you might
have about the topic you have been
asked to study; Tutorials
●● reflecting on the potential relevance These may be designed to fulfil a number of purposes, including:
of the tutorial to your programme of
study. ●● providing guidance, as part of an advising system;
●● reviewing and testing your understanding of material covered elsewhere;
Taking account of tutorial assessment – ●● introducing new material that is better taught in small groups;
the tasks prepared for tutorials may ●● developing and assessing essay-writing and numerical skills through
be marked, and in some cases your appropriate tasks;
participation will be graded. Consult
the course handbook to find out what ●● promoting interactions among students and with the tutor.
proportion of the total mark derives The face-to-face discussions that can occur during tutorials are rarely possible
from tutorials and what the marking in larger classes and they add value by allowing views and understanding to
criteria are. Ensure that you maximise develop through an exchange of opinion and knowledge. Think of your tutor
the marks obtained from this source, as as a potential ally. He or she may be able to explain things differently from
they may be relatively easy to achieve if the lecturer, for example, and thereby help you understand any concepts than
you put in the necessary work. you find difficult. Never feel embarrassed about asking questions – the tutor is
there to provide answers and the chances are that, if you find a subject tricky,

so will the rest of the group. Your tutor may also have or assume respon-
sibilities beyond teaching, and be able to help you with subject choices or
non-academic issues.
e-Learning and distance learning

These modes of learning can be a mixed blessing. On the one hand, they allow
Alternative modes of distance learning – easy access to learning materials at a time of your choosing, while on the other,
the discussion opposite assumes that they assume a personal discipline in your approach to study that may be hard to
e-learning facilities are being used but maintain. Effective learning in this context requires the following:
in some courses the materials are pri-
marily paper-based. If this is the case,
●● making regular and frequent visits to on-line modules and portals;
then finding the best way to learn ●● allocating time to study;
may be difficult. You will need to find a
method of making the material ‘come
●● paying special attention to announcements, message pages or email
alive’, perhaps by purchasing or bor-
systems;
rowing well-illustrated texts or video ●● participating in on-line discussions;
materials (such as BBC documentaries).
●● organising materials provided in digital form – such as lists of ‘live’ links
to website addresses;
●● reading the advised materials and more, whether online, or as texts;
●● making notes as appropriate;
●● using active learning techniques (Chapter 63), rather than passively read-
ing the materials provided;
●● observing staff-set milestones in study, assessment and submission of work;
●● creating personal milestones in study, self-assessment and preparation
of work.
The only person who can judge the effectiveness of your learning at university
is you. Only you will know how much effort you put in to your studies and
what you are expecting, in terms of a mark or grade; only you will know how
comfortable you feel with a particular approach; and only you can respond to
the feedback you have been given, in terms of how you will learn for future
assessments. Be prepared to change your learning methods if you find that
they are unsuccessful or difficult to apply in particular circumstances.
Text references
Anderson, L.W., Krathwohl, D.R. and Bloom, B.S. (2001) Bloom, B., Englehart, M., Furst, E., Hill, W and
A Taxonomy for Learning, Teaching and Assessing: A Krathwohl, D. (1956) Taxonomy of Educational Objec-
Revision of Bloom’s Taxonomy of Education objectives. tives: The Classification of Educational Goals. Hand-
Allyn & Bacon, London book I: Cognitive Domain. Longmans, Green, New York
and Toronto.

Study exercises
62.1 Review your note-taking methods. How well 62.2 Think about your thinking. Read through
suited are they to your needs? How well suited Table 62.1 and consider different thinking pro-
are they to the lecture styles of the academic cesses in relation to: (a) feedback you have
staff? Have you captured the important points, received from a previous assignment, or (b) your
or are you merely transcribing exactly what the next assignment and your intended approach
lecturer says? Where lecturers use PowerPoint to it. Does this reveal that your marks could be
slides, are you listening for, and capturing in suffering because you are not thinking deeply
note form, the additional spoken points and enough or are not capturing your thoughts
examples that the lecturer is adding during the clearly enough in your writing?
live presentation?
Answers to these study exercises are available at www,pearsoned.co.uk/practicalskills.

63 Revision strategies
Where your studies are assessed through an exam, you will need to prepare
Definitions for this by revising and, because you may have a number of exams in a short
period, you will need to be well organised. You will need to align your study
Formative assessments – these may be
effort with the expectations of your lecturers, as explained in the course mate-
mid-term or mid-semester tests and are
rials and through the feedback you have been given on previous assignments
often in the same format as later exams.
They are intended to give you feedback and exams. Adopting active methods of revision that are suited to your learning
on your performance. style (Chapter 62) can also make a significant difference to your performance.
Learning outcomes/objectives –
statements of the knowledge, under- The role of assessment and feedback in your learning
standing or skills that a learner will be
able to demonstrate on successful com- Your starting point when preparing for assessment should always be the learn-
pletion of a module, topic or learning ing outcomes or objectives for each module, topic or learning activity. You
activity. will usually find them in your module handbook. They state in clear terms
Summative assessments – these include what your tutors expect you to be able to accomplish after participating in
end-of-year or end-of-module exams. each element and reading around the topic. Also of value will be marking/
They inform others about the standard assessment criteria or grade descriptors, which state in general terms what level
of your work. ln continuous or ‘in-course’ of attainment is required for your work to reach specific grades. These are more
assessment, the summative elements likely to be defined at faculty/college/school/department level and consequently
are spread out over the course. Some- published in appropriate handbooks or websites. Reading learning outcomes
times these assessments may involve a and grade descriptors will give you a good idea of what to expect and the level
formative aspect, if feedback is given. of performance required to reach your personal goals. Relate them to both
the material covered (e.g. in lectures and practicals, or online) and past exam
papers. Doing this as you study and revise will indicate whether further reading
and independent studying is required, and of what type. You will also have a
much clearer picture of how you are likely to be assessed.
Key Point Use the learning outcomes for your course (nor-
mally published in the handbook) as a fundamental part of your
revision planning. These indicate what you will be expected to
Example A set of learning objectives
taken from an introductory lecture on
be able to do after taking part in the course, so exam questions
inorganic chemistry. are often closely based on them. Check this by reference to past
papers.
□□ Define the following terms:
✓✓ Periodic Table
✓✓ period
✓✓ group
There are essentially two types of assessment – formative and summative,
✓✓ electronic configuration although the distinction may not always be clear-cut (see margin). The first way
✓✓ Aufbau Principle you can learn from formative assessment is to consider the grade you obtained
✓✓ Hund’s Rules in relation to the work you put in. If this is a disappointment to you, then there
□□ Identify selected groups of elements must be a mismatch between your understanding of the topic and the marking
in the Periodic Table, e.g. noble scheme with that of the marker, or a problem in the writing or presentation of
gases, halogens, alkali metals, alka- your assignment. If you do not understand the reason for your grade, contact
line earth metals. your tutor.
□□ Determine the electronic configura- The second way to learn from formative assessment is through the writ-
tion of elements B to Ne. ten feedback and notes on your work. These comments may be cryptic, or
□□ Using the mnemonic for the Aufbau scribbled hastily, so if you do not understand or cannot read them, ask the
Principle, write down the electronic
tutor who marked the work. Most tutors will be pleased to explain how you
configuration for the following ele-
could have improved your mark. If you find that the same comments appear
ments: Pb, Mn, Hg, Sn, Cs, Ti, La, Au,
Pt and Bi. frequently, it may be a good idea to seek help from your university’s academic
support unit. Take along examples of your work and representative feedback

Revision strategies
comments so they can give you the best possible advice. Another suggestion
Using tutors’ feedback – it is always
is to ask to see the work of another student who obtained a good mark, and
worth reading any comments on your
compare it with your own. This will help you judge the standard you should
work as soon as it is returned. If you
be aiming towards.
do not understand the comments, or
are unsure about why you might have
lost marks in an assignment, ask for an Preparing for revision and examinations
explanation while the topic is fresh in
your mind. Before you start revising, find out as much as you can about each exam,
including:
●● its format and duration;
●● the date and location;
●● the types of questions;
●● whether any questions/sections are compulsory;
●● whether the questions are internally or externally set or assessed;
●● whether the exam is ‘open book’, and if so, which texts or notes are
allowed.
Your course tutor is likely to give you details of exam structure and timing
Time management when revising –
well beforehand, so that you can plan your revision; the course handbook and
this is a vital to success and is best
past papers (if available) can provide further useful details (Box 63.2). Always
achieved by creating a revision timeta-
check that the nature of the exam has not changed before you make plans based
ble (Box 63.2)
on past papers.
Filing lecture notes – make sure your

Organising and using lecture notes, assignments
notes are kept neatly and in sequence and practical reports
by using a ring binder system. File the Given their importance as a source of material for revision, you should have
notes in lecture or practical sequence, sorted out any deficiencies or omissions in your lecture notes and practical
adding any supplementary notes or reports at an early stage. For example, you may have missed a lecture or prac-
photocopies alongside. tical due to illness, etc., but the exam is likely to assume attendance throughout
the year. Ask other students for copies of notes of any sessions missed.
Your practical reports and any assignment work will contain specific com-
ments from the teaching staff, indicating where marks were lost, corrections,
mistakes, inadequacies, and so on. Most lecturers are quite happy to discuss
such details with students on a one-to-one basis and this information may pro-
vide you with ‘clues’ to the expectations of individual lecturers that may be
useful in exams set by the same members of staff. However, you should never
‘fish’ for specific information on possible exam questions, as this will not be
well received.
Revision
Recognise when your concentration, Begin early, to avoid last-minute panic. Start in earnest several weeks before-
wavering – take a short break from hand, and plan your work carefully.
revision when this happens and return
●● Prepare a revision timetable – an ‘action plan’ that gives details of specific
to work refreshed and ready to learn.
topics to be covered (Box 63.2). Find out at an early stage when (and where)
Remember that 20 min is often quoted
your examinations are to be held, and plan your revision around this. Try to
as a typical limit to a spell of full con-
centration effort.
keep to your timetable. Time management during this period is as important
as keeping to time during the exam itself.

Revision strategies
Box 63.1 How to use past exam papers in your revision
Past exam papers are a valuable resource for targeting synthesis based on other knowledge? Can you
your revision. identify different styles for different lecturers? Con-
sider how the answers to these questions might
1. Find out where the past (or sample) exam papers are
affect your revision effort and exam strategy.
kept. Copies may be lodged in your department or the
library; or they may be accessible online. 5. Practise answering questions. Perhaps with friends,
set up your own mock exam when you have done
2. Locate and copy relevant papers for your module(s).
a fair amount of revision, but not too close to the
Check with your tutor or course handbook that the
exams. Use a relevant past exam paper; and try not
style of paper will not change for the next set of
to study it beforehand. You need not attempt all of the
examinations.
paper at one sitting. You will need a quiet room in a
3. Analyse the design of the exam paper. Taking into place where you will not be interrupted (e.g. a library).
account the length in weeks of your module, and Keep close track of time during the mock exam and try
the different lecturers and/or topics for those weeks, to do each question in the length of time you would
note any patterns that emerge. For example, can you normally assign to it (see p. 598) – this gives you a feel
translate weeks of lectures/practicals into numbers for the speed of thought and writing required and the
of questions or sections of the paper? Consider how scope of answer possible. Mark each other’s papers
this might affect your revision plans and exam tac- and discuss how each of you interpreted the question
tics, taking into account: (a) any choices or restric- and laid out your answers and your individual mark-
tions offered in the paper; and (b) the different types ing schemes.
of questions asked (i.e. multiple choice, short-answer
6. Practise writing answer plans and starting answers.
or essay).
This can save time compared with the ‘mock exam’
4. Examine carefully the style of questions. Can you approach. Practise in starting answers can help you
identify the expectations of your lecturers? Can get over stalling at the start and wasting valuable
you relate the questions to the learning objectives? time. Writing essay plans gets you used to organising
How much extra reading do they seem to expect? your thoughts quickly and putting your thoughts into
Are the questions fact-based? Do they require a a logical sequence.
Box 63.2 How to prepare and use a revision timetable
1. Make up a grid showing the number of days until your according to difficulty (with the hardest getting the
exams are finished. Divide each day into several sec- most time), or within subjects, according to topics.
tions. If you like revising in large blocks of time, use Make sure there is an adequate balance of time
a.m., p.m. and evening slots, but if you prefer shorter among topics and especially that you do not avoid
periods, divide each of these in two, or use hourly working on the subject(s) you find least interesting
divisions (see also the table in study exercise 63.2). or most difficult.
2. Write in your non-revision commitments, including 4. Allocate the work to the different slots available on
any time off you plan to allocate and physical activity your timetable. You should work backwards from the
at frequent intervals. Try to have about one-third or a exams, making sure that you cover every exam topic
quarter of the time off in any one day. Plan this in rela- adequately in the period just before each exam. You
tion to your best times for useful work – for example, may wish to colour-code the subjects.
some people work best in the mornings, while others
5. As you revise, mark off the slots completed – this has
prefer evenings. If you wish, use a system where your
a positive psychological effect and will boost your
relaxation time is a bonus to be worked for; this may
self-confidence.
help you motivate yourself.
6. After the exams, revisit your timetable and decide
3. Decide on how you wish to subdivide your subjects
whether you would do anything differently next time.
for revision purposes. This might be among subjects,

Revision strategies
●● Study the learning outcomes for each topic (usually published in the
Question-spotting – avoid taking this
course handbook) to get an idea of what lecturers expect from you.
approach in an attempt to reduce the
amount of time you spend revising. ●● Use past papers as a guide to the form of the exam and the type of question
Lecturers are aware that this strategy likely to be asked (Box 63.2).
may be taken and try to ask questions
●● Remember to have several short (five-minute) breaks during each hour
in an unpredictable manner. You may
of revision and a longer break every few hours. In any day, try to work for
find that you are unable to answer on
a maximum of three-quarters of the time.
unexpected topics that you failed to
revise. Moreover, if you have a precon- ●● Include recreation within your schedule: there is little point in tiring your-
ceived idea about what will be asked, self with too much revision, as this is unlikely to be profitable.
you may also fail to grasp the nuances
●● Make your revision as active and interesting as possible (see Box 63.3):
of the exact question set, and provide a
the least productive approach is simply to read and reread your notes.
response lacking in relevance.
●● Ease back on the revision near the exam: plan your revision to avoid
last-minute cramming and overload fatigue.
Box 63.3 How to revise actively
The following techniques may prove useful in devising ●● Draw diagrams from memory: make sure you can
an active revision strategy. label them fully.
●● ‘Distil’ your lecture notes to show the main headings ●● Try recitation as an alternative to written recall. Talk
and examples. Prepare revision sheets with details for about your topic to another person, preferably some-
a particular topic on a single sheet of paper, arranged one in your class. Talk to yourself (in private) if nec-
as a numbered checklist. Wall posters are another use- essary. Explaining something out loud is an excellent
ful revision aid. test of your understanding.
●● Aid recall through effective note-taking: for example, ●● Associate facts with images or journeys if you find this
the mind map technique (p. 543), when used to organ- method works.
ise ideas, is claimed to enhance recall for some by con- ●● Use a wide variety of approaches to avoid boredom
necting the material to visual images or linking it to the during revision (e.g. record information on audio tape,
physical senses. use cartoons, or any other method, as long as it is not
●● Carry out revision checks: it is important to test your- just reading).
self frequently to ensure that you have retained the ●● Form a revision group to share ideas and discuss top-
information you are revising. Assess what you have ics with other students.
learned by taking a blank sheet of paper and writing
down all you know on a selected topic. Check your full ●● Prepare answers to past papers, e.g. write essays or, if
notes to see if you missed anything out. If you did, go time is limited, write essay plans (see Box 63.1).
back immediately to a fresh blank sheet and redo the ●● If your subject involves numerical calculations, work
example. Repeat, as required. through representative problems.
●● Make up lists of key facts and definitions: these can ●● Make up your own questions: the act of putting your-
be a useful starting point for many exam answers. Test self in the examiner’s mindset by inventing questions
yourself repeatedly on these lists, or get a friend to do can help revision. However, you should not rely on
this. Try to remember how many points you need to ‘question-spotting’: this is a risky practice!
know in each case – this will help you recall them all
during the exam. ●● Write key points on Post-it® notes or similar: arrange
and rearrange these in clusters or as lists around your
●● Use mnemonics and acronyms to commit specific study space or room.
factual information to memory. Sometimes, the dafter
they are, the better they seem to work.

Revision strategies
The evening before your exam should be spent in consolidating your mate-
Final preparations – try to get a good
rial, and checking through summary lists and plans. Avoid introducing new
night’s sleep before an exam. Last-
material at this late stage: your aim should be to boost your confidence, putting
minute cramming will be counterproduc-
yourself in the right frame of mind for the exam itself.
tive if you are too tired during the exam.
Key Point When considering assessment questions, look

carefully at words used in the instructions. These cues can
help you identify what depth is expected in your answer (see
Table 62.1). Take special care in multi-part questions, because
the first part may require lower-level thinking, while in later parts
marks may be awarded for evidence of deeper thinking.

Burns, R. (1997) The Student’s Guide to Passing Exams. [Many universities host study skills websites; these can be
Kogan Page, London. found using ‘study skills’, ‘revision’ or ‘exams’ as key-
Hamilton, D. (2003) Passing Exams: A Guide for Maxi- words in a search engine.]
mum Success and Minimum Stress. Cengage Learning, Sheratt, P. (2013) Passing Exams for Dummies., 2nd edn.
Boston. Wiley-Blackwell, London.
McMillan, K.M. and Weyers, J.D.B. (2011) How to Succeed in O’Brien, D. (2007). How to pass Exams: Accelerate
Exams and Assessments, 2nd edn. Prentice Hall, London. your Learning Memorise Key Facts-Revise Effec-
McMillan, K.M. and Weyers, J.D.B. (2012) The Study tively, 2nd Revised edn., Watkins Publishing Ltd,
Skills Book, 3rd edn. Pearson Education, Harlow. London.
Study exercises
63.1 Make use of past exam papers. Use the tech- wish to use or adapt the arrangement below,
niques discussed in Box 63.1 to improve your either on paper or within a spreadsheet.
revision strategy: assess their effectiveness in a
63.3 Try out a new active revision technique. Try
particular exam, or series of exams.
any or all of the methods mentioned in Box 63.3
63.2 Draw up a revision timetable. Use the tech- when revising. Compare notes with a colleague –
niques discussed in Box 63.2 to create a revision which seems to be the most successful technique
timetable for your forthcoming exams. You may for you and for the topic you are revising?
A revison timetable planner.
Date Morning Afternoon Evening

Lunch Tea/Dinner
Session 1 Session 2 Session 1 Session 2 Session 1 Session 2

64 Assignments and exams
Many universities operate a modular system for their chemistry degree courses.
Definition This allows greater flexibility in subject choice and accommodates students
studying on different degree paths e.g. B.Sc. Chemistry or MChem. Modules
Transcript – this is your record of
also break a subject into discrete, easily assimilated elements. They have the
achievement at university. Normally it
advantage of spreading assessment over the academic year, but they can also
will consist of details of each module or
course you have taken, and an indica- tempt you to avoid certain difficult subjects or to feel that you can forget about
tion of the grade or mark achieved. In a topic once the module is finished.
the UK your transcript will also show
your final (honours) classification, that
is: first class, upper second class (2.1), Key Point You should select your modules with care, mindful
lower second class (2.2), third class or of potential degree options and how your transcript and CV will
unclassified (note: some UK universi- appear to a prospective employer. If you feel you need advice,
ties do not differentiate second class
consult your personal tutor or study advisor.
degrees).
As you move between levels of the university system, you will be expected to
Aiming high – your goal should be have passed a certain number of modules, as detailed in the progression criteria.
to perform at your highest possible These may be expressed using a credit point system. Students are normally
level and not simply to fulfil the mini- allowed two attempts at each module exam and the resits often take place at the
mum criteria for progression. This will end of the summer vacation. If a student does not pass at the second attempt,
lay sound foundations for your later they may be asked to ‘carry’ the subject in a subsequent year, and in severe
studies. Remember too that a future cases of multiple failure, they may be asked to re-take the whole year or even
employer might ask to see your aca- leave the course. Consequently, it is worth finding out about these aspects of
demic transcript, which will detail all your degree. They are usually published in course handbooks.
your module grades including any fails/ You are unlikely to have reached this stage in your education without being
resits, and will not just state your final exposed to the examination process. You may not enjoy being assessed, but
degree classification. you probably want to do well in your course. It is therefore important to under-
stand why and how you are being tested. Identifying and improving the skills
required for exam success will allow you to perform to the best of your ability.
Seeking advice on module options –
most universities provide students Assessed coursework
with access to a staff member who acts There is a component of assessed coursework in many modules. This often
as an impartial advisor on curriculum tests specific skills, and may require you to demonstrate thinking at deeper
matters. Careers service staff may also levels (Table 62.1). The common types of coursework assessment are covered
offer valuable guidance. Make sure you at various points in this book:
take advantage of this by setting up and
attending meeting with relevant per- ●● practical exercises (Chapters 1–40);
sonnel. Senior students can also be a ●● essays (Chapters 68 and 69);
useful source of advice but always com-
pare their opinions with those of staff. ●● numerical problems (Chapter 51);
●● data analysis (Chapters 51–54);
●● poster and spoken presentations (Chapters 66 and 67);
Avoiding plagiarism – this is a key
●● literature surveys and reviews (Chapter 71);
issue for assessed coursework – see
p. 386 for a definition and Chapter 4 ●● project work (Chapters 7 and 69);
for appropriate methods of referring
●● problem-based learning (Box 64.1).
to the ideas and results of others using
citation. At the start of each year or module, read the course handbook or module
guide carefully to find out when any assessed work needs to be submitted. Note

Assignments and exams
Box 64.1 Problem-based learning (PBL)
In this teaching method, you are likely to be presented 4. Having carried out the necessary research, you should
with a ‘real world’ problem or issue, often working within review the information that has been obtained. As
a team. As you tackle the problem, you are expected to a result, new issues may need to be explored and,
gain factual knowledge, develop skills and exercise crit- where appropriate, allocated to group members.
ical thinking (Chapter 42). Because there is a direct and
5. You will be asked to produce an outcome, such as a
relevant context for your work, and because you have to
report, diagnosis, seminar presentation or poster. An
employ active learning techniques, the knowledge and
outline structure will be required, and for groups, fur-
skills you gain are likely to be more readily assimilated
ther allocation of tasks will be needed to accomplish
and remembered. This approach also more closely mim-
this outcome.
ics workplace practices. PBL usually proceeds as follows:
If asked to carry out PBL as part of your course, it is impor-
1. You are presented with a problem (e.g. a case study,
tant to get off to a good start. At first, the problem may
a hypothetical patient, a topical issue).
seem unfamiliar. However, once you become involved in
2. You consider what issues and topics you need to the work, you will quickly gain confidence. If working as
research, by discussion with others if necessary. You part of a group, make sure that your group meets as early
may need to identify where relevant resources can be as possible, that you attend all sessions and that you do
found (Chapters 41– 44). the necessary background reading. When working in a
team, a degree of self-awareness is necessary regarding
3. You then need to rank the issues and topics in impor-
your ‘natural’ role in group situations (Table 59.1). Vari-
tance, allocating tasks to group members, if appro-
ous methods are used for grading PBL, including written,
priate. A structured approach is an important aspect
oral and poster presentations, and this may involve self-
of PBL.
assessment and/or peer marking.
Summative and formative aspects

relevant dates in your diary, and use this information to plan your work. Take
of assessment – many assessments
special note if deadlines for different modules clash, or if they coincide with
have a formative component, providing
social or sporting commitments.
feedback that is aimed at helping you to
improve, as well as a summative com-
ponent, with a mark that contributes to
Key Point If, for some valid reason (e.g. illness), you will be
your final grade.
late with an assessment, speak to your tutors as soon as pos-
sible. They may be able to take extenuating circumstances into
account by not applying a marking penalty. They will let you
Definitions know what paperwork you may be required to submit to sup-
port your claim.
Terms with specialised meaning in uni-
versity exam contexts:
Diet – a period during which a number Summative exams – general points
of exams take place, according to a pub-
lished timetable. Summative exams (p. 555) normally involve you answering questions without
Invigilator – a person whose role is to being able to consult other students or your notes. Invigilators are present to
ensume an exam is conducted accord- ensure appropriate conduct, but departmental representatives may be present
ing to the regulations. for some of the exam. Their role is to sort out any subject-related problems,
Paper – usually, a set of questions for a so if you think something is wrong, ask at the earliest opportunity. It is not
single subject. unknown for parts of questions to be omitted in error, or for double meanings
Question – an element of an exam to arise, for example.
paper, often couched in terms of an
Planning
instruction rather than a true question,
and often with the marks available When preparing for an exam, make a checklist of the items you will need (see
stated at the end. p. 567). On the day of the exam, give yourself sufficient time to arrive at the
Rubric – the set of instructions at the correct room, without the risk of being late. Double-check the times and places
start of an exam paper. of your exams, both well before the exam, and also on arrival. If you arrive at

the exam venue early, you can always rectify a mistake if you find you have
gone to the wrong place.
Tackling the paper

Begin by reading the instructions at the top of the exam paper carefully, so
that you do not make any errors based on lack of understanding of the exam
structure. Make sure that you confirm:
●● how many questions are set;
●● how many must be answered;
●● whether the paper is divided into sections;
●● whether any parts are compulsory;
●● what each question/section is worth, as a proportion of the total mark;
●● whether different questions should be answered in different books.
Do not be tempted to spend too long on any one question or section: the return
in terms of marks will not justify the loss of time from other questions (see
Fig. 64.1). Take the first few minutes to read the paper and plan your strategy,
before you begin writing. Do not be put off by those who begin immediately; it
is almost certain they are producing unplanned work of a poor standard.
Underline the key phrases in the instructions, to reinforce their message.
Next, read through the set of questions. If there is a choice, decide on those
efficient use
questions to be answered and decide on the order in which you will tackle
of time them. Quickly prepare a timetable which takes into account the amount of time
use of time not required to complete each section and which reflects the allocation of marks –
cost-effective
there is little point in spending one-quarter of the exam period on a question
Cumulative mark
worth only 5% of the total marks. Use the exam paper to mark the sequence in
which the questions will be answered and write the finishing times alongside;
refer to this timetable during the exam to keep yourself on course.
Reviewing your answers

At the end of the exam, you should allow some time to check through your
script. Make sure your name and/or ID number is on each exam book as
Time required and on all other sheets of paper, including graph paper, even if securely
attached to your script, as it is in your interest to ensure that your work does
Fig. 64.1 Potential exam marks as a func- not go astray.
tion of time. The marks awarded in a single
answer will follow the law of diminishing
returns – it will be far more difficult to Key Point Never leave any exam early. Most exams assess
achieve the final 25% of the available marks work carried out over several months within a time period of
than the initial 25%. Do not spend too long 2–3 h and there is always something constructive you can do
on any one question. with the remaining time to improve your script.
Special considerations for different types of exam questions

Using the question paper – unless this Essay questions
is specifically forbidden, you should
write on the question paper to plan
Essay questions let examiners test the depth of your comprehension and under-
your strategy, keep to time and organ-
standing as well as your recall of facts. Essay questions give you plenty of
ise answers.
scope to show what you know. They suit those with a good grasp of principles
but who perhaps have less ability to recall individual details.

Before you tackle a particular question, you must be sure of what is

Checking your exam answers – look
required in your answer. Ask yourself ‘What is the examiner looking for in
for:
this particular question?’ and then set about providing a relevant answer.
●● errors of fact;
●● missing information;
Consider each word in the question and highlight, underline or circle the key
●● grammatical and spelling errors;
words. Make sure you know the meaning of the terms given in Table 69.1 so
●● errors of scale and units; that you can provide the appropriate information, where necessary. Spend
●● errors in calculations. some time developing a structure for your writing (see Chapter 68). Refer
back to the question frequently as you write, to confirm that you are keeping
to the subject matter. Box 64.2 gives advice on writing essays under exam
conditions.
Adopting different tactics according
It is usually a good idea to begin with the question that you are most
to the exam – you should adjust your
confident about. This will reassure you before tackling more difficult parts
exam strategy (and revision methods)
of the paper. If you run out of time, write in note form. Examiners are usu-
to allow for the differences in question
ally understanding, as long as the main components of the question have been
types used in each exam paper.
addressed and the intended structure of the answer is clear. Common reasons
for poor exam answers in essay-style questions are listed in Box 64.3. Try to
avoid making these mistakes.
Penalties for guessing – if there is a
penalty for incorrect answers in a multi-
ple choice test, the best strategy is not to Multiple-choice and short-answer questions
answer questions when you know your Multiple-choice questions (MCQs) and short-answer questions (SAQs) are gen-
answer is a complete guess. Depending erally used to test the breadth and detail of your knowledge. The various styles
on the penalty, it may be beneficial to that can be encompassed within the SAQ format allow for more demanding
guess if you can narrow the choice down questions than MCQs, which may emphasise memory work and specific factual
to two options (but beware false or irrel- information.
evant alternatives). However, if there are A good approach for MCQ papers is as follows:
no such penalties, then you should pro-
vide an answer to all questions.
1. First trawl: read through all of the questions fairly rapidly, noting the
‘correct’ answer for those you can attempt immediately.
Box 64.2 Writing under exam conditions
Make sure you go into an exam with a strategy for man- ●● Use standard abbreviations to save time repeating
aging the available time. text but always explain them at the first point of use
(e.g. HPLC, high performance liquid chromatography).
●● Allocate some time (say 5% of the total) to consider
which questions to answer and in which order. ●● Consider speed of writing and neatness – especially
when selecting the type of pen to use – ballpoint pens
●● Share the rest of the time among the questions,
are fastest, but they can tend to smudge. You can only
according to the marks available. Aim to optimise the
gain marks if the examiner can read your script.
marks obtained. A potentially good answer should be
allocated slightly more time than one you do not feel ●● Keep your answer simple and to the point, with clear
so happy about. However, do not concentrate on any explanations of your reasoning.
one answer (see Fig. 64.1).
Make sure your answer is relevant.
●● For each question divide the time into planning, writ-
●● Do not include irrelevant facts just because you mem-
ing and revision phases (see p. 598).
orised them during revision, as this may do you more
Employ time-saving techniques as much as possible: harm than good. You must answer the specific ques-
tion that has been set.
●● Use spider diagrams (Fig. 68.2) or mind maps
(Fig. 61.2) to organise and plan your answer. ●● Remember that time taken to write irrelevant material
is time lost from another question.
●● Use diagrams and tables to save time in making diffi-
cult and lengthy explanations, but make sure you refer
to each one in the text.

Box 64.3 Reasons for poor exam answers to essay-style questions
The following are reasons that lecturers often cite when ●● Illegible handwriting.
they give low marks for essay answers:
●● Poor English, such that facts and ideas are not
●● Not answering the exact question set. Either failing expressed clearly.
to recognise the specialist terms used in the question, ●● Lack of logic or structure to the answer.
failing to demonstrate an understanding of the terms
by not providing definitions, failing to carry out the ●● Factual errors, indicating poor note-taking, poor revi-
precise instruction in a question, or failing to address sion or poor recall.
all aspects of the question. ●● Failing to correct obvious mistakes by re-reading an
●● Running out of time. Not matching the time allocated answer before submitting the script.
to the extent of the answer. Frequently, this results in At higher levels, the following aspects are especially
spending too long on one question and not enough important:
on the others, or even failing to complete the paper.
●● Not providing enough in-depth information.
●● Not answering all parts of a multiple-part question, or
not recognising that one part (perhaps involving more ●● Providing a descriptive rather than an analytical
complex ideas) may carry more marks than another. answer – focusing on facts, rather than deeper aspects
of a topic (see Table 62.1).
●● Failing to provide evidence to support an answer. For-
getting to state the ‘obvious’ – either basic facts or ●● Not setting a problem in context, or not demonstrat-
definitions. ing a wider understanding of the topic. However,
make sure you do not overdo this, or you may risk not
●● Failing to illustrate an answer appropriately , either
answering the question set.
by not including a relevant diagram, or by providing a
diagram that does not aid communication, or by not ●● Not giving enough evidence of reading around the
including relevant examples. subject. Wider reading can be demonstrated by quot-
ing relevant papers and reviews and by giving author
●● Incomplete answer(s). Failing to answer appropriately
names and dates of publication.
owing to lack of knowledge.
●● Not considering both sides of a topic/debate, or not
●● Providing irrelevant evidence to support an answer.
arriving at a conclusion.
‘Waffling’ to fill space.
2. Second trawl: go through the questions again, checking your original

answers and answering any other straight forward questions.
3. Third trawl: now tackle the difficult questions and those that require
longer to answer (e.g. those based on numerical problems).
4. Final review: look again at all your answers, check for obvious mistakes
When unsure of an answer, the first stage is to rule out options that are
Answer the question as requested – clearly absurd or have obviously been placed there to distract you. Next, look-
this is true for all questions, but ing at the remaining options, can you judge between contrasting pairs with
especially important for SAQs. If the alternative answers? Logically, both cannot be correct, so you should see if you
question asks for a diagram, make sure can rule one of the pair out. Watch out, however, in case both may be irrelevant
you provide one; if it asks for a speci- to the answer. If the question involves a calculation, try to work this out inde-
fied number of aspects of a topic, try to pendently from the answers, so you are not influenced by them.
list this number of points; if there are In SAQ papers, there may be a choice of questions. Choose your options
two or more parts, provide appropriate carefully – it may be better to gain half marks for a correct answer to half a
answers to all aspects. This may seem question, than to provide a largely irrelevant answer that apparently covers the
obvious, but many marks are lost for whole question but lacks the necessary detail. For the SAQ form of question,
not following instructions. few if any marks are given for writing style. Think in ‘bullet points’ and list the
crucial points only. The time for answering SAQ questions may be tight, so get

down to work fast, starting with answers that demand remembered facts. Stick
Typical layout for an extended match-
to your timetable by moving on to the next question as soon as possible. Stra-
ing items (EMI) question
tegically, it is probably better to get part-marks for the full number of questions
1. Description of topic or theme – than good marks for only a few.
use this heading to orientate your
thoughts when answering. Extended matching items (EMI)
2. Instructions – this will explain how EMIs, sometimes called extended matching sets, or extended matching ques-
you should answer and the weight- tions, are a more complex and challenging version of the multiple-choice for-
ing of marks among the questions. mat, where a pool of answers is offered that may be correct in several cases
If all the questions are similar, this or none at all. Formats differ, but the questions often relate to a complex sce-
may appear at the head of the exam nario outlined as a ’vignette’. EMIs are designed to test not only your detailed
paper, if not, read each instruction knowledge but also your deeper understanding and ability to apply this, and
carefully, as the rules and maiks are commonly used in disciplines where professional judgements on complex
given may change. issues must be made.
3. A series of options – you will have Exams with an EMI component require you to read carefully, quickly and
to select among these to answer with understanding, distinguish between relevant and irrelevant information
the questions which follow. Some and make accurate connections between statements. As with MCQs, you will
options may be correct more than require a broad knowledge across your subject, but also a deeper and sometimes
once, some not at all, and you may quite detailed understanding. An idea of the sorts of topics that might be cov-
or may not be told how many are ered should be apparent from the learning outcomes and case studies covered in
correct in each case. Each time you the course. Look carefully at any example EMIs or past papers that are offered,
answer a question you will have to as this will help you to develop a strategy for answering.
go through the list carefully and When answering an EMI paper, first look quickly through the topics and
see which apply – slow readers select those that most suit your knowledge and revision. Plan to answer those
beware! you are most confident about first. Then, for each individual EMI:
4. A scenario (or ‘vignette’) – this is 1. Scan-read the whole question quickly to get a feel for its scope.
a short paragraph that describes 2. Read both the scenario and the questions more carefully, and highlight
the background to the questions. key terms. Some people find it helpful next to try to answer the questions
Details here will have a big effect without reference to the options as they will not be put off by distracters.
on the answers you give, so you
will need to consider the wording 3. Work through the list of options ruling them ‘in’ or ‘out’ for each spe-
very carefully before you answer It cific question – you should be left with a shortlist of possible answers.
might be a good idea to highlight 4. Select the final answers you wish to give, with reference to the instruc-
key words to keep them in your tions or rules provided.
mind.
5. Make sure you are keeping up with the necessary pace by setting a
5. The questions – these may be target time for completion of each topic. Try to leave some time in your
grouped, and may develop in com- planning to allow you to review and check your answers (p. 562).
plexity from those demanding a
simple factual answer to those
You will need to be particularly well focused and work quickly and decisively –
requiring a fair amount of reflection
time will often be a constraint in EMI exams. If there are no penalty marks,
and judgement.
ensure you give a response to each question.
Practical and information-processing exams

The prospect of a practical or information-processing exam may cause you
more concern than a theory exam. This may be due to a limited experience of
practical examinations, or to the fact that practical and observational skills are
tested, as well as recall, description and analysis of factual information. Your
first thoughts may be that it is not possible to prepare for such exams but, in
fact, you can improve your performance by mastering the various practical
techniques described in this book.

You may be allowed to take your laboratory reports and other texts into
Examples These are principal types
the practical exam. Do not assume that this is a soft option, or that revision is
of questions you are likely to encounter
unnecessary: you will not have time to read large sections of your reports or
in a practical or information-processing
to familiarise yourself with basic principles, etc. The main advantage of ‘open
exam:
book’ exams is that you can check specific details of methodology, reducing
Manipulative exercises Often based your reliance on memory, provided you know your way around your practical
on work carried out during your prac- manual and notes. In all other respects, your revision and preparation for such
tical course. Tests dexterity, specific
exams should be similar to theory exams. Make sure you are familiar with all of
techniques (e.g. sample injection in gas
chromatography).
the practical exercises, including any work carried out in class by your partner
(since exams are assessed on individual performance). If necessary, check with
‘Spot’ tests Short questions requiring
the teaching staff to see whether you can be given access to the laboratory to
an identification, or brief descriptive
notes on a specific item (e.g. define the
complete any exercises that you have missed.
operation of a hollow cathode lamp). At the outset of the practical exam, determine or decide on the order in
Tests knowledge of seen material or which you will tackle the questions. A question in the latter half of the paper
ability to transfer this to a new example. may need to be started early on in the exam period (e.g. a chemical reflux
Calculations May include the prepa- (Chapter 17) requiring 2.h incubation in a 3.h exam). Such questions are
ration of aqueous solutions at particu- included to test your forward planning and time management skills. You may
lar concentrations (Chapter 11) and need to make additional decisions on the allocation of material; e.g. if you need
statistical exercises (Chapter 54). Tests to run a melting point, IR spectrum and a UV/visible spectrum, determine how
numeracy. much compound may be required for each before starting, particularly if some
Data analyses May include the prepa- of the compound is required to answer another question.
ration and interpretation of graphs Make sure you explain your choice of apparatus and experimental design.
(Chapter 49) and numerical informa- Calculations should be set out in a stepwise manner, so that credit can be given,
tion, from data either obtained during even if the final answer is incorrect (see p. 468). If there are any questions that
the exam or provided by the examiner. rely on recall of factual information and you are unable to remember specific
Tests problem-solving skills. details, make sure that you describe the item fully, so that you gain credit for
Chemical synthesis May include observational skills. Alternatively, leave a gap and return to the question at a
details of starting reagents and later stage.
require possible likely products to be
suggested.
Chemical structure elucidation A Oral exams and interviews
series of spectra are provided (IR, An oral interview is sometimes a part of final degree exams, representing a
nmr) which require interpretation and chance for the external examiner(s) to meet individual students and to test
identification of the probable chemical their abilities directly and interactively. In some departments, orals are used to
compound. validate the exam standard or to test students on the borderline between exam
grades. Sometimes an interview may form part of an assessment, as with pro-
ject work or posters. This type of exam is often intimidating – many students
Terminology – the oral exams are say they do not know how to revise for an oral – and many candidates worry
sometimes known simply as ‘orals’ or, that they will be so nervous they will not be able to do themselves justice.
borrowing Latin, as ‘viva voce’ (by the Preparation is just as important for oral exams as it is for written exams:
living voice) exams or ‘vivas’. ●● Think about your earlier performances – if the oral follows written
papers, it may be that you will be asked about questions you did not do so
well on. These topics should be revised thoroughly. Be prepared to say how
you would approach the questions if given a second chance.
●● Read up a little about the examiner – he or she may focus their questions
in their area of expertise.
●● Get used to giving spoken answers – it is often difficult to transfer between
written and spoken modes. Write down a few questions and get a friend to
ask you them, possibly with unscripted follow-up queries.
●● Research and think about topical issues in your subject area – some
examiners will feel this reflects how interested you are in your subject.

Your conduct during the oral exam is important, too:

Allow yourself to relax in oral
exams – external examiners are expe-
●● Arrive promptly and wear reasonably smart clothing. Not to do either
rienced at putting students at ease. They
might be considered disrespectful by the examiner.
will start by asking ‘simple-to-answer’
questions, such as what modules you ●● Take your time before answering questions. Even if you think you know
did, how your project research went, the answer immediately, take a little while to check mentally whether you
and what your career aspirations are. have considered all angles. A considered, logical approach will be more
Look on the external as a friend rather impressive than a quick but ill-considered response.
than a foe.
●● Start answers with the basics, then develop into deeper aspects. There
may be both surface and deeper aspects to a topic and more credit will be
given to a student who puts the latter in context.
●● When your answer is finished, stop speaking. A short, crisp answer is
better than a rambling one.
Creating an exam action list – knowing ●● If you do not know the answer, say so. To waffle and talk about irrelevant
that you have prepared well, checked material is more damaging than admitting that you do not know.
everything on your list of actions and ●● Make sure your answer is balanced. Talk about the evidence and opinions
gathered together all you need for an on both sides of a contentious issue.
exam will improve your confidence and
reduce anxiety. Your list might include: ●● Do not disagree violently with the examiner. Politely put your point
●● Time, date and place of the exam. of view, detailing the evidence behind it. Examiners will be impressed
●● Travel arrangements to exam hall. by students who know their own mind and subject area. However, they
●● Double-check module handbooks will expect you to support a position at odds with the conventional
and past papers for exam structure. viewpoint.
●● Think through use of time and exam
strategy. ●● Finally, be positive and enthusiastic about your topic.
●● Identify a quiet place near the exam
hall to carry out a last-minute check
on key knowledge (e.g. formulae, Counteracting anxiety before and during exams
definitions, diagram labels).
●● Ensure you have all the items you Adverse effects of anxiety need to be overcome by anticipation and preparation
wish to take to the exam, e.g. well in advance (Box 64.4). Exams, with their tight time limits, are especially
– pens, pencils (with sharpener and stressful for perfectionists. To counteract this tendency, focus on the following
eraser); points during the exam:
– ruler;
– correction fluid; ●● Do not expect to produce a perfect essay – this will not be possible in the
– c alculator (allowable type), if time available.
required;
●● Do not spend too long planning your answer – once you have an outline
– sweets and drink, if allowed;
plan, get started.
– tissues
– watch or clock; ●● Do not spend too much time on the initial parts of an answer, at the
– ID card; expense of the main message.
– texts and/or notes, if an open book ●● Concentrate on getting all of the basic points across – examiners often
exam;
allocating marks for the main points first, before allocating extra marks for
– lucky charm/mascot. the detail and depth.
●● Lay out clothes (if exam is early in
the morning). ●● Do not be obsessed with neatness, either in handwriting, or in the diagrams
●● Set alarm(s) and/or ask a friend or you draw, but make sure your answers are legible.
family member to check you are
awake on time. ●● Do not worry if you forget something. You can not be expected to know
everything. Most marking schemes give a first class grade to work that
misses out on up to 30% of the marks available.

Box 64.4 Strategies for combating the symptoms of exam anxiety
●● Sleeplessness – this is common and does little harm it may be a good idea to get up early for a few days
in the short term. Get up, have a snack, do some light beforehand.
reading or other activity, then return to bed. Avoid caf-
●● Blind panic during an exam – explain how you feel to
feine (e.g. tea, coffee and cola) for several hours before
an invigilator. Ask to go for a supervised walk outside.
going to bed.
Do some relaxation exercises (see below), then return
●● Lack of appetite – again commonplace. Eat what you to your work. If you are having problems with a spe-
can, and take sugary sweets into the exam to keep cific question, it may be appropriate to speak to the
energy levels up in case you become tired. departmental representative at the exam, to check that
you are not misinterpreting the question.
●● Fear of the unknown – it can be a good idea to visit the
exam room beforehand, so you can become familiar ●● Feeling tense – shut your eyes, take several slow,
with the location. By working through the points given deep breaths, do some stretching and relaxing muscle
in the exam action list on p. 567, you will be confident movements. During exams, it can be a good idea to do
that nothing has been left out. this between questions, and possibly to have a com-
plete rest for a minute or so. Before exams, try some
●● Lack of preparation – the remedy, of course, is to adopt
exercise activity or escape temporarily from your wor-
an effective approach to your revision (Chapter 63).
ries by watching TV or a movie.
●● Worries about timekeeping – get a reliable alarm clock
●● Running out of time – do not panic when the invigilator
or a new battery for an old one. Arrange for an alarm
says ‘five minutes left’. It is surprising how much you can
phone call; ask a friend or relative to make sure you
write in this time. Write note-style answers or state the
are awake on time. Make reliable travel arrangements,
areas you would have covered: you may get some credit.
to arrive on time. If your exam is early in the morning,
Key Point Everyone worries about exams. Anxiety is a per-

fectly natural feeling. It works to your advantage, as it helps pro-
After the exam – try to avoid becom-
vide motivation and the adrenaline that can help you ‘raise your
ing involved in prolonged analyses with
game’ on the day.
other students over the ‘ideal’ answers
to the questions; after all, it is too late
to change anything at this stage. Go
for a walk, watch TV for a while, or do
There is a lot to be said for treating exams as a game. After all, they are artificial
something else that helps you relax, so
situations contrived to ensure that large numbers of candidates can be assessed
that you are ready to face the next exam
together, with little risk of cheating. They have conventions and rules, just like
with confidence.
games. If you understand the rationale behind them and follow the rules, this
will aid your performance.

Acres, D. (1998) Passing Exams Without Anxiety: How to O’Brien, D. (2007) How to Pass Exams: Accelerate Your
Get Organised, be Prepared and Feel Confident of Suc- Learning - Memorise Key Facts - Revise Effectively, 2nd
cess. How to Books, London. edn. Baird, Winchester.
McMillan, K.M and Weyers, J.D.B. (2011) How to Suc- [Many universities host study skills websites that cover
ceed in Exams and Assessments, 2nd edn. Prentice Hall, exam technique; these can be found using ‘study skills’,
London. ‘revision’ or ‘exams’ as key words in a search engine.]

Study exercises
64.1 Analyse your past performances. Think back some time to discussing your revision notes with
to past exams and any feedback you received a colleague. Try to learn from his or her approach.
from them. How might you improve your per- Discuss any issues you do not agree upon.
formance? Consider ways in which you might
64.3 Plan your exam tactics. Find out from your module
approach the forthcoming exam differently. If
handbook or past papers what the format of each
you have kept past papers and answers to con-
paper will be. Confirm this if necessary with staff.
tinuous assessment exercises, look at any spe-
Decide how you will tackle each paper, allocating
cific comments your lecturers may have made.
time to each section and to each question within
64.2 Share revision notes with other students. Make a the sections (see p. 598). Write a personal checklist
revision plan (see pp. 535, 557) and then allocate of requirements for the exam (see p. 567).

65 Preparing your curriculum vitae
Many students only think about their curriculum vitae immediately before
Definition applying for a job. However this should not be the case. Thinking and drafting
your CV at an early stage of your degree course is important. You can always
Curriculum vitae (or CV for short) – a
add more (and revise it) as your degree progresses. There are four main reasons
Latin phrase that means ‘the course
why this can be valuable:
your life has taken’.
1. Considering your CV and how it will look to a future employer will help
you think more deeply about the direction and value of your academic
studies.
Personal development planning (PDP)
and your CV – many PDP schemes 2. Creating a draft CV will prompt you to assess your skills and personal
(p. 529) also include an element of qualities and how these fit into your career aspirations.
career planning that may involve creat- 3. Your CV can be used as a record of all the relevant things you have done
ing a draft or generic CV. The PDP pro- at university and then, later, will help you communicate these to a potential
cess can help you improve the structure employer.
and content of your CV, and the lan-
guage you use within it.
4. Your developing CV can be used when you apply for vacation or part-time
employment.
Key Point Developing your skills and qualities needs to be

treated as a long-term project. It makes sense to think early
about your career aspirations so that you can make the most of
opportunities to build up relevant experience. A good focus for
such thoughts is your developing curriculum vitae, so it is useful
to work on this from a very early stage.
Skills and personal qualities

Understanding skills and qualities – it
may be helpful to think about how the Skills (sometimes called competences) are generally what you have learned
skills and qualities in Tables 58.1 and to do and have improved with practice. Table 1.1 summarises some important
65.1 apply to particular activities during skills for bioscientists. This list might seem quite daunting, but your tutors
your studies, since this will give them a will have designed your courses to give you plenty of opportunities to develop
greater relevance. your expertise. Personal qualities, on the other hand, are predominantly innate.
Examples include honesty, determination and thoroughness (Table 65.1). These
qualities need not remain static, however, and can be developed or changed
according to your experiences. By consciously deciding to take on new chal-
Focusing on evidence – it is important lenges and responsibilities, not only can you develop your personal qualities,
to be able to provide specific informa- but you can also provide supporting evidence for your CV.
tion that will back up the claims you Personal qualities and skills are interrelated because your personal qual-
make under the ‘skills and personal ities can influence the skills you gain. For example, you may become highly
qualities’ and other sections of your CV. proficient at a skill requiring manual dexterity if you are particularly adept with
A potential employer will be interested your hands. Being able to transfer your skills is highly important (Chapter 58) –
in your level of competence (what you many employers take a long-term view and look for evidence of the adaptability
can actually do) and in situations where that will allow you to be a flexible employee and one who will continue to
you have used a skill or demonstrated develop skills.
a particular quality. These aspects can
also be mentioned in your covering let- Developing your curriculum vitae
ter or at interview.
The initial stage involves making an audit of the skills and qualities you already
have, and thinking about those you might need to develop. Tables 58.1 and 65.1
could form a basis of this self-appraisal. Assessing your skills may be easier

Preparing your curriculum vitae
Table 65.1 Some positive personal qualities. than critically analysing your personal characteristics. In judging your qualities,
try to take a positive view and avoid being overly modest. It is important to
Adaptability consider personal qualities in a specific context, e.g. ‘I have shown that I am
Conscientiousness
Curiosity trustworthy, acting as treasurer for the Chemical Society’, as this evidence will
Determination form a vital part of your CV and job applications.
Drive If you can identify gaps in your skills, or qualities that you would like to
Energy
Enthusiasm develop, especially in relation to the needs of your intended career, the next
Fitness and health step is to think about ways of improving them. This will be reasonably easy
Flexible approach
Honesty
in some cases, but may require some creative thinking in others. A relatively
Innovation simple example would be if you decided to learn a new language or to main-
Integrity tain one you learned at school. There are likely to be many local college and
Leadership
Logical approach university courses dealing with foreign languages at many different levels, so it
Motivation would be a straightforward matter to join one of these. A rather more difficult
Patience case might be if you wished to demonstrate ‘responsibility’, because there are
Performance under stress
Perseverance no courses available on this. One route to demonstrate this quality might be to
Prudence put yourself up for election as an officer in a student society or club; another
Quickness of thought could be to take a leading role in a relevant activity within your community
Seeing others’ viewpoints
Self-confidence (e.g. voluntary work). If you already take part in activities like these, your CV
Self-discipline should relate them to this context.
Sense of purpose
Shrewd judgement
Social skills (sociability)
Taking initiative Basic CV structures and their presentation
Tenacity
Tidiness Box 65.1 illustrates the typical parts of a CV and explains the purpose of each
Thoroughness part. Employers are more likely to take notice of a well-organised and well
Tolerance presented CV, in contrast to one that is difficult to read and assimilate. They
Unemotional approach
Willingness to take on challenges will expect it to be concise, complete and accurate. There are many ways
of presenting information in a CV, and you will be assessed partly on your
choices.
●● Order. There is some flexibility as to the order in which you can present
Seeing yourself as others see you – you
the different parts (see Box 65.1). A chronological approach within sections
may not recognise all of your personal
helps employers gain a picture of your experience.
qualities and you may need someone
else to give you an honest appraisal. ●● Personality and ‘colour’. Make your CV different by avoiding standard or
This could be anyone whose opinion dull phrasing. Try not to focus solely on academic aspects: you will proba-
you value: a friend, a member of your bly work in a team and the social aspects of teamwork will be enhanced by
family, a tutor or a careers adviser. your outside interests. However, make sure that the reader does not get the
impression that these interests dominate your life.
●● Style. Your CV should reflect your personality, but not in such a way that
Setting your own agenda – you have it indicates too idiosyncratic an approach. It is probably better to be formal
the capability to widen your experience in both language and presentation, as flippant or chatty expressions will not
and to demonstrate relevant personal be well received.
qualities through both curricular and
extracurricular activities. ●● Neatness. Producing a well-presented, word processed CV is very impor-
tant. Use a laser quality printer and good quality paper; avoid poor quality
photocopying at all costs.
Paying attention to the quality of ●● Layout. Use headings for different aspects, such as personal details, educa-
your CV – your potential employer tion, etc. Emphasise words (e.g. with capitals, bold, italics or underlining)
will regard your CV as an example of sparingly and with the primary aim of making the structure clearer. Remem-
your very best work and will not be ber that careful use of white space is important in design.
impressed if it is full of mistakes or
badly presented, especially if you claim
●● Grammar and proofreading. Look at your CV carefully before you submit
‘good written communication’ as a skill!
it, as sloppy errors give a very poor impression. Even if you use a spell-
checker, some errors may creep in. Ask someone whom you regard as a

Box 65.1 The structure and components of a typical CV and covering letter
There is no right or wrong way to write a CV, and no sin- employers may ask for three). It is usual to include
gle format applies. It is probably best to avoid software your personal tutor or course leader at university
templates and CV ‘wizards’ as they can create a bland, (who among other things, will verify your marks), plus
standardised result, rather than something that demon- another person – perhaps a current or former employer,
strates your individuality. or someone who runs a club or society and who knows
It is important to focus on the relevant issues, keeping your personal interests and activities. Unless you have
your writing concise, well-structured and well-presented. kept in touch with a particular teacher since starting uni-
You should include the following, with appropriate sub- versity, it is probably best to choose current contacts,
headings, generally in the order given below: rather than those from your previous education.
1. Personal details. This section must include your full Some other points to consider:
name and date of birth, your address (both home and ●● Try to avoid jargon and over-complicated phrases in
term-time, with dates, if appropriate) and a contact your CV: aim for direct, active words and phrases (see
telephone number at each address. If you have an Box 68.1).
email account, you might also include this. You need
only mention sex if your name could be either male ●● Most employers will expect your CV to be word-
or female. processed (and spell-checked). Errors in style, gram-
mar and presentation will count against you, so be
2. Education. Choose either chronological order, or
sure to check through your final version (and ask a
reverse chronological order and make sure you take
reliable person to second-check it for you).
the same approach in all other sections. Give edu-
cational institutions and dates (month, year) and ●● Aim for a maximum length of two pages, printed
provide more detail for your degree course than for single-sided on A4 paper, using a ‘formal’ font (e.g.
your previous education. Remember to mention any Times Roman or Arial) of no less than 12 point for the
prizes, scholarships or other academic achievements. main text. Always print onto good’ quality white paper.
Include your overall mark for the most recent year of Avoid fussy use of colour, borders or fonts.
your course, if it seems appropriate. Make sure you ●● Do not try to cram in too much detail. Use a clear and suc-
explain any gap years. cinct approach with short sentences and lists to improve
3. Work experience. Include all temporary, part-time, ‘readability’ and create structure. Remember that your
full-time or voluntary jobs. Details include dates, aim is to catch the eye of your potential employer, who
employer, job title and major duties involved. may have many applications to work through.
4. Skills and personal qualities. Tables 58.1 and 65.1 give ●● It is polite to check that people are willing to act as a
examples of the aspects you might include. Empha- referee for you and to provide them with an up-to-date
sise your strengths, and tailor this section to the spe- copy of your CV.
cific requirements of the post (the ‘job description’):
for example, the practical skills you have gained dur- Your single-page covering letter should have four major
ing your degree studies if the post is a chemical one, components:
but concentrate on generic transferable skills and 1. Letterhead. Include your contact details, the recipi-
personal qualities for other jobs. Provide supporting ent’s name and title (if known) and address, plus any
evidence for your statements in all cases. job reference number.
5. Interests and activities. This is an opportunity to bring 2. Introductory paragraph. Explain who you are and
out the positive aspects of your personality, and explain state the post for which you are applying.
their relevance to the post you are applying for. Aim to
3. Main message. This is your opportunity to sell yourself
keep this section short, or it may seem that your social
to a potential employer, highlighting particular attrib-
life is more important than your education and work
utes and experience. Keep it to three or four sentences
experience. Include up to four separate items, and pro-
at the most and relate it to the particular skills and
vide sufficient detail to highlight the positive aspects of
qualities demanded in the job or person specification.
your interests (e.g. positions of responsibility, working
with others, communication, etc.). Use sections 4 and 5 4. Concluding paragraph. A brief statement that you
to demonstrate that you have the necessary attributes look forward to hearing the outcome of your applica-
to fulfil the major requirements of the post. tion is sufficient.
6. Referees. Include the names (and titles), job descrip- Finally, add either ‘Yours sincerely’ (where the recipient’s
tions, full postal addresses, contact telephone num- name is known) or ‘Yours faithfully’ (in a letter beginning
bers and email addresses of two referees (rarely, some ‘Dear Sir or Madam’) and then end with your signature.

reliable proofreader to comment on it (many tutors will do this, if asked in

Creating a generic CV – as you may
advance).
apply for several jobs, it is useful to
construct a CV in electronic format (e.g. ●● Relevance. If you can, slant your CV towards the job description and the
as a Word file) which includes all infor- qualifications required (see below). Make sure you provide evidence to back
mation of potential relevance. This can up your assertions about skills, qualities and experience.
then be modified to fit each post. Hav-
●● Accuracy and completeness. Check that all your dates tally; otherwise, you
ing a prepared CV on file will reduce
will seem careless. It is better to be honest about your grades and (for exam-
the work each time you apply, while
ple) a period of unemployment than to cover this up or omit details that an
modifying this will help you focus on
employer will want to know. They may be suspicious if you leave things out.
relevant skills and attributes for the par-
ticular job.
Adjusting your CV
You should fine-tune your CV for each submission. Employers frequently use a
Finding vacancies There are quite ‘person specification’ to define the skills and qualities demanded in a job, often
a few different ways that students can under headings such as ‘essential’ and ‘desirable’. This will help you decide
find placements. Some common meth- whether to apply for a position and it assists the selection panel to filter the
ods are: applicants. Highlight relevant qualifications as early in your CV as possible.
●● University placement coordinators
Be selective – don’t include every detail about yourself. Emphasise relevant
liaise with potential hosts and pass parts and leave out irrelevant details, according to the job. Similarly, your letter
knowledge of placements directly of application is not merely a formal document but is also an opportunity for
onto students, persuasion (Box 65.1). You can use it to state your ambitions and highlight par-
●● Vacancies advertised with clear job ticular qualifications and experience. However, do not go over the top – always
descriptions on a company’s own keep the letter to a single page.
website;
●● Vacancies advertised with clear
job descriptions on a university’s
website; Key Point A well constructed and relevant CV will not neces-
●● Students find placements them- sarily guarantee you a job, but it may well get you onto the short
selves; this could be through exist- list for interview. A poor quality CV is a sure route to failure.
ing relationships and contacts, or
through contacting companies they
identify themselves.
Enhancing your CV
Industrial placements are an excellent way to enhance your CV. Placements
have many benefits including:
Industrial placements – provide an
employer a valuable insight to assess
●● Development of your employment-related and employability skills
student competency. This may lead to
(Chapter 58);
future employment with that company. ●● Improved technical and transferable skills (Chapter 58) as well as experience
that adds context to your undergraduate course; and finally,
●● Improving your future career awareness.
A wide variety of chemistry-associated industries (e.g. pharmaceutical, envi-
ronmental, polymer) ranging from large multi-national companies to small –
medium enterprises (SMEs) offer industrial placement to undergraduate
students. Typical industrial placement structures include:
Undergraduate student awareness – ●● a single research project;
your university may provide some
training to assist students with applying
●● a coupled of smaller projects;
for an industrial placement. This may ●● a combination of a research-related project and other related work as might
be via the Industrial Placement Tutor or be undertaken by a graduate chemist; and,
Careers Service.
●● a technical role that might typically be undertaken by a graduate chemist.

Key Point Managing your time (Chapter 59) and working with
others (Chapter 60) are key aspects of an industrial placement –
and your future career in chemical sciences.
Most university chemistry departments will have a designated industrial

University placement tutors – they
placement tutor who is responsible for coordinating links with potential
approve the placement vacancies that
employers, advertising the opportunities as well as offering advice to
their students undertake to ensure the
potential applicants, industrial placements are offered by employers on a
suitability of the work is appropriate.
competitive basis; this will often involve completing an application form.
Selected applicants are then invited to visit the prospective employer for
an interview and a tour of facilities, industrial placements normally take
place in your penultimate year of your degree programme and can occupy
a minimum of 9 months.
During the industrial placement an academic tutor will visit you on at
least one occasion. During this visit to the company the academic tutor will
normally expect to:
●● listen to a short presentation by the placement student (in the presence of the
Placement assessment – the academic work-based supervisor);
tutor will complete an assessment form
post-visit. This is part of the process to ●● hold a private meeting with the placement student;
confirm the suitability of the placement, ●● hold a private meeting with the work-based supervisor; and,
that all parties are happy with progress,
and that no problems have arisen. ●● have a short tour of the workplace environment.
As part of the industrial placement year the student may have to additionally
Placement student – in some cases the undertake, through distance learning, some academic work that contributes to
student may need to undertake some the overall degree classification; this might contribute to the academic process
distance learning during the placement that will ultimately lead to you being awarded a degree with the additional
year to satisfy regulations relating to phrase of ‘with industrial placement’, ‘sandwich’, ‘internship’ or a similar set
the award of their degree. of wording.
The placement student will gain maximum experience and expertise if the
following aspects are in place:
●● You know exactly what is expected of you during the placement. This might
include:
●● understanding your role, that of your work-based supervisor and other
A successful placement – requires employees.
good work-based supervision, support ●● understanding your work and activity boundaries.
from the employer and commitment
from the placement student. ●● understanding how your work performance will be assessed.
●● understanding your contractual working hours (and any expectations
relating to overtime).
Non-disclosure Agreement / Intellectual

●● understanding your role in relation to Good Laboratory Practice and
Property Protection Agreement – this
Health and Safety (Chapter 2).
may need to be signed by both the ●● You are provided with an agreed statement of expectations from the employer
placement student and their academic that includes your duties and responsibilities as well as health and safety.
supervisor (on behalf of the university)
to allow discussion of the placement
●● You are provided with an employment contract that contains specifics on
activities.
terms and conditions of employment, an induction programme and some
relevant training.

Depending upon the employer you may also be provided with a non-disclosure
or intellectual property agreement that covers the information that the company
wishes to protect. Your university will be aware of such agreements and the
implications for use of any of your material in any industrial placement report
that may be required.

Anon. (2000) How to Write a Curriculum Vitae. University Corfield, R. (2009) Preparing the Perfect Job Application.
of London Careers Service, London. Kogan Page, London.
Anon. Graduate Prospects Website. Available: http://www Jackson, A. and Geckeis, K. (2003) How to Prepare your
.prospects.ac.uk Curriculum Vitae. McGraw-Hill, Chicago.
[UK Higher Education Careers Services, containing
good examples of CVs and cover letters]
Study exercises
65.1 Evaluate your personal attributes. Using Print out a copy for filing. Periodically update
Table 65.1, list five qualities that you would use to the word-processed version. If appropriate, save
best describe yourself, and cite the evidence you different versions to be used in different contexts
might give to a potential employer to convince (e.g. when applying for a vacation job).
them that this was the case. List five attributes
65.3 Think about your future career and ask for
you could develop, then indicate how you might
advice. Make an appointment with one of the
do this.
advisers in your university’s careers service. Ask
65.2 Create a generic CV. Drawing on your school about career options for graduates with your
record of achievement, or any CV already pre- intended degree, or determine what qualifica-
pared, e.g. for a part-time job, create a word-pro- tions or module options might be appropriate
cessed generic CV. Save the file in an appropriate for occupations that interest you.
(computer) folder and make a back-up copy.

Communicating information
66. Organising a poster display 575
67. Giving a spoken presentation 581
68. General aspects of scientific writing 587
69. Writing essays 594
70. Reporting practical and project work 597
71. Writing literature surveys and reviews 606

66 Organising a poster display
A scientific poster is a visual display of the results of an investigation, usually

mounted on a rectangular board. Posters are used in undergraduate courses to
display project results or assignment work, and at scientific meetings to com-
municate research findings.
Learning from others – look at the
In a written report you can include a reasonable amount of specific detail
various types of posters around your
and the reader can go back and reread difficult passages. However, if a poster is
university and elsewhere; the best
long-winded or contains too much detail, your reader is likely to lose interest.
examples will be visual, not textual,
with a clear structure that helps get the
key messages across. Key Point A poster session is like a competition – you are
competing for the attention of people in a room. Because you
need to attract and hold the attention of your audience, make
your poster as interesting as possible. Think of it as an advertise-
ment for your work and you will not go far wrong.
Title
1 2 3 4
Preliminaries
(a)
Before considering the content of your poster, you should find out:
●● the linear dimensions of your poster area, typically up to 1.5 m wide by
5 6 7 8
1.0 m high;
●● the composition of the poster board and the method of attachment,
whether drawing pins, Velcro tape, or some other form of adhesive; and
Title whether these will be provided – in any case, it is safer to bring your own;
5 ●● the time(s) when the poster should be set up and when you should attend;
3
(b) 1 ●● the room where the poster session will be held.
6
4 Design
2 7
Plan your poster with your audience in mind, as this will dictate the appropriate
level for your presentation. Aim to make your poster accessible to a broad audi-
ence. Since a poster is a visual display, you must pay particular attention to the
Title presentation of information: work that may have taken hours to prepare can be
ruined in a few minutes by the ill-considered arrangement of items (Fig. 66.1).
Begin by making a draft sketch of the major elements of your poster. It is worth
(c) discussing your intended design with someone else, as constructive advice at
the draft stage will save a lot of time and effort when you prepare the final
version (or consult Sources for further study on p. 584).
Layout
One approach is to divide the poster into several smaller areas, perhaps six or
Fig. 66.1 Poster design. (a) An uninspiring
design: sub-units of equal area, reading
eight in all, and prepare each as a separate item on a piece of card. Alterna-
left to right, are not recommended. (b) This tively, you can produce a single large poster on one sheet of paper or card and
design is more interesting and the text will store it inside a protective cardboard tube. However, a single large poster may
be easier to read (column format). (c) An bend and crease, making it difficult to flatten out. In addition, photographs and
alternative approach, with a central focus text attached to the backing sheet may work loose; a large printed poster with
and arrows/tapes to guide the reader. embedded images is an alternative approach.

Organising a poster display
Subdividing your poster means that each smaller area can be prepared
on a separate piece of paper or card, of A4 size or slightly larger, making
transport and storage easier. It also breaks the reading matter up into smaller
pieces, looking less formidable to a potential reader. By using pieces of card
of different colours you can provide emphasis for key aspects, or link text with
figures or photographs.
Presenting a poster at a formal You will need to guide your reader through the poster and headings/
conference – it can be useful to include sub-headings will help with this aspect. It may be appropriate to use either a
your photograph for identification pur- numbering system, with large, clear numbers at the top of each piece of card,
poses, e.g. in the top right-hand corner or a system of arrows (or thin tapes) to link sections within the poster (see
of the poster. Fig. 66.1). Make sure that the relationship is clear and that the arrows or tapes
do not cross.
Title
Your chosen title should be concise (no more than eight words), specific and
interesting, to encourage people to read the poster, e.g. see Fig 66.2. Make the
title large and bold – it should run across the top of your poster, in letters at
least 4 cm high, so that it can be read from the other side of the room. Coloured
spirit-based marker and block capitals drawn with a ruler work well, as long
as your writing is readable and neat (the colour can be used to add emphasis).
Alternatively, you can print out each word in large font, using a word proces-
sor. Details of authors, together with their addresses (if appropriate), should be
given, usually across the top of the poster in somewhat smaller lettering than
the title.
Making up your poster – text and
graphics printed on good-quality paper Text
can be glued directly onto a contrast-
Write in short sentences and avoid verbosity. Keep your poster as visual as
ing mounting card: use photographic
possible and make effective use of the spaces between the blocks of text (see
spray mountant or glue stick rather
Fig. 66.2). Your final text should be double-spaced and should have a minimum
than liquid glue. Trim carefully using a
capital letter height of 8 mm (minimum font size 36 point), preferably greater,
guillotine to give equal margins, paral-
so that the poster can be read at a distance of 1 m. One method of obtaining text
lel with the paper. Photographs should
of the required size is to photo-enlarge standard typescript (using a good-quality
be scanned into an electronic format,
photocopier), or use a high-quality (laser) printer. It is best to avoid continuous
or placed in a window mount to avoid
use of text in capitals, since it slows reading and makes the text less interesting
the tendency for their corners to curl.
to the reader. Also avoid italic, ‘balloon’ or decorative styles of lettering.
Another approach is to trim pages or
photographs to their correct size, then
encapsulate in plastic film: this gives a Key Point Keep text to a minimum – aim to have a maximum
highly professional finish and is easy to of 500 words in your poster.
transport.
Subtitles and headings

These should have a capital letter height of 15–20 mm, and should be restricted
Producing composite material for to two or three words (see Fig. 66.2). They can be produced by word processor,
p osters – PowerPoint is generally photo-enlargement, by stencilling or by hand, using pencilled guidelines (but
more useful than Word when you wish make sure that no pencil marks are visible on your finished poster).
to include text, graphics and/or images
on the same page. It is possible to use Colour
PowerPoint to produce a complete
poster (Box 66.1), although it can be
Consider the overall visual effect of your chosen display, including the relation-
expensive to have this printed out com-
ship between your text, diagrams and the backing board. Colour can be used to
mercially to A1 or A0 size.
highlight key aspects of your poster (see Fig. 66.2). However, it is very easy to
ruin a poster by the inappropriate choice and application of colour. Careful use
576 Communicating information

Volume 1, Issue 2
2014 BULLETIN
Analysis of volatile compounds (VCs) in
Complex Samples
Chamila J. Denawaka, lan A. Fowlis and John R. Dean
Department of Applied Sciences, Northumbria University, Ellison Building. Newcastle upon Tyne, NE1 8ST, UK.
Aims: An evaluation of static headspace - multicapillary column - gas chromatography - ion mobility spectrometry (SHS-MCC-GC-IMS) has
been undertaken to assess its applicability for the determination of 32 volatile compounds (VCs). The technique was applied to sock malodour
in a field trial using mixed gender volunteers over a 3 week period.
Methods and Results: A FlavourSpec SHS-MCC-GC-IMS instrument (G.A.S. mbH, Dortmund, Germany) was used. A multicapillary column
(MCC) (Multichrom, Novosibirsk, Russia) was used for the chromatographic separation (i.e. 20 cm × 3 mm ID, containing approximately 1000
parallel capillary tubes, 40 mm ID, coated with 0.2 mm film thickness of stationary phase i.e. OV-5 or Carbowax 20M). Atmospheric pressure
ionisation is generated by a Tritium (3H) source. The IMS has a drift tube length of 50 mm. Separation in the IMS drift tube is achieved by applying
an electric field of 2 kV to the ionised volatiles in a pulsed mode using an electronic shutter opening time of 100 ms. The drift gas was N2 (99.998%).
All data are acquired in the positive ion mode and each spectrum is formed with the average of 42 scans.
4
12
xx 1 2 3
7 Gas Drift
out gas in
5
6
8 Carrier gas in
9
11
10
(1) ionisation region; (2) drift region; (3) detector (Faraday plate); (4) radiation
source; (5) multi-capillary column; (6) injector; (7) robotic arm; (8) syringe;
(9) incubator; (10) sample vial (11) integral display; (12) PC. 500
450
400
Retention time (s)
a b 350
1.6
3 300
1.4 xxxxxxx
1.2 2.5 xxxxx 250
Intensity (mV)
Intensity (mV)
1 2 200
0.8 150
0.6 1.5
100
0.4 1
0.2 50
0 0.5 0
-0.2 0 5 6 7 8 9 10 11 12 13 14 15 16
0 500 1000 1500 2000 0 1000 2000 Drift time (ms)
Mass (mg) Mass (mg)
1-butanethiol; ethylbutanoate; 4-fluoroacetophenone (E,E)-2,4-nonadienal
(a) Individual calibration curves of monomer, dimer and trimer and
(b) sum of intensities of monomer, dimer and trimer for 2-phenylethanol. 2-nonanone; (E)-2-nonenal octanal; E-2-octenal; 2-undecanone
HSP
Conclusions: SHS-MCC-GC-IMS has been fully evaluated for the identification of VCs. Based on the optimisation of the key operating
parameters a database. By applying the developed database four VCs were identified and quantified in socks i.e. ammonia, dimethyl
disulphide, dimethyl trisulphide and butyric acid. A link was identified between the presence of high ammonia and dimethyl disulfide
concentrations and a high malodour odour grading i.e. $6. Further work is required to extend the current database and apply the
approach to other sample types and matrices.
Significance and Impact of the Study: SHS-MCC-GC-IMS has been applied to a range
of inorganic and organic volatile compounds. The ease and simplicity of sample preparation,
coupled with the long-term stability of the instrument, make it suitable for a range of studies
requiring sensitive detection of VCs.
Fig. 66.2 An example poster.
of two, or at most three, complementary colours and shades will be easier on

the eye and should aid comprehension. Colour can be used to link the text with
the visual images (e.g. by picking out a colour in a photograph and using the
same colour on the mounting board for the accompanying text). For PowerPoint
posters, careful choice of colours for the various elements will enhance the final
product (Box 66.1). Use coloured inks or water-based paints to provide colour
in diagrams and figures, as felt pens rarely give satisfactory results.

Content
Presenting at a scientific meeting –
never be tempted to spend the mini- The typical format is that of a scientific report (see Box 70.1), i.e. with the same
mum amount of time converting a piece headings, but with a considerably reduced content. Keep references within the
of scientific writing into poster format – text to a minimum – interested parties can always ask you for further informa-
the least interesting posters are those tion. Also note that most posters have a summary/conclusions section at the
where the author simply displays pages end (see Fig. 66.2), rather than an abstract.
from a written communication (e.g. a
journal article) on a poster board.
Introduction
This should give the reader background information on the broad field of study
and the aims of your own work. It is vital that this section is as interesting as
possible, to capture the interest of your audience. It is often worth listing your
Designing the materials and methods
objectives as a series of numbered points.
section – photographs or diagrams of
apparatus can help to break up the text
of this section and provide visual inter- Experimental
est. It is sometimes worth preparing Keep this short, and describe only the principal techniques used (see Fig. 66.2).
this section in a smaller typeface. You might mention any special techniques, or problems of general interest.
Results
Do not present your raw data: use data reduction wherever possible, i.e. figures
Keeping graphs and diagrams simple –
and simple statistical comparisons (see Fig. 66.2). Graphs, diagrams, histo-
avoid composite graphs with different
grams and pie charts give clear visual images of trends and relationships and
scales for the same axis, or with several
should be used in place of data tables (see p. 451). Final copies of all figures
trend lines (use a maximum of three
should be produced so that the numbers can be read from a distance of 1 m.
trend lines per graph).
Each should have a concise title and legend, so that it is self-contained: if
appropriate, a series of numbered points can be used to link a diagram with
the accompanying text. Where symbols are used, provide a key on each graph
(symbol size should be at least 5 mm). Avoid using graphs straight from a
written version, e.g. a project report, textbook or a paper, without considering
whether they need modification to meet your requirements.
Conclusions
This is where many readers will begin, and they may go no further unless you
Listing your conclusions – a series of
make this section sufficiently interesting. This part needs to be the strongest
numbered points is a useful approach,
section of your poster, summarising the main points (see Fig. 66.2). Refer to
if your findings fit this pattern.
your figures here to draw the reader into the main part of your poster. A slightly
larger or bolder typeface may add emphasis, though too many different type-
faces can look messy. For the reference list, a smaller font can be used.
The poster session

Consider providing a handout – this A poster display session may be organised as part of the assessment of your
is a useful way to summarise the main coursework, and this usually means those held at scientific meetings and con-
points of your poster, so that your read- ferences. Staff and fellow students (delegates at conferences) will mill around,
ers have a permanent record of the looking at the posters and chatting to their authors, who are usually expected
information you have presented. to be in attendance. If you stand at the side of your poster throughout the ses-
sion you are likely to discourage some readers, who may not wish to become
involved in a detailed conversation about the poster. Stand nearby. Find some-
thing to do – talk to someone else, or browse among the other posters, but
remain aware of people reading your poster and be ready to answer any queries

Box 66.1 How to create a poster using Microsoft PowerPoint
Software such as PowerPoint can be used to produce a have your final images, use blank text boxes to show
high-quality poster, providing you have access to a good their position within the poster.
colour printer. However, you should avoid the standard
5. Add text. Use either the Insert tab to select a Text
templates available on the web as they encourage unnec-
Box and place this on your slide, then either type
essary uniformity and stifle creativity, leading to a less
in your text (use the Enter key to provide line spac-
satisfying end result. The following steps give practical
ing within the box) or copy-and-paste text from a
advice on creating a poster as a single PowerPoint slide:
word-processed file. You will need to consider the
1. Sketch out your plans. Decide on the main poster ele- font size for the printed poster (e.g. for an A0 poster
ments (images, graphs, tables and text sections) and (size 1189 * 841 mm), a printed font size of 24 point
their relationship with each other and draw out a one- is appropriate for the main text, with larger fonts for
page ‘storyboard’ (see Fig. 66.1).Think about colours for headings and titles. If you find things difficult to read
background, text and graphics (use two or three com- on-screen, use the Zoom function on the View tab
plementary colours) – dark text on a light background (either select a larger percentage in the Zoom box on
is clearer (high contrast), and uses less ink when print- the bottom right toolbar, or hold down the Control
ing. Also consider how you will link the elements in key and use the mouse wheel to scroll up (magnify)
sequence, to guide readers through your ‘story’. or down (reduce). Use a separate text box for each
element of your poster and do not be tempted to type
2. Get your material ready. Collect together individ- too much text into each box – write in phrases, using
ual files for pictures, figures and tables. Make any bullet points and numbered lists to keep text concise
required adjustments to images, graphs or tables (aim for no more than 50 words per text box). Select
before you import them into your poster. appropriate font styles and colours using the Home
3. Create a new/blank slide. Open PowerPoint From the tab, Font menu. For a background colour or surround-
Home tab, select New Slide and then Blank. Then use ing line, right-click and use the Format Text Effects box
the Design tab 7 Page setup group to select either Format Shape box commands (line thickness and col-
Landscape or Portrait orientation and to set the cor- our can then be altered). Present supplementary text
rect page size (use Width and Height commands, or elements in a smaller font – for example, details of
select a standard size like A4, A3, A2, etc.). Right-click methodology, references cited.
on the slide and select Ruler and Grid and Guides (to 6. Add boxes, lines and/or arrows to link elements of the
help position elements within the slide – the horizon- poster and guide the reader (see Fig. 66.1). These fea-
tal and vertical guidelines can be dragged to different tures are available from the Home tab. Drawing group
positions at later stages, as required). Use the tem- drop-down menu. Note that new inserts are overlaid
plates (Themes) on the top line menu or select an over older inserts – if this proves to be a problem, select
appropriate background style and colour. In general, the relevant item and use the Arrange, Order functions
avoid setting a picture as your background as these within the Drawing group to change its relative position.
tend to detract from the content of the poster. Before
going further, save your work. Repeat this frequently 7. Review your poster. Get feedback from another stu-
and in more than one location (e.g. hard drive and dent or tutor, e.g. on a small printed version, or use a
USB memory stick). projector to view your poster without printing (adjust
the distance between projector and screen to give the
4. Add graphics. For images, use the Insert tab, select correct size).
Picture and browse to Insert the correct file. The Insert,
Object command performs a similar function for Excel 8. Revise and edit your poster. Revisit your work and
charts (graphs). Alternatively, use the copy-and-paste remove as much text as possible. Delete any compo-
functions of complementary software. Once inserted, nent that is not essential to the message of the poster.
resize using the sizing handles in one of the corners Keep graphs simple and clear (p. 451 gives further
(for photographs, take care not to alter one dimension advice). White space is important in providing structure.
relative to the other, or the image will be distorted).
9. Print the final version. Use a high-resolution colour
To re-position, put the mouse pointer over the image,
printer (this may be costly, so you should wait until
left-click and hold, then drag to new location. While
you are sure that no further changes are needed).
the Insert tab offers standard shapes and other useful
features, you should avoid clipart (jaded and over- Note: PowerPoint commands illustrated here may very
used) and poor-quality images from the web (always among the different versions of this Microsoft Office
use the highest resolution possible) – if you do not program.

they may raise. Do not be too discouraged if you are not asked lots of questions:
Coping with questions in assessed
remember, the poster is meant to be a self-contained, visual story, without need
poster sessions – you should expect to
for further explanation.
be asked questions about your poster,
A poster display will never feel like an oral presentation, where the nerv-
and to explain details of figures, meth-
ousness beforehand is replaced by a combination of satisfaction and relief as
ods, etc.
you unwind after the event. However, it can be a very satisfying means of
communication, particularly if you follow these guidelines.

Alley, M. (2007) The Craft of Scientific Presentations: Crit- Gosling, P.J. (1999) Scientist’s Guide to Poster Presenta-
ical Steps to Succeed and Critical Errors to Avoid, 5th tions. Kluwer, New York.
edn. Springer-Verlag, New York. Hess, G., Tosney, K. and Liegel, L. Creating Effective
Briscoe, M.H. (1996) Preparing Scientific Illustrations: A Poster Presentations. Available: http://www.ncsu.edu/
Guide to Better Posters, Presentations and Publications, project/posters/.
2nd edn. Springer-Verlag, New York. Last accessed 01/09/15.
Davis, M.F. (2004) Scientific Papers and Presentations, 2nd Nicol, A.A.M. (2010) Displaying Your Findings, 6th edn.
edn. Academic Press, New York. American Psychological Association, Washington.
Study exercises
66.1 Design a poster. Working with one or more part- good poster presentation?’ Compare your list with
ners from your year group, decide on a suitable the one that we have provided online – do you
poster topic (perhaps something linked to your agree with our criteria, or do you prefer your own
current teaching programme). Working individu- list (and can you justify your preferences)?
ally, make an outline plan of the major elements
66.3 Evaluate the posters in your university. Most
of the poster, with appropriate sub-headings and
universities have a wide range of academic post-
a brief indication of the content and relative size
ers on display. Some may cover general topics
of each element (including figures, diagrams and
(e.g. course structures), while others may deal
images). Exchange draft plans with your partners
with specific research topics (e.g. poster pres-
and arrange a session where you can discuss
entations from past conferences). Consider their
their merits and disadvantages.
good and bad features (if you wish to make this
66.2 Prepare a checklist for assessing the quality of a a group exercise, you might compare your eval-
poster presentation. After reading through this uation with that of other students, in a group dis-
chapter, prepare a ten-point checklist of assess- cussion session).
ment criteria under the heading ‘What makes a

67 Giving a spoken presentation
Most students feel very nervous about giving talks. This is natural, since very
Opportunities for practising speaking
few people are sufficiently confident and outgoing that they look forward to
skills – these include:
speaking in public. Additionally, the technical nature of the subject matter may
●● answering lecturers’ questions; give you cause for concern, especially if you feel that some members of the
●● contributing in tutorials; audience have a greater knowledge than you have, e.g. your tutors. However,
●● talking to informal groups; this is a fundamental method of scientific communication and an important
●● giving your views at formal transferable skill; therefore, it forms an important component of many courses.
(committee) meetings; The comments in this chapter apply equally to informal talks, e.g. those
●● demonstrating or explaining to other
based on assignments and project work, and to more formal conference pres-
students, e.g. during a practical class;
●● asking questions in lectures/seminars;
entations. It is hoped that the advice and guidance given below will encourage
●● answering an examiner’s questions you to make the most of your opportunities for public speaking, but there is no
in an oral exam. substitute for practice. Do not expect to find all of the answers from this, or any
other, book. Rehearse, and learn from your own experience.
Key Point The three ‘Rs’ of successful public speaking are:

reflect – give sufficient thought to all aspects of your presenta-
tion, particularly at the planning stage; rehearse – to improve
your delivery; revise – modify the content and style of your
material in response to your own ideas and to the comments
of others.
Preparation
Learning from experience – use your
own experience of good and bad lectur- Preliminary information
ers to shape your performance. Some Begin by marshalling the details needed to plan your presentation, including:
of the more common errors include:
●● the duration of the talk;
●● speaking too quickly;
●● reading from notes or from slides ●● whether time for questions is included;
and ignoring the audience;
●● inexpressive, impersonal or indistinct ●● the size and location of the room;
speech; ●● the projection/lighting facilities provided, and whether pointers or similar
●● distracting mannerisms; aids are available.
●● poorly structured material with little
emphasis on key information; It is especially important to find out whether the room has the necessary equip-
●● factual information too complex and ment for digital projection (e.g. PC, projector and screen, black-out curtains
detailed; or blinds, appropriate lighting) or overhead projection before you prepare your
●● too few or too many visual aids. audio-visual aids. If you concentrate only on the spoken part of your presenta-
tion at this stage, you are inviting trouble later on. Have a look around the room
and try out the equipment at the earliest opportunity, so that you are able to
use the lights, projector, etc. with confidence. For digital projection systems,
Testing the room – if possible, try to check that you can load/present your material. Box 67.1 gives practical advice
rehearse your talk in the room in which on the use of PowerPoint.
it will be presented. This will help you
to make allowance for layout of equip- Audio-visual aids
ment, lighting, acoustics and sight lines If you plan to use overhead transparencies, find out whether your department
that might affect the way you deliver has facilities for their preparation, whether these facilities are available for your
your talk. It will also put you more at use, and the cost of materials. Adopt the following guidelines:
ease on the day, because of the famili-
arity of the surroundings.
●● Keep text to a minimum: present only the key points, with up to 20 words
per slide/transparency.

Giving a spoken presentation
Box 67.1 Tips on preparing and using Microsoft PowerPoint slides in a spoken presentation
Microsoft PowerPoint can be used to produce high- type whole sentences as you will then be reduced to
quality visual aids, assuming a computer and digital reading these from the screen during your presenta-
projector are available in the room where you intend tion, which is boring.
to speak. The presentation is produced as a series of
●● Use a large, clear font style. Use the Slide Master
electronic ‘slides’ onto which you can insert images,
option within the View menu to set the default font
diagrams and text. When creating your slides, bear the
to a non-serif style such as Arial, or Comic Sans
following points in mind:
MS, and an appropriate colour. Default fonts for head-
●● Plan the structure of your presentation. Decide on the ings and bullet points are intentionally large, for clar-
main topic areas and sketch out your ideas on paper. ity. Do not reduce these to anything less than 28 point
Think about what material you will need (e.g. pictures, font size (preferably larger) to cram in more words: if
graphs) and what colours to use for background and you have too much material, create a new slide and
text. divide up the information.
●● Choose slide layouts according to purpose. Once ●● Animate your material. The Animation tab provides a
PowerPoint is running, from the Home tab select Lay- function that enables you to introduce the various ele-
out. You can then add material to each new slide to suit ments within a slide, e.g. text can be made to Appear
your requirements. one line at a time, to prevent the audience from read-
ing ahead and help maintain their attention.
●● Select your background with care. Many of the preset
background templates (Themes) available within the ●● Do not overdo the special effects. PowerPoint has a
Design tab are best avoided, since they are over-used wide range of features that allow complex slide transi-
and fussy, diverting attention from the content of the tions and animations, additional sounds, etc. but these
slides. Conversely, flat, dull backgrounds may seem quickly become irritating to an audience unless they
uninteresting, while brightly coloured backgrounds have a specific purpose within your presentation.
can be garish and distracting. Choose whether to ●● Always edit your slides before use. Check through
present your text as a light-coloured font on a dark your slides and cut out any unnecessary words, adjust
background (more restful but perhaps less engaging the layout and animation. Remember the maxim ‘less
if the room is dark) or a dark-coloured font on a light is more’ – avoid too much text; too many bullet points;
background (more lively). too many distracting visual effects or sounds.
●● Use visual images throughout. Remember the maxim
When presenting your talk:
‘a picture is worth ten thousand words’. A presenta-
tion composed entirely of text-based slides will be ●● Work out the basic procedures beforehand. Practise, to
uninteresting: adding images and diagrams will make sure that you know how to move forwards and
brighten up your talk considerably (use the Insert backwards in your slideshow, turn the screen on and
menu, Picture option). Images can be taken with a off, hide the mouse pointer, etc.
digital camera, scanned in from a printed version or
●● Do not forget to engage your audience. Despite the
copied and pasted from the web, but you should take
technical gadgetry, you need to play an active role
care not to break copyright regulations. ‘Clipart’ is
in the presentation, as explained elsewhere in this
copyright-free, but should be used sparingly as most
chapter.
people will have seen the images before and they are
rarely wholly relevant. Diagrams can be made from ●● Do not go too fast. Sometimes, new users tend to
components created using the Home tab Drawing deliver their material too quickly: try to speak at a nor-
group while graphs and tables can be imported from mal pace and practise beforehand.
other programs, e.g. Excel (Box 15.1 gives further ●● Consider whether to provide a handout. PowerPoint
specific practical advice on adding graphics, saving has several options, including some that provide
files, etc.). space for notes (e.g. Fig. 61.3). However, a handout
●● Keep text to a minimum. Aim for no more than should not be your default option, as there is a cost
20 words on a single slide (e.g. four/five lines con- involved.
taining a few words per line). Use headings and sub-
Note: PowerPoint commands illustrated here very
headings to structure your talk: write only key words
among the different versions of the Microsoft Office
or phrases as ‘prompts’ to remind you to cover a par-
program.
ticular point during your talk – never be tempted to

●● Make sure the text is readable: try out your material beforehand.
●● Use several simpler figures rather than a single complex graph.
Using audio-visual aids – do not let
●● Avoid too much colour on slides: blue and black are easier to read than
the equipment and computer gadgetry
red or green.
distract you from the essential rules of
good speaking (Box 67.2): remember ●● Do not mix slides and transparencies as this is often distracting.
that you are the presenter.
●● Use spirit-based pens for transparencies: use alcohol for corrections.
●● Transparencies can be produced from typewritten or printed text using
a photocopier, often giving a better product than pens. Note that you must
use special heat-resistant acetate sheets for photocopying.
Electronic presentation software (e.g. PowerPoint) can replace these specialist
requirements, as long as the necessary facilities are available for your talk (see
below).
Audience
You should consider your audience at the earliest stage, since they will deter-
Pitching your talk at the right level –
mine the appropriate level for your presentation. If you are talking to fellow
the general rule should be: ‘do not over-
students you may be able to assume a common level of background knowl-
estimate the background knowledge of
edge. In contrast, a research lecture given to your department, or a paper
your audience’. This sometimes hap-
at a meeting of a scientific society, will be presented to an audience from
pens in student presentations, where
a broader range of backgrounds. An oral presentation is not the place for
fears about the presence of ‘experts’
a complex discussion of specialised information: build up your talk from a
can encourage the speaker to include
low level. The speed at which this can be done will vary according to your
too much detail, overloading the audi-
audience. As long as you are not boring or patronising, you can cover basic
ence with facts.
information without losing the attention of the more knowledgeable members
in your audience.
Content
While the specific details in your talk will be for you to decide, most spoken
presentations share some common features of structure, as described below.
Getting the introduction right –

a good idea is to have an initial slide Introductory remarks
giving your details and the title of your It is vital to capture the attention of your audience at the outset. Consequently,
talk, and a second slide telling the you must make sure your opening comments are strong, otherwise your audi-
audience how your presentation will ence will lose interest before you reach the main message. Remember it takes a
be structured. Make eye contact with sentence or two for an audience to establish a relationship with a new speaker.
all sections of the audience during the Your opening sentence should be some form of preamble and should not con-
introduction. tain any key information. For a formal lecture, you might begin with ‘Thank
you for that introduction. My talk today is about . . . ’ then restate the title and
acknowledge other contributors, etc. You might show a transparency or slide
with the title printed on it, or an introductory photograph, if appropriate. This
What to cover in your introduction – should provide the necessary settling-in period.
You should: After these preliminaries, you should introduce your topic. Begin your
●● explain the structure of your talk; story on a strong note – avoid timid or apologetic phrases.
●● set out your aims and objectives; Opening remarks are unlikely to occupy more than 10% of the talk. How-
●● explain your approach to the topic. ever, because of their significance, you might reasonably spend up to 25% of
your preparation time on them.

Key Point Make sure you have practised your opening

remarks, so that you can deliver the material in a flowing style,
with less chance of mistakes.
The main message

Allowing time for slides – as a rough
guide you should allow at least two This section should include the bulk of your experimental results or literature
minutes per illustration, although some findings, depending on the type of presentation. Keep details of methods to
diagrams may need longer, depending the minimum needed to explain your data. This is not the place for a detailed
on content. Make a note of the halfway description of equipment and experimental protocol (unless it is a talk about
point, to help you check timing/pace. methodology). Results should be presented in an easily digested format.
Key Point Do not expect your audience to cope with large

amounts of data; use a maximum of six numbers per slide.
Remember that graphs and diagrams are usually better than
tables of raw data, since the audience will be able to see the
visual trends and relationships in your data (p. 451).
Present summary statistics (Chapter 53) rather than individual results.

Show the final results of any analyses in terms of the statistics calculated, and
their significance (p. 495), rather than dwelling on details of the procedures
concluding introduction used. Figures should not be crowded with unnecessary detail. Every diagram
remarks (10%) (10%) should have a concise title and the symbols and trend lines should be clearly
labelled, with an explanatory key where necessary. When presenting graphical
data (Chapter 49) always ‘introduce’ each graph by stating the units for each
axis and describing the relationship for each trend line or data set.
main message Key Point Use summary slides at regular intervals, to maintain
(80%)
the flow of the presentation and to emphasise the main points.
Take the audience through your story step-by-step at a reasonable pace. Try
not to rush the delivery of your main message due to nervousness. Avoid com-
note halfway point plex, convoluted story-lines – one of the most distracting things you can do is to
fumble backwards through PowerPoint slides or overhead transparencies. If you
Fig. 67.1 Pie chart showing time allocation
need to use the same diagram or graph more than once then you should make
for a typical presentation.
two (or more) copies. In a presentation of experimental results, you should
discuss each point as it is raised, in contrast to written text where the results and
discussion may be in separate sections. The main message typically occupies
approximately 80% of the time allocated to an oral presentation (Fig. 67.1).
Final remarks – make sure you give the Concluding remarks

audience sufficient time to assimilate
your final slide: some of them may wish
Having captured the interest of your audience in the introduction and given
to write down the key points. Alterna-
them the details of your story in the middle section, you must now bring your
tively, you might provide a handout,
talk to a conclusion. Do not end weakly, e.g. by running out of steam on the last
with a brief outline of the aims of your
slide. Provide your audience with a clear ‘take-home message’, by returning
study and the major conclusions.
to the key points in your presentation. It is often appropriate to prepare a slide
or overhead transparency listing your main conclusions as a numbered series.
Signal the end of your talk by saying ‘finally . . . ’, ‘in conclusion . . . ’, or
a similar comment and then finish speaking after that sentence. Your audience
will lose interest if you extend your closing remarks beyond this point. You

may add a simple end phrase (for example, ‘thank you’) as you put your notes
into your folder, but do not say ‘that’s all folks!’, or make any similar offhand
remark. Finish as strongly and as clearly as you started. Box 67.2 gives further
advice on presentation.
Box 67.2 Hints on spoken presentations
In planning the delivery of your talk, bear the following notes. These timing marks will help you keep to
aspects in mind: time during the final presentation.
●● Using notes. Many accomplished speakers use abbre- (c) Avoide looking at your wristwatch when speak-
viated notes for guidance, rather than reading wording: this sends a negative signal to the audience.
for-word from a prepared script. When writing your talk: Use a wall clock (where available), or take off your
watch and put it beside your notes so that you
(a) Consider Preparing your first draft as a full script: can glance at it without distracting the audience.
write in spoken English and keep the text simple, to
avoid a formal, impersonal style. Your Aim should ●● Consider your image. Make sure that the image you
be to talk to your audience, not read to them. project is appropriate for the occasion:
(b) If necessary, use note-cards with key words and (a) Think about what to wear: aim to be respectable
phrases: it is best to avoid using a full script in without ’dressing up’, otherwise your message
the final presentation. As you rehearse and your may be diminished.
confidence improves, a set of note-cards may be (b) Maintain a good posture: it will help your voice
an more appropriate format. Mark the position projection if you stand upright, rather than
of slides/key points, etc,: each note-card should slouching or leaning over the lectern.
contain details of structure as well as content.
(c) Deliver your material with expression: Project
Your notes should be written/printed in text large
your voice towards the audience at the back of
enough to be read easily during the presentation
the room and make sure you look round to make
(also check that the lecture room has a lectern
eye contact with all sections of the audience.
light or you may have problems reading your
Arm movements and subdued body language
notes if the lights are dimmed). Each note-card
will help maintain the interest of your audience.
or sheet should be clearly numbered, so that you
However, you should avoid extreme gestures (it
do not lose your place.
may work for some TV personalities but it is not
(c) Decide on the layout of your talk: give each recommended for the beginner).
sub-division a heading in your notes, so that your
(d) Try to identify and control any repetitive manner-
audience is made aware of the structure.
isms: repeated ’empty’ words/phrases, fidgeting
(d) Memorise your introductory/closing remarks: with pens, keys, etc.) will distract your audience.
you may prefer to raly on a full written version Note-cards held in your hand give you something
for these sections, in case your memory fails, or to focus on, while laser pointers will show up any
if you suffer ’stage fright’. nervous hand tremors. Practising in front of a
(e) Using PowerPoint (Box 67.1): here, you can either mirror may help.
use the ’notes’ option (View tab; Notes Page), ●● Think about questions. Once again, the best approach
or you may even prefer to dispense with notes is to prepare beforehand:
entirely, since the slides will help structure your
talk, acting as an aide-memoire for your material. (a) Consider what questions are likely to come up,
and prepare brief answers. However, do not be
●● Work on your timing. It is essential that your talk is the
afraid to say ’I don’t know’: your audience will
right lenght and the correct pace:
appreciate honesty rather than vacillation if you
(a) Rehearse your presentation: ask a friend to lis- do not have an answer for a particular question.
ten and to comment constructively on those (b) If no questions are asked, you might pose a ques-
parts that were difficult to follow, to improve your tion yourself and then ask for opinions from the
performance. audience: if you use this approach, you should be
(b) Use ’split times’ to pace yourself: following an prepare to comment briefly if your audience has
initial run-through, add the times at which you no suggestions, to avoid the presentation ending
should arrive at the key points of your talk to your in an embarrrassing silence.


Alley, M. (2007) The Craft of Scientific Presentations: Kranz, W. B. (2016) Oral Communication Strategies for
Critical Steps to Succeed and Critical Errors to Avoid. Scientific Presentation. Academic Press Inc., New York.
5th edn. Springer-Verlag, New York. Radel, J. Oral Presentations. Available: http://people.eku
Capp, C.C. and Capp, G.R. (1989) Basic Oral Communi- .edu/ritchisong/oralpres.html
cation, 5th edn. Prentice Hall, Harlow. Last accessed 01/09/150.
Study exercises
67.1 Prepare a checklist for assessing the quality of speakers, presenters of TV documentaries, etc.
an oral presentation. After reading through this Decide in advance how you are going to tackle
chapter, prepare a ten-point checklist of assess- the evaluation (e.g. with a quantitative marking
ment criteria under the heading ‘What makes a scheme, or a less formal procedure).
good oral presentation?’. Compare your list with
67.3 Rehearse a talk and get feedback on your perfor-
the one that we have provided online – do you
mance. There are a number of approaches you
agree with our criteria, or do you prefer your
might take, including: (i) recording and reviewing
checklist? (Can you justify your preferences?)
your presentation using a digital movie camera;
67.2 Evaluate the presentation styles of other speakers. or (ii) giving your talk to a small group of fellow
There are many opportunities to assess the students and asking them to provide constructive
strengths and weaknesses of academic ‘pub- feedback.
lic speakers’, including your lecturers, seminar

68 General aspects of scientific writing
Written communication is an essential component of all sciences. Most courses

include writing exercises in which you will learn to describe ideas and results
accurately, succinctly and in an appropriate style and format. The following
features/aspects are common to all forms of scientific writing.
Time management – practical advice is Organising your time

given in Chapter 59.
Making a timetable at the outset helps ensure that you give each stage adequate
attention and complete the work on time (e.g. Fig. 68.1). To create and use a
timetable:
Monday: morning Lectures (University)
afternoon Practical (University)
1. Break down the task into stages.
evening Initial analysis and 2. Decide on the proportion of the total time each stage should take.
brainstorming (Home)
3. Set realistic deadlines for completing each stage, allowing some time
Tuesday: morning Lectures (University)
afternoon Locate sources (Library)
for slippage.
evening Background reading (Library) 4. Refer to your timetable frequently as you work: if you fail to meet one
Wednesday: morning Background reading (Library) of your deadlines, make a serious effort to catch up as soon as possible.
afternoon Squash (Sports hall)
evening Planning (Home)
Key Point The appropriate allocation of your time to reading,
Thursday: morning Lectures (University)
planning, writing and revising will differ according to the task in
afternoon Additional reading (Library)
evening Prepare outline (Library)
hand (see Chapters 69–71).
Friday: morning Lab class (University)
afternoon Write first draft (Home) Organising your information and ideas
evening Complete first draft (Home)
Before you write, you need to gather and/or think about relevant material
Saturday: morning Shopping (Town)
(Chapters 41 and 42). You must then decide:
afternoon Review first draft (Home)
evening Revise first draft (Home) ●● what needs to be included and what does not;
Sunday: morning Free
●● in what order it should appear.
afternoon Produce final copy (Home)
evening Proof read and print Start by jotting down headings for everything of potential relevance to the
essay (Home) topic (this is sometimes called ‘brainstorming’). A spider diagram (Fig. 68.2)
Monday: morning Final read-through and check or a mind map (Fig. 61.2) will help you organise these ideas. The next stage is
Submit essay to create an outline of your text (Fig. 67.3). Outlines are valuable because they:
(deadline midday)
Go out with friends ●● force you to think about and plan the structure;
Fig. 68.1 Example timetable for writing a

●● provide a checklist so nothing is missed out;
short essay. ●● ensure the material is balanced in content and length;
●● help you organise figures and tables, by showing where they will be used.
Key Point A suitable structure is essential to the narrative of

your writing, and should be carefully considered at the outset.
Creating an outline – an informal out-

line can be made simply by indicating In an essay or review, the structure of your writing should help the reader
the order of sections on a spider dia- to assimilate and understand your main points. Subdivisions of the topic could
gram (as shown in Fig. 68.2). simply be related to the physical nature of the subject matter (e.g. compo-
nents of a gas chromatograph) and should proceed logically (e.g. injection
port, column). A chronological approach is good for evaluation of past work

General aspects of scientific writing
analytes sample is ionised the masses of the

linear range 106 destroyed into fragments fragments are detected
detects organic library of fragment

flame ionisation destructive mass spectrometry
compounds which ionise patterns available
detector detector
in a hydrogen flame for comparison
universal universal linear range 105
Gas Chromatography Detectors
responds to used for analysis used for gas purity base line affected
electronegative of herbicides tests for by changes in
substituents on analytes and PCBs refinery gases flow rate
components must have

electron capture thermal conductivity
non-destructive conductivities different
detector detector
from the carrier gas
linear range 104 selective analytes are linear range 106 universal
not altered
Fig. 68.2 Spider diagram showing how you might ‘brainstorm’ an essay with the title ‘Detectors for Gas Chromatog-
raphy’. Write out the essay title in full to form the spider’s body, and as you think of possible content, place headings
around this to form its legs. Decide which headings are relevant and which are not and use arrows to note connections
between subjects, if required. This may influence your choice of order and may help to make your writing flow because
the links between paragraphs will be natural. You can make an informal outline directly on a spider diagram by adding
numbers indicating a sequence of paragraphs (as shown). This method is best when you must work quickly, as with an
essay written under exam conditions.
Gas chromatography detectors Gas chromatography detectors
Introduction 1. Introduction
Destructive detectors 2. Destructive detectors
Flame ionisation 2.1 Flame ionisation
Mass spectrometry 2.2 Mass spectrometry
Non-destructive detectors 3. Non-destructive detectors
Thermal conductivity 3.1 Thermal conductivity
Electron capture 3.2 Electron capture
Conclusions 4. Conclusions
(a) (b)
Fig. 68.3 Formal outlines. These are useful for a long piece of work where
you or the reader might otherwise lose track of the structure. The headings
for sections and paragraphs are simply written in sequence with the type
of lettering and level of indentation indicating their hierarchy. Two different
forms of formal outline are shown, a minimal form (a) and a numbered form
(b). Note that the headings used in an outline are often repeated within the
essay to emphasise its structure. The content of an outline will depend on
the time you have available and the nature of the work, but the most detailed
hierarchy you should reasonably include is the subject of each paragraph.
(e.g. the development of the concept of aromaticity), whereas a step-by-step

comparison might be best for certain exam questions (e.g. ‘Discuss the differ-
ences between solid phase extraction and Soxhlet extraction in the recovery

of pesticides’). There is little choice about structure for practical and project
reports (see p. 602).
Writing
Adopting a scientific style
Your main aim in developing a scientific style should be to put your message
across directly and unambiguously. While you can try to achieve this through
a set of ‘rules’ (see Box 68.1), you may find other requirements driving your
writing in a contradictory direction. For instance, the need to be accurate
and complete may result in text littered with technical terms, and the flow
may be continually interrupted by references to the literature. The need to
be succinct also affects style and readability through the use of, for example,
stacked noun-adjectives (e.g. ‘ring opening metathesis polymerisation’) and
acronyms (e.g. ‘ROMP’). Finally, style is very much a matter of taste and
each tutor, examiner, supervisor or editor will have pet loves and hates which
you may have to accommodate. Different assignments will need different
styles; Box 68.2 gives further details.
Developing technique
Improving your writing skills – you
Writing is a skill that can be improved, but not instantly. You should analyse
need to take a long-term view if
your deficiencies with the help of feedback from your tutors, be prepared to
you wish to improve this aspect of your
change work habits (e.g. start planning your work more carefully, or find out
work. An essential preliminary is to
more about punctuation), and be willing to learn from some of the excellent
invest in and make full use of a personal
texts that are available on scientific writing (p. 597).
reference library (see Box 68.3).
Getting started
A common problem is ‘writer’s block’ – inactivity or stalling brought on by a
variety of causes. If blocked, ask yourself these questions:
Benefits of talking about your ●● Are you comfortable with your surroundings? Make sure you are seated
work – discussing your topic with a comfortably at a reasonably clear desk and have minimised the possibility
friend or colleague might bring out of interruptions and distractions.
ideas or reveal deficiencies in your
knowledge. ●● Are you trying to write too soon? Have you clarified your thoughts on the
subject? Have you done enough preliminary reading?
●● Are you happy with the underlying structure of your work? If you have
not made an outline, try this. If you are unhappy because you cannot think of
a particular detail at the planning stage, just start writing – it is more likely
to come to you while you are thinking of something else.
Writing with a word processor – use ●● Are you trying to be too clever? Your first sentence does not have to be
the dynamic/interactive features of the earth-shattering in content or particularly smart in style. A short statement
word processor (Chapter 46) to help you of fact or a definition is fine. If there will be time for revision, first get your
get started: first make notes on struc- ideas down on paper and then revise grammar, content and order later.
ture and content, then expand these ●● Do you really need to start writing at the beginning? Try writing the
to form a first draft and finally revise/ opening remarks after a more straightforward part. For example, with reports
improve the text. of laboratory work, the Materials and Methods section may be the easiest
place to start.
●● Are you too tired to work? Do not try to ‘sweat it out’ by writing for long
periods at a stretch: stop frequently for a rest.

Box 68.1 How to achieve a clear, readable style
Words and phrases contractions are generally not used in formal scien-
tific writing.
●● Choose short, clear words and phrases rather than
long ones: e.g. use ‘build’ rather than ‘fabricate’; ‘now’ – Do not use an apostrophe for ‘its’ as the possessive
rather than ‘at the present time’. At certain times, tech- form of ‘it’ (e.g. ‘the university and its surroundings’).
nical terms must be used for precision, but do not use Note that ‘it’s’ is reserved for ‘it is’. This is an excep-
jargon if you do not have to. tion to the general rule and a very common mistake.
●● Do not worry too much about repeating words, espe- – Never use an apostrophe to indicate plurals. Even
cially when introducing an alternative might subtly for abbreviations, the accepted style is now to omit
alter your meaning. the apostrophe for the plural (e.g. write ‘the ELISAs
were . . . ’).
●● Where appropriate, use the first person to describe
your actions (‘We decided to’; ‘I conclude that’), but not Sentences
if this is specifically discouraged by your supervisor or
●● Do not make them overlong or complicated.
department.
●● Introduce variety in structure and length.
●● Favour active forms of writing (‘the observer com-
pleted the survey in ten minutes’) rather than a pas- ●● If unhappy with the structure of a sentence, try chop-
sive style (‘the survey was completed by the observer ping it into a series of shorter sentences.
in ten minutes’).
Paragraphs
●● Use tenses consistently. Past tense is always used for
●● Get the paragraph length right – five sentences or so. Do
materials and methods (‘samples were taken from
not submit an essay that consists of a single paragraph,
. . . ’) and for reviewing past work (‘Smith (1990) con-
nor one that contains single sentence paragraphs.
cluded that . . . ’). The present tense is used when
describing data (‘Fig. 1 shows . . . ’), for generalisations ●● Make sure each paragraph is logical, dealing with a
(‘Most authorities agree that . . . ’) and conclusions (‘To single topic or theme.
conclude, . . . ’).
●● Take care with the first sentence in a paragraph (the
●● Use statements in parentheses sparingly – they disrupt ‘topic’ sentence); this introduces the theme of the
the reader’s attention to your central theme. paragraph. Further sentences should then develop
this theme, e.g. by providing supporting information,
●● Avoid clichés and colloquialisms – they are usually
examples or contrasting cases.
inappropriate in a scientific context.
●● Use ‘linking’ words or phrases to maintain the flow
Punctuation
of the text within a paragraph (e.g. ‘for example’; ‘in
●● Try to use a variety of types of punctuation, to make contrast’; ‘however’; ‘on the other hand’).
the text more interesting to read.
●● Make your text more readable by adopting modern
●● Decide whether you wish to use ‘closed’ punctuation layout style. The first paragraph in any section of text is
(frequent commas at the end of clauses) or ‘open’ punc- usually not indented, but following paragraphs may be
tuation (less frequent punctuation) and be consistent. (by the equivalent of three character spaces). In addi-
tion, the space between paragraphs should be slightly
●● Do not link two sentences with a comma. Use a full
larger than the space between lines. Follow depart-
stop, this is an example of what not to do.
mental guidelines if these specify a format.
●● Pay special attention to apostrophes, using the follow-
●● Group paragraphs in sections under appropriate head-
ing rules:
ings and subheadings to reinforce the structure under-
– To indicate possession, use an apostrophe before an lying your writing.
‘s’ for a singular word (e.g. the solution’s temperature
was . . . ’) and after the s for a plural word ending in s ●● Think carefully about the first and last paragraphs in
(e.g. the solutions’ temperatures were = the temper- any piece of writing: these are often the most impor-
atures of the solutions were). If the word has a special tant as they respectively set the aims and report the
plural (e.g. woman S women) then use the apostro- conclusions.
phe before the s (the women’s temperatures were . . . ).
Note: If you are not sure what is meant by any of the
–
When contracting words, use an apostrophe terms used here, consult a guide on writing (see sources
(e.g.do not = don’t; it’s = it is), but remember that for further study).

Box 68.2 Using appropriate writing styles for different purposes (with examples)
Note that courses tend to move from assignments that To use this style, first decide on those aspects you wish
are predominantly descriptive in the early years to a to compare and then consider the material (e.g. different
more analytical approach towards the final year (see literature sources) from these aspects – in what ways do
Chapter 63). Also, different styles may be required in dif- they agree or disagree with each other? One approach is to
ferent sections of a write-up, e.g. descriptive for intro- compare/contrast a different aspect in each paragraph. At a
ductory historical aspects, becoming more analytical in practical level, you can use ’linking’ words and phrases to
later sections. help orientate your reader, as you move between aspects
where there is agreement and disagreement. These
Descriptive writing include, for agreement: ‘in both cases’; ‘in agreement
This is the most straightforward style, providing fac- with’; ‘is also shown by the study of’; ‘similarly’; ‘in the
tual information on a particular subject and is most same way’, and for disagreement: ‘however’; ‘although’;
appropriate: ‘in contrast to’; ‘on the other hand’; ‘which differs from’.The
comparative style is fairly straightforward, once you have
●● in essays where you are asked to ‘describe’ or ‘explain’ decided on the aspects to be compared.The following brief
(p. 599) example compares two different studies using this style:
●● when describing the results of a practical exercise, ‘While Black and White (2009) reported high levels of Pb
e.g.: ‘The experiment shown in Figure 1 confirmed that in 75% of the soil samples, previous work by Grey and
the reaction was strongly influenced by temperature, Gray (2005) indicated Pb levels 72000 mg kg-1 in selected
as the rate observed at 80°C was more than double “hot-spots” on the same site’.
that seen at 20°C ’.
Comparative text typically makes use of two or more ref-
However, in literature reviews and essays where erences per paragraph.
you are asked to ‘discuss’ (p. 599) a particular topic,
the descriptive approach is mostly inappropriate, as Analytical writing
in the following example, where a large amount of Typically, this is the most appropriate form of writing for:
specific information from a single scientific paper has
●● a review of scientific literature on a particular topic;
been used, without any attempt to highlight the most
important points: ●● an essay where you are asked to ‘discuss’ (p. 599) dif-
ferent aspects of a particular topic;
‘In a study carried out between July and October 2008,
a total of 250 soil samples were collected, stored and ●● evaluating a number of different published sources
then subsequently analysed. The results obtained indi- within the Discussion of a final-year project dissertation.
cated that high levels ( 7150 mg/kg) of Pb were obtained By considering the significance of the information pro-
in 75% of the samples; negligible levels of Cd, i.e. vided in the various sources you have read, you will be
60.1 mg/kg, were found in the same samples (Black and able to take a more critical approach. Your writing should
White, 2009)’. evaluate the importance of the material in the context of
your topic (see also Chapter 70). In analytical writing, you
In the most extreme examples, whole paragraphs or need to demonstrate critical thinking (p. 386) and per-
pages of essays may be based on descriptive factual sonal input about the topic in a well-structured text that
detail from a single source, often with a single citation at provides clear messages, presented in a logical order and
the end of the material, as above: such essays often score demonstrating synthesis from a number of sources by
low marks in essays where evidence of deeper thinking is appropriate use of citations (p. 379). Detailed informa-
required (Chapter 63). tion and relevant examples are used only to explain or
develop a particular aspect, and not simply as ’padding’
Comparative writing
to bulk up the essay. The following example shows how
This technique is an important component of academic detail can be used for explanation:
writing, and it will be important to develop your compar-
‘Historically, lead has been mined in selected areas in
ative writing skills as you progress through your course.
Northern England: a limited study on this site identified
Its applications include:
elevated lead levels in 188 samples (Black and White,
●● answering essay questions and assignments of the 2009), while a previous study had identified significant
‘compare and contrast’ type (p. 599) lead levels in isolated samples (Grey and Gray, 2005)’.
●● comparing your results with previously published Analytical writing is based on a broad range of sources,
work in the Discussion section of a practical report. typically with several citations per paragraph.

Box 68.3 How to improve your writing ability by consulting a personal reference library
Using dictionaries ●● Use a thesaurus to find a more precise and appropri-

ate word to fit your meaning, but check definitions of
We all know that a dictionary helps with spelling and
unfamiliar words with a dictionary.
definitions, but how many of us use one effectively?
You should: ●● Use it to find a word or phrase ‘on the tip of your
tongue’ by looking up a word of similar meaning.
●● Keep a dictionary beside you when writing and always
use it if in any doubt about spelling or definitions. ●● Use it to increase your vocabulary.
●● Use it to prepare a list of words which you have diffi- Roget’s Thesaurus is the standard. Several publishers
culty in spelling: apart from speeding up the checking produce a combined dictionary and thesaurus.
process, the act of writing out the words helps commit
them to memory. Using guides for written English
●● Use it to write out a personal glossary of terms. This These provide help with the use of words.
can help you memorise definitions. From time to time, ●● Use guides to solve grammatical problems such as
test yourself. when to use ‘shall’ or ‘will’, ‘which’ or ‘that’, ‘effect’ or
Not all dictionaries are the same! Ask your tutor or ‘affect’, ‘can’ or ‘may’, etc.
supervisor whether he/she has a preference and why. Try ●● Use them for help with the paragraph concept and the
out the Oxford Advanced Learner’s Dictionary, which is correct use of punctuation.
particularly useful because it gives examples of use of all
words and helps with grammar, e.g. by indicating which ●● Use them to learn how to structure writing for different
prepositions to use with verbs. tasks.
Using a thesaurus
A thesaurus contains lists of words of similar meaning
grouped thematically; words of opposite meaning always
appear nearby.
Reviewing and revising your text

Detailed review of each draft is strongly advised for all writing, apart from
in exams. When using a word processor, this can be a simple process. Where
possible, schedule your writing so you can leave each draft to ‘settle’ for at least
a couple of days. When you return to it fresh, you will see more easily where
improvements can be made. Try the following structured review process, each
stage being covered in a separate scan of your text:
Reviewing your text – to improve clar- 1. Examine content. Have you included everything you need to? Is all the
ity and shorten your text, ‘distil’ each material relevant?
sentence by taking away unnecessary 2. Check grammar and spelling. Can you spot any ‘howlers’?
words and ‘condense’ words or phrases 3. Focus on clarity. Is the text clear and unambiguous? Does each sentence
by choosing a shorter alternative. really say what you want it to say?
4. Be succinct. What could be missed out without spoiling the essence of
your work? It might help to imagine an editor has set you the target of
Learning from others – ask a colleague reducing the text by 15%.
to read through your draft and comment 5. Improve style. Could the text read better? Consider the sentence and par-
on its content and overall structure. agraph structure and the way your text develops to its conclusion.
Common errors
These include:
●● Problems over singular and plural words (‘the data is’; ‘the results show’)

●● Verbose text (‘One definition that can be employed in this situation is given
in the following sentence’.)
●● Misconstructed sentences (‘Health and safety regulations should be made
aware of . . . ’)
●● Misuse of punctuation, especially commas and apostrophes (for examples,
see Box 68.1).
●● Poorly constructed paragraphs (for advice, see Box 68.1).

Butterfield, J. (ed.) (2004) Fowler’s Modern English Usage, [This is excellent for the basics of English – it covers
revised 4th edn. Oxford University Press, Oxford. grammar, usage and the construction of sentences and
Clark, R. The English Style Book. A Guide to the Writing of paragraphs].
Scholarly English. Available: http://www.litencyc.com/ Lindsay, D. (2011) Scientific Writing = Thinking in Words,
stylebook/stylebook.php CSIRO, Collingwood.
Last accessed 01/09/15. McMillan, K.M. and Weyers, J.D.B. (2009) The Smarter
Day, R.A. and Sakaduski, N (2011) Scientific English: A Study Skills Companion. Pearson, Harlow.
Guide for Scientists and Other Professionals. Green- McCormac, C., Davis, J., Papakonstantinou, P. and Ward,
wood, Westport. N. (2012) Research Project Success. The essential guide
Holst, B. (2015) Scientific paper writing: A survival guide. for science and engineering students. Royal Society of
Createspace & Independent, London. Chemistry, Cambridge.
Kane, T.S. (2000) The Oxford Essential Guide to Writing.
Oxford University Press, New York.
Study exercises
68.1 ‘Brainstorm’ an essay title. Pair up with a part- ●● structure and flow;
ner in your class. Together, pick a suitable essay ●● vocabulary.
title from a past exam paper. Using the spider
Now either borrow a book from a library or
diagram or another technique, individually
buy a book that deals with your weakest aspects
‘brainstorm’ the title. Meet afterwards, compare
of writing. Read the relevant chapters or sections
your ideas, and discuss their relative merits and
and for each aspect write down some tips that
disadvantages.
should help you in future.
68.2 Improve your writing technique. From the fol-
68.3 Improve your spelling and vocabulary with two
lowing checklist, identify the three weakest
lists. Create a pair of lists and pin these up beside
aspects of your writing, either in your own opin-
your desk. One should be entitled Spelling Mis-
ion or from essay/assignment feedback:
takes and the other New Words. Now, whenever
●● grammar; you make a mistake in spelling or have to look up
●● paragraph organisation; how to spell a word in a dictionary, add the prob-
●● presentation of work; lem word to your spelling list, showing where
●● punctuation; you made the mistake. Also, whenever you come
●● scientific style; across a word whose meaning is unclear to you,
●● sentence structure/variety; look it up in a dictionary and write the word and
●● spelling; its meaning in the ‘new words’ vocabulary list.

69 Writing essays
1. read question The main functions of an essay are to show how much you understand about a
5. review answer and understand topic and how well you can organise and express your knowledge.
(10%) (5%)
2. research Organising your time

possible content
(20%) The way you should divide your time when producing an essay depends on
whether you are writing it for in-course assessment or under exam conditions
(Fig. 69.1). Essays written over a long period with access to books and other
resources will probably involve a research element, not only before the planning
phase but also when writing (Fig. 69.1(a)). For exams, it is assumed that you have
revised appropriately (Chapter 63) and essentially have all the information at your
fingertips. To keep things uncomplicated, the time allocated for each essay should
be divided into three components – planning, writing and reviewing (Fig. 69.1(b)),
and you should adopt time-saving techniques whenever possible (Box 64.2).
3. plan answer
(5%)
Making a plan for your essay
4. write essay, Dissect the meaning of the essay question or title
using sources
(a) (60%)
Read the title very carefully and think about the topic before starting to write.
Consider the definitions of each of the important nouns (this can help in
3. read answer and 1. read question and
make corrections plan answer
approaching the introductory section). Also think about the meaning of the
(5%) (10%) verb(s) used and try to follow each instruction precisely (see Table 69.1). Do
not get side-tracked because you know something about one word or phrase
in the title: consider the whole title and all its ramifications. If there are two or
more parts to the question, make sure you give adequate attention to each part.
Consider possible content and examples
Research content using the methods described in Chapters 41 and 42. If you
have time to read several sources, consider their content in relation to the essay
title. Can you spot different approaches to the same subject? Which do you
prefer as a means of treating the topic in relation to your title? Which examples
are most relevant to your case, and why?
Construct an outline
Every essay should have a structure related to its title.
(b) 2. write answer (85%) KEY POINT Most marks for essays are lost because the written
material is badly organised or is irrelevant. An essay plan, by
Fig. 69.1 Typical division of time for an
essay written as part of an in-course assess-
definition, creates order and, if thought about carefully, should
ment (a) or under exam conditions (b). ensure relevance.
Considering essay content – it is rarely Your plan should be written down (but scored through later if written in an
enough simply to lay down facts for exam book). Think about an essay’s content in three parts:
the reader – you must analyse them
and comment on their significance (see
1. The introductory section, in which you should include definitions and
p. 591).
some background information and the context for the topic being consid-
ered. You should also tell your reader how you plan to approach the subject.
2. The middle of the essay, where you develop your answer and provide rel-
evant examples. Decide whether a broad analytical approach is appropriate
or whether the essay should contain more factual detail.

Writing essays
Table 69.1 Instructions often used in essay questions and their meanings.
Ten Golden Rules for essay writing – When more than one instruction is given (e.g. compare and contrast;
these are framed for in-course assess- describe and explain), make sure you carry out both or you may lose a
ments (p. 560), though many are also large proportion of the available marks (see also Table 64.1).
relevant to exams (see also Box 64.2).
Account for: give the reasons for
1. Read the question carefully, and Analyse: examine in depth and describe the main characteristics of
decide exactly what the assessor Assess: weigh up the elements of and arrive at a conclusion about
wants you to achieve in your answer. Comment: give an opinion on and provide evidence for your views
2. Make sure you understand the Compare: bring out the similarities between
question by considering all Contrast: bring out dissimilarities between
aspects – discuss your approach Criticise: judge the worth of (give both positive and negative aspects)
with colleagues or a tutor. Define: explain the exact meaning of
Describe: use words and diagrams to illustrate
3. Carry out the necessary research
Discuss: provide evidence or opinions about, arriving at a balanced
(using books, journals, the web.) conclusion
taking appropriate notes. Gain an Enumerate: list in outline form
overview of the topic before getting Evaluate: weigh up or appraise; find a numerical value for
involved with the details. Explain: make the meaning of something clear
4. Always plan your work in outline Illustrate: use diagrams or examples to make clear
before you start writing. Check that Interpret: express in simple terms, providing a judgement
your plan covers the main points Justify: show that an idea or statement is correct
List: provide an itemised series of statements about
and that it flows logically.
Outline: describe the essential parts only, stressing the classification
5. Introduce your essay by show-
Prove: establish the truth of
ing that you understand the topic Relate: show the connection between
and stating how you intend to Review: examine critically, perhaps concentrating on the stages in the
approach it. development of an idea or method
6. As you write the main content, State: express clearly
ensure it is relevant by continually Summarise: without illustrations, provide a brief account of
looking back at the question. Trace: describe a sequence of events from a defined point of origin
7. Use headings and sub – headings to
organise and structure your essay. 3. The conclusion, which you can make quite short. You should use this
8. Support your statements with rele- part to summarise and draw together the components of the essay, without
vant examples, diagrams and refer- merely repeating previous phrases. You might mention such things as: the
ences where appropriate. broader significance of the topic; its future; its relevance to other important
9. Conclude by summarising the key areas of biology. Always try to mention both sides of any debate you have
points of the topic, indicating the touched on, but beware of ‘sitting on the fence’.
present state of knowledge, what
we still need to find out and how
this might be achieved. KEY POINT Use paragraphs to make the essay’s structure obvi-
10. Always review your essay before ous. Emphasise them with headings and sub – headings unless
submitting it. Check grammar the material beneath the headings would be too short or trivial.
and spelling and confirm that you
have answered all aspects of the
Start writing
question.
●● Never lose track of the importance of content and its relevance. Repeat-
edly ask yourself: ‘Am I really answering this question?’ Never waffle just
to increase the length of an essay. Quality, rather than quantity, is important.
Using diagrams – give a title and legend ●● Illustrate your answer appropriately. Use examples to make your points
to each diagram so that it makes sense in clear, but remember that too many similar examples can stifle the flow of an
isolation and point out in the text when essay. Use diagrams where a written description would be difficult or take
the reader should consult it (e.g. ‘as too long. Use tables to condense information.
shown in Fig. 1 . . . ’ or ‘as can be seen in
the accompanying diagram, . . . ’).
●● For handwritten essays, take care with your handwriting. You can-
not get marks if your writing is illegible. Try to cultivate an open form of

Writing essays
handwriting, making the individual letters large and distinct. If there is time,
Learning from lecturers’ and tutors’
make out a rough draft from which a tidy version can be copied.
comments – ask for further explanations
if you do not understand a comment or
Reviewing your answer
why an essay was less successful than
you thought it should have been. Make sure that you leave enough time to:
●● re-read the question to check that you have answered all points;
●● re-read your essay to check for errors in punctuation, spelling and content.
Make any corrections obvious. In an exam, do not panic if you suddenly
realise you have missed a large chunk out as the reader can be redirected to
a supplementary paragraph if necessary.

Good, S. and Jensen, B. (1995) The Student’s Only Survival McMillan, K.M. and Weyers, J.D.B. (2011) How to write
Guide to Essay Writing. Orca Book Publishers, Victoria. Essays and Assignments, 2nd edn. Pearson Education,
McCormac, C., Davis, J., Papakonstantinou, P. and Ward, London.
N. (2012) Research Project Success. The essential guide Shields, M. (2010) Essays Writings: A Student’s Guide.
for science and engineering students. Royal Society of Sage Publications, London.
Chemistry, Cambridge.
Study exercise
69.1 Practise dissecting essay titles. Use past exam (a) Print out a copy of the essay. Do not look at it
papers, or make up questions based on the learn- for at least two days after finishing this version.
ing outcomes for your course and your lecture (b) Review 1: spelling, grammar and sense. Read
notes. Take each essay title and carefully ‘dissect’ through the draft critically (try to imagine it
the wording, working out exactly what you think had been written by someone else) and cor-
the assessor expects you to do (see e.g.Table 69.1). rect any obvious errors that strike you. Does
69.2 Write essay plans under self-imposed time lim- the text make sense? Do sentences/ para-
its. Continuing from study exercise 69.1, outline graphs flow smoothly?
plans for essays from a past exam paper. Allow (c) Review 2: structure and relevance. Consider
yourself a maximum of 5 min per outline. Within again the structure of the essay, asking your-
this time your main goal is to create an essay self whether you have really answered the
plan. To do this, you may need to ‘brainstorm’ question that was set (see study exercise
the topic. Alternatively, if you allocate 10 min per 69.1). Are all the parts in the right order? Is
essay, you may be able to provide more details, anything missed out? Have you followed pre-
e.g. list the examples you could describe. cisely the instruction(s) in the title? Are the dif-
ferent parts of the essay linked together well?
69.3 Practise reviewing your work carefully. For the
(d) Review 3: shorten and improve style. Check
next assignment you write, review it fully as part of
the word count. Shorten the essay if required.
the writing process. This will require you to finish
Look critically at phrasing and, even if the
the first draft about one week before the hand-in
essay is within the word limit, ask yourself
date, e.g. by setting yourself an earlier deadline
whether any of the words are unnecessary
than the submission date. This exercise is best
or whether the text could be made more con-
done with a word-processed essay. Do not worry
cise, more precise or more apt.
if it is a little over the word limit at this stage.
Answers to these study exercise are available at www.pearsoned.co.uk/practicalskills.

70 Reporting practical and project work
Practical reports, project reports, theses and scientific papers differ greatly in
Typicalx structure of scientific
depth, scope and size, but they all have the same basic structure. Some variation
reports – this usually follows the
is permitted, however (see Box 70.1), and you should always follow the advice
‘IMRaD’ acronym: Introduction, Mate-
or rules provided by your department.
rials and Methods, Results and Dis-
Additional parts may be specified: for project reports, dissertations and
cussion. OR the ‘IERaD’ acronym:
theses, a Title page is often required and a List of Figures and Tables as part
Introduction, Experimental, Results and
of the Contents section. When work is submitted for certain degrees, you may
Discussion.
need to include certain declarations and statements made by the student and
supervisor. In scientific papers, a list of Key Words is often added following
the Abstract: this information may be combined with words in the title for
computer cross-referencing systems.
Key Point Department, school or faculty regulations may

specify a precise format for your report or thesis. Consult a copy
of these rules at an early stage and follow them closely, to avoid
losing marks.
Practical laboratory reports

Options for discussing data – the main
optional variants of the general struc- These are exercises designed to make you think more deeply about your experi-
ture include combining Results and Dis- ments and to practise and test the skills necessary for writing up research work.
cussion into a single section and adding Special features are:
a separate Conclusions section. ●● Introductory material is generally short and, unless otherwise specified,
●● The main advantage of a joint should outline the aims of the experiment(s) with a minimum of background
Results and Discussion section is material.
that you can link together different
experiments, perhaps explaining
●● Materials and methods may be provided by your supervisor for practi-
why a particular result led to a new cal reports. If you make changes to this, you should state clearly what you
hypothesis and the next experiment. did. With extended projects your lab notebook (see p. 53) should provided
However, a combined Results and the basis for writing this section.
Discussion section may contravene
●● Great attention in assessment will be paid to presentation and analysis
your department’s regulations, so
you should check before using this
of data. Take special care over graphs (see p. 451 for further advice). Make
approach. sure your conclusions are justified by the evidence you present.
●● The main advantage of having a sep-
arate Conclusions section is to draw Theses and dissertations
together and emphasise the chief These are submitted as part of the examination for a degree following an
points arising from your work, when extended period of research. They act to place on record full details about
these may have been ‘buried’ in an
your experimental work and will normally only be read by those with a direct
extensive Discussion section.
interest in it – your examiners or colleagues. Note the following:
●● The introduction and Experimental sections are lengthy detailed, pro-
viding the reader with the full context for your research.
●● You are allowed scope to expand on your findings and to include detail
Oral assessments – there may be an that might otherwise be omitted in a scientific paper.
oral exam (viva voce) associated with
the submission of a thesis or disserta-
●● You may have problems with the volume of information that has to be
tion. The primary aim of the examiners
organised. One method of coping with this is to divide your thesis into
will be to ensure that you understand
chapters, each having the standard format (as in Box 70.1). A General Intro-
what you did and why you did it.
duction can be given at the start and a General Discussion at the end. Discuss
this with your supervisor.

Reporting practical and project work
Box 70.1 The structure of reports of experimental work using the ‘IERaD’ structure
Undergraduate practical and project reports are generally modelled on this arrangement or a close variant of it,
because this is the structure used for nearly all research papers and theses. The more common variations include
Results and Discussion combined into a single section and Conclusions appearing separately as a series of points
arising from the work. In scientific papers, a list of Key Words (for computer cross-referencing systems) may be
included following the Abstract. Acknowledgements may appear after the Contents section, rather than near the end.
Department or faculty regulations for producing theses and reports may specify a precise format; they often require a
Title page to be inserted at the start and a List of Figures and Tables as part of the Contents section, and may specify
declarations and statements to be made by the student and supervisor.
Part (in order) Contents/purpose Checklist for reviewing content
Title Explains what the project was about Does it explain what the text is about succinctly?
Authors plus their Explains who did the work and where; Are all the details correct?
institutions also where they can be contacted now
Abstract/Summary Synopsis of methods, results and conclu- Does it explain why the work was done?
sion of work described. Allows the reader Does it outline the whole of your work and your findings?
to grasp quickly the essence of the work
List of Contents Shows the organisation of the text (not Are all the sections covered?
required for short papers) Are the page numbers correct?
Abbreviations Lists all the abbreviations used (but not Have they all been explained?
those of SI, chemical elements or stand- Are they all in the accepted form?
ard chemical terms) Are they in alphabetical order?
Introduction Orientates the reader, explains why the Does it provide enough background information and cite all the
work has been done and its context in relevant references?
the literature, why the methods used Is it of the correct depth for the readership?
were chosen, why the experimental con- Have all the technical terms been defined?
ditions were chosen. Indicates the central Have you explained why you investigated the problem?
hypothesis behind the experiments Have you outlined your aims and objectives?
Have you explained your methodological approach?
Have you stated your hypothesis?
Experimental Explains how the work was done. Should Is each experiment covered and have you avoided unnecessary
contain sufficient detail to allow another duplication?
competent worker to repeat the work Is there sufficient detail to allow repetition of the work?
Have you explained where you got them from?
Are the correct names, sources and grades given for all chemicals?
Results Displays and describes the data Is the sequence of experiments logical? Are the parts adequately
obtained. Should be presented in a form linked?
which is easily assimilated (graphs rather Are the data presented in the clearest possible way?
than tables, small tables rather than Have SI units been used properly throughout?
large ones) Has adequate statistical analysis been carried out?
Is all the material relevant?
Are the figures and tables all numbered in the order of their
appearance? Are their titles appropriate?
Do the figure and table legends provide all the information neces-
sary to interpret the data without reference to the text?
Have you presented the same data more than once?
Discussion/ Discusses the results: their meaning, Have you explained the significance of the results?
Conclusions their importance; compares the results Have you compared your data with other published work?
with those of others; suggests what to Are your conclusions justified by the data presented?
do next
Acknowledgements Gives credit to those who helped carry Have you listed everyone that helped, including any grant-
out the work awarding bodies?
Literature Cited Lists all references cited in appropriate Do all the references in the text appear on the list?
(Bibliography) format: provides enough information to Do all the listed references appear in the text?
allow the reader to find the reference in Do the years of publications and authors match?
a library Are the journal details complete and in the correct format?
Is the list in alphabetical order, or correct numerical order?

Steps in the production of a practical report research

project or thesis
Choose the observations or the experiments you wish to describe
Choosing between graphs and and decide how best to present them
tables – graphs are generally easier Try to start this process before your lab work ends, because at the stage of
for the reader to assimilate, while tables reviewing your experiments, a gap may become apparent (e.g. a missing control)
can be used to condense a lot of data and you might still have time to rectify the deficiency. Irrelevant material should
into a small space. be ruthlessly eliminated, at the same time bearing in mind that negative results
can be extremely important (see p. 46). Use as many different forms of data pres-
entation as are appropriate, but avoid presenting the same data in more than one
form. Relegate large tables of primary data to an appendix and summarise the
Repeating your experiments – remem-
ber, if you do an experiment twice, you
important points within the main text (with a cross-reference to the appendix).
have repeated it only once.
Make sure that the experiments you describe are representative: always state
the number of times they were repeated and how consistent your findings were.
Make up plans or outlines for the component parts
The overall structure of practical and project reports is well defined (see
Presenting your results – remember Box 70.1), but individual parts will need to be well organised, as with any
that the order of results presented in other form of writing (see Chapter 68).
a report need not correspond with the Write
order in which you carried out the exper-
The Experimental (Materials and Methods) section is often the easiest to write
iments: you are expected to rearrange
once you have decided what to report. Remember to use the past tense and do
them to provide a logical sequence of
not allow results or discussion to creep in. There may be minor variations in
findings.
style but the are shown in Box 70.2 will be suitable in the majority of cases. The
Results section is the next easiest as it should only involve description. At this
stage, you may benefit from jotting down ideas for the Discussion – this may
Using the correct tense – always use be the hardest part to compose as you need an overview both of your own work
the past tense to describe the method- and of the relevant literature. It is also liable to become wordy, so try hard to
ology used in your work, since it is now make it succinct. The Introduction should not be too difficult if you have fully
complete. Use the present tense only understood the aims of the experiments. Write the Abstract and complete the list
for generalisations and conclusions. of references at the end. To assist with the latter, it is a good idea as you write to
collate appropriately formatted details of references (Chapter 9) as you progress.
Revise the text
Preparation is key – gather all relevart
Once your first draft is complete, try to answer all the questions given in
research papers in advance and collate
Box 70.1. Show your work to your supervisors and learn from their comments.
your data (tables, figures and schemes).
Let a friend or colleague who is unfamiliar with your subject read your text;
they may be able to pinpoint obscure wording and show where information or
explanation is missing. If writing a thesis, doublecheck that you are adhering
Research report or thesis – ensure to your institution’s thesis regulations.
your completed research report or the- Prepare the final version
sis fulls within your university’s word
Markers appreciate neatly produced work but a well-presented document will not
limit.
disguise poor science. If using a word processor, print the final version with the
best printer available. Make sure figures are clear and in the correct size and format.
Submit your work
Thesis – you have been researching
Your department will specify when to submit a thesis or project report, so plan your
your chosen subject for several years
work carefully to meet this deadline or you may lose marks. Tell your supervisor
and you know lots of information about
early of any circumstances that may cause delay. Also, check to see whether any
it. You just have to write it down.
forms are required for late submission, or evidence of extenuating circumstances.
Guidance or how to write up a research project or thesis is shown in Box 70.3.

Box 70.2 Writing experimental procedures
The main purpose of the methods described in the exper- Experimental

imental section of a laboratory report, project report, the- A mixture of 2,4-dimethylbenzoic acid (3.0 g, 0.02 mol),
sis or paper is to communicate sufficient information to dimethylsulphate (2.8 g, 0.022 mol) and anhydrous potas-
allow an experienced chemist to repeat your experiments. sium carbonate (3.3 g, 0.024 mol) in dry propanone (20mL)
One of the goals of writing laboratory reports is to pro- was boiled under reflux (3h), cooled, poured into water
vide you with practice in writing in the generally accepted (100 mL) and extracted with dichloromethane (3 * 20 mL).
style required for more professional publications. The fol- Removal of the dried (MgSO4) organic solvent gave a
lowing points should be noted: white solid (2.3 g), which was recrystallised from ethanol,
●● Always write in the third person, past tense. using charcoal to improve the colour, to yield white nee-
dles of methyl 2,4-dimethylbenzoate (2.28 g). The infrared
●● Do not copy word for word the instructions given in (Nujol) and 1H9NMR (CDCl3) spectra were recorded and
the experiment protocol. You are in a learning situa- TLC (SiO2/CH2Cl2) showed the product to be pure. The
tion in the laboratory and the protocol may contain melting point was determined and the % yield calculated.
information, which may be new to you, but is general
knowledge to experienced chemists. Note 1. Experimental gives reagents, solvents, quantities,
times, yields, etc., and sufficient detail for the experiment
●● Condensing the experimental protocol for your to be repeated by a proficient chemist. Details of standard
report will help you to see the important steps in the techniques such as weighing liquids, setting up apparatus,
experiment. recrystallisation, drying the product, etc., are ‘understood’
●● All sentences begin with a capital letter not a number by the experimentalist from experience and training.
or bracket.
Note 2. The weights and volumes used are quoted to one
The following examples illustrate the differences between decimal place, since this is the level of accuracy required
the instructions of the experiment protocol and the for preparative experiments which results from the tech-
accepted style required for the experimental section. niques used, distillation, recrystallisation, extraction, etc.,
and the non-quantitative conversion of the reactants to
Example 1 A preparative experiment: the synthesis of products may be more significant than errors in weighing.
methyl 2,4-dimethylbenzoate
Note 3. All analytical data such as IR and NMR spectra,
Experiment protocol TLC, melting point and yield calculations are entered in
the results section.
‘In a round-bottom flask (100 mL) place 2,4-dimethyl-
benzoic acid (3.0 g, 0.02 mol), anhydrous potassium car- Note 4. The % yield for the product represents a compari-
bonate (3.3 g, 0.024 mol) and a small magnetic flea. In the son with the theoretical yield of the reaction and the prac-
fume cupboard and wearing protective gloves, carefully tical yield. Calculation of the theoretical yield based on:
weigh out dimethylsulphate (2.8 g. 0.022 mol) into a glass
(a) the molar quantities of the reactants used;
sample tube (10 mL) using a Pasteur pipette to transfer the
liquid. Using the Pasteur pipette, transfer the dimethylsul- (b) the number of moles of each reactant required to
phate to the reaction flask and use anhydrous propanone make 1 mole of product;
(10 mL) to rinse the sample tube. Add the rinsings to the
reaction flask and add more dry propanone (10 mL). Fit the (c) the assumption that reactions go 100%;
flask with a reflux condenser and boil the mixture under (d) the ‘limiting quantity’ of one of the reactants.
reflux for 3 hours using an oil bath on a stirrer hot plate in
the fume cupboard. Allow the reaction mixture to cool to To calculate the theoretical yield of a reaction, it is essen-
room temperature and pour into water (100 mL) washing tial that you recognise the reactant, which is the limiting
the reaction flask with a few millilitres of water. Extract quantity.
the aqueous solution three times with dichloromethane If a reaction requires 1 mole of reactant A and 1 mole of
(20 mL), dry the dichloromethane (MgSO4), filter off the reactant B to give 1 mole of product AB, i.e.:
drying agent, remove the solvent on the rotary evaporator A + B S AB
and record the weight of the crude product. Recrystallise
and charcoal the crude ester from ethanol and dry in a if we use A (1 mol) and B (1.5 mol), then only 1 mol of
vacuum desiccator. Record the weight and melting point AB can be formed and there is 0.5 mol of B in excess and
of the purified ester and obtain and interpret infrared and unreacted. Therefore the limiting quantity is the amount
1
H9NMR spectra. Check the purity of your product using of reactant A (1 mol). The excess of reactant B may be
TLC (SiO2 plates and CH2Cl2 as eluent) and calculate the necessary to ensure complete conversion of A into AB,
percentage yield of the product’. i.e. 100% reaction. In the example above, the limiting

quantity is the amount of 2,4-dimethylbenzoic acid (0.02 B. Sodium thiosulphate (0.01 M)

mol) and only 0.02 mol of the methyl ester can be formed C. Potassium iodide (0.4 M).
(164 * 0.02 = 3.28 g). Therefore: i. Place solution A (100 mL) and solution C (100 mL)
reaction yield 2.28 into separate conical flasks and suspend them in
% yield = * 100 = * 100 = 70%
theory yield 3.28 a thermostat bath at 25 °C.
ii. Mix solutions A and C (50 mL of each) in a stop-
Example 2 A quantitative experiment: standardisation of
pered flask and immerse in hot water until the
sodium thiosulphate solution
end of the experiment.
Experiment protocol iii. After about 15 minutes from step i, mix the solu-
‘Prepare a standard solution of potassium iodate by dis- tions A and C and start the clock. This mixture
solving potassium iodate (about 1.34 g, accurately weighed) must be kept in the thermostat bath at 25 °C,
in distilled water (100 mL), quantitatively transferring the throughout the experiment.
solution to a volumetric flask (250.00 mL) making up to the iv. Just before 2 minutes have elapsed, take a sam-
mark using distilled water and mixing well. Pipette an ali- ple (10.00 mL) from the flask by pipette and add
quot (25.00 mL) of the solution into a conical flask (250 mL) it to distilled water (200 mL) in a separate flask.
and add sulphuric acid (50 mL, 1 M). Weigh out potassium This quenches (stops) the reaction.
iodide (1 g), add it to the conical flask and swirl until it has all v. Take further samples at 4, 6, 9, 13, 18, 24, 32, 44
dissolved. Titrate the liberated iodine (brown solution) with and 60 minutes and quench in the same way.
the sodium thiosulphate solution until a pale straw colour
vi. Titrate each quenched solution with sodium thio-
is reached. Add iodine indicator (two drops) or freshly pre-
sulphate, including a sample of the ‘hot-water’
pared starch solution (two drops) and continue the titration
sample. Use a small amount of freshly prepared
until the colour changes from blue–black to colourless. If the
starch indicator solution and titrate from blue to
solution does not turn blue–black when you add the iodine
colourless. The titres for the samples are the val-
indicator or starch, you have overshot the endpoint. Repeat
ues (T) and that for the ‘hot-water’ sample is T ∞ .
the experiment until consistent results are obtained. Calcu-
late the molarity of the sodium thiosulphate solution’. vii. Now repeat the whole experiment using a ther-
mostat bath set at 35–40 °C. Make an accurate
Experimental record of the temperature. You do not need to
Potassium iodate (1.3402 g) was dissolved in distilled water repeat the ‘hot-water’ sample since this will serve
(100 mL), transferred to a volumetric flask (250.00 mL) for both experiments. Since this higher temper-
and made up to the mark using distilled water. An aliquot ature reaction is faster, there is no need to take
(25.00 mL) was transferred to a conical flask (250 mL), samples at 44 and 60 minutes.
sulphuric acid (50 mL, 1 M) and potassium iodide (1 g) viii. For each temperature plot a graph of - ln(T ∞ - T)
were added and the mixture swirled to effect solution. The versus time (s)’.
iodine liberated was titrated with the sodium thiosulphate
solution to a pale straw colour. Iodine indicator (two drops) Experimental
was then added and the titration continued to a blue-black Solutions (100 mL) of potassium persulphate (0.04 M) and
to colourless endpoint. The experiment was repeated until potassium iodide (0.4 M) were equilibrated (0.25 h) in a
consistent results were obtained and the concentration of constant temperature bath (25 °C). The two solutions were
the sodium thiosulphate solution calculated. mixed in a conical flask (250 mL) in the constant tempera-
ture bath and the clock started. After 2 minutes, an aliquot
Note 1. The differences in accuracy, indicating the equip- (10.00 mL) of the mixture was quenched with distilled water
ment you must use, are shown by the decimal point (200 mL) and the sampling process repeated after 4, 6, 9,
quoted in the quantities. 13, 18, 24, 32, 44 and 60 minutes. Each sample was titrated
Note 2. Titrations should be repeated to give consecutive with sodium thiosulphate solution (0.01 M), using iodine
results to within one or two drops of titrant. Do not aver- indicator, to a colourless end-point and the values recorded
age widely differing results. (T).The experiment was then repeated at a known tempera-
ture between 35 °C and 40 °C but the samples at 44 and 60
Note 3. Balance readings, burette readings and calcula- minutes were not taken. Solutions (50 mL) of the potassium
tions should be included in the results section. persulphate and potassium iodide were mixed and kept in
Example 3 Reaction kinetics: determination of rate con- a water bath (60–70 °C) for 2 hours. An aliquot (10.00 mL) of
stant and energy of activation using titrimetry this solution was quenched in distilled water (200 mL) and
titrated (T ∞ ) with the sodium thiosulphate solution. A graph
Experiment protocol of -ln(t ∞ - T) versus time (s) was plotted.
‘The following solutions are provided: Note 1. Only experimental detail is given. Calculations,
A. Potassium persulphate (0.04 M) results and theory are in the appropriate sections.

Box 70.3 How to write up your research project (disseration) or thesis
1. Getting started. Writing a research project or the- at research projects or theses from previous students,
sis will probably constitute the biggest writing task ideally in your specific research area. In the case of
that you will have undertaken to date. It will be the theses, it is now very common that all past-successful
result of you undertaking a major research project in theses are available freely online in open access sites.
chemical sciences (e.g. synthetic chemistry, analyti- Prior to the commencement of the writing it is a good
cal chemistry, physical chemistry). While the layout idea to arrange a meeting with your research super-
of the research project or thesis can vary depending visor to discuss the content of your research project
upon your university, department or supervisor, they or thesis. Following this meeting map out an outline
generally follow a certain pattern (or style): Introduc- table of contents, including major chapter Headings,
tion, Experimental and Results & Discussion (Note: then sub-headings per chapter and any other detail
In some areas of chemistry (e.g. synthetic chemistry) you can add. At this draft stage it is a good idea to
the Experimental section can appear at the end of the discuss with your research supervisor again. It is also
research project or thesis.) The Results & Discussion important to realise that this activity so far is classed
may also be split into multiple chapters to allow a as ‘draft’. Ultimately when you start writing and add-
range of related topics to be discussed and their data ing content to each section you will start to appreciate
presented. In addition to these three main aspects whether the scientific ‘story’ you are seeking to tell is
additional information is also required, specifically in a logical and progressive order.
an abstract (one page), acknowledgements, bibliog- Practically a good place to start is in the Experi-
raphy (references) and appendices (contains details mental and Results & Discussion sections. It is not
not included in the Results and Discussion section). that writing these sections are easy but they do reflect
what you have actually done – so should come the
2. Selecting the environment. Prior to the start of the easiest. Often people will leave the Introduction sec-
writing process it is important to consider where you tion to later. The Introduction is about ‘scene setting’
are going to do it; the selected area needs to be com- for your research project or thesis and so by definition
fortable, and away from external distractions. It is it is a summary of what other researchers have done
also important to ensure you have all the necessary in your field of study. It will also contain a large num-
information to write the research project or thesis at ber of references that need to be cited correctly both
your disposal. Your chosen environment may well in the text and bibliography (Chapters 42–44).
be the university library, your university accommo- In all writing (research project or thesis) it is impor-
dation or at home. It is also important to consider tant to get some text down on the computer. Once
when you will work on your research project or thesis. you have something it is possible to be self-critical
This is down to personal preference but never the and subsequently adapt and modify the text. Often
less you need to select the timeframe to do it. For a good starting point for deciding what to write can
example, your preference may be late morning to be established by deciding what the content of fig-
early evening. What is important is that you quickly ures and tables (and in the case of synthetic chemis-
establish a regular routine and stick with it. Equally try reaction schemes) should be. Once this content is
important is that outside this dedicated ‘writing time’ established it is then a matter of writing about them.
you build in relaxation time; this might be meeting up
with friends, a walk or some sporting activity. 5. Writing up. It is extremely important to ensure that all
your writings are appropriately electronically stored
3. Effective writing. You will be aware already because and backed up. There is nothing so disappointing and
of your past academic career that your concentration frustrating as to discover that the work you have writ-
to work effectively is time limited. In this context you ten up has been lost due to a computer malfunction or
need to maximise your creative writing time, i.e. writ- careless error on your part. So the backing up of your
ing the research project or thesis, alongside activities writings on your hard drive, a memory stick/external
that can support this writing. Such ‘support activities’ hard drive and the university server is essential. At this
might include the preparation of figures, tables and stage it is also very important to ensure you have some
schemes. It is also important during the writing stage coding to be able to identify what version of your files
to keep yourself hydrated and nourished: it is no use you are working on. This could be done by date order
feeling hungry or thirsty whilst writing – this is in or by adding the phrase ‘v1’ – for version 1 and so on.
itself a distraction.
6. Proofreading. Once you have drafted a chapter of
4. Where to start. It is important to identify a style that you your research project or thesis it is a good idea to
propose to follow to write up your research project or undertake some basic proofreading (by yourself). If
thesis prior to starting.This can be achieved by looking possible put your ‘objective glasses’ on and see if

you could improve the text, tables and figures (and academic members of staff, one of whom is likely to
schemes) in any way. Be vigilant for some obvious be your research supervisor alongside an independ-
errors, e.g. check the text is in the same font and ent assessor. They will each independently assess
size (your university or department may well have your research project and then agree an appropriate
guidelines on this so ensure you follow them). Once mark (or grade). This mark (or grade) will be collated
you have corrected your known errors it is advisable by the module tutor who will submit the mark for ver-
to ask someone else to check the writing for you. ification by an external examiner at the final exami-
This could be a fellow student who is knowledgeable nation board when your degree classification will be
about your subject as well as a non-specialist. Each confirmed (and awarded). A research project often
can contribute a different perspective; ultimately contributes a significant proportion of your overall
however it is your decision as to what to include or final degree classification (or grade). Typically, this
not. Once this process has taken place it is important may range between 25% for a BSc project and 50%
to get feedback from your research supervisor who for an MChem.
can comment on style, layout and content. For thesis only – Once your thesis has been submitted
After seeking advice it is important that you consider a period of up to 2 months may go by before the final
what changes you are actually going to make. All your viva voce examination (or ‘viva’) takes place. During
‘advisers’ have acted in your best interest – that is why this time two independent examiners (at least one of
you asked them in the first place. However, perhaps who is external to the university) read your thesis,
the most important is the research supervisor as they and the organisation of the viva is taking place. This
will probably be part of the assessment process (for time provides you with the opportunity to review your
an undergraduate research project). Their experience thesis, pre-empt possible questions that you might
in academic writing is invaluable to you in this process. be asked and have a mock viva with your supervisor
7. Bibliography. The bibliography lists all the refer- (or other designated staff). It is also wise to find out
ences you have used throughout the writing of your about your examiners – what is their specific area of
research project or thesis. The university, department research, what have they published in their research
or supervisor may well have a preferred style for how field recently.
references must be cited. What is sacrosanct is that all It is important in this post-submission stage that
references must be written in a single and consistent you relax but keep focused on the task to come. Prior
style, against an established system. Chapter 41 pro- to the day you will be contacted with details of the
vides details for the citing of published information. room for the viva, the date and time when you should
arrive. It is likely when you enter the viva you will find
8. The final draft. It is highly probable that you will pro-
both examiners and an independent non-examining
duce multiple drafts of your research project or thesis
chair (someone appointed by the university to over-
until you get to a position that it is done. Deciding it is
see the formal procedures of the viva). The examiners
done and complete can be a significant and difficult
will ask you to ‘defend’ your thesis by answering their
decision to make. While a deadline is critical it is also
questions. This whole process takes place over a few
important that you time manage (Chapter 59) your
hours (typically 2 hours).
writing to optimise the final research project or thesis.
Post-viva you will be invited to leave the room
Having a final proofread is crucial prior to submis-
while the examiners discuss the outcome. They
sion. Be aware of inconsistences in text, figures and
will then invite you back in to the room and ver-
tables (as well as reaction schemes). It is important,
bally confirm their recommendation (to award a
at this stage, that you also provide a copy to your
research degree). It is highly probable that they
research supervisor for final comments. Finally, after
will require some corrections to the thesis prior
making any last modifications ensure the finished
to final award. It is extremely important that these
research project or thesis is stored electronically with
corrections are made to the thesis in consultation
a suitable label (and in multiple locations).
with your research supervisor. It is also the last
9. Submission. Universities (and departments) will chance to eliminate any minor errors that you may
have different procedures for the submission of your be aware of.
research project or thesis. Ensure that you are aware Universities have specific procedures for the sub-
of deadlines and the process of submission well in mission of the final thesis which must be adhered to.
advance so that you can time manage this event (and This will include details on exact layout, the number
not feel unduly pressurised). of copies to be submitted, the format for binding of
the thesis as well details of submission of an elec-
10. After submission.
tronic ‘pdf’ copy. This latter pdf copy will usually be
For research projects only – Once your research pro- submitted by the university to an open access site for
ject is submitted it will be assessed, often by two public viewing.

Producing a scientific paper

Definition
Scientific papers are the means by which research findings are communi-
Peer review – the process of evalua- cated to others. Peer-reviewed papers are published in journals; each covers a
tion and review of a colleague’s work. well-defined subject area and publishes details of the format they expect.
In scientific communication, a paper is
reviewed by two or more expert review- Key Point Peer review is an important component of the pro-
ers (‘referees’) for comments on quality cess of scientific publication; only those papers whose worth is
and significance as a key component of confirmed by the peer review process will be published.
the validation procedure.
It would be very unusual for an undergraduate to submit a paper on his or her

own – this would normally be done in collaboration with your project supervi-
sor, and only then if your research has satisfied appropriate criteria. However,
it is important to understand the process whereby a paper comes into being
(Box 70.4), as this can help you understand and interpret the primary literature.
Box 70.4 Steps in producing a scientific paper
Scientific papers are the lifeblood of any science and it is Writing

a major landmark in your scientific career to publish your
The paper’s format will be similar to that shown in Box 70.1
first paper. The main steps in doing this should include
and the process of writing will include outlining, review-
the following:
ing, etc., as discussed elsewhere in this chapter. Figures
Assessing potential content must be finished to an appropriate standard and this may
involve preparing photographs or digital images of them.
The work must be of an appropriate standard to be pub-
lished and should be ‘new, true and meaningful’.Therefore, Submitting
before starting, the authors need to review their work criti-
When completed, copies of the paper are submitted to
cally under these headings.The material included in a scien-
the editor of the chosen journal with a simple cover-
tific paper will generally be a subset of the total work done
ing letter. A delay of one to two months usually follows
during a project, so it must be carefully selected for rele-
while the manuscript is sent to two or more anonymous
vance to a clear central hypothesis – if the authors will not
referees who will be asked by the editor to check that the
prune, the referees and editors of the journal certainly will.
paper is novel, scientifically correct and that its length
Choosing a journal is justified.
There are thousands of journals covering biology and Responding to referees’ comments
each covers a specific area (which may change through
The editor will send on the referees’ comments to the
time). The main factors in deciding on an appropriate
authors, who will then have a chance to respond. The
journal are the range of subjects it covers, the quality of
editor will decide on the basis of the comments and
its content and the number and geographical distribution
replies to them whether the paper should be published.
of its readers. The choice of journal always dictates the
Sometimes quite heated correspondence can result if the
format of a paper since authors must follow to the letter
authors and referees disagree.
the journal’s ‘Instructions to Authors’.
Checking proofs and waiting for publication
Deciding on authorship
If a paper is accepted, it will be sent for production.
In multi-author papers, a contentious issue is often who
The next the authors see of it is the proofs (first printed
should appear as an author and in what order they should
version in style of journal), which have to be checked
be cited. Where authors make an equal contribution, an
carefully for errors and returned. Eventually, the paper
alphabetical order of names may be used. Otherwise,
will appear in print, but a delay of six months following
each author should have made a substantial contribu-
acceptance is not unusual. Nowadays, papers are often
tion to the paper and should be prepared to defend it in
available electronically, via the web, in PDF format – see
public. Ideally, the order of appearance will reflect the
p. 384 for advice on how to cite such ‘online early’ papers
amount of work done rather than seniority. This may not
using the DOI system.
always happen in practice!

Useful resources and sources for further study

Berry, R. (2004) The Research Project: How to Write it, McMillan, K.M. and Weyers, J.D.B. (2014) How to com-
5th edn. Routledge, London. plete a successful Research Report. Pearson Education,
London.
Davis, M. and Davis K.J. (2012) Scientific Papers and
Presentations: Navigating Scientific Communication in Matthews, J.R., Bowen, J.M. and Matthews, R.W. (2007)
Today’s World, 3rd edn. Academic Press, London. Successful Science Writing: A Step-by-step Guide for the
Biological and Medical Sciences, 3rd edn. Cambridge
Day, R.A. and Gastel, B. (2012) How to Write and Publish University Press, Cambridege.
a Scientific Paper, 7th edn. Cambridge University Press, McCormac, C., Davis, J., Papakonstantinou, P. and Ward,
Cambridge. N. (2012) Research Project Success. The essential guide
Hofmann, A.H (2014) scientific writing and communi- for science and engineering students. Royal Society of
cation: papers, proposals and presentations, 2nd edn. Chemistry, Cambridge.
Oxford university Press, Oxford. Valiela, I. (2009) Doing Science: Design, Analysis and
Lobban, C.S. and Schefter, M. (1992) Successful Lab Communication of Scientific Research, 2nd edn. Oxford
University Press, Oxford.
Reports: A Manual for Science Students. Cambridge
[Covers scientific communication, graphical presenta-
University Press, Cambridge.
tions and aspects of statistics.]
Luey, B. (2009) Handbook for Academic Authors, 5th edn. Vitae available via www.vitae.ac.uk (last accessed
Cambridge University Press, Cambridge. 21/03/16).
Study exercises
70.1 Write a formal ‘Experimental’ section. Adopting to comment on your description, to identify what
the style of a research paper (i.e. past tense, all is missing or unclear.
relevant detail reported such that a competent
70.3 Write an abstract for a paper. Pair up with a col-
colleague could repeat your work), write out the
league. Each of you should independently choose
Experimental for a practical you have recently
a different research paper in a current journal.
carried out. Ask a colleague or tutor to comment
Copy the paper, but mask over the abstract sec-
on what you have written.
tion, having first counted the words used. Swap
70.2 Describe a set of results in words. Again adopt- papers. Now, working to the same number of
ing the style of a research paper, write a par- words as in the original, read the paper and pro-
agraph describing the results contained in a vide an abstract of its contents. Then compare this
particular table or graph. Ask a colleague or tutor with the real abstract. Compare your abstracts.
Answers to these study exercises are available at www.pearsoned.co.uk/practicalskills

71 Writing literature surveys and reviews
8 (5%) 1 (5%) The literature survey or review (sometimes termed a ‘dissertation’) is a special-
7 (10%) ised form of essay which summarises and reviews the evidence and concepts
concerning a particular area of research.
6 (5%)
5 (5%) KEY POINT A literature review should not be a simple recita-

2 (45%) tion of facts. The best reviews are those which analyse informa-
tion rather than simply describe it.
4 (10%)
Organising your time

3 (15%)
Figure 71.1 illustrates how you might divide up your time for writing a litera-
Fig. 71.1 Pie chart showing how you might ture survey. There are many subdivisions in this chart because of the size of the
allocate time for a literature survey: task: in general, for lengthy tasks, it is best to divide up the work into managea-
1. select a topic ble chunks. Note also that proportionately less time is allocated to writing itself
2. scan the literature than with an essay. In a literature survey, make sure that you spend adequate
3. plan the review time on research and revision.
4. write first draft
5. leave to settle
Selecting a topic
6. prepare a structured review of text
7. write final draft You may have no choice in the topic to be covered, but if you do, carry out your
8. produce top copy. selection as a three-stage process:
1. Identify a broad subject area that interests you.
2. Find and read relevant literature in that area. Try to gain a broad
impression of the field from books and general review articles. Discuss
Creating a glossary – one barrier to your ideas with your supervisor.
developing an understanding of a new
3. Select a relevant and concise title. The wording should be considered
topic is the jargon used. To overcome
very carefully as it will define the content expected by the reader. A narrow
this, create your own glossary. You
subject area will cut down on the amount of literature you will be expected
may wish to cross-reference a range
to review, but will also restrict the scope of the conclusions you can make
of sources to ensure the definitions are
(and vice versa for a wide subject area).
reliable and context-specific. Remem-
ber to note your sources in case you
wish to use the definition within your Scanning the literature and organising your references
review.
You will need to carry out a thorough investigation of the literature before you
start to write. The key issues are as follows:
●● Make a start with relevant literature. Seek help from your supervisor,
who may be willing to supply a few key papers to get you started. Hints on
expanding your collection of references are given on p. 379.
Using index cards – these can help
you organise large numbers of ref- ●● Assess the relevance and value of each article. This is the essence of writ-
erences. Write key points and author ing a review, but it is difficult unless you already have a good understanding
information on each card – this helps of the field. Try reading earlier reviews in your area and discussing the topic
when considering where the reference with your supervisor or other academic staff.
fits into the literature. Arrange the cards
in subject piles, eliminating irrelevant
●● Clarify your thoughts. One approach is to use the SPSER method
ones. Order the cards in the sequence
(Box 71.1). Alternatively, by subdividing the main topic and assigning your
in which you wish to write.
references to these smaller subject areas may help you may gain a better
overview of the literature.

Writing literature surveys and reviews
Box 71.1 How to analyse a topic using the SPSER approach
This method is useful when trying to get to grips with Introduction. In some cases this will be clear from the
a new and complex subject. The approach helps you to title of the exercise you have been given.
‘deconstruct’ the topic and involves considering the task 3. Solutions. Outline possible answers to questions
in five discrete stages, given the acronym SPSER. You can raised, or ways of tackling the problem. These will
note down your thoughts using a table or the ‘pattern probably arise from your reading of papers, reviews
notes’ (p. 542) or ‘mind map’ (p. 543) approaches, ready and other sources.
for converting into more formal text.
4. Evaluation. Note positive or negative features of each
1. Situation. Briefly outline the context of the topic and the solution and provide evidence to support your view-
history of its development. For some complex topics point. A tabular approach might be useful, listing pos-
you may need to refer to broader introductory texts or itives on one side of a vertical line and negatives on
websites to build a foundation for your understanding. the other.
2. Problem. State the essential issues, questions or 5. Recommendation. Arrive at a conclusion by decid-
problems that researchers or others are tackling. In a ing what might/should happen next, outlining your
research paper, this might be stated succinctly in the reasoning.
Abstract, but you might also find it at the end of the
Deciding on structure and content

The general structure and content of a literature survey are described below.
The Chemical Society Reviews series (available in most university libraries)
provides good examples of appropriate style for reviews of the chemical sciences.
Introduction
Defining terms – the introduction is a The introduction should give the general background to the research area, con-
good place to explain the meaning of centrating on its development and importance. You should also make a state-
the key terms used in your survey or ment about the scope of your survey; as well as defining the subject matter to
review. be discussed, you may wish to restrict the period being considered.
Main body of text
The review itself should discuss the published work in the selected field and
may be subdivided into appropriate sections. Within each portion of a review,
the approach is usually chronological, with appropriate linking phrases (e.g.
‘Following on from this, . . .’; ‘Meanwhile, Bloggs (2014) tackled the problem
from a different angle . . .’). However, a good review is much more than a
chronological list of work done. It should:
●● allow the reader to obtain an overall view of the current state of the
research area, identifying the key areas where knowledge is advancing;
Balancing opposing views – even if ●● show how techniques are developing and discuss the benefits and disad-
you favour one side of a disagreement vantages of using particular organisms or experimental systems;
in the literature, your review should ●● assess the relative worth of different types of evidence – this is the most
provide a balanced and fair descrip- important aspect (see Chapter 42). Do not be intimidated from taking a
tion of all the published views of the critical approach as the conclusions you may read in the primary literature
topic. Having done this, if you do wish are not always correct;
to state a preference, give reasons for
●● indicate where there is conflict in findings or theories, suggesting, if pos-
your opinion.
sible, which side has the stronger case;
●● indicate gaps in current knowledge.
You do not need to wait until you have read all the sources available to you
before starting to write the main body. Word processors allow you to modify
and move pieces of text at any point and it will be useful to write paragraphs

Writing literature surveys and reviews
about key sources, or groups of related papers, as you read them. Try to create
a general plan for your review as soon as possible. Place your draft sections
of text under an appropriate set of subheadings that reflects your plan, but be
prepared to rearrange these and re-title or re-order sections as you proceed. Not
only will working in this way help to clarify your thoughts, but it may help you
avoid a last-minute rush of writing near to the submission date.
Conclusions
The conclusions should draw together the threads of the preceding parts and
point the way forward, perhaps listing areas of ignorance or where the applica-
Making citations – a review of litera- tion of new techniques may lead to advances.
ture poses stylistic problems because References, etc.
of the need to cite large numbers of
papers; in the Chemical Society Reviews
The references or literature Cited section should provide full details of all papers
series this is overcome by using num-
referred to in the text (see p. 383). The regulations for your department may
bered references (see p. 382).
also specify a format and position for the title page, list of contents, acknowl-
edgements, etc.
Source for further study

Holst, B. (2015) Scientific paper writing: A survival guide. McMillan, K.M. and Weyers, J.D.B. (2013) How to Write
Create Space Independent, London. Dissertations and Project Reports, 2nd edn. Prentice
McCormac, C., Davis, J., Papakonstantinou, P. and Hall, Harlow.
Ward, N. (2012) Research Project Success. The essential Rudner, L.M. and Schafer, W.D. (1999) How to write
guide for science an engineering students. Royal Society a scholarly research report. Practical Assessment,
of Chemistry, Cambridge. Research & Evaluation, 6 (13). Available: http://
McMillan, K.M. and Weyers, J.D.B. (2013) The Smarter Study pareonline.net/getvn.asp?v=6&n=13
Skills Companion. 3rd edn. Pearson Education, Harlow. Last accessed: 01/09/15.
Study exercises
71.1 Summarise the main differences between a and relevant to the topic? List five of these, using
review and a scientific paper. From the many sub- the proper conventions for citing articles in a
ject areas in the Chemical Society Reviews series reference list (see Chapter 9). Note that each of
(find via your library’s periodical indexing sys- these papers will also cite other articles, always
tem), pick one that matches your subject interests, going back in time. Now using the Science Cita-
and within this find a review that seems relevant tion Index or a similar system (e.g. the Web of
or interesting. Read the review and write down Science website at http://wos.mimas.ac.uk or
five ways in which the writing style and content Google Scholar at http://scholar.google.com/),
differ from those seen in primary scientific papers. work forward and find out who has cited your
selected article in the time since its publication.
71.2 Gather a collection of primary sources for a topic.
Again, list the five most important articles found.
From the journal section of the library, select an
interesting scientific paper published about 5–10 71.3 Write a synopsis of a review. Using one of the
years ago. First, work back from the references Annual Review series as a source, allow yourself
cited by that paper: can you identify from the text just five single-sentence bullet points to summa-
or the article titles which are the most important rise the key points reported in a particular review.

Answers to study exercises
We have attempted to provide an ‘answer’ to all of the study exercises. Where the question is open-ended and no ‘correct’ answer
can be given, we have provided a Tip which should either help with your general approach, indicating which resources are worth
consulting, or provide a pointer to relevant material within the book. Where a non-numerical question has a ‘correct’ answer, we have
provided a model text-based answer. If a calculation is involved, we have shown the steps involved and have indicated the correct
answer underlined in bold.
1 Essentials of practical work (c) Sodium hydroxide is highly corrosive, poisonous and an
1.1 Possible reasons why practical work might be of value in a irritant. Protective gloves and safety glasses must be worn
university course include: when making solutions using solid NaOH. Contact with
●● practicals reinforce lecture material, in agreement with moisture generates considerable heat so it should always
the adage: ‘I hear and I forget, I see and I remember, I do be added in small amounts to a large volume of water.
and I understand’;
●● practical procedures provide students with an opportunity 3 Making measurements
to develop their manual skills and laboratory competences; 3.1 (a) quantitative, discontinuous, ratio
●● investigative procedures and problem-solving practical (b) quantitative, continuous, ratio
exercises enable students to improve their skills in experi- (c) qualitative, discontinuous, nominal
mental design and application of scientific method; (d) quantitative, discontinuous, ratio
●● practicals enable students to develop their abilities to observe (e) qualitative, discontinuous, nominal
and measure chemical systems, and to record the outcome; (f) qualitative, discontinuous, ordinal
●● the results of practical procedures give students an oppor- 3.2 The results indicate that balance A has a bias of + 0.05 g
tunity to develop skills in reporting and presenting ‘exper- across the weighing range, while balance B has a consistent
iments’ in a written format. bias of about +0.04%. Both balances are precise, but not ac-
1.2 Tip: Possible items for aqueous recrystallisation: Bunsen burn- curate.
er, tripod and gauze; two 250 mL conical flasks; anti-bumping 3.3 Tip: Use a spreadsheet to lay out the table once you have de-
granules or boiling stick or glass rod; small watch glass, clock cided what it needs to contain and what statistics you might
glass; stemless filter funnel: fluted filter paper; charcoal, two- wish to calculate. You can use spreadsheet functions to help
dp balance; spatula; Büchner flask and funnel; rubber collar; you calculate some of the statistics.
filter paper; ice/water bath; water suction pump; drying oven.
1.3 (a) 40; 4 SI units and their use
(b) 44.266 66, expressed to 4 significant figures = 44.27; 4.1 Tip: Box 4.1 gives procedures for interconversion of units.
(c) 0.019 531 25, expressed to 3 significant figures = 0.0195; (a) Rearrange the equation and add units:
(d) 1.6 * 105; PV kg m-1 s-2 * m3
(e) 0.0313, expressed to 3 decimal places = 0.031. R = = = kg m2 s-2 mol-1K-1
nT mol * K
2 Health and Safety = J mol-1K-1
2.1 (a) p. 6; (b) Rearrange the equation and add units:
(b) Fig. 2.1 (p. 7); A 1
(c) Fig. 2.3 (p. 10); e = = -1
= mol-1 L * (10-2 m) -1
c * l mol L * cm
(d) Fig. 2.3 (p. 10).
2.2 Tip: The locations of each item will vary, according to the = mol-1 * 10-3 m3 * 102 m-1 = mol-1 10-1 m2
layout of your chosen laboratory. You should ask someone in (c) Square the equation and rearrange:
charge (e.g. lecturer technician, demonstrator or laboratory k = v 2 * m * 4p2 = s-2 * kg = kg s ∙ 2
manager) if you are unable to find any of the items listed.
2.3 Tip: Use either your department’s chemical hazard informa-
5 Scientific method and design of experiments
tion system or an appropriate reference text (e.g. Merck Index
5.1 A possible answer (one of many) is given in the figure:
or Bretherick) or the online MSDS available.
(a) Ethanol is highly flammable, with a b.pt. 78 °C. Eye
protection must be worn. Use in fume cupboard with no W Z Y X Z W X Y W Y X Z
naked flames. Water bath or spark proof electric mantle
X W Z Y W Z Y X Y Z W X
as heat source and reflux apparatus (pp. 89–91) to prevent
loss of solvent during heating. Y X W Z Y X W Z Z X Y W
(b) Sodium oxalate is highly poisonous. Eye protection and Z Y X W X Y Z W X W Z Y
gloves must be worn for all operations involving oxalates.
Dispose of residual solutions and gloves appropriately,
and wash all glassware thoroughly with water. A possible layout of treatments in the glasshouse experiment.
Z01 Practical Skills in Chemistry 39920.indd 609 12/05/2017 15:52

The Latin square design ensures that treatments appear with 9 Basic laboratory procedures I
equal probability in each row and column; hence, if there are 9.1 Tip: Box 9.1 and Box 9.2 give advice on preparing solutions:
gradients of potentially confounding variables, such as tem- (a) volume = 0.25 L: use eqn [9.1] 0.05 = amount (mol) ,
perature and light in this case, their effects are spread evenly 0.25, thus amount (mol) = 0.05 * 0.25 = 0.0125 mol;
among treatments. Mr for NaCl = 58.44 g mol-1; convert mol to g by
5.2 Tip: For Microsoft Excel, try the formula = multiplying = 58.44 * 0.0125 = 0.7305 g.
INT(5*(RAND()) + 1). (b) volume = 0.1 L: use eqn [9.1] 0.02 = amount (mol) ,
0.1, thus amount (mol) = 0.02 * 0.1 = 0.002 mol; Mr
6 Making notes on practical work
for KIO3 = 214.00 g mol-1; convert mol to g by
6.1 Accurate weights (to 4 decimal places) of original sample;
multiplying = 214.00 * 0.002 = 0.4280 g.
precipitated sample; gooch crucible (before and after).
(c) volume = 0.05 L: use eqn [9.1] 0.05 = amount (mol) ,
●● Drying temperature and duration.
0.05, thus amount (mol) = 0.05 * 0.05 = 0.0025 mol;
6.2 As well as information in 6.1 also perform calculation as
Mr for Na 2S2O3.5H2O = 248.18 g mol-1; convert mol to
shown in Box 22.1.
g by multiplying = 248.18 * 0.0025 = 0.6205 g.
7 Project work (d) volume = 0.25 L: use eqn [9.1] 0.1 = amount (mol) ,
7.1 Tips: 0.25, thus amount (mol) = 0.1 * 0.25 = 0.025 mol; Mr
●● Divide the time available for your project into relevant for CuSO4.5H2O = 249.60 g mol-1; convert mol to g by
parts concerned with different aspects of your work (note multiplying = 249.60 * 0.025 = 6.2400 g.
some of these could run in parallel such as experimentation (e) Note each molecule contains two potassium ions,
and research for your Introduction). therefore you need half the number of moles of
●● A Gantt chart might be an appropriate way of presenting K2SO4 i.e. 0.025 mol; volume = 0.1 L: use eqn.
this (see, e.g., http://associate.comk/gantt). [9.1] 0.025 = amount (mol) , 0.1, thus amount
●● Set yourself realistic target dates for completion of each (mol) = 0.025 * 0.1 = 0.0025 mol; Mr for K2SO4 =
section. 172.47 g mol-1; convert mol to g by multiplying = 172.47
●● Identify important ‘sticking points’ (e.g. time taken for *0.0025 = 4.3118 g.
delivery of chemicals and other supplies) and work round 9.2 Tip: For all these calculations you can use [C1]V1 = [C2]V2 as
there (e.g. by doing library work). outlined in Box 9.2.
●● Make sure you allow some flexibility (say 10%) to allow (a) 0.4 * 1.0 = [C2] * 10; therefore [C2] = 0.04 M;
for slippage or unforeseen problems). (b) 0.1 * V1 = 0.02 * 500; therefore V1 = 100.00 mL;
7.2 Tip: Even if you prefer to write up your work on paper first, (c) [C1] * 10.00 = 0.001 * 250; therefore [C1] = 0.025 M.
create appropriate computer files soon afterwards. Devise a (d) 0.02 * 5.00 = 0.001 * V2; therefore V2 = 100.00 mL.
simple and logical system for naming the files.
7.3 Tip: Remember that a word processor or a spreadsheet may 10 Basic laboratory procedures II
have sufficient functions for your database needs for storing 10.1 (a) Either (i) conical flask or beaker, stemmed funnel and
reference details (see p. 436). fluted filter paper or (ii) Büchner flask and funnel, rubber
collar, filter paper, water vacuum.
8 Working with liquids (b) Büchner flask and funnel, rubber collar, filter paper, water
8.1 (a) Use measuring cylinders (e.g. measure out 700 mL of vacuum pump.
ethanol using a 1000 mL measuring cylinder and measure (c) Conical flask, stemless funnel and fluted filter paper, hot
out 300 mL of water using a 500 mL measuring cylinder: plate (to keep filtrate and funnel and filter paper hot to
mix together in a large conical flask or beaker with a ca- prevent crystallisation.
pacity of 7 1000 mL). (d) Hot plate or Bunsen burner, tripod and gauze, conical
(b) Use a pipette (25.00 mL) with three valve pipette filler. flask, watch glass (to prevent rapid evaporation of water,
(Fig. 8.2). Using the procedure described in Chapter 24 anti-bumping precautions.
(p. 195) to transfer conc HCl (25.00 mL) to water (e) Reflux set up (p. 89–91), oil bath or hot plate, anti-bumping
(100 mL by measuring cylinder) in a beaker (250 mL). precautions.
Stir with a glass rode to ensure complete mixing. (f) Ice–CaCl2.6H2O bath (see Table 10.2), plastic bowl or
(c) Use a pipettor set to deliver 10 mL to transfer the pro- beaker, clamp and stand, thermometer.
panone into the volumetric flask containing distilled water
(∙ 20 mL). This prevents loss of propanone by evapora- 11 Principles of solution chemistry
tion. Make up to the mark with distilled water using a 11.1 Tip: Box 9.2 and Box 10.1 give useful procedures and exam-
Pasteur pipette for the final adjustment. Stopper the flask ple calculations for molar concentrations;
and invert at least eight times to ensure complete mixing. (a) 1000 mL of 1.0 M NaCl = 1.0 mol L-1 thus need 1 mol;
(d) Use a pipette (25.00 mL) to transfer the NaOH solution to Mr NaCl = 58.44 g mol-1 - weigh out 58.4400 g.
a conical flask (250 mL) and deliver the H2SO4 solution (b) 1000 mL of 0.2004 M KMnO4 = 0.2004 mol L-1 thus
from a burette. (See Chapter 24) need 0.2004/4 mol = 0.0501 mol; Mr for KMnO4 =
8.2 Tip: Box 8.1 (p. 66) gives stepwise details of how to use a 158.04 g mol-1 - weight out 158.04 * 0.0501 =
pipettor. 7.9178 g.
610 Answers to study exercises

(c) 5% of 400 = 20; weigh out 20.0000 g of NaOH and dis- Balance the central element, balance oxygen by adding
solve in 400 mL of water. water and balance hydrogen by adding H +:
(d) 10% of 300 = 30; weigh out 30.0000 g of KNO3 and 12H + + 2IO3- S I2 + 6H2O
dissolve in 270 g (270 mL since density = 1 g mL-1) of
2I - S I2
water.
11.2 Tip: Make stepwise changes and always show your working Balance the charge on each side of the equations by add-
out in any written report or assessment in calculations involving electrons and then balance the electrons since number
ing the interconversion of concentrations. of electrons gained equals number of electrons lost:
(a) 4 , 40 (convert g L-1 to mol L-1) = 0.1000 mol L- 1. 10e + 12H + + 2IO3- S I2 + 6H2O
(b) 0.1 * 294.19 (convert mol L-1 to g L-1) = 29.4190 g L-1.
10I - S 5I2 + 10e
(c) 5 * 10 (convert % to mL of ethanol in 1000 mL =
50 mL; 50 * 07892 (convert mL to g) = 39.4600 g; Add the partial ionic equations to give the overall balanced
39.4600 , 46 (convert g to mol) = 0.8578 mol L∙ 1. ionic equation:
(d) 150 , 1000 (convert mmol to mol) = 0.1500 mol L-1: 10I - + 12H + + 2IO3- S 6I2 + 6H2O
0.1500 * 180 (convert mol to g) = 27.0000 g L-1 =
Divide by 2:
27.0000%.
11.3 Tip: Box 11.4 and Box 11.5 give procedures for balancing 5I - + 6H + + IO3- S 3I2 + 3H2O
redox equations. (d) Identify the partial equations for reduction and oxidation
(a) Identify the partial equations for reduction and oxidation using oxidation numbers:
using oxidation numbers:
(COOH)2 S CO2 and MnO4- S Mn2+
Cr2O2-
7 S Cr
3+
and Fe2+ S Fe3+
Balance the central element, balance oxygen by adding
Balance the central element, balance oxygen by adding water and balance hydrogen by adding H -:
water and balance hydrogen by adding H +
(COOH)2 S 2CO2 + 2H +
14 H + + Cr2O2-
7 S 2Cr
3+
+ 7H2O
2+ S 3+
8H + + MnO4- S Mn2+ + 4H2O
Fe Fe
Balance the charge on each side of the equations by add-
Balance the charge on each side of the equations by adding electrons and then balance the electrons since number
ing electrons and then balance the electrons, since number of electrons gained equals number of electrons lost:
of electrons gained equals number of electrons lost:
5(COOH)2 S 10CO2 + 10H + + 10e
6e + 14H + + Cr2O2-
7 S 2Cr
3+
+ 7H2O
2+ S 3+
10e + 16H + + 2 MnO4- S 2 Mn2+ + 8H2O
6Fe 6Fe + 6e
Add the partial ionic equations to give the overall balanced
Add the partial ionic equations to give the overall balanced ionic equation:
ionic equation:
5(COOH)2 + 6H + + 2MnO4-
14H + + 6Fe2+ + Cr2O2-
7 S 2Cr
3+
+ 6Fe3+ + 7H2O
S 2Mn2+ + 10CO2 + 8H2O
(b) Identify the partial equations for reduction and oxidation
using oxidation numbers: 12 pH and buffer solutions
S2O2- 2-
3 S S4O6 and I2 S I
- 12.1 Tip: Use eqn [12.5] to interconvert between pH and [H +].
(a) 7.4 = - log10[H +], therefore [H +] = 3.98 : 10∙8
Balance the central element, balance oxygen by adding
mol L∙ 1.
water and balance hydrogen by adding H +:
(b) 4.1 = - log10[H +], therefore [H +] = 7.94 : 10∙5
2S2O2- 2-
3 S S4O6 mol L∙ 1.
-
I2 S 2I (c) pH = -log10[2 * 10-5], therefore pH = 4.70.
Balance the charge on each side of the equations by add- (d) pH = -log10[2 * 10-12.5], therefore pH ∙ 12.50.
ing electrons and then balance the electrons since number 12.2 Tip: Rearrange eqn [12.7] to give log10[A-]/[HA] =
of electrons gained equals number of electrons lost: pH - pKa
(a) log10 [A-]/[HA] = 3.8 - 4.8 = -1, so [A-]/[HA] =
2S2O2- 2-
3 S S4O6 + 2e 0.1 (1 to 10).
2e + I2 S 2I - (b) log10[A-]/[HA] = 9.5 - 9.2 = 0.3 so [A-]/[HA] =
Add the partial ionic equations to give the overall balanced 1.995 (approximately 2 to 1).
ionic equation: (c) log10[A-]/[HA] = 8.1 - 7.5 = 0.6 so [A-]/[HA] =
-
3.98 (approximately 4 to 1).
2S2O2- 2-
3 + I2 u S4O6 + 2I
(c) Identify the partial equations for reduction and oxidation 13 Melting points
using oxidation numbers: 13.1 (1) The product is impure and needs to be purified by recrys-
IO3- S I2 and I - S I2 tallisation.
(2) The compound is NOT N-phenylethanamide.

(3) The thermometer used should be calibrated. 18 Evaporation

13.2 (1) The compound is NOT benzoic acid. 18.1 (a) Because of small volume and boiling point considera-
(2) You have heated the sample too rapidly and overshot the tions can use open flask on water bath.
actual m.pt. (b) Large volume implies use of a ‘rovap’.
(3) The thermometer used should have been calibrated. (c) Very small volume implies use of gas ‘blow-down’ with
13.3 Your product IS 4-nitroaniline, since the mixed melting point little or no heating.
is undepressed.
19 Inert atmosphere methods
19.1 (a) Ethoxyethane: test for peroxides using acidified 10%
14 Recrystallisation
(w/v) aqueous potassium iodide. If positive, shake the
14.1 Your list should include the following: hot water bath or
ethoxyethane with 1/5 volume 5% (w/v) aqueous sodi-
hot plate, two conical flasks, boiling sticks/glass rods/anti-
um bisulphite until test for peroxides is negative. Sepa-
bumping granules, watch glass, decolorising charcoal, stem-
rate the ethoxyethane. Dry by standing over anhydrous
less filter funnel, fluted filter paper, ice-water bath, Büchner
MgSO4 overnight, filter, and stand over sodium wire for
funnel and flask, filter paper to fit, water vacuum system, two
two days (do not seal the flask/bottle since H2 is evolved.
clock glasses, drying oven, balance (2 dp).
Decant the ethoxyethane through a fluted filter system
and distil (b.pt. 35 °C) in clean, dry apparatus. Do not
15 Solvent extraction distil to dryness.
15.1 (1) Clamp on stem of funnel - unless the funnel is absolute- (b) Tetrahydrofuran (THF): test for peroxides using acidified
ly vertical, the mass of the liquid will cause the funnel to 10% (w/v) aqueous potassium iodide. If positive, shake
tip sideways and the stem may snap. the THF with 1/5 volume 5% (w/v) aqueous sodium
(2) The tap is open. bisulphite until test for peroxides is negative. Dry by
(3) There is no ‘safety’ beaker. standing over KOH pellets overnight, decant off the THF
(4) There is no stemmed funnel to aid in filling the separatory and then reflux the THF over sodium and a little benzo-
funnel. Liquid will evaporate from the joint, resulting in phenone until a deep purple colour of sodium benzophe-
a poor fit of the stopper and hence spillage when the sep- none ketyl develops. Distil (b.pt. 64–66 °C) THF from
aratory funnel is inverted. the sodium benzophenone ketyl and metallic sodium. Do
not distil to dryness. If only a small volume of THF is
16 Distillation required, it can be dried by passing it through a column
16.1 Tip: Remember that this is only an approximate estimation: of Activity 1 alumina.
(a) 760 to 190 is equivalent to 2 * 10 = 20 °C, therefore (c) Hexane: Dry by standing over anhydrous MgSO4 over-
b.pt. ? 320 ∙C. night, filter off the MgSO4 and distil (b.pt. 67–71 °C),
(b) 760 to 100 is equivalent to 3 * 10 = 30 °C, therefore rejecting the fore-run, which contains water. Hexane is
b.pt. ? 310 ∙C. poisonous.
(c) 760 to 20 is equivalent to 5 * 10 = 50 °C, therefore
b.pt. ? 290 ∙C. 20 Combinatorial chemistry
(d) 760 to 1.0 is equivalent to 9 * 10 = 90 °C, therefore 20.1 20 * 20 * 20 = 8000.
b.pt. ? 250 ∙C. 20.2 (12 * 5 * 8)/96 = 5 plates.
(e) 760 to 0.1 is equivalent to 12 * 10 = 120 °C, therefore
b.pt. ? 220 ∙C. 21 Qualitative techniques for inorganic chemistry
16.2 (a) 275 °C; 21.1 Tap water contains a selection of inorganic ions which will
(b) 260 °C; give ‘false-positive’ results and therefore an incorrect analy-
(c) 222 °C; sis.
(d) 135 °C; 21.2 The first test indicates the presence of carbonate (CO2- 3 ) or
(e) 110 °C. bicarbonate (HCO3-) ions. The second test indicates the pres-
ence of barium (Ba2+) or copper (Cu2+) ions. Copper and bar-
ium bicarbonates do not exist as solids and copper carbonate
17 Reflux only as the basic carbonate CuCO3.Cu(OH)2 in which the OH
17.1 Hazard Assessment: cyclohexene - b.pt. = 58.8 °C, flam- group is not detected by the tests. The unknown solid could be
mable, harmful by absorption (skin and lungs), use in fume CuCO3.Cu(OH)2 or BaCO3, but since the former is blue and
cupboard; bromine - b.pt. = 83 °C, extremely harmful. Cor- the latter is white an easy distinction can be made.
rosive and toxic liquid and vapour, wear gloves and use in fume
cupboard; (E)-1,2 dibromocyclohexane - b.pt. = 145°C 22 Gravimetry
at 100mm Hg, flammable, harmful by absorption (skin and 22.1 Tip: Work in moles and then convert to grams. For % (w/w)
lungs), use in fume cupboard. Apparatus required is illus- need g of NaCl in 100 g of sea water. Assume the density of
trated in Fig 17.6(a). Requires use of pressure equalising sea water is 1.00 g mL-1:
dropping funnel to prevent escape of bromine vapour during 1 mol of AgCI K 1 mol of NaCl
addition. 0.8329 K 0.8329/143.32 mol = 0.005812 mol of AgCl

Therefore 10.00 mL of seawater contains 0.005812 mol 1000 mL of 0.1 M H2SO4 K 1.0 * 0.1 mol of Na 2CO3
of NaCl: 1.0 * 0.1
1.00 mL of 0.1 M H2SO4 K mol of Na 2CO3
0.005812 mol of NaCl = 58.5 * 0.005812 1000
= 0.3400 g of NaCl 1.0 * 0.1 * 26.00
26.00 mL of 0.1 M H2SO4 K mol
Therefore 100 mL K 100 g of sea water contain 3.4000 g 1000
of NaCl = 3.4% (w/w). of Na 2CO3 = 0.0026 mol Na 2CO3
22.2 Tap water contains Cl- ions which will nullify the determina- Therefore 25.00 mL of Na 2CO3 solution contain 0.0026 mol
tion. of Na 2CO3
Therefore 1000 mL of Na 2CO3 solution contain
23 Molecular Formulae
0.0026 * 1000
23.1 (a)
C = 64.86/12.01 = 5.4, H = 13.51/1.008 = 13.40, = 0.1040 mol L-1 Na2CO3
25.00
O = 21.62/16 = 1.35. Divide by smallest number:
C = 5.4/1.351 = 4, H = 13.40/1.351 = 9.925, O = (ii) First the equation:
1.351/1.351 = 1. Rounding to whole numbers gives EF 2NaOH + H2SO4 S Na 2SO4 + 2H2O
as C4H10O. Mr is 74, thus EF = MF as C4H10O.
1 mol H2SO4 reacts with 2 mol NaOH
(b) Data total is not 100 %, therefore oxygen present.
Subtract sum from 100 to give O as 35.56%. Then 1000 mL of 1.0 M H2SO4 K 2.0 mol of NaOH
C = 53.33/12.01 = 4.40, H = 11.11/1.008 = 11.02, 1000 mL of 0.05 M H2SO4 K 2.0 * 0.05 mol of NaOH
O = 35.56/16 = 2.22. Divide by smallest number:
C = 4.40/2.22 = 1.98, H = 11.02/2.22 = 4.96, O = 1.00 mL of 0.05 M H2SO4
2.22/2.22 = 1. Rounding up to whole numbers gives EF 2.0 * 0.05
as C2H5O. Mr is 90 therefore MF = 2 x EF as C4H10O2. K mol of NaOH
1000
(c) Data total is not 100%, therefore oxygen present.
24.00 mL of 0.05 M H2SO4
Subtract sum from 100 to give O as 36.36%. Then
C = 54.55/12.01 = 4.54, H = 9.09/1.008 = 9.02, 2.0 * 0.05 * 24.00
K mol of NaOH
O = 36.36/16 = 2.27. Divide by smallest number: 1000
C = 4.54/2.27 = 2, H = 9.02/2.27 = 3.97, = 0.0024 mol NaOH
O = 2.27/2.27 = 1. Rounding up to whole numbers Therefore 25.00 mL of NaOH solution contains 0.0024 mol
gives EF as C2H4O. Mr is 88 therefore MF = 2 x EF as of NaOH
C4H8O2. Therefore 1000 mL of NaOH solution contain
(d) Data total is not 100%, therefore oxygen present.
0.0024 * 1000
Subtract sum from 100 to give O as 12.50%. Then = 0.0960 mol L-1 NaOH
C = 56.25/12.01 = 4.48, H = 3.90/1.008 = 3.87, 25.00
Cl = 27.34/35.5 = 0.78, O = 12.5/16 = 0.78. 24.2 12 drops = 1.00 mL, therefore 1 drop = 0.08333 mL; there-
Divide by smallest number C = 4.68/0.78 = 6, fore 8 drops = 0.6667 mL; therefore total titre = 25.20 +
H = 3.87/0.78 = 4.96, Cl = 0.78/0.78 = 1, 0.6667 = 25.87 mL.
O = 0.78/0.78 = 1. Rounding up to whole numbers
gives EF as C6H5ClO. Mr is 128 thus EF = MF as 25 Acid–base titrations
C6H5CIO.
2HClaq + Na 2CO3.xH2Oaq S 2NaClaq + CO2g
[(6 + 2) - (5 - 1 + 0)]
23.2 (a) For C3H5N: DBE = = 2. + (x + 2)H2Oaq
2
[(16 + 2) - (14 - 0 + 0)] 2 mol of HCl K 1 mol of Na 2CO3.xH2O
(b) For C8H14O4: DBE = = 2.
2 2 * 1000 mL of HC1 (1.0 M) K 1 mol of Na 2CO3.xH2O
[(10 + 2) - (11 - 1 + 0)]
(c) For C5H11N: DBE = = 1. 2 * 1000 mL of HC1 (0.1 M) K 0.1 * 1 mol of
2
Na 2CO3.xH2O
[(20 + 2) - (8 - 0 + 0)]
(d) For C10H8: DBE = = 7. 25.4 * 0.1 * 1
2 25.4 mL of HCl (0.1 M) K mol of
2 * 1000
24 Procedures in volumetric analysis
24.1 First the equation: Na 2CO3.xH2O
Na 2CO3 + H2SO4 S Na 2SO4 + H2O + CO2 K 0.00127 mol of Na 2CO3.xH2O
1 mol H2SO4 reacts with 1 mol Na 2CO3 Therefore 25.00 mL of Na 2CO3.xH2O contains 0.00127 mol
We know the concentration and volume of the H2SO4 and of Na 2CO3.xH2O
the volume of Na 2CO3 solution. We need to find the num- Therefore 250.00 mL of Na 2CO3.xH2O contains 0.0127 mol
ber of moles of Na 2CO3 in 1.0 L = 1000 mL. of Na 2CO3.xH2O
Therefore 0.0127 mol of Na 2CO3.xH2O = 3.6284 g of
1000 mL of 1.0 M H2SO4 K 1.0 mol of Na 2CO3 Na 2CO3.xH2O

3.6284 * 1 g 0.000375 * 2000

Therefore 1.0 mol of Na 2CO3.xH2O = of = 0.03 mol of Ca2+ ions
0.0127 mol 25.00
Na 2CO3.xH2O and in 2000 mL of diluted urine solution there must be
= 285.7 g mol-1 0.000825 * 2000
= 0.066 mol of Mg 2+ ions
= Mr 25.00
Na 2CO3.xH2O = 46 + 12 + 48 + (x * 18) = 285.7 Amount of Mg 2+ in diluted urine solution = 0.066 *
24.312 * 1000 = 1605 mg
Therefore 18x = 285.7 - 106 Amount of Ca2+ in diluted urine solution = 0.03 * 40.08 *
Therefore x = 9.983. But x must be a whole number 1000 = 1202 mg
Therefore formula is Na2CO3.10H2O
26 Complexometric titrations 27 Redox titrations

26.1 Tip: Do not be flustered by the apparent complexity of the 27.1 Using the principles outlined in Chapter 11, identify the par-
procedure and remember the following points: tial ionic equations and balance the full redox equation:
●● EDTA reacts with metal ions in a 1:1 ratio.
●● The first titration gives the total metal (Mg
2+
and Ca2+ ) 2H2O2 S 2H2O + O2
ion concentration/amount in 25.00 mL of diluted urine MnO4- S Mn2+
solution. 2 MnO4- + 2H2O2 + 6H + S 2 Mn2+ + 8H2O + 5O2
●● The second titration is known as a ‘back titration’ in which
Therefore
the amount of Ca2+ ion is measured by adding a known
excess of EDTA and then titrating the excess EDTA with a 2 moles of KMnO4 K 5 moles of H2O2
metal ion solution, in this case Mg 2+. This tells us how much 2 * 1000 mL of 1.0 M KMnO4 K 5 moles of H2O2
Ca2+ there is in 50.00 mL of diluted urine solution and you 2 * 1000 mL of 0.02 M KMnO4 K 0.02 * 5 moles of H2O2
now know how much Ca2+ there is in 25.00 mL of solution.
2+ 0.02 * 5
●● You can now calculate the amount of Mg in 25.00 mL 1.0 mL of 0.02 M KMnO4 K moles of H2O2
of solution. 2 * 1000
Look at the second titration first: the MgCl2 reacts with the 48 * 0.02 * 5
48.0 mL of 0.02 M KMnO4 K moles of H2O2
EDTA left over from reaction with the Ca2+. Since the EDTA 2 * 1000
and MgCI2 solutions are the same concentration (0.0500 M) K 0.0024 moles of H2O2
then 1.0 mL of EDTA K 1.0 mL of MgCl2 solution: Therefore 25.00 mL of diluted solution contain 0.0024
Therefore 25 - 10 mL K 15.00 mL of EDTA solution moles of H2O2
must have reacted with the Ca2+ solution. Therefore 250.00 mL of diluted solution contain 0.024
1000 mL of 1.0 M EDTA solution K 1 mol of Ca2+ moles of H2O2
1000 mL of 0.0500 M EDTA solution K 0.0500 mol of Therefore 25.00 mL of 10-volume H2O2 contain 0.024
Ca2+ moles of H2O2
15 mL of 0.0500 M EDTA solution Therefore 1000 mL of 10-volume H2O2 contain 0.96 moles
15 * 0.0500 of H2O2
K mol of Ca2+ Concentration of 10-volume H2O2 is 0.96 * 34 g L-1 =
1000
32.64 g L-1 = 3.264 %
= 0.00075 mol of Ca2+
Therefore 50.00 mL of diluted urine solution contains
0.00075 mol of Ca2+ 28 Precipitation titrations
Therefore 25.00 mL of diluted urine solution contains 28.1 The equation:
0.00075/2 = 0.000375 mols of Ca2+ AgNO3(aq) + NaCl(aq) S AgCl(s) + NaNO3(aq)
Now look at the first titration:
Thus 1.0 mol of AgNO3 K 1.0 mol of NaCl
1000 mL of 1.0 M EDTA solution K 1 mol of metal ions
1000 mL of l.0 M AgNO3 K 58.5 g of NaCI
1000 mL of 0.0500 M EDTA solution
K 0.0500 mol of metal ions 1000 mL of 0.1 M AgNO3 K 0.1 * 58.5 g of NaCl
24.00 mL of 0.0500 M EDTA solution 0.1 * 58.5
1.0 mL of 0.1 M AgNO3 K g of NaCl
24 * 0.0500 1000
K = 0.0012 mol of metal ions
1000 10.3 * 0.1 * 58.5
10.3 mL of 0.1 M AgNO3 K g of NaCl
But from the second titration above, there are 0.000375 mol 1000
of Ca2+ ions in 25.00 mL of diluted urine solution. Therefore = 0.06025 g of NaCl
there must be
Therefore 25.00 mL of ‘potassium nitrate’ solution contains
0.0012 - 0.000375 = 0.000825 mol of Mg 2+ ions in 0.06025 g of NaCl
25.00 mL of diluted solution. Therefore 250.00 mL of ‘potassium nitrate’ solution con-
In 2000 mL of diluted urine solution there must be tains 0.6025 g of NaCl

% (w/w) NaCl in the ‘potassium nitrate’ is 0.8

0.6205 * 100
= 15.4646% = 15.5% 0.6
4.0124
Absorbance
0.4
29 Basic spectroscopy
29.1 Tip: Box 29.1 gives stepwise instructions for using a 0.2
spectrophotometer - check that you have covered all of the
major points. However, the exact details will vary from instru- 0
ment to instrument. 0 2 4 6 8 10
29.2 Tip: Substitute values into eqn [29.3] and solve for [C] (mar- Zinc concentration (mg mL-1)
gin examples are given on p. 219).
(a) Note that you must convert the path length from mm to cm
beforehand: 0.57 = 20 * 0.5 * [C] = 0.057 g L-1 = 31 X-ray fluorescence spectroscopy
57.0 mg mL-1 31.1 Tips: Write down the key points on how XRF works; ask a
(b) First calculate the concentration, using eqn [29.3]: colleague whether they understand the science of your expla-
0.31 = 20 * 1 * [C] = 0.0155 g L-1. Next, express the nation; prepare, practise and rehearse answers with your col-
concentration in the required units: 0.0155 * 109 (convert leagues.
to ng) , 106 (convert to mL) = 15.5 ng mL-1. Then, cal- 31.2 Interferences include:
culate the amount in 50 mL: 15.5 * 50 = 775 ng. (a) Spectral - occur from sources other than the analyte you
29.3 First determine the concentration of p-nitrophenol, in wish to measure.
mol L-1. Thus 8.8 mg mL-1 , 106 (convert to g mL-1) * 103 (b) Environmental - occur from lighter elements and can be
(convert to g L-1) , 291.27 (convert to mol L-1) = avoided by purging the instrument with an inert gas or
0.000 030 212 5 mol L-1. Next, substitute values into eqn creating a vacuum by the removal of air.
[29.3] and solve for e: 0.535 = e * 1 * 0.000 030 212 5, (c) Matrix - any element can absorb or scatter the fluores-
so e = 17707.902 36 = 17 700 L mol-1 cm-1. Note that you cence of the element, but can be corrected mathematically
should use the full numerical value until the final stage, to by alpha corrections.
avoid introducing ‘rounding’ errors. 31.3 Tips: When you are writing your protocol, consider wheth-
er someone else could pick it up and follow your instruc-
tions to achieve the same result. Use Box 31.2 as a guide
30 Atomic spectroscopy
but you will have to consider how you could ‘dry’ the soil
30.1 The calibration graph is shown below. The test solution
sample first.
has a K+ concentration of 0.32 mmol L-1. This is equiva-
lent to 0.32 , 1000 * 25 = 0.008 mmol in 25 mL (the
test sample volume). Next, divide by the weight of sam- 32 Chromatography
ple used in grams: 0.008 , 0.482 = 0.016 598 mmol 32.1 (a) Capacity factor for each compound using
(g sample) -1 = 16.6 mmol (g sample) -1. tR - tM
k′ =
0.8 tM
quinaldine = 4.9 and nicotine 5.2.
0.6
(b) Resolution using the following equation:
Absorbance
0.4 2[tRB - tRA]

Rs = is calculated as follows
WA + WB
0.2 2[6.2 - 5.9]
Rs =
0.16 + 0.18
0
0 0.1 0.2 0.3 0.4 0.5 0.60
+ Rs =
K concentration (mmol L-1) 0.34
Rs = 1.76
30.2 Calibration curve for a series of standards, assayed for K+
and based on the data in the table in Study exercise 30.1. (c) Average number of plates in the column using
See the graph below - note that this calibration line is ap- tR 2 tR 2
proximately linear up to ∙ 4.0 mg mL-1 but it becomes in- N = 16¢ ≤ or N = 5.54¢ ≤
wb w1/2
creasingly curved above this point. Water sample (a) con-
tains Zn at 1.70 mg mL-1; water sample (b) contains Zn at For quinaldine:
3.5 * 20 = 70.0 mg mL-1; water sample (c) contains Zn at 5.9 2 5.9 2
8.3 * 5 = 41.5 mg mL-1 (note that the graph curves at higher N = 16a b or N = 5.54a b
0.16 0.08
concentrations, making the estimate for water sample (c) less 2 2
reliable than the other two - a better approach might have N = 16(36.9) or N = 5.54(73.8)
been to dilute this sample further and reassay). N = 21786 or N = 30 173

For nicotine: Lysine : R@NH3+ ∆ R@NH2 + H +

2 2
6.2 6.2 low/neutral pH high pH
N = 16a b or N = 5.54a b
0.18 0.09 Note that the charge on a given amino acid side chain at a
N = 16(34.4)2 or N = 5.54(68.9)2 given pH will depend on the dissociation constant or pKa, (see
N = 18934 or N = 26300 p. 117) of the ionisable group.
33.3 In PAGE any diffusion of sample proteins that occurs reduc-
32.2 Test your knowledge of chromatographic theory
es resolution, so it is best to start with as narrow a sample
(a) Dead time (to) = amount of time the component spends
zone as possible. For most protein separations, the buffer pH
in mobile phase, essentially the dead volume of the col-
is chosen so that the proteins are negatively charged and will
umn. (Determined by injection of methane in GC.)
migrate from the cathodic end to the anodic end of the gel. In
(b) Retention time (tR) = time taken for a component
IEF, the separation is carried out on a pH gradient – because
to elute from the column. Measured from point of
proteins will all migrate to their pI values irrespective of the
injection.
tR - tM starting point, the position of sample application is not impor-
(c) Capacity factor = retention number k′ = tant. The only exception would be if any of the proteins to be
tM
(where tM is the dead time). separated were unstable at extreme pH values, in which case
(d) Separation factor a = tells us where peaks elute relative it would be best to avoid application at either end of the pH
k 2=
to each other: a = = gradient (e.g. pH 3.0 and pH 10.0). Once a given protein has
k1 reached the position corresponding to its pI, it will remain in
(e) Column efficiency N is a measure of peak narrowness: a narrow band because any diffusion away from the pI will
N = 5.54 (tr/w0.5)2 result in the protein acquiring a net charge and being focused
32.3 Calculate the resolution and efficiency of two compounds back to its pI.
(a) The resolution factor for the two peaks.
Rs = tr2 - tr1/0.5(w1 + w2) 34 Electroanalytical techniques
Compound X 20.63 - 18.40 34.1 These assumptions are given on p. 305. The electrical p otential
= measured at an ISE can be approximate to ion concentration
Compound Y 0.5(1.11 + 1.21)
assuming (i) the ion is in dilute solution (since the relationship
2.23
= = 1.92 = Rs is based on activity, eqn [34.3]; (ii)the ionic strength of the
1.16 calibration standards matches that of the test sample and
(b) The efficiency of each peak.
(iii) there is no substantial binding of the ion to sample com-
N = 16(tr/wb)2 ponents (e.g. to proteins or nucleic acids).
Using the above equation: 34.2 The two functional components of a biosensor are (i) an im-
Compound X = 16 (18.4/1.11)2 = 4397 mobilised enzyme (the biocatalyst) and (ii) a transducer, to
Compound Y = 16 (20.63/1.21)2 = 4651 convert the biochemical signal into the electrical signal that
32.4 Tip: Use eqn 32.4 to determine Rf values based on the meas- can be measured. A representative example (there are many
ured distances travelled by each component, relative to the others) would be the Biotrace luminescence-based system
solvent front. for monitoring microbial ATP in food, using luciferase (see:
Using eqn 32.4, the corresponding RF values are: http://www.biotrace.com).
pigment A = 63 , 114 = 0.553; 34.3 (a) 0.285 , 1000 * 4 (to calculate mmol in 4 mL) * 1000
pigment B = 76 , 114 = 0.667; (to convert from mmol to mmol) = 1.14 Mmol.
pigment C = 86 , 114 = 0.754. (b) 0.273 , 1000 * 20 (to calculate mmol in 20 mL)
32.5 (a) Flame ionisation detector. * 1000 (to convert from mmol to mmol) = 5.46 Mmol.
(b) Thermal conductivity detector. (c) 0.317 (estimated as the mid-point between the values at 14
(c) Electron capture detector. and 16°C) , 1000 * 10 (to calculate mmol in 10 mL)
(d) Diode array detector. * 1000 (to convert from mmol to mmol) = 3.17 Mmol.
32.6 Electrochemical detectors, then fluorescence detectors, then UV/ (d) 6.75 , 1000 * 250 (to calculate mg in 250 mL)
visible detectors. Many molecules, either intrinsically or after re- = 1.6875 = 1.69 mg.
action with a colour reagent, can be detected by UV/visible spec- (e) 4.73 (estimated as the mid-point between the values at 24
troscopy. However, fewer molecules show native fluorescence or and 26°C) , 1000 * 200 (to calculate mL in 200 mL)
the electrical properties necessary for electrochemical detection, = 0.946 mL.
making UV/visible detection the most versatile approach.
35 Using radio isotopes
35.1 Assume a 14C half-life of 5715 years (Table 35.2). Using eqn
33 Electrophoresis [35.1], ex = 0.5725, and so x = ln(0.5725) = - 0.5578. If
33.1 Tip: The dissociation of two important amino acids is shown -0.5578 = -0.693(t/5715), then t = ( -0.5578/-0.693) *
below: 5715 = 4600. The rat visited about 4600 years ago.
Glutamate : R@COOH ∆ R@COO- + H + 35.2 (a) 1200 Bq.
low pH neutral/high pH (b) 4.44 : 107 d.p.m.

(c) 1.20 MCi. 37.6 Ha: has 9NO2 in ortho position (sortho = 0.95) and 9OH
(d) 30.0 Bq g -1. in meta position (smeta = -0.12). Estimated chemical shift
(e) 28.0 pmol. d = 7.27 + 0.95 - 0.12 = 8.10 ppm. Hb: has 9NO2 in
The solution contains 250 * 10-6 * 5 mol = 1.25 *
35.3 (a) meta position (smeta = 0.26) and 9OH in para position
10-3 mol. The specific activity is therefore 55 * 106/ (spara = -0.45). Estimated chemical shift d = 7.27 +
1.25 * 10-3 Bq mol-1 = 4.4 : 1010 Bq mol-1. 0.26 - 0.45 = 7.08 ppm. Hc: has 9NO2 in para position
(b) 79.2 * 105/4.4 * 1010 = 1.8 * 10-4 mol per 2 h per (spara = 0.38) and 9OH in meta position (smeta = - 0.12).
107 cells is equivalent to 2.5 fmol s-1 cell-1. Estimated chemical shift d = 7.27 + 0.38 - 0.12 =
Therefore 25.00 mL of ‘potassium nitrate’ solution con- 7.53 ppm. Hd: has 9NO2 in meta position (smeta = 0.26) and
tains 0.06025 g of NaCl 9OH in ortho position (sortho = -0.56). Estimated chemical
Therefore 250.00 mL of ‘potassium nitrate’ solution con- shift d = 7.27 + 0.26 - 0.56 = 6.97 ppm.
tains 0.6025 g of NaCl % (w/w) NaCl in the ‘potassium 37.7 Ha has NH2 in ortho position (sortho = - 0.75), Br
nitrate’ is in the meta position (smeta = -0.13) and NO2 in the
0.6205 * 100 meta position (smeta = 0.17). Estimated chemical shift
= 15.4646% = 15.5% d = 7.27 + 0.17 - 0.75 - 0.13 = 6.56 ppm. Hb has NO2
4.0124
in ortho position (sortho = 0.95), NH2 in the meta po-
36 Infrared spectroscopy sition (smeta = -0.24) and Br in the para position
(a) Lots of sharp peaks–aromatic; sharp peak at (spara = -0.03). Estimated chemical shift d = 7.27 +
2200 cm-1 C ‚ N; benzonitrile. 0.95 - 0.24 - 0.03 = 7.95 ppm. Hc has NO2 in ortho po-
(b) Large peak at 1720 cm-1 C “ O; large peaks at 1250 cm-1 sition (sortho = 0.95), Br in the ortho position (sortho = 0.22)
and 1100 cm-1 C9O; ethyl ethanoate. and NH2 in the meta position (smeta = - 0.24). Estimat-
(c) Large broad peak at 3400 cm-1 O9H; 1-octanol. ed c hemical shift d = 7.27 + 0.95 - 0.22 - 0.24 =
(d) No functional group peaks, only … 3000 cm-1 and 8.22 ppm. (Very good agreement with spectrum.)
1460 cm-1/1375 cm-1 CH3/CH2 stretches and bends; 37.8 Ha split by Hb into a doublet (Jortho = 10 Hz) and each peak
heptane. split into another doublet (Jmeta = 2 Hz) by Hc.
Hb split into a triplet (Jortho = 10 Hz) by Ha and Hc and each
37 Nuclear magnetic resonance spectroscopy peak split into a doublet (Jmeta = 2 Hz) by Hd.
37.1 (i) Three types: CH3 (triplet), CH2 (sextet), CH2 (triplet). Hc split into a triplet (Jortho = 10 Hz) by Hb and Hd and each
(ii) Two types: 2 * CH3 (doublet), CH (septet). peak split into a doublet (Jmeta = 2 Hz) by Ha.
(iii) Three types: CH3 (doublet), CH (quintet), CH (doublet). Hd split by Hc into a doublet (Jortho = 10 Hz) and each peak
(iv) Molecule is symmetrical, therefore three types: 2 * CH3 split into another doublet (Jmeta = 2 Hz) by Hb.
(triplet), 2 * CH2 (quartet), 2 * CH2 (singlet).
(v) Molecule is unsymmetrical, therefore six types: CH3
(triplet), CH2 (quartet), CH2 (triplet), CH2 (triplet), CH2 38 Mass spectrometry
(quartet), CH3 (triplet). 38.1 CH3CH2COOCH2CH2CH3; a@cleavages:
37.2 (i) Two types: 3 * CH3 (quartet), C (singlet). (M - OCH2CH2CH3) = (M - 59) = m/z = 67,
(ii) Two types: 2 * CH3 (quartet), 2 * CH2 (triplet). (M - CH3CH2) = (M - 29) = m/z = 97;
(iii) Five types: CH3 (quartet), CH (doublet), 2 * C “ O (sin- b@cleavage: (M - CH2 “ CHCH3) = (M - 42) = m/z 84.
glet), 2 * CH2 (triplet), 2 * CH3 (quartet). CH3CH2CH2COOCH2CH3: a@cleavages:
(iv) Two types: CH3 (quartet), C (singlet). (M ¬ CH3CH2CH2) = (M - 43) = m/z = 83,
(v) Five types: para- CH (doublet), 2 * meta@CH (doublet), (M - OCH2CH3) = (M - 45) = m/z = 81;
2 * ortho@CH (doublet). b@cleavages: (M - CH2 “ CH2) = (M ¬ 28) = m/z = 98,
(vi) C (singlet), CH3 (quartet). (M - CH2 “ CH2) “ (M - 28) = m/z = 98.
37.3 The CH2 group has a phenyl substituent (s = 1.83) and 38.2 (a) Electron impact
a9C( “ O)OR (s = 1.46) so the estimated chemical shift is: (b) Chemical ionisation
d = 0.23 + 1.83 + 1.46 = 3.52 ppm. (The experimental (c) Atmospheric pressure chemical ionisation
value in CDC13 is 3.57 ppm: so good agreement.) (d) Elecrospray ionisation
37.4 The CH group has 2 * OEt groups (s = 1.14) and one
CH3 group (s = 2.50) so the estimated chemical shift is
d = 2.50 + 1.14 + 1.14 + 0 = 4.78 ppm. (The experi- 39 X-ray diffraction
mental value is 4.72.) 39.1 (a) Approximately 0.5 g
37.5 Ha: has Ph in gem position (sgem = 1.35) and CO2H (b) Yes, a pure powder
in cis position (scis = 1.35). The estimated chemi- (c) Yes, loose powders will give poor intensities
cal shift d = 5.25 + 1.35 + 1.35 = 7.95 ppm. Hb: 39.2 The father and son team of William H. Bragg and William L.
has Ph in cis position (scis = 0.37) and CO2H in gem Bragg.
position (sgem = 1.00). The estimated chemical shift 39.3 (a)
SiO2 glass is amorphous and hence does not have long-
d = 5.25 + 0.37 + 1.00 = 6.62 ppm. (The experimental range atomic order and hence produces only broad scat-
ones are 7.78 and 6.44 ppm respectively.) tering peaks.

(b) No, the silicon and oxygen atoms are arranged differently; 41.4 Tips: When writing a handwritten essay, the variant of the
the difference in their crystal tructure in reflected in the Vancouver method in which references are listed in the order
different X-ray diffraction patterns. cited might be the most convenient to use, because you could
Quartz Cristobalite write down all the references cited on a separate sheet as you
wrote out the essay. However, if you felt that it was important
for the reader to see the author(s) and publication dates of the
papers, then the Harvard method might be more appropriate –
however, you would need to wait until the essay was fully
written before organising the literature cited section in author
and date order. In a word processed review, you might prefer
the Vancouver method as this might allow the text to flow
more smoothly without being interrupted by lists of author
names and dates. On the other hand, the Harvard system is
40 Thermal analysis
easier to use as you write with a word processor, because you
40.1 The Mr of CuSO4.5H2O is 249.6
can simply ‘slot’ each new reference into the correct place
249.6 * 14.4 in the list. In an academic journal, it might be assumed that
First decomposition: = 35.9
100 the readership would want to know the authors and dates of
(corresponds to 2 H2O) the references cited, so the Harvard system would probably
249.6 * 14.8 be the most appropriate. In contrast, the journal Nature uses
Second decomposition: = 36.9 the Vancouver system in order to present important new sci-
100
(corresponds to 2H2O) entific advances to a wide audience. Nature readers want to be
able to assimilate each paper’s findings rapidly without get-
249.6 * 6.9
Third decomposition: = 17.9 ting distracted by citation details.
100
(corresponds to H2O)
Therefore final residue is 100 - 36.1 = 63.9% 42 Evaluating information
Therefore final residue is 249.6 * 63.9 = 159.49 = CuSO4 42.1 (a) P; (b) P; (c) P; (d) S; (e) P; (f) P; (g) P; (h) P; (i) S; (j) P;
(159.6) (where P = primary, S = secondary). Where journals con-
The thermal decomposition of CuSO4.5H2O involves relative- tain both original papers and reviews, we have classed them
ly easy loss of four molecules of water (ligand water) and then as primary.
one molecule of tightly bound water which is associated with 42.2 Tip: Discuss with a friend one of the topics. If one of you
the SO2-
4 anion. takes a ‘for’ stance and the other an ‘against’ stance, see how
many points each of you can make. Remember to try to back
41 Finding and citing published information up your view with scientific facts.
41.1 (a) The Dewey Decimal Classification system; the US Li- 42.3 Tip: Use Box 49.3 as a source to indicate ways in which
brary of Congress system; or other, as appropriate. graphs may be used to misrepresent information. You may
(b) Nature: Dewey: 505.0942; Library of Congress: Q1.N3; find this exercise easier to accomplish with printed graphs.
(or other, as appropriate). Note: the ‘Per’ or ‘per’ that may Those on television may not be shown for long enough to al-
precede these numbers refers to the fact that this is a peri- low you to carry out a detailed analysis. Nevertheless, they
odical, rather than a book. represent a rich source of material to criticise!
(c) Tip: Answer as appropriate – this will depend on the lay-
out of your particular library.
(d) (i) ‘A structure for deoxyribose nucleic acid’; (ii) ‘C60:
43.1 (a) Linus C. Pauling (1901–1994) is famous for winning
Buckminsterfullerene’.
two undivided Nobel Prizes; the first for Chemistry in
41.2 Tips: The main point here is that methods of writing down
1954 and the second for Peace in 1962. In winning the
references are diverse. You should also find that the details re-
Nobel Prize for Chemistry he was cited ‘for his research
corded are the same, although they may be in a different order
into the nature of the chemical bond and its application
or font style. You must pay attention to the precise instructions
to the elucidation of the structure of complex substanc-
given for your course, to ensure that you get this right when
es’. http://nobelprize.org/nobel_prizes/chemistry/laure-
writing up a project report or dissertation.
ates/1954/index.html (last accessed 01/03/16).
41.3 Tips: Three possible sites are:
(b) Dichloromethane (CH2Cl2) is also known as methyl-
(a) SI Units: http://physics.nist.gov/cuu/Units/units.html (last
ene chloride. http://www.environment-agency.gov.uk/
accessed 01/03/16).
business/topics/pollution/l92.aspx (last accessed 01/03/16).
(b) Diels–Alder reaction: http://www/organic-chemistry.org/
(c) The postal address of the London headquarters of the
namedreactions/diels-alder-reaction.shtm (last accessed
Royal Society of Chemistry is: Royal Society of Chem-
16/03/16).
istry, Burlington House, Piccadilly, London W1J 0BA.
(c) melting point determination: for example, http://www.wired
http://www.rsc.org/AboutUs/contacts/ (last accessed
chemist.com/chemistry/instructional/laboratory-tutorials/
01/03/16).
determination-of-meltingpoint (last accessed 16/03/16).

43.2 Tips: Note the number of hits found for each search engine. 45.2 See the figure below. Tips: When setting up the spreadsheet,
Make a note or printout of the top ten sites located by each create appropriate formulae in new columns for the trans-
search engine. Compare these site listings to reveal the quality formed data; graph using the ‘XY (Scatter)’ option in the Mi-
of data obtained. Repeat this exercise for meta-search engines. crosoft Excel Chart Wizard to make sure the data are correctly
Make lists of the component search engines used by the main spaced on the x-axis. Plot time on the x-axis and the trans-
meta-search engines to help you devise a strategy for the best formed cell count data on the y-axis. You will need to create a
coverage of the Internet. separate graph to examine the effects of each transformation,
43.3 Tips: Within the bookmark editor for your browser, cre- because the y-axis scales will be different. Select the Line
ate folders for each of your study courses and then create None option for the relevant (log transformed) series on the
sub-folders for each module that you are doing. Now use Data Series menu, then use the Add Trendline option (linear
some searches to put relevant bookmarks in each folder. Don’t model) within the Chart menu to add a linear regression line
forget to include a folder for transferable skills sites; there are (note: this can also be done using the Regression command
a lot of supportive sites available that are worth visiting. in the Data Analysis option on the Tools menu, but the pro-
cedures required are much more complex). Adjust the chart
44 Internet resources
output to suit, e.g. by changing gridlines, background and leg-
44.1 (a) Pr
ends, before copying and pasting to the MS Word document.
(b) 59
(c) lanthanides 1.2
(d) [Xe] 4f 36s2 1
44.2 (a) Royal Society of Chemistry
(b) 33.383 (this will vary annually) 0.8
(c) Douglas Stephen, University of Toronto, Canada 0.6

(d) Tutorial Reviews; Reviews 0.4
44.3 (a) A monosubstituted benzene ring
(b) Two similar intensity peaks, separated by a m/z of 2 0.2
(c) Two (79Br and 81Br with natural abundances of 50.69 and 0
0 100 200 300 400 500 600 700
49.31%, respectively)
times (s)
(d) The difference between the m/z of 77 m/z and 156/158 m/z
equates to a difference of 79 and 81. This coupled with the Decomposition of N2O5.
answer to (a) would allow you to conclude it is bromobenzene.
45.3 Tips: Take care when entering the date data in the spreadsheet.
45 Using spreadsheets
You may need to specify the format of the cells for the date
45.1 See the figure below. Tips: Depending on the precise method you
column (Format menu, Cells option). Highlight all parts of the
use, you may need to provide the Name, Values and Category La-
spreadsheet table including column headers, then use the Data
bels for the Chart Wizard or adjust those assumed. When creating
menu, AZT Sort option to rearrange data. You do not need to
the graph, select the ‘as new sheet’ option when prompted for a
fill in all the response boxes and can select ‘none’ if a box
Chart Location. For printing in black and white, select black and
contains criteria from a previous sort.
white hatching/shading options after click-selecting chart seg-
ments. Ensure the segments of the chart are labelled (i.e. not the
default legend option) using the Chart/Chart Options menu. 46 Word processors, databases and other packages
46.1 Tips:
potassium
2%
titanium (a) Ensure list components are in separate paragraphs; high-
magnesium 1%
light list; from the Table menu select AZT Sort function;
2% hydrogen
0%
ensure options are set to paragraphs, text, ascending.
sodium
calcium 3% (b) From the Edit menu, select Replace option, then fill in the
4% relevant boxes.
iron
6% (c) From Edit menu, select Replace option, then fill in the
relevant boxes; select More▼ then Format, Font, Italic.
(d) From the View menu, select Header and Footer and use
oxygen the small on-screen menu to add text and numbering as
aluminium
8% 46% desired. Formatting commands (e.g. centre) will work in
the header and footer boxes. As an alternative for adding
page numbers, this can be done from the Insert menu,
Page Numbers . . . option.
(e) Margins can be adjusted from File menu, Page Setup,
Margins option.
silicon (f) Select paragraphed text, then use the Format menu, Bul-
28% lets and Numbering option, using Customise if the option
Relative percentage composition of elements in the earth’s crust. you desire is not displayed within the panel.

(g) Type ‘alternative’ within a Word page. Ensure cursor is (a) The main physical forms encountered are likely to be solid
within word (or highlight word); from Tools menu select or liquids (though gaseous samples might also need to be
Language, Thesaurus options. Perhaps ‘option’ might be considered). In general most instrumental analytical tech-
a useful synonym? niques require samples to be in liquid (aqueous or organic
(h) From Tools menu, select Spelling and Grammar to start a solvent) form; on that basis some sample preparation is
spell check. Take care if the checker queries UK spellings normally required to convert a sample into a suitable form.
(it may be set to accept US spellings). Note: this is not a (b) Determination of the linear calibration range for a particu-
substitute for reading your document – the spell checker lar instrumental analytical technique is necessary to obtain
will miss typing errors like ‘form’ instead of ‘from’. accurate and precise data. The linearity of your calibration
(i) For whole document, triple click in left margin to select graph is very strongly influenced by your manipulative
all, then use Tools menu, Word Count option. For selected and manual dexterity skills in preparing standard solutions.
text, highlight (select) the part you wish to count, then (c) Most techniques have some interferences issues that you
carry out same instructions. need to be aware of; sometimes as part of avoidance is
(j) While in first document, use File menu, Open option to better than cure or the required for additional sample
open another. You can switch between them using the Win- preparation regimes to minimise potential interferents.
dow menu, where you will see all open documents listed. (d) A range of approaches are available and are discussed in
Note: remember to save changes to all documents when Chapters 30–31 and 32 specifically.
closing down your editing session.
46.2 Tips: Although there are several ways to modify tables, you 48 Calibration and quantitative analysis
may find it useful to try the following commands (think care- 48.1 Determine unknowns from a calibration curve produced
fully about their order). in Excel.
●● Table, Insert Table Determination of chlorophenol in an extract
12000
●● Table, Merge Cells
●● Table, Distribute Columns Evenly
10000
●● Tables and borders, modify borders commands (various)
Consult Box 50.2 or use the Help feature if you run into 8000
Peak area
problems.
6000
46.3 Tips: You might start your search on the networked ‘desktop’
and icons visible to you when you log on. Use ‘help’ menus 4000
and ‘wizards’ to investigate how to use the programs, or con-
sult any manuals available in the computer suite. 2000
47 Fundamental principles of quantitative chemical 0

0 2 4 6 8 10 12 14 16
analysis Concentration (mg/L)
47.1 (a) Accuracy – the closeness of a measurement to the true (a) 1.18 mg L–1; (b) 3.16 mg L–1; (c) 5.19 mg L–1.
value.
(b) Precision – the closeness of measurements to each other. 48.2 Tip: Check that you understand the general requirements for
(c) Quality control – measures in place to ensure that the good graph drawing, detailed in Chapter 49.
results generated meet your laboratory’s standards. (a) The title of the figure (‘calibration graph’) does not give
(d) Quality assurance – measures in place to monitor and doc- sufficient information to enable the reader to understand
ument the performance of a test procedure. the content.
(e) Replicate – a repeat measurement. (b) No details are given as to the nature of each set of data
(f) Detection limit – the minimum concentration of an analyte values – are these curves for two separate substances?
that can be detected at a particular confidence level. (c) The x-axis has a poor choice of units – it would be better to
47.2 As a discussion point with colleagues you may find that use mg, then all of the values would be 1000 times smaller.
UKAS accreditation could either be a positive attribute or a (d) The x-axis runs beyond the highest value (8 mg), wasting
negative one. Some points to consider are: around 20% of the horizontal space.
●● Would your work always want to adhere to standard oper- (e) The scale marks (tic marks) on the x-axis are missing.
ating procedures? (f) The x-axis has no zero shown.
●● Would you always use certified reference materials in your (g) The y-axis has too many numbers, and too many minor
work? scale marks – it is probably better to have a tic mark every
●● How would you establish and maintain quality assurance 0.1 absorbance unit. (Note that absorbance (A) is a dimen-
procedures? sionless term, so no units are required – this is not an error.)
●● Is your recording of results in your lab note book always (h) The data values represented by the square symbols show
up-to-date? It would need to be for UKAS accreditation. a reasonable fit to the lower linear trend line, but the data
47.3 The responses to these specific questions require some spe- values represented by the round symbols do not – the lat-
cific instrumental analytical technique knowledge. You might ter set of data shows a clear curvilinear relationship and
need to refer to Chapters 29–35 inclusive for some guidance. therefore a linear trend line is not appropriate.

48.3 Tip: Based on your responses to 48.1 and 48.2 you should 30
be able to create a properly labelled calibration graph using
linear regression. 25
49 Using graphs 20
49.1 (a) Three-dimensional graph
Frequency
15
(b) Pie chart
(c) Scatter diagram with an added trend line
10
(d) Plotted curve
49.2 other 5
fibre 2 g protein
4g 6g 0
7–7.9 8–8.9 9–9.9 10–10.9 11–11.9 12–12.9 13–13.9 14–14.9 15–15.9 16–16.9
fat Fructose content (g L-1)
5g Frequency distribution of data of table in study exercise 49.3.

49.4 Tip: The Sunday newspaper supplements often analyse news
stories with the aid of graphs, and might be a good source for
material, as their articles are often slanted towards a particular
viewpoint, and you may find that the graphical presentation
carbohydrate has been chosen to support this.
83 g
50 Presenting data in tables
50.1 Concentrations of drug and metabolites found in a blood sam-
ple (mg L-1); alcohol expressed in mg per 100 mL-1.
Typical composition of a breakfast cereal
Methadone 0.23
49.3 Tip: First decide on the class boundaries and number of Diazepam 0.2
classes – in this case, the values range from 8.1 to 15.1. The Desmethyl diazepam 0.075
subdivisions shown in the table below provide eight classes, Temazepam 0.1
which gives a frequency distribution with a reasonable num- Nicotine NQ
ber of classes to aggregate the data sufficiently, but also pro- Caffeine NQ
vide enough detail to indicate the overall shape of the distri-
Alcohol 86
bution. Note that Excel also has a useful histogram function,
but you will need to learn how to set the class boundaries NQ = positive result, but not qualified
(termed ‘bin values’) to make effective use of this function
50.2 Tip: Box 50.1 gives the key aspects. The following table gives
(see Box 49.2).
one possible layout.
A histogram for these data is shown in the figure over-
leaf. Your brief description should include something about The principal components of a typical table
the location, dispersion and shape of the distribution (see
Chapter 52), for example: ‘The modal class of the distribution Component Comment
is 12.0912.9 g L-1, with a range of 7.0 g L-1(15.598.5 g L-1). Title Concise, self-contained description of
The data show a slight negative skew, with 28 values in the three table contents, with numbering where
classes above the mode and 46 in the four classes below it’. appropriate
Data of table in study exercise 49.3 Column/row Identifies the content of each column
presented as a frequency distribution headings and row, showing units of measurement,
where appropriate
Class Frequency Data values Quoted to an appropriate number of
8.0–8.9 2 significant figures/decimal places
9.0–9.9 5 Footnotes All abbreviations and other individual
10.0–10.9 17 details must be explained
11.0–11.9 22 Rulings Used to emphasise any groupings within
the table, and to separate individual items
12.0–12.9 26
13.0–13.9 17
14.0–14.9 10 51 Hints for solving numerical problems
51.1 (a) [C] = A/el
15.0–15.9 1
(b) e = A/[C]l

51.2 (a) Subtract ax from both sides and then swap sides, so: Relative frequency Hyphal length m(m)
b ∙ y ∙ ax (% total)
(b) 1. Subtract b from both sides and then swap sides, so:
ax = y - b Treatment A Treatment B
2. Divide both sides by a, so: x ∙ (y ∙ b)/a 5 5.2 0.7
(c) Raise each side to the power 13, then swap sides, so:
y ∙ x1/3 6 8.7 1.7
(d) 1. Take logs of both sides, so: log x = log(3y) 7 14.3 3.1
2. Noting log(3y) = y log 3, substitute on right-hand side,
so log x = y log 3 8 19.0 4.9
3. Divide both sides by log 3, then swap sides, so: 9 18.3 7.3
y ∙ log x/log 3
(e) 1. Divide both sides by 1 - y, so: z p + 3 = x/(1 - y) 10 12.7 9.0
2. Subtract 3 from both sides, so: z p = x/(1 - y) - 3 11 7.1 10.8
3. Raise both sides to the power 1/p, so:
12 4.4 11.5
z ∙ ((x ∙ (1 ∙ y)) ∙ 3)1/p
(f) 1. Multiply both sides by pq, so: zpq = (y - z)1/n 13 2.4 10.4
2. Raise each side to the power n, so: zpqn = (y - z)
14 1.2 9.0
3. Take logs of both sides, so: nlog(zpq) = log(y - z)
4. Hence n ∙ (log(zpq)) ∙ (log(y ∙ z)) 15 0.4 7.6
51.3 (a) 215
16 0.4 6.3
(b) 107 000
(c) 0.04 17 0.0 4.9
(d) 99.82 18 0.0 4.2
(e) 99.90
(f) 100.00 19 0.0 2.8
(g) 6260 20 0.0 1.7
(h) 130 000
(i) 2 3 or 4, depending on circumstances (see p. 491) 21 0.0 1.4
(j) 5 22 0.0 0.7
(k) 3
(l) 6 23 0.0 0.7
(m) 2 24 0.0 0.3
(n) 5
51.4 (a) 17.14% 25 0.0 0.3
(b) 7.55%
(c) 14.01
(d) 100 * (75.02 - 55.23) , 55.23 = 35.831 975 =
35.83%
52 Manipulating and transforming raw data

52.1 You first need to work out relative frequencies (see table be-
low), then graph the data appropriately (see figure below).
Relative frequencies of hyphal lengths in treatments A and B
Relative frequency Hyphal length m(m)

(% total)
Treatment A Treatment B
0 0.0 0.0
Frequency polygons for data in above table.
1 0.4 0.0
2 0.8 0.0 52.2 Mean: 7.60; standard deviation: 0.886.
3 1.6 0.3 52.3 Tip: The frequency distribution of the untransformed data
indicates that they are positively skewed (see figure). The
4 3.2 0.3 ladder of transformations shown in the figure suggests initial

corrections in increasing strength being square root x and 53.2 Tip: Substitute values into eqn [53.3] to calculate the SE of
log(x). Figure (b) indicates that the square root transformation each sample.
is approximately symmetrical, whereas figure (c) indicates For sample A: 12.7 , 3.464 101 615 = 3.666 174 209 =
that the log transformation results in a slight negative skew, in 3.67.
effect ’over-transforming’ the data. For sample B: 14.4 , 4.472 135 955 = 3.219 937 888 =
3.22.
Thus, in absolute terms, sample B has the lower standard er-
ror (note that the higher standard deviation of sample B is
more than offset by the increased number of data values). To
calculate the proportional standard error, divide the SE by the
mean, as follows:
For sample A: 3.666 174 209 , 16.2 = 0.226 307 05 =
0.226.
For sample B: 3.219 937 888 , 13.2 = 0.2439 346 88 =
0.244.
Thus, in proportion to the mean, sample A has the lower
standard error.
53.3 Vitamin concentration data (recalculated)
Mean vitamin
Replicate concentration Sample Mean size :
sample (mg) number sample size
A 3.0 24 72.0
B 2.5 37 92.5
C 2.0 6 12.0
Total 67 176.5
True mean = 176.5/67 = 2.63 mg
54 Choosing and using statistical tests

54.1 Tip: First calculate the standard error using eqn [53.3], as
6.8 , 4 = 1.7. Then, substitute values into eqn [54.2] to cal-
culate the 95% confidence limits (note that you also need the
value for t0.05[15] = 2.131). The upper limit is calculated as:
24.7 + (2.131 * 1.7) = 28.3227 = 28.3. The lower limit is
calculated as 24.7 - (2.131 * 1.7) = 21.0773 = 21.1.
54.2 Tip: The calculated t ∙ 2.136 with 12 degrees of freedom;
the critical value at P = 5% is 2.18 (Table 64.2); hence you
would accept the null hypothesis that the samples came from
the same population and hence conclude that the different an-
alytical methods had no significant effect.
54.3 Tip: Amino acid uptake will be the dependent (y) variable and
time the independent (x) variable. From the Excel output, the
coefficients are: intercept = 1.17 (to 3 significant figures)
and slope (labelled as ‘X variable 1’) = 2.95 (to 3 signifi-
cant figures), so the linear relationship has the following form:
Effect of different transformations on distribution of data from Treat- y = 1.17 + 2.95x. The strength of the relationship is meas-
ment B in hyphal growth experiment. (a) Untransformed data; (b) ured by the coefficient of determination, termed ‘R squared’
square root transformation of (x); (c) log transformation of (x) in the Excel output. In this case, a value of 0.971 (to 3 signifi-
cant figures) shows that there is a very strong fit of the y data
53 Descriptive statistics to the trend line.
53.1 (a) Range ∙ 9
(b) Variance ∙ 4.46
(c) Standard deviation ∙ 2.11
(d) Coefficient of variation ∙ 43.1% (CoV = 100 *
2.113/4.9)
(e) Standard error ∙ 0.273

55 Drawing chemical structures value of 243 kJ mol-1. As the two chlorine atoms are brought
55.1 (a) CH3CH2CH2CH2OH CH2Br closer than the equilibrium distance, the energy increases very
(b) (c) CH3CHCH3
rapidly due to the electrostatic repulsion between the two pos-
NH2 itively charged nuclei.
58 The importance of transferable skills

O 58.1 Tips: Where you feel confident about skills you have
O
learned at school, take care to consider whether they might
S Cl
(d) CH3C OCH2CH3 (e) need to be upgraded for university/college use. When think-
ing about opportunities for developing skills, remember
O
CH3 that these may occur outside university, perhaps in work or
social contexts.
55.2 O O O 58.2 Tip: Possible keywords/phrases for a Web search include:
(a) (b) Cl (c) (d) chemistry skills, C&IT skills, numeracy skills, study skills,
OH O time management.
58.3 Tip: You could create a grid to record your thoughts, with col-
umn headings ‘University’; ‘Work’ and ‘Social’ and rows for
(e)
‘Short term’, ‘Medium term’ and ‘Long term’. Short term and
medium term could refer to your time at university, while long
55.3 (a)
Three bonding pairs, no lone pars, therefore trigonal
term could refer to after graduation.
planar.
(b) Two bonding pairs, two lone pairs, therefore tetrahedral 59 Managing your time
based. 59.1 Tips: Model your spreadsheet on Fig. 59.1. It isn’t too difficult
(c) Two bonding pairs therefore linear. to fill in details at the end of each day either directly into the
(d) Six bonding pairs, therefore octahedral. spreadsheet or on a printed version. You will need to collect
(e) Five bonding pairs and one lone pair, therefore octahedral information for at least a week and possibly longer before you
based. see patterns emerging.
59.2 Tips: It is important to include both social and study-related
56 Chemometrics activities on your lists. Short-term lists deal with issues of
56.1 Tip: Use the information provided in Chapter 41 to identi- the day or week; medium-term for the term or semester and
fy a scientific paper that has used either a factorial or central longer-term for one year onwards. Relate each of these lists to
composite design. The boundaries of their chosen variables appropriate goals you may have over these timescales.
may have been determined by some preliminary experiments 59.3 Tips: This task could be related to (a) your self-analysis of
or because of practical limitations, e.g. it was not possible to transferable skills (Chapter 58), or (b) to some large task such
operate under the conditions. as carrying out a final-year project. In the latter case, for ex-
56.2 Tip: Use the information provided in Chapter 41 to identify a ample, there are small jobs that can be done as you proceed
scientific paper that has used a principal component analysis with the larger task, such as writing parts of the Introduction
(PCA). The visualisation of large data sets to identify trends or compiling a list of references, and these will reduce time
can be effectively done using PCA. pressure towards the end of the task.
56.3 Tip: Discuss with other students or your project supervisor
how you might use optimisation approaches to assist with 60 Working with others
identify best operating conditions, e.g. designing a new or- 60.1 Tips: Think about your contributions to past group activities
ganic synthesis and wanting to determine the optimum condi- while doing this exercise, and, if necessary, get feedback from
tions for yield or to maximise the signal output from an instru- friends and colleagues. You may feel that you fit into more
mental analytical technique. Also how might you use PCA to than one ‘natural’ role: this is quite often the case.
visualise a large data set. 60.2 Tip: This exercise might best be associated with a specific
teamwork activity, but this need not be restricted to study –
57 Computational chemistry group contributions with clubs, societies or in employment are
57.1 The two methods both give equilibrium bond distances close equally valid. Strategies for improvement will depend on your
to the experimental value. However, the molecular mechan- identified weaknesses. For example, if you felt too shy to con-
ics calculations fail at distances beyond ∙ 2.1 A ∘ , because they tribute effectively, you might want to practise public speaking
predict that as the Cl2 molecule is pulled apart, the energy (see Chapter 67) or attend an assertiveness workshop.
again increases very rapidly (like a very strong spring being 60.3 Tip: Columns 4 and 5 of Table 60.1 are relevant, particularly
stretched). In fact, as the atoms are pulled apart, the Cl ¬ Cl if you are able to identify a ‘natural’ role or roles for yourself
bond starts to break, so the energy tails off to the value for two in group situations (as in Exercise 60.1). Can you relate your
isolated chlorine atoms. The DFT method models this reason- strengths and weaknesses to specific events that have occurred
ably accurately; the curve levels off to give a bond dissoci- during teamwork, and might you handle the situation differ-
ation energy of 200 kJ mol-1, compared to the experimental ently in future?

61 Taking notes from lectures and texts 64.2 Tip: When revising, try to find a topic you find hard to under-
61.1 Tips: If you are used to using a particular note-taking method stand and see whether your colleague has approached it in the
that seems to work well, you may feel reluctant to experiment. same way as you. If not, you may be able to learn from this.
Try out the new method by taking notes at a seminar, tutorial 64.3 Tip: Discuss your tactics with a colleague sitting the same
or meeting where it will not matter so much if the technique exam. What topics do you agree are likely to come up?
does not work well at the first attempt.
61.2 Tip: Make a list of any missing notes or handouts and ask a 65 Preparing your curriculum vitae
colleague if you can work from their notes to fill in these gaps. 65.1 Tip: This exercise might be attempted with a friend you can
61.3 Tip: To compare methods, test your recall some time after- trust to give you a frank opinion. Remember that your per-
wards. Take a blank sheet of paper and see what you can write sonal qualities can be developed in both curricular and extra-
about each topic. curricular activities.
65.2 Tip: Working with a friend or group from your class, compare
62 Learning effectively notes on how you have organised your CVs and then modify
62.1 Tip: When considering how to adapt to your apparent learning your own CV, taking on board good ideas from your peers.
preference, bear in mind that diagnostic tools are not perfect Create a physical file for your revised CV and add handwrit-
and, especially if your preferences are mixed or ’multimodal’, ten notes, updating it as and when required.
there may be a range of techniques that could suit you. Don’t 65.3 Tip: Use the ‘Prospects’ website noted in the sources for fur-
be afraid to experiment, but follow your feelings about what ther study to gain ideas before your appointment with the ca-
’feels right’ and do this, rather than what any text or website reers service. The library or careers resource centre may have
advises. useful information, often accessible via the Web.
62.2 Tip: If you are worried that trying out a new method of
note-taking (say mind-mapping) might result in notes that are 66 Organising a poster display
meaningless after the event, why not try out the method in a 66.1 Tip: Use Figure 66.1 and the subheadings of Chapter 66
’low stakes’ situation – say a general interest lecture, a meet- as a starting point for assessing the pros and cons of your
ing or while watching a TV programme. designs.
62.3 Tip: Use the cue words in the question instructions to under- 66.2 Ten-point checklist – what makes a good poster presentation?
stand better what your lecturers are expecting and the exam- □ Title – is this concise, specific and interesting?
ples in Table 5.1 to reflect on your own approach to the ques- □ Author(s) – does the poster show details of the names and
tion. addresses of all contributors?
□ Structure – is the overall structure clear to the observer,
63 Revision Strategies with good use of subheadings?
63.1 Tips: When constructing your revision timetable, remember □ Flow – is the layout of the poster clear, in terms of how the
the following important points: audience is expected to ‘read’ each section?
●● Break large topics into ‘bite-sized’ units. □ Content – does the presentation have a clear introduction,
●● Mix up hard or less interesting topics with those you enjoy main message and conclusion?
revising (don’t ignore them). □ Text – does the poster contain an appropriate amount of
●● Keep an appropriate balance between work and relaxation. written material in a ‘readable’ font?
●● Offer yourself rewards if work targets are completed. □ Graphics – does the poster make good use of diagrams,
63.2 Tips: You may wish to keep one of the past papers (preferably figures and images?
a recent one) aside to use in a mock exam. Remember that □ Size and scale – does the poster make effective use of the
learning objectives can be just as important as exam papers in space available?
helping you to revise. □ Colour – does the presentation use colour effectively?
63.3 Tips: Read through the list of active revision tips on p. 558 □ Conclusion – is the ‘take-home message’ clear to the
and decide which might work best for you. Carry out a tri- observer?
al of the method with one of your topics where the material 66.3 Tip: You might use the ten-point checklist given above, or
has seemed difficult to learn. Experiment until you arrive at a your own version from exercise 66.2 to assess selected posters
solution that works, remembering (a) that different techniques by giving a mark out of 10 for each item in the list. In a group
might work best in different subjects and (b) a little variety exercise, you can see how your ‘scores’ compare with those of
might make the revision process more interesting. other students, and discuss items where scores are different.
64 Assignments and exams 67 Giving a spoken presentation

64.1 Tip: Some questions to ask yourself about past exam perfor- 67.1 Ten-point checklist – what makes a good spoken presenta-
mances… Did you prepare well enough? Did you run out of tion?
time during the exam? Did you spend too long on a particular □ Introduction – does the speaker explain the structure and
question or section, and have to rush another? Were your an- rationale of the topic at the outset?
swers direct and at the appropriate depth? Did you misinter- □ Audience – is the material presented at an appropriate
pret any questions, or miss out part of any answers? Were your level?
writing skills, including planning, up to the standard required? □ Content – does the presentation cover the topic effectively?

Speed and structure – are the main elements of the pres-

□ 69.2 Tips: Compare your ideas with those of a partner, or ask tutors
entation presented clearly, and at an appropriate pace? or lecturers what they think of your efforts. A useful adjunct
□ Voice – is the presentation delivered in an engaging, active to this exercise is to write the first sentence of your answer to
vocal style? avoid writer’s block (p. 598).
□ Audio-visual aids – does the speaker make good use of 69.3 Tips: Note that each of the stages (b)–(d) requires a separate
visual images (e.g. graphs, photographs)? ‘review’ – it is difficult, if not impossible to make a good
□ Image – does the presenter make good use of body job of all aspects in a single ‘pass’ through the text. Ideally,
language? you would reprint the essay between reviews, having made
□ Notes – does the speaker use notes to structure the talk, corrections.
rather than reading from a written text?
□ Conclusion – is a clear ‘take-home message’ delivered in 70 Reporting practical and project work
the final stages of the presentation? 70.1 Tips: To assimilate the required style for Experimental, you
□ Questions – does the presenter deal with questions may find it appropriate to read some research papers from
effectively? the journal section of your library. Note particularly that
67.2 Tips: You could try marking speakers out of 10 for each of Experimental is a description of ‘what was done’, not a set of
the points listed in exercise 67.1 (either using the checklist instructions.
provided above or your own version), or you could simply 70.2 Tips: Again, you may find it appropriate to read some research
prepare a list of the good and bad points of the presentations papers from the journal section of your library to assimilate
you have evaluated. the normal style for describing research results.
67.3 Tips: When evaluating your own performance: (i)remember 70.3 Tips: Abstracts are difficult to write: try to include a synop-
that most of us are overcritical when viewing ourselves on sis of all the main sections of the paper, including its aims,
video, tending to focus on relatively minor points, e.g. facial methods, key results and conclusions. Ignore the word length
expression, rather than broader aspects, e.g. clarity of deliv- restriction until you have noted the main points, then try to
ery and content; (ii) when giving a talk to a group of fellow reduce (or, less likely, expand) your effort to match the re-
students you might ask them to use all or part of the checklist quirement.
from exercise 67.1 as a way of providing feedback.
68 General aspects of scientific writing 71 Writing literature surveys and reviews

68.1 Tips: When brainstorming, think of all aspects of the topic 71.1 Five differences between reviews and papers are:
that could be relevant (even if you could not give precise de- (1) Reviews deal with material in general terms taking an
tails at this stage). When you meet together, discuss which of overview, whereas papers tend to look at a focused, single
your combined ideas would be most important in crafting an aspect of a topic.
answer and what order you might place them in. (2) The subdivisions are generally not the same – papers
68.2 Tip: Try to apply what you have learned in the next assign- usually follow the IERaD format (p. 602), while reviews
ment you write – ask a tutor for specific feedback if you are are structured according to the major components of the
unsure about whether your technique has improved. topic.
68.3 Tips: Use the spelling list to help you when writing and make a (3) There is little or no reference to original data in reviews.
conscious effort to memorise the spelling each time you use it. (4) There is little detail of methods in reviews.
The act of writing the vocabulary list will assist you to memo- (5) They tend to use different reference citation systems, with
rise new words and their meanings. If you have trouble learn- Harvard (p. 382) most commonly used in papers and Van-
ing definitions, e.g. terms in chemistry, you could extend these couver (p. 382) used in reviews.
ideas to create a personal glossary of terms and acronyms. 71.2 Tip: It should be possible to do the second part of study ex-
ercise 71.2 online at your library – you may need to enlist the
69 Writing essays help of library staff if unsure of how to do this.
69.1 Tip: Can you determine what lies behind each question? Some 71.3 Tip: Obviously, the Abstract and Conclusions sections will
question titles are quite direct, but others ask for information provide information about the key points addressed. It might
in a more subtle way. Use your imagination to think beyond be helpful to use a highlighter on a photocopied version of the
the immediate question, linking to learning objectives or to review to note important statements within the main body of
other relevant material covered in lectures. the review.

Index
A attributes, measurement of, 36 capillary gel electrophoresis (CGE), 299–300

abbreviated formulae for chemical structures, 502 authorship, evaluation sources of, 384–5 capillary isoelectric focusing (CIEF), 300
ab initio method, 514 autoradiography, 313–14 capillary tubes, 124–5
absorbance, 217, 220 azeotropes, 145 capillary zone electrophoresis (CZE), 298
abstracts, 377 carbon skeleton, abbreviations, 502
accelerated solvent extraction (ASE), 276 B cations
accuracy, 38 back-loading method, 364 flame tests for, 182
acetates, printing on, 5 back titrations, 204–5 testing in quantitative analysis, 179–80
acid–base extraction chemistry, 139 balances, 65 Celsius scale, 37
acid–base reactions, 107–8 balances and weighing, 78–80 Central Limit Theorem, 491
in volumetric analysis, 190 analytical balances, 79 centrifugation
acid–base titrations, 199–202 general purpose balances, 78–9 low-speed, using, 181
indicators for, 199 bar charts, 449 of solutions, 180–1
neutralisation curves in, 199–200 base, 113 Cerencov radiation, 313
standardisation of, 200–2 beakers, 64, 67, 92 certified reference materials (CRMs)
using KPH, 306 becquerel, 311 and sample size, 229–30
acid digestion of solids decomposition, 238–40 Beer–Lambert law, 217 charcoal, removal from solution, 87
active revision, 554 Belbin, R.M., 535 checklists, 531
activity of solutes, 104–5 beta decay, 309 chemical interferences in AAS, 232
adsorption chromatography, 250 bias in measurement, 38 chemicals
agarose, 289–90 bibliography, 379–80 light sensitive, storing, 67
air displacement pipettors, 64 bimodal frequency distributions, 483 using, 70
algebraic rules, application of, 465 binomial distributions, 488–9 safety, 70
alkali, 113 biosensor, 307 selection, 70
alpha decay, 309 blogs, 399 chemical shift
ampholyte, 113 blotting, 295 correlation tables, 337–9
Amsterdam Density Functional (ADF) bond-line formulae for chemical in 1H@NMR spectra, 335–7
package, 515 structures, 502 in NMR spectroscopy, 333
analysis of variance (ANOVA), 494 bookmarks, 392 chemical synthesis, 562
Analytical Abstracts, 376 bottles and vials, 67 chemistry, internet resources for, 399–420
analytical writing, 591 Bragg’s Law, 365–6 databases, 401–19
anions, testing for, 179–80 brainstorming, 535, 537 principal resources, 400
aqueous solutions Bravais lattices, 366–7 chemistry-related practical skills, 524
IR spectra of, 322 Bremsstrahlung, 245 CHEMnetBASE, 408
quantitative analysis of, 74 bromine atoms in a molecule, 355 chemometrics, 507–12
argentimetric titrations, 211 Buchner funnels, 84–5 experimental design, 508–10
assessed coursework, 556–7 Buckingham, J., 128 factorial design, 509
asymmetry factor in chromatography, 253 buffer solutions, 116 methodology, 508–10
asymmetry in frequency distributions, 482 bumping, 87 principal component analysis, 511–12
atmospheric pressure chemical ionisation Bunsen burners, 87–8 response surface design, 509–11
(APCI), 358 burettes, 64 sequential design, 508
atomic absorption spectrophotometry (AAS), filling in volumetric analysis, 193 simultaneous design, 508
226–33 burners, 87–8 ChemSpider, 406–7
atomisation cell, 227–228, 230 chiral capillary electrophoresis (CCE), 298–299
background correction, 232 C chlorine atoms in a molecule, 355
interferences, 232 calculators, 4 chromatography
principle, 231 calibration in quantitative analysis chromatogram, 252–3
radiation sources, 226 curves detectors, 254–5
sample introduction, 230–1 stages involved in preparing and using, 442 ionisation sources, 357
wavelength selection, 231 standards, 443 mass spectrometry in, 357
atomic orbitals, drawing, 503 types of, 441 resolution, 253–4
atomic spectroscopy, 222–4, 226–40 plotting, 223 separation methods, 250–2
atomic absorption spectrophotometry calomel electrode, 303–304 types of, 255–6
(AAS), 226–33 Cambridge Structural Database (CDS) citations, 378
atomic emission spectrophotometry (AES), 233 System, 366 clamp holders, 82
inductively coupled plasma, 234–5 capacity and effects of pH, 117 clamps, 82–3
inductively coupled plasma mass spectrometry, preparing, 118–19 Clark-type oxygen electrodes, 306
13
235–8 selecting, 117–18 C@NMR spectra, 343–6
attenuated total reflectance (ATR) sample capacity factor in chromatography, 252 coefficient of variation, 481–2
holders, 323, 325 capillary electrophoresis, 297–300 collaboration for learning, 537
Index 627

Index
colour for poster display, 576–7 Debye-Scherrer X-ray diffraction electrolyte, 102
column efficiency in chromatography, 253 camera, 366 electrolytic cell, 305
combinatorial chemistry decolorisation in recrystallisation, 134 electrolytic dissociation, 102
parallel synthetic method, 174–5 DeepView package, 515 electronic configuration, 503–4
solid phase synthesis, 173–4 degrees of freedom, 488 Electronic Laboratory Notebooks (ELNs), 53
communal databases, 54 density functional theory, 515 electron multiplier tube (EMT), 359
comparative writing, 591 derived data, working with, 38 electro-osmotic flow, 289
complexation reactions, 108 design for poster display, 575–6 electrophoresis
complexometric titrations, 203–7 dessicators, 96–7 definition, 287
back titration, 204–5 detectors separation of proteins, 287–8
calculations, 206–7 in inductively coupled plasma supporting medium, 288–9
direct, 204 spectroscopy, 235 electrospray (ES) ionisation, 358
ligand types, 203 for mass spectrometry, 359 elemental microanalysis, 187–8
procedures, 205 deuterated solvents, 334 employability, 527
using EDTA, 203–4 diagrams in essay writing, 595 empty cells in spreadsheets, 423
computational chemistry differential scanning colorimetry, 370 end-point determination in precipitation
computational laboratory, 516–18 differential thermal analysis, 370 titrations, 212–13
molecular mechanics, 515 dilution factor, 230 essay questions, 558–9
quantum mechanics, 514–15 dilutions poor performance in, 560
sof tware for, 518 preparing, 75–6 essay writing, 594–6
concentration, in SI units, 43 series, 77 diagrams in, 595
condensers, 155 single, 75 Golden Rules, 595
confirmatory analysis, 472 sample analysis, 230 planning, 594–5
confounding variables, 48 dipole moment, 329 time management, 594
conical flasks, 64, 92 discontinuous variables, 36 evaporation
conical (Erlenmeyer) flasks, 67 dissertations, 597 gas ‘blow-down’, 163–4
consistency in measurement, 38 dissociation of acids, 113 techniques, 161
continuous variables, 36 dissociation of alkalis, 113 excited states, 217
continuous-wave (CW) spectrometers, 334 dissolution and recrystallisation, 132–4 experimental design, constraints, 48
Control of Substances Hazardous to Health distance learning, 549 exploratory data manipulation, 472
(COSHH), 9–13, 33 distillation exponential notation, 467
cooling chemicals, 93–5 equipment, 145–6 Extended Control of Substances Hazardous to
cooling probes, 94–5 types, 145 Health, 10–13
ice baths, 93 Dixon and Mood’s sign test, 494 extended matching items (EMI), 561
solid CO2 baths, 93–4 double-beam spectrometers, 320 extraction
cooling in recrystallisation, 136 double bond equivalents (DBE), 188–9 liquid–liquid, 139–40
cooling probes, 94–5 downloading files, 394 solid–liquid, 139, 142–3
cork rings, 83 drugs analysis, thermogravimetry on, 372 types, 139
correlation, 495–7 dry ashing of solids decomposition, 240
coulometric methods, 307 drying chemicals, 95–7 F
coupling (spin–spin splitting) in 1H@NMR dessicators for, 96–7 Fajans titrations, 213
spectra, 340–3 joint clips, 100–1 filtration, 83–7
Crystallography Open Database (COD), 409, jointed glassware for, 97–101 in gravimetry, 185–6
418–20 screwcap adaptors, 99–100 gravity, 83–4
crystals, collection of, 136–7 solvents, 97 of solids, 83, 86
CrystalWorks, 366 drying in recrystallisation, 137 suction, 84–6
current activities, analysing, 529–30 drying tubes for reflux, 156 flame absorption spectrophotometry, 222–3
curriculum vitae (CV) dynamic headspace (DHS) analysis, 284 flame atomic absorption spectrophotometry
adjustments to, 569 (FAAS), 226, 228–229
developing, 566–7 E operating, 233
enhancing, 569–71 ebook, 377 flame atomic emission spectrophotometry, 222–4
evidence-based, 566 ebrary, 377 flame ionisation detector (FID), 258
skills, 566 effective planning and working, tips for, 532 flame photometer, 233
structure and presentation, 567–9 eJournal, 377 flames tests in inorganic analysis, 182
cyclic voltammetry, 307 e-learning, 549 flasks
cylinders, 64 electrical field strength, 287 conical, 64
electric heated block for melting point greasing joints, 76, 98
D measurement, 125 round-bottom, heating, 93
data analyses, 562 electric heating mantles, 91 sealing, 76
database functions, 424 electroanalytical techniques, 302–7 volumetric, 64
databases, 432 electrochemical detection, 269 fluorescein, 221
data collection electrochemical methods, 302 fluorescence, 220–2
preparing a table for, 52 electrolysis, 305 fluorescence immunoassay (FIA), 222
628 Index

Index
fluorescence spectrophotometry, 221–2 greasing joints in glass apparatus, 77, 98 detectors, 235
fluorescent in situ hybridisation (FISH), 222 ground state, 217 interferences in, 235
formative assessments, 551 guessing, 559 sample introduction, 234
Fourier-transform (FW) spectrometers, spectrometers, 235
320–1 H industrial placements, 569
fractional distillation, 145–146 Handbook of Chemistry and Physics Online, inert atmosphere reactions
fractionating columns, 146 408–16 apparatus for, 167
frequency distributions, 482–3 Hartree–Fock method, 515 gas ‘bubblers’, 167
frequency polygons, 449 Harvard system of citations, 378 gas cylinders for, 166
full scan mode in LC-MS, 360 Harwood, L.M., 129 solvents for, 167–8
fusion of solids decomposition, 240 hazard, definition of, 6 syringe techniques, 168–70
hazard warning pictograms, 14 information
G headings evaluation sources of, 382–8
galvanic cell, 302 for poster display, 576 authorship and provenance, 384–5
gamma emission, 309–10 health and safety, 6–35 critical thinking, 386–7
gamma-ray spectrometry, 313 basic rules, 33–4 facts and ideas, 385–6
gas ‘blow-down’ evaporation, 163–4 legislation, 6 graphs, 386
gas burners, 87–8 risk assessment, 6–8 numerical data, 386
tubing for, 88 Health and Safety at Work Act 1974, 6 tables, 386
gas chromatography (GC) heating chemicals, 87–93 information-processing exams, 561–2
components, 255 bumping, 87 infrared spectroscopy, 318–28
detectors, 258–259 burners for, 87–8 absorption bands, 319
mass spectrometry in, 357 electric heating mantles, 91 spectra, 319–20
sample injection, 255 hot air guns, 92 interpretation of, 325–8
gas–liquid separator, 227–228 hot glassware, handling, 92–3 spectrometers, 320–1
Gaussian distributions, 490–1 oil baths, 90–1 inorganic analysis, quantitative techniques
Gaussian package, 515 steam and water baths, 88–9 for, 179–82
Geiger–Müller (G–M) tube, 311 stirrer hot plates, 90 anions and cations, testing for, 179–80
gel concentration, 292–3 heating too fast, 126 centrifugation of solutions, 180–1
gel permeation chromatography Henderson-Hasselbalch equation, 117 flames tests, 182
(GPC), 251–2 hexaammine-cobaltate, structure of, 501 heating test tubes, 181–2
glass apparatus high resolution mass spectra (HRMS), 188 reagents, 179
containers, 67–8 Hirsch funnels, 84–5 typical reagents, 179
cleaning, 68 histograms, 472–473 Inorganic Crystal Structure Database (ICSD), 366
1 insolubility, 106
drying, 95 H@NMR spectra, 335–44
greasing joints in, 77, 98 chemical shift, 333 integration in 1H@NMR spectra, 339–40
jointed, 97–101 correlation tables, 337–9 interconversion of SI units, 44
care of, 98–9 coupling, 340–3 interferences
safety with, 68–9 integration of peak areas, 339–40 in inductively coupled plasma AES, 235
glass membrane electrodes, 304 hollow-cathode lamp (HCL), 226–227 spectral
glass pipettes, 64 Hooke’s law, 318 in AAS, 232
glucose electrode, 307 hot air guns, 92 International Center Diffraction Data
Golden Rules for essay writing, 595 hot filtration in recrystallisation, 134–5 (ICDD), 366
gradient elution separation in HPLC, 265 hot plates, 90 internet
graduate attributes, 527 stirrer, 90 browsers, 392–3
graphics and presentation packages, 433 H (hazard) statements, 8, 13, 15–19 directories, 394
graphite furnace atomiser, 227–228 hydrogen bonding, 129 resources for chemistry, 399–420
graphs, 447–57 Hyperchem program, 515 databases, 401–19
interpretation of, 455 principal resources, 400
misrepresentation with, 456–7 I search tools, 393–4
practical aspects, 447–9 ice baths, 93 interpersonal skills, 534
checklist, 450 ideal/non-ideal behaviour of solutions, 102 interval scale, 37
large or small numbers, 448 immunohistochemistry, 222 interviews for assessment, 562–3
paper for, 448–9 indicators ion-exchange chromatography (IEC), 251
size, 448 for acid–base titrations, 199 ionic structures, drawing, 503
types, 449 in complexometric titrations, 205 ionisation interferences in AAS, 232
gravimetry, 184–6 induced dipole moment, 329 ionophore, 305
analytical procedure, 184 inductively coupled plasma mass spectrometry, ion-selective electrodes (ISEs), 303–4
filtration, 185–6 235–8 ion trap mass spectrometer, 359
precipitation, 185 collision-reaction cell, 237–8 isobaric interferences in ICP-MS, 236–7
gravity chromatography, 263 interferences, 236–7 isoelectric focusing (IEF), 296
gravity filtration, 83–4 schematic diagram, 236 isotope dilution analysis (IDA), 236
flute filter for, 84 inductively coupled plasma spectroscopy, 234–5 isotopes, peaks in MS, 354
Index 629

Index
J M mull infrared spectrum, 321–3

joint clips, 100–1 Mann–Whitney U-test, 494 multi-factorial experiments, 50
junk mail, 391 MassBank, 408, 416–417 multiple-choice questions (MCQs),
mass in SI units, 43 559–61
mass spectrometry, 318
K
‘Katritzky’ system, 378–9
base peak, 354 N
detectors for, 359 National Chemical Database Service, 403
KBr disk
fragmentation patterns, 353–6 neutralisation curves in acid–base titrations,
infrared spectrum of, 321–322
full scan mode, 360 199–200
preparation of, 324
in gas chromatography, 357 nominal scale, 37
Kelvin scale, 37
mass spectra, 353–4 normality of solutions, 106
Kofler block, 123–124
molecular ions, 354 note-taking, 538–42
Kolmogorov–Smirnov test, 494
sample handling, 353 from books and journals, 540–2
Kruskal–Wallis H test, 498
types of, 358–9 in lectures, 538–40
Kugelrohr distillation, 151
Materials Safety Data Sheets (MSDS), making up, 540
kurtosis in frequency distributions, 483
8, 25–32 scanning, 540–1
mathematical models, 47 skimming, 541–2
L mean, 476–7 nuclear magnetic resonance (NMR), 318
laboratory environment, working measurements, 36–9 chemical shift, 333
in a, 58 accuracy, 38 13
C@NMR spectra, 343–6
laboratory procedures, 70–80 bias, 38 1
H@NMR spectra, 335–44
balances and weighing, 78–80 error, 38–9 sample handling, 334
chemicals, using, 70 precision, 38 sample preparation, 335
clamps and support stands, 82–3 scales, 37 spectra, interpretation of, 335–49
cooling chemicals, 93–5 variables, 36 spectrometers, 333–5
dilutions, preparing, 75–6 median, 477, 479 spectroscopy, 332–50
drying chemicals, 95–7 melting point depression, 123 nuisance variables, 48
filtration, 83–7 melting points, 123–7 null hypothesis, 487–8
heating chemicals, 87–93 apparatus for, 123–5 numerical problems, solving, 463–70
solutions, preparing, 71–5 capillary tubes, 124–5 exponents, 467
laboratory work, basic rules for, 33–4 dertermination of, 125–6 linear functions, 468–9
lab partnerships, 536 electric heated block for, 125 logarithms, 468
layout for poster display, 575–6 heating too fast, 126 percentages and proportions, 467
learning objectives, 551 Kofler block for, 123–124 rounding to decimal places, 465–7
least significant difference, 494 mixed melting point dertermination, 127 scientific notation, 467–8
lectures, 546–8 oil bath apparatus for, 125 steps in, 464–5
Lewis structures, drawing, 502 purity, criterion of, 123
ligand type titrations, 203 Merck Index Online, 404–5
light absorption, principles of, 217–20 metal-ion indicators, 205
O
Office suite for spreadsheets, 421
light sensitive chemicals, storing, 67 methane, 501
oil bath apparatus for melting point
liquid chromatography, 260–9 method of standard additions, 229
measurement, 125
liquid chromatography-mass spectrometry micellar electrokinetic chromatography
oil baths, 90–1
(LC-MS), 358 (MEKC), 298
online communication, 389–92
full scan mode, 360 microburners, 88
online material, 380
selected ion monitoring (SIM) Microsoft PowerPoint®, 5
online sources of information, 389–98
mode, 360 Miller indices, 366, 368
internet as source, 389
liquid–liquid extraction, 139–40 mixed melting point dertermination, 127
oral exams, 562–3
liquid–liquid partition mixed solvent recrystallisation, 129–130
ordinal scale, 37
chromatography, 250 procedure, 135
organic structures, drawing, 501–2
liquids, 63–9 selection for recrystallisation of unknown
organising numbers, 472–3
glass and plastic containers, 67–8 compound, 132
osmotic effects, 102
holding and storing, 65, 67 Mohr titrations, 212–13
oxidation, 302
infrared spectrum of, 321–3 molality, 103
oxidizing agents in titrations, 208
measuring and dispensing, 63–5 molarity, 102
specialised apparatus, 67 Molden program, 515
liquid scintillation counting, 314 Moldraw program, 515 P
literature surveys, writing, 606–8 mole, 102 parallel synthesis in combinatorial
defining terms, 607 molecular dynamics, 515 chemistry, 174–5
opposing views, balancing, 607 molecular formulae, 187–9 paraphrasing, 541
organisation of, 606 molecular interferences in ICP-MS, 236–7 partition chromatography, 250–1
structure and content, 607–8 molecular mechanics, 514 partition coefficients in liquid–liquid
time management in, 606 molecular orbital diagrams, 504–5 extraction, 140
topics, 606 monochromator, 218 Pasteur pipettes, 63–4
630 Index

Index
Pearson’s product moment correlation, 496 procedure dissolution, 132–4

peer assessment, 534 redox, 208–10 drying, 137
Periodic Table database, 401–2 proficiency testing schemes, 438 hot filtration, 134–5
personal communications, 380 project proposal, 57 problems in, 137
personal development planning (PDP), project work process of, 130–7
525–7 recording details of, 53 single-solvent recrystallisation, 133
and curriculum vitae (CV), 566 proportional counter, 362 solvents for, 128–9
personal reference library in scientific P (precautionary) statements, 8–9, 20–4 yields from, 129
writing, 592 reduced pressure distillation, 145, 148–9
pH, 113–14 reduction, 302
electrodes, 114–16
Q titrations
quadruple mass spectrometer, 359
indicator dyes, 114 in volumetric analysis, 190
qualitative analysis, 437
measurement of, 114–16 reduction–oxidation (redox) reactions, 108–9
qualitative variables, 36
physical interferences in AAS, 232 titrations, 208–10
pipette fillers, 64 reflux
aqueous solutions in, 74
pipettes, 64 with addition of chemicals, 157–8
atomic spectroscopy, 222–4, 226–40
in volumetric analysis apparatus, setting up of, 155–6
definition, 437
filling, 196 with mechanical stirring, 157–9
fluorescence, 220–2
using, 193, 195 regression, 495–7
fluorescence spectrophotometry, 221–2
pipettors, 64–5 relational database, 432
selection, criteria for, 439
delivering accurates volumes, 66 relative atomic mass, 102
spectroscopic techniques, 217–24
plastic containers, 67–8 research papers, obtaining and
validity and, 437–8
polarizability, 329 organising, 377
weighing solid samples, 80
polymer degradation, thermogravimetry resonance Raman spectroscopy (RRS), 329
quantitative techniques for inorganic
on, 372 revision, 552–5
analysis, 179–82
positive displacement pipettors, 65 in scientific writing, 592
anions and cations, testing for, 179–80
poster display risk assessment
centrifugation of solutions, 180–1
content, 578 important aspects, 6
flames tests, 182
design, 575–7 matrix analysis, 8
heating test tubes, 181–2
poster session, 578, 580 process, 6–7
reagents, 179
preliminaries, 575 risk, definition of, 6
typical reagents, 179
potassium hydrogen phthalate, 117 rotary film evaporation, 161–3
quantitative variables, 36
potentiometry, 303–5 rovaps, 161
quantum mechanics, 514
PowerPoint Royal Society of Chemistry databases, 402–3
question-spotting, 554
for poster display, 579
quoting, 541
for spoken presentations, 582 S
practical and project work, 597–604 saturated solutions, 106
advanced, 4–5 R scales of measurement, 37
approaches to, 3–5 radioactive isotopes scanning, 540–1
calculators, 4 chemical applications for, 314–15 Science Citation Index (SCI), 376
preparation, 3 radioactive decay, 309 scientific notation, 467–8
requirements, 3–5 working practices when using, 315–17 scientific paper, producing, 604
results, recording, 3–4 Radioactive Substances Act (1993), 315 scientific reports
scientific paper, producing, 604 Raman spectroscopy structure of, 59–60
steps in, 599 advantages, 330 scientific writing, 587–93
structure of, 598 energy level diagram, 328 common errors, 592–3
theses and dissertations, 597 principle, 329 organising information and ideas, 587–9
practical laboratory reports, 597 schematic diagram, 329 revision, 592
practicals, 547–8 water molecule, 329 spider diagrams in, 588
precipitation range, 480 time management, 587
in gravimetry, 185 ranked variables, 36 scintillation counter, 311–312
reactions, 108 ratio scale, 37 scintillation screen, 362
titrations, 211–13 reaction flasks, 155 scratching for recrystallisation, 136
end-point determination, 212–13 reactions of ions in solution, 107–9 screwcap adaptors, 99–100
in volumetric analysis, 190 acid–base reactions, 107–8 secondary record of research, 53–4
precision in quantitative analysis, 38 complexation reactions, 108 selected ion monitoring (SIM) mode in
press and pull method, 364 precipitation reactions, 108 LC-MS, 360
pressurised fluid extraction (PFE) reduction–oxidation (redox) reactions, 108–9 semi-conductor, 362–3
system, 277 Really Simple Syndication (RSS), 399 semi-empirical method, 514
pressurised microwave-assisted extraction recrystallisation, 128–37 semi-interquartile range, 480–1
system, 276 collection of crystals, 136–7 separatory funnels, 140–2
primary record of research, 53 cooling, 136 short-answer questions (SAQs), 559–61
principal component analysis, 511–12 decolorisation, 134 side-loading method, 365
Index 631

Index
simple distillation, 145–146 selected properties, 129 summative exams, 557–8

single-pan balance, 78 selection, 130 support rings, 82–3
sintered-glass funnels, 87 for recrystallisation of unknown support stands, 82, 84
SI units, 40–4 compound, 131 surface enhanced Raman spectroscopy
basic format, 41 single-solvent recrystallisation, 133 (SERS), 329
in chemistry, 43–4 Soxhlet extraction, 143 surface enhanced resonance Raman spectroscopy
conversion factors, 42 spatula, 179 (SERRS), 329
and derived units, 40–41 specialised information, 376 syringes, 65, 168–70
describing measurements in, 41–3 specific activity of isotopes, 311–312
interconversion of, 44 spectral interferences
physical constants in, 43 in AAS, 232 T
prefixes, 40–41 spectrometry, 318 tables, 459–62
skills, in curriculum vitae (CV), 566 spectroscopic techniques, 217–24 preparation of, 459–60
skimming, 541–2 atomic spectroscopy, 222–24 team
sodium hydroxide solutions fluorescence, 220–2 definition, 534
standardisation of, 200–2 UV/visible spectrophotometry, 217–20 role, 534
using KPH, 201–2 spider diagrams teamworking
soil analysis, thermogravimetry on, 372 in scientific writing, 588 dynamics, 535–6
solid–liquid extraction, 139, 142–3 spoken presentations, 581–5 skills, 534–5
solid-phase microextraction (SPME), 278–84 audio-visual aids for, 581, 583 temperature in SI units, 44
solid phase synthesis, 173–4 content, 583–4 test tubes, 65
solids hints on, 585 heating, 88, 181–2
decomposition techniques, 238–40 PowerPoint for, 582 holding, 92
acid digestion, 238–40 preparation, 581, 583 text
dry ashing, 240 spreadsheets for poster display, 576
fusion, 240 constructing, 425 theory, 46
drying, 95–6 data entry, 421–2 thermal analysis, 370–2
infrared spectrum of, 321–3 parts of, 423–4 thermal conductivity detector (TCD), 258
purity of, 123 printing, 425 thermogravimetry, 370–1
suction filtration of, 86 templates, 424–5 interpretation of trace, 371
solid-state membrane electrodes, 305 spreadsheet statistics functions, 433 thermometers, 123
solubility, 106–7 SQ3R technique for skimming, 542 calibration of, 126
product, 106–7 standard solutions, 73 theses, 597
storing, 76–77 in quantitative analysis time in SI units, 43
solutions, 102–11 in given concentration range, 341 time management
analytical in titrations, 192–3 advantages, 529
preparation, 71, 73–4 static headspace (SHS) analysis, 284 definition, 529
procedures, 75 statistical analysis packages, 432–3 in essay writing, 594
aqueous, 72, 74 statistical tests, 487–99 quality in, 530
centrifugation of, 180–1 choosing, 491–7 in scientific writing, 587
concentration, 102–6 confidence limits, 495 when revising, 552
activity, 104–5 correlation and regression, 495–7 in writing literature surveys, 606
equivalent mass, 105 dispersion, comparing, 494 time-of-flight mass spectrometer, 359
molality, 103 frequency distributions, 494 time-wasting activities, 530–1
molarity, 102 location, comparing, 491, 494 titles
normality, 106 proportions, 494 for poster display, 576
parts per million (ppm), 104 computers, use of, 497–9 titrations, 192–3
per cent composition (% w/v and % null hypothesis in, 487–8 acid–base, 199–202
v/v), 103–4 parametric distributions, 488–91 calculations, 208–10
per cent composition (% w/w), 103 steam baths, 88–9 end-point determination, 212–13
percentage concentration, 71 steam distillation, 145, 151 measurement by, 190
preparing, 71–5 stem and leaf plots, 473 precipitation, 211–3
reactions of ions, 107–9 stirrer hot plates, 90 precision in, 195–6
acid–base reactions, 107–8 stock solution, 73 procedure, 195
complexation reactions, 108 of metal ions, 226 standard solutions, 192–3
precipitation reactions, 108 Stokes shift, 221 transcripts, 556
reduction–oxidation (redox) reactions, Student’s t test, 491–3 transferable skills, 523–5
108–9 subtitles for poster display, 576 in chemistry, 524–5
of sodium hydroxide, 200–2 suction filtration, 84–6 transformations, 473–5
solvents of solids, 86 transmittance, 218
drying, 97 summarising, 541 Turbomole package, 515
for recrystallisation, 128–9 summative assessments, 551 tutorial groups, 536–7
632 Index

Index
tutorials, 548–9 Volhard titrations, 212–13 word processors, 428–9

two-dimensional electrophoresis, 296–7 voltammetric methods, 305–6 editing features, 429–30
two-level factorial design, 509 volume in SI units, 43 fonts and line spacing, 430–1
volumetric analysis, 190–7 printing, 431–2
burette, filling, 193 table construction, 431
U calculating concentrations, 197 World Wide Web (WWW), 399
Ugi reaction in combinatorial chemistry,
calculations used, 191–2
174–5
pipette
unit cells, 362, 365
filling, 196
X
urgent tasks, 531 X-ray diffraction (XRD), 362–8
using, 193, 195
UV/visible spectrophotometer, 218–19 detectors, 362–3
reaction types, 190
types of, 219 procedure, 363–8
titrations, 192–3
use of, 220 X-ray fluorescence spectrometry
measurement by, 190
UV/visible spectrophotometry, 217–20 (XRF)
precision in, 195–6
light absorption, principles, 217–20 detectors, 244
procedure, 195
quantitative analysis with, 219–20 environmental application, 248
standard solutions, 192–3
evaluating, 244–6
volumetric flasks, 64
instrumentation, 243–4
V sample preparation
vacuum dessicators, 96 W for liquid samples, 246–7
vacuum distillation, 145, 148–9 water baths, 88–9 for solid samples, 247–8
valence shell electron pair repulsion (VSEPR) wavelength dispersive X-ray fluorescence X-rays
theory, 504 (WDXRF), 243 generation, 366
validation in quantitative analysis, 437–8 web-based resources, 535 nomenclature, 366
Vancouver system of citations, 378 weighing
variables, 36 containers, 79–80
VAK system of learning, 546 solid samples, 80 Z
viscous liquids, transferring, 163 Wilcoxon’s signed rank test, 494 zwitterionic buffers, 117
Index 633


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Practical Skills in Chemistry John R. Dean • Alan M.

Jones • David Holmes

This edition provides:

Practical Skills in Chemistry, 3rd edition, is an indispensable book for undergraduate

Reed • Weyers • Jones

Cover image: Michael Rozewski/Getty Images www.pearson-books.com

CVR_DEAN_03_39920.indd 1 02/05/2017 14:01

A01 Practical Skills in Chemistry 39920.indd 1 11/05/2017 09:13

We combine innovative learning technology with trusted

From classroom to boardroom, our curriculum materials, digital

Every day our work helps learning flourish, and

To learn more, please visit us at www.pearson.com/uk

A01 Practical Skills in Chemistry 39920.indd 2 11/05/2017 09:13

A01 Practical Skills in Chemistry 39920.indd 3 11/05/2017 09:13

First published 2002 (print)

© Pearson Education Limited 2002 (print)

ISBN: 978-1-292-13992-0 (print)

British Library Cataloguing-in-Publication Data

Library of Congress Cataloging-in-Publication Data

Print edition typeset in 10/12pt Times LT Pro by Spi Global

A01 Practical Skills in Chemistry 39920.indd 4 11/05/2017 09:13

List of boxes viii

The investigative approach 1

Fundamental laboratory techniques 61

Laboratory techniques 121

Classical techniques 177

A01 Practical Skills in Chemistry 39920.indd 5 11/05/2017 09:13

Instrumental techniques 215

Information technology and library resources 373

Analysis and presentation of data 435

Study and examination skills 521

A01 Practical Skills in Chemistry 39920.indd 6 11/05/2017 09:13

Communicating information 573

Answers to study exercises 609

A01 Practical Skills in Chemistry 39920.indd 7 11/05/2017 09:13

2.1 How to perform a risk matrix analysis 8

A01 Practical Skills in Chemistry 39920.indd 8 11/05/2017 09:13

30.5 Analysis of a sample: dilution factor 230

A01 Practical Skills in Chemistry 39920.indd 9 11/05/2017 09:13

5 1.1 Example of using the algebraic rules of Table 51.2 465

A01 Practical Skills in Chemistry 39920.indd 10 11/05/2017 09:13

A01 Practical Skills in Chemistry 39920.indd 11 11/05/2017 09:13

43 Using online resources

Information and communication technology (ICT) is vital in the modern aca-

Definitions of key terms and

ful hints and practical advice,

and are highlighted in the text Understanding the technology – you

features of methodology. wireless network can be complex, but

the hardware. White and Downs (2014) online communication

Information technology and library resources 389

M43 Practical Skills in Chemistry 39920.indd 389 10/05/2017 20:22

Basic laboratory procedures I

Instrumental techniques 309

M09 Practical Skills in Chemistry 39920.indd 80 10/05/2017 16:32

A01 Practical Skills in Chemistry 39920.indd 12 11/05/2017 09:13

Figures are used to illustrate (a) (b)

95% dimethylsiloxane–5% diphenylypolysiloxane:

(c) (d) 5% 95%

Fig. 32.12 Schematic diagram of a GC column.

H251 Self-heating; may catch fire Self-heating sub- Category 1 Danger