1.

UV-Visible spectroscopy
The Electromagnetic Spectrum
The visible spectrum constitutes a small part of the total radiation spectrum. Most of the
radiation that surrounds us cannot be seen, but can be detected by dedicated sensing instruments.
This electromagnetic spectrum ranges from very short wavelengths (including gamma and x-
rays) to very long wavelengths (including microwaves and broadcast radio waves). The
following chart displays many of the important regions of this spectrum, and demonstrates the
inverse relationship between wavelength and frequency (shown in the top equation below the
chart).

The energy associated with a given segment of the spectrum is proportional to its
frequency. The equation below describes this relationship, which provides the energy carried by
a photon of a given wavelength of radiation.

Electronic Transition: (Self Study)

The visible region of the spectrum comprises photon energies of 36 to 72 kcal / mole, and
the near ultraviolet region, out to 200 nm, extends this energy range to 143 kcal / mole.
Ultraviolet radiation having wavelengths less than 200 nm is difficult to handle, and is seldom
used as a routine tool for structural analysis.
The energies noted above are sufficient to promote or excite a molecular electron to a
higher energy orbital. Consequently, absorption spectroscopy carried out in this region is
sometimes called "electronic spectroscopy".
 A diagram showing the various kinds of electronic excitation that may occur in organic
molecules is shown above. Of the six transitions outlined, only the two lowest energy ones (left-
most, colored blue) are achieved by the energies available in the 200 to 800 nm spectrum. As a
rule, energetically favored electron promotion will be from the highest occupied molecular
orbital (HOMO) to the lowest unoccupied molecular orbital (LUMO), and the resulting species is
called an excited state.
When sample molecules are exposed to light having an energy that matches a possible
electronic transition within the molecule, some of the light energy will be absorbed as the
electron is promoted to a higher energy orbital. An optical spectrometer records the wavelengths
at which absorption occurs, together with the degree of absorption at each wavelength. The
resulting spectrum is presented as a graph of absorbance (A) versus wavelength, as in the
isoprene spectrum shown below. Since isoprene is colorless, it does not absorb in the visible part
of the spectrum and this region is not displayed on the graph. Absorbance usually ranges from 0
(no absorption) to 2 (99% absorption), and is precisely defined in context with spectrometer
operation.

When a beam of light passes through an absorbing medium, for example a solution, a part
of the light is absorbed and the rest is transmitted. The amount of light absorbed depends on the
concentration of the solution and the length traversed by the light through the solution. The
quantitative relation between the amount of light absorbed, concentration and the length of the
absorbing medium is governed by the laws- the Lambert’s Law, the Beer’s Law.

Lambert’s Law:
The law states that “Equal fractions of the incident light are absorbed by successive layers
of equal thickness of the absorbing medium”.
If a monochromatic light passes through a transparent medium, the rate of decrease in intensity
with the thickness of medium is proportional to intensity of the incident light i.e.
-dI/I α dx
dI/I = - k1 dx-------(i)
Where I is intensity of the incident light of wavelength λ, x is the thickness of the medium and k1
is proportionality factor. The negative sign indicates that due to absorption, the intensity of light
decreases as it passes through the absorbing medium.
If I0 is the initial intensity of incident light on the absorbing medium of length l and It is the
intensity of transmitted beam, then the integration of eqn.(i) becomes
ln(It / Io) = - k1 . l
ln(Io/ It) = k1 . l
log10(Io/ It) = k1 x l
2.303-----------(ii)
Beer’s Law:
The law states that “Equal fractions of the incident light are absorbed by successive layers
having equal concentration of the absorbing medium”.
The intensity of a beam of monochromatic light decreases exponentially as the
concentration of the absorbing substance increases arithmetically. i.e.
-dI/I α dc
dI/I = - k2 dc ------(iii)
If I0 is the initial intensity of incident light on the absorbing medium of concentration c and It is
the intensity of transmitted beam, then the integration of eqn.(iii) becomes
ln(It / Io) = - k2 . c
ln(Io/ It) = k2 . c
log10(Io/ It) = k2 x c
2.303-----------(iv)

Beer-Lambert’s law:
A combination of Lambert’s Law and Beer’s Law results in Beer-Lambert’s Law, it states
that “Equal fractions of the incident light are absorbed by successive layers of equal
thickness and equal concentration of the absorbing medium.”
Combining (ii) and (iv),
log10(Io/ It) = k2 x c x l
2.303
log10(Io/It) =  c l
Log(Io/It) is called as absorbance, if c is expressed in mol dm-3 and l in cm then  is called as
molar absorptivity or absorption coefficient.
Hence, A =  c l = log(Io/It) = log (1/T) = -log(T) where T is transmittance.

Conditions: The law is only true for monochromatic light, which is light of a single wavelength
or narrow band of wavelengths, and provided that the physical or chemical state of the substance
does not change with concentration.
 Io is the intensity of the incident radiation and I is the intensity of the transmitted
radiation. The ratio I/Io is called transmittance.This is sometimes expressed as a percentage and
referred to as %transmittance.
Mathematically, absorbance is related to percentage transmittance T by the expression:
A = log10(Io/I) = log10(100/T) =  x c x L
Where, L is the length of the radiation path through the sample, c is the concentration of
absorbing molecules in that path, and is the molar extinction coefficient - a constant dependent
only on the nature of the molecule and the wavelength of the radiation. If concentration is
expressed in mol/dm3 and L is expressed in dm then unit for is dm2/mol.

Limitations of the Beer-Lambert law

1. The linear relationship between absorbance and concentration of solution is not observed
at concentration above 10-2 M, hence concentrated solution do not obey Beer-Lambert’s
Law.
2. The law does not obeyed if the absorbing species reacts with the solvent, dissociates or
associates in solution. The molecules of the absorbing species should remain as simple
molecules and should not undergo any change in molecular condition.
3. The light incident on the absorbing medium should be monochromatic otherwise minor
deviations from Beer-Lambert’s Law are observed. Hence monochromators have to be
used to produce monochromatic beams.
 4. The molar absorptivity depends on the refractive index of the absorbing medium. The
refractive index changes with the concentration of the absorbing medium. At high
concentration these changes are considerable but at concentrations below 10-2 M, these
changes can be neglected.
5. Temperature fluctuations and entry of stray light into the absorbing system also lead to
deviations from Beer-Lambert’s law.
6. The Fluoresecenceor phosphorescence can cause a positive deviation in % T and
negative deviation for A in the system.
7. Non-monochromatic radiation, deviations can be minimized by using a relatively flat part
of the absorption spectrum such as the maximum of an absorption band.

Ultraviolet-visible spectrophotometer
The instrument used in ultraviolet-visible spectroscopy is called a UV/Vis spectrophotometer.It
measures the intensity of light passing through a sample (I), and compares it to the intensity of
light before it passes through the sample (I_o). The ratio I/I_ois called the transmittance, and is
usually expressed as a percentage (%T). The absorbance, A, is based on the transmittance:

The UV-visible spectrophotometer can also be configured to measure reflectance. In this case,
the spectrophotometer measures the intensity of light reflected from a sample (I), and compares
it to the intensity of light reflected from a reference material (Io) (such as a white tile). The ratio
I/Io is called the reflectance, and is usually expressed as a percentage (%R).
The basic parts of a spectrophotometer are a light source, a holder for the sample, a
diffraction grating in a monochromator or a prism to separate the different wavelengths of light,
and a detector.
Single beam spectrophotometer
A single beam spectrophotometer is comprised of a light source, a monochromator, a
sample holder, and a detector. An ideal instrument has a light source that emits with equal
intensity at all wavelengths, a monochromator that is equally efficient in splitting light into
narrow groups of wavelength for all wavelengths, and a detector that is sensitive and responds
equally to all wavelengths.
Light sources
Because no single light source with the appropriate characteristics exists,
mostspectrophotometers use two lamps, with one for the ultraviolet region and one for the visible
region. The visible lamp is usually a tungsten lamp(300-2500 nm), while the ultravioletlamp is a
deuterium lamp(190-400 nm). An alternative, relatively rarely used inspectrophotometers,
although commonly used in other types of spectroscopic instruments, is a xenon arc lamp (160-
2,000 nm).
Monochromators
Although prisms can be used as monochromators, most instruments use
diffractiongratings. Light shining on the closely spaced grooves of a diffraction grating at an
angle is separated into different wavelengths in a consistent manner, assuming that the grooves
are consistently produced.
Detector
The detector is typically a photomultiplier tube, a photodiode, a photodiode array or a
charge-coupled device (CCD). Single photodiode detectors and photomultiplier tubes are used
with scanning monochromators, which filter the light so that only light of a single wavelength
reaches the detector at one time. The scanning monochromator moves the diffraction grating to
"step-through" each wavelength so that its intensity may be measured as a function of
wavelength. Fixed monochromators are used with CCDs and photodiode arrays. As both of these
devices consist of many detectors grouped into one or two dimensional arrays, they are able to
collect light of different wavelengths on different pixels or groups of pixels simultaneously.
The most commonly used detector is a photomultiplier tube (PMT). An incomingphoton
hits a thin metal film inside a vacuum tube. The metal film is maintained at a large negative
potential, and emits electrons. These collide with a series of dynodes maintained at progressively
lower potentials; each dynode emits several electrons in response to each incoming electron,
resulting in a large amplification of the signal. Because the initial photon is required to initiate
the process, most PMTs have very little dark current (“dark current” is signal without light).
Proper functioning of a PMT requires a constant voltage across the PMT; maintaining a constant
voltage in the face of a high signal requires a well-designed instrument. PMTs are wavelength
dependent, with the degree of dependence being related to the metal used in the thin film; most
PMTs exhibit the greatest sensitivity at ~400 nm.
An alternative type of detector uses photodiodes. Photodiodes are inexpensive but not
very sensitive. Their low cost has allowed arrays of photodiodes to be set up to allow
simultaneous detection of many wavelengths. In this type of spectrophotometer, the
monochromator is located after the sample, so that it splits the multiwavelength light leaving the
sample.
A charge coupled device (CCD) is a sensitive array detector. CCDs store charges released
in response to photon impacts. Because the stored charges are stable for prolonged periods, a
CCD can collect data for considerable time prior to readout of the signal. They are therefore
potentially extremely sensitive. They will probably displace PMTs from some uses as their price
decreases. CCDs are used in digital cameras and other consumer products and are rapidly
becoming less expensive as a result of both economies of scale and the development of improved
production techniques.
Cuvettes (Sample Holder)
Most samples studied using visible and ultraviolet spectroscopy is liquid. Thesample
must therefore be placed in a transparent container to allow measurement.These containers are
called cuvettes. Cuvettes are generally made from transparent plastic, glass, or quartz.
Differentcuvettes have different optical properties.
Working: The sample holder is filled with the solvent (blank). The absorbance value of the
solvent is adjusted to zero for a particular wavelength, obtained by the rotation of the
monochromator. The sample is then taken in the sample holder and its absorbance value is
determined for the same wavelength. The procedure is repeated for different wavelengths
obtained by the rotation of the monochromator. The absorbance values can be read either on a
dial or a digital display. The λmax values for the sample can be thus found.
Double Beam Spectrophotometer
A beam of visible light from an incandescent tungsten lamp passes through a colour filter
which selects a narrow band of wavelengths. A mirror splits this narrow band into two beams-
one passing through the sample and the other passing through the solvent(blank) hence the name
Double beam Spectrophotometer.
The sample absorbs a part of beam whereas the solvent transmits it completely. The two
beams then fall on the respective photocells, where photoelectrons are emitted and recorded.

Working: The solvent is first taken in both cuvettes, and zero level adjusted. The sample is then
placed in the sample cuvette. It absorbs a part of the light and the transmitted beam emerging
from it falls on the photodiode. This beam has less intensity than when only solvent was present
in the sample cuvette. Hence there will be proportionate decrease in the electric current produced
in photodiode and recorded.
Advantages of Double beam spectrophotometer over single beam spectrophotometer:
 1. Changes in the intensity pf incident light due voltage fluctuations in the power supply are
compensated by splitting the incident beam into identical beams by the use of a mirror
and two balanced photodiodes. Any error due to solvent or impurity is balanced out as
identical beams pass through the blank and sample as the absorbance of blank is initially
adjusted to zero.
2. The readings are not affected by changes in sensitivity of photodiodes as the zero method
is used.
3. The scale of instrument is linear with the concentration of the sample solution.

Applications:
1) Determination of λmax and identifying the functional groups: It is possible to
identify a particular group in a molecule by determining its λmax value. In
spectrophotometers, the beams of characteristic wavelength are produced, these are
absorbed by sample, which are indicative of functional groups as different functional
groups in the molecule absorb their own λmax values.
Function group Example λmax in nm Solvent
-COOH Acetic acid 208 Ethyl alcohol
-COCl Acetyl chloride 220 Hexane
-CONH2 Acetamide 178 Hexane
-NO2 Nitromethane 201 Methyl alcohol

2) Quantitative Analysis (Calibration curve method): UV/Vis spectroscopy is

routinely used in analytical chemistry for the quantitative determination of different
analytes, such as transition metal ions, highly conjugated organic compounds, and
biological macromolecules. Spectroscopic analysis is commonly carried out in
solutions but solids and gases may also be studied.
In this, a series of solutions of different concentrations of the compound is prepared.
Usually the concentrations vary from 10-3 M to 10-2 M. The absorbance is then
determined for these standard solutions at the wavelength which absorbed maximally
by the coloured compound (λmax). A graph of absorbance versus the concentration
is then plotted.
 From the calibration curve the concentration of the unknown solution can be found
out as shown.
3) Photometric titrations: Spectrophotometric measurements can be employed to
advantage in locating the equivalence point of a titration, provided the analyte,
reagent or titration product absorbs radiation. Alternatively absorbing indicator can
provide the absorbance change necessary for the location of equivalence point.
4) A UV/Vis spectrophotometer may be used as a detector for HPLC. The presence of
an analyte gives a response assumed to be proportional to the concentration. For
accurate results, the instrument's response to the analyte in the unknown should be
compared with the response to a standard; this is very similar to the use of calibration
curves. The response (e.g., peak height) for a particular concentration is known as the
response factor.
5) UV-Vis spectroscopy is also used in the semiconductor industry to measure the
thickness and optical properties of thin films on a wafer. UV-Vis spectrometers are
used to measure the reflectance of light, and can be analyzed via the Forouhi-Bloomer
dispersion equations to determine the Index of Refraction (n) and the Extinction
Coefficient (k) of a given film across the measured spectral range.
 2. pH Metry

Definition of pH
pH is an abbreviation of “pondus hydrogenii” and was proposed by the Danish scientist
S.P.L. Sørensen in 1909 in order to express the very small concentrations of hydrogen ions.
In 1909, pH was defined as the negative base 10 logarithm of the hydrogenion
concentration. However, as most chemical and biological reactions are governed by the hydrogen
ion activity, the definition was quickly changed. As a matter of fact, the first potentiometric
methods used actually resulted in measurements of ion activity.
The definition based on hydrogen ion activity is the definition we use today:
pH = - log10aH+

pH Theory:
pH is measured using a setup with two electrodes: the indicator electrode and the
reference electrode. These two electrodes are often combined into one - a combined electrode.
When the two electrodes are immersed in a solution, a small galvanic cell is established. The
potential developed is dependent on both electrodes. Ideal measuring conditions exist when only
the potential of the indicator electrode changes in response to varying pH, while the potential of
the reference electrode remains constant.
The measured voltage can be expressed by the Nernst equation in thefollowing way:
E = Eind - Eref = E'T - R • T/F • lnaH+
where
E = Measured voltage (mV)
Eind = Voltage of indicator electrode (mV)
Eref = Voltage of reference electrode (mV)
E'T = Temperature dependent constant (mV)
R = Gas Constant (8.3144 J/K)
T = Absolute Temperature (K)
F = Faraday's constant (96485 C)

By using the base ten logarithm, the formula can be written as:
E = E'T - 2.303 • R • T/F • log aH+
By introducing the pH definition as pH = -log aH+, pH can be expressed at the temperature T as
follows:
pHT = pHT° - ____E_____
R' • S • T
where
R' = constant = 0.1984 mV/K
S = sensitivity, a correction factor which takes into account that the electrode response may
differ from the theoretical value.
pH° = zero pH which is defined as the pH value at which the measured potential is zero. Figure 2
illustrates that the pH° will change with temperature and that another slope will be observed.

Working of pH- meter

A pH meter measures the potential difference (in mV) between the electrodes and
converts it to a display of pH.
In order to obtain a correct measurement, the input amplifier and theconverting circuit
must meet certain requirements. The principal construction of a pH meter can be seen in the
simplified diagram below.

The potential difference between the reference electrode and the glasselectrode is
amplified in the mV amplifier before the A/D converter feeds the signal to the microprocessor
for result calculation. To attain reliable and consistent results, the amplifier and other
circuitsmust have a small temperature coefficient, i.e. the influence of temperature variations
must be under control.
Normally, the result is displayed in numeric form although a few pH meterswith pointers
are still available. The term analog or digital pH meter is often used to distinguish between these
two forms of display. However, it is also used to differentiate between control convert circuitry
in analog or digital form.
In an analog pH meter, the adjustment of zero pH and sensitivity is carriedout using
adjustable resistances (dials) and the amplification factor is under direct manual control. The
signal is then sent through an A/D converter. The output is a digital signal for the numeric
display.
In a digital pH meter, the amplifier works under the same conditions all the time and is
directly connected to an A/D converter. The converter's output is then manipulated by digital
circuitry (microprocessor-based) and the calculated pH is then displayed. Use of a temperature
sensor provides both temperature correction and a temperature display.
To Measure pH of solution i.e. Measurement of pH:

A pH measurement system consists of a pH probe, reference probe, temperature sensor,

pH meter and the sample to be measured. In most cases the three probes are combined in one
electrode. When the pH probe is in contact with a solution a potential forms between the pH
probe and the reference probe. The meter measures the potential and converts it, using the
calibration curve parameters, into a pH value.
A typical combination glass electrode is represented as

In
order to measure the pH of a sample first the standardization of pH meter is required to be done
before analyzing the sample.
a) Standardization of pH meter:
A two point standardization method is used to standardize the pH meter, it
involves immersing the pH assembly i.e. glass electrode into a standard reference pH
buffer (pH =4.0) and recording the reading, if the meter reading is more or less than the
expected value (4.0) then it is adjusted to pH 4.0 using a crew nob.
Standardization at only one pH value does not assure the validity of reading at
other pH values considerably. Hence a second standard reference buffer pH = 9.2 is used.
The pH meter reading is recorded using this second buffer solution and the reading is
adjusted to pH 9.2 using a crew nob.
During both the steps, the glass electrode is rinsed with distilled water.Immerse
the glass electrode previously in water for several hours. Start the measurement more
than 5 minutes after switching on. Rinse well thedetecting unit with water, and blot the
water gently with a piece of filter paper very time.

b) To measure pH of solution:
Wash well the detecting unit with water, and blot the water gently with a piece of
filter paper. Place glass electrode in solution you wish to measure pH. Be sure that it is
stirring slowly during measure and pH adjustment and take readings.
c) Precautions:
When analysis is complete, put pH meter in stand-by mode. Remove electrode
from solution and rinse thoroughly with water. Blot dry and put back in yellow pH
storage buffer. Place parafilm over the hole and around the bottle to minimize
evaporation.
Glass Electrode:
Construction: It consists of a glass bulb membrane, which gives it its name and an electrically
insulating tubular body, which separates an internal solution and a silver/silver chloride electrode
from the studied solution. The Ag/AgCl electrode is connected to a lead cable terminated with
some connector that can hook up to a special voltmeter, the pH meter. It is represented as
Ag/AgCl | HCl | glass || probed solution | reference electrode

Working: The pH meter measures the potential difference and its changes across the glass
membrane. The potential difference must be obtained between two points; one is the electrode
contacting the internal solution. A second point is obtained by connecting to a reference
electrode, immersed in the studied solution.
The potential difference relevant to pH measurement builds up across the outside
glass/solution interface.
Eglass wall/solution = Eo - RT/2.303F log a(H3O+)
Where R is the molar gas constant 8.314 J mol-1 K-1, T is the temperature in kelvins, F is the
Faraday constant 96,485.3 C
 Eglass = E0 – 0.59 x log[H+]

Reference Electrode:
The reference electrode is a silver wire coated with silver chloride in contact with
a defined electrolyte solution, see above figure. In many reference electrodes a gel is used
instead of a liquid as the internal filling. These gels also contain KCl to maintain the
reference potential and add sufficient conductivity. As described in 2.2 the reference
potential is constant as long as the internal electrolyte is constant. The reference potential
also varies slightly with temperature.
The tube containing all the reference elements and solutions/gel is in contact with
the sample to measure through a junction (diaphragm). It is essential to maintain a free
flow of ions through the junction. Otherwise the reference electrode will not respond
properly to pH changes in the sample.
Eref = E0 – 0.59 x log[Cl-]

Combination Electrode:

Construction: The fig. shows the internal components of the pH electrode. The heart of the
electrode is a thin bulb of pH-sensitive glass, which is blown onto the end of a length of glass
tubing. The pH-sensitive glass (glass membrane) is sealed to the electrode and contains a
solution of potassium chloride at pH 7. A silver wire plated with silver chloride contacts the
solution. The Ag/AgCl combination in contact with the filling solution sets an internal reference
potential. This potential depends on the chloride concentration in the filling solution and as long
as this electrolyte concentration is maintained, the electrode potential is constant.
Working: The outside surface of the glass membrane is in contact with sample being
measured and the inside surface contacts the filling solution. A complex mechanism at each glass
liquid interface defines the potential the pH glass electrode, while the inner pH glass/ filling
solution potential is constant, the outside potential varies based on the H+ ions concentration in
sample. This equilibrium depends also on temperature.
 3. Conductometry

Conductometry is general method, where two electrode systems are used for simultaneous
measurement of all electroactive compounds in a solution. It is a measurement of electrolytic
conductivity to monitor a progress of chemical reaction.

Conductometric Titration:
Conductance is a property of solutions of electrolytes. It is the measure of the number of ions
present in the solution, as well as the current carrying capacity of the ions. When the solution
contains one single electrolyte, the measured conductance of the solution can be related to
concentration of the electrolyte.
Conductometric titration is a type of titration in which the electrolytic conductivity of the
reaction mixture is continuously monitored as one reactant is added. The equivalence point is the
point at which the conductivity undergoes a sudden change. Marked increase or decrease in
conductance is associated with the changing concentrations of the two most highly conducting
ions, viz. the hydrogen and hydroxyl ions. The method can be used for titrating coloured
solutions or homogeneous suspension e.g.: wood pulp suspension, which cannot be used with
normal indicators.
Principle:
When solution of one electrolyte is added to another electrolyte without appreciable
volume change, the conductance of the solution will alter, if an ionic reaction occurs. If no ionic
reaction takes place then the conductance of the solution will simply increase. On the other hand,
if an ionic reaction occurs, the ion added may replace another ion and hence bring about change
in the conductance.
Let A+B- be the ions of titrand and C+D- be ions of the titrant, the ionic reaction in the
titration is combination of A+ and D-, AD formed may be insoluble or weakly ionized.
A+B- + C+D-  AD + C+B-
Thus as the titration proceeds, A+ are replaced by C+. The conductance of the solution
increases or decreases depending on whether conductance of C+ is greater than or less than that
of A+. After equivalence point the ionic reaction does not occur and hence, the conductance of
the solution will raise due the excess addition of titrant C+D-.
The principle of conductometric titration is changes in the conductance of the solution
due to difference in the ionic conductance or due to production of more number of ions in the
solution. Both factors permit, location of the equivalence point by conductance measurements.
Procedure:
A definite volume of the solution to be estimated is pipetted out in a beaker. A dip type
conductivity cell is place in a beaker. Addition of distilled water may be necessary if the cell
does not dip completely in the solution. The cell is connected to a conductometer and the
conductance of the solution is measured. The titrant is filled in the burrete. The titrant is added in
the small portions, generally 0.5 mL at a time. The solution is stirred after each addition. The
solution is allowed to stand for a minute or two after stirring before conductance is measured.
Addition of titrant is continued till about seven to eight readings beyond the equivalence point
are obtained. The plot of conductance against volume of the titrant added is used to locate the
equivalence point.
Titration Curves:
Some Typical Conductometric Titration Curves are:
1) Strong Acid with a Strong Base, [HCl Vs NaOH]
Before NaOH is added, the conductance is high due to the presence of highly mobile
hydrogen ions. When the base is added, the conductance falls due to the replacement of
hydrogen ions by the added cation as H+ ions react with OH- ions to form undissociated
water. This decrease in the conductance continues till the equivalence point. At the
equivalence point, the solution contains only NaCl. After the equivalence point, the
conductance increases due to the large conductivity of OH- ions

2) Weak Acid with a Strong Base, [CH3COOH Vs NaOH] Initially the conductance is
low due to the feeble ionization of acetic acid. On the addition of base, there is
decrease in conductance not only due to the replacement of H+ by Na+ but also
suppresses the dissociation of acetic acid due to common ion acetate. But very soon,
the conductance increases on adding NaOH as NaOH neutralizes the un-dissociated
CH3COOH to CH3COONa which is the strong electrolyte. This increase in conductance
continues raise up to the equivalence point. The graph near the equivalence point is
curved due the hydrolysis of salt CH3COONa. Beyond the equivalence point,
conductance increases more rapidly with the addition of NaOH due to the highly
conducting OH-ions.
3) Strong Acid with a Weak Base, e.g. sulphuric acid with dilute ammonia: Initially
the conductance is high and then it decreases due to the replacement of H+. But after
the endpoint has been reached the graph becomes almost horizontal, since the excess
aqueous ammonia is not appreciably ionised in the presence of ammonium sulphate.

4) Weak Acid with a Weak Base: The nature of curve before the equivalence point is
similar to the curve obtained by titrating weak acid against strong base. After the
equivalence point, conductance virtually remains same as the weak base which is being
added is feebly ionized and, therefore, is not much conducting.
 5) Mixture of a Strong Acid and a Weak Acid vs. a Strong Base or a Weak Base: In
this curve there are two break points. The first break point corresponds to the
neutralization of strong acid. When the strong acid has been completely neutralized
only then the weak acid starts neutralizing. The second break point corresponds to the
neutralization of weak acid and after that the conductance increases due to the excess
of OH−ions in case of a strong base as the titrant. However, when the titrant is a weak
base, it remains almost constant after the end point similar to Fig.

Conductometric titration of a mixture of a strong acid (HCl) and a weak

acid (CH3COOH) vs. a strong base (NaOH) or a weak base (NH4OH)
Advantages of Conductometric titrations:
Conductometric titration are found to possess several advantages over normal titrimetry
namely,
1. Coloured solutions can be titrated.
2. The method works equally well for dilute solutions also, as it is based on the changes in
the conductance, rather than the absolute value of the conductance.
3. In conductometric titrations, it is not necessary to make observations around the
equivalence point, with small increments of the titrant added. The observations, far away
from the equivalence point, on either side are of importance.
4. An extremely weak acid or a weak base can be titrated conductometrically, which may
not be possible in normal titrimetry.
5. A mixture of weak and strong acids, can also be titrated with relative ease. Thus, making
the simultaneous determination possible.

Limitations:
1. In dilute solutions, obtuse curves are obtained. With obtuse curves it is difficult to locate
the equivalence point accurately.
2. The overall accuracy of the conductometric titrations is limited as the technique does not
permit addition of small increments of the titrant.
 4. IR spectroscopy

In contrast to ultraviolet spectroscopy IR spectrum provides a rich array of absorption

bands which can provide accurate structural information about a molecule. It provides the
methods for studying materials in all three physical states i.e. solid, liquid and gas. Analytically
useful IR spectrum covers the following range of electromagnetic spectrum.

Near IR 15000 cm-1 to 3000 cm-1

Mid IR 4000 cm-1 to 400 cm-1
Far IR 200 cm-1 to 10 cm-1

Most used 4000 cm-1 to 670 cm-1

Infrared radiation is largely thermal energy. It induces stronger molecular vibrations in

covalent bonds, which can be viewed as springs holding together two masses, or atoms. Specific
bonds respond to (absorb) specific frequencies.

VIBRATIONAL MODES:
The information contained in IR spectrum originates from molecular vibrations. These
are either fundamental vibrational modes that are associated with the vibrations of specific
functional group, or molecule, vibrational overtones or summational modes of fundamental
vibrations.
A molecule resembles a system of balls of varying masses corresponding to atoms of a
molecule and spring of varying lengths corresponding to various chemical bonds. There are two
fundamental vibrational modes.
1. Stretching: in which the distance between the two atoms increases or decreases but
the atoms remain in the same bond axis.
2. Bending: in which the position of the atom changes relative to the bond axis.
Covalent bonds can vibrate in several modes, including stretching, rocking, and
scissoring.
 The various stretching and bending vibrations occurs at certain frequencies. When an IR
radiation of same frequency is incident on the molecule, the energy is absorbed and the
amplitude of that vibration increases correspondingly. When the molecule returns to ground state
the absorbed energy released as heat.
A nonlinear molecule containing n atoms has 3n-6 possible vibrational modes through
which IR radiation may be absorbed. For example methane has 9 and benzene has 30 possible
fundamental absorption bands respectively. In order that a particular vibration results in an
absorption band, the vibration must cause change in the dipole moment of the molecule.

Which substances give a signal in IR spectrum?

Ans: The molecules that contain polar bonds i.e. molecules composed of
atoms of different elements, organic compounds and inorganic compounds (H2O,
CO2, NO2, HCl, salts...) can give a signal in IR spectrum. Whereas pure chemical elements in
molecular or crystal state e.g. Ar, O2, O3, N2, Cl2, S8, silicon, graphite,
Diamond etc cannot give a signal in IR spectrum.

Basic Principle:
When a sample is placed in a beam of infrared radiation, the sample will absorb radiation
at frequencies corresponding to molecular vibrational frequencies, but will transmit all other
frequencies. The frequencies of radiation absorbed are measured by an infrared spectrometer,
and the resulting plot of absorbed energy vs. frequency is called infrared spectrum of the
material. Identification of a substance is possible because different materials have different
vibrations and yield different infrared spectra. Furthermore, from the frequencies of the
absorption it is possible to determine whether various chemical groups are present or absent in a
chemical structure.
Instrumentation of FTIR:
The basic components of an FTIR are shown schematically in fig.

1. The Source:- Infrared energy is emitted from a glowing black body source. This beam
passes through an aperture which controls the amount of energy presented to the sample (and,
ultimately, to the detector).
2. The Interferometer:- The beam enters the interferometer where the “spectral
encoding” takes place. The resulting interferogram signal then exits the interferometer.
3. The Sample:- The gaseous sample can be directly analysed. Liquid can also be used
directly but in diluted form in NaCl plates. Solid compound can be mixed with KBr and formed a
pallet and used.
The beam enters the sample compartment where it is transmitted through or reflected off
of the surface of the sample, depending on the type of analysis being accomplished. This is
where specific frequencies of energy, which are uniquely characteristic of the sample, are
absorbed.
4. The Detector:- The beam finally passes to the detector for final measurement. The
detectors used are specially designed to measure the special interferogram signal.
5. The Computer:- The measured signal is digitized and sent to the computer where the
Fourier transformation takes place. The final infrared spectrum is then presented to the user for
interpretation and any further manipulation.

Working:
The infrared source emits a broad band of different wavelength of infrared radiation. The
IR source used is a SiC ceramic at a temperature of 1550 K. The IR radiation goes through an
interferometer that modulates the infrared radiation. The interferometer performs an optical
inverse Fourier transform on entering IR radiation. The modulated IR beam passes through the
gas sample where it is absorbed to various extents at different wavelengths by the various
molecules present. Finally, the intensity of the IR beam is detected by a detector, which is a
liquid nitrogen cooled MCT (Mercury−Cadmium−Telluride) detector. The detected signal is
digitised and Fourier transformed by the computer to get the IR spectrum of the sample gas.

AN IR SPECTRUM IN ABSORPTION MODE:

The IR spectrum is basically a plot of transmitted (or absorbed) frequencies
vs intensity of the transmission (or absorption). Frequencies appear in the x-axis in units of
inverse centimeters (wavenumbers), and intensities are plotted on the y-axis in percentage units.
AN IR SPECTRUM IN TRANSMISSION MODE:

The graph above shows a spectrum in transmission mode. This is the most commonly
used representation and the one found in most chemistry and spectroscopy books.

CLASSIFICATION OF IR BANDS
IR bands can be classified as strong (s), medium (m), or weak (w), depending on their
relative intensities in the infrared spectrum. A strong band covers most of the y-axis. A medium
band falls to about half of the y-axis, and a weak band falls to about one third or less of the y-
axis.
 Infrared band shapes come in various forms. Two of the most common are narrow and
broad. Narrow bands are thin and pointed, like a dagger. Broad bands are wide and smoother.
A typical example of a broad band is that displayed by O-H bonds, such
as those found in alcohols and carboxylic acids, as shown below.

INFORMATION OBTAINED FROM IR SPECTRA

• IR is most useful in providing information about the presence or absence of specific functional
groups.
• IR can provide a molecular fingerprint that can be used when comparing samples. If two pure
samples display the same IR spectrum it can be argued that they are the same compound.
• IR does not provide detailed information or proof of molecular formula or structure. It provides
information on molecular fragments, specifically functional groups.
• Therefore it is very limited in scope, and must be used in conjunction with other techniques to
provide a more complete picture of the molecular structure.
THE FINGERPRINT REGION
Although the entire IR spectrum can be used as a fingerprint for the purposes of
comparing molecules, the 600 - 1400 cm-1 range is called the fingerprint region.
This is normally a complex area showing many bands, frequently overlapping each other.
It is much more difficult to pick out individual bonds in this region than it is in the
"cleaner" region at higher wavenumbers. The importance of the fingerprint region is that each
different compound produces a different pattern of troughs in this part of the spectrum.

FUNCTIONAL GROUPS AND IR TABLES:

Characteristic IR Absorption Frequencies of Organic Functional Groups
Functional Type of Characteristic
Intensity
Group Vibration Absorptions (cm-1)
Alcohol
(stretch, H-
O-H 3200-3600 strong, broad
bonded)
O-H (stretch, free) 3500-3700 strong, sharp
C-O (stretch) 1050-1150 strong
Alkane
C-H stretch 2850-3000 strong
-C-H bending 1350-1480 variable
Alkene
=C-H stretch 3010-3100 medium
=C-H bending 675-1000 strong
C=C stretch 1620-1680 variable
Alkyl Halide
C-F stretch 1000-1400 strong
C-Cl stretch 600-800 strong
C-Br stretch 500-600 strong
C-I stretch 500 strong
Alkyne
C-H stretch 3300 strong,sharp
variable, not present in symmetrical
stretch 2100-2260
alkynes
Amine
medium (primary amines have two
N-H stretch 3300-3500 bands; secondary have one band, often
very weak)
C-N stretch 1080-1360 medium-weak
N-H bending 1600 medium
Aromatic
C-H stretch 3000-3100 medium
C=C stretch 1400-1600 medium-weak, multiple bands
Analysis of C-H out-of-plane bending can often distinguish substitution patterns
Carbonyl Detailed Information on Carbonyl IR
C=O stretch 1670-1820 strong
(conjugation moves absorptions to lower wave numbers)
Ether
1000-1300 (1070-
C-O stretch strong
1150)
Nitrile
CN stretch 2210-2260 medium
Nitro
1515-1560 & 1345-
N-O stretch strong, two bands
1385

IR Absorption Frequencies of Functional Groups Containing a Carbonyl (C=O)

Functional Type of Characteristic Absorptions
Intensity
Group Vibration (cm-1)
Carbonyl
C=O stretch 1670-1820 strong
(conjugation moves absorptions to lower wave numbers)
Acid
C=O stretch 1700-1725 strong
O-H stretch 2500-3300 strong, very broad
C-O stretch 1210-1320 strong
Aldehyde
C=O stretch 1740-1720 strong
=C-H stretch 2820-2850 & 2720-2750 medium, two peaks
Amide
C=O stretch 1640-1690 strong
unsubstituted have two
N-H stretch 3100-3500
bands
N-H bending 1550-1640
Anhydride
C=O stretch 1800-1830 & 1740-1775 two bands
Ester
C=O stretch 1735-1750 strong
C-O stretch 1000-1300 two bands or more
Ketone
acyclic stretch 1705-1725 strong
3-membered - 1850
4-membered - 1780
cyclic Stretch 5-membered - 1745 strong
6-membered - 1715
7-membered - 1705
, -unsaturated Stretch 1665-1685 strong
aryl ketone Stretch 1680-1700 strong

How to analyse the IR spectrum?

 When analysing the IR spectrum of an unknown molecule, first efforts on determining
the presence or absence of a few major functional groups. The C=O, O-H, N-H, C-O, C=C,
, -CN, and NO2 peaks. These peaks are most conspicuous and give immediate
structural information if they are present. Do not try to make a detailed analysis of the C-H
absorption near 3000cm-1, almost all the compounds have these absorptions.
Follow the steps
1. Is carbonyl group is present? The –C=O group give rise to a strong absorption in the
region 1820-1660 cm-1. The peak is often stongest in the spectrum and of medium width.
2. If C=O is present, then check the following types
3.
Acids Is O-H also present? Broad absorption near 3400-2400cm-1 usually
overlaps with C-H
Amides Is N-H also present? Medium absorption near 3400cm-1, sometimes
double peaks with same size.
Esters Is C-O present? Strong intensity absorption near 1300-1000cm-1
Anhydrides Two C=O absorption near 1810 and 1760cm-1
Aldehydes Is aldehyde C-H present? Two weak absorption near 2850 and 2750
cm-1
Ketones The preceding five choices have been eliminated
4. If C=O absent, then check the following options
Alcohols, Check for O-H, broad absorption near 3400-3300cm-1, confirm this by
Phenols finding C-O near 1300-1000cm-1
Amines Check for N-H, Medium absorptions near 3400cm-1
Ethers Check for C-O near 1300-1000cm-1 and absence of O-H near 3400cm-1
5. Double bonds and/or aromatic rings
1. C=C is weak absorption near 1650cm-1
2. Medium strong absorption in the region 1600-1450cm-1, these often imply on
aromatic ring
3. Confirm the double bond or aromatic ring by consulting the C-H region; aromatic
and vinyl C-H occurs to left of 3000cm-1, aliphatic C-H occurs to right of this
value.
4. Triple bonds
1) C N is medium sharp absorption near 2250cm-1

2) C Cis e weak, sharp absorption near 2150cm-1

3) Check also for acetylenic C-H near 3300cm-1

5. Nitro groups
 Two strong absorption at 1600-1530cm-1 and 1390-1300cm-1
6. Hydrocarbons
None of the preceding found.
Major absorptions are in C-H region near 3000cm-1
Very simple structure the only another absorption appear near 1460 and 1375cm-1

IR spectrum of n-Hexane

IR spectrum of 1-Hexene
IR spectrum of 1-Octyne and 4-Octyne

IR spectrum of Butyronitrile

IR spectrum of 1-Butnol

IR spectrum of 1-Butanal and 2-Heptanone

IR spectrum of n-Hexanoic acid
IR spectrum of 1-Butanamide

IR spectrum of 3-Bromoaniline