Roll No.

Name
Batch

CHEMISTRY LABORATORY INSTRUCTIONS

AND OBSERVATION BOOK

FOR

I SEMESTER - B. E. / B. TECH.

(FOR PRIVATE CIRCULATION ONLY)

DEPARTMENT OF CHEMISTRY
ANNA UNIVERSITY
CHENNAI 600 025

1
 CONTENTS
(9 experiments to be conducted)

Page
S. No. Contents
number

Estimation of hydrochloric acid using sodium carbonate

1. 9
as primary standard

Determination of type and estimation of amount of

2. 13
alkalinity in the given water sample

Estimation of total, temporary and permanent hardness

3. 19
of the given water sample by EDTA titration

Estimation of dissolved oxygen in water by Winkler's

4. 25
method

Estimation of amount of copper in the given water sample

5. 29
by iodometry

Determination of molecular weight of a polymer by

6. 33
viscometry

Estimation of iron in the given sample by spectrophotometric

7. 38
method

Estimation of the strength of acids in a mixture by

8. 44
conductometric titration

Estimation of ferrous ion in the given solution by

9. 50
potentiometric titration

Estimation of sodium ion in water sample by flame

10. 56
photometry

Determination of the strength of the given HCI by pH

11. 60
metric titration

2
 SCHEDULE OF EXPERIMENTS
Week 1 Estimation of HCI using sodium carbonate as primary standard
Week 2 Determination of alkalinity
Determination of total, temporary and permanent hardness of the given
Week 3
water sample by EDTA method
Week 4 i) DO estimation
Week 5 i) Copper content estimation
i) Viscometric determination of molecular weight of PVA
Week 6
ii) Determination of Iron by visible spectrophotometry
i) Potentiometry
Week 7
ii) Conductometric titration
Week 8 Revision and Record Book submission
Week 9 End semester Laboratory examination

3
 INDEX

SIGNATURE
S. NAME OF THE PAGE OF FACULTY-
DATE
No. EXPERIMENT No. IN-CHARGE
WITH DATE

4
 GENERAL INFORMATION ON PRACTICAL CHEMISTRY

 Chemical analysis:

o Qualitative analysis : To find what is present in the given sample.

o Quantitative analysis: To estimate the amount of desired species present in the

given sample.

 Quantitative analysis can be performed by:

o Wet chemical analysis and

o Instrumental analysis.

o Wet chemical analysis: a) Titrimetry (Volumetry) and b) Gravimetry.

o Instrumental analysis:

a) Thermal analysis - DSC, TGA and DTA.

b) Electro analytical methods - Electrogravimetry, Potentiometry, Coulometry

and Voltammetry.

c) Spectral methods - IR spectroscopy, UV-Visible spectrophotometry, Flame

photometry and Atomic absorption spectroscopy.

d) Chromatography - Thin layer chromatography, Gas chromatography ar

Liquid chromatography.

 Titrimetry: Classified as: Acid-base (neutralisation) titrations, ii) Complexometric

titrations, iii) Precipitation titrations and iv) Redox titrations.

 Molecular weight of organic and inorganic compounds = sum of atomic weights of

the elements.

Example: Molecular weight of NaOH = 23 + 16 + 1 = 40 g/mol

 Equivalent weight of a compound = Molecular weight / Acidity or basicity of the

same

Example:

a) For NaOH, Acidity= 1

Equivalent weight of NaOH = Molecular weight = 40 g/ eq.

b) For H2SO4 (2H+ + SO 42-), basicity is 2

Molecular weight of H2SO4 = 98.07 g/mol
Equivalent weight of H2SO4 = (Molecular weight /2) = 98.07/2 = 49.03 g/eq

5
 Concentration of solutions can be expressed in terms of:

Weight of solute per litre of solution

i) Molarity (M) =
Molecular weight of the solute

Weight of solute per litre of solution

ii) Normality (N) =
Equivalent weight of the solute

Weight of solute per kilogram of solvent

iii) Molality (m) =
Molecular weight of the solute

iv) Percentage solutions = solute in gram per 100 mL of solvent.

e.g.: A 10% NaCl solution refers to 10 g of NaC1 in 100 mL of water.

ppm (or) mg/ L - parts per million which is the same as mg per litre.

 Indicators: Chemical compounds capable of indicating the end points in titrations

through colour change.

Examples:

 Acid-base indicators - phenolphthalein, methyl orange, methyl red, congo red

 Adsorption indicators - fluorescein, tartrazine

 Oxidation - reduction indicators - starch, diphenylamine, methylene blue,

1, 10 - Phenanthroline

 Metal ion indicators - Eriochrome Black -T, xylenol orange, murexide, calcichrome

 Self indicating species - potassium permanganate

6
DATA AND CALCULATION

I. PREPARATION OF STANDARD SODIUM CARBONATE SOLUTION

Weight of anhydrous sodium carbonate = g / 100 mL

Equivalent weight of anhydrous sodium carbonate = 53 g/eq

Weight / L
Strength of sodium carbonate = ----------------------
Equivalent weight

= x10
----------------------
53

= N

2. ESTIMATION OF THE GIVEN HYDROCHLORIC ACID SAMPLE

Standard Na2CO3 solution Vs the given HCl

Volume of standard Burette reading (mL) Volume Concordant

S. No. Na2CO3 solution of HCl of value Indicator
(mL) Initial Final (mL) (mL)

Methyl
orange

7
Expt. No.: Date:

ESTIMATION OF HYDROCHLORIC ACID USING SODIUM CARBONATE AS

PRIMARY STANDARD
AIM

To prepare a standard solution of sodium carbonate of accurately 0.1 N strength and estimate
the amount of hydrochloric acid present in the given sample using standard sodium carbonate
solution.

PRINCIPLE

A standard solution is the one whose strength is accurately known. The concentration of
solutions used in chemical analysis is generally expressed in normality (N) or molarity (M).

Primary standards are prepared from solutes which are: i) available in the pure form, ii) stable
towards environment, iii) relatively inexpensive and iv) readily soluble to give a stable solution
in which the gram equivalent of the substance is large so that the strength can be determined with
utmost precision.

Examples: Anhydrous sodium carbonate, potassium dichromate, Mohr's salt, oxalic acid.

Secondary standards are prepared from solutes which do not satisfy the above criteria but
give rise to stable solutions whose strength can be determined accurately only by standardisation
using a primary standard.

Examples: NaOH, KOH, HC1, Sodium thiosulphate.

In preparing the primary standard, a calculated quantity of the solute is exactly weighed,
dissolved in the solvent and made up to a known volume in a standard measuring flask (SMF). For
the standardisation of the secondary standard, a known volume of the primary standard is titrated
against the secondary standard. From the strength of the secondary standard, the amount of solute
in g/L can be estimated as:

Amount in g/L = Normality x Equivalent weight

PROCEDURE

I. Preparation of standard sodium carbonate solution

About 0.53 g of anhydrous sodium carbonate is exactly weighed and transferred into a
100 mL SMF through a funnel. Distilled water is poured through the funnel in small amounts
to wash down the salt into the flask. The funnel is washed and removed after ensuring complete
transfer of the salt into the flask. The flask is gently swirled to dissolve the salt and the volume is
made up to the mark in the SMF by adding distilled water. The SMF is then stoppered and shaken
to ensure uniform concentration of the solution.

8
Volume of Na2CO3 solution (V1) = 20 mL

Strength of Na2CO3 solution (N1) = N

Volume of the given HC1 solution (V2) = mL
Strength of the given HCl solution (N2) = (V1 x N1) / V2

= N

Amount of HC1 = N2 x Equivalent weight of HC1

Equivalent weight of HC1 = 36.5 g/eq.

= g/L

9
2. Estimation of the given hydrochloric acid sample

20 mL of the standard sodium carbonate solution is pipetted out into a clean

conical flask and 2-3 drops of methyl orange indicator is added to it. The solution is
yellow in colour. The content of the conical flask is titrated against the given HC1
taken in the burette. The end point is the change in colour from yellow to reddish
orange. The titration is repeated for concordant values.

RESULT:

1. Strength of the given HC1 = N.

2. The amount of HC1 present in the given sample = g/L.

10
DATA AND CALCULATION

TITRATION 1: STANDARDISATION OF HC1

Standard Na2CO3 solution Vs HCl
S. No. Volume of Burette reading Volume of Concordant Indicator
standard Na2CO3 (mL) HCl required value (mL)
solution (mL) Initial Final (mL)

Methyl
orange

Volume of Na2CO3 solution (V1) = mL

Strength of Na2CO3 solution (N1) = N
Volume of HCl solution (V2) = mL
Strength HC1 solution (N2) = (V1 x N1) / V2

=
= N

TITRATION 2: ESTIMATION OF ALKALINITY OF THE GIVEN WATER SAMPLE

Titration of the given water sample Vs standardised HCl
S. Volume of Burette reading (mL) Volume of Concordant
No. the given Initial Final HC1 consumed value
water At At (mL)
sample phenolphthalein methyl orange
(mL) P M P M
end point end point (mL) (mL)
P M
Expt. No.: Date:

DETERMINATION OF TYPE AND ESTIMATION OF AMOUNT OF ALKALINITY

IN THE GIVEN WATER SAMPLE

AIM
To determine the type and estimate the amount of alkalinity present in the given water sample.
PRINCIPLE
Alkalinity in water is due to the presence of soluble hydroxides, carbonates and / or
bicarbonates. The type and amounts of alkalinity present in the water sample can be determined
by titrating the sample against a strong acid solution of known strength using phenolphthalein and
methyl orange indicators in succession.
The neutralisation of different types of alkalinity by acid can be represented as follows:

1. OH-
2. CO32-
3. HCO3-
+
+
+
H+
H+
H+
H2O
HCO3-
H2CO3
}
Indicated by
phenolphthalein end
point
H2O + CO2 
} Indicated by
methyl orange
end point

The type of alkalinity can be found out from the volume of phenolphthalein end point (P) and
methyl orange end point (M) as follows:
i) If P = M, water sample contains only OH- alkalinity, corresponding to neutralisation
reaction 1.
ii) If P = ½ M (or) 2P = M, water sample contains only CO3 2- alkalinity.
The phenolphthalein end point corresponds to half-neutralisation of CO 3 2- and further
neutralisation of the generated HCO3- (Reaction 2) by acid is determined using methyl orange
indicator. Thus, methyl orange end point indicates the complete neutralisation of CO3 2- ions.
iii) If P = 0 and M > 0, water sample contains only HCO 3- alkalinity.
iv) If P > ½ M (or) 2P > M, water sample contains both OH- and CO32- alkalinity.
P = OH- + ½ CO 32-
M = OH- + CO 32-

ç OH- alkalinity = 2P – M and

CO32- alkalinity = 2 (M – P)
 P= mL and M= mL

If 2P > M, then, the water sample contains OH - and CO3 2- alkalinity

The amount of individual and total alkalinity can be calculated as follows:

1) Alkalinity due to OH- :

Volume of acid corresponding to OH- alkalinity (V1 ) = 2P - M = = mL

Strength of acid (N1) = N

Volume of the given water sample (V2) = 20 mL

Strength of CO 2- alkalinity in the given water sample (N ) = --------------- = N
3 2
20

Alkalinity due to OH- = N 2x 50 x 1000 mg/L of CaCO eq.

3

= mg/L of CaCO3 eq.

2. Alkalinity due to CO
3
2-
:

Volume of acid corresponding to CO3 2- alkalinity (V1) = 2 (M - P) = = mL

Strength of acid (NI) = N

Volume of the given water sample (V2) = 20 mL

Strength of CO 2- alkalinity in the given water sample (N ) = -------------- = N
3 2
20

Alkalinity due to CO3 2- = N2 x 50 x 1000 mg/L of CaCO3 eq.

= mg/L of CaCO3 eq.

Total alkalinity of the given water sample = OH- alkalinity + CO32- alkalinity
=

= mg/L, of CaCO3 eq.

v) If P <½ M (or) 2P < M, water sample contains CO 2-3 and HCO -3alkalinity.
P = ½ CO 2-3
M = CO 32- + HCO -3
ç CO32- alkalinity = 2P and
HCO3- alkalinity = M — 2P
Hydroxyl and bicarbonate ions will not exist together since they react to form CO 2-3 .
OH - + HCO 3-  H O
2
+ CO 2-
3

ALKALINITY CONDITIONS
[Using Phenolphthalein [P] and Methyl Orange [M] values]

S. No. Result of Titration Alkalinity causing ions

[P] and [M] CO32- HCO3-
OH-
1. [P] = 0 Nil Nil [M]
2. [P] = [M] [P] or [M] Nil Nil
3. [P] = 1/2 [M] Nil 2 [P] or [M] Nil
4. [P] > 1/2 [M] 2[P] - [M] 2 [M] — 2 [P] Nil
5. [P] < 1/2 [M] Nil 2 [P] [M] — 2 [P]

PROCEDURE
Titration 1: Standardisation of hydrochloric acid
20 mL of the standard sodium carbonate solution is pipetted out into a 250 mL clean conical
flask and 2-3 drops of methyl orange indicator is added to it. The colour of the solution turns
yellow. The content of the conical flask is titrated against HC1 solution taken in the burette. The
end point is the change in colour from yellow to reddish orange. The titration is repeated for
concordant values.
Titration 2: Estimation of alkalinity of the given water sample
20 mL of the given water sample is pipetted out into a clean conical flask. 2-3 drops of
phenolphthalein indicator is added to it and the content of the flask is titrated against the
standardised HCI solution. The end point is the disappearance of pink colour and it is noted
as phenolphthalein end point (P). About 4-5 drops of methyl orange indicator are added to the
same solution and without tilling the burette the titration is continued till the colour changes
from yellow to reddish orange. The final titre value corresponds to the complete neutralisation
of alkalinity and it is noted as the methyl orange end point (M). The titration is repeated for
concordant values.
If 2P < M, then, the water sample contains CO3 2- and HCO3 - alkalinity

The amount of individual and total alkalinity can be calculated as follows:

1) Alkalinity due to CO 32-

Volume of acid corresponding to CO3 2- alkalinity (V1 ) = 2P = = mL

Strength of acid (N1) = N

Volume of the given water sample (V2) = 20 mL

Strength of CO3 2- alkalinity in the given water sample (N2) = --------------- = N

20

Alkalinity due to CO32- = N 2 x 50 x 1000 mg/L of CaCO3 eq.

= mg/L of CaCO3 eq.

2. Alkalinity due to HCO

3
-

Volume of acid corresponding to HCO3 - alkalinity (V1) = M - 2P = = mL

Strength of’ acid (NI) = N

Volume of the given water sample (V2) = 20 mL

Strength of HCO - alkalinity in the given water sample (N ) = -------------- = N
3 2
20

Alkalinity due to HCO3- = N2 x 50 x 1000 mg/L of CaCO3 eq.

= mg/L of CaCO3 eq.

Total alkalinity of the given water sample = CO32- alkalinity + HCO3- alkalinity

= mg/L, of CaCO3 eq.

 The type of alkalinity is determined from the values of P and M and individual
alkalinity is expressed in terms of mg/L or CaCO3 equivalent.

RESULT

1. The types of alkalinity present in the given water sample are :

2. Amount of alkalinity = mg/L of CaCO3 eq.

3. Amount of alkalinity = mg/L of CaCO3 eq.

4. Total alkalinity of water sample = mg/L of CaCO3 eq.

6
DATA AND CALCULATION

TITRATION 1: STANDARDISATION OF EDTA

Standard hard water Vs EDTA

S. No. Volume of standard Burette reading Volume Concordant Indicator

hard water (mL) (mL) of EDTA value
Initial Final (mL) (mL)

Eriochrome
Black -T

Volume of standard hard water (V1) = mL

Strength of standard hard water (M1) = 0.01 M

Volume of EDTA solution (V2) = mL

ç Strength of EDTA solution (M2) = (V1 x M1) / V2

= M
Expt. No.: Date:

ESTIMATION OF TOTAL, TEMPORARY AND PERMANENT HARDNESS OF THE

GIVEN WATER SAMPLE BY EDTA TITRATION

AIM

To estimate the total, temporary and permanent hardhess of the given water sample by EDTA
method.

PRINCIPLE

Hardness in water is due to the presence of dissolved bicarbonates, chlorides and sulphates
of calcium and magnesium. The amount of hardness in a sample of water can be estimated by
complexing the calcium and magnesium ions using disodium salt of ethylene diamine tetraacetic
acid (EDTA). EDTA is a hexadentate ligand capable of forming a soluble, stable complex with the
divalent cations.

Structure of EDTA

In the EDTA method for the estimation of hardness, Eriochrome Black -T [EBT, an azo dye
: l-(1-Hydroxy-2-naphthylazo)-6-nitro-2-naphthol-4-sulphonate] is used as an indicator. A pH of
9.5± 0.5 necessary for the complexation reaction is maintained by adding an ammoniacal buffer
consisting of a mixture of NH4C1 and NH4OH to the hard water sample.
When the indicator is added to the buffered hard water sample, EBT forms a wine red coloured,
less-stable, weak complex with few calcium and magnesium ions of hard water.

When this sample is titrated against EDTA of known strength, all the free un-complexed
calcium and magnesium ions are trapped to form a colourless, stable metal ion-EDTA complex.
 TITRATION 2: ESTIMATION OF TOTAL HARDNESS OF THE GIVEN WATER
SAMPLE
Hard water sample Vs. standardised EDTA
S. No. Volume of hard Burette reading Volume Concordant Indicator
water sample (mL) of EDTA value (mL)
(mL) Initial Final (mL)

Eriochrome
Black -T

Volume of EDTA solution (V1) = mL

Strength of EDTA solution (M1) = M
Volume of hard water sample (V2) = mL
.•. Strength of hard water sample (M2) = (V1 x M1) / V2
=
= M

Total hardness in terms of CaCO 3 eq. =

=
[Strength of
hard water sample x 100 x 1 000 mg/L
x 100 x 1000
Total hardness of the given water sample = mg/L of CaCO3 eq.
 Once the complexation of all the free Ca 2+ and Mg2+ ions is complete, further addition of
EDTA will result in the capture of the cations from the indicator complex. This leads to the release
of the indicator to the solution. Hence, the end point in this EDTA titration is the change in colour
from wine red (indicator complex) to steel blue (original colour of EBT).

pH = 9.5± 0.5
[Ca / Mg - EBT] + EDTA
2+ 2+
[Ca2+ / Mg2+ - EDTA] + EBT
Wine red Steel blue

Hardness of the water sample can be calculated from the titre value and expressed in terms of
mg/L of calcium carbonate equivalent.
TITRATION 3: ESTIMATION OF PERMANENT HARDNESS OF THE GIVEN WATER
SAMPLE
Boiled and filtered Hard water sample Vs standardised EDTA
S. No. Volume of boiled & Burette reading Volume Concordant Indicator
filtered hard water (mL) of EDTA value (mL)
sample (mL) Initial Final (mL)
Eriochrome
Black -T

Volume of EDTA solution (V1) = mL

Strength of EDTA solution (M1) = M

Volume of boiled and filtered

hard water sample (V2) = mL

Strength of boiled and filtered

hard water sample (M2) = (V1 x M1) / V2

= M

ç Permanent hardness in terms of CaCO 3 eq. = M2 x 100 x 1000 mg/L

= x 100 x 1000

Permanent hardness of the given water sample = mg/L of CaCO3 eq.

ç Temporary hardness = Total hardness — Permanent hardness

= mg/L of CaCO3 eq
PROCEDURE

Titration 1: Standardisation of EDTA

20 mL of the given standard hard water (0.01M) is pipetted out into a clean conical flask.
About 10 mL of NH4C1-NH4OH buffer solution and a pinch of Eriochrome Black -T indicator are
added to it. The content of the conical flask is then titrated against the EDTA solution taken in the
burette. The end point is the change in colour from wine red to steel blue. Titration is repeated for
concordant values.

Titration 2: Estimation of total hardness of the given water sample

20 mL of the given water sample is pipetted out into a clean conical flask. About 10 mL of
NH4C1-NH4OH buffer solution and a pinch of Eriochrome Black -T indicator are added to it. The
content of the conical flask is titrated against the EDTA solution taken in the burette. The end point
is the change in colour from wine red to steel blue. Titration is repeated for concordant values.

Titration 3: Estimation of permanent hardness of the given water sample

About 50 mL of the given hard water sample is taken in a 250 mL beaker and is boiled well,
until the volume reduces to about 25 mL. The water is then cooled, filtered into a 100 mL standard
flask and made up to the mark using distilled water. 20 mL of this water is pipetted out into a clean
conical flask. 10 mL of buffer solution and a pinch of Eriochrome Black -T indicator are added to
it. The solution is then titrated against EDTA taken in the burette. The end point is colour change
from wine red to steel blue.

RESULT

Permanent hardness of the given water sample = mg/L of CaCO3 eq.

Temporary hardness of the given water sample = mg/L of CaCO3 eq.

Total hardness of the given water sample = mg/L of CaCO3 eq.

7
DATA AND CALCULATION

TITRATION 1: STANDARDISATION OF SODIUM THIOSULPHATE

Standard potassium dichromate Vs sodium thiosulphate

S. Volume of standard Burette reading Volume of Concordant Indicator

No. potassium dichromate (mL) sodium thiosulphate value
(mL) Initial Final solution (mL) (mL)

Starch

Volume of standard potassium dichromate (V1) = mL

Strength of standard potassium dichromate (N1) = N

Volume of sodium thiosulphate solution (V2) = mL

:. Strength of sodium thiosulphate solution (N2) = ( V I x N 1 ) / V2

= N
Expt. No.: Date:

ESTIMATION OF AMOUNT OF COPPER IN THE GIVEN SOLUTION

BY IODOMETRY

AIM

To estimate the amount of copper present in the given solution by iodometric titration.

PRINCIPLE

Copper ion occurs naturally in drinking water and is a micronutrient required for the
metabolism of living beings. But the presence of copper in water in quantities more than 1.3 mg/L
will cause stomach ache, intestinal distress and digestive problems. High concentration of copper
will also impart a metallic bitter taste to water. Occurrence of copper ions in drinking water may be
due to corrosion in plumbing materials and faulty water treatment processes. Copper is also known
to cause toxicity to aquatic organism.

Titrimetric estimation of copper is done through a redox reaction in which stoichiometric

quantity of iodine is liberated on reaction with potassium iodide. The liberated iodine can be
titrated against standardised sodium thiosulphate.

When KI is added to the copper ion solution, copper ions react with KI and liberates 1 2

2CuSO4 + 4KI 2K2SO4 + 2CuI + I2 

The liberated 12 is titrated against sodium thiosulphate, using starch indicator

2Na2S2O3 + I2 2Nal + Na2S4O6
Sodium tetrathionate

2Cu2+ = I 2 = 2Na2 S2O3

PROCEDURE

Titration I: Standardisation of sodium thiosulphate

20 mL of standard potassium dichromate solution is pipetted out into a clean conical flask.
About 10 mL of dilute H2SO4 and 10 mL of 10% KI are added to it. The liberated iodine is
immediately titrated; against sodium thiosulphate solution taken in the burette. When the solution
turns pale yellow, about 1 mL of freshly prepared starch is added and the titration is continued. The
end point is the disappearance of blue colour. Titration is repeated for concordant values.
 TITRATION 2: ESTIMATION OF COPPER CONTENT IN THE GIVEN SOLUTION

Given copper ion solution Vs sodium thiosulphate solution

S. No. Volume of the Burette reading Volume of Concordant Indicator

given copper ion (mL) sodium thiosulphate value
solution (mL) solution (mL)
Initial Final (mL)

Starch

Volume of sodium thiosulphate solution (V1) = mL

Strength of sodium thiosulphate solution (N1) = N

Volume of the given copper ion solution (V2) = mL

.. Strength of the given copper ion solution (N2) = (V1 x N1) / V2

= N

Calculation of amount of copper present in the given solution “

Amount of copper present in 1000 mL of the solution = Eq. wt. of copper x strength of the given
{ copper ion solution

= 63.5 x N2 (from above)

=
Titration 2: Estimation of copper ion content in the given solution
20 mL of the given copper ion solution is pipetted out into a clean conical flask. One test tube
full of potassium iodide solution is added to this solution and the liberated iodine is titrated against
standardised sodium thiosulphate taken in the burette. When the solution turns pale yellow, about
1 mL of freshly prepared starch indicator is added and the titration is continued. The disappearance
of blue colour is the end point. The titration is repeated for concordant values.

RESULT
1. Strength of the given copper ion solution = N.
2. The amount of copper in the given copper ion solution = g/L.
5
DATA AND CALCULATION

TITRATION 1: STANDARDISATION OF SODIUM THIOSULPHATE

Standard potassium dichromate Vs sodium thiosulphate

S. No. Volume of standard Burette reading Volume of Concordant Indicator

potassium dichromate (mL) sodium value
(mL) Initial Final thiosulphate (mL)
solution
(mL)

Starch

Volume of standard potassium dichromate (V1) = mL

Strength of standard potassium dichromate (N1) = N

Volume of sodium thiosulphate solution (V2) = mL

ç Strength of sodium thiosulphate solution (N2) = (V1 x N1) / V2

= N
Expt. No.: Date:

ESTIMATION OF DISSOLVED OXYGEN IN WATER SAMPLE

BY WINKLER’S METHOD
AIM

To estimate the amount of dissolved oxygen (DO) in water sample by Winkler’s method.

PRINCIPLE
Oxygen can get dissolved in pure water to an extent of 8 nw/L at room temperature. Its
solubility depends on many factors such as temperature, pressure and the chemical and biological
contents of water.
Dissolved oxygen is very essential for the survival of all aquatic lives. A decrease in DO
leads to decay and death of aquatic organisms. The dissolved oxygen (DO) content of a water body
decreases with increase in organic impurity and hence the DO measurements are a good index to
the extent of organic pollution in water.
DO content of water is often responsible for the boiler corrosion. Hence, its determination is
also important in corrosion control.
In the Winkler’s method, dissolved oxygen in the water sample oxidizes Mn2+ ions in
alkaline medium (alkaline KI) to form a brown precipitate of basic manf.4anic oxide (Mn4+).
Mn2+ + 2 OH- Mn(OH)2 
(White)

Mn(OH)2 + ½ O2 MnO.(OH)2 
(DO in water) (Brown)

On acidification, the brown precipitate dissolves and liberates iodine on reaction with F.
MnO.(OH)2 + 4 H+ + 2 I- I +-2 Mn 2+
+3HO 2

The liberated iodine is estimated by titrating with standard sodium thiosulphatc. The amount
of iodine liberated is indirectly equated to the initial DO content of water.

2 S2O32- + I2 S4O6 2- + 2 I-

Sodium thiosulphate is standardised using a standard solution of potassium dichromate. In

acidic medium, dichromate liberates iodine as:

Cr2O72- + 14 H+ + 6 I- 2 Cr3- + 7 H O2 + 3 I 2

The liberated iodine in an amount equivalent to Cr O reacts with sodium thiosulphatc. Starch
2-
2 7
is added as an indicator near the end point. At the end point, the intense blue coloured adsorption
complex of starch with iodine is broken by thiosulphate. The end point is the disappearance of blue
colour, leaving behind a pale green colour due to the presence of Cr3+ ions.
 TITRATION 2: ESTIMATION OF DISSOLVED OXYGEN CONTENT
OF THE GIVEN WATER SAMPLE

Given water sample Vs standardised sodium thiosulphate solution

S. No. Volume of the Burette reading Volume of Concordant Indicator

given water (mL) sodium thiosulphate value (mL)
sample solution (mL)
(mL) Initial Final

Starch

Volume of sodium thiosulpahte solution (VI) = mL

Strength of sodium thiosulpahte solution (N1) = N

Volume of the given water sample (V2) = mL

.•. Strength of the given water sample (N2) = (V1 x N1) / V2

...Amount of dissolved oxygen in the given water sample Strength of the

}( given water sample x 8 x 1000 mg/L

= x 8 x 1000

Amount of dissolved oxygen in the given

water sample } = mg/L
PROCEDURE

Titration 1: Standardisation of sodium thiosulphate

20 mL of standard potassium dichromate solution is pipetted out into a clean

conical flask. About 10 mL of dilute H 2SO4 and 10 mL of 10% KI are added to it.
The liberated iodine is immediately titrated against sodium thiosulphate solution
taken in the burette. When the solution turns pale yellow, about 1 mL of starch is
added and the titration is continued. The end point is the disappearance of blue colour.
Titration is repeated for concordant values.

Titration 2: Estimation of dissolved oxygen in the given water sample

The 250 mL iodine flask is filled with the given water sample up to the
neck. About 2 mL each of alkaline KI and 50% manganous sulphate solutions
are added to the given water sample and the flask is stoppered. Grease is applied
before stoppering and care is taken to see that no air bubble is left inside the flask.
The contents of the flask are mixed by shaking the flask and then allowed to settle
for about 15 minutes. The precipitate is then dissolved by carefully adding 2 mL of
cone. H2SO4 in drops. The contents are mixed well until a clear yellow solution
is obtained. Exactly 100 mL of this clear solution is measured out into a conical flask
using a
measuring jar and titrated against standardised sodium thiosulphate taken in the
burette. When the solution becomes pale yellow, starch is added and the titration is
continued to the end point where the blue colour disappears. A duplicate titration is
carried out.

Equivalent weight of oxygen = 8

RESULT

The amount of dissolved oxygen in the given water sample = mg/L.

4
Expt. No.: Date:

ESTIMATION OF IRON IN THE GIVEN SAMPLE BY

SPECTROPHOTOMETRIC METHOD

AIM

To estimate the amount of iron present in the given sample using spectrophotometer.

PRINCIPLE

Water containing dissolved iron is not suitable for domestic use and industrial applications,
especially in the manufacture of textiles, paper, food additives, drugs and pharmaceuticals. Hence,
the estimation of iron is important in designing the treatment procedure for its removal. Accurate
determination of iron in ppm level cannot be done by titrimetry, conductometry and potentiometry.

Spectrophotometric method of estimation is based on the absorbance of radiation by the

substance in the visible region of electromagnetic spectrum. The intensity of light absorption by a
solution at a specific wavelength is governed by Beer-Lambert's law.

Absorbance A = Log (I0 / I) =  c 1

where, I0 is the intensity of the incident monochromatic light, I is that of the light transmitted
through a solution of thickness 't' and concentration 'c'. '' is a constant characteristic of the
substance and is known as the molar absorption co-efficient. Keeping the path length constant, the
variation in the absorbance with concentration can be measured using a visible spectrophotometer.
The concentration of coloured solutions or solutions which become coloured by the addition of a
reagent can be estimated by this method.

In the estimation of iron, the ferric iron solution of known concentrations is treated with
ammonium thiocyanate reagent to produce a blood red colour due to the formation of an octahedral
ferric thiocyanate complex. The hydrolysis of Fe + solutions is prevented by adding, concentrated
nitric acid.
Fe3+ + 6 NH SCN [Fe(SCN) ]3- + 6 NH +
4 6 4
Blood red
coloured complex

These solutions are subjected to spectrophotometric measurement at a wavelength of 480 nm

and the corresponding absorbance values are noted. A calibration graph is drawn by plotting the
absorbance against the concentration as shown in the fig. (i).
Variation of absorbance of Fe3+ solutions with concentration
measured at 480 nm

Concentration of Fe3+ Absorbance at

solution (ppm) 480 nm
0
2
4
6
8
10
Unknown sample
SPECTROPHOTOMETER

41
PROCEDURE

A stock solution containing 10 ppm of ferric iron is prepared by dissolving accurately 0.0432
g of ferrous ammonium sulphate in 1 mL of concentrated H2SO4 and made up using distilled water
up to 500 mL in a SMF. About 2, 4, 6 and 8 mL of this solution is measured out into 10 mL SMF. To
each of these solutions, 1 mL each of 4N HNO3 and 10% ammonium thiocyanate solution are added
and the contents are made up to 10 mL using de-ionised water. The absorbance of a blank solution
without ferric iron and containing only the reagents is set to be zero in the spectrophotometer. The
absorbance of standard solutions is measured at a wavelength of 480 nm and a working graph is
constructed by plotting the concentrations Vs the corresponding absorbance values.

The given sample of iron solution is mixed with 1 mL each of the reagents and made up to 10
mL. The absorbance of the sample is also measured at the same wavelength and the concentration
of Fe3+ is determined from the graph.

RESULT

The amount of iron present in the given sample = ppm.

42