Chemistry Lab-I: Atomic Structure, Bonding, General Organic Chemistry and Aliphatic Hydrocarbons

BCHCL-132
CHEMISTRY LAB-I:
Indira Gandhi National
Open University ATOMIC STRUCTURE, BONDING,
School of Sciences
GENERAL ORGANIC CHEMISTRY
AND ALIPHATIC HYDROCARBONS
BCHCL-132
Indira Gandhi National
CHEMISTRY LAB I:
Open University ATOMIC STRUCTURE, BONDING,
School of Sciences
GENERAL ORGANIC CHEMISTRY
AND ALIPHATIC HYDROCARBONS
UNIT 1
Titrimetry: An Introduction 7
EXPERIMENT 1
Determination of Sodium Carbonate and Sodium
Hydrogen Carbonate Present in a Mixture 30
EXPERIMENT 2
Estimation of Oxalic Acid by Redox Titration 43
EXPERIMENT 3
Estimation of Water of Crystallisation in Mohr’s Salt 56
EXPERIMENT 4
Estimation of Iron(II) Ions by Chromatometry using
Internal Indicator 63
EXPERIMENT 5
Estimation of Copper(II) Ions Iodometrically 70
EXPERIMENT 6
Detection of Extra Elements (N, S, X) in the Organic
Compounds 80
EXPERIMENT 7
Separation and Identification of the Components of a
given Mixture of Amino Acids by Paper Chromatography 92
EXPERIMENT 8
Separation and Identification of the Sugars Present in
the given Mixture by Paper Chromatography 106
Course Design Committee
Prof. H. B. Singh (Retd.) School of Sciences,

Dept. of Chemistry, IGNOU
University of Delhi,
Delhi Prof. Vijayshri
Prof. A.K. Bakhshi
Prof. Sunita Malhotra
Dept. of Chemistry,
University of Delhi, Prof. Javed A. Farooqi
Delhi
Prof. Amir Azam Dr. Lalita S. Kumar
Dept. of Chemistry,
Jamia Millia Islamia,
New Delhi
Prof. Tabrez Alam
Dept. of Chemistry,
Jamia Millia Islamia,
New Delhi
Prof. J.M. Khurana
Dept. of Chemistry,
University of Delhi,
Delhi
Block Preparation
Prof. Bharat Inder Fozdar Prof. B.S. Saraswat (Retd.) (Editor)
School of Sciences, IGNOU School of Sciences, IGNOU
Prof. Lalita S Kumar
School of Sciences, IGNOU
Course Coordinator Prof. Bharat Inder Fozdar and

Prof. Lalita S Kumar
Production
Mr. Sunil Kumar
Assistant Registrar (Pub.)
Acknowledgements: Sh. Sarabjeet Singh and Sh. Deepak Kumar for word processing and
Sh. Sarabjeet Singh for CRC preparation.
The utilisation of some content of Chemistry Lab I (CHE 03L) & III (CHE 08L) courses is gratefully
acknowledged.
July, 2019
 Indira Gandhi National Open University, 2019
ISBN: 978-93-88980-98-2
All rights reserved. No part of this work may be reproduced in any form, by mimeograph or any other
means, without permission in writing from Indira Gandhi National Open University.
Further information on Indira Gandhi National Open University courses may be obtained from the
University’s office at Maidan Garhi, New Delhi-110 068 or IGNOU website www.ignou.ac.in.
Printed and published on behalf of Indira Gandhi National Open University, New Delhi by
Prof. M.S. Nathawat, Director, School of Sciences.
Printed at: Chandra Prabhu Offset Printing Works Pvt. Ltd. C-40, Sector 8, Noida-201 301, U.P.
CHEMISTRY LAB I: ATOMIC STRUCTURE,
BONDING, GENERAL ORGANIC CHEMISTRY AND
ALIPHATIC HYDROCARBONS
Welcome to the first laboratory course in chemistry. This manual gives the procedural details
of various experiments you are required to perform in the course of this lab work. Appropriate
conceptual basis has also been laid for each experiment to make this manual complete in
itself.
Two common tools, a scientist uses, are observation and reasoning. An experiment is a
controlled observation. We perform an experiment having some aim in our mind. We make
observations and draw conclusions by logically analysing these observations. Chemistry has
been defined as ‘the integrated study of the preparation, properties, structure and reactions
of the chemical elements and their compounds and of the systems which they form’. As such
experimental work forms an essential part of chemistry which has, indeed, been defined as
an experimental science. One type of experiments in chemistry is known as analytical
experiments. These deal with analysing various materials to find the nature and the amount
of components constituting them. The other type of experiments deals with the synthesis of
substances.
Broadly, analytical experiments are further divided into two types, viz., qualitative,
concerned with the identification and separation of chemical substances, and quantitative,
concerned with the determination of the amount of a chemical substance present, either
alone or in a simple or complex mixture with other substances. Quantitative analysis is
further classified into various types, e.g., gravimetric analysis, volumetric or titrimetric
analysis, etc.
In this course we begin by explaining the basic concepts involved in titrimetric analysis under
the quantitative type. In Unit 1 you would be introduced first to the basic skills, such as, how
to weigh a sample, measure volumes and prepare a solution for titrimetric analysis. Perhaps
you might have learnt these skills in your previous classes. This unit is an important review
and extension. You will also learn the handling of lab reagents and safety measures to be
observed while working in the laboratory. After the unit there are a number of quantitative
experiments given with procedural details. The 1st experiment is based on a neutralisation
type of titration. Experiment 2, 3 and 4 are based on redox titrations and the 5th experiment is
based on iodometry.
Under the qualitative type of analysis you will detect extra elements in organic compounds in
Experiment 6. Experiment 7 and 8 deal with a separation technique called paper
chromatography where you will separate and identify components in a given mixture of
amino acids and a mixture of sugars.
Expected Learning Outcomes
After studying this course and performing the experiments set in it you will be able to:
• explain the basic concepts involved in titrimetric analysis,
• explain the principles of acid-base, redox and iodometric titrations,
• work out the stoichiometric relations based on the reactions involved in the above,
• take observations and calculate results after performing the experiments,

• carry out the physical examination of organic compounds,
• determine the elements (N, S, X) present in the given organic compound, and
• separate and identify the mixture of compounds by paper chromatography.
Study Guide
In this lab course, you would be doing eight experiments that involve the qualitative as well
as the quantitative analytical techniques.
Write up on each experiment starts with a discussion of the theoretical principles on which
the experiment is based. We would like you to go through this carefully before starting the
experiment. We have introduced Self Assessment Questions (SAQs) at appropriate
places, which will enable you to see whether you have understood this part.
The section/sub-section, ‘Requirements’ gives you an idea of the apparatus and chemicals
you would need for the experiment. You may have to prepare some solutions yourself, for
which detailed procedures have been given. In addition, some solutions may be provided,
which you may collect from the laboratory staff.
The next three section/sub-sections, i.e., ‘Procedure, Observations and Calculations’ tell
you how to carry out the experiment, take observations, tabulate them and calculate the
result. In the last section/sub-section, ‘Result’, you are required to discuss your result and
compare it with the known value, your counsellor will give you.
With the result for each experiment you have to mention % error which you can calculate
using the following formula,
Experimental Value − Correct Value

% Error = × 100
Correct Value
We want you to see where the source(s) of error are, so that you can improve your
performance. For example in a titration experiment, possible explanation for the experimental
low values for the molarity include (a) error in weighing of primary standard, (b) in the
preparation of standard solution, (c) in taking the burette readings, etc.
Last but not the least, check the apparatus as it is given to you, especially the burette, it
should not leak. Clean the glass apparatus thoroughly as indicated. After you finish the
experiment, clean the apparatus again. You should leave your bench as clean as you got it.
We have given elementary precautions that need to be observed in the laboratory. Read
them carefully and observe them. Mind you, chemicals are safe if handled properly.
Laboratory Notebook
An important part of scientific training is the maintenance of a complete and up to date record
of your laboratory work. For recording experimental data, laboratory notebooks are available
in the market. Purchase a 30-40 page chemistry notebook for this laboratory course.
You should prepare the pages for recording data before you come to the lab. For each
experiment, you should write down the title of the experiment, important chemical reactions
involved and observations. The observation tables, as given with the experiment in your
manual, should be given on the left-hand page. Calculations and results are reported on the
right-hand page. You may do the calculations after the lab and record the result in your note
book.
The laboratory notebook must be submitted to the counsellor for corrections and grading.
Marks have been allocated for doing the experiment and for recording it properly.
Your counsellor may also conduct a viva-voce to judge how much you have learnt. There are
marks for this too.
We want you to share the thrill of learning by doing. There is no better way to learn.
So, Best of Luck.

IMPORTANT
• Attendance is compulsory in the Laboratory Course work held generally
at the Study Centre.
• The Laboratory Course is worth 2 credits to be completed over 7 days
duration:
6 days of Guided Laboratory work
1 day for the Unguided Laboratory work
• To successfully complete the laboratory course you will have to pass (at
least 35% marks) in the Guided and Unguided components separately.
Unit 1 Titrimetry: An Introduction
UNIT 1
TITRIMETRY:
AN INTRODUCTION
Structure
1.1 Introduction 1.4 Standard Solution
Expected Learning Outcomes 1.5 Titration
1.2 Apparatus Commonly Used Types of Indicators
How to Use a Pipette Types of Titrations
How to Use a Burette 1.6 Instrumental Determination of
How to Use a Volumetric Flask Equivalence Point
How to Use an Analytical Balance 1.7 Safety Measures in the
Single-Pan Electronic Analytical Laboratory
Balance 1.8 Answers
1.3 Expression of Concentration
1.1 INTRODUCTION
In the very first laboratory course of the first semester, there are five
experiments based on the titrimetric analysis besides the elemental analysis of
organic compounds and experiments based on chromatography. The
introductory unit of the laboratory course has been designed to familiarise you
with the apparatus which will be used in performing the titrimetric experiments
and some of the basic concepts on which the experiments are based.
You might have studied in your previous classes that in titrimetry we estimate a
substance in solution by titrating it against the standard solution of an
appropriate substance. The whole process involves the use of specific
apparatus that you need to be thoroughly familiar with. Therefore, first of all we
introduce you to the apparatus commonly used in titrimetric analysis, and
explain its correct use. We also tell you how to make a standard solution and
express its concentration. The weighing of chemicals being a part and parcel of
these experiments, the correct use of analytical balance has been dealt in
detail. Finally, we introduce you to the common safety measures one should
observe in a Chemistry laboratory.
7
BCHCL-132 Chemistry Lab I

After studying this unit, you should be able to:
measure and deliver sample volumes by selecting and using appropriate

apparatus for titrimetric measurement;
determine the mass of a sample by correctly using analytical balance;
perform basic laboratory skills, including pouring reagents and transferring

solids, preparing solutions of known concentrations;
list and explain different types of titrations; and
explain the safety measures as applied to a chemistry laboratory.
1.2 APPARATUS COMMONLY USED

Titrimetric analysis involves reliable and accurate measurement of volumes of
solutions. Three pieces of apparatus, namely, a pipette, a burette and a
volumetric flask are indispensable for this purpose. Their use is described
here. Before doing the experiment you should go through the instructions
given below carefully and work accordingly.
Pipettes which can 1.2.1 How to Use a Pipette

measure volumes of less
3
than 1 cm are also Pipette is used to measure and transfer known volume of a liquid from one
available with special container to the other.
accessories.
A pipette is shown in Fig. 1.1 (a). As you can see, it is a long tube with a bulb
in the middle. On the narrow upper part of the pipette a horizontal line is
marked. This line indicates the level to which the pipette has to be filled to
deliver the liquid equal to the volume indicated on the bulb when used in the
way described below. Pipettes can be of different capacities like 1, 2, 5, 10,
20, 25, 50 cm3, etc. You will use pipettes mostly of 10 and 20 cm3 for your
experiments.
CAUTION!
Do not suck corrosive
liquids like strong acids
and alkalies by mouth.
You can use a rubber
teat for this purpose.
Fig. 1.1: (a) Pipette. (b) Handling of a pipette. (c) Correct way to drain
8 out the solution.
Before using a pipette, it has to be thoroughly washed with a good quality The curved surface of a
detergent followed by plenty of water and finally with distilled water. This liquid in a container is
removes all the grease. It is then rinsed with the solution which has to be known as the meniscus.
measured. For rinsing, the solution is taken in a clean and dry beaker. The The meniscus in case of
pipette is dipped deep into the solution and the solution is sucked into the liquids which stick to the
pipette to fill it up to about half its volume. It is then taken out and the solution container, is concave,
e.g. for water and
is made to wet it completely from inside by moving the solution up and down
aqueous solutions, while
and also around its axis. The solution is drained out and the whole process is it is convex in case of
repeated. The pipette is then filled with the solution until the level is about liquids which do not stick
2 cm above the mark. The top of the pipette is then quickly closed by slightly to the container, e.g. for
moist (not wet) index finger; see Fig. 1.1 (b). The pressure of the finger is mercury.
slowly released so as to allow the solution to run out until the lower meniscus
just touches the mark. The solution from the pipette is transferred into the
container in which titration has to be done. The solution is allowed to run out
on its own. The last drop of the solution which does not seem to drain out by
itself is taken out gently by touching the tip of the pipette with the walls of the
container for about 3-4 seconds; see Fig. 1.1 (c). Do not blow out the last
drop. The pipette is calibrated to include the liquid column trapped at the tip.
Further, blowing makes it dirty and CO2 in the breath may react with the
solution being pipetted. The volume of the liquid thus transferred through the
pipette is equal to the volume marked on the pipette.
Another type of pipette is designed to deliver definite but different volumes of a

liquid. It is called a graduated pipette, Fig. 1.2. It has got marking
corresponding to different volumes. It is also used in a similar fashion, with the
only difference that the liquid is not completely drained out; instead the volume
required is transferred.
SAQ 1
Why should you not blow the last drop out of the pipette? Fig. 1.2: Graduated
Pipette.
1.2.2 How to Use a Burette

A burette is designed to transfer definite but variable volumes of a liquid into
another container.
A burette is a long glass tube, commonly of 50.0 cm3 capacity in 0.1 cm3 unit
graduation marks, Fig. 1.3. It has a stop cock at the lower end to control the
amount of solution drained. The burette also has to be washed, first with a
detergent followed by plenty of water and finally with distilled water. It is then
rinsed with the solution to be measured. For rinsing it is filled a little less than
half with the solution and by repeatedly rotating and tilting the burette, the
solution is made to wet it completely from inside. This solution is discarded.
The burette is then mounted on the stand in an upright position and is filled
carefully with the help of a funnel. After taking out the funnel, the meniscus is
adjusted to a definite graduation mark by drawing out some solution through
the stop cock. The bottom of the meniscus should just touch the graduation
mark. While reading the solution level in the burette, your eyes should be on
level with the graduation mark, otherwise there would be error due to parallax,
Fig. 1.4. It is not necessary to adjust the meniscus at the zero mark level, if it 9
is too high for the level of your eyes. You can adjust it at, say 10.0 cm3 or any
other convenient level.
Error in burette reading is among the most common sources of error in

titrimetric analysis. To make the meniscus more distinct and to ensure that it
looks the same always, it is convenient to place a screen behind the burette as
shown in Fig. 1.5. This can be made from a small piece of cardboard covered
with white paper with the lower half blackened with ink. The black part is to be
held downward. This is called a parallax card. You can ask your counsellor to
show you how to make a parallax card.
Fig. 1.3: Burette. Fig. 1.4: Eye level for Fig. 1.5: Reading the burette with
burette reading. the use of the parallax
card.
After adjusting the meniscus, the level of the solution in the burette is
recorded. This is called the initial reading or initial volume. Then the titration
PRECAUTION! is performed and at the end of the titration, the level of the solution is
recorded. It is called the final reading or final volume. The difference of the
No standard apparatus
two readings, (final reading – initial reading), gives the volume of the solution
is to be heated above
298 K. transferred to the titration flask. The correct way of delivering a liquid from
burette is shown in Fig. 1.6.
10 Fig. 1.6: Delivery of liquid from a burette.

1.2.3 How to Use a Volumetric Flask
A volumetric flask is used to prepare a definite volume of a solution of
precisely known concentration.
Volumetric or measuring flask has a flat bottom with a long, narrow neck,
Fig. 1.7. It has a calibration mark on its neck which indicates the level up to
which the flask is to be filled to get a volume equal to the one indicated on the
flask.
Fig. 1.7: Volumetric Flasks.
You will be using volumetric flasks of 100 cm3 and 250 cm3 capacity. The
flask, before use, is cleaned thoroughly, washed with a detergent and plenty of
distilled water and allowed to drain. The weighed compound is transferred into
the flask with the help of a funnel. It is first dissolved in just enough water; the
solution is then made up to the mark by carefully adding more distilled water.
This can be done with a wash bottle or better with a pipette. The flask has to
be stoppered tightly and shaken well before use to get a homogeneous
solution.
1.2.4 How to Use an Analytical Balance

In titrimetric analysis, you will invariably have to prepare a standard solution.
You would be required, for this purpose to weigh a solid accurately by using
an analytical balance. It is very important to learn the use of an analytical
balance because accurate weighing is important for the accuracy of any
titrimetric experiment.
Generally two types of analytical balances are used in chemical laboratories.

These are:
(i) Double pan analytical balance
(ii) Single pan electronic analytical balance
Double pan analytical balance is now outdated but still some of our Zero point is the point
laboratories do not have electronic analytical balance and may have only on the scale at which the
double pan balance. A commonly used double pan analytical balance is pointer of the unloaded
shown in Fig. 1.8. The various parts of the balance are labeled in the figure. balance comes to rest.
Before using the balance, you have to first determine the zero point of the
balance. For this purpose, the side doors of the balance are closed and the
arrest knob (1) is slowly and carefully turned counter-clockwise. Avoid jerks
as they may disturb the setting of the balance. 11
When the arrest knob is turned fully to the left, the pointer (2) starts swinging
around the centre of the scale (3). The first two swings are ignored and
starting with the third swing, the extreme positions of the swing are noted. The
swings to the right are positive and those to the left are negative.
Fig 1.8: Analytical balance.
The readings to the left and right are averaged separately and the mean of
these averages is found, which is the zero point. The following example will
make it clear.
Ideally the zero point and
the mid or zero of the
scale should be the Reading on the left Reading on the Right
same.
1. – 5.0 + 5.0
2. – 4.0 + 4.0
3. – 3.0 + 3.0
4. – 2.0
– 14.0 + 12
− 14.0 12.0
Average = = −3.5 = 4.0
4 3
− 3.5 + 4
Mean Value = = 0.25
2
The zero point is + 0.25, i.e. 0.25 units to the right.
Such small discrepancies between the zero point and the middle of the scale
12 may be ignored as they are insignificant. However, if the deviation is large,
e.g., greater than 1.5 units, the balance must be adjusted by means of the
screws (4), for which you may request your counsellor.
After adjusting the zero point of the balance (if necessary), we come to actual
weighing. For this purpose, we use a glass or a plastic weighing bottle,
Fig. 1.9. First of all, the weighing bottle is weighed on a rough balance to find
its approximate mass to the nearest gram. Then, the left side door of the
analytical balance is opened and the weighing bottle is kept on the left side
pan (5) and the door is closed. Similarly, through right side door, weights equal
to the approximate mass of the weighing bottle are transferred to the right side
pan from a weight box; Fig. 1.10.
Fig. 1.10: Weight box and weights.
You must close both the doors of the balance before raising the pans with the arrest knob.
The arrest knob is once again turned to the left and the movement of the
Always use forceps to
pointer is seen. If it moves more to the left, then the weights transferred are in transfer the weights.
excess of the mass of the bottle. In that case some weights have to be Refrain from using
removed. On the other hand, if the pointer moves to the right, then the added your hands.
weights are not sufficient and we need to add more weights. Arrest the
movement of the beam by turning the arrest knob fully towards the right and
open the right side door to add or remove some weight(s), as the case may
be. Recheck the movement of the pointer by turning the arrest knob. Continue
this process till the addition of 1 gram weight makes the right hand pan heavier
while its removal makes it lighter, e.g., if the weight is say 15.5 g, then 15 g
weight would be lighter and 16 g weight would be heavier. After this, the
fractional weights marked in mg, have to be added in the order of decreasing
weight till the two sides are balanced. Do not use fractional weights of less
than 10 mg, you should use a rider in such cases. A rider, Fig. 1.10, is a thin
metallic wire suitably bent to be seated on the beam of the balance. It is
.
normally put on the right hand side of the beam (6) with the help of the rider
carrier (7). By varying the position of the rider on the beam (8), the rest point is
found, i.e., the two pans are balanced.
The beam scale has got markings from 0-10 on either side. It is calibrated in
such a way that each main division is numerically equal to mass in milligram,
when the rider is put on it. Each main division is further divided into 5
subdivisions and each subdivision is equivalent to 0.2 mg. thus the accuracy
of such an analytical balance can be only up to 0.2 mg. The mass of an object
can be calculated using the following formula:
13
Mass of the object = (Weights added in grams) + (Fractional weights added ×
0.001) g
+ (Main division of the rider position × 0.001) g
+ (Subdivision of the rider position × 0.0002) g
Let us illustrate the use of this formula. Suppose that while weighing an object,
the weights added to the right side pan are 15 g, 200 mg and 2 × 20 mg. Let
the rider position be 3 on the subdivisions after 2 main divisions.
Then the mass of the object
Mass of the substance = 15.00 g + (240 × 0.001) g + (2 × 0.001) g + (3 × 0.0002) g = 15.2426 g

(m g) = Mass of the
bottle with substance You have, so far, seen how to weigh an object accurately. If we want to weigh
(m1 g) − Mass of the substance in the weighing bottle, we make use of the method of weighing by
bottle after transferring
difference. For this, the weighing bottle is first approximately weighed. The
the substance (m2 g)
m = m1 – m2 g substance to be weighed is put into the bottle (a little more than required) and
weighed accurately (m1 g). The substance is transferred into a volumetric flask
and the bottle is again weighed accurately (m2 g). The difference of the two
masses, i.e., (m1 − m2) gives the exact amount of the compound transferred
(m g).
1.2.5 Single-Pan Electronic Analytical Balance

In case of electronic balance also known as digital balance, the mass of the
object being weighed will be digitally displayed. In electronic balances
(Fig.1.11), substance can be added to or removed from the balance without
any problem.
Handling of Electronic Balance
Balances of all types should never be subjected to harsh treatment of any

description. Always make adjustments smoothly and carefully. You will need to
ensure that the balance reads zero before you start to weigh. However, if you
are using a balance with a tare facility, it is not necessary to zero the balance.
14 Fig.1.11: Single-pan electronic analytical balance.

The tare facility allows you to cancel out the mass of the container so that you
can weigh the required mass of material without having to take into account
the mass of the container. Even if you cannot tare out the complete mass of
the container, the tare facility can still be used to adjust the mass indicated by
the scale to a convenient whole number.
For example, suppose you are using a balance that can only tare out 10 g, but
your container weighs 22.45 g and you require to weigh 8.70 g of material. In
such a case instead of
(1) weighing the container,
(2) adding its mass to the required mass of material, and
(3) adding material to the container until the balance reads the calculated
mass,
you can use the tare facility to adjust the displayed mass of the container to
read 20.00 g and then add material to the container until the balance reads
28.70 g. This avoids errors in weighing that arise due to miscalculations! Of
course, when you have finished weighing, you should remove any tare and
adjust the balance to read zero.
An important point to note here is that you should always use a container for
weighing any material. Even a piece of filter paper is not very satisfactory as Never weigh anything
solid chemicals can easily spill onto the balance pan. As already mentioned (apart from the
small glass or plastic weighing bottles are available and these are most useful. container) directly on the
balance pan.
For very accurate weighing, e.g. in the preparation of a standard solution it
may be necessary to use a weighing bottle.
Having learnt about the general apparatus to be used in the experiments for
the first laboratory course, let us now understand the various terms and
concepts used in these experiments. Before this, try to answer the following
SAQ.
SAQ 2
What is the mass of a substance if the following weights are needed to weigh
it?
g mg position of rider
5 200 8.2
2 100
1 50 Solute is the dissolved
substance in a solution.
Solvent is the liquid in
1.3 EXPRESSION OF CONCENTRATION which the solute is
dissolved. Solution is
In a qualitative sense, the term concentration deals with the “crowdedness” of the homogeneous
mixture of a solute and a
the particles of solute in a solution. A solution having more number of solute
solvent.
particles per unit volume is said to be more concentrated. In quantitative
analysis, one very often comes across this term. Before we give an expression
for this, it would be worthwhile to recapitulate a few relevant fundamental
concepts here. 15
Mole, denoted as mol, is the amount of a substance that contains as many
elementary entities as are there in 0.012 kg of C12 isotope of carbon. The mole
may be of atoms, ions, molecules, electrons or any other entity. The number of
12
The number of C atoms elementary entities in a mole of any substance is fixed and is given by a
12
in 0.012 kg of C is constant called the Avogadro’s number, NA which equals 6.022 × 1023.
23
equal to 6.022 × 10 .
Relative Molecular Mass (Molecular Weight) denoted as Mr, is the mass of
one molecule in atomic mass unit (a.m.u.) relative to 1/12th of the mass of the
pure C12 isotope (12.000 a.m.u.). For most titrimetric analyses, purpose of this
Relative molecular mass is the same as the old atomic mass and molecular mass. We find it by
being relative is unitless. multiplying the atomic mass of each element in the molecule by its subscript in
the formula and then adding the total for each element to get the grand total,
e.g., one molecule of CO2 has relative molecular mass of 44, which is
calculated as:
Although SI unit of molar [12 + (16 × 2)] = (12 + 32) = 44
-1
mass is kg mol , it is
more convenient to use g Molar Mass, denoted by symbol Mm, is the mass of one mole of a given
-1
mol for titrimetric substance. It is numerically equal to the relative molecular mass but is
calculations.
expressed in g mol-1 units. The following illustration explains this point.
The relative molecular mass of oxalic acid dihydrate [(COOH)2.2H2O] crystals

= 126. The molar mass of oxalic acid dehydrate crystals = 126 g mol-1.
The amount of a substance having mass equal to molar mass is called a

mole. Thus we see that mole, molar mass and Avogadro’s number are
interrelated. A schematic representation of the relationship among these is
shown below:
1 Mole 1 Molar mass
23
6.022 × 10 entities
For titrimetric purposes we express concentration in terms of molarity

denoted by symbol M which is defined as the number of moles present in
one dm3 of the solution. It can be expressed as:
Number of moles of solute

Molarity (M ) =
Volume of solution (in dm 3 )
Thus, if you dissolve 126 g of oxalic acid dihydrate (molar mass =126 g mol−1)
in water and make the volume up to 1 dm3, then the solution would be 1M.
The molarity, M, of a solution containing m g of the solute in V cm3 of a

solution can be calculated as follows:
Let the molar mass of the solute be Mm g mol−1
Its mass
Number of moles of a solute =
Molar mass
m
g m
Mm
= = mol
−1 Mm
16 g mol
3
Volume of the solution = V cm
Since 1000 cm 3 = 1 dm 3 ,
V cm 3 V
Volume of the solution = = dm 3
3 −3 1000
1000 cm dm
Number of moles of the solute

Hence by definition , Molarity of the solution (M ) =
Volume of the solution in dm 3
m Mm
1
V
= mol ×
dm 3
1000
mV
M
1000
M
i.e., = mol dm−3 (1.1)

.
The pipettes and burettes are calibrated in cm3 units (actually in ml which is 3
1 ml = 1.000028 cm
almost equal to cm3). Hence, by substituting the volumes of the solutions
in cm3 units in the above expression, molarity of a solution can be
calculated.
Though molarity is being accepted more and more as the way of expressing
concentrations, another related term, viz., normality is still in use. Here
equivalent mass is used in place of molecular mass. Normality is defined as
the number of gram equivalents of the solute per dm3 of the solution. In other
words,
Number of equivalens of solute
Normality (N) =
Volume of solution (in dm3 )
The molar mass of a substance is an inherent property. It is independent of

the nature of the chemical reaction it may be undergoing. Hence, a given
solution containing a known amount of the solute will have the same molarity
under all conditions. Normality, on the other hand, can change as the gram
equivalent of a substance depends on the chemical reaction involved in the
titration. For example, KMnO4 can have a gram equivalent of 158.04, 52.68 or
31.6 depending on the reaction conditions. In the light of the above, it is
advisable to use molarity rather than normality. We would be using molarity
throughout our experiments. However, percentage, formality, molality, mole
fraction and ppm are some other ways of expressing concentration and are
briefly explained here.
Percentage: The percentage of a solute in a given solution can be expressed

in three different ways depending upon the nature of the solute and the
solvent. Let us illustrate by taking some examples.
(a) If we take 10 g of, say, NaCl and dissolve it in water to make a volume of
100 cm3, then we get a 10% mass by volume, i.e. 10% m / V solution of Mass per cent can also
NaCl in water. be called parts per
hundred (pph).
(b) If instead of preparing 100 cm3 of solution, we add enough water to
prepare 100 g of solution, then we get 10% mass by mass, i.e. 10% m / m
solution of NaCl in water. 17
(c) In cases where the solute is also a liquid, it is possible to represent

concentration as volume by volume. For example, if we mix 10 cm3 of
methanol (solute) with H2O (solvent) to prepare 100 cm3 of the solution,
then we get 10% volume by volume, i.e. 10% V / V solution of methanol in
water.
Mathematically percentage is given as:
Amount of solute
Percentage = ×100
Amount of solution
The units would depend on the units of the amount of solute and solvent.
Formality: In certain ionic compounds, e.g., NaCl, which are completely

dissociated in solution, it is less accurate to talk of one molecule or of
molecular mass. In such cases, a different term, viz., formality is considered.
Formality is defined as the number of gram formula masses dissolved per dm3
of the solution. Here, it is, therefore, more appropriate to talk of formality than
of normality or molarity.
Molality: The molality of a solution is the number of moles of the solute per
kilogram of the solvent contained in a solution. It is given by the following
expression:
m1 ×1000
Molality = m × M
2 m
m1 = mass of the solute
m2 = mass of solvent
Mm = Molar mass of the solute
The molality scale is useful for experiments, in which physical measurements,

e.g., freezing point, boiling point, vapour pressure, etc., are made over a wide
range of temperatures.
Mole fraction: The mole fraction (x) of any component in a solution is defined
as the number of moles (n) of that component divided by the total number of
moles of all the components in the solution. The sum of mole fractions of all
the components of a solution is unity. For example, for a two component
solution, the mole fraction can be given as follows:
N
x
n
N
1
(solvent) =
+
n
x
n
N
2
(solute) =
+
nn
NN
x
x
+
1
+ 2
= =1
+
Here n is the number of moles of the solute and N is that of the solvent. Mole
fraction scale is mostly used in theoretical work.
18
Parts per million (ppm): This unit is particularly useful for expressing very
1 mg mass of a solute
small concentrations. We find this unit by using: 3
dissolved in 1 dm is one
mass of solute ppm.
ppm = ×1,000,000 -3
mass of solute + mass of solvent ppm = mg dm
-3
= µ g cm
The masses of solute and solvent should be expressed in the kg unit. The
-3 -3
concentration of air and water pollutants are often given in parts per million. = 10 g dm
Various ways of expressing concentrations are given here just to make you
aware of these. Though in modern texts, by and large, the concept of molarity
is being used, you would come across other expressions also.
SAQ 3
What is the molarity of sodium hydroxide solution made by dissolving 4.000 g
of solute in a volumetric flask and adding water to the calibrated volume of
500 cm3? (Mm of NaOH = 40 g mol-1).
SAQ 4
How many grams of AgNO3 will have to be weighed to make 1 dm3 solution of
0.1 mol dm-3 molarity? (Mm for AgNO3 = 169.87 g mol-1).
1.4 STANDARD SOLUTION

The concentration terms being clear to you, you must know about a standard
solution which is a significant part of all the titrimetric experiments.
A standard solution is defined as the one whose concentration (strength) is

known accurately, i.e., we know exactly how much of the solute is dissolved in
a known volume of the solution. A standard solution may be prepared by
dissolving an accurately weighed, pure stable solid (solute) in an appropriate
solvent. Preparation of a standard solution is generally the first step in any
quantitative experiment, so it is important to know how to prepare a standard
solution.
Primary and Secondary Standards
In titrimetry, certain chemicals are used frequently in defined concentrations as

reference solutions. Such substances are classified as primary standards or
secondary standards. A primary standard is a compound of sufficient purity
from which a standard solution can be prepared by weighing a quantity of it
directly, followed by dilution to give a definite volume of the solution. The Hygroscopic
following specifications have to be satisfied for a substance to qualify as a substances are those
primary standard: which have a tendency
to absorb moisture.
1. It must be easily available and easy to preserve.
2. It should not be hygroscopic nor should it be otherwise affected by air.
3. It should be readily soluble in the given solvent. 19

4. The reaction with a standard solution should be stoichiometric.
5. The titration error should be negligible.
Few available primary standards for acid-base, redox and complexometric

titrations are given in Table 1.1.
Table 1.1: List of primary standards used in acid-base, redox and

complexometric titration
Name of the primary Formula Molecular Type of

standard mass titration
Potassium hydrogen (KHP) C8H5O4K 204.23 Acid-base

phthalate
Anhydrous sodium Na2CO3 106 Acid-base

carbonate
Potassium dichromate K2Cr2O7 294.19 Redox
Arsenic (III) oxide As2O3 197.85 Redox
Potassium iodate KIO3 214.00 Redox
Sodium oxalate Na2C2O4 134.00 Redox
Sodium Salt of EDTA C10H16N2O8 372.3 Complexometric
Solutions prepared from the primary standards are called primary standard
solutions.
Substances which do not satisfy all the above conditions are known as
secondary standards. In such cases a direct preparation of a standard solution
is not possible. Examples are alkali hydroxides and various inorganic acids.
These substances cannot be obtained in pure form.
Therefore, concentration of these can be determined by titrating them against

primary standard solutions. This process is called standardisation and the
solution so standardised is called a secondary standard solution.
Preparation of a Standard Solution
To prepare a standard solution of volume, V cm3, of known molarity,

M mol dm-3, the mass of the solute required, m g, of molar mass Mm, can be
calculated by rearranging Eq. 1.1 as follows:
M . Mm . V
Mass of the solute (m) = g (1.2)
1000
The solute is then weighed on an analytical balance as explained before

(Sec 1.2.4), transferred into a standard flask and dissolved first in a small
20
quantity of the solvent, the solution is then made up to the mark and shaken
thoroughly to get a homogeneous solution.
In preparing a standard solution whose concentration is, say, around 0.1 M,

the amount of the substance weighed need not be exactly equal to that
corresponding to 0.1 M. It can be slightly less or more, but the weighing must
be accurate. From the weight of the solute actually taken, molarity of the
solution can be calculated using Eq. 1.1.
SAQ 5
According to the criteria given in the text, on what criteria do (a) sodium
hydroxide and (b) benzoic acid fail as primary standards?
1.5 TITRATION
In titrimetric analysis, one determines the volume of a standard solution which Aliquot is the volume of
is required to react quantitatively with a known volume of the other solution, the solution delivered by
the concentration of which is to be determined. For this purpose, an aliquot of the pipette in a titration. If
3
the solution to be estimated is pipetted out and is transferred to a conical flask. you use a 20 cm pipette
The standard solution is added dropwise from a burette to the solution in the every time during a
conical flask. The conical flask is continuously shaken to enable the two titration, the aliquot
3
contains 20 cm of the
solutions to mix thoroughly. Standard solution is added till the two solutions
solution.
react quantitatively. This process is called titration. The solution in the conical
flask is called the titrand and the one in the burette is called the titrant. The
total volume of titrant used in the reaction is called the titre. The steps in a
titrimetric estimation are depicted in Fig. 1.12.
We have stated above that in a titration, the titrant is added till it reacts
quantitatively with the titrand. Such a stage, at which the quantities of titrant
and titrand are in their stoichiometric proportions (in terms of equivalents or
moles), is called the equivalence point. A question arises now, as to how do
we know that the equivalence point has been reached? At what stage shall we End point is the point
usually indicated by a
stop adding the solution from the burette? Essentially we need some
change of colour of an
substance which can indicate this stage by a change in a physical property like
indicator. At the end
colour. A substance which is used to indicate the equivalence point of a point particular reaction
titration through a colour change is called an indicator. Equivalence point so is completed.
obtained is called end point. It is not necessary that the end point is Equivalence point is the
coincident with the equivalence point, because of the delay in getting the point at which the
indicator to show the change, and other factors. Ideally end point and number of equivalents of
equivalence point should be as close as possible. The indicator, to be used in reactants are equal to
each other.
a given titration, would depend on the nature of the chemical reaction involved
between the two reacting solutions. The basic requirement for an indicator is
that it should have distinctly different colours before and after the end point
because we need to know the end point visually. If no visible indicator is
available, the detection of equivalence point can often be achieved by
following the course of the titration by measuring the potential difference
between an indicator electrode and a reference electrode or the change in the
conductivity of the solution.
21
Fig. 1.12: Steps in a titrimetric estimation.
1.5.1 Types of Indicators

The indicators can be of three types depending upon their usage:
i) Internal indicators: These have to be added into the reaction solution.

Examples are: phenolphthalein, methyl orange, diphenylamine, etc.
ii) External indicators: These are not added into the solution. The
indicator is kept out on a plate. A drop of the solution being titrated is
taken out with the help of a rod and put on the indicator. A change in
colour indicates the end point. Potassium ferricyanide is one such
example.
iii) Self-indicators: Sometimes either the titrand or the titrant changes its
colour at the end point and acts as a self-indicator. The example is
potassium permanganate used in permanganatometry.
1.5.2 Types of Titrations

Depending upon the nature of the chemical reaction involved in a titration, the
latter can be classified into the following types:
i) Acid-base Titrations or Neutralisation Titrations: The reaction in
which an acid reacts with a base to give salt and water is called a
neutralisation reaction and the titration involving such a reaction is
called neutralisation titration. An example is the reaction between
NaOH and HCl as shown below.
NaOH + HCl → NaCl + H2O
The indicators used in these titrations, depend upon the pH at the end
point, the familiar examples are phenolphthalein and methyl orange.
22 ii) Oxidation-Reduction or Redox Titrations: Titrations involving

oxidation-reduction reaction, i.e., those in which one component gets
oxidised while the other gets reduced are known as redox titrations. An
example is the titration between oxalic acid and potassium
permanganate in acidic medium. In this case, potassium permanganate
gets reduced to Mn2+ while oxalic acid gets oxidized to CO2 and water. In
this titration, potassium permanganate acts as a self-indicator. The
following equation represents the reaction:
2KMnO4 + 5H2C2O4 + 3H2SO4 → 2MnSO4 + K2SO4 + 8H2O + 10CO2
Chromatometry and iodometry which are discussed in this course are

also redox titrations.
iii) Precipitation Titrations: In certain reactions, when the two components

react, a precipitate is formed. The end point is indicated by the
completion of precipitation. Such reactions are termed as precipitation
reactions and the titrations as the precipitation titrations; an example
is the titration between potassium chloride and silver nitrate as per the
following equation:
KCI + AgNO3 → AgCl + KNO3
Titrations involving AgNO3 are also called argentometric titrations.
iv) Complexometric Titratons: A complexation reaction involves the

replacement of one or more of the co-ordinated solvent molecules, which
are co-ordinated to a central metal ion, M, by some other groups. The
groups getting attached to the central metal ion are known as ligands, L.
M(H2O)n + nL MLn + nH2O
The titration involving such type of a reaction is called a complexometric

titration. For example, you will be using ethylenediaminetetraacetic acid
(EDTA) as the complexing agent in your experiments. The indicator used in
this case is eriochrome black T.
1.6 INSTRUMENTAL DETERMINATION OF

EQUIVALENCE POINT
In HCl vs. NaOH titration described in the last section, determination of the
equivalence point of the titration was detected by colour change of an
indicator. Suppose we don’t want to use an indicator or many a times suitable
indicator may not be available for a titration or the concentration ranges may
be smaller than those required for colour change using an indicator. What
should we do in these situations?
In these cases, instrumental methods which measure some physical property
of the solution are used to detect the equivalence point. These are
conductometer, potentiometer and colorimeter which measure the
conductance, the potential and the colour intensity of the solution,
respectively. Accordingly, these are named as conductometric titrations,
potentiometric titrations and colorimetry. Instrumental methods are quicker
and more accurate. Let us briefly understand these types. 23
Conductometric Titrations
In conductometric titrations, the conductance of the solution being titrated is

measured as a function of the volume of the titrant using a conductometer and
a graph is plotted between the two (Fig. 1.13).
Fig. 1.13: Conductance curve: Titration of NaOH against HCl.
The change in the slope of the conductance vs. volume curve indicates the
equivalence point. The point of intersection of conductance curves for the
titrand having excess of hydrochloric acid (curve I) and excess of sodium
hydroxide (curve II) is the equivalence point.
Potentiometric Titrations
In potentiometric titrations, the solution with unknown concentration is made

the electrolyte in a half cell using an appropriate electrode. The potential of the
half cell with respect to a reference electrode is measured as a function of the
volume of the titrant. The change in slope of the potential vs. volume curve
indicates the end point. A special case of potentiometry where hydrogen ion
concentration, i.e. pH is measured is referred to as pH vs. the volume of the
titrant. The region where a sharp change in pH takes place, in this case pH 7,
indicates the end point (Fig. 1.14).
Fig. 1.14: pH metric titration of HCl vs. NaOH.
Colorimetry
Besides the above two instrumental methods which have been explained as
part of the titrimetric procedures, you will be using yet another instrumental
24 method, viz., colorimetry in your other courses. This is based on the
measurement of the absorption of light of a suitable wavelength by a given
solution. The amount of light absorbed is directly proportional to the
concentration of a given absorbing species. This property is made use of in
determining the concentration of the absorbing solution.
In the next section you will learn about another important aspect of a
Chemistry laboratory that applies to all kind of laboratory work. Before
proceeding further, you would like to assess what you have learnt about the
instrumental determination of equivalence point. So, try answering the SAQ
given below:
SAQ 6
Tick on the correct statement/s.
Instrumental methods are preferred over indicator methods because,
i) very small concentrations of substances can be estimated using

instrumental methods.
ii) instruments are not very expensive.
iii) instrumental methods require a lot of time and do not give accurate
results.
iv) instrumental methods do not require indicators.
1.7 SAFETY MEASURES IN THE LABORATORY

You will be using a number of reagents and chemicals while doing the
experiments. There are laboratory assistants to help you to get these
reagents. Most of these chemicals are kept in the reagent shelves and are
properly labelled. The bench shelves have mostly the liquid reagents which
include hydrochloric, sulphuric and nitric acids. Besides these, other solutions
like silver nitrate, ammonium hydroxide, sodium hydroxide, barium chloride,
etc., may also be kept there. You have to be very careful while using all these,
especially, the acids. Mishandling any chemical may result in injury.
The solid reagents are usually kept on a common table. You should use a
spatula and take only the required amount of the compound from the bottle or
the pack. Don’t waste any chemical. The liquid reagents should be taken with
the help of droppers.
The special chemicals and solutions required for any particular experiment will
be provided by your counsellor at the time of performing the experiment.
An important aspect in a Chemistry laboratory is your own and your fellow

workers safety. Accidents occur in the laboratory because of carelessness and
inadequate knowledge about the chemicals being used. Though accidents
cannot be fully eliminated, yet these can be prevented to some extent by
knowing in advance some general precautionary measures. The following do’s
and don’ts in the laboratory would help you to avoid accidents.
25
The Do’s:
• Wear a laboratory coat or an apron when working in the laboratory.
• Keep the test tubes pointing away from yourself and others while heating
on a burner.
• Use splinters and not a paper to light a burner.
• You should know where the fire extinguishers are located in the laboratory
and how to use them.
• Always use safety goggles for protecting your eyes from a dangerous
operation, e.g., distillation of an inflammable liquid or while doing sodium
ignition test.
• Wash your hands with soap when you leave the laboratory after an
experiment.
• Carry out the reactions involving pungent or noxious fumes under a fume
hood.
• Ensure that gas and water taps are closed before leaving the laboratory.
The Don’ts:
• Don’t wear loose clothes specially the synthetic ones while working in the
laboratory.
• Don’t taste any chemical, not even sucrose; it may be contaminated.
• Don’t pipette out corrosive liquids by sucking with your mouth.
• Don’t put the reagents back into the bottles or packs after use. These
should be poured into another glass bottle kept especially for the waste
liquids.
• Don’t try to insert glass tubing or thermometer into corks forcibly.
• Don’t inhale the vapours of any chemical deeply which might cause
suffocation and choking: be alert and quick in perceiving the smell of the
vapours, keeping the test tube in a slanting position.
• Don’t keep inflammable solvents like petrol, ether, alcohol, etc. near a
burner.
• Don’t add pumice stones to a boiling liquid; add them before beginning to
heat the liquid.
• Don’t ever perform unauthorized experiments and never work alone in the
laboratory.
• Don’t touch electric switches with wet hands.
However, even if you are a careful worker and follow the general rules of
safety, the accidents can occur–that’s why they are called accidents. For such
occasions, you must be fully equipped and must know what to do in such a
26 case. There should be a first-aid box in every laboratory containing some
common things like Dettol, Burnol, Band-aid, bandages, cotton, etc. Generally,
the most common accidents that occur are cuts, burns, fires, poisoning and
rarely, an explosion. A list of hazardous chemicals and their effects is given in
Table 1.2.
Table 1.2: List of hazardous chemicals and their effects
Hazardous Chemicals Effects
Salts of Ag, As, Ba, Cu, Hg, Ni, Pb, Sb, Most of these are very dangerous but only
Ti, V, C2O24− , F , MnO −4 if swallowed, AgNO3 causes caustic burns.
−
H2 S Almost as poisonous as HCN. Exposure

dulls the sense of smell.
SO2, NO2, Cl2, Br2, I2, HNO3, H2SO4, HF All are dangerous as well as unpleasant.
When concentrated, all cause rapid
destruction of the skin; HF is especially
dangerous.
HClO3, HClO4 and their salts Highly oxidising.
Chlorinated alkanes, e.g., CHCl3, CCl4 Most of these are narcotic, causing mental
confusion.
Benzene Toxic vapours causing dizziness
Benzoyl chloride Irritant
Ether, ethanol Very highly inflammable
Nitrobenzene Toxic vapours
Phenol Burns the skin.
Let us know the first-aid that should be given to a student or any person
working in the laboratory, when such a mishap occurs.
i) Cuts: The most common accidents in the chemistry laboratory are cuts
from broken glassware. If you have a cut, wash the wound well with cold
water immediately. If bleeding is severe, apply pressure directly on to the
wound to stop the bleeding. Then an antiseptic cream can be applied to
the wound with a proper dressing.
ii) Burns: Burns generally caused by hot equipment can be treated as the
cuts are treated, that is, wash the burnt part with cold water for sometime
and then apply Burnol to it.
iii) Fire: A small fire in a beaker, caused by the vapours of an inflammable

liquid, can be extinguished by covering it with a watch glass.
If the clothes catch fire one should lie on the floor and fire can be
smothered by wrapping a blanket around the body.
iv) Poisoning: If one happens to swallow a poisonous chemical, plenty of

water should be given if the person is conscious. For a corrosive poison,
calcium hydroxide solution (lime water) should be given as soon as
possible. An antidote should be given only in the case of non-corrosive
poisons. 27
v) Explosion: Sometimes a faulty technique during the experiment can lead
to an explosion. ‘You should work with highly oxidizing or explosive
chemicals only under strict supervision’.
Table 1.3 gives the remedies for a few common chemical reagents used in the
laboratory.
Table 1.3: Remedies for a few chemical reagents
Chemical Neutralising wash
Acid like HNO3, H2SO4, HCl Initial action should be a thorough washing with cold
water. Then NaHCO3 or 2M ammonium carbonate
(leaves no residue on clothes), apply Vaseline or a
soothing cream.
Alkalies, e.g., NaOH, KOH 1M acetic acid, then apply Vaseline or a soothing
etc. cream.
Bromine 2M Ammonia, keep the affected part dipped in

NaHSO3 till bromine is washed off, then apply
Vaseline.
Phenol Ethanol and then hospital treatment
Sodium Ethanol on a cotton wood pad
1.8 ANSWERS
Self-Assessment Questions
1. The pipette is calibrated to include the liquid column trapped at the tip.
Further, blowing it makes it dirty and CO2 in the breath may react with the
solution being pipetted.
2. (5 + 2 + 1) g + (200 + 100 + 50) × 0.001 g + 8 × 0.001 g + 2 × 0.0002 g.
= 8 g + 0.350 g + 0.008 g + 0.0004 g
= 8.3584 g
1000 m
3. From Eq. 1.1, M = mol dm-3
MmV
Where Mm = 40 g mol-1
m = 4.000 g
V = 500 cm3
Therefore,
1000 × 4.000
M= dm −3 = 0.200 mol dm-3
40 × 500
Thus, molar concentration = 0.200 M

28
4. Again consider Eq. 1.1,
1000 m
M= dm − 3
MmV
Where Mm = 169.87 g mol−1
V = 1 dm3 = 1000 cm3
M = 0.1 M
On substituting these values in the above equation, we have
0.1 × 169 .87 × 1000

m=
1000
= 16.987 g
Thus, mass of AgNO3 required for 0.1M solution = 16.987 g.
5. (a) i) NaOH is hygroscopic,
ii) It is not available in pure form as it combines with CO2 from the air
and some part of it is converted into sodium carbonate.
(b) Benzoic acid fits most of the criteria, but its solubility in water is low,
although in non-aqueous solvents such as ethanoic acid (acetic acid)
or ethanol it is not so.
6. i)
ii) ×
iii) ×
iv)
29
EXPERIMENT 1
DETERMINATION OF SODIUM
CARBONATE AND SODIUM
HYDROGEN CARBONATE
PRESENT IN A MIXTURE
Structure
1.1 Introduction 1.5 Observations
Expected Learning Outcomes 1.6 Calculations
1.2 Principle 1.7 Results
1.3 Requirements 1.8 Answers
1.4 Procedure
1.1 INTRODUCTION
You might be familiar with the basic principle of acid-base titrations. In this
experiment we are expanding acid-base titration methods further for the analysis of
a mixture of sodium carbonate (Na2CO3) and sodium hydrogen carbonate
(NaHCO3) or sodium bicarbonate. This method of titration will help you in
understanding the basic principle of some important industrial analyses such as that
of soda ash (anhydrous sodium carbonate), washing soda (hydrated sodium
carbonate, Na2CO3.10H2O), baking soda (sodium hydrogen carbonate), mixture of
sodium carbonate−sodium hydroxide, commercial caustic soda (NaOH), etc. All
these commercial products have a tendency to absorb moisture and carbon
dioxide from atmosphere. This phenomenon of absorption of moisture and carbon
dioxide by these chemicals is called weathering. After such weathering soda ash
and washing soda contain appreciable moisture and sodium hydrogen carbonate
and caustic soda has appreciable amount of moisture and sodium carbonate.
Therefore, it becomes necessary to analyse the purity of these chemicals before
using them for any fine use. The procedures, such as, conductometry,
potentiometry or acid-base indicator methods can be used to analyse the above
substances. Here, we will discuss the acid-base indicator method only for the
30 analysis of a mixture of sodium carbonate and sodium hydrogen carbonate.
Experiment 1 Determination of Sodium Carbonate and sodium Hydrogen Carbonate Present in a Mixture
Procedure used for the titration of a mixture of sodium carbonate and sodium
hydrogen carbonate is basically the same as that of the acid-base titration which
you have performed in your earlier classes, except that there are two analytes in
our sample.

After performing the experiment given, you should be able to:
state and explain the principle of acid-base titration with special reference to
the titration of sodium carbonate and sodium hydrogen carbonate
mixture;
prepare a standard solution of sodium carbonate;
standardise the given solution of hydrochloric acid and use it in estimating the
strength of basic solutions; and
determine the strength of sodium carbonate and sodium hydrogen

carbonate in a given solution.
1.2 PRINCIPLE
During an acid-base titration, the pH changes in a characteristic way. The pH
changes during titration can be understood by plotting a pH curve. A pH curve is
formed, if the pH of the solution being titrated is plotted against the volume of
solution added. The titration of sodium carbonate with a strong acid such as HCl
produces the titration curve shown in Fig. 1.1.This titration curve has two
equivalence points. You may like to ask, why does sodium carbonate solution
behave this way? To answer this question we should study the behaviour of
sodium carbonate in aqueous solution.
Sodium carbonate is a salt of a weak acid and a strong base; when such salts are
dissolved in water, they behave as bases due to the basicity of the conjugate base
CO 32 − . The equilibrium, which is often called hydrolysis, is given by the reaction:
CO 32 − + H2O HCO 3− + OH– ...(1.1)

(carbonateion) (hydrogen carbonate ion)
The hydrogen carbonate ion is furtherhydrolysed to carbonic acid:
HCO 3− + H2O H2CO3 + OH– ...(1.2)

(carbonic acid)
The OH–ions so produced in solution are responsible for the basic character of sodium
carbonate.
When sodium carbonate is titrated with a strong acid, such as hydrochloric acid, the
reaction is completed in two steps. First, the carbonate ions are converted to the
hydrogen carbonate ions, and then to carbonic acid. This is due to the fact that a
strong acid displaces a weak acid from the conjugate base of the latter.
CO 32 − + H+ → HCO 3− ...(1.3)
(Na2CO3) (HCl)
31
HCO 3− + H+ → H2CO3 ... (1.4)

(HCl)
14
1st Equivalence point
pH
2nd Equivalence point
3
Volume of strong acid added in cm
Fig. 1.1: The titration of sodium carbonate with hydrochloric acid.
As neutralisation takes place in two steps (it is indicated by reactions in Eqs. 1.3 and
1.4), we observe two regions of sharp pH change in the titration curve (Fig. 1.1) and
thus two equivalence points at pH 8.31 and pH 3.69. As shown in Fig. 1.1, at the first
equivalence point (in the pH range 9-7) CO 32 − is neutralised to HCO 3− and at the
second equivalence point (in the pH range 5-3) HCO 3− is neutralised to H2CO3.
Combining both the above equations we can write complete neutralisation reaction of
sodium carbonate as
CO 32 − + 2H+ → H2CO3 (1.5)
Finally, the carbonic acid produced as a result of these titrations can

decompose into carbon dioxide (CO2)
H2CO3 (aq) → CO2 (g) + H2O
In this experiment, we will utilise this behaviour of sodium carbonate in the estimation
of a mixture of sodium carbonate and sodium hydrogen carbonate. Let us now
understand the behavior of a mixture of sodium carbonate and sodium
hydrogen carbonate during titration with hydrochloric acid.
The titration curve for a sodium carbonate and hydrogen carbonate mixture is shown
in Fig. 1.2. As you can see, it has two equivalence points. The first equivalence point
indicates half neutralisation of the carbonate in the given sample, i.e., its conversion
to hydrogen carbonate (cf Eq. 1.3). The second equivalence point indicates
neutralisation of the hydrogen carbonate in the initial sample mixture and the
hydrogen carbonate just generated from the half neutralisation of carbonate
(cf Eq. 1.4). In this experiment we will be using acid base indicators to detect
32 both the equivalence points.
14
Na2 CO3 + 1st Equivalence point

NaHCO3
7
+ -
pH 2- H HCO 3-
CO 3 HCO3- HCO3 nd
H
+ 2 Equivalence point
H2CO3
H2CO3
33
VolumeofofHCl
Volume H Cladded
added in
in cm
cm
Fig.1.2:Titration curve for sodium carbonate and sodium hydrogen carbonate titrated
with hydrochloric acid.
As you know, acid-base indicators are organic dyes which change colour as
pH changes. This is because the indicator has two forms, one is acid form in
lower pH medium and other is base form in higher pH medium and having two
different structures. For example, phenolphthalein has two forms, one is
benzenoid form (I) in low pH medium (pH < 8) and thus, it is colourless. Its
second form has quinonoid structure (II) in high pH medium (pH > 10) which
has a pink colour. Similarly, methyl orange exists in quinonoid form (III) in
lower pH medium (pH < 3.2) and benzenoid form (IV) in higher pH medium
(pH > 4.4). The colour of its benzenoid form is yellow while that of quinoniod
form is red.
HO O
OH
-
O
C
O OH-
-
O
H+
O O
Benzenoid form in acidic Quinonoid form in basic
solution (colourless) solution (pink)
(I) (II)
Phenolphthalein
CH3
-
O 3S N N N
CH3
Benzenoid form in basic solution (yellow)
(III)
+
OH- H
CH3
- +
O 3S NH N N
CH3
Quinonoid form in acidic solution (red)
(IV)
Methyl orange
33
Equivalence point so obtained using indicators is called an end point. It is not

necessary that the end point is coincident with the equivalence point, because
The pH range is termed of the delay in getting the indicator to show the colour change, and other
as the colour-change
factors. Ideally end point and equivalence point should be as close as
interval of the pH
indicator. The position of
possible.
the colour-change
In general acid-base indicators show colour change in a pH range of ±1 pH
interval in the pH scale
unit. The colour change and the pH range of some common indicators are
varies widely with
different indicators. For tabulated below to guide you for selecting appropriate indicator for any acid
most acid-base titrations, base titration.
we can, therefore, select
an indicator which Table 1.1: Colour changes and pH ranges of acid-base indicators
exhibits a distinct colour
Name Acid Colour pH Range of Base Colour
change at a pH close to
the equivalence point. Colour Change
For example, pH
Methyl violet Yellow 0.0 - 1.6 Blue
indicator phenolphthalein
shows colour change in Thymol blue Red 1.2 - 2.8 Yellow
pH range 8-10 and
methyl orange in the pH Methyl orange Red 3.2 - 4.4 Yellow
range 3.1-4.4. Therefore,
for detection of end Bromocresol Yellow 3.8 - 5.4 Blue
points in the pH range of green
8-10, phenolphthalein
will be the suitable Methyl red Red 4.8 - 6.0 Yellow
indicator. Similarly for
Litmus Red 5.0 - 8.0 Blue
detection of the end point
in pH range of 3-4, Bromothymol Yellow 6.0 - 7.6 Blue
methyl orange will be the
blue
suitable indicator.
Thymol blue Yellow 8.0 - 9.6 Blue
Phenolphthalein Colourless 8.2 - 10.0 Pink
Using Fig. 1.2 and Table 1.3, you can select phenolphthalein and methyl orange as
most suitable indicators for the detection of the first and the second end points,
respectively for the titration of mixture of sodium carbonate and sodium hydrogen
carbonate. Once, these two end points are detected, the volume of HCl used to titrate
sodium carbonate and sodium hydrogen carbonate in the mixture can be determined.
This can be further illustrated by considering Fig. 1.3.
−
CO 2
−
HCO 3 HCO 3− H2CO 3
(Total hydrogen carbonate)
(V1 = Vb - Va)
(V2 = Vc - Va)
2V1
(V3=V2-2V1)
Va initial Vb (1st end point) Vc (2nd end point)
Fig. 1.3: Volumes of HCl used during titration of a mixture of sodium carbonate
34 and sodium hydrogen carbonate.
In this diagram Va, Vb and Vc refer to burette readings−initial, at the 1st end point
with phenolphthalein and at the 2nd end point with methyl orange, respectively.
These values of Va, Vb and Vc are also used in calculating the volumes of HCl
required for neutralizing Na 2CO 3 and NaHCO3 present in mixture. Thus, for the first
end point we need Vb –Va = V1 cm3 and for the second end point Vc–Va= V2 cm3 of
hydrochloric acid. Thus, V1 is the volume of HCl needed to titrate half of the
+
sodium carbonate initially present in the mixture (CO 32 − + H → HCO 3− ) and 2V1
will be the volume of HCl needed to neutralise whole sodium carbonate in the
+
mixture (CO 32 − + 2H → H2CO3). V2 is the volume of HCl required to neutralize
both sodium carbonate and sodium hydrogen carbonate in the given solution. On
subtraction of 2V1 from this volume, V2, we can find out the volume of HCl, V3,
needed to neutralise initially present sodium hydrogen carbonate in the
mixture,V3 = (V2 – 2V1).
The corresponding chemical reactions may be summarised as:
2− + − end point with phenolphthalein,

CO 3 + H HCO 3
Volume of HCl = V1 = Vb − Va
(Na2CO3) (HCl)
+
HCO 3 +
− H H2CO 3
(NaHCO3 (HCl) end point with methyl orange,
− Volume of HCl = V2 = Vc − Va
+ HCO3 of
sodium
carbonate)
Using volume V1, V2, V3 and molarity equations we can calculate amount of sodium
carbonate and sodium hydrogen carbonate in the mixture.
Using reaction of Eq. 1.3, we can write molarity equation for the half
neutralisation of sodium carbonate, here sodium carbonate reacts with
hydrochloric acid in 1:1 molar ratio to gives hydrogen carbonate. Thus the
molarity equation for this step will be:
M Na 2CO3 VNa2CO3 1
=
M HCl VHCl 1
i.e. MNa 2CO3 VNa 2CO3 = MHCl VHCl (1.6)
Where MNa2CO3 the molarity of sodium is carbonate solution and VNa 2CO3 is the
volume of sodium carbonate used in titration. M HCl is the molarity of
hydrochloric acid and VHCl is the volume of the hydrochloric acid used in the
titration of sodium carbonate up to hydrogen carbonate stage.
As indicated by the final reaction (Eq. 1.5), complete sodium carbonate reacts with
hydrochloric acid in 1:2 molar ratios. Hence, molarity equation can be written as
MNa 2 CO 3 VNa 2 CO 3 1
=
MHCl VHCl 2
35
i.e. 2 MNa 2CO3 VNa 2CO3 = MHCl VHCl (1.7)
Where MNa2CO3 is the molarity of sodium carbonate solution and VNa 2CO3 is the
volume of sodium carbonate used in titration. M HCl is the molarity of
hydrochloric acid and VHCl is the volume of the hydrochloric acid used in the
complete titration of sodium carbonate (i.e. volume of HCl used up to methyl
orange end point for standard sodium carbonate solution)). Eq. 1.7 will be
used for the calculation of the strength of hydrochloric acid in the titration of
standardisation of hydrochloric acid.
Chemical reaction of the neutralisation of sodium hydrogen carbonate with

HCl can be written as:
HCO 3− + H+ → H2CO3 ...(1.8)
Here sodium hydrogen carbonate reacts with hydrochloric acid in 1:1 molar ratios.
Hence, molarity equation can be written as
M NaHCO 3 V NaHCO 3 = MHClVHcl ...(1.9)
Where MNaHCO3 is the molarity of sodium hydrogen carbonate solution and VNaHCO 3
is the volume of sodium hydrogen carbonate used in titration. M HCl is the
molarity of hydrochloric acid and VHCl is the volume of the hydrochloric acid
used in the titration.
Hydrochloric acid used for titration of the mixture solution is not a primary standard.
Therefore, before using hydrochloric acid for the titration, it should be standardised
with a suitable primary standard, preferably sodium carbonate. The reaction
between sodium carbonate and hydrochloric acid is shown by Eq .1.5 and molarity
equation used for calculating the molarity of hydrochloric acid is shown by Eq. 1.7.
End point of the titration is detected with methyl orange indicator.
Before proceeding further, answer the following SAQs.
SAQ 1
Suggest whether aqueous solutions of the following substances are acidic, basic or
neutral.
a) NaCN; b) NaCl; c)CH3COONa; d) NaHCO3; e) K2CO3
SAQ 2
Predict the number of pH breaks or sharp pH change(s) which will be observed for
the following titrations:
a) CH3COOH –NaOH
b) NaHCO3 –HCl
c) K2CO3 – HCI
36
SAQ 3
On the basis of Fig. 1.4 given below, suggest suitable indicators for the titration of
ethanedioic acid or oxalic acid (H2C2O4) against strong base.
14
2-
C2 O4
nd
2 Equivalence point
-
PH 7 HC2 O4
1st Equivalence point
H2 C2 O4
3
Volume of strong base added in cm
Fig. 1.4: Titration of ethanedioic acid (oxalic acid) with strong base like NaOH.
1.3 REQUIREMENTS
You will need the following apparatus and chemicals for this experiment.
Apparatus Chemicals
3
Beaker (250 cm ) 2 Hydrochloric acid
3
Burette (50 cm ) 1 Methyl orange
Burette stand with clamp 1 Phenolphthalein
3
Conical flask (250 cm ) 1 Sample: Mixture of sodium carbonate and
sodium hydrogen carbonate or baking
soda
Funnel 1 Sodium carbonate (AR grade)
3
Pipette (20 cm ) 1
3
Volumetric flasks (250 cm ) 2
Weighing bottle 1
Solutions provided
Sample solution: Prepare a sample solution by dissolving a mixture of

sodium carbonate and sodium hydrogen carbonate (8.5 g Na2CO3+5.4 g
NaHCO3) or commercial baking soda (15 g) in 2 dm3 distilled water. 37
Phenolphthalein indicator solution: It is prepared by dissolving 0.1 g of

the reagent in 80 cm3 of ethanol and adding adequate distilled water to
make it 100 cm3. If a precipitate is formed, it is filtered.
Methyl orange indicator solution: It is prepared by dissolving 0.1 g of free

acid/sodium salt of the indicator in 80 cm3 of distilled water and adds 20 cm of
ethanol to make it 100 cm3.
Hydrochloric acid solution (~ 0.1 M): This solution is prepared by taking 8.5
cm3 conc. HCI (37%) in a 1 dm3 volumetric flask and diluting the acid up to the
mark with distilled water.
1.4 PROCEDURE
To obtain satisfactory First collect 0.1 M hydrochloric acid in a 250 cm3 beaker. Since hydrochloric
results by double indicator acid is a secondary standard, you have to standardise it by titrating it against a
method the solution primary standard, Na2CO3 in this case.
titrated must be cold, and
loss of carbon dioxide 1) Standardisation of hydrochloric acid:
must be prevented as far
as possible by keeping i) Take exact mass of a clean dry weighing bottle and then weigh the
the tip of the burette weighing bottle with about 1.35 - 1.40 g of dried sodium carbonate
immersed in the liquid exactly. Transfer the sodium carbonate to a clean volumetric flask of
during titration. 250 cm3 capacity through a glass funnel. Find the exact mass of
sodium carbonate transferred by subtracting the mass of empty
weighing bottle from the mass of the weighing bottle plus sodium
Before using burette and carbonate. Dissolve sodium carbonate in volumetric flask in distilled
pipette you should know water and make up the volume to the mark with distilled water.
how to use them.
ii) Fill up the burette with hydrochloric acid solution and mount it on a
stand. Note the reading on the burette and record it in the
Observation Table I under the initial reading column.
Your apparatus should
by dry and clean. iii) Pipette out 20 cm3 of the standard sodium carbonate solution, add
two to three drops of methyl orange indicator. Titrate with constant
swirling against a white background till a redcolour is obtained.
Record your reading in the Observation Table I under the final
reading column. Repeat the titration to get at least two concordant
readings.
2) Titration of the sample solution (mixture of sodium carbonate and

sodium hydrogen carbonate) against standardised hydrochloric
acid:
i) Now collect sample solution in a 250 cm3 beaker. Pipette out 20 cm3
of the sample solution in a conical flask. Add 1-2 drops of
phenolphthalein indicator to it; a pink colour will be obtained.
ii) Note the initial reading of the burette in the Observation Table II
under the initial reading column. Run in a standardised HCI from the
burette slowly into flask until the pink colour is just discharged. Note
the burette reading in the Observation Table II under the 'reading with
38 phenolphthalein' column.
iii) Now, add a few drops of methyl orange to the solution in the conical
flask; a yellow colour is obtained. Run in a further quantity of the acid
until the yellow colour of the solution changes to red. Note the final
burette reading in the observation Table II under the 'reading with
methyl orange' column. Repeat the titration with both the indicators to
get two concordant sets of readings.
1.5 OBSERVATIONS
Mass of the weighing bottle ................. = m1 g = ..g
Mass of weighing bottle + sodium carbonate . = m2 g =. ..g
Amount of sodium carbonate transferred ...... = m2− m1 = mg = ..g
Molar mass (Mm) of sodium carbonate = 106 g mol–1
Volume of sodium carbonate prepared = 250 cm3
Molarity of sodium carbonate solution = M Na 2 CO 3 = ?
m × 1000
MNaCO 3 = mol dm − 3
Mm × 250
m× 4
= mol dm −3
106
M NaCO 3 = .......... mol dm −3
Observation Table I
Sodium carbonate vs. Hydrochloric acid
3
S. No. Volume of Burette reading Volume of HCl in cm
3
Na2CO3 in cm (Final – Initial)
(V Na 2 CO 3 ) Initial final (VHCl)
Observation Table II
Sample solution (Mixture of Na2CO3 and NaHCO3) vs. Hydrochloric acid
S. Volume Burette reading Volume of HCl Volume of HCl Volume of HCl

No. of used in titration used in used in
sample of half of titration of titration of
Initial With With
solution Na2CO3 total initially
3 phenolphthalein methyl
in cm 2−
(CO 3 →HCO 3 )
− 2−
CO 3 + HCO 3
− present
orange
3 3 NaHCO3
in cm in cm in cm
3
Vs Va Vb Vc (V1=Vb–Va) (V2=Vc–Va) (V3=V2–2V1)
39
1.6 CALCULATIONS
a) Standardisation of hydrochloric acid solution:
Molarity of sodium carbonate solution = MNa2CO3 = ..............mol dm-3
Volume of sodium carbonate solution = V Na 2 CO 3 = 20cm3
Volume of hydrochloric acid = VHCl = .......cm3
(from Observation Table I)
Molarity of HCl solution = MHCl = Mstd= ?
Using Eq. 1.7, Molarity of HCl solution,
2 M
Na 2 CO 3VNa 2 CO 3
M std = = .............. mol dm-3 ...(1.10)
VHCl
b) Estimation of sodium carbonate and sodium hydrogen carbonate

in the sample solution: This can be done as follows:
i) Estimation of sodium carbonate in the Sample Solution:

The molarity of the Volume of hydrochloric acidusedin the titration of half of sodium
sodium carbonate in the carbonate in the given sample (from Observation Table II)
given sample can also
be calculated using V1 = ......... cm3
molarity Eq. 1.10. But in
this case the volume of Molarity of hydrochloric acid solution MHCl = Mstd =.......... mol dm-3
HCl used in titration will
be 2V1. (From standardisation of HCI solution, Eq. 1.10)
Volume of the sample solution Vs cm3 = 20cm3
Molarity of Na2CO3 in the given sample solution = Msc = ?
Using Eq. 1.6, substituting values from above:
MscVs = MstdV1
Mstd V1
Msc =
Vs
=...........mol dm-3
Strength of sodium carbonate present in the
given sample solution = Msc× Molar mass of sodium carbonate
=......... g dm-3
ii) Estimation of sodium hydrogen carbonate in the Sample

Solution:
Volume of hydrochloric acid used in the titration of sodium

hydrogen carbonate in the given sample
40 (from Observation Table II) = V3 = .......cm3

Molarity of hydrochloric acid solution = MHCl = Mstd = .......mol dm-3
(From standardisation of HCI solution, Eq. 1.10)
Volume of the sample solution Vs = 20 cm3
Molarity of sodium hydrogen carbonate in the given sample

solution = Mshc= ?
Similar to Eq. 1.8 neutralisation reaction of sodium hydrogen

carbonate and HCl can be written as:
HCO 3− + H+ → H2CO3
Here sodium hydrogen carbonate reacts with hydrochloric acid in 1:1

molar ratio. Hence, molarity equation can be written as
M HCO− V HCO− = MHClVHcl

3 3
Therefore, on substitution of the values from above:
Mstd V3
Mshc =
Vs
=............. mol dm-3
Strength of sodium hydrogen carbonate in the given Sample

Solution = Msch × Molar mass of sodium hydrogen carbonate
(Molar mass of NaHCO3 = 84 g mol–1)
= .......... g dm-3
1.7 RESULTS
i) Molarity of Na2CO3 in the given solution = ..............mol dm-3
Molarity of NaHCO3 in the given solution = ..............mol dm-3
ii) Strength of Na2CO3 in the given solution = ..............g dm-3
Strength' of NaHCO3 in the given solution = ..............g dm-3
Compare the results with the correct values for the given sample solution of
the mixture of sodium carbonate and sodium hydrogen carbonate.
Using the experimental technique mentioned above, you can also design an Sodium hydroxide and
experiment to determine the percentage purity of commercial soda ash, commercial caustic soda
washing soda, etc. For analysing mixture of sodium carbonate and sodium absorb moisture and CO2
hydroxide, and commercial caustic soda, there is need of modification in the from the air and get
calculation of the volume of HCl needed to neutralise them. In this analysis, converted into
carbonate.
NaOH and half of the Na2CO3 is neutralised at the phenolphthalein end point
and remaining half of sodium carbonate i.e. hydrogen carbonate at methyl
orange end point. Calculation is carried out using following equations: 2NaOH + CO2 →
V
V V
Na2CO3 + H2O
M
2( − phth )
2
mo HCl
Na2CO3 =
s
41
M
M
[ − 2( − phth )]
V
phth mo HCl
NaOH =
s
Where, M Na 2 CO3 and M NaOH are the molarities of sodium carbonate and sodium
hydroxide in the sample, respectively. Vs is the volume of the sample used in
titration. Vphth and Vmo are the volumes used in titration of HCl up to
phenolphthalein end point and methyl orange end point, respectively. MHCl is
the strength of standardized HCl used in this titration.
1.8 ANSWERS
1. a) Basic
b) Neutral
c) Basic
d) Basic
e) Basic
2. a) One
b) One
c) Two
3. Methyl orange for the first pH break and phenolphthalein for second pH
break (See Fig 1.4). As methyl orange remains coloured after the first end,
point therefore, pH meter is preferred.
42
Experiment 2 Estimation of Oxalic Acid by Redox Titration
EXPERIMENT 2
ESTIMATION OF OXALIC ACID
BY REDOX TITRATION
Structure
2.1 Introduction Principle
Expected Learning Outcomes Requirements
2.2 Oxidation-Reduction: Redox Procedure

Potential
Observations
2.3 Redox Titrations
Calculations
Redox Titration Curves
Result
Redox Indicators
2.5 Answers
2.4 Permanganatometric
Estimation of Oxalic Acid
2.1 INTRODUCTION
You have learnt about the theory and applications of acid-base titrations in
Experiment 1. A large number of analytical determinations make use of another
important kind of titrations, namely, redox titrations. As the name suggests these
titrations are based on oxidation-reduction reactions. In contrast to acid-base
titrations in which the titration reaction involves the formation of undissociated
molecules of a weak electrolyte (water or a weak acid), a redox titration
reaction is associated with the transfer of electrons. The electrons are
transferred from a reducing agent to an oxidising agent. In this experiment we
will make use of such titrations. An attempt is made here to make you
understand some fundamental concepts related to redox titrations and the
theory behind the reactions involved. Analytical redox titrations involve the use
of a variety of oxidising and reducing agents. Different types of redox titrations
are named on the basis of oxidising/reducing agents involved. Of these, two
important types of titrations are permanganatometry using potassium
permanganate, KMnO4 and chromatometry using potassium dichromate,
K2Cr2O7 as the oxidizing agents. In the present experiment you will estimate
oxalic acid using permanganatometry and in Experiment 3 the same oxidising 43
agent would be used to estimate water of crystallization in the Mohr’s salt.

Experiment 4 deals with the estimation of iron using another oxidising agent i.e.
potassium dichromate.

After performing the experiment, you should be able to:
define oxidation, reduction and redox titrations,

explain the significance of redox potential in redox titrations,
interpret redox titration curves,
explain the use of various types of redox indicators,
state and explain the principle involved in permanganatometry, and
apply the redox titration method, viz., permanganatometry for
estimating oxalic acid in the given solution.
2.2 OXIDATION-REDUCTION: REDOX

POTENTIAL
Let us first briefly review the theory of oxidation and reduction reactions.
Oxidation is the process which results in the loss of one or more electrons by an
atom or an ion, e.g.,
Fe2+ Fe3+ + e− ... (2.1)

An oxidising agent is the
one which gains Reduction, on the other hand, is the process which results in the gain of one or
electrons and gets more electrons by an atom or an ion, e.g.,
reduced, e.g., Cl2 in Eq.
2.2 and a reducing agent Cl2 + 2e− 2Cl− (2.2)
is the one which loses
electrons and gets Different oxidising/reducing agents differ from one another in their strength. An
2+
oxidised, e.g., Fe in oxidising agent can behave as a reducing agent in the presence of a stronger
Eq. 2.1. oxidising agent. For a reaction of the type:
Oxidising agent (I) + Reducing agent (II) → Reducing agent (I) + Oxidising agent (II)
where we have a pair of oxidising-reducing agents such that either of the species
can act as an oxidising or a reducing agent, the direction of the reaction is
determined by comparing the redox potentials of the oxidising/reducing
agents. The redox potential is a quantitative characteristics of the oxidising/
reducing power of a reagent. Let us try to understand the significance of redox
potential.
Redox Potential
In a system containing both an oxidising agent and its reduction product, there will
be an equilibrium between them and the electrons. If an inert electrode, such as
platinum, is placed in such a redox system, e.g., one containing Fe3+ and Fe2+ ions,
44
it will assume a definite potential indicative of the position of the equilibrium. If the
3+ 2+
system tends to act as an oxidising agent, Fe → Fe ; it will take electrons from
the platinum electrode leaving the latter positively charged. On the other hand, if the
2+ 3+
system has reducing properties (Fe → Fe ), electrons will be given to the metal
which will then acquire a negative charge. The magnitude of the potential will thus The potential of the
standard hydrogen
be a measure of the oxidising or reducing properties of the system. It is quite
electrode is
difficult to measure this potential between the metal and the solution or between conventionally taken as
different oxidation states of a metal. For this purpose, such a system is coupled with zero, just as the
a Standard Hydrogen Electrode (SHE), i.e., we make a galvanic cell and measure temperature of melting
electromotive force (e.m.f.) of the cell. The e.m.f. of such a cell is the difference of ice on Celcius scale is
potential of the given system and SHE, the potential of which is taken to be taken as zero.
0.00 V. This e.m.f. is referred to as the redox potential.
It should be noted that a solution of a pure oxidising agent or a pure reducing agent
always contains the products of their reduction or oxidation, respectively. For
example, reductant Fe2+ always contains some Fe3+ and the oxidant MnO −4 always
contains Mn2+ ions. That is why it is more correct to speak of the redox potentials of
oxidation-reduction couples such as Fe3+/ Fe2+, Mn72 + / M n2 + , etc., rather than of the
individual oxidant or reductant potentials.
For a simple reduction reaction,
Oxidised form + ne− Reduced form
the reduction potential, E, is given by Nernst Equation:
(2.3)
where R ═ Gas constant (═ 8.314 J mol-1 K-1)
T ═ Absolute temperature (K)
F ═ Faraday’s constant (═ 96,500 C)
Eo ═ Standard redox potential
n ═ number of electrons gained or lost
When [Ox] ═ [Red], the log term in Eq. 2.3 becomes zero, then the potential of
the system is called standard potential. It is a characteristic of a given
system. At 300 K, substituting the values of R, T and F in Eq. 2.3, we get the
following.
(2.4)
Thus, knowing the chemical reaction involved and the potential of the solution,
we can use the Nernst equation to evaluate the relative concentrations of
oxidised and reduced forms. The solution’s potential can also be calculated, if
we know the concentrations of the two forms.
45
SAQ 1
Write equations for the following half reactions:
i) Oxidation of hydrogen molecule to form hydrogen ions
ii) Oxidation of sulphide ions to form sulphur
iii) Reduction of chlorine molecule to form chloride ions
iv) Oxidation of cuprous ions to form cupric ions
v) Reduction of oxygen molecules to form oxide ions
2.3 REDOX TITRATIONS

You know, a redox titration is based upon the oxidation-reduction reaction
between a titrand and a titrant. Here the end point can be detected either by
the colour change of a redox indicator or by plotting data taken by using a
potentiometer. In this section we first discuss the redox titration curves and
then the redox indicators. These concepts will tell you how redox indicator and
potentiometric detection procedures work.
2.3.1 Redox Titration Curves

We can see from Eq. 2.4 that the potential of a given reaction depends upon
the relative concentrations of oxidised/reduced forms. In the course of a redox
titration, the solution potential also changes, since the concentration of oxidised
and reduced forms goes on changing. At one stage, when either of the forms
gets exhausted, i.e., at the end point, there is a sharp change in otherwise
gradually varying potential. You may recall here what you studied in acid-base
titrations, where either the pH or the conductance shows a sharp change at the
end point. We shall theoretically try to see the variation of potential during the
course of titration which is called redox titration curve. Let us illustrate this by
taking an example of the titration between ferrous ions and potassium
permanganate solution.
Redox Titration Curve for Ferrous Sulphate-Potassium

Permanganate Titration
In the titration of FeSO4 with KMnO4 in the acidic medium, the permanganate
ions oxidise ferrous ions to ferric ions and get reduced to divalent manganese
ions. The ionic reactions involved are as follows:
[Fe2 + Fe3 + + e− ] × 5 (2.5)
MnO−4 + 8H+ + 5e− Mn2+ + 4 H2O (2.6)
Adding Eq. 2.5 and Eq. 2.6,
MnO4− + 5 Fe2 + + 8H+ Mn2 + + 5 Fe3 + + 4 H2O (2.7)

2+ 3+
The system contains two redox couples, viz., Fe / Fe and MnO−4 / Mn2+ .
46 Since, both the reactions are in equilibrium, at any stage of titration, the solution
contains all the species. To calculate the potential of the solution theoretically
at any instance of titration, we can make use of Nernst equation using the
standard redox potentials for the two couples; and substituting the values of R,
T and n in Eq. 2.3. The values of n can be obtained from Eq. 2.5 and Eq. 2.6.
Thus:
0.059 [5 Fe3 + ]
E = 0.77 + log (2.8)
1 [5 Fe2 + ]
0.059 [MnO4− ] [H+ ]8

E = 1.51 + log (2.9)
1 [Mn2+ ]
However, it is simpler to use Fe2+/Fe3+ couple in the region before the

equivalence point and MnO−4 / Mn2+ couple after it. This is because it is easier to
calculate the amounts of the corresponding ions under such conditions.
Before we start the titration, we have only ferrous ions in solution. When we
add KMnO4, MnO −4 ions oxidise some of the ferrous ions to ferric ions and a
2+ 3+
potential is developed, point A in Fig. 2.1, between Fe and Fe ions. As we
3+
go on adding more and more of permanganate, the amount of Fe ions goes
2+
on increasing and that of Fe goes on decreasing whereby the potential
(cf. Eq. 2.8) also goes on increasing gradually. At a stage just before the
equivalence point, say about 0.1 cm3 less than that required for the end point,
almost all the ferrous ions are oxidised and the potential is approximately equal
2+ 3+
to the maximum for Fe /Fe system under given conditions, point B in
Fig. 2.1.
Fig. 2.1: Redox titration curve for FeSO4 – KMnO4 titration.
At equivalence point, point C in Fig. 2.1, the potential is given by the following
general equation:
aEa0 + bEb0
E= (2.10)
a+b
where Ea0 and Eb0 are the standard potentials of the reducing and the
oxidising agent, respectively and a and b are the corresponding stoichiometric
coefficients. In the present case Ea0 = 0.77V, Eb0 = 1.51V, a=1, b = 5 and
E =1.387V.
Immediately after the equivalence point the amount of Fe2+ is negligibly small
and is difficult to calculate. The potential can be calculated by making use of
MnO −4 / Mn2+ couple as, at this stage it becomes easier to evaluate the amount 47
of MnO −4 and Mn2+ ions. The potential now corresponds to the minimum for the
MnO −4 / Mn2+ couple, point D in Fig. 2.1. Beyond this, further addition of MnO −4
merely alters the relative amounts of MnO −4 and Mn2+ and there is a gradual
variation in the potential of the solution. The calculated redox potential during
the titration of 100 cm3 of FeSO4 solution at [H+] ═ 1M with permanganate
solution of the same molarity is given in Table 2.1. You would notice that
Eq. 2.9 for determining the potential of MnO −4 / Mn2+ couple contains [H+] term.
The concentration of [H+] is kept 1M so that in the effective equation we need
only the amounts of MnO −4 and Mn2+ ions. However, it may be mentioned
here that the hydrogen ion concentration has an enormous effect upon the
oxidation potential of the oxidising agent, MnO −4 in this case. At a pH of 6,
e.g., it is found that the oxidation potential of permanganate is about 0.6 volt
lower than with 1M acid solution, where pH ═ 0. Use is made of this fact, e.g.,
in the fractional oxidation of halides to the corresponding halogen. At a pH of 5
or 6, iodide is oxidised to I2 by permanganate, whereas bromide and chloride
are not affected. At a pH of about 3 (acetic add), bromide is oxidised, but
chloride is still unaffected. The latter is oxidised only at a much higher
hydrogen ion concentration.
Table 2.1: Variations of the redox potential during titration of 100 cm3 of
0.1M FeSO4 solution with 0.02M KMnO4 solution at [H+] ═ 1M
3 3+
KMnO4 Excess cm [Fe ] [ MnO−4 ] Calculation Oxidation
added
2+
[Fe ] E= potential
FeSO4 KMnO4 [Mn ]
2+
3
(cm ) E ( V)
50 50 ─ 50:50=1 ─ 0.77+0.059 log1 0.770
91 9 ─ 91:9≈10 ─ 0.77+0.059 log10 0.829
99 1 ─ 99:1≈100 ─ 0.77+0.059 log100 0.888
99.9 0.1 ─ 99.9:0.1 ─ 0.77+0.059 log 1000 0.947

≈1000
*
100 0.0 0.0 100:0 ─ 0.77 + 5 × 1.51 1.387
(equiv. pt.) 5.1
100.1 ─ 0.1 ─ 0.1:100 0..59 1.475
1.51 + log 0.001
=0.001 5
101.0 ─ 1.0 ─ 1:100 0..059 1.487
1.51 + log 0.001
=0.01 5
110.0 ─ 10 ─ 10:100 0..059 1.498
1.51+ log 0.001
=0.1 5
150 ─ 50 ─ 50:100 0..059 1.507
1.51 + log 0.5
=0.5 5
* These figures show that the equivalence point is not in the middle of the break, as was the case in
titration curves by the neutralisation method.
Redox potential of a redox system remains unaffected by dilution, this is

because dilution affects both the oxidised and the reduced species equally. This
48 is justified because in the Nenst equation also, the relative concentrations and
not the absolute concentrations of the two forms are required. On the other
hand, in the region beyond the equivalence point, the actual potential would be
slightly different from the calculated one, since [H+] ions are involved in the
calculations and their concentration does depend upon dilution. This error,
however, does not affect the general conclusions and can, therefore, be
neglected.
The above description of redox titration curves is given here to make you aware
of the changes in potential taking place during the reaction. Since, you are going
to use only the indicator method to detect the end point, you would not be
required to draw such curves. However, these titrations can also be followed by
actually measuring the potential of the solution with the help of a potentiometer
(pH meter). In such cases, one has to plot these curves and determine the
equivalence point by using derivative curves as was done in case of acid-base
titrations.
2.3.2 Redox Indicators

You have seen how the potential varies during the titration and also that at the Redox indicators are
equivalence point there is a sharp change in the potential. As mentioned before, substances which can be
this change is similar to the sharp change observed in the pH during acid-base reversibly oxidised or
titrations. As you used an acid-base indicator which changes colour in the pH reduced and have
range corresponding to the sharp change in pH at the end point, here we need different colours in
oxidised and reduced
a chemical species which can change colour in the potential range
forms.
corresponding to the sharp change at the end point. A chemical substance
which changes colour when the potential of the solution reaches a definite value
is termed an oxidation-reduction or redox indicator.
In redox titrations, indicators are used in three different ways. These have
already been discussed briefly in the introductory Unit 1. Let us recall it here. In
certain titrations, e.g., those involving KMnO4, one of the reacting species itself
changes colour at the equivalence point and is called a self indicator. In some
cases the indicator needs to be added to the solution, as phenolphthalein or
methyl orange in the case of acid-base titrations, or, diphenylamine in case of
chromatometry. Such indicators are called internal indicators. In yet some
other cases, redox indicator may be replaced by a reagent which is used as a
spot test reagent for the ion being determined. Such indicators are not added
into the solution but are used externally. At various stages of the titration, a
drop of the reaction mixture is taken out and tested for the ion by mixing with
the indicator on a porcelain plate. Potassium ferricyanide, K3 [Fe(CN) 6], is an
example of an external indicator which is used in the titration of Fe2+ ions with
dichromate. Having learnt about oxidation/reduction reactions, redox potential and
redox indicators, you will perform the following experiment based on the
principle of redox titrations.
2.4 PERMANGANATOMETRIC ESTIMATION OF

OXALIC ACID
As stated above, potassium permanganate is a good oxidising agent. It is used in a
number of titrimetric determinations where the method is known as
permanganatometry. One of the most important determinations by this method is 49
titrimetric determination of oxalic acid solution. Let us learn the principle of this
determination.
2.4.1 Principle
Potassium permanganate is an oxidising agent and gets reduced in the presence of
a suitable reducing agent, for example, oxalate ions ( C 2 O 24 − ) in the present case. Its
The oxidizing power of reduction can be brought about in acidic, neutral or alkaline medium. The
permanganate ion is permanganate ion MnO −4 gets reduced to Mn2+ ion in acidic medium as shown in
medium dependent; it is
Eq. 2.6, and to Mn4+ in neutral and alkaline media, as shown in Eq. 2.11.
related to the change in
the oxidation state of
M nO −4 + 8H+ + 5e − Mn2 + + 4H2O, E 0 = 1.51V (2.6)
manganese in a
particular medium. + 7 state +2 state
MnO 4− + 2H2O + 3e − MnO 2 + 4OH− , E 0 = 0.57 V ...(2.11)

+7 state +4 state
Usually titrations involving potassium permanganate are carried out in acidic

medium. This is due to the higher oxidising power of permanganate ion in acidic
medium than in neutral or alkaline medium; secondly, the formation of brown
coloured MnO2 in alkaline medium interferes with the detection of the end point.
While permanganate ion gets reduced, C2O42− gets oxidised to carbon dioxide
as per the following equation
C2O 2 −
4 → 2CO 2 + 2e
− ...(2.12)
The overall ionic equation for the titration of oxalic acid and permanganate in acidic
medium can be obtained by adding Eq. 2.12 and Eq. 2.6 after balancing the
number of electrons between them as follows:
[C2O 2 − −
4 → 2CO 2 + 2e ] × 5 (2.12)
[MnO − + − 2 + + 4H O] (2.6)
4 + 8H + 5e → Mn 2
2MnO 4− + 5C 2O 24 - + 16H+ → 2Mn 2 + + 8H2O + 10CO 2 (2.13)
We see from Eq. 2.13, that two moles of permanganate ions react with 5 moles of
oxalate ions. Therefore, substituting the values in the molarity equation, the
molarities are related as per the following equation:
MKMnO 4VKMnO 4 2
= ⇒ 5MKMnO 4VKMnO 4 = 2VH2C 2O 4 MH2C 2O 4 (2.14)
MH2C 2O 4VH2C 2O 4 5
A slight excess of KMnO4 at the end point imparts a distinct pink colour to the
solution and therefore, acts as a self indicator. You can use Eq. 2.14 as the
The Mohr's salt is preferred molarity equation to find out the molarity of the oxalic acid.
to ferrous sulphate
The solution of KMnO4 is not stable and its strength changes on storage. It is,
because it has better
shelf-life. therefore, a secondary standard. You have to standardise it by titrating against a
suitable primary standard. A number of primary standards can be used for this
purpose. Here you would be using a standard solution of ferrous ammonium
50
sulphate, FeSO4.(NH4)2SO4.6H2O or Mohr’s salt for standardisation of potassium
permanganate. The equation for the titration is given by Eq. 2.7.
[MnO 4− + 8H+ + 5Fe 2 + → Mn2 + + 5Fe3 + 4H2O]
The corresponding molarity equation for the titration would be:
5 MKVK ═ MFVF
Permanganatometry finds its use also in the estimation of hydrogen peroxide,

nitrites and persulphates, etc.
SAQ 2
Write the chemical reaction involving a titration of KMnO4 and ferrous sulphate
solution in presence of dilute H2SO4.
[Hint: Two steps are involved in the reaction]
SAQ 3
Why KMnO4 cannot be taken as a primary standard?
2.4.2 Requirements
Apparatus Chemicals
Burette (50 cm3) 1 Ferrous ammonium sulphate, FAS

FeSO4. (NH4)2 SO4.6H2O,
also known as Mohr’s salt
Pipette (20 cm3) 1 Sulphuric acid (1 M)
Conical flasks (250 cm3) 2 Oxalic acid
Test Tube 1 Potassium permanganate
Volumetric flask (250 cm3) 1
Beaker (250 cm3) 1
Weighing bottle 1
Funnel (small) 1
Wash bottle for distilled water 1
Burette stand 1
Solutions Provided
Oxalic acid solution: About M/100, prepared by dissolving 0.35 g of oxalic acid
dihydrate in water and making up the volume to 250 cm3 in a volumetric flask.
Approximately M/250 solution of potassium permanganate, prepared by

dissolving 0.20 g of potassium permanganate in distilled water and making up the
solution to 250 cm3. It is then stored in a dark place preferably in an amber coloured 51
bottle for a few days. Potassium permanganate solution is stored in dark because
light accelerates decomposition of KMnO4 by the reaction given below:
4 KMnO 4 + 2 H2O 4 MnO2 ↓ + 4 KOH+ 3 O2 ↑
Indicator
KMnO4 acts as a self indicator, so no other indicator is required.
2.4.3 Procedure
As indicated above, you are provided with approximately M/250 potassium
permanganate solution, and a solution of oxalic acid which is to be estimated. You
are required to prepare a standard solution of ferrous ammonium sulphate,
which is also known as Mohr’s salt.
There are three steps in the experiment as follows:
1. Preparation of a standard solution of ferrous ammonium sulphate
2. Standardisation of given KMnO4 solution by titrating against the standard ferrous

ammonium sulphate solution
3. Titration of given oxalic acid solution with the standardised KMnO4 solution
1. Preparation of standard ferrous ammonium sulphate solution

(concentration ═ M/50)
Take approximate mass of a glass weighing bottle. Then weigh it accurately with
about 1.956 g of Mohr's salt. Transfer the salt to a clean and dry volumetric flask
of 250 cm3 capacity through a glass funnel. Find out the accurate mass of the
bottle after transferring Mohr's salt. The difference between the two masses gives
the actual amount of Mohr's salt transferred. Record these values in your
observation note book. To the contents of the volumetric flask add about
10 cm3 of dilute H2SO4 (1 M) and about 50 cm3 of distilled water, dissolve the
salt completely; add more water, if necessary. Finally, make the volume upto the
mark by adding distilled water carefully.
Caution: If the solution turns brownish, then the amount of added acid
is not sufficient. Discard this solution. Do the whole exercise again
using more H2SO4.
2. Standardisation of potassium permanganate solution
Fill up the burette with the given KMnO4 solution and mount the burette on a
stand; also insert a parallex card. Note the reading in the burette and record it in
the Observation Table I. Pipette out the standard Mohr’s salt solution (20 cm3)
In case of potassium in a conical flask and add an equal volume of dil. sulphuric acid (5M). For this
permanganate it is purpose take a test tube and fill it a little more than half with H2SO4 and
convenient to use the mark its level so that you add the same amount of H2SO4 in every
upper meniscus. titration. Titrate against KMnO4 till a light but permanent pink colour is
obtained. This indicates the end point of the titration. Note the burette reading
and record it in Observation Table I. The difference of two readings gives a
rough estimate of the volume of KMnO4 required. Repeat the titration to get at
52 least two concordant readings to ensure a correct and exact measurement.
3. Titration of given oxalic acid solution against standardised KMnO4
solution
Pipette out the given oxalic acid solution (20 cm3), add equal volume of dil.
H2SO4, heat to 60°C-70° C and titrate against KMnO4 solution as above.
Repeat the titration till two concordant readings are obtained. Record the
readings in Observation Table II.
SAQ 4
Can HCl or HNO3 be used in place of H2SO4 for making the medium acidic in a
redox titration where KMnO4 is used as an oxidant? Justify your answer.
2.4.4 Observations
Approximate mass of the weighing bottle ═ m1 ═ ..g
Mass of bottle + Mohr’s salt ═ m2 ═ ..g
Mass of bottle (after transferring the Mohr’s salt) ═ m3 ═ .g
Mass of Mohr’s salt transferred ═ m2 ─ m3 ═ m ═ ...g
Molar mass of Mohr’s salt ═ 392.15 g mol─1
Volume of Mohr’s salt solution prepared (V) ═ 250 cm3
Molarity of Mohr’s salt solution (M1)
m × 1000
mol dm −3
Molar mass × 250
m×4
mol dm −3 = .......... .mol dm− 3
392.15
Observation Table I: Titration of standard Mohr’s salt solution vs.

Potassium permanganate solution
Sl. Volume of Mohr’s salt Burette reading Volume of KMnO4
3 3
No. solution (cm ) (cm )
Initial Final (Final ─ Initial)
1 20
2 20
3 20
Observation Table II: Titration of given Oxalic acid solution vs. Potassium
permanganate solution
Sl. Volume of oxalic acid Burette reading Volume of KMnO4
3 3
No. (in cm ) (in cm )
1 20
2 20
3 20 53
2.4.5 Calculations
Estimation of the strength of potassium permanganate
Molarity of Mohr’s salt solution ═ M1 ═ .. mol dm−3
Vol. of Mohr’s salt solution used ═ V1 ═ ..20 cm3
Vol. of KMnO4 solution used (from Table I) ═ V2 ═ cm3
Molarity of KMnO4 solution ═ M2 ═ ?
Using the molarity equation,
M1V1 ═ 5 M2 V2
M1V1
Molarity of KMnO4 solution, M 2 = = .......... .moldm − 3
5V2
Estimation of strength of given oxalic acid solution
Molarity of KMnO4 ═ M3 ═ M2 ═ mol dm−3 or M
Vol. of KMnO4 solution used ═ V3 ═ ...........cm3
Vol. of oxalic acid solution taken ═ V4 ═ 20 cm3
Molarity of oxalic acid ═ M4 ═ ?
2M4V4 ═ 5 M3V3
5M 3V3
Molarity of oxalic acid solution, M 4 = = mol dm − 3
5V4
Strength of oxalic acid solution M 4 × 126 g dm −3
= .......... g dm −3
2.4.6 Result
The strength of given oxalic acid solution ═ .. g dm−3
The above value can be compared with the actual one which you get from
your counsellor.
2.5 ANSWERS
1) i) H2 → 2 H+ + 2e −
ii) S−2 → S + 2e−
iii) Cl 2 + 2e → 2Cl −
54 iv) Cu + → Cu 2 + e −
v) −2
O 2 + 2e → 2O
2) 2KMnO 4 + 3H2SO 4 → K 2SO 4 + 2 MnSO 4 + 3H2O + 5[O]
[2FeSO 4 + H2SO 4 + (O) → Fe 2 (SO 4 )3 + H2O] × 5
2KMnO 4 + 10 FeSO 4 + 8H2SO 4 → K 2SO 4 + 2MnSO 4 + 5Fe 2 (SO 4 )3 + 8H2O
3) KMnO4 is not pure and always contains some of its reduction products
such as MnO2; also it is easily decomposed by reducing agents present in
H2O.
4) HCl acts a reducing agent, itself getting oxidised to Cl2; and HNO3 acts as
an oxidizing agent again competing with the action of KMnO4. Therefore,
neither of these can replace H2SO4.
55
EXPERIMENT 3
ESTIMATION OF WATER OF
CRYSTALLISATION IN
MOHR’S SALT
Structure
3.1 Introduction 3.4 Procedure
Expected Learning Outcomes 3.5 Observations
3.2 Principle 3.6 Calculations
3.3 Requirements 3.7 Result
3.1 INTRODUCTION
In the previous experiment you have learnt about and performed a redox
titration based on permanganatometry. In that titration you determined the
strength of a reducing agent- oxalic acid by titrating against KMnO4 in the
acidic medium. The KMnO4 solution was standardised by titrating against a
standard solution of Mohr’s salt-another reducing agent. In the third
experiment you are going to perform another titration based on
permanganatometry to determine the number of water of crystallisation in
Mohr’s salt. In the next experiment you would learn about and perform another
type of redox titration namely, chromatometry in which you would be using
K2Cr2O7 as the oxidising agent.

state and explain the principle involved in permanganatometry;
prepare a standard solution of oxalic acid;
standardise the given solution of potassium permanganate; and
determine the number of water of crystallisation in Mohr’s salt by

permanganatometry.
56
Experiment 3 Estimation of Water of Crystallisation in Mohr’s Salt
3.2 PRINCIPLE
In the determination of the water of crystallisation in Mohr’s salt (hydrated
ferrous ammonium sulphate) we first prepare its solution of known
concentration (g dm−3). Then we estimate it as anhydrous ferrous ammonium
sulphate by permanganatometry. The strength (g dm−3) so obtained and
actually taken is then used to determine the water of crystallisation as follows.
Let there be ‘x’ number of water of hydration in Mohr’s salt, the molar mass of
Mohr’s salt can be written as follows:
Molar mass of Mohr’s salt = Molar mass of the anhydrous salt + 18x
= 284 +18x
Let the amount of Mohr’s salt taken to prepare the solution = A g
Let the amount of anhydrous salt determined by permanganatometry = B g
Then we can write the equation as follows:

A 284 + 18x
= (3.1)
B 284
By knowing A and B, the equation can be solved to get the value of x, the
number of water of hydration in Mohr’s salt. Let us now take up the
determination of water of crystallisation in Mohr’s salt by permanganatometry.
As discussed in the previous experiment, potassium permanganate is an oxidising

agent and gets reduced in the presence of a suitable reducing agent. In this
experiment we would be titrating it with Mohr’s salt –a reducing agent. Permanganate
ions, MnO −4 get reduced in acidic, neutral or alkaline medium. In acidic medium, it gets
reduced to Mn2+ ion as per the following equation.
M nO 4− + 8H+ + 5e − M n2 + + 4H2O (3.2)
On the other hand, the Fe (II) ions provided by Mohr’s salt (ferrous ammonium
sulphate) get oxidised to Fe (III) as per the following equation
Fe2 + → Fe3 + + e − (3.3)
The overall ionic equation for the titration of Mohr’s salt and permanganate ion in
acidic medium can be obtained by adding Eq. 3.2 and Eq. 3.3 after balancing the
number of electrons between them as follows:
[Fe 2 + → Fe 3 + + e − ] × 5
MnO −4 + 8H + + 5 Fe + → Mn 2 + + 4H2O
MnO −4 + 8H + + 5 Fe + → Mn 2 + + 4H2O
(3.4)
As per equation 3.4, one mole of permanganate ions oxidises 5 moles of
ferrous ions in the acidic medium. The corresponding molarity equation would
be as given below:
MKMnO4VKMnO4 1
= (3.5)
MFe(II)VFe(II) 5
57
MKMnO4VKMnO4 = MFe(II) VFe(II) (3.6)
As you know, KMnO4 is a secondary standard and has to be standardised

against a primary standard. We would use oxalic acid as the primary standard
in this experiment. The oxalate ions get oxidized to CO2 as per the following
equation:
C2O24− → 2CO2 + 2e − (3.7)
The overall reaction between permanganate ion and oxalate ion can be
obtained by combining Eq. 3.2 and Eq. 3.7 with suitable balancing of electrons
as follows:
[Mn O4− + 8 H+ + 5e − Mn2 + + 4 H2O] × 2

[C2O24− 2CO2 + 2e− ] × 5 (3.8)
2 Mn O−4 + 16H+ + 5C2O24− 2 Mn2 + 10 CO2 + 8 H2O
As per equation 3.8, two moles of permanganate ions oxidise 5 moles of

oxalate ions in the acidic medium. The corresponding molarity equation would
be as given below:
MPermangana te VPermangana te 2
= ...(3.9)
M oxalate Voxalate 5
5MPermangana te VPermangana te = 2Moxalate Voxalate (3.10)
3.3 REQUIREMENTS
Apparatus Chemicals
Burette (50 cm3) 1 Ferrous ammonium sulphate
Pipette (20 cm3) 1 Oxalic acid

Potassium permanganate
Conical flasks (250 cm3) 2
Sulphuric acid (1 M)
Test tube 1
Beaker (250 cm3) 1
Weighing bottle 1
Funnel (small) 1
Wash bottle 1
Burette stand and clamp 1
Solutions Provided
Mohr’s salt solution (8 g dm−3): Prepared by taking accurately weighed about 8 g

of Mohr’s salt and transferring it to a volumetric flask of 1 dm3 capacity. To this
58 about 10 cm3 of dilute H2SO4 (1 M) and about 50 cm3 of distilled water is added to
dissolve the salt completely. The volume is then made up by adding more distilled
water to the volumetric flask.
Approximately M/250 solution of potassium permanganate: Prepared by

dissolving 0.16 g of potassium permanganate in distilled water and making up the
volume of the solution to 250 cm3. It is then stored in a dark place preferably in an
amber coloured bottle for a few days. Potassium permanganate solution is stored in
dark because light accelerates decomposition of KMnO4 by the reaction given
below:
4 KMnO 4 + 2 H2O → 4 KMnO 2 ↓ +4 KOH + 3 O 2
Indicator
KMnO4 acts as a self indicator, so no other indicator is required.
3.4 PROCEDURE
As indicated above, you are provided with approximately M/250 potassium
permanganate solution, and a solution of Mohr’s salt which is to be estimated. You
are required to prepare a standard solution of oxalic acid
There are three steps in the experiment.
1. Preparation of a standard solution of oxalic acid
2. Standardisation of given KMnO4 solution by titrating against the standard

solution of oxalic acid
3. Titration of given Mohr’s salt solution with the standardised KMnO4 solution
1. Preparation of standard oxalic acid solution (M/20)
Take approximate mass of a glass weighing bottle. Then weigh it accurately

with about 0.32 g of oxalic acid. Transfer it to a clean and dry volumetric flask of
250 cm3 capacity through a glass funnel. Find out the accurate mass of the
bottle after transferring oxalic acid. The difference between the two masses
gives the actual amount of oxalic acid transferred. Record these values in your
observation note book. To the contents of the volumetric flask add about
30-40 cm3 of distilled water, dissolve the acid completely; add more water, if
In case of potassium
necessary. Finally, make the volume upto the mark by adding distilled water permanganate it is
carefully. convenient to use the
upper meniscus.
2. Standardisation of potassium permanganate solution
Fill up the burette with the given KMnO4 solution and mount the burette on a
stand; also insert a parallex card. Note the reading in the burette and record it
in the Observation Table I. Pipette out the standard oxalic acid solution
(20 cm3) in a conical flask and add an equal volume of dil. sulphuric acid (1M)
heat to 60°-70° C. For this purpose take a test tube and fill it a little more
than half with H2SO4 and mark its level so that you add the same
amount of H2SO4 in every titration. Titrate against KMnO4 till a light but
permanent pink colour is obtained. This indicates the end point of the titration.
59
Note the burette reading and record it in Observation Table I. The difference of
two readings gives a rough estimate of the volume of KMnO4 required. Repeat
the titration to get at least two concordant readings to ensure a correct and
exact measurement.
3. Titration of given Mohr’s salt solution against standardised KMnO4

solution
Pipette out the given Mohr’s salt solution (20 cm3), add equal volume of dil.
H2SO4 and titrate against KMnO4 solution as above. Repeat the titration till
three concordant readings are obtained. Record the readings in Observation
Table II.
3.5 OBSERVATIONS
Mass of bottle + oxalic acid (before transferring) ═ m2 ═ ..g
Mass of bottle (after transferring the oxalic acid) ═ m3 ═ .g
Mass of oxalic acid transferred ═ m2 ─ m3 ═ m ═ ...g
Molar mass of oxalic acid ═ 126.06 g mol─1
Volume of oxalic acid solution prepared (V) ═ 250 cm3
Molarity of oxalic acid solution (M1)

m × 1000 m×4
= mol dm − 3 = mol dm− 3
Molar mass × 250 126.06
Observation Table I: Titration of standard oxalic acid solution vs.

Sl. Volume of Oxalic acid Burette reading Volume of KMnO4

3 3
No. solution (in cm ) in (cm )
1 20
2 20
3 20
Observation Table II: Titration of given Mohr’s salt solution vs.

Sl. Volume of Mohr’s salt Burette reading Volume of KMnO4

3 3
solution (in cm ) (in cm )
No.
Initial Final
(Final ─ Initial)
1 20
2 20
3 20
60
3.6 CALCULATIONS
Estimation of the strength of potassium permanganate
Molarity of oxalic acid solution ═ M1 ═ .. mol dm −3
Vol. of oxalic acid solution used ═ V1 ═ ..20 cm3
Vol. of KMnO4 solution used (from Table I) ═ V2 ═ cm3
Molarity of KMnO4 solution ═ M2 ═?
Using the molarity equation (Eq. 3.10),

5MPermangana te VPermangana te = 2Moxalate Voxalate
5M2V2 = 2 M1V1
Molarity of KMnO4 solution
2 M1V1
M2 = = ..mol dm−3
5V2
Estimation of strength of given Mohr’s salt solution

Molarity of KMnO4 ═ M3 ═ M2 ═ mol dm−3 or M
Vol. of KMnO4 solution used ═ V3 ═ cm3
Vol. of Mohr’s salt solution taken ═ V4 ═ 20 cm3
Molarity of Mohr’s salt ═ M4 ═?
Using the molarity equation (3.6),
5MPermangana te VPermangana te = MMohr's salt VMohr's salt
5M3 V3 ═ M4 V4
Molarity of Mohr’s salt solution
5M3V3
M4 = = .......... ... mol dm−3
V4
Strength of Mohr’s salt (anhydrous) = M4 × 284 g dm−3
= B = _______ g dm−3
Determination of number of water of crystallisation

The amount of Mohr’s salt taken to prepare the solution = A g
(Your counsellor would tell you the amount of Mohr’s salt actually taken)
The amount of anhydrous salt determined by permanganatometry = B g
According to Eq.3.1
A 284 + 18 x
= (3.1)
B 284
Substitute the values of A and B and solve for x. 61
3.7 RESULT
The number of water of hydration in Mohr’s salt = .
The above value can be compared with the actual one given on the bottle of
Mohr’s salt.
62
Experiment 4 Estimation of Iron (II) Ions by Chromatometry using Internal Indicator
EXPERIMENT 4
ESTIMATION OF IRON(II)
IONS BY CHROMATOMETRY
USING INTERNAL INDICATOR
Structure
4.1 Introduction 4.5 Observations
Expected Learning Outcomes 4.6 Calculations
4.2 Principle 4.7 Result
4.3 Requirements 4.8 Answers
4.4 Procedure
4.1 INTRODUCTION
You have studied about the concept of redox titrations and performed one such type
of titration called permanganatometry in Experiment 2. In permanganatometric titration
you used potassium permanganate as the oxidising agent to estimate the strength of
given oxalic acid solution. In the fourth experiment you will perform another redox titration
which makes use of potassium dichromate as the oxidising agent. This type of titration is
called chromatometry. Dichromate acts as an oxidising agent only in the acidic
medium. The general theory behind chromatometry is the same as that for
permanganatometry. The only difference being that while KMnO4 acts as a self
indicator, an indicator has to be used in chromatometry. The relevant equations
along with the potentials are given in the following sections. The theory behind the
colour change of indicators is also explained.

define chromatometry as a redox titration;
state and explain the principle of chromatometry;
list the apparatus and chemicals required for the estimation of Fe(II) ions in the
given solution by chromatometry;
63
record the observations of the titration performed;
calculate the molarity and the strength of the given Fe(II) solution; and
report the result of the titration performed.
4.2 PRINCIPLE
Potassium dichromate is an oxidising agent in acidic medium and reacts according to
the following half-reaction, to give chromium (III) ions as the reduction product.
Cr2O72 − + 14H + + 6e − + 2Cr 3 + + 7H2O, (E 0 = 1.33 V)

(4.1)
( + 6 state) ( + 3 state)
Fe2+ ions when titrated with dichromate get oxidised to Fe3+ as per the following
equation.
(cf. Eq. 2.1 in Experiment 2):
Fe 2 + Fe3 + + e − (E 0 = 0.77 V) (2.1)
The overall ionic equation of the titration can be obtained by adding

Eq. 4.1 and Eq. 2.1 after balancing the number of electrons between them
as follows:
[Fe 2 + Fe 3 + + e − ] × 6 (4.2)
Cr2O72 − +
+ 14H + 6e 2Cr 3+
+ 7H2O (4.1)
Cr2O72 − + 6Fe 2 + + 14H 2Cr 3 + + 7H2O (4.3)
We see from Eq. 4.3 that one mole of potassium dichromate reacts with 6 moles of
iron (II) ions in solution. Therefore, substituting the values of p and q in the molarity
equation, the molarities are related as:
MDVD 1
=
MFeVFe 6
or 6MDVD = MFeVFe (4.4)
where MD and MFe represent the molarities of potassium dichromate and iron (II)
solutions and VD and VFe represent their volumes, respectively.
As you know, the factor 6 signifies that each mole of potassium dichromate reacts
quantitatively with 6 moles of iron (II).
1 mole K 2Cr2O7 = 6 moles Fe (II)
Potassium dichromate does not oxidise HCl, whereas KMnO4 oxidises it to Cl2.
Hence, KMnO4 cannot be used as an oxidising agent in estimating metal ions
where the solution is made using HCl, whereas K2Cr2O7 can be used in such
cases too. Since, metals are leached out from their minerals with HCl in many
64 cases, chromatometry is the preferred technique of estimation in such cases.
Indicator
A dilute solution of K2Cr2O7 has a faint orange colour and chromium (III) obtained
as the reduction product is green in colour. So, a drop of K2Cr2O7 in excess at the
end point, unlike KMnO4, is not sufficient to give a distinct colour to the solution.
Further, the green colour of chromium (III) ions produced also interferes, therefore,
K2Cr2O7 cannot be used as a self indicator. A redox indicator must, therefore, be
used.
Diphenylamine was one of the first internal redox indicators used in this titration.
However, as it is sparingly soluble in water, sodium salt of diphenylamine sulphonic
acid can be used instead, as it is water soluble. These indicators are colourless in
the reduced form and become intensely coloured on oxidation (deep blue-violet
with diphenylamine; red-violet with sodium diphenylamine sulphonate).
At potentials lower than 0.73 V, iron is in +2 state and the indicator is present in
form I which is colourless. In the course of the titration, when Fe2+ gets converted to
Fe3+, the potential increases gradually. At equivalence point all the ferrous ions are
oxidised and as discussed earlier, at this stage there is a jump in the potential. This
increase in the redox potential of the solution is sufficient to cause the oxidation of
the indicator to form II. Form II gets readily converted into form III, which has a
distinct colour and marks the end point. The above description about the indicator
can be represented as shown below:
Oxidation + −
2 N N N + 2H + 2e
H H H
Diphenylamine(I) Diphenylbenzidine(II)
(colourless) (colourless)
+ + −
N N + 2e
H H
Diphenylbenzidine(III) (violet)
SAQ 1
a) Write the chemical reaction involving a titration of K2Cr2O7 and FeSO4 solution
in presence of dilute H2SO4. [Hint: The reaction involves two steps].
b) Explain why K2Cr2O7 reacts as an oxidising agent only in the acidic medium.
SAQ 2
In the following put a tick mark (√ ) on the correct and cross (×) on the wrong
statements.
K2Cr2O7 is superior to KMnO4 because:
i) K2Cr2O7 is more stable than KMnO4 both in the dry state as well as in solution.
ii) A solution of K2Cr2O7 is not intensely coloured.
iii) K2Cr2O7 is not reduced by cold HCl, if acid concentration does not
exceed 1M or 2M.
iv) K2Cr2O7 can be easily weighed.

65
4.3 REQUIREMENTS
Apparatus Chemicals
Burette (50 cm3) 1 Ferrous ammonium sulphate
Pipette (20 cm3) 1 Sulphuric acid, dilute (1 M)
Conical flask (250 cm3) 1 Phosphoric acid (85%)
Beaker (250 cm3) 1
Weighing bottle 1
Funnel (small) 1
Volumetric flask (250 1

cm3)
Wash bottle 1
Burette stand 1
Solutions Provided
Ferrous ammonium sulphate solution (unknown concentration): Prepared

by dissolving about 8.0 g of ferrous ammonium sulphate in distilled water and
dilute H2SO4 and making up the volume to 1 dm3.
Approximately M/300 solution of potassium dichromate: Prepared by

dissolving 0.245 g of potassium dichromate in distilled water and making up the
volume to 250 cm3.
Diphenylamine (1%) solution in concentrated sulphuric acid or sodium

diphenylamine sulphonate (0.2% aqueous solution).
4.4 PROCEDURE
Potassium dichromate, as mentioned earlier, can be used as a primary
standard which means that a standard solution of dichromate can be made by
weighing an exact amount of the substance, dissolving it in water and making
up to the known volume with distilled water. However, in this particular
experiment, we are provided with solution of about M/300 solution of K2Cr2O7,
which would act as an intermediate and needs to be standardised with a
standard solution of ferrous ammonium sulphate.
This experiment has three steps as given below:
1. Preparation of standard solution of ferrous ammonium sulphate
2. Standardisation of given solution of potassium dichromate with standard

solution of ferrous ammonium sulphate
3. Titration of given solution of ferrous ammonium sulphate with the

standardised solution of potassium dichromate
66
1. Preparation of M/50 ferrous ammonium sulphate solution
Take approximate mass of a glass weighing bottle. Then weigh it

accurately with about 1.956 g of Mohr's salt. Transfer the salt to a clean
and dry volumetric flask of 250 cm3 capacity through a glass funnel. Find
out the accurate mass of the bottle after transferring Mohr's salt. The
difference between the two masses gives the actual amount of Mohr's salt
transferred. Record these values in your observation note book. To the
contents of the volumetric flask add about 10 cm3 of dilute H2SO4 (1M) and
about 50 cm3 of distilled water, dissolve the salt completely; add more
water, if necessary. Finally, make the volume upto the mark by adding
distilled water carefully.
Caution: If the solution turns brownish, then the amount of acid

added is not sufficient. Discard this solution. Do the whole exercise
again using more H2SO4.
2. Standardisation of potassium dichromate solution
Pipette out 20 cm3 aliquot of standard ferrous ammonium sulphate solution

into a 250 cm3 conical flask. Add approximately 20 cm3 of dilute sulphuric
acid (1 M), 5 cm3 of phosphoric acid and 5 to 10 drops of the indicator
solution. Titrate this with the dichromate solution. As the titration proceeds,
the colour changes to pale green, then to a greyish blue-green and with
one drop to a persistent deep blue-violet colour of the indicator in the
oxidised form. Record the volume of the titrant accurately.
Repeat the titration to get at least two concordant readings to ensure a

correct and exact measurement. Record the observations in Observation
Table I.
3. Titration of the given ferrous ammonium sulphate solution against

standardised K2Cr2O7 solution
Perform this titration exactly in the same manner as in the above

experiment by taking the given ferrous ammonium sulphate solution instead
of standard ferrous ammonium sulphate solution. Record the observations
in Observation Table II.
4.5 OBSERVATIONS
Mass of bottle + ferrous ammonium sulphate ═ m2 ═ ..g

(before transferring salt)
Mass of bottle (after transferring the salt) ═ m3 ═ .g
Mass of ferrous ammonium sulphate transferred ═ m2 ─ m3 ═ m ═ g
Molar mass of ferrous ammonium sulphate ═ 392.15 g mol─1
Volume of ferrous ammonium sulphate solution prepared (V) ═ 250 cm3
Molarity of ferrous ammonium sulphate solution (M1) 67

M × 1000
mol dm − 3
Molar mass × 250
M×4
= mol dm − 3 = ...... mol dm − 3
392.15
Observation Table I: Titration of standard FAS solution vs. Potassium

dichromate solution
Sl. No. Volume of FAS Burette reading Volume of K2Cr2O7

3
solution (cm ) 3
(cm )
Initial Final (Final — Initial)
1 20
2 20
3 20
Observation Table II: Titration of given FAS solution vs. Potassium

dichromate solution
Sl. No. Volume of FAS Burette reading Volume of K2Cr2O7

3
solution (cm ) 3
(cm )
Initial Final (Final — Initial)
1 20
2 20
3 20
4.6 CALCULATIONS
Standardisation of K2Cr2O7
Molarity of FAS solution = M1 = mol dm−3
Volume of FAS solution taken = V1 = 20 cm3
Volume of K2Cr2O7 solution used (from Table I) = V2 = cm3
Molarity of K2Cr2O7 solution = M2 = ?
M1V1 = 6 M2V2
Molarity of K2Cr2O7
M1V1 −3
M2 = = .......... .. mol dm
6V2
Estimation of strength of given Iron (II) solution (FAS)
Molarity of K2Cr2O7 solution = M3 = M2 = mol dm−3
Volume of K2Cr2O7 solution used = V3 = 20 cm3
Volume of iron(II) solution taken = V4 = 20 cm3

68
Molarity of iron(II) solution = M4 =?
6M 3V3
M4 =
V4
Molarity of Fe(II) ions = .. mol dm−3
Strength of iron(II) ions in the given FAS solution
= Molarity of iron (II) ions in the given solution × molar mass
= M4 × 392.15 g dm−3
= gdm−3
4.7 RESULT
The strength of Fe(II) ions in the given FAS solution is found to be = g dm−3
The above value can be compared with the actual one which you can get from your
counsellor.
4.8 ANSWERS
1) a) K 2Cr2O7 + 4 H2SO 4 → K 2SO 4 + Cr2 (SO 4 )3 + 4 H2O + 3[O]
[2 FeSO 4 + H2SO 4 + (O) → Fe2 (SO 4 )3 + H2O] × 3
K 2Cr2O7 + 6 FeSO 4 + 7 H2SO 4 → 3 Fe2 (SO 4 )3 + K 2SO 4 + 7 H2O
b) In presence of an alkali, potassium dichromate reacts to give

chromate salt and cannot act as an oxidising agent.
2) i) √
ii) ×
iii) √
iv) ×
69
EXPERIMENT 5
ESTIMATION OF COPPER(II)
IONS IODOMETRICALLY
Structure
5.1 Introduction Principle
5.2 Iodimetry and Iodometry Procedure
Indicator in Titrations Involving Iodine Observations
Standardisation of Sodium Thiosulphate Calculations
5.3 Estimation of Cu (II) Ions Result

Iodometrically using Sodium
5.4 Answers
Thiosulphate
5.1 INTRODUCTION
In the previous three experiments, you have performed two types of redox
titrations, namely, permanganatometry and chromatometry. In this experiment,
you would estimate the amount of copper (II) ions in a given sample solution
using iodine as the reducing agent. You will realise that this experiment is
also based on the principle of redox reactions. The redox reactions that
make use of iodine can take place in two ways as described in the next
section. These titrations make use of I2/I− redox reaction and the end point is
detected by using starch as an indicator. The theory behind the iodometric
estimation of cupric ions, Cu2+ is given along with the procedural details of the
experiment.

define and differentiate between iodometry and iodimetry;
state and explain the principle of iodometry;
record the observations of the experiment;

70
Experiment 5 Estimation of Copper(II) Ions Iodometrically
2+
use the iodometric method in estimating Cu ions; and
calculate the strength of the Cu(II) ions in the given solution.
5.2 IODIMETRY AND IODOMETRY

Iodine is a mild oxidising agent and in the presence of a suitable reducing
agent, it gets reduced to iodide ions, I− according to the following equation:
I2 + 2e− 2I−, E0 = 0.54 V ... (5.1)
On the other hand, a number of oxidising agents can oxidise I− ions into I2. In
fact, both these reactions are made use of in analytical chemistry. Titrations
involving the use of I2 as a titrant to estimate the reducing agents are termed
as iodimetric titrations. However, iodine, being a much weaker oxidising agent
lodimetric titrations are
than potassium permanganate and potassium dichromate, has limited
used for estimating
applicability. Moreover, it is very volatile in nature and also has poor solubility.
reducing agents while
In certain cases, the oxidising agent to be determined is mixed with an excess iodometric titrations are
used for oxidising
of potassium iodide, KI, and kept for some time. The iodine, liberated during
agents.
the reaction, is titrated against a standard solution of a reducing agent, e.g.,
sodium thiosulphate, Na2S2O3. These titrations are referred to as iodometric
titrations. Since, Cu2+ ions, can behave as an oxidising agent by getting
reduced to Cu+ ions, we can use iodometric method for their determination.
lodimetry: Titration with
Ideally an iodometric titration should be a titration using KI as a titrant to titrate iodine
the oxidising agent. In such a reaction, more and more of iodine is liberated
from iodide ions as the titration proceeds. The end point of such a titration
lodometry: Titration of
would be a stage where the liberation of iodine ceases. It is impossible to
iodine produced by a
detect this end point with the help of an indicator. Starch can be used to detect chemical reaction
the 'just appearance' or the 'just disappearance' of iodine but not the cessation
of I2 formation.
An indirect method of end point determination becomes essential in such

cases. A known amount of the solution of the oxidising agent (to be
determined) is measured and mixed with an excess of a solution of KI and
acid. The solution is then left for about five minutes in the dark for the reaction
to complete and the liberated iodine is titrated with a standardised solution of
sodium thiosulphate using starch as the indicator. The following reaction
takes place:
I2 + Na2S2O3 2NaI + Na2S4O6 (5.2)
An excess of KI is used because iodine has got very poor solubility in water.
Iodine forms an unstable complex, KI3 with KI which is readily soluble in H2O.
KI2 + I2 K+[I3]−
In fact, iodine in an aqueous solution containing KI exists mainly as the triiodide

ion, I3− and there is an equilibrium between I3− ion and I2. In the course of the
titration, as I2 is consumed, more and more of I3− ions dissociate to give I2,
which reacts with the thiosulphate. Further, such a titration should be carried
out in cold, as I2 is volatile and also the indicator, starch, loses its sensitivity at
high temperatures. 71
SAQ 1
Give two limitations of I2 as a titrant.
5.2.1 Indicator in Titrations Involving Iodine

In principle, iodine can be used as a self indicator like KMnO4, as a drop of
iodine can impart a pale yellow colour to a solution. As the colour imparted by
iodine is quite faint, in practice, it becomes difficult to use this as an indication
of the end point. Iodine is known to form a blue coloured adsorption complex
with starch. This property of starch is exploited in using it as an indicator for
titrations involving iodine.
The use of starch

In an iodometric determination, we titrate I2 with S2O32 − ions and at the end
enhances the sensitivity point, addition of one drop of S2O32 − ions should just decolourise the blue
of the determination of colour of starch-iodine complex. In such titrations, starch should be added just
the end point.
before the end point, when a very little amount of I2 remains and the solution
being titrated has a faint straw yellow colour. If starch is added earlier, i.e.,
when a large amount of iodine is present, a large amount of starch-iodine
complex is formed. This complex reacts quite slowly with S2O32 − and it is likely
that the solution is over titrated.
5.2.2 Standardisation of Sodium Thiosulphate

As explained above, in iodometry, we titrate the liberated iodine with a
standardised solution of sodium thiosulphate. Though sodium thiosulphate,
Na2S2O3.5H2O, can be obtained chemically pure, a standard solution of
thiosulphate cannot be made by exact weighing. This is because thiosulphate
reacts with atmospheric O2 and also with the CO2 dissolved in water. More so,
even some microorganisms can decompose thiosulphate. Therefore, we do
not use thiosulphate as a primary standard and its solution is standardised.
A number of oxidising agents are available for the standardisation of Na2S2O3.

Potassium dichromate is normally used for the purpose.
In acidic medium Cr 2O72− ion gets reduced to Cr (III) as shown in the following
equation:
Cr 2O72 − + 14 H+ + 6e − 2 Cr 3 + + 7 H2O (5.3)
and iodide ions from KI get oxidised to I2:
2I− I2 + 2e−
To maintain electron balance, multiplying the above equation by 3, we get,
6I− 3I2 + 6e− (5.4)
The overall ionic equation for the titration can be obtained by adding Eq.5.3
and Eq. 5.4,
72 Cr2O72 − + 14 H+ + 6 I− → Cr 3 + + 3 I2 + 7 H2O (5.5)

We see from Eq. 5.5 that one mole of potassium dichromate reacts with
6 moles of potassium iodide liberating 3 moles of iodine.
The liberated iodine, in turn, reacts with sodium thiosulphate solution

as,
2S2O32 − S 4O 26− + 2e − (5.6)
I2 + 2e − 2I− (5.1)
Since three moles of I2 are liberated by one mole of Cr2O72−
3I2 + 6e − 6I− (5.7)
The overall ionic equation for the titration of liberated I2 with sodium thiosulphate
can be obtained by adding Eq. 5.6 (multiplied by three) and Eq. 5.7,
[2S2O32 − S 4O26 − + 2e − ] × 3 (5.6)

− −
3I2 + 6e 6I (5.7)
3I2 + 6S2O32 − → 6I− + 3S 4O26 − (5.8)
The net chemical reaction involving a titration of potassium dichromate and

sodium thiosulphate in the presence of excess potassium iodide can be
written by combining Eq. 5.5 and Eq. 5.8,
Cr2O32 − + 14 H+ + 6 I− → 2 Cr 3 + + 3 I2 + 7 H2O (5.5)

3I2 + 6S2O32 − → 6I + −
3S 4O26 − (5.8)
Cr2O72 − + 14H+ + 6S 2O32 − → 2Cr 3 + + 3S 4O26 − + 7H2O (5.9)
We see from Eq. 5.9 that one mole of potassium dichromate is equivalent
to 6 moles of sodium thiosulphate. Therefore, substituting the values in
the molarity equation, the molarities are related by the following
Factor '6' here signifies
relationship. that one mole of K2Cr2O7
liberates 3 moles of I2
M1 V1 1
= which is equivalent to
M2 V2 6 6 moles of sodium
thiosulphate.
or 6M1V1 = M2V2
where M1 and M2 represent the molarities of potassium dichromate and

sodium thiosulphate solutions and V1 and V2 are their volumes, respectively.
5.3 ESTIMATION OF Cu(II) IONS IODOMETRICALLY

USING SODIUM THIOSULPHATE
To determine the strength of Cu(II) ions in a given solution, it can be titrated
against a standardised solution of sodium thiosulphate in the presence of an
excess of KI using starch as an indicator. The type of titration involved is called
iodometric titration. Let us learn the principle and the equations involved in the
titration. 73
5.3.1 Principle
The reaction between Cu2+ and Na2S2O3 in acidic medium, in the presence of
excess of KI, involves oxidation of S2O32− to S2O26− , tetrathionate ion and
reduction of Cu2+ to Cu+ . The reaction between Cu2+ and KI is given as,
2I− I2 + 2e − (5.10)
Cu2 + + e − Cu + (5.11)
Balancing the reaction between Cu2+ and potassium iodide by combining

Eq. 5.10 and Eq. 5.11, we get,
2I− I2 + 2e − (5.10)
2+ − +
Cu +e Cu ] × 2 (5.11)
2 Cu2 + + 2 I− → 2 Cu+ + I2 (5.12)
We see that two moles of Cu2+ react with two moles of potassium iodide and
liberates iodine. The liberated iodine reacts with sodium thiosulphate, in the
following manner:
2S 2O32 S 4O26 − + 2e − (5.6)

−
I2 + 2e 2I −
(5.7)
I2 + 2S 2O32 − → 2I− + S 4O26 − (5.8)
The net chemical reaction involving a titration of copper (II) and sodium
thiosulphate in the presence of excess potassium iodide can be written by
combining Eq. 5.12 and Eq. 5.8.
2Cu2 + + 2I− → 2Cu + + I2 (5.12)

I2 + 2 S2O32 − → 2I −
+ S 4O26 − (5.8)
2Cu2 + + 2S 2O32 − → 2Cu+ + S 4O26 − (5.13)
We see from Eq. 5.13, that two moles of copper (II) are equivalent to two moles
of sodium thiosulphate. In other words one mole of copper (II) is equivalent to
one mole of sodium thiosulphate.
Therefore, substituting the values of p and q in the molarity equation, the

molarities are related by the following relationship:
M 3 V3 1
=
M 4 V4 1
or M3V3 = M4V4
where M3 and M4 represent the molarities of sodium thiosulphate and copper(II)

solutions, and V3 and V4, the volumes of sodium thiosulphate and copper(II)
solutions, respectively.
74
According to the above discussion, the iodometric determination of Cu2+ ions is
based on the following reaction:
2 Cu2 + + KI 2 CuI + 4 K + + I2 (5.14)

(excess)
Where cupric ions are reduced to cuprous ions and iodide ions are oxidised to
iodine. A look at the standard reduction potentials of Cu2+ / Cu+ and I2 / I− couples,
Cu2 + + e − Cu+ E 0 = 0.17 V (5.15)
I2 + 2e − 2I− E 0 = 0.17 V (5.16)
suggests that the reaction represented by Eq. 5.14 should proceed in the reverse
direction, i.e. iodine should oxidise Cu+ to Cu2+, but actually the reaction occurs as
given in Eq. 5.14. The Cul formed during the reaction has a very low solubility in
water, therefore, the concentration of the reduced form, Cu+, is greatly reduced and
the potential of Cu2+/Cu+ couple becomes greater than that of I2 / 2I−. This explains
the actual course of reaction.
SAQ 2
Write the chemical equations involving a titration of copper (II) sulphate with
thiosulphate in presence of excess KI. [Hint: It involves two steps]
5.3.2 Requirements
Apparatus No. Chemicals
Burette (50 cm3) 1 Potassium dichromate
Pipette (20 cm3) 1 Dilute sulphuric acid
Conical flasks (250 cm3) 1 Potassium iodide
Beaker (250 cm3) 1 Glacial acetic add
Funnel (small) 1 Potassium ammonium thiocyanate
Volumetric flask (250 cm3) 1 Distilled water
Measuring cylinder (10 cm3) 1 Copper sulphate pentahydrate
Test Tube 1 Sodium thiosulphate
Wash bottle 1 Starch
Weighing bottle 1 Dil. ammonia solution
Burette stand 1
Solutions Provided
Procedures for the preparation of these solutions are given for the sake of
information. These solutions would be prepared by the counsellor. 75
Preparation of solution of Cu2+ ions
Weigh accurately 1.25 g of copper sulphate petahydrate and transfer it to a
250 cm3 standard flask. Dissolve it in 20 cm3 of water add 1-2 cm3 of glacial acetic
acid and make up to 250 cm3 with water to prepare the M /50 standard solution.
The solution of copper sulphate undergoes hydrolysis. To prevent this, mineral

acids like sulphuric acid is added to the solution but the reaction between KI and
CuSO4 to liberate iodine does not occur in the presence of any mineral acid.
Hence the acid added to copper sulphate solution is neutralised by adding a slight
excess of ammonia. The excess ammonia is then neutralised with acetic acid.
Sodium thiosulphate solution (approx. M /50)

About 5g of sodium thiosulphate crystals are dissolved in 1dm3 of water that has
been recently boiled and cooled. An amount of 0.2 g of sodium bicarbonate is
added as a preservative and the solution stored in a clean bottle. Sodium
thiosulphate solutions are somewhat unstable. Apart from oxygen and dissolved
CO2 they are easily attacked by air-borne bacteria with the liberation of sulphur. In
case any turbidity is observed, the solution should be discarded.
Starch Solution: About 100 cm3 distilled water is heated to boiling in a beaker.
While this is being heated, 0.5 g to 1 g of soluble starch is stirred with about
10 cm3 of distilled water to give a paste. The paste is stirred into the boiling water
and boiled gently for a few minutes and cooled. The solution should be almost
clear. It is kept in a stoppered bottle. Starch solution should be freshly prepared
before use.
Potassium Iodide Solution: Prepared by dissolving 5.0 g KI in 100 cm3 of distilled

water.
5.3.3 Procedure
The iodometric determination of a solution of Cu(II) ion involves three steps as
follows:
1. Preparation of standard solution of potassium dichromate
2. Standardisation of sodium thiosulphate solution by titrating against standard

solution of potassium dichromate
3. Titration of the given Cu(II) ion solution against the standardised solution of
sodium thiosulphate
The three steps are explained below.
1) Preparation of M/300 standard solution of potassium dichromate

Take approximate mass of a glass weighing bottle. Then weigh it
accurately with about 0.245 g of potassium dichromate. Transfer the salt to
a clean and dry volumetric flask of 250 cm3 capacity through a glass
funnel. Find out the accurate mass of the bottle after transferring
potassium dichromate. The difference between the two masses gives the
actual amount of potassium dichromate transferred. Record these values
in your observation note book. Finally, make the volume upto the mark by
76 adding distilled water carefully.
2) Standardisation of sodium thiosulphate solution
Pipette 20 cm3 of potassium dichromate solution in a 250 cm3 conical flask,
add 10 cm3 of dilute sulphuric acid and 1 g sodium hydrogen carbonate with
gentle swirling to liberate carbon dioxide. Sodium hydrogen carbonate
maintains an atmosphere of CO2 in the solution which displaces the air and
prevents the oxidation of iodide from air. The reaction:
4I− + O2 + 4H+ 2l2 + 2H2O is catalysed by light, heat and air. Then add
0.5 g potassium iodide or 10 cm3 of 5% Kl solution, swirl, cover the flask with a
watch glass and allow the solution to stand for 5 minutes in a dark place. Titrate
against sodium thiosulphate solution from the burette until the solution becomes
a light pale yellow in colour. Then add 2 cm3 starch solution and continue to
titrate until the blue colour of starch-iodine complex disappears on addition of a
drop of the titrant. The final solution will be green coloured because of the
presence of chromium (III) ions. Record the burette readings before and after
the titration in Observation Table I. Repeat the same exercise to get at least
two concordant readings.
3) Titration of copper (II) solution against standardised sodium

thiosulphate solution
Pipette out 20 cm3 of the given solution of Cu(II) ions into a 250 cm3 conical
flask, and add 0.5 g solid potassium iodide or 10 cm3 of 5% KI solution, swirl it
to dissolve; then titrate with the standardised sodium thiosulphate which is
taken in a burette. When the brown colour of iodine, becomes pale yellow, add
2 cm3 of fresh starch solution. The colour of the solution at this stage is deep
blue. Swirl the flask for about 15 seconds and complete the titration adding
sodium thiosulphate solution dropwise. During the titration, as CuI is formed, it
absorbs I3− on the surface, as a result the reaction of I2 with Na2S2O3 titrant is
very slow. Therefore, very close to the end point, when the colour is very light
blue, add 1 g potassium thiocyanate, KSCN. Thiocyanate added at this stage
reacts with Cul and forms CuSCN displacing iodine from the surface, making
it available for the reaction.
CuI + SCN− CuSCN + l−
However, if thiocyanate is added earlier during the titration, it will be slowly

oxidised to sulphate by iodine. At the end point, the blue colour of the solution
disappears and the precipitate appears white, or slightly grey, when allowed to
settle. After standing for a couple of minutes at the end point, the precipitate
should become pure white. Record the burette readings in Observation Table II.
Repeat the same exercise to get at least two concordant readings.
SAQ 3
During iodometric titrations, starch is added only towards the end of the titration.
Why?
SAQ 4
Why is sodium hydrogen carbonate or sodium bicarbonate added in the
standardisation of sodium thiosulphate using potassium dichromate as titrand?
77
5.3.4 Observations
Mass of weighing bottle = m1= ..g
Mass of bottle + potassium dichromate = m2= ..g
Mass of the bottle (after transferring = m3= ..g

K2Cr2O7)
Mass of potassium dichromate = m2 − m3 = m = ..g

transferred
Molar mass of potassium dichromate = 294.19 g mol−1
Volume of K2Cr2O7 prepared (V) = 250 cm3
Molarity of K2Cr2O7 solution = M1
= m×4
mol dm−3
Molar mass
= m×4
mol dm − 3
294.19
= .. mol dm−3
Observation Table I: Potassium dichromate solution vs. sodium

thiosulphate solution
Sl. Volume of K2Cr2O7 Burette reading Volume of Na2S2O3

3 3
No. acid solution (in cm ) solution in (cm )
Initial Final
(Final ─ Initial)
1 20
2 20
3 20
Observation Table II: Titration of given solution of Cu (II) ions vs.

standardised sodium thiosulphate solution
Sl. Volume of Copper (II) Burette reading Volume of Na2S2O3

3 3
No. solution (in cm ) solution in (cm )
Initial Final
(Final ─ Initial)
1 20
2 20
3 20
5.3.5 Calculations
Standardisation of sodium thiosulphate solution
Molarity of K2Cr2O7 solution = M1 = mol dm−3
Volume of K2Cr2O7 solution = V1 = 20 cm3
Volume of Na2S2O3 solution used (from Table I) = V2 = cm3
Molarity of Na2S2O3 solution = M2 = ?

78
6M1V1 = M2V2
6M1V1
Molarity of Na2S2O3 solution = M 2 =
V2
= .. mol dm−3
Estimation of the strength of given Copper (II) solution
Molarity of Na2S2O3 solution = M3 = M2 = .......... mol dm−3
Volume of Na2S2O3 solution used = V3 = ......... cm3
Volume of copper (II) solution taken = V4 = 20 cm3
Molarity of copper (II) solution = M4 = ?
Using molarity equation M4 V4 = M3 V3
M 3V3 = ……….. mol dm-3

Molarity of copper (II) solution = M 4 =
V4
The strength of the given Cu (II) solution =
Molarity of the solution × atomic mass of copper = M 4 × 63 .5 = .......... g dm−3
5.3.6 Results
The strength of Cu (II) ions in the given solution is found to be = .g dm−3
The above value can be compared with the actual one which you can get from your
counsellor.
5.4 ANSWERS
1. i) l2 is almost insoluble in water.
ii) l2 is volatile in nature and is lost from an open container in a short
period. It requires standardisation every few days.
2. 2 CuSO 4 + 4 KI → 2 CuI+ 2 K 2SO 4 + I2

I2 + 2 Na2S 2O3 → 2 NaI+ Na2S 4O6
2 CuSO 4 + 4 KI+ 2 Na 2S 2O 3 → 2 CuI+ 2 K 2SO 4 + 2 NaI+ Na 2S 4 O 6
3. The iodine-starch complex is only slightly dissociated and a diffuse end

point will result if large amount of iodine were absorbed on starch.
4. Sodium bicarbonate produces CO2, in a solution containing KI, K2Cr2O7,

and acid and displaces the air present in it. Air present in the solution,
otherwise, would oxidise iodide to iodine and cause an error in the
titration.
79
EXPERIMENT 6
DETECTION OF EXTRA
ELEMENTS (N, S, X) IN THE
ORGANIC COMPOUNDS
Structure
6.1 Introduction Introduction
6.2 Classical Qualitative Organic Procedure

Analysis for Identification of a
Observations
Pure Compound
Result
Physical Examination
6.4 Detection of Nitrogen, Sulphur
Elemental Analysis
and Halogens by Middleton's
6.3 Experiment: Detection of Method
Nitrogen, Sulphur and
6.5 Lab Notebook Preparation
Halogens by Lassaigne Sodium
Fusion Method 6.6 Answers
6.1 INTRODUCTION
So far in this course we have introduced you to simple laboratory technique of
volumetric analysis in which you have learnt how the different types of
titrations are carried out. These quantitative analyses are useful in the
determination of amount of constituents in mixtures. The compounds isolated
from an organic reaction mixture or from some natural source may be
unknown. Identification and characterization of the structures of unknown
substances constitute vital part of organic chemistry. Introduction into this area
is provided by a study of qualitative organic analysis, which is an essential part
of the training of an organic chemist.
In this experiment, first we shall give you an overview of the stepwise

procedures that may be followed to identify a pure unknown compound using
classical methods. Then, we shall give you details of experimental procedure
for detection of extra elements such as nitrogen, sulphur and halogens in the
80 given organic samples.
Experiment 6 Detection of Extra Elements (N, S, X) in the Organic Compounds
In recent years, the development of chromatographic methods of separation

and analysis by spectroscopic techniques have revolutionised the qualitative
organic analysis. The chromatographic methods of separation will be
discussed in consecutive experiments and spectroscopic methods will be
discussed at higher level.

describe the steps involved in the identification of unknown compounds

by classical qualitative organic analysis;
carry out the physical examination of organic compounds;
write chemical equations for various reactions taking place while

carrying out various tests for elemental analysis; and
perform experiments for detection of extra elements such as nitrogen,

sulphur and halogens in the given organic samples.
6.2 CLASSICAL QUALITATIVE ORGANIC

ANALYSIS FOR IDENTIFICATION OF A
PURE COMPOUND
The classical qualitative organic analysis consists of a series of steps that help
to establish the identity of the unknown compound. These steps are:
Step 1. Physical examination
Step 2. Elemental analysis to determine the presence of elements other

than carbon, hydrogen and oxygen
Step 3. Solubility test in water, dilute bases and dilute acids
Step 4. Determination of physical constants such as melting point and

boiling point
Step 5. Functional group analysis using classification tests
Step 6. Preparation of derivatives
Steps 1 to 4, are called preliminary tests. While analysing organic compounds,

we can follow these four steps in any order but before performing the
qualitative tests for functional groups. Our final step must always be the
preparation of one or more solid derivatives. For performing final step, you
need a good understanding of preparative laboratory techniques. These
techniques will be discussed in Semester II Laboratory course.
Preliminary tests give us very important clues about the presence or

absence of certain class of the compounds. In this experiment we shall
consider only the basic preliminary steps 1: physical examination and
2: elemental analysis.
81
In an organic compound, elements carbon, hydrogen and oxygen are

assumed to be present commonly. In addition to these elements, they
may also contain nitrogen, sulphur and halogens (chlorine, bromine and
iodine). The elements nitrogen, sulphur and halogens are called as extra
elements.
The identification of extra elements in a given compound is a type of

qualitative analysis since the experiment is dealing with the composition of an
unknown compound. This experiment must be performed very carefully as
further analysis of the functional groups present in organic compounds is
according to the elements present in them. Experiments based on all the six
steps used for classical organic qualitative analysis will be performed in
Semester III Laboratory course. Let us discuss these basic preliminary steps
1and 2.
6.2.1 Physical Examination

In the physical examination, we consider the following points:
Check the sample purity: In this lab course we are providing unknown
organic compounds in pure form so it is not necessary for you to check the
sample purity. Otherwise, first step of qualitative organic analysis is the
purity checked by boiling point or melting point determination or using
chromatographic methods.
Note the physical state: The physical state of the compound whether it is
solid or liquid should be indicated.
Note the colour: The colour is also informative. Common coloured

compounds include nitro and nitroso compounds (yellow), α-diketones
(yellow), quinines (yellow to red), azo compounds (yellow to red). Phenols and
amines are often brown to dark- purple because of traces of air oxidation
products.
Note the odour: The odour of many organic compounds is highly distinctive.
Caution: Do not taste an Amines are recognisable by their fishy odour, esters are often pleasantly
unknown compound. To
fragrant. Alcohols, ketones, aromatic hydrocarbons and aliphatic alkenes have
note the odour,
characteristic odours. Thiols, isonitriles and low-molecular weight carboxylic
cautiously smell the
compound. Many organic acids possess unpleasant odours.
compounds are intensely
Make an ignition test: Take a small amount of given sample on a spatula
lachrymatory or worse.
and heat the spatula on a burner to see if the solid melts normally and then
burns.
Observe the flammability and nature of the flame. A yellow, sooty flame is
indicative of an aromatic or a highly unsaturated aliphatic compound, a yellow
but nonsooty flame is characteristic of aliphatic compounds. Halogenated or
highly oxygenated compounds often burn with difficulty or not at all (for
example carbon tetrachloride is used as fire-extinguisher).
The characteristic odour of sulphur dioxide indicates the presence of sulphur

in compound. Certain compounds like sugars char and leave a black residue
on the spatula and emit a characteristic odour.
82
If a white, non-volatile residue is left after ignition, add a drop of water and
treat the solution with litmus or pH paper. A sodium (or other metal) salt is
indicated by the colour change to basic colour.
Activity: Carry out Physical Examination and Ignition Test of Some

Organic Compounds
For this activity we are providing six organic compounds.
Requirements: Chemicals Apparatus

Samples Burner
Spatula
Procedure
Take six organic samples from your counsellor and follow the procedure as
mentioned above for physical state, colour, odour and ignition test for each
sample. Do not throw your samples after performing these tests; you will need
these samples for detection of extra elements.
Report your results in Table 1 given below:
Table 1: Physical examination of organic compounds
Sample Physical Colour Odour Ignition test Conclusion

No. state (Aromatic or Aliphatic)
1.
2.
3.
4.
5.
6.
Discuss your results with your counsellor.
SAQ 1
What kind of flame would you expect to see in an ignition test of the following
compounds?
a) Oxalic acid
b) Benzoic acid
c) D-Glucose
d) 1-Naphthol
6.2.2 Elemental Analysis

The technique of elemental analysis involves the determination of elements
which may be present in a compound. The halogens, sulphur, oxygen, and
nitrogen are the elements other than carbon and hydrogen that are most 83
commonly found in organic molecules. There is no direct method for detection

of oxygen. Its presence as part of a functional group becomes apparent later
when you perform tests for functional groups in the Semester III Laboratory
course. Presence of other hetero atoms may be detected using the Lassaigne
fusion technique where the organic compound is heated with metallic sodium,
or by Middleton's fusion technique using sodium carbonate and zinc in place
of sodium.
In both methods analysis is based on the conversion of the hetero atoms

present in the compound to water soluble inorganic salts such as cyanide,
Bonding in organic
sulphide or halide.
compounds is generally
covalent, therefore, like
1) Na or Zn/Na2CO3 +
inorganic compound, no C,N Na CN
−
direct method available 2) H2O

for the detection of
elements. In sodium 1) Na or Zn/Na2CO3 + 2−
fusion method covalent S Na 2S or ZnS
2) H2O
bonds of hetero atoms
are broken on heating of
organic compound with 1) Na or Zn/Na2CO3 + −
X Na X
sodium metal. This result 2) H2O
in the formation of
inorganic ions involving X may be Cl, Br, I
–
these elements: X ions
2–
from halogens, S ions
– Though, Middleton's method is less hazardous and also considered to be
from sulphide, CN ions
from nitrogen. These superior to Lassaigne fusion for the analysis of volatile compounds. But, for
ions can in turn be this method very pure zinc powder is required. In this course, we shall
readily identified by consider the Lassaigne fusion method first and then at the end of this
inorganic qualitative experiment, detailed method of preparation of aqueous solution for the
methods. elemental tests using Middleton's method will also be given. You can follow
any method depending upon the facilities available at your study centre.
6.3 EXPERIMENT: DETECTION OF NITROGEN,

SULPHUR AND HALOGENS BY
Owing to its potentially
hazardous nature, the LASSAIGNE SODIUM FUSION METHOD
fusion operation should
be carried out very For this experiment, we will be using same six compounds which you have
carefully. Keep your face used for carrying out physical examination.
away from the mouth of
the test tube at all times. 6.3.1 Introduction
Avoid pointing the fusion The basis of the sodium fusion procedure is as follows. On fusion with sodium
tube in the direction of the elements present in organic compounds are converted to ionic forms
anybody else. involving nitrogen, sulphur and halogens if they were present in the organic
compound. As mentioned above, nitrogen present in organic compounds gets
converted to cyanide ions, sulphur present in organic compounds to sulphide
ions and halogens present in organic compounds to halide ions. After the
organic compound has been heated with sodium metal, the residue is
hydrolysed with distilled water to destroy the excess sodium and dissolve the
inorganic ions that are formed as a result of fusion reactions.
84
1) Na + −
Na + C,N Na CN
2) H2O
Sodium cyanide
1) Na + 2−
Na + S Na 2S
2) H2O
Sodium sulphide
1) Na + −
Na + X Na X
2) H2O Sodium halide
X = Cl, Br or I
The aqueous solutions of these ions are divided into five portions and each
can be analysed by using following qualitative tests:
If, nitrogen and sulphur
i) Detection of Nitrogen are present in sample,
they will give precipitate
First portion of the aqueous solution is carefully acidified, followed by with silver nitrate.
addition of FeSO4, and then FeCl3. This converts the cyanide ions into
NaCN + AgNO3 → AgCN↓
ferric ferrocyanide, which precipitates as an intense blue solid called
+ NaNO3 (white ppt.)
‘Prussian blue’.
Na2S + AgNO3 → Ag2S
6NaCN + FeSO 4 Na 4[Fe(CN) 6] + Na 2SO4 + NaNO3 (black ppt.)
Sodiun ferrocyanide These precipitates may
Fe4[Fe(CN) 6]3 + 12NaCl interfere with the halogen
Na 4[Fe(CN) 6] + 4FeCl 3
test. Thus, it becomes
Ferriferrocyanide necessary to remove
(Prussain blue) cyanide and sulphide
ii) Detection of Sulphur ions from sodium fusion
extract before performing
In second portion of aqueous solution the sulphur can be detected by test for halogens.
precipitation as black lead sulphide with lead acetate solution and acetic
acid or by the appearance of violet or purple colour on addition of di-
sodium pentacyanonitrosylferrate (II) (sodium nitroprusside) in the third
portion.
Na 2S + Pb(CH3COO)2 PbS + 2CH 3COONa

Lead sulphide
(Black)
Na 2S + Na 2[Fe(CN) 5NO] Na 4[Fe(CN) 5NOS]

Sodium nitroprusside Sodium thionitroprusside
(Purple)
iii) Detection of Nitrogen and Sulphur when present together
When both nitrogen and sulphur are present together in an organic

compound, sodium thiocyanate is formed during sodium fusion. Their
presence can be detected by the appearance of blood red colour due to
the formation of complex ion thiocyanotoiron(III) on addition of ferric
chloride solution in the fourth portion of aqueous solution.
+ + -
NaCNS + FeCl 3 [Fe(CNS)]2 + 2Na + 3Cl
Sodium thiocyanate Blood red colour
85
iv) Detection of Halogens
Fifth portion of aqueous solution is acidified with dilute nitric acid and
boiled to remove any sulphide or cyanide ions that are expelled as
hydrogen sulphide or hydrogen cyanide, respectively. Sulphide and
cyanide must be removed because they interfere with the test for
halogens. Divide this portion in two parts. In the first portion add silver
nitrate solution. Presence of halogen in the compound is detected by the
formation of a precipitate of silver halide:
NaX + AgNO3 AgX + NaNO3
The colour of the precipitate provides a tentative indication of the halogen

present:
AgCl (White precipitate)
AgBr (Light yellow precipitate)
Agl (Dark yellow precipitate)
After completing the first step, add ammonium hydroxide to dissolve the
precipitate obtained on addition of silver nitrate. It is observed that the
white precipitate of silver chloride is soluble, the light yellow precipitate of
silver bromide is sparingly soluble and the dark yellow precipitate of silver
iodide is insoluble.
AgX + 2NH4OH [Ag(NH3)2]X + 2H2O
Positive identification of halide ions can be made by the following

inorganic qualitative test. Add carbon tetrachloride or chloroform, and
chlorine water to the second part of fifth portion of aqueous solution.
Shake the solution and check the colour of chloroform or carbon
tetrachloride layer (lower layer). Violet colour is due to iodide ions, orange
or brown colour is due to bromide ions and no colour and positive test with
AgNO3 indicate the presence of chloride ions.
Cl2
2NaBr 2NaCl + Br 2
Cl2
2NaI 2NaCl + I 2
SAQ 2
Discuss the role of sodium metal in fusion reaction.
SAQ 3
Why is it necessary to remove cyanide and sulphide ions from the sodium
fusion extract before testing for the presence of halide ions tests?
86
6.3.2 Requirements
Apparatus Chemicals
Apparatus for filtration (Fig. 6.1) Acetic acid
Burner Ammonium hydroxide
China dish Carbon tetrachloride or Chloroform
Five fusion tubes Chlorine water
Five test tubes Dilute hydrochloric acid (5 %)
Spatula Dilute nitric acid (5 %)
Test-tube stand Dilute sulphuric acid (5 %)
Tongs Iron(II) sulphate (Ferrous sulphate)
Tripod stand Iron(III) chloride, (Ferric chloride)

(5%).
Wire gauze
Lead acetate solution (0.15 M)
Ring clamp Samples of organic compounds
supported on stand
Silver nitrate solution (5%)
Sodium metal (4mm cube)/ Zinc dust

+ Sodium carbonate
Solution of disodium Caution: Manipulate

sodium with a knife and
pentacyanonitrosylferrate (II) (0.1%)
tongs or forceps. Never
(Sodium nitroprusside) touch it with fingers.
Wipe it free of kerosene
Fig. 6.1: Apparatus for filtration of
with dry filter paper.
fusion products
Red hot fusion tube will
shatter on contact with
6.3.3 Procedure water releasing any
unreacted sodium, and
Add 15 cm3 of distilled water into a clean china dish and place it near to your the wire gauze will stop
burner. Place about 20 mg of your sample in the bottom of a small fusion tube the loss of any material.
or in case of liquid take one or two drops of liquid in the fusion tube with the Do not throw any piece
help of a pipette or dropper. Use tongs to hold the fusion tube. Put a piece of of sodium into the sink. It
sodium roughly a 4 mm cube into the mouth of the test tube, without allowing may cause fire.
it to come into contact with the substance at the bottom. Heat the sodium The heating of aqueous
gently over a small flame until it melts and runs down into the sample. There solution with HNO3 has
the effect of removing
may be a very vigorous reaction when the molten sodium touches the sample.
any HCN or H2S, if
Heat the tube gently for one minute; and then heat more strongly until the cyanide or sulphides are
bottom of the tube glows red hot. Holding the wire gauze with tongs in your present which would
free hand, drop the red hot fusion tube into the water of china dish and cover it interfere with halogen
test. It may be omitted if
immediately with the wire gauze. If the fusion tube does not break, when it
nitrogen and sulphur
comes in contact with water, crush it with the help of a glass rod. Allow any have been shown to be
excess sodium to react. You may repeat the process of sodium fusion using absent.
one or more ignition tubes to obtain concentrated test solution. Place the
87
china dish on the wire gauze on a tripod stand and boil the solution for two
minutes (see Fig. 6.2). Filter the solution while hot, to remove the broken glass
and charred material and divide this aqueous solution in five equal portions in
five test tubes.
You may also perform China dish

Beilstein test for Wire gauze
detection of halogens.
Procedure is as follows:
Tripod stand
Heat the tip of a copper
wire in a burner flame
until there is no further Burner
coloration of the flame.
Let the wire cool slightly,
then dip it into the fusion
extract and again, heat it
in the flame. A green
flash is indicative of the
presence of halogen.
The Beilstein test is very Fig. 6.2: Arrangement for boiling sodium fusion extract
sensitive, thus halogen-
containing impurities 1. Detection of Nitrogen
may give misleading
results. This test is Add 200 mg of iron(II) sulphate to the first portion. Heat the solution to
unaffected by the boiling and add 2 drops of ferric chloride solution. Add sufficient dilute
presence of nitrogen.
sulphuric acid to dissolve any precipitate and make the solution acidic.
Cu + R–X → H2O + CO2 The formation of a deep blue precipitate or colouration (Prussian blue)
+ CuX
indicates the presence of nitrogen in the original organic compound. If the
Colour of flame is due to solution is green or blue-green, filter it; wash the filter paper with distilled
the formation of
CuX (X = Cl, Br, I) water, and examine the residue for blue colouration.
2. Detection of Sulphur
i) Acidify the second portion of aqueous solution with acetic acid, and
add a few drops of lead acetate solution. A black precipitate of lead
sulphide, PbS indicates the presence of sulphur in the original organic
compound.
ii) In the third portion of aqueous solution, add 2 cm3 of disodium

pentacyanonitrosylferrate (II) solution. The purple colouration which
fades slowly on standing confirms that sulphur is present.
3. Detection of Nitrogen and Sulphur together
Acidify the fourth aqueous portion with dilute hydrochloric acid. Add a few
drops of ferric chloride solution. Formation of blood red colour indicates
the presence of both nitrogen and sulphur together in the given
compound.
4. Detection of Halogens
To the fifth portion of aqueous solution, add sufficient nitric acid to render
the solution acidic and boil the mixture until its volume has been halved.
Divide this solution in two parts after cooling. In first part, add 1 cm3 of
silver nitrate solution. The observation of a white or yellowish thick
88 precipitate indicates the presence of halogen in the original organic
compound. A faint turbidity should not be interpreted as a positive test.

Tentative identification of the particular halogen may be made on the
basis of colour: Silver chloride is white, silver bromide is pale yellow, and
silver iodide is yellow. Now add 4-5 cm3 ammonium hydroxide to the
above precipitate and shake the contents. Observe the solubility of the
precipitate. Complete solubility of white precipitate, indicates the presence
of chlorine, a partial solubility of light yellow precipitate indicates the
presence of bromine and insolubility of dark yellow precipitate indicates
the presence of iodine in the organic compound.
To further confirm the presence of halogen, add 0.5 cm3 of carbon

tetrachloride or chloroform to the second part. Add chlorine water drop
wise to the mixture with gentle shaking. The appearance of a brown
colouration in the bottom layer indicates bromine, whereas a purple or
violet colouration indicates iodine. By the process of elimination, a sample
which gives a white precipitate with silver nitrate, but no colouration on
treatment with chlorine water must contain chlorine.
Repeat above mentioned procedure for all given six organic compounds and
report your results in Table 3.
6.3.4 Observations
Report your observations in detail for each sample in your practical note book
in the format given below:
Table 6.2: Identification of extra elements in the given organic sample
Sample No. Test Observation Inference
1. Test for Nitrogen:
................................................
Test for Sulphur:
..................................................
Test for Nitrogen and Sulphur

present together:
..................................................
Test for Halogens:
.................................................
6.3.5 Result
The given organic sample contains 1. ........, 2. ....... extra element(s).
Discuss your result with your counsellor.
Once you have completed experiments with these samples, try few more
samples which may contain one or more than one extra element.
89
6.4 DETECTION OF NITROGEN, SULPHUR

AND HALOGENS BY MIDDLETON'S
METHOD
In this method an organic compound is heated with a mixture of sodium
carbonate and zinc dust, nitrogen and halogens if present in the compound get
converted into water soluble sodium cyanide and halides, respectively and
sulphur gets converted into insoluble zinc sulphide. The water soluble cyanide
and halogens so formed are detected as discussed earlier in Lassaigne’s
method, whereas insoluble zinc sulphide is treated with acid and hydrogen
sulphide so evolved is detected by lead acetate.
Procedure
Add 15 cm3 of distilled water into a clean china dish and place it near to your
burner. Mix about 20 mg of the sample with a powdered mixture of zinc dust
(10 mg) and sodium carbonate (15 mg) and transfer this mixture in a small
fusion tube. In case of liquid take one or two drops of liquid in a fusion tube
with the help of a pipette or dropper and add the powdered mixture of zinc
dust and sodium carbonate. Use tongs to hold the fusion tube. Heat the fusion
tube first gently over a small flame and then strongly till it becomes red hot.
Holding the wire gauze with tongs in your free hand, drop the red hot fusion
tube into the water of china dish and cover it immediately with wire gauze. If
the fusion tube does not break when it comes in contact with water, crush it
with the help of a glass rod. Filter the solution whilst hot, preserve the residue
divide the filtrate in three parts and perform the test for nitrogen and halogens
as described in Lassainge’s method. To the residue left in the china dish, add
about 8 cm3 of dilute hydrochloric acid, immediately cover the china dish with a
filter paper moistened with lead acetate solution. Sulphur if present, in the form
of zinc sulphide will react with HCl to form hydrogen sulphide and will turn lead
acetate paper to brown or black due to formation of lead sulphide.
ZnS + 2HCl ZnCl 2 + H2S
H2S + Pb(OAc)2 PbS + 2AcH

Black coloured ppt
6.5 LABORATORY NOTE BOOK PREPARATION

In the beginning of this course, we have discussed some important points
which you should keep in mind while preparing a laboratory note book for
Chemistry experiments. Here we will discuss a possible format for organic
qualitative analysis experiments discussed above.
Each experiment should start on a fresh page, which should contain a title and
experiment number at the top. A sample note book format for the identification
of extra element(s) is given here.
Experiment No.....: Identification of extra element(s) in given organic sample

90
(sample No....).
1. Physical Examination:
Sample Physical state Colour Odour Ignition Inference

No. test (Aromatic or aliphatic)
1.
2. Elemental Analysis:
Sample Test Observation Inference

No.
1. Test for Nitrogen:

................................................
Test for Sulphur:
..................................................
Test for Nitrogen and Sulphur
present together:
..................................................
Test for Halogens:
.................................................
3. Result:
The given organic sample contains 1. ........, 2. ....... extra element(s).
6.6 ANSWERS
1. Observation Inference
a) Non-sooty flame Aliphatic Compound
b) Sooty flame Aromatic
c) Charring with smell of sugar Carbohydrate
d) Sooty flame Aromatic Compound
2. The organic compounds are basically of a chain of carbon atoms to which

various other atoms are covalently attached. Sodium is a strong reducing
agent that will cause the break of the carbon chain. It also will convert
those other atoms that are covalently bonded to the carbon chain to
inorganic ion. Note sodium itself is oxidized to Na+ in this process.
3. When we carry out AgNO3 test for halide ions, if cyanide and sulphide ion
are also present in sodium fusion extract, they will also precipitate as
AgCN and Ag2S along with precipitate of silver halide. Therefore, it
becomes necessary to remove CN- & S2- ions before performing AgNO3
test for halide ions. 91
BCHCL-132 Chemistry I
EXPERIMENT 7
SEPARATION AND
IDENTIFICATION OF THE
COMPONENTS OF A GIVEN
MIXTURE OF AMINO ACIDS BY
PAPER CHROMATOGRAPHY
Structure
7.1 Introduction Principle of Paper
Chromatography
Mechanism of Separation
7.2 Origins of Chromatography
Requirements
7.3 Terminology of
Chromatography Procedure
7.4 Experiment: Separation and Observations and Calculations

Identification of Amino acids in
Result and Discussion
a Mixture
7.5 Answers
7.1 INTRODUCTION
In chemistry, it is necessary to separate, isolate, purify and identify
components of complex mixtures. Simple separation techniques such as
distillation, crystallisation, extraction are quite useful when the components of
mixture are chemically and physically different and are few in number. For
example, the isolation of pure benzoic acid from its diethyl ether solution
requires the use of distillation to remove the more volatile diethyl ether. The
chemical structures, intermolecular forces and hence boiling points of these
two substances differ considerably, allowing the use of such a technique of
separation. However, where the components of a mixture resemble each
other very closely in structure, simple separation techniques are of little value.
For example, amino acids are closely related chemically and physically, none
of the techniques mentioned above, can be used for the separation of such
92 types of mixtures.
Experiment 7 Separation and Identification of the Components of a
given Mixture of Amino Acids by Paper Chromatography
Similarly when, we carry out reactions in the laboratory, many a times the
resulting product is not a single compound but a mixture having many
components which are structurally related. Thus techniques are needed which
could be used to separate such closely related substances. There are
techniques, known as chromatographic techniques available for separation
of such mixtures. In this experiment and in the next experiment, you will be
studying about these techniques. You will also perform two experiments:
Experiment 7 and Experiment 8 based on paper chromatography, which is
one of the simplest chromatographic technique.

explain the meaning of the terms chromatography, stationary phase and

mobile phase;
explain the mechanisms of partition as applied to chromatography;
calculate Rf values for different organic compounds;
set up experimental arrangement for paper chromatography; and
separate and identify amino acids from the mixture of two amino acids
by paper chromatography.
7.2 THE ORIGINS OF CHROMATOGRAPHY

The word chromatography is Greek in origin (chroma - colour; graph -
writing). The technique of chromatography was first applied to the separation
of coloured pigments and dyestuffs, as evident from the pioneering work of The term
Tswett who in 1906 separated the pigments from leaves into coloured bands chromatography is
by passing a solution of the pigments down a glass tube packed with applied to the process
powdered chalk. The separation of pigments that Tswett achieved was due to which involves the
distribution of a sample
the process called 'adsorption'. The most adsorbed pigment remains close to
between a stationary
the top of the column and the least adsorbed pigment is 'washed through' to phase and a mobile
form the lowest band in the column. phase.
In 1941 Martin and Synge used liquid-impregnated columns of absorbent to

obtain good separation of components in complex mixtures. The mechanism
A. J. P. Martin and
of separation was in this case a partitioning or distribution phenomenon
R. L. M. Synge were
involving each component in the mixture. We will consider the nature of these jointly awarded the Nobel
mechanisms of separation in more detail in the experiment part. Prize in 1952 for their
work on partition
Apart from column chromatography, many other chromatographic techniques chromatography.
were developed in last few decades. These can be listed as:
(1) Paper chromatography (PC)
(2) Thin-layer chromatography (TLC)
(3) Column chromatography (CC)
(4) Gas chromatography (GC) 93

(5) Gas-liquid chromatography (GLC)
(6) High resolution gas chromatography (HRGC)
(7) High performance liquid chromatography (HPLC).
The last four methods are useful in achieving a high degree of separation or
resolution and often involve use of elaborate and expensive apparatus.
Chromatography is used to isolate, purify the components of complex

mixtures in the laboratory both for analytical and preparative purposes.
Chromatography is used in labs right from the simple techniques such as
paper chromatography to the sophisticated ones such as Gas
chromatography and HPLC which find applications in research labs and
industries.
In this experiment, you will be using the simplest laboratory chromatographic

method, paper chromatography "which is relatively cheap." Nevertheless, the
basic principles remain the same and with skillful usage surprisingly good
separations can be achieved. You will also do some experiments based on
this chromatography during practical sessions. Let us now study the language
of these techniques. But before that answer the following SAQ.
SAQ 1
Which of the following characteristics would you say must be present for a
mixture to be satisfactorily separated by column chromatography? (Tick the
correct choice).
a) Be coloured
b) Be volatile
c) Be related by similar chemical structure

d) Show a differential adsorption to the column.
7.3 TERMINOLOGY OF CHROMATOGRAPHY

Various terms are frequently used and they form the language of
chromatographic techniques. These are explained below:
(i) Chromatography
The historical origins of the word chromatography have already been

discussed earlier. All types of chromatography have certain basic features,
although from the design of the apparatus, this may not be apparent. The
basic methodology involved is as follows:
The mixture to be separated is added to a moving 'solvent' which may be

liquid or gas. This moving stream, now containing components, is passed over
or through a fixed medium, which is specially designed to separate the
individual constituents in the original mixture.
94
However, there are many variations and hence in many cases, it is difficult to
see the common link between the various chromatographic methods.
(ii) Mobile Phase and Stationary Phase
The moving component in the chromatographic technique is called the mobile

phase, which is normally a liquid or a mixture of liquids, except in gas
chromatography where a gas is employed. The fixed medium is called the
stationary phase. Thus if the sample consisting of a mixture of components
(usually referred to as solutes) is added to the mobile phase, then the
components by a variety of physical processes will be carried along in the
moving stream to varying degrees.
Table 7.1 classifies some of the chromatographic methods. However, the

nature of the process whereby separation takes place is rarely confined to one
physical mechanism, and although the table indicates the nature of the
distribution process in each case, this is merely the most important factor.
Table 7.1: Classification of chromatographic methods
Nature of the Mobile Stationary Kind of Chromatography

Separation Phase Phase
Process
Partition Liquid Liquid Partition chromatography
Paper chromatography
Column chromatography
Partition Gas Liquid Gas-liquid chromatography
Adsorption Liquid Solid Adsorption chromatography
Thin-layer chromatography
Column chromatography
Ion-exchange chromatography
Adsorption Gas Solid Gas-solid chromatography
Now work through the following SAQ.
SAQ 2
For the examples given below, indicate what are (i) the mobile phase and (ii)
the stationary phase?
a) Paper chromatography
(i) (ii)
b) Thin layer chromatography
(i) (ii)
c) Partition column chromatography
(i) (ii)
d) Ion-exchange chromatography.
(i) (ii) ....
95
Let us now understand the basic ideas involved in partition chromatography by

performing the experiment given in the next Section.
7.4 EXPERIMENT: SEPARATION AND

IDENTIFICATION OF AMINO ACIDS IN A
MIXTURE
In this experiment you are going to separate and identify amino acids using
paper chromatographic technique. This chromatographic technique is the
simplest among all the chromatographic methods. It does not require much
complicated apparatus and surprisingly good separation can be achieved. Let
us now study the basic principle and mechanism of separation of this
technique.
7.4.1 Principle of Paper Chromatography

Paper chromatography is based on partition principle. The underlying principle
governing the separation of components in paper chromatography (liquid-
liquid) chromatography can be demonstrated using a static partitioning model.
Suppose a solvent, such as water, containing two solutes is added to a

separating funnel as shown in Fig. 7.1 (a).
(a)(a) (b)
(b) (c)
(c)
Fig. 7.1: Partial separation of solutes using two immiscible solvents
In Fig.7.1, the two solutes are represented by black and white circles. If a
second solvent immiscible with the first is added and the two solvents are
shaken together, then some of the solute molecules will transfer to the second
solvent forming the upper layer.
As you can see in Fig. 7.1 (b), the black molecules are more soluble in the
second solvent. If the stopper is removed and the first solvent is run off via the
tap, then a partial separation will occur, with white molecules predominantly in
the first solvent and black molecules predominantly in the second solvent.
This is shown in Fig. 7.1 (c).
You can show this phenomenon readily in your own laboratory as follows:
96
(1) Dissolve a small crystal of iodine in 5 cm3 of aqueous potassium iodide

solution in a test-tube. Note the brown coloration that forms.
(2) Then carefully add 5 cm3 of tetra chloromethane (carbon tetrachloride),

using a dropping pipette and shake the test-tube gently. Notice how the
tetra chloromethane layer, which was clear, is now purple.
(3) Cork the test-tube and shake vigorously, with care. The purple colour
in the organic layer intensifies while the aqueous layer becomes paler.
The colour changes that you observed in this experiment reflect the fact that Although solid iodine is
the iodine, which started off in the aqueous layer, has transferred mainly to the deep purple in colour, its
organic layer. No matter how much the mixture is shaken, the ratio of colour in KI solution is
concentrations of iodine in the two layers will remain constant, irrespective of brown while in the
organic layer it reverts to
the actual volumes of both solvents, provided only that the temperature
purple colour again.
remains constant.
When equilibrium is reached, i.e. there are no more tendencies for the iodine
to pass into the organic layer; we can define mathematically this partition or
Although it was originally
distribution process by the following expression: believed that the paper
chromatography is based
Concentration of solute in organic layer
= KD .(7.1) on partition principle.
Concentration of solute in aqueous layer Therefore it was
originally named as
Where KD is called the partition or distribution coefficient. paper partition
chromatography. It is
This value of KD is independent of the amount of solute taken or a volume of now generally
solvents used but depends on temperature because solubility is temperature recognised that the
dependent. paper more commonly
acts by a combination of
The explanation just given relates to static systems, where a component, on partition, adsorption, and
the basis of solubility consideration, partitions itself between two immiscible ion exchange
solvents. But in partition chromatography, one of the immiscible solvents is mechanisms.
mobile.
Let us now understand the mechanism of separation of components of solute

in paper chromatography.
To demonstrate the
7.4.2 Mechanism of Separation water content of a filter
paper, weigh a filter
In paper chromatography, the stationary phase is water held on paper in paper circle on an
cellulose fibers and the mobile phase is some solvent which runs up or down analytical or top-loading
balance (accurate to
the paper and the solutes are partitioned between it and the water. The
± 0.01 g). Dry the circle
solutes separate out according to their partition coefficients; the one which in an air oven set at 105
favours the mobile phase being carried along the paper more quickly. The °C for one hour. Then
components of solute are separated completely or partially in distinct colour cool it in a desiccator
zones if sample components are coloured, otherwise they are located by the and reweigh it. You can
application of different reagents. then calculate the
percentage of absorbed
To understand how the separation takes place, we need to look more closely water.
at the structure of the filter paper. We know that filter paper consists of
numerous cellulose fibers which attract a certain amount of water from the
atmosphere. Each fiber can be considered to be made up of a number of cells
and each cell consists of a fibrous part with its associated water. The 97
separation is achieved by partitioning the components of a mixture

between the moisture in the cells and the moving solvent flowing over these
cells. (Remember the static model which you have read earlier.) It is the
relative solubility of each component in the stationary phase (water) and the
mobile phase (solvent) which decides the rate at which the component is
transferred to the moving phase. Those components which are more
soluble in the mobile phase move further; and the components which are
more soluble in water will tend to remain longer in the water adhering to
the fibers.
A pictorial representation of the partitioning process is given in Fig. 7.2.
x
x
Moving solvent x x
x x
x x
x xx x
Hypothetical cell x
x
x x
x x
Cellulose fibre
Origin 1 2 3 Origin 1 2 3
(a) Start of separation (b) After one partition
x x x
x x
x x
Moving solvent x
x
x x
x
x x
x x x
x
Origin 1 2 Origin 1 2 3
3
(c) Solvent moves to the next (d) After second partition

cell carrying dissolved solutes
Fig. 7.2: A diagrammatic representation of the partitioning process in paper

chromatography
In this figure we have shown how the separation of two components has taken
place. Component A (represented by the circles) is twice as soluble in the
stationary phase, i.e. its partition coefficient is 0.5. Component B (represented
by the crosses) is twice as soluble in the mobile phase, i.e. its partition
coefficient = 2. Fig. 7.2 (a) represents the start of the process, where equal
amounts of A and B (equal numbers of crosses and circles) are dissolved
in the water cell. In Fig. 7.2 (b), an equal volume of moving solvent is in
contact with the first hypothetical water cell, and partitioning (i.e. dividing of
the components between the two liquids according to their solubility) will
take place so that twice the number of molecules of B will be found in the
mobile phase. Remember that B is twice as soluble in the moving solvent.
Pictorially, then, there are twice as many crosses as circles. The situation is
reversed in the static water cell where twice as many molecules of A are to
98 be found.
Chromatography is a dynamic partition process so that a mobile solvent

moves on to a situation represented by Fig. 7.2 (c). A second partition will take
place to give situation represented by Fig. 7.2 (d), where two adjustments will
take place - one at the origin and one at cell 1. At the origin, components A
and B will again partition into the mobile solvent in the ratio of 2:1 in favour of
B. Thus there are now two crosses in the moving phase and only one in the
water cell. A second readjustment will also occur between the mobile and
stationary phases in cell 1. In this case, however, as A is more soluble in the
water, there will be twice as many molecules of A in this phase (two circles as
opposed to one in the mobile phase).
By an extension of this process, it is possible to see why B is carried along in

the mobile phase to a greater extent than A. In practice, this process is
repeated countless times, eventually giving a sharp separation of the two
components in the form of two spots on a paper chromatogram (see Fig. 7.3).
The rate at which a component has moved is then determined by its
Retardation factor, (Rf) or Retention factor, which is defined as follows:
Distance moved by the centre of the solute spot

Rf =
Distance moved by solvent front
Thus, Rf values for component A will be as shown in Fig. 7.3.
D istance moved by the centre of spot of component A x A

Rf(A) = =
D istance moved by solvent front y
Similarly, retardation factor for component B, For good separation, the

components should have
D istance moved by the centre of spot of component B xB different Rf.
Rf(B) = =
D istance moved by solvent front y
Where, xA and xB are linear distances of centre of the spots measured from
the line of origin where mixture of solute is applied and y is the distance
travelled by solvent front from the starting point as illustrated in Fig. 7.3. Rf
values range from 0.00 to 1.00. Small Rf values indicate little tendency to
move with the solvent and thus reflect low solubility of solute in mobile phase.
Large Rf values conversely indicate a high solubility of solute in mobile phase.
So as long as the correct solvent and type of chromatography paper are used,
a component can be identified from its Retardation factor.
Solvent front
B
xA
y Rf(A) =
y
xB
A xB
Rf(B) =
Mixture xA y
Starting point
Fig. 7.3: Procedure for calculation of retardation factor (ratio), Rf 99

Solvent System (Mobile Phase) for Amino Acid Separation:
As mentioned above, the separation of the components mainly depends on

the relative solubility of the components of solute (sample) in mobile phase.
Therefore, the nature of solvent system plays important role in the
development of paper chromatogram. Various solvent systems have been
tried out for the separation of amino acids. While selecting a solvent system
for the separation of amino acids, some basic understanding of the structure
of amino acids is necessary.
As the name suggests, each amino acid contains an amino group, – NH2, and
a carboxylic acid group, – COOH. The molecular structure of a generic amino
acid is provided below:
H
(Amino group) H2N C COOH (Carboxylic group)
R
(Side chain)
Amino acids with nonpolar R Amino acids with polar R Amino acids with polar charged R
R = -CH3, Alanine R = -CH2OH, Serine R = -CH2COOH, Aspartic acid

R = -CH(CH3)CH3, Valine R = -CH2SH, Cysteine R = -CH2CH2COOH, Glutamic acid
R = -CH2CH(CH3)2, Leucine R = -CH2(OH)CH3, Threonine R = -(CH2)4 - NH2 , Lysine
R = -CH2Ph, Phenylalanine R = -(CH2)3 - NH(NH) - NH2, Arginine
There are 20 different amino acids that make up our proteins, and they closely
resemble in structure except the structure of the side chain R. In glycine, the
simplest amino acid, R is a hydrogen atom. Amino acids can be categorised
into three main types, on the basis of nature of their side chains. These are
amino acids with nonpolar, uncharged polar and charged polar side chains,
respectively. The nonpolar groups have generally an aliphatic or an aromatic
side chain. For examples, alanine, valine, phenylalanine, etc., have nonpolar
hydrocarbon side chains, therefore, are less soluble in water. The uncharged
polar types have side chains containing hydroxyl, amide or sulphydryl groups
(thiols). These amino acids like serine, cysteine and have polar but neutral R
groups thus tend to promote water solubility. The charged polar groups may
have carboxylic group (aspartic acid, glutamic acid) or a basic group (lysine,
arginine) in their side chain. Both acidic and basic R groups tend to promote
water solubility. Therefore, because of the differences in interaction of polar
and nonpolar side chains with water, amino acids will show different partition
coefficients i.e. relative solubility with solvent system of mobile phase and
water of stationary phase. Thus, one which favours the mobile phase being
carried along the paper more quickly and one which favours the stationary
phase remain near the base line.
There is no universal solvent system which is capable of separating all the

usual amino acids. We can try different solvent systems such as acidic
(phenol-water, 1-butanol-acetic acid-water), basic (pyridine-water,
1-propanol- aqueous ammonia) or neutral (1-butanol, methyl ethyl ketone)
solvent systems, which can provide us the best separation.
100
The paper chromatogram can be developed in many ways such as ascending:

here development of paper occurs due to the solvent movement in upward
direction on the paper; descending: here the development of paper occurs due
to solvent moves downwards on the paper; horizontal or radial mode: here the
solvent travels from centre towards periphery of circular chromatographic
paper; two dimensional: here the chromatogram development occurs in two
directions at right angle to each other. In our present experiment we will be
using ascending mode to develop our chromatogram.
For our present experiment, we will be following an easy ascending paper

chromatography technique and 1-butanol-glacial acetic acid-water in the ratio
12:3:5 as solvent system. This experimental set up does not require special
apparatus.
SAQ 3
In paper chromatography, the rate at which the components move in the
mobile phase is decided by which of the following factors:
a) The number of components in the mixture that is spotted on the paper
b) The relative solubility of the components in the mobile and the stationary
phases
c) The time for which the chromatogram is allowed to run
The basic principles discussed above now can be easily demonstrated

using the simplest of apparatus and chemicals. You can try the following
experiment at your Study Centre’s Chemistry Lab. The exercise will take up
very little time.
7.4.3 Requirements
Apparatus Chemicals
Chromatographic tank 1 1-Butanol
Boiling tubes 4 Glacial acetic acid
Measuring cylinder (100 cm3) 1 Any three amino acids from the
following:
Test tubes 5
[L-alanine, L-leucine, L-lysine
Spotting capillaries 5 L-aspartic acid, L-methionine]
Separatory funnel 1 Ninhydrin
Whatman No. 1 filter paper sheet
Spraying bottle 1
Solutions provided
1. Sample solutions: Provide solution of any three amino acids as

mentioned above. Make one unknown sample solution by mixing any two
of these three amino acids. 101
Their solutions can be prepared by dissolving 15 mg of each amino acid

separately in 1 cm3 of distilled water. Warm if a particular amino acid is
not soluble in cold.
2. Detector: Ninhydrin reagent (0.2%): Take 100 cm3 of 1-butanol and 100 cm3
of water in a separating funnel. Shake gently and allow it to form the layers.
Remove the lower aqueous layer. Transfer the upper organic layer to a
spraying bottle and to this add 0.2 g of ninhydrin, shake well and use as the
detector for amino acids.
7.4.4 Procedure
Proceed according to the following steps:
1. Preparation of Developer (solvent system): Prepare the developer as per

your requirement by mixing 1-butanol-glacial acetic acid-water in the ratio
12:3:5 by measuring the required volumes.
2. Place the developer in the chromatographic tank so that the height of the
developer is less than 1 cm. Place the lid and allow the tank to be
saturated with the solvent vapours.
3. Cut the chromatographic paper strip of the required size (usually 10 cm x

10-30 cm for four spot and breadth may be changed as per the number of
spot(s). On the strip draw a line with pencil at about 1 cm from one end.
This will be the bottom of the chromatogram. Mark off equally spaced
points along this line. (They should be separated by about 2 cm). Your
samples will be applied to these spots.
4. Apply the solutions of three known amino acids and solution of unknown
mixture of the amino acids to the point of application separately on the
marked strips using spotting capillaries or tooth picks. Use a fresh
capillary or tooth pick for each solution. Remember the position of the
known amino acids on the strip. You can label the paper at the top with
the name of each of the known amino acid and label the last as unknown.
5. After spotting, dry the spots by allowing the solvent to evaporate. You can
use a hot air dryer to dry the paper as illustrated in Fig. 7.4. Repeat
spotting process again.
(a) (b)
Fig. 7.4: Illustration of (a) spotting of solute sample using glass capillary tube
and (b) drying process using hair dryer
6. Suspend the spotted and dried paper strip in the chromatographic tank
with the help of a supporting glass road or hanger so that the lower end
touching the developer. Care should be taken to see that this is done
gently and the strip is vertical. The spots should always be above the
102 developer level (see Fig. 7.5). Cover the chromatographic jar with its lid.
Lid
Hanger assembly
Paper
Salple spot
Developer
Fig. 7.5: Some typical experimental arrangement for developing paper

chromatogram
7. Allow the developer to rise along the paper and wait till the developer
(solvent front) reaches near the upper end of the paper.
8. Remove the paper strip from the tank and mark the solvent front with the
help of a pencil.
9. Dry the strip until the acetic acid odour from the strip is no more present.
10. After the chromatogram has dried, spray the paper with the ninhyrin
reagent solution using a spraying bottle.
11. Heat the strip at 105 oC either in an oven or a hot plate or use hot air dryer
until the coloured zones of amino acids are seen.
12. Encircle the coloured zones and mark the centre of each zone.
13. Measure the distance of each spot centre from the starting line and also
the distance by which the solvent front has moved.
14. Calculate the Rf value for each spot on the strip. From the comparison of
Rf values, you can determine which amino acids are present in your
unknown sample. Report your results in the Observation Table I.
Alternatively you can run a paper chromatogram in 500 ml beaker by making a

paper cylinder as shown in Fig. 7.6. Common boiling tubes can also be used
to run paper chromatograms (see Fig. 7.6).
Cork
Beaker
Pin
Paper cylinder
Boiling tube
Paper strip
Staples
Spot Starting line
Staples
Developer solvent
Developer solvent
Boiling tube for developing Running a chromatogram by making paper cylinder
Beaker can be covered by aluminium foil
Fig. 7.6: An alternate method for developing paper chromatogram 103

7.4.5 Observations and Calculations

Observe the colour of the spots of various amino acids on paper
chromatogram. Measure the distance (x), to which the centre of spot for each
amino acid has moved from the original point of application and the distance
(y), to which the solvent front has moved from the original line on the paper.
Calculate the Rf values by the relation: Rf=x/y, for each known amino acid and
for the unknown. Present the data in an Observation Table below.
Observation Table I
xy
Amino acid x y
Rf =
1. --------
2. --------
3.---------
Given Mixture
Unknown A
Unknown B
Unknown A’s Rf resembles with ......
7.4.6 Result and Discussion

The given mixture sample contains:
1. A....................
2. B....................
The mobile phase rises up along the paper by capillary action, rapidly at first,
and then more slowly as the solvent front rises. The movement of an amino
acid along the paper depends on its solubility in mobile phase, and a number
of other factors. Therefore, the different amino acids move along the paper at
different rates and may have different Rf values. The significant differences in
Rf values of certain amino acids result into a clean separation. The most
widely used reagent for detecting amino acids is ninhydrin. Ninhydrin is the
2-hydrate of indane-1,2,3 trione (or triketohydrindene hydrate) with the
following formula.
O O
OH - H 2O
O
OH + H2O
O O
Hydrated form of Ninhydrin Unhydrated form of Ninhydrin
It reacts with amino acids to yield highly coloured products.
Heat
Amino acid + Ninhydrin coloured product
104
The different steps in colour formation between ninhyrine and amino acids are
shown below:
O O
H COOH
OH + - - 2H2O
H 3N COO N C H
OH R R
O O
- CO 2
Ninhydrin
- RCO
OH O
O
Ninhydrin
NH2
N - H 2O
O O
O
Purple coloured product
The formation of visible colour with ninhydrin has limits of detection that may
vary from 0.01-0.5 µg depending on the particular amino acid.
7.5 ANSWERS
1. You should have answered (d) as this is the chief mechanism of
separation on a column. It usually happens that there is a structural
similarity between components separated in this way though this is not an
essential characteristic.
2. a) (i) liquid – (ii) liquid b) (i) liquid – (ii) solid c) (i) liquid – (ii) liquid
d) (i) liquid – (ii) solid
3. (b) is the correct answer. Paper chromatography separations are mainly

decided by the differential solubility of the components between the two
liquid phases.
105
EXPERIMENT 8
SEPARATION AND
IDENTIFICATION OF THE
SUGARS PRESENT IN THE
GIVEN MIXTURE BY PAPER
CHROMATOGRAPHY
Structure
8.1 Introduction 8.4 Procedure
Expected Learning Outcomes 8.5 Observations and

Calculations
8.2 Principle
8.6 Result and Discussion
8.3 Requirements
8.1 INTRODUCTION
In the previous experiment you have separated amino acids by paper
chromatography. In this experiment you will again use paper chromatography
for the separation of sugars.
The term sugar applies to monosaccharides, and disaccharides, which are all
soluble in water and thereby distinguishable from polysaccharides. Many
natural sugars are sweet; however, the sweetness varies greatly with
stereochemical configuration.
Paper chromatographic method has proved to be of great significance for both

qualitative and quantitative separations of sugars. On a large scale, it has
proved to be of immense value in the sugar industries, in the analysis of fruit
juice and in a number of other fields.
Various developers may be used for particular paper chromatographic

separations. The developers which are used for this experiment may be
modified in order to achieve more satisfactory separations of certain sugar
mixtures.
106
Experiment 8 Separation and Identification of the Sugars present
in the given Mixture by Paper Chromatography

After studying this unit, you should be able to:
explain the principle of paper chromatography; and
separate and identify sugars by paper chromatography
8.2 PRINCIPLE
The basic principle of paper chromatography has already been discussed in
earlier experiment. In case of sugars, distribution of solute (sugars) between
the stationary and mobile phases, that is, the partition process is the major
factor in the paper chromatographic separation of sugars. Their partition
coefficients are substantially in favour of the aqueous phase. Therefore, with
non-aqueous developers, sugars appear on the paper chromatogram with low
Rf values, whereas with developers containing larger aqueous ratio, the Rf
values of sugars are much higher. This is because a sugar molecule
containing larger number of hydroxyl groups is readily soluble in water and
makes the partition coefficient in favour of the aqueous phase. Further, the Rf
values of sugars are affected by their structural formulae, their molecular
mass, the number of – OH groups, and presence of other kinds of groups such
as aldehydes or ketones etc. Commonly used mobile phase or developers for
separation of sugars are 1-Butanol-acetone-water (20:70:10), 1-butanol-acetic
acid -water (40:10:50), 1- Butanol-pyridine-water (60:40:30), etc.
In this experiment you will learn the paper chromatographic separation of simple
sugars.
8.3 REQUIREMENTS
Apparatus Chemicals
Chromatographic tank 1 Acetic acid, silver nitrate, ammonia
solution, anisidine hydrochloride
Boiling tubes 4
1-Butanol, sodium borate, phenol
Test tubes 5
red, methanol, aniline, acetone,
Measuring cylinder (100 cm3) 1 phthalic acid
Beaker (250 cm3) 1 Sugars (any three): D-glucose,
D-fructose, D-xylose, L-rhamnose,
Spotting capillaries 4
D-galactose, lactose, maltose,
Spraying-bottle 1 sucrose, D-mannose
Whatman No.1 filter paper sheets
Solutions Provided:
1. Unkown sugar sample solution: It can be prepared by dissolving any
two sugars in water from the list of three sugars given to students as
known sugars.
2. Detector: Any one or two of the following detectors may be prepared

considering the types of sugars given to students as samples. 107
Detector-1: Ammoniacal silver nitrate: Take 5 cm3 of saturated aqueous
solution of silver nitrate, add 50 cm3 of acetone, finally add ammonia
solution to make the solution clear and basic in nature.
Detector-2: Aniline hydrogen phthalate: Dissolve 1 cm3 of aniline and

1.66 g of phthalic acid in 100 cm3 of 1-butanol saturated with water. Both
these detectors 1 and 2 give coloured or dark spots with reducing sugars.
Detector-3: (For non-reducing sugars): Prepare a solution by mixing

0.25M sodium borate + phenol red + methanol in (1:2:7) proportion.
Detector 4: Aniline-Diphenylamine-phosphate: This reagent is superior to

other reagents for the location of sugars on the chromatogram. The
reagent can be prepared by mixing the following solutions before use in
the proportions stated below:−
(i) 4 percent solution of aniline in 95 percent alcohol (5 volumes);
(ii) 4 percent solution of diphenylamine in 95 percent alcohol

(5 volumes);
(iii) syrupy phosphoric acid (1 volume).
The reagent has the advantage of being effective for all reducing and non-
reducing sugars and oligosaccharides and is therefore, of general application
to all sugars. It produces blue, green, yellow or brown coloured bands with
sugars, the nature of the colour depending on the type of sugar.
8.4 PROCEDURE
For this experiment, you will be going to use same apparatus arrangement as
discussed for the earlier experiment.
Proceed according to the following steps:
1. Preparation of Solutions:
i) Developer (1-Butanol-acetone-water (20:70:10): using 100 cm3

measuring cylinder take 70 cm3 acetone, 20 cm3 1-butanol and 10 cm3
of water in a dry 250 cm3 beaker.
ii) Sample solutions: Prepare the aqueous solution of any three of the
following by dissolving 0.2-0.5 g of each sugar in 5 cm3 of water in a
small test tube. The sugars are: D-glucose, D-fructose, D-xylose, L-
rhamnose, D-galactose, lactose, maltose, sucrose, D-mannose (D-
glucose, D-fructose, D-xylose, L-rhamnose, D-galactose, D-mannose,
lactose, maltose are reducing sugars as they have aldehyde group in
open chain form).
2. Place the developer in the chromatographic tank so that the height of the
developer is less than 1 cm. Cover the tank with its lid and allow the tank
to be saturated with the solvent vapours.
3. Cut the chromatographic paper strip of the required size. On the strip
108 draw a line with pencil at about 1 cm from one end. This will be the bottom
of the chromatogram. Mark off equally spaced points along this line
separated from each other by about 2 cm. Your samples will be applied to
these points.
4. Apply the samples of three known sugar solutions and unknown sample of
mixture of sugar solutions to the points of application separately on the
marked strips using spotting capillaries or tooth picks. Use a fresh
capillary or tooth pick for each solution.
5. After spotting, dry the spots by allowing the solvent to evaporate. You can
use hot air dryer to dry the paper. Repeat spotting process again.
Remember the position of the known sugars on the strip. You can label
the paper at the top with the name of each of the known sugars and label
the last unknown mixture.
6. Suspend the spotted and dried paper strip in the chromatographic tank
with the help of a supporting glass road or hanger so that the lower end
touches the developer. Care should be taken to see that this is done
gently and the strip is vertical. The spots should always be above the
developer level (see Fig. 7.5). Cover the chromatographic tank with its lid.
7. Allow the developer to rise along the paper and wait till the developer
(solvent front) reaches near the upper end of the paper.
8. Remove the paper strip from the tank and mark the solvent front with the
help of a pencil.
9. Dry the strip until the odour from the strip is no more present.
10. After the chromatogram has dried, spray the paper with the provided
Detector reagent solution using a spraying bottle.
11. Heat the strip at 105 oC either in an oven or a hot plate or use hot air dryer
until the coloured zones of sugars are seen.
12. Encircle the coloured zones and mark the centre of each zone.
13. Measure the distance of each spot centre from the starting line and also
the distance by which the solvent front has moved.
14. Calculate the Rf value for each spot and also for the spots the mixture
contained. From the comparison of Rf values you can determine which
sugars are present in your unknown sample. Report your result in the
Observation Table.
8.5 OBSERVATIONS AND CALCULATIONS

Observe the colour of the spots of various sugars on paper chromatogram.
Measure the distance (x) to which the centre of spot of each sugar has moved
from the original point of application, and the distance (y) to which the solvent
front has moved from the original point on the paper.
Calculate the Rf values by the relation: Rf = x/y, for each known amino acid
and for the unknown. Present the data in an Observation Table below. 109
Observation Table
xy
Sugars x y
Rf =
1. --------
2. --------
3.---------
Given Mixture
Unknown A
Unknown B
8.6 RESULT AND DISCUSSION

Sugars present in the unknown sample are:
1. A .
2. B .
The colour of the sugar zones on the chromatogram depends on the detector
used. For example, detector-1 (ammoniacal silver nitrate) on reduction, it
results into metallic silver. Therefore, the reducing sugars give rise to brown-
black spots after heating to 100 °C.
O O
2Ag(NH 3)2OH + 2Ag + + H2O + 3NH3
− +
R H R O NH 4
Aldose sugars
The colour development with detectors 2, 3 and 4 depends on the following

mechanism. Heating of sugar with an acid produces furfural with pentose
sugar and 5 hydroxymethylfurfural with hexose sugars which can be
condensed with an aromatic amine or phenol to give characteristic colored
spots.
O OH
H OH O O
O
HO H HO H Heating HO H
or
H OH H OH −H2O
H OH H OH Detectors having amines

or phenols
OH OH give characteristic coloures
Glucose Fructose
110
FURTHER READING
1. Svehla, G. Vogel’s Qualitative Inorganic Analysis, Pearson Education,
2012.
2. Mendham, J. Vogel’s Quantitative Chemical Analysis, Pearson, 2009.
3. Vogel, A.I., Tatchell, A.R., Furnis, B.S., Hannaford, A.J. & Smith, P.W.G.,
Textbook of Practical Organic Chemistry, Prentice-Hall, 5th edition, 1996.
4. Mann, F.G. & Saunders, B.C. Practical Organic Chemistry Orient-

Longman, 1960.
111

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BCHCL-132

Prof. H. B. Singh (Retd.) School of Sciences,

Course Coordinator Prof. Bharat Inder Fozdar and

Expected Learning Outcomes

• explain the basic concepts involved in titrimetric analysis,

• explain the principles of acid-base, redox and iodometric titrations,

• take observations and calculate results after performing the experiments,

• separate and identify the mixture of compounds by paper chromatography.

Experimental Value − Correct Value

So, Best of Luck.

Expected Learning Outcomes

measure and deliver sample volumes by selecting and using appropriate

determine the mass of a sample by correctly using analytical balance;

perform basic laboratory skills, including pouring reagents and transferring

list and explain different types of titrations; and

explain the safety measures as applied to a chemistry laboratory.

1.2 APPARATUS COMMONLY USED

Pipettes which can 1.2.1 How to Use a Pipette

Another type of pipette is designed to deliver definite but different volumes of a

1.2.2 How to Use a Burette

Error in burette reading is among the most common sources of error in

10 Fig. 1.6: Delivery of liquid from a burette.

Fig. 1.7: Volumetric Flasks.

1.2.4 How to Use an Analytical Balance

Generally two types of analytical balances are used in chemical laboratories.

(i) Double pan analytical balance

(ii) Single pan electronic analytical balance

Fig 1.8: Analytical balance.

The zero point is + 0.25, i.e. 0.25 units to the right.

Fig. 1.10: Weight box and weights.

+ (Main division of the rider position × 0.001) g

+ (Subdivision of the rider position × 0.0002) g

Then the mass of the object

Mass of the substance = 15.00 g + (240 × 0.001) g + (2 × 0.001) g + (3 × 0.0002) g = 15.2426 g

1.2.5 Single-Pan Electronic Analytical Balance

Handling of Electronic Balance

Balances of all types should never be subjected to harsh treatment of any

14 Fig.1.11: Single-pan electronic analytical balance.

(1) weighing the container,

(2) adding its mass to the required mass of material, and

The relative molecular mass of oxalic acid dihydrate [(COOH)2.2H2O] crystals

The amount of a substance having mass equal to molar mass is called a

For titrimetric purposes we express concentration in terms of molarity

Number of moles of solute

The molarity, M, of a solution containing m g of the solute in V cm3 of a

Let the molar mass of the solute be Mm g mol−1

Number of moles of the solute

i.e., = mol dm−3 (1.1)

The molar mass of a substance is an inherent property. It is independent of

Percentage: The percentage of a solute in a given solution can be expressed

(c) In cases where the solute is also a liquid, it is possible to represent

Mathematically percentage is given as:

Formality: In certain ionic compounds, e.g., NaCl, which are completely

m1 = mass of the solute

Mm = Molar mass of the solute

The molality scale is useful for experiments, in which physical measurements,

1.4 STANDARD SOLUTION

A standard solution is defined as the one whose concentration (strength) is

Primary and Secondary Standards