IISER THIRUVANANTHAPURAM

School of Chemistry
CHY112

Semester I Chemistry Laboratory Manual

VARSHA-2023

Name:

Roll No:

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General Instructions

 Come to the lab in time and be prepared for the day's work
 Avoid loose talks and use of mobile phones in the laboratory.
 Always use laboratory coats and goggles. While handling hazardous chemicals
use masks and gloves.
 Do not smell chemicals by placing your nose at the bottle’s opening; waft the
smell toward your nose with your hand. Do not inhale vapors or make skin contact
with any substances.
 Never eat or drink in the lab.
 Waste should be disposed of in the appropriate containers:
 All accidents and dangerous occurrences must be reported immediately and staff
in charge of the laboratory must record all incidents in the accident book. An
emergency shower is located in the foyer area and also there are eyewash
stations. The first aid box is kept in the lab. You should ensure that you know a
general fire practice, whom to contact and where to go in an emergency.
 Students are expected to keep the laboratory and work bench clean. All students
should clean the apparatus and work bench after the work is over. Remove all
protective clothing and wash your hands before leaving the laboratory.
 All pre-lab work must be finished before the relevant laboratory session. Students
not completing the pre-laboratory task will be turned away from the laboratory
until the exercises are completed. Laboratory work in chemistry can be stimulating
to students who appreciate the challenge it offers to their abilities.
 Plan exercise, design procedure and perform experiments to improve the working
efficiency in groups.
 Good time management is the key to success in most areas of IISER life
 ----------------------------------------------------------------

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CONTENTS

Experiment 1: Basic Lab Techniques [2 Sessions]

a) Thin layer chromatography (TLC) and calculation of Rf values.

b) Determination of melting point of organic compound.
c) Column Chromatography: separation of organic mixture.

Experiment 2: Separation and quantification [1 session]

a) Separation of benzoic acid and naphthalene
b) Purification of organic compounds by crystallization
c) Determination of purity by melting points and TLC

Experiment 3: Isolation of Natural Products [1 Session]

Extraction of eugenol from cloves by steam distillation

Experiment 4: conversion of nitrobenzene to aniline [1 Session]

a) Reduction of nitro compound

Experiment 5: Titrimetric Estimations Based on Acid-Base Chemistry [1 Session]

(a) Standardisation of HCl solution using N/20 oxalic acid solution

(b) Estimation of alkali content in commercial antacid tablet.

Experiment 6: Redox-Titrimetric Estimations Based on Permanganometry [2

Session]

(a) Standardisation of potassium permanganate using sodium oxalate

(b) Preparation of K3[Fe(C2O4)3]3H2O

(c) Estimation of the oxalate content of Potassium trisoxalatoferrate(III) trihydrate

Experiment 7: Estimations Based on Iodometry [1 Session]

(a) Preparation and standardisation of sodium thiosulfate solution

(c) Solubility product of Ca(IO3)2 .

Experiment 8: Complexometric Estimations Based on EDTA [1 Session]

(a) Standardisation of EDTA solution using a standard zinc acetate solution

(b) Estimation of amount of calcium and magnesium in a milk sample.

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Experiment 1. Basic Lab Techniques

a) Thin layer chromatography (TLC) and calculation of Rf values

Thin-layer chromatography (TLC) is a sensitive, fast, simple, and inexpensive analytical
technique that will be used repeatedly in carrying out organic experiments.

The technique of TLC was useful in determining the type and number of ingredients in
the mixture, but it is not helpful for collecting the separated components. We could only
separate and visualize the spots.

In TLC, we use a stationary phase (most frequently silica gel) which is deposited over a
glass or aluminum support. We then can spot mixtures of compounds over the same
line. Then we elute the TLC with an organic solvent, and the different compounds will
move upwards at different rates, allowing the separation of the different components.
The stationary phase, silica gel contains Si–O–H bonds that bind to the different
compounds of the mixtures in a variable manner depending on the polarity of the
compounds. Also, depending on the nature of the solvent used (more polar or less polar),
it will pull upwards some compounds faster than others. In general, more polar
compounds will “climb” slower up through the TLC plate, and less polar ones will fly
upwards.

Rf = Retardation factor (or) Retention factor (or) Ratios of distance:

Retention factor (Rf) is the distance that a compound travels through the stationary
phase (TLC plate) between the origin spot and the distance the solvent front moved
above the origin.

To calculate the value of the Rf, you just have to apply this simple formula:

𝑚𝑜𝑣𝑒𝑑
Rf (spot) =

Please, keep in mind that retention factors depend greatly on the solvent system used
and on the stationary phase of the TLC. If you modify any of those, Rf will change. That’s
why when reporting retention factor values, it is essential to specify those parameters
for each compound. Finally, something that is very common while working with new
compounds: Many times the first choice of solvent system will not be the appropriate,
and just didn’t move from the base spot, or maybe they are somewhere between, but
still the separation is not perfect.

In any of these cases, you just have to keep tweaking the solvent system until you find
the most suitable for your compound! You generally want a solvent mixture that gives
both compounds a retention factor between 0.2 and 0.8.

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Procedure

Obtain a clean and dry silica gel TLC plate with the dimensions of 5 cm x 5 cm approx.
Draw a faint line in pencil 1.0 cm from the bottom of the plate. Be careful not to disturb
the silica gel as you draw the line! Under the line, mark lightly the name/no. of the
samples you will spot on the plate. Leave 1cm space between the samples so that they
do not run together.

Using capillary tube spot the samples on the marked point. When applying these
solutions to the plate, touch the capillary to the surface of the silica gel quickly and lightly
so the spot is very small. Place the TLC plate in UV chamber and observe the visibility
of spot. If the sample is too concentrated, it will visible as a smear and if it is not
concentrated enough, you will see nothing on the plate.

Prepare different eluting solvents (100% hexane to 30% ethyl acetate in hexane) and
pour the eluting solvent into the TLC chamber to a depth of approximately 0.5 cm. Place
the prepared TLC plates in the developing chamber. Cover it with the cover glass, and
allow the solvent front to move up the plate until it is approximately 0.5 cm from the top.
Do not disturb the TLC chamber while the chromatogram is developing! In this
case, the solvent will travel up the silica gel plate very quickly and will reach the top in
one to two minutes.

Remove the plate and mark the solvent front with a pencil. Allow the plate to dry for a
few minutes, and then observe it under short-wave ultra-violet light. With a pencil, circle
any spots that are illuminated. Outline the spots with a pencil and note anything
distinctive about any of the compounds.

Sketch a diagram of each chromatogram in your notebook. Measure the distance from
the origin to the center of each spot and the distance from the origin to the solvent front.
Calculate the values for each spot. Unknowns can be identified using these values.

Report

Suitable eluting solvent :

Rf Value of Sample A :

Rf Value of Sample B :

Rf Value of Sample X :

Unknown . …………………

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 b) Determination of melting point of organic compound
Melting point is a characteristic property of solid crystalline substance. It is the
temperature at which the solid phase changes to the liquid phase. This phenomenon
occurs when the substance is heated. Melting points are often used to characterize
organic and inorganic crystalline compounds and to ascertain their purity. Pure
substances melt at a sharp, highly-defined temperature (very small temperature range of
0.5 – 1 °C) whereas impure, contaminated substances generally exhibit a large melting
interval. The temperature at which all material of a contaminated substance is molten is
usually lower than that of a pure substance. This behavior is known as melting point
depression and can be used to obtain qualitative information about the purity of a
substance.

Procedure

Obtain a glass capillary melting point tube, which has one end sealed and the other end
open. The solid must be dry or the results will be affected as solvent can act as an impurity
and affect the melting range. If the solid is granular, pulverize the solid somewhat before
packing. Dip the open end of the capillary tube in the finely powdered naphthalene. Gently
tap the capillary tube on the table to fill the compound in the capillary tube to about a
length of 0.5 – 1cm. Insert the capillary tube containing the sample into melting point
apparatus and note down the melting point.

Report

Melting point of Organic compound =

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c) Column Chromatography: separation of organic mixture
Column chromatography is devised on the basis of differential adsorbance of
substances on solid adsorbent (silica or alumina) to an extent that depends on the
substance polarity and other chemical properties and structural properties. Some
compounds adsorb more strongly to the stationary phase than others, as they elute
(wash down) the column at a very slow rate. Thus they can be separated on the basis
of their elution rate accordingly. In this experiment, Column Chromatography will be
used to separate the compounds from mixtures and TLC will be used to monitor the
effectiveness of this separation.
Procedure

Take a clean dry column (30 x 300 mm) and plug in little amount of cotton. Fill the
column with Silica gel 60-120 mesh up to 10 cm height (for drying packing) or mix silica
with hexane (for wet packing). After that load the compound by dissolving it in 2-3 ml
DCM and put some little amount (up to 0.5 cm height) of sodium sulphate and lock the
column with little amount of cotton. Run the TLC with given mixture. Determine the
eluting solvent.

Run the column with 50mL of 5% ethyl acetate in hexane and collect the eluting solvent
in a dried 100 mL conical flask. Then increase the solvent polarity to 10% ethyl acetate
in hexane and continue to elute the column until TLC shows no more product is coming
down from the column. After collecting first compound increase the polarity to 50% ethyl
acetate in hexane (50 mL) and continue to elute the column until TLC shows no more
product is coming down from the column. Calculate the Rf value of separated
compounds.

Report

Rf value of component 1 =

Rf value of component 2 =

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Experiment 2: Separation, purification and quantification

a) Separation of benzoic acid and naphthalene by acid base

extractions
A technique called extraction will be demonstrated in this experiment. Extraction relies
on the solubility of substances into solvents and the immiscibility of the solvents into each
other. In this experiment, two organic compounds (benzoic acid and naphthalene) will
be separated from each other. These two are soluble in diethyl ether. By selectively and
sequentially reacting each organic compound, we can make it soluble in water and
insoluble in diethyl ether. Since di ethyl ether and water are immiscible in each other, they
will form two phases and can be separated from each other using a separating funnel.
The reacted organic compound which is in the aqueous portion is then converted back
into the original organic compound which precipitates out of the aqueous portion.

Step 1: Benzoic acid and naphthalene are soluble in ether. Sodium bicarbonate (aqueous)
will be added to the ether solution. Only the benzoic acid reacts with the sodium
bicarbonate.

Step 2: The aqueous layer is separated from the organic layer (extraction). The aqueous
layer contains the sodium salt of benzoic acid. The remaining ether layer is then
evaporated leaving naphthalene as white solid. The benzoic acid is then precipitated out
of the aqueous layer by acidifying it with HCl.

Procedure

Transfer the given mixture to a 150 mL Erlenmeyer flask. Add 100 mL of diethyl ether
(ethoxy ethane) and dissolve the sample by gently swirling. Pour the solution into a 250
mL separating funnel. Make sure that the stopcock is in the closed position. Rinse the
flask twice with 10mL of diethyl ether and transfer into the separating funnel. Add 25 mL
of 5% sodium bicarbonate (NaHCO3) to the separating funnel. This will react with the
benzoic acid to form a water-soluble salt. Stopper the separating funnel. While holding
the stopper in place with your fingers, invert the funnel. Point the spigot upward and away
from yourself and neighbors. Slowly open the stopcock to release any pressure in the
funnel. Close the stopcock and repeat the shaking - pressure release procedure until no
further pressure build up is noticed. This will indicate the benzoic acid/NaHCO3 reaction
is completed (no CO2 release).

Extraction of benzoic acid

Place the separating funnel into the ring stand to hold the funnel upright. Remove the
stopper from the funnel. Open the stopcock on the separating funnel and draw off the
lower aqueous portion of the liquid into a 125 mL Erlenmeyer flask. To this aqueous
portion carefully and slowly add 6 M HCl with constant stirring until a pH of 1-2 is

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indicated by pH paper. The benzoic acid will start to precipitate out as the pH is reduced.
Filter the benzoic acid using a Buchner funnel and use ice cold water to wash. Allow to
air dry.

Separation of Naphthalene

Transfer the remaining ether in the funnel to a 150 mL Erlenmeyer flask. Rinse the funnel
with an additional 10 mL of diethyl ether and combine. Add anhydrous sodium sulphate
(Na2SO4), about 1/10 the volume of the d i e t h y l ether solution to the 125 mL flask
containing the ether. Since there is about 1.5% water still dissolved in the ether, the
anhydrous sodium sulphate (Na2SO4), will absorb the remaining water. Allow the ethyl
ether solution to stand over the anhydrous sodium sulphate (Na2SO4), for 15-20 minutes
with occasional swirling. Decant the ethyl ether solution off the anhydrous sodium
sulphate (Na2SO4), into a dried 125 mL Erlenmeyer flask (Use a cotton plug in funnel if
necessary). Remove the diethyl ether by heating over water bath and obtained
naphthalene as white solid.

Purity of the compounds can be determined by either TLC or melting point.

Report

Rf value/ melting point of component 1 =

Rf value/ melting point of component 2 =

b) Purification of organic compounds by crystallization

The chemicals used for various purposes should be pure, completely free from any type
of impurities. Method of purification of a substance depends upon the nature of impurities
present in it. There are various methods for the purification of substances, e.g., filtration,
evaporation, decantation, distillation, and crystallization. Crystallisation is one of the very
important purification techniques, purifying substances by removing unwanted by-
products. The principle behind the crystallisation is that the amount of solute that can be
dissolved by a solvent increase with temperature.

In crystallisation, the impure substance is dissolved in a suitable solvent to reach its

nearly saturated solution at a temperature higher than the room temperature. At this high
temperature, the solute has very high solubility in that solvent, so a much smaller
quantity of hot solvent is needed for dissolving the solute than the solvent at room
temperature. When the solution is cooled, the pure substance is crystallised. The
solution left behind is called mother liquor. All the impurities are left behind in the mother
liquor. The purification method depends on the differences in solubility between the

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compound and the impurity. In this experiment we are recrystallizing the benzoic acid.
Benzoic acid is a colorless crystalline solid. It is highly soluble in hot water, but poorly
soluble in cold water. It can be recrystallized by dissolving it in hot water. The hot solution
obtained is filtered and cooled. Upon cooling, opaque white crystals of benzoic acid
crystallize.

Procedure.

Weigh 0.5 g of benzoic acid and transfer it into a 50 ml beaker. While constantly stirring,
gradually add a minimum quantity of boiling water just sufficient (~ 15ml) to dissolve the
benzoic acid. If required, heating can be used to dissolve the benzoic acid. Cool the
solution to room temperature by itself. Opaque white crystals of benzoic acid begin to
separate. Fix a fluted filter paper in a funnel and separate the crystals by filtration. Wash
the crystals with distilled water. Transfer the crystals to another filter paper and dry them
by pressing gently between the folds of the filter paper. Note down the recrystallized
benzoic acid and melting point of the recrystallized benzoic acid.

Report

Wt. of Recrystallized Benzoic Acid =

Melting point =

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Experiment 3: Isolation of Natural Products

Extraction of eugenol from cloves by steam distillation

Eugenol is widely used in dentistry, due to its analgesic, antiseptic balsamic qualities.
Cloves are dried flower buds of clove tree, Eugenia Caryophyllata, found in India and
other locations in Far East. Steam distillation of freshly ground cloves results in clove
oil. Clove oil belongs to large class of natural product called essential oils. The clove oil
consists of several compounds. Many compounds are used in as flavouring perfumes.
Eugenol is the major compound comprising 85-90%, eugenol acetate 9-10% and
caryophyllene. The structures are given below.

Steam distillation is used to isolate, separate or purify compounds that have high
boiling point or have low decomposition point. The compound to be distilled must be
insoluble or only slightly soluble in water. The clove oil and the water will co-distil.

Most compounds, regardless of their boiling point, will distil at temperatures below that
of boiling water. According to Dalton’s Law, Ptotal = P1 + P2. Boiling will occur when
the total pressure equals 760 torr. The vapour pressure of pure water is 100 oC at 760
torr. Since the vapour pressure of the second compound adds to the total pressure,
the mixture will distil at less than 100 oC. The boiling point of eugenol, an oil found in
cloves, is 248 °C, but it can be isolated at a lower temperature by performing a co-
distillation with water, this process is also known as a steam distillation. Since the
distillate will contain both water and eugenol, the eugenol must be extracted from the
water using an organic solvent. Once the eugenol is extracted into an organic solvent,
the organic layer is separated from the aqueous layer and dried. The eugenol is finally
isolated by evaporation of the organic solvent.

After obtaining the oil of cloves by steam distillation, you will need to isolate eugenol by
another useful technique known as chemically active extraction. In this process you
take advantage of the fact that eugenol, because it is a phenolic compound, is weakly
acidic whereas both eugenol acetate and caryophllene are neutral. Thus, treating the
clove oil with a strong base such as sodium hydroxide will result in the selective
formation of the sodium salt of eugenol which will be soluble in the aqueous phase.
The other neutral compounds will not react with sodium hydroxide and therefore will
remain in the organic phase. Simply by separating the aqueous phase and acidifying
it you can isolate eugenol.

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 Procedure

Steam Distillation:

Load a 100 mL round-bottomed flask with 5 g of freshly ground cloves, 50 mL of

deionized water, and a boiling chip. Add three drops of antifoam to the flask and set
up an apparatus for steam distillation as instructed in lab. Collect about 25 mL of
distillate in a graduated cylinder. Pour the distillate into a 125 mL Erlenmeyer flask,
add 2 spatula of NaCl, and gently swirl to dissolve the salt. Cool the solution in an ice
bath for a few minutes and pour into a 125 mL separating funnel. Add 50 mL of
hexanes to the separating funnel in order to extract the clove oil from the aqueous
phase. Venting frequently, gently shake the funnel for about a minute. (Remember:
vent often to release the pressure form within the funnel). Drain the aqueous layer into
the original Erlenmeyer flask and discard. Leave the organic layer containing clove oil
in the separating funnel.

Chemically active extraction:

Add 25 mL of 5% aqueous sodium hydroxide to the organic layer in the separating

funnel. Shake the funnel for about a minute and be sure to vent periodically. Drain the
aqueous layer into a beaker. Shake the organic layer with another 10 mL of the sodium
hydroxide solution and drain the aqueous layer into the same beaker. Discard the
organic layer. Cool the aqueous layer in an ice-water bath and acidify with 6M aqueous
hydrochloric acid until pH 1-2 is indicated by pH paper. Return the acidified solution to
the separating funnel and extract with 20 mL of hexane. Shake the funnel for about a
minute and with frequent venting. Collect the upper organic layer into a clean and dry
100 mL Erlenmeyer flask/beaker. Add a small amount of anhydrous sodium sulphate
(till the sodium sulphate in the flask is free flowing) to the organic layer and let stand
for 5 minutes. Decant the organic layer into a clean, dry, and pre-weighed 100 mL
beaker through a funnel plugged with cotton. Remove all the solvent by slow
evaporation and weigh the flask again. Calculate the weight of eugenol, estimate the
percent yield.

Report

Weight of cloves taken = g

Weight if Eugenol Obtained =

Percentage of recovery of eugenol =

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Experiment 4: Conversion of nitrobenzene to aniline

Reduction of nitro compound

Primary amines are obtained when nitro compounds are reduced by tin and
concentrated hydrochloric acid. Theoretically 1.5 mole of tin is needed for reduction of
nitro group and tin is oxidized to tin (IV) state. When reduction is complete, ammonium
chlorostannate salt may separate from which amine is liberated by basification, using
enough alkali.

Procedure

Into a 100 mL round bottomed flask equipped with a reflux condenser place 5 mL of
nitrobenzene and 8 g of granulated tin. Measure 25 mL of concentrated hydrochloric
acid. Pour about 10 mL of the acid down through the condenser and stir the contents
of the flask steadily. The reaction mixture becomes warm. If it boils vigorously
moderate the reaction by dipping the flask in tap water. When reaction has subsided
add other 10 mL portion of the acid and stir it steadily to ensure enough mixing and cool
again if reduction is very violent. Cool only sufficiently to keep the reaction under
control. Proceed in this way until 25 mL of the acid has been added. Finally heat the
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reaction mixture on a boiling water bath for 20-30 minutes with stirring using a magnetic
stirrer, until the solution become clear. (Reaction mixture become clear orange yellow
solution) Cool the reaction mixture to the room temperature (slow cooling, In air).

In a separate beaker (100 mL), add 17 g of sodium hydroxide into 25 mL water and
dissolve. Slowly add the sodium hydroxide solution drop wise to the above round
bottomed flask (which is cooled in a cold water bath) through the condenser as it is an
exothermic reaction. Then, stir the mixture until it reaches the room temperature.
Add about 1.5 g (2 spatula) of salt to salt out aniline. Transfer the reaction mixture in
to a 250 mL beaker and add 60 ml of distilled water. The hydroxide of tin which
precipitated first must dissolve, and solution should be strongly alkaline for allowing
aniline to be separate as oil. Add 40 ml of diethyl ether to the beaker and stir well. Aniline
dissolves in diethyl ether, decant the mixture in to a separating funnel (do not transfer
the solid lump settled in the bottom of the beaker).

Keep the flask to separate layers and drain down the aqueous layer into a 250 mL
beaker and the ether layer into a 100 mL beaker. Transfer the aqueous layer into the
separating funnel and add 20 mL diethyl ether, shake well, allow separating the two
layers. Collect the ether layer in the same beaker and reject the aqueous layer. Add 2-
3 spatulas of sodium sulphate into the organic layer swirl the beaker gently and leave it
for 2-3 minutes. Transfer the organic layer into a previously weighed beaker (250 mL).
Keep the ether fraction over boiling water bath and evaporate off the ether. After
evaporating the solvent note the yield of aniline. Store this aniline for the estimation.

Report

Yield = …………………….. g

TLC verification =

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Experiment 5: Acidimetry and Alkalimetry

When the strength of an acid is determined with the help of a standard solution of base,
it is known as acidimetry. Similarly, when the strength of a base (alkali) is determined
with the help of a standard solution of an acid, it is known as alkalimetry. Both these
titrations involve neutralization of an alkali. In these titrations H+ ions of the acid combine
with OH– ions of the alkali to form unionized molecules of water.

HA + BOH → BA + H2O

or H+ + OH– → H2O

The end point in these titrations is determined by the use of organic dyes (Indicator)
which are either weak acids or weak bases. These change their colors within a limited
range of hydrogen ion concentrations, i.e., pH of the solution. Phenolphthalein is a
suitable indicator in the titrations of strong alkalis (free from carbonate) against strong
acids or weak acids, Methyl orange is used as an indicator in the titrations of strong acids
against strong and weak alkalis. No indicator gives correct results in the titrations of
weak acids against weak bases. Such titrations are performed by some other methods.

Precautions

 Always rinse the burette and the pipette with the solutions to be taken in them.
 Never rinse the conical flask with the experimental solutions.
 Remove the air gaps if any, from the burette
 Never forget to remove the funnel from the burette before noting the initial reading
of the burette.
 Lower end of the pipette should always remain dipped in the liquid while sucking
the liquid.

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Estimation of Alkali Content in Commercial Antacid

Antacid generally contains weakly basic substances such as Al(OH)3, Mg(OH)2, MgCO3
etc. Alkali content of the antacid is determined by dissolving a weighed quantity of an
antacid in measured excess of standard HCl solution followed by back titration of the
excess HCl with a standard NaOH solution.

Mg(OH)2 + 2HCl = MgCl2+2H2O

Al(OH)3 + 3HCl = AlCl3+3H2O

HCl ≡ ½ Mg(OH)2 ≡ 1/3 Al(OH)3 ≡ 1 equivalent.

∴ 1000 mL (N) HCl solution ≡ 58.32/2 = 29.16 g of Mg(OH)2 ≡ 78/3 = 26g of Al(OH)3.

a) Standardisation of sodium hydroxide

Procedure:
Weigh out accurately 0.06 - 0.07g of A.R. crystalline oxalic acid into a 100mL volumetric
flask. Dissolve and dilute up to the mark with distilled water and then mix uniformly.
Pipette out an aliquot of 20mL of the standard oxalic acid in a 250mL conical flask and
add one drop of phenolphthalein indicator. Titrate the solution with the NaOH solution
until a light pink colour appears.

Normality of NaOH solution =

a) Standardisation of hydrochloric acid

Pipette out an aliquot of 20mL of the HCl solution in a 250mL conical flask and add one
drop of phenolphthalein indicator. Titrate the solution with the standard NaOH solution
until a light pink colour appears. Repeat the titration until you get concordant values.

Normality of HCl solution =

b) Estimation of alkali content in antacid

Transfer the given antacid suspension to a 100mL volumetric flask using funnel and
glass rod. Make up it into 100 ml with distilled water. Pipette out 20 ml solution into a
250 mL conical flask. Pipette out 40 mL of the standard 0.2N HCl solution to the conical
flask and mix it well. Keep it for 15 minutes with occasional shaking. Add one drop of

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phenolphthalein indicator and titrate the solution with the standard NaOH solution until
a light pink colour appears. Repeat the titration until you get concordant values.

Calculation

Standardisation of HCl

Sl.No. Vol. of Burette reading Vol. of NaOH

Hydrochloric reacted with HCl
Initial Final
acid(ml) (ml)

1 20

2 20

3 20

Estimation of Antacid (Back titration)

Sl.No. Vol. of HCl (ml) Burette reading Vol. of NaOH

reacted with
Initial Final
unreacted HCl
(ml)

1 40

2 40

3 40

Volume of HCl reacted with 20ml antacid solution

1000mL (N) HCl solution ≡ 58.32/2 = 29.16g of Mg(OH)2≡ 78/3 = 26g of Al(OH)3.

Report

Amount of alkali content in the antacid Mg(OH)2 / Al(OH)3 =

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Experiment No. 6: Redox Titrimetric Estimations Based on
Permanganometry

These are titrations in which potassium permanganate is used as an oxidising agent in

acidic medium. The medium is maintained by the use of dilute sulphuric acid. The
potential equation, when potassium permanganate acts as an oxidising agent, is:

2KMnO4 + 3H2SO4 → K2SO4 + 2MnSO4 + 3H2O + 5[O]

or MnO4– + 8H+ + 5e- → Mn2+ + 4H2O (E0 = 1.51V)

From which it follows that equivalent weight is one-fifth of the formula weight, is 31.606
g. The standard potential in acid solution 𝐸 has been calculated to be 1.51 volts, hence
permanganate ion in acid solution is a strong oxidizing agent. . Potassium permanganate
acts as a self-indicator. Before the end point, the solution remains colourless, but after
the equivalence point only one extra drop of KMnO4 solution imparts pink colour, i.e.,
appearance of pink colour indicates the end point.

Sulphuric acid is the most suitable acid as it has no action on permanganate ion in dilute
solution. But hydrochloric acid give the reaction.

- - + 2Mn++ + 5Cl2 + 8H 2O
2MnO4 + 10 Cl + 16H

Some permanganate may be consumed in the formation of chlorine. With a small excess
of free acid, a very dilute solution, low temperature and slow titration with constant
shaking, the danger from this reaction cause is minimized.

Preparation of Potassium trisoxalatoferrate(III) trihydrate.

Weigh about 1g of Fe(NH4)2(SO4)2·6H2O in a 150-mL conical flask and note down the
weight. Dissolve it in 3mL distilled water and add 1-2 drops of 3M H2SO4. Add 10mL
0.5M H2C2O4 to this solution and heat to boiling while stirring constantly to prevent
bumping. Remove the conical flask from the heat and allow the yellow precipitate of
FeC2O4 to settle. Decant the supernatant liquid (pour the liquid away from the solid) and
wash the precipitate using 5mL of hot distilled water. Swirl the mixture and allow the
precipitate to settle; decant and repeat the washing once more.

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Add 4mL of 1M K2C2O4 to the flask containing the precipitate, stir and heat to 40°C.
While the temperature is at 40°C, immediately add 2mL of 6% H2O2 drop wise and stir
continuously. Periodically check the temperature of the solution and make sure that it is
at least 40°C (but not >50°C) during the addition of H2O2. (Some brown Fe(OH)3 may
precipitate at this time.)

Heat the resulting solution to boiling. Obtain 4mL of 0.5M H2C2O4 and add 1mL of it all
at once to the boiling solution. Stir continuously and add the last few mL drop wise while
maintaining the temperature near boiling. The solution should turn clear green. If some
brown residue remains, add an additional 1mL of H2C2O4 drop wise, again while the
solution is boiling, until the solution is clear green. However, if the residue that remains
is yellow, it is probably unreacted FeC2O4, and more H2O2 should be added carefully.

If the solution is cloudy, gravity filter it into a clean Erlenmeyer flask. If it is clear, no
filtration is necessary. Then, while swirling constantly, slowly add 3-4mL ethanol to the
solution. Allow to cool, while an ice bath is prepared. Immerse the bottom portion of the
flask in the ice bath and stir slowly until crystals begin to form. Stop stirring and allow the
solution to stand in the ice bath for 20 minutes. A good crop of crystals should have
formed before the solution is filtered.

Filter at the pump. Wash with 10ml 1:1 ethanol and dry. Transfer the crystals to a pre-
weighed, labeled dry sample vial and record the weight of the product. This product
should store for further analysis.

Report

Weight of K3 [Fe(C2O4)3]·3H2O =

Percent yield =

Estimation of the Oxalate Content of K3[Fe(C2O4)3]·3H2O

Standard solution of potassium permanganate is used to analyze the oxalate content of

the potassium trisoxalatoferrate(III)trihydrate, K3[Fe(C2O4)3]·3H2O. In acid solution,
potassium trisoxalatoferrate(III) provides Iron(III) and oxalate ions. Iron(III) is not
oxidized by permanganate ion so the oxalate can be titrated directly. However, oxalate
ions react only slowly with permanganate ions at room temperature so the solution must
19 | P a g e
be warmed to about 60 ºC in order to make the reaction fast enough to be useful in a
titration.

2MnO4- + 5C2O42- +16H+ = 2 Mn2++ 10CO2 + 8H2O

Standardisation of potassium permanganate

Weigh out accurately 0.63g A.R. crystalline oxalic acid into a 100mL volumetric flask,
dilute up to the mark with distilled water and then shake to form a uniform solution.
Pipette out an aliquot of 20mL standard oxalic acid solution in a 250mL conical flask.
Add 20mL 4N H2SO4, heat nearly to 70º- 80ºC and then titrate the solution with the 0.1N
KMnO4 solution in hot condition until a faint pink colour stable for 30 seconds is obtained.
Record the titre value.

Estimation of the oxalate content

Weigh accurately about 0.04 g of the potassiumtrisoxalatoferrate(III) complex. Boil the

sample with 15 mL of 2 N sulfuric acid in a conical flask. Allow the solution to cool to
about 60°C and titrate slowly with the standard potassium permanganate solution.
Continue titration until the warm solution retains a slight pink colouration for 30sec.
Record the titre value. Calculate the percentage by weight of oxalate in the complex,
compare this with the theoretical value and thus obtain the percentage purity of the
complex. (1 ml 1 N KMnO4 = 0.044 g of C2O42- )

Calculation

20 | P a g e
Standardisation of KMnO4

Burette reading Vol. of KMnO4

Sl. Vol. of oxalic
reacted with
No. acid(ml) Initial Final H2C2O4 (ml)

1 20

2 20

3 20

Estimation of Oxalate Content

Sample 1 Sample 2

Mass of complex taken

Volume of KMnO4 titrated

Volume of 0.1N KMnO4 equivalent to ……. normal

KMnO4

Mass of oxalate in the complex

Percentage of oxalate in the complex

Report

Weight of Oxalate in the complex:

Sample I =

Sample II =

21 | P a g e
EXPERIMENT 7: ESTIMATIONS BASED ON IODOMERY

The reduction of free iodine to iodide ions and oxidation of iodide ions to free iodine
occurs in iodimetric and iodometric titrations.

I2 +2e → 2I– (reduction)

2I– → I2 + 2e (oxidation)

These are divided into two types:

(a) Iodimetric titration: These are the titrations in which free iodine is used. As it is difficult
to prepare the solution of iodine (volatile and less soluble in water), it is dissolved in
potassium iodide solution.

KI +I2 → KI3

This solution is first standardised before use. Substances such as sulphite, thiosulphate,
arsenite, etc., are estimated using standard solution of I2.

(b) Iodometric titrations: In iodometric titrations, an oxidation agent is allowed to react in

neutral medium or in acidic medium, with excess potassium iodide to liberate free iodine.

KI + Oxidising agent → I2

Free iodine is titrated against a standard reducing agent usually with sodium
thiosulphate. Halogens, oxy halogens, dichromates, cupric ion, peroxides, etc., can be
estimated by this method.

I2 + 2Na2S2O3 → 2NaI+ Na2S4O6

2CuSO4 + 4 KI → Cu2I2 + 2K2SO4 + I2

K2Cr2O7+6KI + 7H2SO4 → Cr2(SO4)3+ 4K2SO4 + 7H2O + 3I2

In iodimetric and iodometric titrations, starch solution is used as an indicator. Starch

solution gives blue or violet colour with free iodine. At the end point the blue or violet
colour disappears when iodine is completely changed to iodide. [When starch is mixed
with iodine in water, an intensely colored starch-iodine complex is formed. Many of the
details of the reaction are still unknown. But it seems that the iodine (in the form of
𝐼 ions) gets stuck in the coils of beta amylose molecules (beta amylose is a soluble

22 | P a g e
starch). The starch forces the iodine atoms into a linear arrangement in the central
groove of the amylose coil. There is some transfer of charge between the starch and the
iodine. That changes the way electrons are confined, and so, spacing of the energy
levels. The iodine-starch complex has energy level spacings that are just so for
absorbing visible light, giving the complex its intense blue color. The complex is very
useful for indicating redox titrations that involve iodine because the color change is very
sharp].

3. 1. Determination of Solubility product of Ca(IO3)2

Solubility products are relevant when a sparingly soluble ionic compound releases ions
into solution. A solubility product is a specific form of an equilibrium constant. Solubility
products change with temperature, so the temperature at which a solubility product was
measured must always be quoted. Many ionic compounds are sparingly soluble in water.
When added to water, most of the compound remains as an insoluble salt, but some
ions are released into the water to form a dilute solution. Even compounds we would
describe as insoluble often dissolve to a slight extent.

A sparingly soluble salt in contact with water maintains an equilibrium between the solid
and its ions. In simple cases, where there are no common ions or competing equilibria,
the ion concentrations depend only on the equilibrium constant for the particular salt.
When we talk about solubility equilibria we always write the equilibrium with the solid on
the left. For example, calcium iodate.

Ca(IO3)2(s)→ Ca2+(aq) + 2 IO3–(aq)

The solubility product expression is

Ksp = [Ca2+][IO3–]2

If an analytical technique is used to determine the concentration of either the Ca2+ or

IO3– ions in a saturated solution, the solubility product constant of Ca(IO3)2 can be
calculated. In this experiment the concentration of IO3– ions will be determined through
titration with a standardized solution of thiosulfate ion (S2O32–) in the presence of iodide
ion (I–), using starch as an indicator near the end of the titration. Iodate ion reacts with
iodide ions to give I3– (triiodide ion) as the sole product containing iodine:

IO3–(aq) + 8 I–(aq) + 6 H+(aq) → 3 I3–(aq) + 3 H2O(ℓ) (1)

23 | P a g e
Triiodide reacts with S2O32– ions during the titration, according to

I3–(aq) + 2 S2O32–(aq) → 3 I–(aq) + S4O62–(aq) (2)

Combining the two reactions gives the titration reaction of

IO3–(aq) + 6 S2O32– (aq) + 6 H+(aq) → I–(aq) + 3 S4O62–(aq) + 3 H2O(ℓ) (3)

3. 1. 1.Preparation of Saturated Ca(IO3)2 Solution.

Using the markings on a 250-mL beaker, prepare Ca(IO3)2 by adding 50 mL of 0.2 M

KIO3 to 20 mL of 1 M Ca(NO3)2. Stir vigorously with a stirring rod. A white precipitate of
Ca(IO3)2 should form. Let the mixture settle for a few minutes. Collect the precipitate
after decanting off most of the solvent. Wash the precipitate 3 times with ~5 mL portions
of distilled water. Add ~100mL distilled water, stir thoroughly with a stirring rod and let
the mixture stand for at least 30 minutes, leaving the stirring rod in the beaker. You are
attempting to make a saturated solution of Ca (IO3)2. Gravity filter the solution into a
Erlenmeyer flask (rinse the flask with first few ml of fitrate). Do not wash the precipitate
on the filter paper.

3. 1. 2. Standardisation of Na2S2O3 solution.

Prepare 100mL 0.1 N potassium dichromate solution. Take 20mL dichromate solution
using a pipette into a 250ml iodine flask. Add 6mL concentrate hydrochloric acid slowly
whilst gently rotating flask in order to mix the liquids. Add 10mL of 10% KI. Stopper the
flask and mix the solutions well and allow to stand for 5 minutes. Rinse the stopper down
into the flask and dilute to 100mL using distilled water. Titrate the liberated iodine against
sodium thiosulphate solution from the burette constantly shaking so as to mix the
solutions thoroughly. When the solution attains a yellowish green colour add 2mL of
starch solution to get to blue colour; rinse down the sides of the flask. Continue the
titration drop wise, and swirling the liquid constantly, until the colour changes from
greenish blue to light green on the addition of just one drop. Repeat the titration and
calculate the normality of the sodium thiosulphate solution.

3. 1. 3. Estimation of solubility product

Rinse a 20-mL pipette with a small amount of the filtrate. Pipette out 20.0 mL of the
filtrate into a clean 250-mL Iodine flask. Add 20.0 mL of distilled water to the flask. Add
about 0.2 g of solid KI and dissolve. Add 20 drops of 2 M HCl to the flask, immediately
24 | P a g e
stopper the flask and swirl to obtain a homogeneous red brown solution. Titrate with
standardized S2O32– until the solution becomes light orange to yellow. Add 1ml of 1%
starch solution. Continue titrating until the blue solution just turns colorless. Record the
final volume of S2O32–. Perform the estimation in duplicate.

Calculation

Standardisation of sodiumthiosulphate

Sl. Vol. of K2Cr2O7 Burette reading Vol. of thio reacted

No. (ml) Initial Final with HCl (ml)

1 20

2 20

3 20

Estimation of solubility product

Sl. Vol. of Ca(IO3)2 Burette reading Vol. of thio reacted

No. (ml) Initial Final with HCl (ml)

1 20

2 20

3 20

Report

Solubility product of Ca (IO3)2. =

25 | P a g e
EXPERIMENT 8: COMPLEXOMETRIC ESTIMATIONS BASED ON
EDTA

A titration, in which an undissociated complex is formed at the equivalence point, is

called complexometric titrations. These titrations are superior to precipitation titrations
as there is no error due to co-precipitations. EDTA (ethylenediamine tetra-acetic acid) is
a useful reagent which forms complexes with metals. In the form of disodium salt, it is
used to estimate various metal ions in presence of eriochrome black-T or other suitable
metal ion indicator.

To simplify the discussion, ethylenediaminetetra-acetic acid is assigned by the formula

H4Y: and the sodium salt is Na2H2Y and affords the complex forming ion H2Y2- in the
aqueous solution. The usefulness of EDTA as the titrant is due the presence of six atoms
which are available for co-ordination to the metal ions in such a way that five membered
rings are formed. 1:1 complexes are usually formed and these are the most important
and the reactions with M++ cations may be written as

The ionization of the complex is dependent upon the 𝑝 of the solution. Lowering of the
pH decreases the stability of the metal-EDTA complex. The more stable the metal-EDTA
complex, the lower is the 𝑝 at which an EDTA titration is possible.

Let us consider the metal ion indicator which forms 1:1 complex. The use of metal ion
indicator in an EDTA titration may be written as

M – In + EDTA ⟶ M – EDTA + In

This reaction will proceed if the metal indicator complex is less stable than metal-EDTA
complex. During titration the metal ions are progressively complexed by EDTA until
ultimately the metal is displaced from the complex M – In to leave the free indicator (In).
26 | P a g e
Thus Eriochrome Black T, which may be written as H2In- exhibits the following acid base
behavior.

pH -- pH
- HIn ---
H 2In In
5.3- 7.3 10.5 - 12.5
Red Blue Yellow-orange

4. 2. Estimation of Calcium in Milk

4. 2. 1. Standardisation of EDTA

Prepare 0.01M zinc sulphate solution in 100ml volumetric flask. Pipette out 20mL of
standard zinc sulphate solution in a 250mL conical flask,dilute to 100mL with deionised
water, add 2mL of NH3-NH4Cl buffer solution and a pinch of eriochrome black T indicator
when the colour of the solution turns wine red. Titrate the solution with the EDTA solution
until wine red colour turns pure blue.

The last trace of a reddish shade should disappear at the end point. Titrate slowly near
the end point.

4. 2.2 Estimation of Calcium in Milk

Pipette out 20mL of the supplied milk solution in a 100mL beaker, precipitate proteins of
milk by trichloroacetic acid and filter into 100mL volumetric flask. Wash with deionised
water, dilute up to the mark with deionised water and mix uniformly. Pipette out 20mL of
the diluted supplied milk solution in a 250mL conical flask, dilute to 100mL with deionised
water, add 2mL of NH3-NH4Cl buffer solution and a pinch of Eriochrome Black-T indicator
when the colour of the solution turns wine red. Titrate the solution against the EDTA
solution until the wine red colour turns pure blue. No tinge of reddish blue should remain
at the equivalence point. Titrate slowly near the end point. (1mL (M) EDTA = 0.04008g
of Ca).

27 | P a g e
Standardisation of EDTA

Sl. Vol. of ZnSO4 Burette reading Vol. of EDTA

No.
(ml) Initial Final (ml)

1 20

2 20

3 20

Estimation of Ca2+

Sl. Vol. of Ca2+ Burette reading Vol. of EDTA

No.
(ml) Initial Final (ml)

1 20

2 20

3 20

Report

Percentage of calcium in the supplied milk =

28 | P a g e