LABORATORY MANUAL

FOR

CHEMISTRY LABORATORY CHEM F110

By
Chemistry Department

EDUCATIONAL DEVELOPMENT DIVISION

BITS Pilani, Dubai Campus
DIAC, Dubai, UAE
2022- 2023

1
 BITS Pilani, Dubai Campus
DUBAI INTERNATIONAL ACADEMIC CITY,
DUBAI, UAE.

CERTIFICATE

This is to certify that Mr. /Ms. ____________________________________

ID No.______________________________________ has successfully completed

all the experiments in the Chemistry Laboratory course (CHEM F110) during the

First Semester 2022-2023 and has submitted the report for the same.

Date Name and Signature of the Course Faculty

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 PREFACE

This manual is designed primarily to serve as an instrumental book for students,

technical staff, and instructors to assist in performing and understanding few
experiments in Chemistry. It is important that the students perform/carryout
experiments in the laboratory to understand the theories that they have studied in
the theory course. An attempt has been made to identify concepts that are interest
or challenge to students and can be better understood through laboratory work.
This will help the students to learn about Chemistry, using laboratory equipment
data interpretation and presentation. This manual contains ten experiments. This
laboratory manual specifically provides:

1. An introduction of important safety information.

2. Objective behind each experiment.
3. List of requirements.
4. Theory of the experiment.
5. Experimental procedure.
6. Tabulation for recording the readings.

In conclusion, we view this manual as one of the continual modifications and

improvement. We encourage discussing ideas for improvements and suggestions
for new experiments. For any new specific experiment HOD or instructor may be
contacted. This manual is the culmination of the efforts of the faculty members of
Department of Chemistry, BITS Pilani-Dubai Campus. Chemistry group gratefully
acknowledge the support and encouragement received from Director, Dr.
Srinivasan Madapusifor the preparation of this laboratory manual.
TABLE OF CONTENTS

Experiment Page
Title of the Experiment
No No
1 Preparation and Characterization of Aspirin 6

3
 (Acetylsalicylic acid)
Preparation of Co-ordination Complex Tetrammine
2 11
Copper (II) Sulphate Mono hydrate
3 Estimation of Ferrous Sulphate by Permanganometry 15

4 Determination of the Hardness of Water by EDTA. 20

5 Identification of Organic Compounds 27

6 Determination of Ionization Constant of a Weak Acid 39

7 Verification of Ostwald’s Dilution law 47

Determination of Solubility and Heat of Solution of
8 Sparingly Soluble Substance 52

9 Kinetics of Decoloration of Dye in Alkaline Condition 58

Colorimetric Estimation of Dye Concentration in Water
10 63
Sample
Demonstration
67
1 Estimation of Sodium by Flame photometry

INDEX

4
 TABLE OF CONTENTS

Experiment Signature of
Title of the Experiment
No the Faculty
Preparation and Characterization of Aspirin
1
(Acetylsalicylic acid)
Preparation of Co-ordination Complex
2
Tetrammine Copper (II) Sulphate Mono hydrate
Estimation of Ferrous Sulphate by
3
Permanganometry
Determination of the Hardness of Water by
4
EDTA.
5 Identification of Organic Compounds

Determination of Ionization Constant of a Weak

6
Acid
7 Verification of Ostwald’s Dilution law
Determination of Solubility and Heat of
8 Solution of Sparingly Soluble Substance

Kinetics of Decoloration of Dye in Alkaline

9
Condition
Colorimetric Estimation of Dye Concentration
10
in Water Sample
Demonstration Estimation of Sodium by Flame photometry

EXPERIMENT – 1

Preparation and Characterization of Aspirin (Acetylsalicylic acid)

Objective
5
1. To prepare Aspirin.

2. To determine its Melting Point

3. To provide exposure to medicinal chemistry.

Requirements
(a) Apparatus

100 ml Beaker, measuring cylinder 10 mL, Beaker 250 mL and 500 mL, Glass stirrer, Vacuum
pump, Filter paper, weighing balance, Hot Water Bath, Watch glass, Dropper, Melting Point
Apparatus

(b) Chemicals

Salicylic acid = 2g

Acetic anhydride = 4 mL

Conc. Sulfuric acid = 5 drops

Ice cold Water = 100 ml

Historical Background

One of the most widely used nonprescription drugs is aspirin. It is an effective analgesic (pain
killer) that can reduce the mild pain of headache, toothache, neuralgia (nerve pain), muscle pain,
and joint pain (from arthritis and rheumatism). Aspirin behaves as an antipyretic drug (it reduces
fever) and it is an anti-inflammatory agent capable of reducing swelling and redness associated
with inflammation. It is also an effective agent in preventing strokes and heart attacks due to its

ability to act as an anticoagulant by preventing platelet aggregation. The father of modern

medicine was Hippocrates, who lived sometime between 460 B.C and 377 B.C.

Hippocrates left historical records of pain relief treatments, including the use of powder
made from the bark and leaves of the willow tree to help heal headache, fever and pains. By

6
1829, scientists discovered that it was the compound salicin in willow plants that gave pain
relief. It was a short time later that a tiny amount of bitter tasting, yellow, needle-like crystals
were isolated from willow bark by Johann Buchner, Professor of pharmacy at the University of
Munich. By 1829, a French chemist improved the extraction procedure and was able to obtain
about 30g from 1.5 kg of bark. In 1838, an Italian chemist was able convert salicin to an acid of
crystallized, colorless needles, which he named salicylic acid. Since salicylic acid contains
phenolic and carboxylic acid groups, it was irritating to the mouth, throat, and stomach. After
considerable work, the derivative acetylsalicylic acid was found to have similar pain relieving
properties without the harsh side effects. It was in 1899, that the Bayer Company in Germany
patented the ester acetylsalicylic acid and marketed the product as “aspirin” in 1899. Their
studies showed that this material was less of an irritant and the acetylsalicylic acid hydrolyzed in
the small intestine to salicylic acid, which was then absorbed into the bloodstream. The
relationship between salicylic acid and aspirin is shown in Figure 1. The first person to do so was
a French chemist named Charles Frederic Gerhardt. In 1853, Gerhardt neutralized salicylic acid
by buffering it with sodium (sodium salicylate) and acetyl chloride, creating acetylsalicylic acid.
Gerhardt's product worked but he had no desire to market it and abandoned his discovery. In
1899, a German chemist named Felix Hoffmann (Fig. 2), who worked for a German company
called Bayer, rediscovered Gerhardt's formula.

Felix Hoffmann made some of the formula and gave it to his father who was suffering from the
pain of arthritis, with good results. Felix Hoffmann then convinced Bayer to market the new
wonder drug. Aspirin was patented on February 27, 1900. The folks at Bayer came up with the

7
name Aspirin, it comes from the 'A" in acetyl chloride, the "spir" in spiraeaulmaria (the plant
they derived the salicylic acid from) and the 'in' was a then familiar name ending for medicines.
Aspirin was first sold as a powder. In 1915, the first Aspirin tablets were made. Interestingly,
Aspirin® was once a trademark belonging to Bayer. After Germany lost World War I, Bayer was
forced to give it up as part of the Treaty of Versailles in 1919.

Procedure

1. Weigh approximately 2 g of salicylic acid using a watch glass and transfer the salicylic
acid completely to a clean, dry 100 ml beaker.

8
 2. In the hood, transfer 4.0 mL of acetic anhydride into the 100 ml beaker using a 10 mL
graduated cylinder and carefully add 5 drops of concentrated sulfuric acid to the beaker
using the dropper provided. Be careful not to let the sulfuric acid drop anywhere other
than into the beaker. Swirl the beaker to mix the contents. Do not add the sulfuric acid
before the acetic anhydride.
3. Keep the beaker in a boiling water bath. Make sure the reaction mixture is below the
liquid level in the bath and that the water is boiling. The reaction mixture should form a
clear solution as it is heated. Continue the heating for 20 minutes by which time all of the
solid should have dissolved.
4. After removing the beaker from the water bath, measure 10 mL of ice-cold water in a 10
mL graduated cylinder and add it to the reaction mixture, to hydrolyze the unreacted
acetic anhydride.
5. Chill the contents in the beaker in an ice bath with occasional swirling to cause
precipitation of the product. Add 25 mL of ice-cold water to the reaction mixture. If no
precipitate forms, scratching the side of the beaker with a glass stirring rod may help
initiate crystallization. Filter the product, dry, weigh and report.
6. Determine the melting point of it.
Calculations
Mass of salicylic acid used = ----------      g
 Molar mass of Salicylic acid = 138.121 g/mol
No. of moles of salicylic acid = Mass of salicylic acid used / Molar mass of Salicylic 
acid
=   ------------- moles
Density of Acetic anhydride = 1.08g/cm3
Volume of acetic anhydride used = ---------- ml
Mass of Acetic anhydride = Volume x Density = ------------- g
Molar mass of Acetic anhydride   = 102.09g/mol
No. of moles of acetic anhydride = Mass of Acetic anhydride used / Molar mass of
Acetic anhydride
No. of moles of acetic anhydride = ---------------- moles

9
The mole ratio of salicylic acid to acetic anhydride        = ----------: -----------
 Which is the limiting reactant Salicylic acid or acetic anhydride?  
No of moles of limiting reactant =   --------------moles
No moles of aspirin to be produced = ----------- moles 
Molecular mass of aspirin = 180.157 g/mol
Theoretical yield = Molecular mass of aspirin x No moles of aspirin to be produced 
                             = -------------- g
Mass of Aspirin produced        =     g
Percent yield      =   Mass of Aspirin (g) x 100/ Theoretical yield
                               =           %

Result Date:

Percent yield of Aspirin = _________

M. Pt. of Aspirin = ________

EXPERIMENT – 2

Preparation of Co-ordination Complex TetrammineCopper (II) Sulphate

Monohydrate
Objective:

1. To prepare a pure sample of Tetrammine Copper (II) Sulphatemonohydrate.

2. To provide experience in the synthesis of co-ordination compounds.

Requirements

(a)Apparatus

Vacuum Pump, Beaker 250 ml, Glass rod

(b)Chemicals

Copper sulphate solution (saturated) = 5 ml

10
 Ammonia (1: 1) solution = as required
Ethanol = 15 ml

Theory:

Coordination compounds (or) Complex compounds:

Coordination compounds retain their identities even when dissolved in water or any other
solvent and their properties are completely different from those of the constituent ions.

(E.g.): Potassiumferrocyanide is a complex compound. It is formed by adding KCN to a

saturated solution of ferrous cyanide.

Fe (CN) 2 + 4KCN → K4 [Fe (CN) 6]

When K4 [Fe (CN) 6] is dissolved in water, it does not give the usual tests for Fe 2+ and CN–
ions. This indicates that these ions which were originally present are not formed during
dissolution.

Actually, these ions are present in the form of a new ion called ferrocyanide ion which is a
complex ion and does not ionize into constituent ions.

K4 [Fe (CN) 6] →4K+ + [Fe (CN) 6]4—

The dissociation of ferrocyanide ion is so negligible, as not to confirm to the usual tests of the
ferrous ion.

The cation to which one or more neutral molecules or anions are attached is called the Centre of
coordination. Any atom, ion or molecule which is capable of donating a pair of electrons to the
central atom is called a coordinating group or ligand.

In a ligand the atom which actually donates the electron pair is called the donor atom. The
ligand containing one two or more donor atoms are known as unidentate, bidentate and
multidentate, respectively.

11
 Uptake and storage of transition metal for physiological processes in our body is
happening through many complex formation reactions.Metallo proteins like blue copper proteins,
electron transfer proteins, iron-sulphur proteins are some examples. Iron-heme complex is
playing a vital role in oxygen transport. Recent studies reveals that around 40% of all known
enzymes require atleast one metal ion for its activity. Coordination compounds also play an
important role in serving humanity, because of their wide range of applications in health care
from antimicrobial to anticancer, bioengineering, biomimetic models, catalysis, photosensitized
materials etc.

When a solution of copper sulphate is mixed with an excess of ammonia solution, initially a
white precipitate of copper hydroxide is formed, and it is dissolved again to give a deep blue
solution of cuprammoniumsulphate complex. The complex is re-precipitated from the solution
by the addition of alcohol in which it is insoluble.

CuSO4 + 4NH4OH → [Cu (NH3)4] SO4. H2O (Tetrammine copper sulphate

monohydrate)
[Cu (NH3)4]2+
Tetrammine Copper (II) ion

12
Procedure

 In a 250 ml beaker, take 5 ml of saturated solution of copper sulphate.

 To this solution, add 1: 1 ammonia solution until a clear blue liquid is obtained.

 If there appears a white precipitate or turbidity, a little more ammonia solution is added
to dissolve it.

 Now, 15 ml of alcohol is added in drop wise with constant stirring till a purple
precipitate of cuprammoniumsulphate is obtained.

 The compound formed is filtered, dried, and weighed.

Calculation

Mass of CuSO4.5H2Otaken in 100 ml = 32 g

Mass of CuSO4.5H2Orequired for 5 ml =

Molecular mass of Copper Sulphate pentahydrate = 249.68 g/mol

No. of moles of Copper Sulphate = g of CuSO4.5H2O taken / Molecular weight (g/mol)

= moles

As Copper Sulphatepentahydrate is the limiting reagent, No. of moles of CuSO 4 = No. of moles
of Complex formed

13
Molecular mass of the Complex [Cu (NH3)4] SO4. H2O] = 245.74 g

Theoretical yield of the complex = No. of moles x Molecular weight (g/mol)

= g

Actual yield of the complex = g

Percent yield = Actual yield x 100

Theoretical yield

Result
Percent yield of TetrammineCopper (II) Sulphate Monohydrate= _________

                                                                                          

EXPERIMENT – 3

Estimation of Ferrous Sulphate by Permanganometry

Objective

To determine the amount of the given FeSO4.7H2O solution.

14
In this procedure 3 steps are involved:

i. Preparation of standard 0.05M FeSO4. 7H2O solution

ii. Standardization of KMnO4 solution using standard 0.05M FeSO4. 7H2O solution
iii. Titration of unknown FeSO4. 7H2O solution against standard KMnO4 solution.

Requirements
(a) Apparatus

Volumetric flask- 250ml, Pipette, -25ml, Watch glass, Funnel, Wash bottle, Burette, Conical
flask

(b)Chemicals

Standard FeSO4.7H2O (to be prepared)

FeSO4.7H2O solution of unknown concentration (Will be given)

Dilute H2SO4 solution

Potassium permanganate solution

Theory
Potassium permanganate (KMnO4) in acidic solution is a powerful oxidizing agent,
widely used in titrimetric analysis. In the presence of dilute sulphuric acid, KMnO 4 reacts with
the reducing agent ferrous sulphate. MnO4 - reacts with Fe2+ ions in an acidic solution to give
Mn2+ and Fe3+ as products. The equation for this redox reaction is:
MnO4 - (aq) + 8 H+ (aq) + 5 Fe2+ (aq)  Mn2+ (aq) + 5 Fe3+ (aq) + 4H2O

When all the FeSO4 has been oxidized, the excess of KMnO 4 added is not decomposed
and imparts pink color to the solution. This way it acts as a self-indicator.H2SO4 is most suitable
to provide an acidic medium because it has no action on KMnO4 in dilute solution. Other mineral
acids like Hydrochloric acid & Nitric acid are not suitable as they interfere in the redox reaction
between MnO4 -& Fe2+ ions. HCl can liberate chlorine gas by reacting with KMnO 4, which may
oxidize ferrous ions. Nitric acid itself is an oxidizing agent so it can replace KMnO 4 in the redox
reaction

15
 KMnO4 is not used as a primary standard because of the following reasons. It cannot be
obtained in a high state of purity as MnO2 is present in trace amounts as impurity. It readily reacts
with any traces of organic material or any other reducing substance in water. It decompose in the
presence of sunlight as follows:-

4MnO4─+ 2H2O  4MnO2+ 3O2 + 4OH ─

Permanganate is also unstable in the presence of manganese (II) ions:

4MnO4─ + Mn2+ + 2 H2O  5MnO2 + 4H+

But this reaction is very slow in acidic medium.

KMnO4 solution should be protected against unnecessary exposure to sunlight and so is always
kept in dark–colored bottles.

Hence before it can be used for any titrimetric analysis, KMnO 4 solution is first
standardized. For standardizing KMnO4 standard ferrous sulphate solution can be used. Ferrous
sulphate is easily prone to atmospheric oxidation, which can be prevented by dissolving the
crystals in dilute sulphuric acid.

Procedure

Preparation of Standard 0.05M FeSO4.7H2O solution.

1. Weigh out between 1.4 g. of FeSO4. 7H2O on a watch glass.

2. Transfer this weighed amount of FeSO 4. 7H2O completely into the 100 ml volumetric
flask, carefully by the means of a funnel. Add 10 ml (approx.) of dilute H 2SO4 to it. If any
crystals are sticking to the watch glass or the neck of the funnel or flask, then use the
minimum amount of the distilled water to transfer them completely into the flask.
3. Remove the funnel and dissolve the FeSO4. 7H2O completely.
4. Add enough distilled water into the standard flask and make up the volume to the mark
on the neck of the flask.
5. Calculate the Molarity of this solution.

16
(b) Standardization of KMnO4 solution using standard 0.05M FeSO4. 7H2O solution

1. Fill the burette (after rinsing) with the given potassium permanganate solution.
2. Rinse the pipette with FeSO 4. 7H2O standard solution. Pipette 25ml of this standard
solution in a conical flask.
3. Add 10 ml (approx.) of dilute H2SO4 to this.
4. Titrate, by adding the potassium permanganate from the burette to the conical flask, with
constant stirring, until a light pink coloration is obtained. This is the end point.
5. Repeat this titration till you get 2 concordant readings. Record your readings in the table
given below
6. Calculate the molarity of the potassium permanganate solution using the equation.
(c) Titration of Unknown FeSO4. 7H2O solution against Std. KMnO4 solution

1. Titrate the unknown FeSO4. 7H2Osolution, provided against the potassium permanganate
solution, in the same way as described above.

2 Repeat this titration for concordant readings. Record your readings in the table given.
3. Calculate the concentration of the unknown FeSO4. 7H2O solution.

Observations and calculations Date: _____________

(a) Standardization of KMnO4

S.No. Vol. of FeSO4 Burette reading

17
 (ml) Initial Final Vol. of KMnO4 (ml)

Weight of FeSO4.7H2Oused to prepare standard solution (w) in g =

Molarity of standard FeSO4.7H2O solution M2 = (w) g            x 1000

                                                                                           278 x 100

Volume of FeSO4V2=

Volume of KMnO4 V1=

Molarity of KMnO4SolutionM1=M2 V2
V1

(b)Estimation of Unknown FeSO4.7H2O

S.No. Vol. of FeSO4 Burette reading

(ml) Initial Final Vol. of KMnO4 (ml)

18
Volume of FeSO4V2 =

Volume of KMnO4 V1=

Molarity of Std. KMnO4 M1=

Molarity of unknown FeSO4M2= M1 V1

V2

Amount of FeSO4.7H2O present per liter = Molarity of FeSO4.7H2O x 278

Result

Amount of the given FeSO4.7H2O solution = ____________________g/l

EXPERIMENT – 4

Determination of the Hardness of water by EDTA

Objective

19
To determine the temporary, permanent, and total hardness of water by EDTA titration.

Requirements

(a) Apparatus
Pipette, 25ml, Pipette filler, conical flask, Burette, 50 ml, Stand with clamp

(b) Chemicals

EDTA solution, Water sample, Eriochrome Black T indicator,

pH 10 buffer.

Theory

All-natural waters contain dissolved cations and anions. Water dissolves many ions as it flows
through minerals. Although water hardness is defined as the quantity of cations with a +2 or +3
charge, calcium ion and magnesium ion are the most common of such ions in natural water. The
formation of solid calcium carbonate is an endothermic process. Thus, when water containing
both carbonate and calcium ions are heated, calcium carbonate can precipitate out onto the walls
of pipes, boilers, and household items such as tea pots. This can shorten the lifetime of some of
these items. In addition, an insoluble scum develops when hard water comes into contact with
soap. Both calcium and magnesium ions are responsible for this precipitate. This scum can be
very difficult to clean. However, there is some evidence that hard water has beneficial health
effects. Selenium, for example, may help prevent cancer. Soft water drinking supplies have been
associated with an increased heart attack risk. Hard water contains dissolved magnesium and
calcium ions. These make it more difficult for the water to form lather with soap. Thus hard
water is responsible for the poor performance of similar cleaning agents.

Temporary hardness also called as carbonate or removable hardness is caused by

dissolved calcium and magnesium hydrogen carbonate (which is removed by boiling).
Permanent (non-carbonate or irremovable) hardness is caused by dissolved sulfates and chlorides
of calcium and magnesium (can’t be removed by boiling but require treatment). Water can be
softened using washing soda, lime soda, ion-exchange resin or zeolite process.

20
 Description Hardness (mg/L) or PPM
Extremely Soft 0-50
Soft 50-90
Moderately Hard 91-130
Hard 131-170
Very Hard 171-250
Extremely Hard Above250

EDTA is a versatile chelating agent.The quantity of “hardness ions” can be determined

by titration. EDTA, a weak acid, a hexadentate ligand will be used as the titrant. In its ionized
form, it is able to form soluble complexes with calcium and magnesium cations. The indicator
added to the sample is Eriochrome Black T. Initially, the indicator will form a complex with the
cations and is red in color. As the EDTA is added dropwise to the sample, it replaces the
Eriochrome Black T and forms more stable complexes with calcium and magnesium. When the
indicator is released by the metal ions, it has a distinct blue color. Therefore, the endpoint of the
titration is marked by the color change form wine red to blue. pH 10 buffer (boric acid
/potassium chloride /sodium hydroxide) is used in the titration.

Ca-EBT + EDTA4----> EBT + [Ca-EDTA] 2-

21
 Hardness is expressed as parts per million (mg per liter) of equivalent CaCO 3, as it is the
most insoluble salt. It’s molecular and equivalent weights are 100 and 50 respectively, makes the
calculations convenient.

For example, if the titration required V ml 0.01 M EDTA for 25 ml (0.025 L) of the water
sample, the calculation would be:

1 ml of 0.01 M EDTA ≡ 1mg of Calcium carbonate

Total Hardness of Water = Vol. of EDTA consumed in ml x Concentration of EDTA

Vol. of hard water in litres x 0.01

Procedure
(a) Standardization of EDTA Solution

Standardize EDTA using the given standard solution of hard water

1. Fill the burette with EDTA solution and record the initial burette reading.
2. Pipette out 25ml of the given standard hard water solution into a conical flask
3. Add 5 ml of pH 10 buffer solution to the conical flask followed by 5 to 6 drops
Eriochrome Black T indicator.
4. Immediately begin to titrate the sample. Be careful to titrate slowly near the endpoint, as
the color will take about 5 seconds to develop. The endpoint is the color change from
wine red to blue.
5. Record the initial and final burette reading.
6. Repeat the titration for concordant titre value.
7. Calculate the molarity of EDTA.

(b) Determination of Total Hardness of water

8. Fill the burette with standardised EDTA solution and record the initial burette reading.
9. Pipette out 25ml of the water sample into a conical flask
10. Add 5 ml of pH 10 buffer solution to the conical flask followed by 5 to 6 drops
Eriochrome Black T indicator.

22
 11. Immediately begin to titrate the sample. Be careful to titrate slowly near the endpoint, as
the color will take about 5 seconds to develop. The endpoint color change is from wine
red to blue.
12. Record the initial and final burette reading.
13. Repeat the titration for concordant titer value.
14. Calculate the total hardness of the water.
(c) Determination of Permanent Hardness of water

15. Fill the burette with standardised EDTA solution and record the initial burette reading.
16. Pipette out 25ml of the heated water sample into a conical flask
17. Add 5 ml of pH 10 buffer solution to the conical flask followed by 5 to 6 drops
Eriochrome Black T indicator.
18. Immediately begin to titrate the sample. Be careful to titrate slowly near the endpoint, as
the color will take about 5 seconds to develop. The endpoint color change is from wine
red to blue.
19. Record the initial and final burette reading.
20. Repeat the titration for concordant titer value.
21. Calculate the hardness of the water.

(d) Determination of Temporary Hardness of water

Total Hardness = Permanent hardness + Temporary hardness

Observations Date: _____________

(a) Standardisation of EDTA solution

S.No. Volume of Burette reading

standard hard Volume of EDTA (ml)
Initial Final
water (ml)

23
 (b) Determination of Total Hardness of water

S.No. Volume of Burette reading

Hard water Volume of EDTA (ml)
Initial Final
sample (ml)

(b) Determination of permanent Hardness of water

S.No. Volume of Burette reading

heated water Volume of EDTA (ml)
Initial Final
sample (ml)

24
Calculations

Molarity of EDTAsolution, M1V1 = M2V2

Determination of Total Hardness of water

Volume of EDTA solution consumed (V1) =
Molarity of EDTA solution (M1) =
Volume of Hard Water (V2) =
Molarity of Hard Water (M2) = M1V1

V2

Total hardness of given water sample = M2 x 1000 x 100 ppm of CaCO3

Determination of Permanent Hardness of water

Volume of EDTA solution (V1) =
Molarity of EDTA solution (M1) =
Volume of Hard Water (V2) =
Molarity of Hard Water (M2) = M1V1
V2

25
Permanent Hardness of given water sample = M2 x 1000 x 100 ppm of CaCO3

Determination of Temporary Hardness of water

Temporary Hardness = Total hardness- Permanent Hardness

Result
Total Hardness of Water = ppm
Permanent Hardness of Water = ppm
Temporary hardness of Water = ppm

EXPERIMENT - 5

Identification of Organic Compounds

Objective
Illustrate organic qualitative analysis using carboxylic acids & carbohydrates.

Requirements

(a)Apparatus
Test tubes, Test tube holder, Test tube rack, Wash bottle, Spatula, Beaker 100 ml.
(b) Chemicals:

26
(1) Samples to be tested:
Oxalic acid, Fructose, Succinic acid, Glucose, Salicylic acid,
Starch, Sucrose

(2) Reagents:
Sodium hydrogen carbonate

Calcium chloride solution, CaCl2 Glacial acetic acid, CH3COOH

Neutral Ferric Chloride Solution Resorcinol

Sulphuric acid (conc.) Sodium hydroxide solution

10% alcoholic solution of - naphthol (Molisch’sReagent) Conc.HCl

Fehling’s Solutions. (A and B) Dilute iodine solution

Theory
Carboxylic Acid

Among the organic compounds that show appreciable acidity by far the most important are the
carboxylic acids. These compounds contain the carboxyl group, which determines the
characteristic chemical behavior of compound. The carboxylic group is made up of a carbonyl
group (C=O) and a hydroxyl group (-OH). However, it is the (-OH) group that undergoes the
loss of H+ or replacement by another group.

O
-
R C O H + H2O RCOO + H3 O +

Carboxylic acid group is a polarized group, this result in carboxylic acids dissolving well in
water provided that their carbon chains are fairly short. Carboxylic acid group can form
hydrogen bonds with water molecule.

27
 O
R C
O H

The C+ is therefore likely to be attacked by nucleophile while O- may be attacked by an
electrophile. The C+ tends to attract electrons from the C-OH bond. The oxygen of the –OH
group in turn attracts electron from the O-H bond, weakening this bond and allowing the loss of
hydrogen as an H+ ion, making it acidic.

Carbohydrates
Carbohydrates [General Formula:CH2O)n ] include a diverse range of substances composed of
simple sugars such as glucose, fructose etc and high molar mass polysaccharides, such as
starch ,glycogen (animal starch) & other biopolymers of simple sugars.

When photosynthesis occurs in a plant cell, the energy from sunlight is converted to chemical
energy in a carbohydrate. This carbohydrate may be transferred to some other part of the plant
for use as an energy source. It may be converted to water – insoluble carbohydrate. For storage
until it is needed for energy or it may be transformed to cell wall material and become part of the
structure of the plant. If plant is eaten by an animal, the carbohydrate is used for energy by the
animal.

The simplest carbohydrates are the monosaccharide, also called simple sugars. Because they
have 6 carbon atoms simple sugars are sometimes called hexose. Glucose (formula shown
below) is the most common simple sugar involved in cell processes. Other simple sugars with
same formula but somewhat different structures are fructose, mannose, and galactose. These
must be changed to glucose before it can be used in a cell.

28
Units of two monosaccharide make up several very important sugars known as disaccharides.
Two molecules of monosaccharide join together to form a disaccharide.

C6H12O6 + C6H12O6 C12H22O11 + H2O

A molecule of water is lost. Disaccharides include sucrose (cane sugar used as a sweetener),
lactose (milk sugar), and maltose (a product of the breakdown of starch).

Polysaccharides consist of many simple sugar units hooked together. One of the most important
polysaccharides is starch, which is produced by plants for food storage. Animals produce a
related material called glycogen. The chemical formula of starch is (C 6H10O5) n, where n may
represent a number as high as several hundreds. What this means is that the very large starch
molecule consists of many units of C6H10O5joined together. For example, if n is 100, there are 6
times 100 carbon atoms, 10 times 100 hydrogen atoms, and 5 times 100 oxygen atoms in the
molecule. Its chemical formula is C600H1000O500. The atoms in a starch molecule are actually
present as linked rings represented by the structure shown below. Starch occurs in many foods,
such as bread and cereals. It is readily digested by animals, including humans.

29
Carbohydrates or polysaccharides are polyhydroxy aldehydes and ketones; within this broad
category are simpler compounds called sugars (or saccharides). The name carbohydrates are
derived historically from the idea, hydrates of carbon, that is the general formula C n (H2O) n. For
example, glucose has the formula C6H12O6 sucrose. (Common table sugar) is consistent with
another formula Cn (H2O)11. Carbohydrates do not have uniformly recognized properties.

These compounds are usually water-soluble solids, which melt with decomposition. These
characteristics correctly point towards the presence of a large number of highly polar functional
groups in these molecules. The presence of the hydroxyl group in a carbohydrate can be detected
by the Molisch’s reaction. Fehling’s solution will undergo reaction with reducing sugars such as
2-hydroxy ketones and 2-hydroxy aldehydes.

Procedure

Reactions of Carboxylic Acid.

(a) Oxalic Acid
(i)Identification Test: -Dissolve a small quantity of the compound in distilled water & add
sodium bicarbonate solution to it. Immediate effervescence indicates the presence of acidic
group (-COOH group) in Oxalic acid.

HOOC-COOH + NaHCO3 →NaOOC-COONa + H2O + CO2↑

(Effervescence)

(ii) Confirmatory Test: - To a little of the aqueous solution of the compound, add acetic acid
and CaCl2 solution. A white precipitate of calcium oxalate will be formed.

(COOH)2 + CaCl2 →Ca (COO)2 + 2HCl

(b) Salicylic Acid

(i)Identification Test: -To a small quantity of the compound in distilled water, add sodium
bicarbonate solution. Immediate effervescence indicates the presence of acidic group (-COOH
group) in Salicylic acid

30
C6H4 (OH) (COOH) + NaHCO3 → C6H4 (OH) COONa +H2O + CO2↑
(Effervescence)

ii) Confirmatory Test: - To a little of the compound, add neutral ferric chloride solution. Violet
Color will be observed. Salicylic acid reacts with ferric chloride to form ferric salicylate and
hydrochloric acid.

(c)Succinic Acid
(i) Identification Test: -Dissolve a small quantity of the compound in distilled water & add
sodium bicarbonate solution to it. Immediate effervescence indicates the presence of acidic
group (-COOH group) in Succinic acid
HOOCCH2-CH2COOH + NaHCO3 → NaOOCCH2- CH2COONa +H2O + CO2↑
(Effervescence)

ii) Confirmatory Test: - In a dry test tube take small amount of the compound and 1-2 crystal of
resorcinol; add 2-3 drops of conc. H 2SO4. Heat gently till the mixture become reddish brown.
Add 1 drop of this into about 50 ml of tap water, taken in a small beaker & add 2-3ml of NaOH.
A green yellowish fluorescence can be observed.

HO O O NaO O O
OH OH
CH2 COOH conc.H2SO4 NaOH
2 + CH2 COOH Heat

COOH COONa

Reactions of Carbohydrates.

(1) Molisch’s Test (Identification of Carbohydrates): This is an identification test for all
carbohydrates indicating the presence of polyhydroxyl group of carbohydrates. All carbohydrates
will answer this test.To an aqueous solution of the compound add 2 drops of 10 % alcoholic
31
solution of - naphthol and 3-4drops of conc. H 2SO4, through the sides of the test tube. A red
ring changing to violet at the junction of liquid layers indicates carbohydrate.

(2) Fehling’s Test (Distinction of Reducing & Non reducing Sugar): Glucose & Fructose will
answer this test while sucrose & starch will not. To an aqueous solution of the solid add few
drops of fehling (fehling A + fehling B) solution and heat by keeping in a water bath for a
minute. A red precipitate indicates reducing sugar.
OHC (CHOH)4CH2OH + 2Cu (OH)2 + NaOH → NaOOC (CHOH)4CH2OH+ Cu2O + 3H2O

(3) Iodine Test (Distinction of Starch & Sucrose) To an aqueous solution of the solid add few
drops of Iodine solution. Blue colour indicates the presence of starch & no blue colour for
sucrose.
Iodine forms a blue coloured complex with the amylose, which is water soluble part of starch.
(4) Distinction between Glucose and Fructose . To a concentrated aqueous solution of the
substance add 3 drops of conc.HCl and a small amountof resorcinol and boil for 5 mints.
Appearance of red colour confirms fructose & no red colour for glucose.

Observations: (Write the corresponding equations for the test) Date:

32
(a)CARBOXYLIC ACIDS:

E O I
x b n
p s f
e e e
Salicylic Acid
r r r
Identification test:
i v e
m a n
e ti c
n o e
t n

O
x
a
li
c
A Confirmatory test:
c
i
d

I
d
e
n
ti
fi
c
a
33
ti
o
n
t
e
st

C
o
n
fi
r
m
a
t
o
r
y
t
e
st

34
35
Succinic acid

Identification test

Confirmatory test

36
Experiment Observation Inference
Identification of carbohydrate

Distinction of reducing and non-

reducing sugars.

37
Experiment Observation Inference
Distinction of Sucrose and Starch

Distinction of Glucose and Fructose

38
Result

39
 EXPERIMENT – 6

Determination of Ionization constant of a weak acid

Objective

To determine the ionization constant of Aspirin (weak acid) by potentiometric titration

Requirements
(a) Apparatus

Dry Beakers, 100ml Pipette, 25ml

Burette, 50 ml pH meter

Wash bottle Pipette filler.

Conical flasks Stand with clamp

(b) Chemicals

pH standard buffer solutions Distilled water, 95% Ethanol

Sodium Hydroxide (0.1M) Weak acid (Aspirin 0.1M or 0.36 g/100 mL)

Theory

The pH of a solution signifies the H3O+ ion concentration in moles per litre. The pH of a
solution slightly decreases with the rise in temperature. Once the temperature is increased the
degree of dissociation of water will also increase. This will make the concentration of hydronium
ions to raise.

Neutralization reactions can be followed by measuring the pH with a pH electrode. This

produces a voltage related to the [H+ (aq.)] in the solution. The pH values of solution can be
accurately displayed: on a pH meter, on a chart recorder, on a data logger, on a computer to
produce a graphical output directly.

40
Weak Acid - Strong Base Titration:

Acidity constants play an important role in acid-base titrations, for we can use them to
decide the value of the pH that signals the stoichiometric point, the stage at which a
stoichiometrically equivalent amount of acid has been added to a given amount of base. The plot
of pH of an analyte, against the volume of titrant added is called a pH curve.

The given graphs show the pH curve for the titration of a weak acid (such as aspirin) with
a strong base (sodium hydroxide). At the stoichiometric point the solution contains acetyl
salicylate ions, sodium ions and the other ions generated from autoprotolysis. Due to the
presence of Bronsted base in the solution, we can expect pH > 7. Since the acid is weak, HA is
more abundant than A- ions in the solution. So, it furnishes more H 3O + ions by autoprotolysis.
After stoichiometric point, the OH- ions dominate.

Henderson- Hasselbalch equation

For a weak acid, this dissociates as follows: HA H + + A-

log10Ka = log10 [H+] + log10 [A-] - log10 [HA]

Therefore             -log10 [H+] = -log10Ka + log10 [A-] - log10[HA]

     

i.e., pH = pKa - log [acid]__

[conjugate base]

Halfway to stoichiometric point, when enough base has been added to neutralize half the acid,
the concentrations of acid & conjugate base are equal.

41
Therefore, pH = pKa - log10(1) [log 10(1) =0]
                                pH = pKa - 0   

pH = pKa   

So, from this equation, ionization constant of acetic acid can be calculated, by knowing the pH of
halfway of stoichiometric point. The following graph shows the results obtained of acid-base
titrations.

42
 The first thing to notice about these curves is that the pH does not change in a regular
manner as the alkali is added. Each curve has almost horizontal sections where a lot of the alkali
can be added without changing the pH much. There is also a very steep portion of each curve
where a single drop of alkali changes the pH by several units. In a titration, the equivalence point
is the point at which there are exactly the same numbers of moles of hydrogen ions as there are
moles of hydroxide ions. In each of the titrations the equivalence point is reached after 25.0cm 3
of the alkali has been added. However, the pH at the equivalence point is not always seven. Note
that in each case, except that in weak acid/weak base titration, there is a large and rapid change
of pH at the equivalence point (i.e., the curve is almost vertical) even though this may not be
centered on pH 7. This is relevant to the choice of indicator for a particular titration.

Potentiometric Titrator

43
Procedure

1. Fill the burette with 0.1 molar sodium hydroxide solution.

2. Weigh 0.4 g of aspirin and transfer into a 250 ml beaker. Add 10 ml of 95% ethanol,
stir with a glass rod. When the solid has dissolved add 90 ml of de-ionized water. Stir
the mixture until it becomes homogeneous.

3. Place a pH probe in the solution and connect it to a pH meter.

Note: The pH probe should have been calibrated using suitable buffer solutions.

4. Add 1 ml portions of sodium hydroxide solution from the burette to the beaker,
stirring well between additions and recording the pH.

5. Near the endpoint the pH begins to rise rapidly. Take 5-6 readings further and stop.

Processing data
1. Plot a graph of pH against volume of 0.10 mol dm −3 sodium hydroxide solution
added.

2. From the graph, calculate the endpoint of the titration.

3. Check this against the expected value by calculating the number of moles of aspirin
used (relative molecular mass of aspirin = 180) and, therefore, the volume of 0.10
mol dm−3 sodium hydroxide solution needed to react with it in a 1:1 mole ratio.

4. From the graph, estimate the pH at the half-way point of the titration. This gives a
value for the pKa of aspirin.

5. Calculate the Ka of18.00

aspirin
16.00

14.00

12.00

10.00
pH/V

8.00

6.00

4.00

2.00

0.00
13.0 13.2 13.4 13.6 13.8 14.0 14.2 14.4 14.6 14.8 15.0

44
 Volume of Sodium Hydroxide

Tabulations & Calculations Date: ________________

Vol. of NaOH
S. No. pH ΔpH / / ΔV
(ml)
1

10

11

12

13

14

15

16

45
17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

46
Result

The ionization constant of Aspirin =

47
 EXPERIMENT - 7

Verification of Ostwald’s Dilution Law

Objective

To verify Ostwald’s dilution law and to determine the dissociation constant of acetic acid.

Requirements

(a) Apparatus

Conductivity meter Beakers

Burette Volumetric flasks

Pipette filler Wash bottle

(b) Chemicals

Acetic acid, 2M, 1M, M/2, M/4, M/8 and M/16

Theory

An electrolyte is a substance, which conduct electricity by the migration of ions. We can study
the migration of ions in solution by measuring the electrical resistance of a solution of known
concentration taken in a cell. To minimize the effect of electrolysis, alternating currentis used.
The resistance of a sample is proportional to its length and inversely proportional to its area of
cross-section.

48
 R  l/a

R =  l/a

Where  = specific resistance (resistivity), unit is ohm.cm, l/a is known as cell constant and unit
is cm -1
. The reciprocal of resistivity is called conductivity, expressed in the unit S cm -1
(1
Siemen= 1 ohm -1). Therefore, the conductivity of one cubic cm of solution is known as specific
conductivity, which is also the reciprocal of specific resistivity.

K = l / aR

An ion in solution is mobile and responds to the presence of an electric field. As an ion migrates
through a solution, it carries charge from one location to another and therefore gives rise to an
electric current. Our bodies are electric conductors and some of the thoughts you are currently
having as you read this sentence can be traced to the migration of ions through membranes in the
enormously complex electrical circuits of our brains.
The resistance, R (in ohms Ω), of the solution is related to the current, I (in amperes, A), that
flows when a potential difference, V (in volts, V), is applied between the two electrodes.

Ostwald’s Dilution Law

Consider the following equilibrium reaction for the dissociation of a weak electrolyte.
HA  H++A-
ka = [H+] [A-]

49
 [HA]Where ka represents the dissociation constant of the electrolyte. Consider C to
be the concentration of the electrolyte.If out of 1 mole, mole dissociates, the concentrations at
equilibrium of HA, H+& A – are C (1- ), C&
C respectively. Hence substituting these in the above equation, we get
ka = C2
1-
This equation representing the relation between Ka,& C is known as Ostwald’s dilution law.
can be calculated by the equation,  = c

c = 1000 K Where K is the specific conductance of the electrolyte,
C [ = 390.8 ohm -1 cm 2 mol-1 (for acetic acid)]
The degree of dissociation  tend to be unity, only at infinite dilution and at all other
concentrations it is less than 1. As the solution is diluted, the specific conductance (K) decreases
but molar conductance increases. On dilution, the dissociation of an electrolyte increases, and the
molar conductance increases, since the number of ions per unit volume decreases, specific
conductance decreases.
In case of weak electrolyte like acetic acid, molar conductance increases rapidly as concentration
decreases, and it cannot be found out by extrapolation. So, at infinite dilution molar conductance
will be the sum of ionic conductance at infinite dilution of the cation and anion. This is called
Kohlrausche’s law. The value of molar conductance at infinite dilution of acetic acid is 390.8
ohm-1 cm 2 mol-1 at 250C.

50
Procedure

 Take three fourth of 2M acetic acid in a 100ml beaker, which is rinsed with the same
solution and dip the conductivity cell into the solution.
 Measure the conductivity of the solution and record it.
 Rinse a 50 ml burette with this 2M acetic acid, measure out exactly 50 ml into a100ml
volumetric flask and make up to the mark; this is 1M acetic acid.
 Measure the conductivity of the above solution and record the readings in the table.
 Repeat step3 for making concentrationsM/2, M/4, M/8&M/16 (prepare each dilution in
100ml volumetric flask.)
 Measure the conductance of each of the above solutions and record the readings in the
table.
 Complete the calculations, and then plot a graph of m vs. C

51
 Calculations & Tabulations Date: _____________

Conc. of Specific c = 1000 К  = c Ka = C2 C

acid (C) Conductance(К) C  1-

2M

1M

M/2

M/4

M/8

M/16

Result

52
 (1) The dissociation constant of acetic acid =
(2) From the graph, it is found that Ostwald’s dilution law has been verified.

EXPERIMENT – 8

Determination of Solubility and Heat of Solution of Sparingly

Soluble Substance
Objective

1. To determine the solubility of sparingly soluble substance.

2. To calculate heat of dissolution for the process using Vant Hoff’s equation.

Requirements
(a) Apparatus

Burette, volumetric flasks – 100 ml & 25 ml, funnel, wash bottle, glass dropper, conical flask,
glass rod, beaker – 250 ml, spatula, filter paper, weighing balance.

(b) Chemicals

Sparingly soluble substance, oxalic acid, sodium hydroxide, phenolphthalein

Theory

The term solubility of a substance can be defined as the amount of substance dissolved in 100 g
of solvent to form a saturated solution at particular temperature.The solubility of solute depends
upon temperature and nature of solvent. Solubility of most of substances increases with
temperature.

A saturated solution is a chemical solution containing the maximum concentration of

a solute dissolved in the solvent. The additional solute will not dissolve in a saturated solution.
The amount of solute that can be dissolved in a solvent to form a saturated solution depends on a

53
variety of factors. The most important factors are temperature,pressure and chemical
composition.

A sparingly soluble substance is so-called because when it is stirred into water or mixed with
water, only a very small amount of the substance goes into the solution, and most of it remains
undissolved. The solution becomes saturated with that little amount of salt dissolved, and the salt

immediately dissociates into its ions. Examples for sparingly soluble substances are benzoic acid,
salicylic acid, adipic acid etc.

Solubility of organic acids can be determined by preparing saturated solution at different

temperatures followed by filtration. The filtrate can be titrated with a strong base sodium
hydroxide.

Solubility in g/L of solution

=(Normality of NaOH x Volume of NaOH xEq.wt.of organic acid )/ Volume of organic

acid
solution
Heat of Solution can be defined as enthalpy change associated with 1 mole of substance in large
excess of solvent. It can be evaluated by applying Vant Hoff’s equation as:

ΔH = [(RT1T2)/(T2-T1) ]ln (Solubility 2/ Solubility1)

Where ΔH = Change in enthalpy of the process

T1 andT2 = Absolute temperatures

R = Gas constant = 8.314 x 10-3kilojoule.K−1⋅Mol−1

Sodium hydroxide is not a primary standard. Therefore, it should be standardized with a suitable
primary standard such as oxalic acid. Sodium hydroxide reacts with oxalic acid according to the
following equation:

H2C2O4.2H2O +2 NaOH Na2C2O4 + 4H2O

54
Procedure

(a)Preparation of Standard 0.01Noxalic acid solutionsolution.

1. Weigh out around0.063 g. of oxalic acid on a watch glass.

2. Transfer this weighed amount completely into the 100 ml volumetric flask, carefully by
the means of a funnel. If any crystals are sticking to the watch glass or the neck of the funnel
or flask, then use the minimum amount of the distilled water to transfer them completely into
the flask.
3. Remove the funnel and dissolve the oxalic acid completely.
4. Add enough distilled water into the standard flask and make up the volume to the mark on
the neck of the flask.
5. Calculate the Normality of this solution.
(b) Standardization of NaOH solution using standard 0.01Noxalic acid solution

1. Fill the burette (after rinsing) with the given sodium hydroxide solution.
2. Rinse the pipette with oxalic acid standard solution. Pipette 25ml of this standard solution
in a conical flask.
3. Add 1 or 2 drops of phenolphthalein indicator to this.
4. Titrate, by adding the sodium hydroxide from the burette to the conical flask, with constant
stirring, until a light pink coloration is obtained. This is the end point.
5. Repeat this titration till you get 2 concordant readings. Record your readings in the table
given below.
6. Calculate the normality of sodium hydroxide solution using the equation.

55
 (c) Determination of Solubility

1. Set the water bath at a definite temperature.

2. Weigh 0.5 g of organic acid in a clean dry 250 ml beaker.
3. Add 100 ml of distilled water to it.
4. Keep it in the water bath for 2 minutes with constant stirring to prepare saturated
solution.
5. Remove the saturated solution from the water bath and filter the contents of it.
6. Transfer 10 ml of the filtrate to a conical flask.
7. Add 1 or 2 drops of phenolphthalein indicator to it.
8.Titrate, by adding the sodium hydroxide from the burette to the conical flask, with
constant
stirring, until a light pink coloration as end point is obtained.
9. Repeat the procedure for one or two more temperature.
10. Solubility can be calculated using the equation.

Observations and calculations Date: _____________

(a) Standardization of NaOH

S.No. Vol. of oxalic acid Burette reading
(ml)
Initial Final Vol. of NaOH(ml)

56
Weight of oxalic acid used to prepare standard solution (w) in g =

Normality of standard oxalic acid solution = (w) g            x 1000

                                                                                           63 x 100

Volume of oxalic acid =

Volume of NaOH=

Normality of NaOH Solution=

(b)Determination of Solubility

Solubility in g/L of solution

= (Normality of NaOH x Volume of NaOH x Eq.wt.of organic acid) / Volume of organic acid
solution

S.No. Temperature Volume of Sodium Solubility (g / L)

hydroxide (from burette)
(K)
(ml)

57
Show Solubility calculations:

(d) Calculation of Heat of Solution

ΔH = [(RT1T2)/(T2-T1) ]ln (Solubility 2/ Solubility1)

Result

1. Solubility of the given substance at different temperatures are:

a.

b.

2. Heat of solution, ΔH = kJ / mol

58
 EXPERIMENT No.9

Kinetics of Decoloration of Dye in Alkaline Condition

Aim:

1. To verify the order of the reaction with respect to dyes concentration.

2. To determine the rate constant of the reaction.

Requirements

(a)Apparatus and Instrument

Burette, pipette – 5 ml,pipette holder ,Amber bottle, small glass test tube, test tube stand,
Colorimeter

(b)Chemicals

Dye Solution, Sodium hydroxide solution

Theory:

A dye is a colored substance that chemically bonds to the substrate to which it is being
applied. This distinguishes dyes from pigments which do not chemically bind to the material
they color. Examples are Crystal Violet (hexamethylpararosaniline chloride), Fuchsin

59
 (rosaniline hydrochloride), phenolphthalein, Malachite Green, Rosaniline, etc. These are
organic compounds that contain a colored cation. The intense color of thecation is due to the
extended conjugated system of alternate single and double bonds. Uponreactionwith
NaOHthe conjugationgets disruptedandthecolor is lost. It can be explained in detail using
crystal violet as below:
ThereactionbetweencrystalvioletandNaOHisthefollowing:

Asimplifiedversionofthesamereactionis:

CV+ + OH–(aq)CVOH
(crystalviolet)(hydroxide)

Theratelawfor the above reactionisoftheform:

Rate=k[CV+]m[OH–]n,where
kis therateconstantforthereaction

mis theorderwithrespectto crystalviolet(CV+)

nis theorderwithrespecttothehydroxide ion.

If the [OH –]is more than 1000 times the concentration of the Dye (crystal violet).
[OH –] will be constant thus the modifiedformof theratelaw is,
Rate = (k [OH-]n) [CV+]m

Rate=k’ [CV+]m

60
 This constant k’ is sometimes referred to as the pseudo rate constant, because it does not
take into accounttheeffectof the NaOH.
Pseudorateconstant: k’=k[OH-] n

In thiswaythe[OH–]willnotchangeappreciablyduringthefiveminutes thattheexperimenttakes.
Since the absorbance of the cation follows Beer’s Law (that is to say, the absorbance is
directlyproportional to the cation’s concentration) the kinetics of the reaction can be
determined bymeasuringthe absorbance as afunctionof time.
A graph can be plotted between ln(Absorbance)andtime which will a give a linear plot
for the first order reaction.The rate constant, k’ can be determined from the slope of the
graph using the equation:
k’=–slope

Procedure
1. Switch on the Colorimeter and prepareablankbyfillinganemptycuvette or small

61
glass test tube 3/4fullwithdistilledwater.
2. Collect 3/4of cuvette or small glass test tube with the given dye solution.
3. Determine the absorbance for a range of lamda values by adjusting different
wavelengths in the colorimeter.
4. Plot the graph between wavelength and absorbance and locate λ max.
5. Set the λ max value in the colorimeter.
6. Transfer 4 ml dye solution and 1 ml NaOH solution to the cuvette /small glass
test tube by starting the timer.
7. Tilt the cuvette and place it in the colorimeter.
8. Note down the absorbance immediately.

9. Absorbance should be checked at definite time intervals (6 to 8 readings should be

Taken)

10. Calculate ln absorbance.

11. Plot the graph with ln absorbance against time and then determine the rate constant
from the slope.
12. Verify the order of the reaction from the shape of the graph.

Tabulations Date: _____________

Determination of λ max of the given Dye

Sl.No. Wavelength (nm) Absorbance
1 410

2 430

3 470

4 490

62
5 520

6 540

7 580

8 610

9 640

Kinetics of Decoloration of the given Dye

S.No Time Absorbance ln absorbance

. (Minutes)

Calculations:
From the graph:

Rate constant, k’ = - slope

63
Results & Conclusions:

(a)Rate constant for decoloration of dye =

(b) From the graph, it is found that the order of the reaction has been verified.

EXPERIMENT No.10

Colorimetric Estimation of Dye Concentration in Water Sample

Aim:

1. To estimate the unknown concentration of dye in water sample.

2. To determine the molar absorption coefficient,ϵ of the dye concentration.

Requirements

(a)Apparatus and Instrument

Burette, pipette – 5 ml,pipette holder ,Amber bottle, small glass test tube, test tube stand,
Colorimeter

(b)Chemicals

Dye Solution, Distilled water

Theory:

Colorimetry (using Colorimeter), a measurement of the wavelength is used extensively for

identification and determination of concentrations of substances that absorb light.
Two fundamental laws are applied: that of a French scientist, Pierre Bouguer, which is also
known as Lambert’s law, relates the amount of light absorbed and the distance it travels through
an absorbing medium; and Beer’s law relates light absorption and the concentration of the
absorbing substance.

BEER – LAMBERT LAW:

64
where I0 is the intensity of light absorbed and I is the intensity of light transmitted.

Epsilon = Molar Absorption Coefficient

The term “molar absorption coefficient (ϵmolar)” refers to the absorbance of light per unit path
length and per unit of concentration.
The basic equation to calculate molar absorptivity (absorption coefficient) for a given
wavelength:
ɛ=A/lxC

where

A is the absorbance,l is the length of the light path in cm and C is the concentration.

Absorbance should be determined for the varying dye concentration. Unknown dye
concentration can be estimated from the graph plotted between concentration and absorbance.

Procedure:
1. Switch on the Colorimeter and prepareablankbyfillinganemptycuvette or small

65
 glass test tube 3/4fullwithdistilledwater.
2.Set the λ max value (from the previous experiment) in the colorimeter.

3.The samples are prepared as follows:

Sample No. Volume of Dye Volume of water

(ml) (ml)

Sample 1 1 4

Sample 2 2 3

Sample 3 3 2

Sample 4 4 1

Sample 5 5 0

4.Note down the absorbance of each concentration. An unknown concentration of

dye is also given

5.Its concentration can be found out by plotting a straight-line graph of Absorbance Vs

Concentration of Dye.
6. Molar absorption coefficient for each dye concentration can be calculated using the
equation.

66
Tabulations and Calculations : Date: _____________

Length of the light path = 1 cm

Concentration of the given dye = mol/L

Sample Volume of the Concentration of the Absorbance ɛ=A/lxC

No. Sample prepared sample,
( Lmol-1cm-1)
(ml) C(mol/L) =
(Concentration of the
given dye x Volume of
the sample)/5 ml
1

Unknow - From the graph

n

Result

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Ɛ of the dye =
Amount ofDye in the given water sample =-------------mol / L

DEMONSTRATION - 1

Determination of sodium by flame photometry

Introduction
Sodium (Na) is the major extracellular cation, and it plays a role in body fluid
distribution. Concentration of sodium ions inside the plasma (extracellular) is 130-145 mmol/l.
Higher and lower concentrations are referred to as hypernatremia and hyponatremia,
respectively.

When a solution containing cations of sodium is sprayed into flame, the solvent
evaporates, and ions are converted into atomic state. In the heat of the flame (temperature about
1800ºC), small fraction of the atoms is excited. Relaxation of the excited atoms to the lower
energy level is accompanied by emission of light (photons) with characteristic wavelength (Na:
589 nm). Intensity of the emitted light depends on the concentration of particular atoms in flame.

Instruments, reagents and glassware

Flame photometer, Stock solutions of Na+, c = 100 mg/l (100 ppm). Volumetric flasks
100 ml, 6 numbers. Glass pipettes: 1, 2, 10 ml.

Preparation of standard solutions

Standard solutions are prepared by dilution of stock solutions. Use different glass pipettes
and numbered 100 ml volumetric flasks and prepare the solutions according to the following
table:

Flask number 1 2 3 4 5 6
Vol. of Na stock 1 2 5 7 8 X

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solution (ml)
Conc. of solution 1 2 5 7 8 X
obtained (ppm)

Procedure
Let the instrument warm up for 5 minutes. Aspirate distilled water to the instrument.
Select the filter for element Na. Adjust the reading to zero. Aspirate the most concentrated
standard solution (solution number 5) and note down the reading. Aspirate distilled water – the
instrument should read 0. Aspirate standard solutions no. 4, 3, 2 and 1 and then the test solution.
Record the results. Aspirate distilled water for at least 5 minutes to clean the system.

Graphical Evaluation
Draw calibration curves for sodium on a graph sheet. Use concentrations as abscissa and
instrument readouts as ordinate values. Find concentration of sodium ions in test solution from
calibration curve.

Result
The amount of sodium present in the given test solution = ppm

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