Experiment No.

− 1
AIM: To study various glassware/apparatus used in chemistry laboratory.
References:
1. Furniss B. S. et. al., "Textbook of Practical Organic Chemistry". Publications (India) Pvt.
Ltd. 2006, 5th ed. P. no. 66-69.
2. Singh H., Kapoor V.K. "Practical Pharmaceutical Chemistry". Vallabh Prakashan 1 st ed. P.
no. 11-12.
3. Joaquín,G.I. et al., "Experimental organic chemistry: laboratory manual". Academic
Press, 2015, P. no. 74-79, & 91-93.

Requirements: Test tube, test tube stand, test tube holder, beaker, conical flask, round bottom
flask, condenser, funnel, separating funnel, volumetric flask, spatula, reagent bottle, dropper,
pipette, measuring cylinder, burette.

Theory: Description of various glassware used in Pharmaceutical Chemistry Laboratory

1. TEST TUBE, TEST TUBE STAND, TEST TUBE HOLDER: Test tubes are
composed of glass or plastic tubing, open at top usually with a rounded U-shaped bottom.
Test tubes are available in multitude of length & width, typically from 10-20mm wide
and 50-200mm long. The top often featured with flared edge to avoid the spilling out the
content. The purpose of test tube is to hold a small quantity of a chemical entity. The test
tube is made up of a specialized glass material that can withstand the heating and cold
temperature conditions.
Test tube stand/rack can be made up of metal wires, fiberglass or polypropylene. It is
available in different forms (classic rack, an interlocking cube form, a stackable form, test
tube drying rack). It is required to keep the test tubes containing any chemical under
experiment, aligned and readily available during experiment.
Test tube holder is used to hold the test tube containing the chemical substances during
experiment. Generally such experiments involves heating of a substance inside the test
tube. Therefore, the test tube is picked with the help of a holder, which holds the test tube
near to its opening.
2. BEAKER: It is a simple container meant for mixing, stirring and heating the solutions in
a laboratory. Beakers are cylindrical-shaped, closed flat bottom, open at the top with a
 spout for easy pour of liquids. It is available in different sizes ranging from milliliters to
liters. They are often graduated with the indications of specified volumes that can be
accommodated.
3. FUNNELS: The funnels are long and short stem are made up of usually glass material
designed for simple filtration. The long stem funnel has a 60º angle (between the stem
and the cone) for liquid content, while the short/ wide stem types are for transferring the
powders. The Buchner funnel is made up of porcelain and has a perforated porcelain
plate to support a filter paper.
Separating Funnel is a pear-shaped flask with a stopcock at the bottom. It is used for
liquid-liquid extraction processes. The funnel allows the separation of immiscible liquids
based on their densities.

TEST BUCHNER’S
TEST TUBE BEAKER FUNNEL SEPARATING
TUBE STAND FUNNEL FUNNEL

4. FLASKS: These are the common type of glassware used for various chemistry operations.
Erlenmeyer flask is a cone-shaped flask with a narrow neck and a flat bottom. It used for
mixing, heating, and storing solutions. It is also used for titrations. The narrow neck helps
prevent spills and allows for easier swirling of contents.
Florence flask (round bottom flask) possess a round-bottom with a single, narrow neck.
It is primarily used for heating liquids evenly and for boiling. The round bottom allows
 for uniform heating, making it suitable for use on heating mantles or in water baths. It’s
also used for reactions that require heating under reflux.
Volumetric flask is a flat bottomed flask with a long neck marked with a single
graduation line. It has a precise volume capacity. It is used for the preparation of standard
solutions. The accurate volume measurement makes it essential for preparing solutions
with exact concentrations.
Buchner flask is similar to similar to an Erlenmeyer flask but with a sidearm for
filtering. The sidearm allows for the connection of a vacuum line. It is used in
conjunction with a Buchner funnel for vacuum filtration in order to separate solids from
liquids. It allows for efficient separation of solids from liquids under reduced pressure.

FLAT ERLENMEYER IODINE BUCHNER’S

BOTTOM FLASK FLASK FLASK
FLASK

5. CONDENSER: It is a type of laboratory glassware designed for the cooling and

condensation of vapors. It typically consists of the following components:

• Structure: It has a straight, vertical tube with an outer jacket surrounding the inner tube.
The inner tube carries the vapor, while the outer jacket allows coolant (such as water) to
flow around the inner tube.
• Material: Made of glass or, less commonly, plastic. Glass is preferred for its ability to
withstand temperature changes and for its chemical resistance.
• Cooling: The outer jacket is usually equipped with inlet and outlet ports for the coolant.
• Uses: In the distillation process, solvent recovery and reaction cooling in reflux reactions
 6. REAGENT BOTTLE is a bottle with a wide neck, usually made of glass or plastic,
often with a screw cap or stopper. It is provided for the storage of chemicals and reagents.

VOLUMETRIC
CONDENSER
REAGENT BOTTLE
SPATULA DROPPER
FLASK

7. SPATULA: Spatula is a flat, broad blade with a straight or slightly curved edge. The blade is
often tapered to facilitate precise handling. The handle is usually elongated for a comfortable
grip. It is commonly made from materials such as stainless steel, plastic, or ceramic. Stainless
steel spatulas are preferred for their durability and resistance to chemical reactions. Spatulas
are available in various sizes and shapes, including small spatulas for fine work and larger
ones for handling bulk materials. Spatula is used for transferring chemicals, scraping,
spreading etc.
 PIPETTE MEASURING CYLINDER
BURETTE

8. DROPPER: It is a simple yet essential tool in a chemistry lab. It is used for dispensing
and transferring small quantities of liquids with precision. The dropper typically consists
of a long, slender tube or pipette body. It may be made of glass or plastic. At one end of
the dropper, there is a bulb (often rubber or plastic) that can be squeezed to create suction
and draw liquid into the tube. The opposite end (tip) of the dropper is a narrow, pointed
tip through which the liquid is filled up and dispensed.
9. PIPETTE: Pipette is employed for accurate measuring and transferring the specific
volumes of liquids. It comes in various designs and materials as per different
applications. The main part of a pipette is a slender, cylindrical tube designed to hold and
dispense liquid. It typically has graduated markings to indicate different volumes. The
end of the pipette is often pointed or tapered to facilitate precise liquid handling. They are
widely used in experiments and procedures where accuracy is critical. It provides
accurate measurement and transfer of liquid volumes, essential for reproducible results in
experiments.
10. MEASURING CYLINDER: It is also known as graduated cylinder used for measuring
the volume of liquids with moderate precision. It is a long, cylindrical tube with a flat
base and a spout for pouring. It is marked with graduation lines to indicate different
volumes. It is made from glass or durable plastic. Glass cylinders offer higher accuracy
and chemical resistance, while plastic cylinders are often used for their durability and
cost-effectiveness.
11. BURETTE: It is used for precise dispensing and measuring of liquids, especially in
titrations. It comprises a long, graduated glass or plastic tube with a narrow bore and a
 stopcock at the bottom for controlled liquid flow. They are often made from glass or
durable plastic. Glass burettes offer high precision and are resistant to chemical reactions,
while plastic burettes are often used for their durability. Burette is ideal for dispensing
exact volumes of liquids where accuracy is crucial, such as in quantitative analysis and
preparation of solutions. Requires careful handling and regular cleaning to maintain
accuracy. The stopcock must be checked for leaks and smooth operation.
RESULT: The experiment described the commonly used different glassware in the
pharmaceutical chemistry lab, their types, shapes, sizes and utility.

EXPERIMENT NO: 2

AIM: To perform the limit test for chlorides in the given sample.

References
1. Indian Pharmacopoeia Commission. "Indian Pharmacopoeia" (IP 2022). Government of
India. Vol. II. P. no. 172.
2. Ahmed H. et al. "Pharmaceutical Importance and Significance of Limit Tests." Asian
Journal of Pharmaceutical Research, 7.1 (2017), P. no. 30-34.
3. Singh H. & Kapoor V.K. "Practical Pharmaceutical Chemistry". Vallabh Prakashan 1st ed.
P. no. 66-68.
4. Chatwal G.R. "Pharmaceutical Chemistry-Inorganic". Himalaya Publishing House Pvt. ltd.
Vol. I, 5th ed. P. no. 48-49.

APPARATUS REQUIRED: Nessler cylinder, glass rod, beaker and pipette.

CHEMICALS REQUIRED: Standard Sodium Chloride solution, dilute Nitric acid solution, 0.1
M Silver nitrate solution, distilled water, sample.

THEORY: In Chemistry, Limit means a value or amount that is likely to be present in a substance
and test means to examine or to investigate. Thus, limit test is to identify the impurities in the
substance and compare it with standard. In general, limit test is defined as quantitative or semi
quantitative test designed to identify and control small quantities of impurity which is likely to be
present in the substance.
Limit test is generally carried out to determine the inorganic impurities present in compound.
Limit test of chloride is based on the reaction of soluble chloride with silver nitrate in presence of
dilute nitric acid to form silver chloride which appears as solid particles (Opalescence) in the
solution.

PRINCIPLE: This test is designed for the control of chloride impurity in inorganic substances. It
depends upon the precipitation of chlorides upon reaction with silver nitrate in the presence of
nitric acid. The extent of precipitation depends upon the amount of silver chloride formed i.e. on
the amount of chloride ions present in the substance. The opalescence produced was compared
with a reference standard opalescence having a known amount of chloride impurity.
Chemical Reaction Involved

Chloride Silver
Nitrate
ions chloride
ions
Role of Nitric acid: Nitric acid is used to remove any carbonate ions that might be present,
which can produce a white precipitate of silver carbonate, giving a false positive result for
chloride ions.

When nitric acid reacts with bicarbonates (such as sodium bicarbonate, potassium carbonate), the
reaction produces carbon dioxide gas, water, and a salt. Here's a general representation of the
reaction:

NaHCO3 + HNO3 NaNO3 + CO2 + H2O

Nitric acid is added in the limit test of chloride to make solution acidic and helps silver chloride
precipitate to make solution turbid at the end of process.

PROCEDURE:

STEP-1

1. Take two Nessler’s Cylinders, wash them thoroughly and then label as STANDARD and
TEST.
 2. Preparation of Standard chloride solution: Accurately weigh 0.05845g of sodium
chloride (NaCl) and dissolve in 100 ml of distilled water. (0.05845% w/v of NaCl).
3. Preparation of silver nitrate solution: Prepare 0.1 M silver nitrate (AgNO3) solution.
4. Preparation of dil. Nitric acid (HNO3) solution (10%): A small amount of dist. water was
added in a 100ml volumetric flask. Followed by addition of 10ml of HNO 3. Finally made
the volume upto the mark (100 ml) with distilled water.

STEP-2: Preparation of test and standard:

S. no. Test sample Standard compound
1 Specific weight of compound was 1ml of 0.05845 % W/V solution of
dissolved in water or solution is prepared sodium chloride was taken in Nessler
as directed in the pharmacopoeia and cylinder
transferred in Nessler cylinder
2 Added 10 ml of dilute nitric acid. Added 10 ml of dilute nitric acid.
3 Diluted to 50 ml as marked on Nessler Dilute to 50 ml as marked on Nessler
cylinder. cylinder.
4 Added 1ml of AgNO3 solution. Added 1ml of AgNO3 solution.
5 Kept aside for 5-10 min. Kept aside for 5-10 min.
6 Observed the Opalescence/Turbidity. Observed the Opalescence/ Turbidity.
OBSERVATION: The opalescence produced in sample solution should not be greater than
standard solution.
• If opalescence produces in sample solution is less than the standard solution, the sample
will pass the limit test of chloride.
• If opalescence produces in sample solution is more than the standard solution, the sample
will fail the limit test of chloride.

RESULT: The given sample (NaHCO3) passed/failed the limit test for chloride ions.

VIVA QUESTIONS:

1. What is impurity?

2. Define limit test.

3. What type of apparatus is used for performing limit test?

4. Write the principle involved in the limit test for chlorides.

5. How is standard solution prepared?

6. What type of reagent used in limit test for chlorides?

7. On what criteria the standard and test solutions are compared?

8. What is the purpose of dilute nitric acid?

9. What forms the precipitate in limit test for chlorides?

10. Define opalescence.

11. What is turbidity?

EXPERIMENT NO: 3

AIM: To perform the limit test for sulphate in the given sample.

References
5. Indian Pharmacopoeia Commission. "Indian Pharmacopoeia" (IP 2022). Government of
India. Vol. II. P. no. 174.
6. Ahmed H. et al. "Pharmaceutical Importance and Significance of Limit Tests." Asian
Journal of Pharmaceutical Research, 7.1 (2017), P. no. 30-34.
7. Singh H. & Kapoor V.K. "Practical Pharmaceutical Chemistry". Vallabh Prakashan 1st ed.
P. no. 57-68.
8. Chatwal G.R. "Pharmaceutical Chemistry-Inorganic". Himalaya Publishing House Pvt. ltd.
Vol. I, 5th ed. P. no. 49-50.

APPARATUS REQUIRED: Nessler cylinder, glass rod, beaker and pipette.

CHEMICALS REQUIRED: Barium chloride (BaCl2) solution, potassium sulphate, dilute

hydrochloric acid (HCl) solution, distilled water, sample.

THEORY: In Chemistry, Limit means a value or amount that is likely to be present in a

substance and test means to examine or to investigate. Thus, limit test is to identify the impurities
in the substance and compare it with standard.

Limit test is defined as quantitative or semi quantitative test designed to identify and control
small quantities of impurity which is likely to be present in the substance.

Limit test is generally carried out to determine the inorganic impurities present in compound.
Limit test of chloride is based on the reaction of soluble sulphate with barium chloride in
presence of potassium sulphate to form silver chloride which appears as solid particles
(Opalescence) in the solution.

PRINCIPLE: This test is designed for the control of sulphate impurity in inorganic substances.
It depends upon the precipitation of the sulphate with barium chloride in the presence of HCl and
traces of barium sulphate. The turbidity obtained is compared with standard solution, which
contains the known amount of sulphate ions under the same experimental conditions.
Chemical Reaction Involved

Sulphate Barium
Chloride
ions sulphateions

Role of Hydrochloric acid: Hydrochloric acid helps to make solution acidic.

Role of Potassium sulphate: Potassium sulphate is used to increase the sensitivity of the test by
giving ionic concentration in the reagent.

Role of ethanol: Alcohol helps to prevent super saturation of barium sulphate.

PROCEDURE:
Preparation of BaSO4 reagent
1. Prepared a 2 % solution of BaCl2 (Solution I).
2. Prepared ethanolic sulphate standard solution by dissolving 0.10891% K 2SO4 in 3% ethanol
(Solution II).
3. Then mixed the two solutions, by taking 10 ml of Solution I and 15 ml of Solution II.
Preparation of test solution
1. Dissolve a specified quantity of substance in water or prepare solution as directed by I.P
and transfer to Nessler’s cylinder.
2. To this added 2 ml of dilute HCl and made the volume to 45 ml with water.
3. Then added 5 ml of BaSO4 reagent.
4. Stirred and kept it aside for 5 min.
Preparation of standard solution
1. Prepared a solution of 0.1089% w/v of K 2SO4 and 1ml was transferred in Nessler’s
cylinder
2. Then added 2 ml of HCl to the Nessler’s cylinder and made the volume to 45 ml with
H2O.
3. To this added 5 ml of BaSO4 reagent.
4. Stirred and kept aside for 5 min.
5. Finally compared the turbidity of two solutions
OBSERVATION: The opalescence produced in sample solution should not be greater than
standard solution.
• If opalescence produces in sample solution is less than the standard solution, the sample
will pass the limit test of sulphate.
• If opalescence produces in sample solution is more than the standard solution, the sample
will fail the limit test of sulphate.

RESULT: The given sample (Borax) passed/failed the limit test for sulphate ions.
VIVA QUESTIONS:
1. Write the principle involved in limit test for sulphates.
2. What forms precipitate in limit test for sulphates?
3. How barium sulphate reagent is prepared?
4. The standard solution is prepared with________.
5. What is the importance of dilute hydrochloric acid in limit test for sulphates?
6. What is seeding agent?
7. What is the importance of ethanol in limit test for sulphates?
8. Write reaction involved in limit test for sulphate EXPERIMENT NO: 4

AIM: To perform the modified limit test for chlorides in the given sample.

References
1. Indian Pharmacopoeia Commission. "Indian Pharmacopoeia" (IP 2022). Government of
India. Vol. II. P. no. 172.
2. Ahmed H. et al. "Pharmaceutical Importance and Significance of Limit Tests." Asian Journal
of Pharmaceutical Research, 7.1 (2017), P. no. 30-34.
3. Singh H. & Kapoor V.K. "Practical Pharmaceutical Chemistry". Vallabh Prakashan 1 st ed. P.
no. 56-57.
4. Chatwal G.R. "Pharmaceutical Chemistry-Inorganic". Himalaya Publishing House Pvt. ltd.
Vol. I, 5th ed. P. no. 48-49.

APPARATUS REQUIRED: Nessler cylinder, glass rod, beaker and pipette.

CHEMICALS REQUIRED: Standard Sodium Chloride solution, dilute Nitric acid solution, 0.1
M Silver nitrate solution, distilled water, sample (potassium permanganate).

THEORY: In Chemistry, Limit means a value or amount that is likely to be present in a substance
and test means to examine or to investigate. Thus, limit test is to identify the impurities in the
substance and compare it with standard. In general, limit test is defined as quantitative or semi
quantitative test designed to identify and control small quantities of impurity which is likely to be
present in the substance.

Limit test is generally carried out to determine the inorganic impurities present in compound.
Limit test of chloride is based on the reaction of soluble chloride with silver nitrate in presence of
dilute nitric acid to form silver chloride which appears as solid particles (Opalescence) in the
solution.

PRINCIPLE: This test is designed for the control of chloride impurity in inorganic substances. It
depends upon the precipitation of chlorides upon reaction with silver nitrate in the presence of
nitric acid. The extent of precipitation depends upon the amount of silver chloride formed i.e. on
the amount of chloride ions present in the substance. The opalescence produced was compared
with a reference standard opalescence having a known amount of chloride impurity.
Chemical Reaction Involved

Chloride SilverNitrate
ions chloride
ions
Role of Nitric acid: Nitric acid is used to remove any carbonate ions that might be present,
which can produce a white precipitate of silver carbonate, giving a false positive result for
chloride ions.

When nitric acid reacts with bicarbonates (such as sodium bicarbonate, potassium carbonate), the
reaction produces carbon dioxide gas, water, and a salt. Here's a general representation of the
reaction:

NaHCO3 + HNO3 NaNO3 + CO2 + H2O

Nitric acid is added in the limit test of chloride to make solution acidic and helps silver chloride
precipitate to make solution turbid at the end of process.

PROCEDURE:

STEP-1

1. Take 0.5mg of potassium permanganate (KMnO4), add 20ml of ethanol and heat until the
colour becomes colourless.
2. Filtered the solution and collected the filtrate to proceed for the limit test.

STEP-II

1. Take two Nessler’s Cylinders, wash them thoroughly and then label as STANDARD and
TEST.
2. Preparation of Standard chloride solution: Accurately weigh 0.05845g of sodium chloride
(NaCl) and dissolve in 100 ml of distilled water. (0.05845% w/v of NaCl).
3. Preparation of silver nitrate solution: Prepare 0.1 M silver nitrate (AgNO3) solution.
4. Preparation of dil. Nitric acid (HNO3) solution (10%): A small amount of distilled water
was added in a 100ml volumetric flask. Followed by addition of 10ml of HNO 3. Finally
made the volume upto the mark (100 ml) with distilled water.
STEP-3: Further procedure was as following table:
S.no. Test sample Standard compound
1 20 ml of sample solution was transferred 1ml of 0.05845 % W/V solution of sodium
to the Nessler cylinder chloride was taken in Nessler cylinder
2 Added 10 ml of dilute nitric acid. Added 10 ml of dilute nitric acid.
3 Diluted to 50 ml as marked on Nessler Dilute to 50 ml as marked on Nessler
cylinder. cylinder.
 4 Added 1ml of AgNO3 solution. Added 1ml of AgNO3 solution.
5 Kept aside for 5-10 min. Kept aside for 5-10 min.
6 Observed the Opalescence/Turbidity. Observed the Opalescence/ Turbidity.
OBSERVATION: The opalescence produced in sample solution should not be greater than
standard solution.
• If opalescence produces in sample solution is less than the standard solution, the sample
will pass the limit test of chloride.
• If opalescence produces in sample solution is more than the standard solution, the sample
will fail the limit test of chloride.

RESULT: The given sample (KMnO4) passed/failed the limit test for chloride ions.
Viva Questions
1. What is the use of potassium permanganate.
2. How the modified limit test for chloride is distinguishable from the generally used limit
test for chloride.

EXPERIMENT NO: 5

AIM: To perform the modified limit test for sulphate in the given sample.

References
1. Indian Pharmacopeia Commission. "Indian Pharmacopeia" (IP 2022). Government of
India. Vol. II. P. no. 174.
2. Ahmed H. et al. "Pharmaceutical Importance and Significance of Limit Tests." Asian
Journal of Pharmaceutical Research, 7.1 (2017), P. no. 30-34.
3. Singh H. & Kapoor V.K. "Practical Pharmaceutical Chemistry". Vallabh Prakashan 1 st
ed. P. no. 57-58.
4. Chatwal G.R. "Pharmaceutical Chemistry-Inorganic". Himalaya Publishing House
Pvt.
Ltd. Vol. I, 5th ed. P. no. 49-50.

APPARATUS REQUIRED: Nessler cylinder, glass rod, beaker and pipette.

CHEMICALS REQUIRED: Barium chloride (BaCl2) solution, potassium sulphate, dilute

hydrochloric acid (HCl) solution, distilled water, sample (potassium permanganate).
THEORY: Limit means a value or amount that is likely to be present in a substance and test
means to examine or to investigate. Thus, limit test is to identify the impurities in the substance
and compare it with standard. It is carried out to determine the inorganic impurities present in
compound.

Limit test is defined as quantitative or semi quantitative test designed to identify and control
small quantities of impurity which is likely to be present in the substance.

PRINCIPLE: This test is designed for the control of sulphate impurity in inorganic substances.
It depends upon the precipitation of the sulphate with barium chloride in the presence of HCl and
traces of barium sulphate. The turbidity obtained is compared with standard solution, which
contains the known amount of sulphate ions under the same experimental conditions.
Chemical Reaction Involved

Sulphate Barium
Chloride
ions sulphateions

Role of Hydrochloric acid: Hydrochloric acid helps to make solution acidic.

Role of Potassium sulphate: Potassium sulphate is used to increase the sensitivity of the test by
giving ionic concentration in the reagent.

Role of ethanol: Alcohol helps to prevent super saturation of barium sulphate.

PROCEDURE:

Preparation of BaSO4 reagent

1. Prepared a 1% solution of BaCl2 (Solution I).
2. Prepared ethanolic sulphate standard solution by dissolving 0.10891% K 2SO4 in 3%
ethanol (Solution II).
3. Then mixed the two solutions, by taking 10 ml of Solution I and 15 ml Solution II.
Preparation of sample solution
• 0.1 mg of sample was dissolved in 20 ml of ethanol with gentle heating until the colour of
the solution disappeared.
• Then the solution was filtered to collect the clear filtrate.

Preparation of test solution

1. 20 ml of sample solution was transferred to the Nessler's cylinder.
2. To this added 2 ml of dilute HCl and made the volume to 45 ml with water.
3. Then added 5 ml of BaSO4 reagent.
4. Stirred and kept it aside for 5 min.

Preparation of standard solution

1. Prepared a solution of 0.1089% w/v of K 2SO4 and 1ml was transferred in Nessler’s
cylinder.
2. Then added 2 ml of HCl to the Nessler’s cylinder and made the volume to 45 ml with
H2O.
3. To this added 5 ml of BaSO4 reagent.
4. Stirred and kept aside for 5 min.
5. Finally compared the turbidity of two solutions (test and standard).
OBSERVATION: The opalescence produced in sample solution should not be greater than
standard solution.
• If opalescence produces in sample solution is less than the standard solution, the sample
will pass the limit test for sulphate.
• If opalescence produces in sample solution is more than the standard solution, the sample
will fail the limit test for sulphate.

RESULT: The given sample (KMnO4) passed/failed the limit test for sulphate ions. Viva

Questions
1. How the modified limit test for sulphate is distinguishable from the generally used limit
test for sulphate.
 EXPERIMENT NO: 6

AIM: To perform the limit test for iron in the given sample.

References
1. Indian Pharmacopeia Commission. "Indian Pharmacopeia" (IP 2022). Government of
India. Vol. II. P. no. 173.
2. Ahmed H. et al. "Pharmaceutical Importance and Significance of Limit Tests." Asian
Journal of Pharmaceutical Research, 7.1 (2017), P. no. 30-34.
3. Singh H. & Kapoor V.K. "Practical Pharmaceutical Chemistry". Vallabh Prakashan 1st ed.
P. no. 58-59.
4. Chatwal G.R. "Pharmaceutical Chemistry-Inorganic". Himalaya Publishing House Pvt.
Ltd.
Vol. I, 5th ed. P. no. 51-52.

APPARATUS REQUIRED: Nessler's cylinder, glass rod, beaker, measuring cylinder and pipette.

CHEMICALS REQUIRED: Thioglycollic acid, citric acid, Ammonia solution, ferric ammonium
sulphate, sample (NaCl).

THEORY: Limit test is defined as quantitative or semi quantitative test designed to identify and
control small quantities of impurity which is likely to be present in the substance. Limit test is
generally carried out to determine the inorganic impurities present in compound.

The limit test of iron is based on the reaction between iron and thioglycollic acid in the presence
of citric acid in an ammonical solution. A deep reddish purple colour is formed. Ferrous
thioglycolate is colourless in acidic medium but in alkaline medium it gives purple colour.

PRINCIPLE: This test is based on the formation of purple colour by reaction of iron (Fe 2+) with
thioglycollic acid (HSCH2COOH) in a solution made alkaline with ammonia, and containing
citric acid. The intensity of colour produced is compared with a standard colour obtained by a
similar reaction in a solution containing a definite quantity of ferric ammonium sulphate. The
comparison of colours is done in Nessler's cylinders by observing transversely.
Chemical Reaction Involved

Role of Citric acid

• Prevents precipitation of iron with ammonia.
• Interference of other metal cations is eliminated by the 20% citric acid solution. It forms
a complex with other metal cations.
Role of thioglycollic acid: It increases the sensitivity of the limit test.

PROCEDURE:
S.no. Test sample Standard compound
1 1 g of sample was dissolved in 40 ml of 2 ml of standard solution of iron (ferric
water and transferred to the Nessler's ammonium sulphate) was taken in
cylinder. Nessler's cylinder and diluted with 40
ml of water.
2 To this added 2 ml of 20% w/v citric To this added 2 ml of 20% w/v citric
acid solution (iron free). acid solution (iron free).
3 Then added 0.1 ml of thioglycollic Then added 0.1 ml of thioglycollic acid
acid and mix. and mix.
4 The solution was made alkaline with The solution was made alkaline with
ammonia solution (iron free). ammonia solution (iron free).
5 Kept aside for 5-10 min. Kept aside for 5-10 min.
6 Observed the Opalescence/Turbidity. Observed the Opalescence/ Turbidity.

Observation: Finally the two solutions were compared by viewing transversely/ vertically.

RESULT: The given sample (NaCl) ……….. the limit test for iron.

VIVA QUESTIONS:
1. Write the principle involved in limit test for Iron.
2. Write reaction involved in limit test for Iron.
3. The standard solution is prepared with________.
4. What is the importance of ammonia in limit test for Iron?
5. What is the agent used to produce colour in limit test for Iron? 6. On what criteria the standard
and test solutions are compared?
7. What is the purpose of 20%w/v of citric acid?
9. What type of colour produced in limit test for Iron?