LABORATORY MANNUAL

B.PHARM, SEMESTER-IV
MEDICINAL CHEMISTRY-1
SUBJECT CODE: BP406P

NAME OF STUDENT :
ACADEMIC SESSION :
ROLL NO :
REGISTRATION NO :

AUROSRI INSTITUTE OF PHARMACEUTICAL

EDUCATION AND RESEARCH (AIPER)
At: Kadei, Po: Uchhapada, Ps: Tangi, Dist: Cuttack, Pin: 754022
www.aiperodisha.org
 CERTIFICATE

This is certified to be a bonafide record of the subject of Medicinal Chemistry-1 of subject

Code ………………………………. And the Experimental work done by
……………………………….………………………………. of Bachelor of Pharmacy
………………………………. Semester/year, bearing Roll no ………………………
& registration no ……………………………….. During the Academic Session ……………
At Aurosri Institute Of Pharmaceutical Education &Research.

Principal Subject Teacher

Submitted to BPUT / OSBP for t h e practical examination held in ……………….

month at Aurosri Institute of Pharmaceutical Education & Research, Kadei, Tangi,
Cuttack.

Date:

External Examiner Internal Examiner

 Contents
EXPT. DATE OF Page no. Signature of Marks
NO. EXPERIMENT EXPERIMENT NAME teacher obtained

1 Synthesis of substituted pyrazole. 1,2

2 Synthesis of Benzimidazole 3,4

3 Synthesis of Benzotriazole 5,6
4 Synthesis of Diphenyl Quinoxaline 7,8
5 Synthesis of Benzocaine 9,10
6 Synthesis of Phenytoin 11,12

7 Synthesis of phenothiazine 13,14

8 Synthesis of Barbituric Acid 15,16

9 Assay of Chlorpromazine 17,18

10 Assay of Phenobarbitone 19,20
11 Assay of Atropine 21,22
12 Assay of Ibuprofen 23,24
13 Assay of Aspirin 25,26
14 Assay of Furosemide 27,28
15 The partition coefficient of iodine in 29,30
chloroform and water
16 The partition coefficient of benzoic 31,32
acid in benzene and water
Experiment : Date:

SYNTHESIS OF SUBSTITUTED PYRAZOLE

Aim of the Experiment: To prepare 1,3-diphenyl pyrazole from diphenyl hydrazone and
vicinal diol.
Chemical Required: 1-benzylidene-2-phenyl hydrazine, ethylene glycol, Ferric chloride, Tert-
butyl hydroperoxide, Acetyl acetone, sodium chloride, Ethyl acetate, sodium sulphate
Apparatus Required: conical flask, beaker, measuring cylinder, etc.,
Principle: 1,3-substituted pyrazole is prepared by cyclization of diaryl hydrazone and vicinal
diol in the presence of ferric chloride and test-butyl hydroperoxide (TBHP) which is also called
regioselective synthesis of substituted pyrazole.
Reaction :

Procedure: About 4.55g of 1-benzyledene-2-phenyl hydrazine is dissolved in the solution of

25ml of vicinal diol and ferric chloride (5mol%). Then another solution of tert-butyl
hydroperoxide (5.3g) in 25ml of acetylacetone is added to it. The mix solution is maintained at
a temperature range of 90-100oC. The mixed solution is left to reach room temperature and
stirred for 6 hours. Then the content is poured into water and extracted with ethyl acetate 3
times. The combined organic solution is washed with water, then with a saturated solution of
sodium chloride, passed through sodium sulphate, and evaporated under a vacuum.
Uses: Can be used as an antibacterial and antiviral agent.
Calculation :
Here limiting reagent is o-phenyl diamine; hence yield should be calculated from the
amount taken.
C13H12N2 = Molecular formula of 1-benzyledene-2-phenylhydrazine

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C15H12N2 = Molecular formula of 1,3diphenylpyrazole

Molecular weight of 1-benzyledene-2-phenylhydrazine = 196 g/mole
Molecular weight of 1,3diphenylpyrazole = 220 g/mole
Theoretical yield:
196g of 1-benzyledene-2-phenylhydrazine forms 220 g 1,3diphenylpyrazole
Therefore, 4.55 g 1-benzyledene-2-phenylhydrazine will form …….? (X) g
1,3diphenylpyrazole
X =(196 × 4.5)/220 = 4.09 g
Theoretical yield = 4.09 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100

Report/ conclusion: The percentage yield of 1,3-diphenyl pyrazole is ___________% with

m.p. 1850C
References :
1. N.Panda ,A K Jena ,J Org.chem,2012,77,9401-9406

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SYNTHESIS OF BENZIMIDAZOLE
Aim of the Experiment: Prepare and submit Benzimidazole from o-Phenylenediamine
Chemical Required: Sodium Hydroxide, o-Phenylenediamine, Formic Acid.
Apparatus Required: Beaker, Round Bottom Flask, Measuring Cylinder, heating Mentle,
Hot Air Oven, Filter paper, Funnel
Principle: The principle involved in the synthesis of Benzimidazole is the Phillips
reaction involves the condensation of ortho phenylenediamines with organic acids in the
presence of dilute mineral acids to furnish Benzimidazoles.

Mechanism: Initially one of the amino groups is acylated with the organic acid in the
presence of mineral acid to furnish an N-acylated compound. In the next step, the other
nitrogen is also acylated by making a bond with the carbonyl carbon of the first acyl
group leading to ring closure.

Reaction

Procedure: Place 27g of o-phenylenediamine in 250 ml RBF and add 16 ml of 90%

formic acid. Heat the mixture in a water bath at 1000C for 2 hrs. Cool, add 10 % sodium

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Experiment : Date:

hydroxide solution slowly, with constant rotation of the flask, until the mixture is just
alkaline to litmus. Filter off the solid benzimidazole at the pump and wash with a little
cold water. Recrystallize from hot water and dry in a hot air oven or in the air.
Uses: Antifungal.
Calculation :
Here limiting reagent is o-phenyl diamine; hence yield should be calculated from the
amount taken.
C6H4(NH2)2 = Molecular formula of o-Phenylenediamine
C7H6N2 = Molecular formula of Benzimidazole
Molecular weight of o-phenyl diamine = 108 g/mole
Molecular weight of Benzimidazole = 118 g/mole
Theoretical yield:
108 g o-phenyl diamine forms 118 g Benzocaine
Therefore, 27 g o-phenyl diamine will form …….? (X) g Benzimidazole
X =(108 × 27)/118 = 24.71 g
Theoretical yield = 24.71 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100

Report/ conclusion: The Benzimidazole was prepared and submitted in the laboratory
and its percentage yield was found to be

Reference: - Page. No. 1162, Vogel’s Practical Organic Chemistry 6th Edition

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Experiment : Date:

SYNTHESIS OF BENZOTRIAZOLE

Aim of the Experiment:: To synthesize and submit benzotriazole from o-

phenylene diamine and report its percentage yield
Chemical Required: o-phenylenediamine, glacial acetic acid, sodium nitrite
Apparatus Required: Beaker, conical flask, measuring jar, water bath, ice bath, thermometer,
etc.,
Principle :

The sodium nitrite reacts with glacial acetic acid and liberates nitrous acid. The o-
phenylene diamine reacts with nitrous acid and produces diazonium ions. When
the structure and stereochemistry of diazonium ions are stable, intramolecular
nitrogen coupling occurs and forms benzotriazole directly.
Reaction :

Procedure :
Dissolve 10.8g of o-phenylenediamine in a mixture of 18 ml of glacial acetic acid
and 62 ml of water in a beaker. Stir until the solid dissolves, warm gently if
necessary. Cool the solution to 150C. Stir well and add a solution of 25g of sodium
nitrite in 25ml water. The reaction mixture becomes warm within 2-3 minutes
reaches a temperature of about 850C and then begins to cool. The color changes from
deep red to pale brown. Continue stirring for 15 minutes till the temperature falls

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Experiment : Date:

about 35-400C. Thoroughly chill in an ice bath for 30 minutes. Filter the product and
wash it with cold water.

Uses: Used in the bulk drug industry as an important intermediate compound.

It is the basic nucleus present in anthelmintic drugs like mebendazole, thiabendazole, etc.
Calculation: Here limiting reagent is o-phenylenediamine; hence yield should be
calculated from its amount taken.
C6H8N2 = Molecular formula of o-phenylenediamine
C6H5N3 = Molecular formula of benztriazole
Molecular weight of o-phenylenediamine = 108 g/mole
Molecular weight of benztriazole = 119 g/mole
Theoretical yield:
108 g o-phenylenediamine forms 119 g benztriazole
Therefore, 10.8 g o-phenylenediamine will form ………? (X) g benztriazole
X =( 119 × 10.8)/108 = 11.9 g
Theoretical yield = 11.9 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100

Report/ conclusion: Benzotriazole was prepared and submitted. The percentage yield was
found to be ---------

REFERENCE:

1. Practical medicinal chemistry by Dr. Devala Rao, page no:35.

2. Comprehensive practical organic chemistry by V.K.Ahluwalia
and Renu Aggarval, page no:121.

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Experiment : Date:

SYNTHESIS OF 2,3 DIPHENYL QUINOXALINE

Aim of the Experiment: To synthesize and submit 2,3 diphenyl quinoxalines from
To synthesize and submit 2,3-diphenyl quinoxaline from o-phenylenediamine and report its percentage
yield.

Chemical Required: o-phenylenediamine, benzil, rectified spirit

Apparatus Required: Beaker, conical flask, measuring cylinder, water bath
Principle :

Quinoxalines are a type of heterocyclic compounds. They are also known as benzopyrazines.

(Benzopyrazine) (Pyrazine)
Generally, quinoxaline is formed by the condensation of o-phenylenediamine with
diketones. Here 2,3-diphenyl quinoxaline is prepared by treating o-
phenylenediamine with benzil.
Reaction

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Procedure :
Add a solution of 1.1g of o-phenylenediamine in 8ml rectified spirit to a warm solution of
2.1g of benzil in 8ml rectified spirit. Warm the mixture for 30 minutes in a water bath. Add
water dropwise until slight cloudiness persists. Cool the solution and filter the product.
Uses :
Quinoxaline derivatives are used as antimicrobial agents like levomycin.
They are also used in dyes
Calculation :
Molecular weight of o-phenylene diamine =
The molecular weight of 2, 3-diphenyl quinoxaline=
------- g of o-phenylene diamine gives ------------------- g of 2, 3-diphenyl quinoxaline
1g of o-phenylene diamine =

Theoretical yield =

Practical yield =

Report/ conclusion:
The percentage yield of 2,3-diphenyl quinoxaline is found to be ______________

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SYNTHESIS OF BENZOCAINE

Aim of the Experiment: To prepare and submit Benzocaine from P-amino benzoic acid.

Chemical Required: PABA, Ethanol, Conc. H2SO4, dry HCl, Na2CO3 solution.
Apparatus Required: Reflux condenser, Two Naked Flask, Gas Inlet Tube, Beaker, Funnel,
Glass Rod, Vacuum Pump, Litmus paper
Principle :
Aromatic esters are prepared by esterification of aromatic acids with alcohol in the presence
of a Conc. H2SO4 or dry HCl which fastens the reaction. Benzocaine is an ester Which was
prepared by esterification of PABA with ethanol in the presence of HCl.
Reaction:

Procedure: Place 80ml of absolute ethanol in a 250-ml two-necked flask equipped with a
double surface reflux condenser and a gas inlet tube. Pass dry hydrogen chloride through the
alcohol until saturated. The increase in weight is about 20gm, remove the gas inlet tube,
introduce 12gm of p- p-aminobenzoic acid, and heat the mixture under reflux for 2 hours. Upon
cooling, the reaction mixture sets to a solid mass of the hydrochloride of ethyl p-
aminobenzoate. It is better, however, to pour the hot solution into 30 ml of water (no
hydrochloride separates) and add solid sodium carbonate carefully to the clear solution until it
is neutral to litmus. Filter off the precipitated ester at the pump and dry in the air.
Uses: Local anesthetic.

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Experiment : Date:

Calculation:
Here limiting reagent is p-amino benzoic acid; hence yield should be calculated from the
amount taken.
C7H7NO2 = Molecular formula of p-aminobenzoic Acid
C9H11NO2 = Molecular formula of Benzocaine
Molecular weight of p-aminobenzoic acid = 137 g/mole
Molecular weight of Benzocaine = 165 g/mole
Theoretical yield:
137 g p-aminobenzoic acid forms 165 g Benzocaine
Therefore, 12 g p-aminobenzoic acid will form …….? (X) g Benzocaine
X =(165 × 12)/137 = 14.45 g
Theoretical yield = 14.45 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100

Report/ conclusion: Benzocaine was prepared and submitted in the laboratory and its
percentage yield was found to be
Reference: - Page. No. 897, Vogel’s Practical Organic Chemistry, 6th Edition

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Experiment : Date:

SYNTHESIS OF PHENYTOIN

Aim of the Experiment: To prepare phenytoin from benzil and urea.

Chemical Required: Benzil, Sodium Hydroxide, urea, Ethanol, Conc.H2So4.
Apparatus Required: Round-bottom flask – 100 ml, Reflux condenser, Crystallizing dish –
500 ml, Heating mantle, Stirrer, Beaker – 400 ml, Filtering flask with Büchner funnel,
Graduated cylinders – 100 ml and 50 ml, Petri dish, etc.
Principle: A catalyzed reaction between benzil and urea is used for the synthesis of phenytoin.
The reaction proceeds via intramolecular cyclization to form an intermediate heterocyclic
pinacol, which on acidification yields hydantoin (phenytoin) as a result of the 1,2-diphenyl
shift in pinacol rearrangement reaction.
Reaction :

Procedure :
Place 5.3 g (0.025 mol) of benzil, 3.0 g (0.05 mol) of urea, 15 ml of aqueous sodium hydroxide
solution (30%), and 75 ml of ethanol in a round-bottomed flask of 100 ml capacity. Set up a
reflux condenser with the flask and boil using an electric heating mantle for at least 2 h. Cool
to room temperature, pour the reaction mixture into 125 ml of water and mix carefully.
Allow the reaction mixture to stand for 15 min and then filter the product under suction to
remove an insoluble by-product. Render the filtrate strongly acidic with concentrated
hydrochloric acid, cool in ice water, and immediately filter off the precipitated product under
suction. Recrystallize at least once from industrial spirit to obtain about 2.8 g (44%) of pure
5,5-diphenylhydantoin, m.p. 297-298 °C.
Uses: It is a common antiepileptic drug.
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Calculation :
Here limiting reagent is benzil; hence yield should be calculated from the amount taken.
C14H10O2 =Molecular formula of benzil
C15H12N2O2 =Molecular formula of phenytoin
Molecular weight of benzil = 210 g/mole
Molecular weight of phenytoin = 252 g/mole
Theoretical Yield:
210 g benzil forms 252 g phenytoin
Therefore, 5.3 g benzil will form …….? (X) g phenytoin
X =(252 ×5.3)/210 = 6.36 g
Theoretical yield = 6.36 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100
Report/ conclusion: Phenytoin was synthesized and the percentage yield was found to
be……….%.
REFERENCES
1. Vogel’s Textbook of Practical Organic Chemistry by Brian S. Furniss, Antony J.
Hannaford, Peter W. G. Smith & Austin R. Tatchell; Fifth Edition; Page No. 1153.
2. Practical in organic chemistry, by Hitesh G. Raval, Sunil L. Baldania and Dimal A.
Shah, Nirav Prakashan, Page No. 313.

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Experiment : Date:

SYNTHESIS OF PHENOTHIAZINE
Aim of the Experiment: To Prepare and submit Phenothiazine from diphenylamine.

Chemical Required: Diphenylamine, sulphur, anhydrous calcium chloride, alcohol

Apparatus Required: Beaker, conical flask, measuring jar, water bath, etc.,

Principle: Phenothiazine is prepared by fusing diphenylamine with Sulphur with the rapid
evolution of hydrogen sulphide.
Reaction :
H H
N N
S.CaCl2

S
(Diphenylamine ) (phenothiazine)
Procedure :
22 g of diphenylamine, 8.2 g of sulfur, and 3.2 gms. of anhydrous calcium chloride are
melted together. The reaction sets 140-150° C with the rapid evolution of hydrogen
sulfide; by lowering the temperature, a few degrees the reaction can be slackened. When
the reaction has moderated, the temperature is raised to 160° C for a time. The melt, when
cool, is ground up and extracted, first with water and then with dilute alcohol. The residue
consists of almost pure phenothiazine. It can be recrystallized from alcohol. Yield 93%,
yellowish leaflets; m.p. 180° C.
Uses: Antipsychotic
Calculation
Here limiting reagent is p-amino benzoic acid; hence yield should be calculated from the
amount taken.

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Experiment : Date:

(C6H5)2NH = Molecular formula of diphenylamine

C12H9NS = Molecular formula of Phenothiazine
Molecular weight of diphenylamine = 169 g/mole
Molecular weight of Phenothiazine = 199 g/mole
Theoretical yield:
169 g diphenylamine forms 199 g Phenothiazine
Therefore, 12 g diphenylamine will form …….? (X) g Phenothiazine
X =(169 × 22)/199 = 18.68 g
Theoretical yield = 18.68 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100

Report/ conclusion: Phenothiazine was synthesized and the percentage yield was found to be
_________ %
Reference: - Systematic organic chemistry, by W. M. Cumming, 325-326, 1937.

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SYNTHESIS OF BARBITURIC ACID

Aim of the Experiment: To prepare barbituric acid from urea and dimethyl malonate
Chemical Required: Sodium metal. Ethanol, Diethylmalonate, Urea, calcium Chloride,
Conc.HCl
Apparatus Required: Round bottom flask – 2000 ml, Reflux condenser, Beaker, Buchner
funnel, Measuring cylinder, Filter paper
Principle: The synthesis of barbituric acid is affected by condensation of diethyl malonate with
urea in the presence of sodium ethoxide which may be prepared by reacting Na metal with
ethanol and undergo cyclization reaction with diethyl malonate.
Reaction :
O
O

H2N OEt NH
EtO
O
+ H2N
O N O
O OEt H

(Diethyl malonate) (Urea) (barbituric acid)

Procedure: Assemble a double surface reflux condenser with a 2-litre round-bottomed flask,
and place 11.5 g (0.5 mol) of clean sodium. Mix 250 ml of absolute ethanol in a portion and if
the reaction is unduly vigorous, immerse the flask in ice. When all the sodium has completed
the reaction, add diethyl malonate 80 g (76 ml, 0.5 mol), followed by a solution of dry urea 30
g (0.5 mol) in 250 ml of hot (70 °C) absolute ethanol. Shake the mixture thoroughly, attach a
calcium chloride guard tube to the top of the condenser, start reflux of the mixture for 7 h in an
oil bath, and heat to 110 °C. A white solid will be separated. Treat the reaction mixture with
hot (50 °C) water 450 ml and then with concentrated hydrochloric acid, with constant stirring,
until the solution is acid (about 45 ml). Filter the resulting almost clear solution and leave it in
the refrigerator overnight. Filter the solid at the pump, wash it with 25 ml of cold water, drain
well, and then dry at 100 °C for 4 hours. The yield of barbituric acid is 50 g (78%). It melts
with decomposition at 245 °C.
Uses: Itself not active pharmacologically, but its derivatives are used as sleeping pills and
sedatives.

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Experiment : Date:

Calculation :
Here limiting reagent is diethyl malonate; hence yield should be calculated from the
amount taken.
The molecular formula of diethyl malonate = C7H12O4
Molecular formula of barbituric Acid = C4H4N2O3
Molecular weight of diethyl malonate = 160 g/mole
Molecular weight of barbituric acid = 128 g/mole
Theoretical yield:
160 g diethyl malonate forms 128 g barbituric acid
Therefore, 80 g diethyl malonate will form …….? (X) g barbituric acid
X =( 128 ×80)/160 = 64 g
Theoretical yield = 64 g
Practical yield = ————- g
% Yield = (Practical Yield)/(Theoretical Yield) × 100
Report/ conclusion: Barbituric acid was synthesized and the percentage yield was found to
be………...%
REFERENCES
1. Vogel’s Textbook of Practical Organic Chemistry by Brian S. Furniss, Antony J.
Hannaford, Peter W. G. Smith & Austin R. Tatchell; Fifth Edition; Page No. 1176.

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ASSSAY OF CHLORPROMAZINE

Aim of the Experiment: To carry out the Assay of Chlorpromazine

Chemical Required: Perchloric acid (0.1M), Chlorpromazine, mercuric acetate solution(5%
w/v in acetic acid), crystal violet solution (0.2%w/v in acetic acid), acetone, methyl orange,
conical flask, burette, beaker, potassium hydrogen phthalate, glacial acetic acid, crystal violet
indicator.
Apparatus Required: Pipette, Burette, Erlenmeyer Flask, Volumetric Flask (amber colored
& normal)
Principle: Chlorpromazine is estimated by non-aqueous titration which is suitable for titration
of weak acid and weak base. In this nonaqueous. Mercuric acetate is added in the non-aqueous
titration in order to remove the chloride ions. To prevent the interference of the chloride ion
released by the titrant. The mercuric acetate replaces the halide ion in chlorpromazine with
acetate ion which is a strong base. The endpoint is indicated by the appearance of a blue color.
Reaction :

Procedure :
1. STANDARDISATION OF PERCHLORIC ACID (0.1N)
Dissolved 0.5g of potassium hydrogen phthalate in 25ml of glacial acetic acid and added a few
drops of 5%w/v crystal violet indicator. Titrated the solution with 0.1N perchloric acid till blue
blue-green color appears.

2. ASSAY OF CHLORPROMAZINE

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Weighed accurately about 0.6g and dissolved in 200 ml of acetone. Added 15ml of mercuric
acetate solution. Titrated with 0.1M perchloric acid, using a saturated solution of methyl orange
in acetone as an indicator. Perform a blank determination and make a necessary correction.
Each ml of 0.1M perchloric acid is equivalent to 0.03553g of C 17H19ClN2S. HCl
Uses: schizophrenia and other psychoses like paranoia mania (overactive behavior), anxiety,
agitation, and dangerously impulsive behavior.
Calculation
a) Standardization of 0.1N perchloric acid solution Molarity of Perchloric Acid =
Weight (W)
Mol. wt of potassium hydrogen phthalate X Volume (V)
Where, W = Weight of potassium hydrogen phthalate (g)
V = Volume of NaOH solution consumed

a) Determination of Chlorpromazine
% purity of phenobarbitone = 0.03553 x V X Molarity (Calculated) x 100
Molarity (given) x W
Where Molarity (calculated) = Molarity obtained from step (a)
V = Volume of perchloric Acid used
0.03553 is the equivalent factor Molarity (given) = 0.1M
W = weight of sample

Report/ conclusion
The given sample contains …………………. mg of chlorpromazine.
References : Indian Pharmacopoeia 2018 Page no:1600-01.

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Experiment : Date:

ASSAY OF PHENOBARBITONE

Aim of the Experiment: To perform the Assay of Phenobarbitone

Chemical Required: Sodium Hydroxide, aldehyde-free ethanol, benzoic acid,
thymolphthalein solution, silver nitrate, pyridine and ether,
Apparatus Required: Pipette, Burette, Erlenmeyer Flask, Volumetric Flask
Principle: Phenobarbitone is assayed by non-aqueous titration. In this method, the drug is
dissolved in the pyridine and titrated with sodium hydroxide solution using thymolphthalein as
an indicator.
Reaction :

+ 2H2O

(Phenobarbitone ) (Sodium Hydroxide ) (Phenobarbitone Sodium)

Procedure :
1. STANDARDISATION OF SODIUM HYDROXIDE SOLUTION
Accurately weighed 0.6g of benzoic acid and dissolved it in a mixture of 30 ml of ethanol and
6 ml of water and titrated with ethanolic sodium hydroxide solution using 0.2 ml of
thymolphthalein as an indicator.
2. ASSAY OF PHENOBARBITONE
Weighed and powdered 20 tablets. Weighed a quantity of the powder containing about 0.1g
(100 mg ) of phenobarbitone in 5ml of pyridine added 0.25 ml of thymolphthalein solution and
10 ml of silver nitrate pyridine reagent and titrated with 0.1M ethanolic sodium hydroxide until
the purely blue color is obtained. Repeated the operation without the substance under
examination.

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The difference between the titrations represents the amount of sodium hydroxide required.
Equivalent factor: 1ml of 0.1M ethanolic sodium hydroxide=0.01161g of C12H12N2O3
Uses: antiepilepsy
Calculation
a) Standardization of 0.1M Sodium hydroxide solution
Molarity of NaOH= Weight (W)
Mol. wt of benzoic acid (122.12) X Volume (V)
Where W = Weight of benzoic acid (g)
V = Volume of NaOH solution consumed
b) Determination of Phenobarbitone
% purity of phenobarbitone = 0.01161 x V X Molarity (Calculated) x 100
Molarity (given) x W
Where Molarity (calculated) = Molarity obtained from step
(a) V = Volume of Sodium hydroxide used
0.01161 is the equivalent factor Molarity (given) = 0.1M
W = weight of the sample

Report/ conclusion: The percentage purity of Phenobarbitone was found to be =

Reference :
1. A textbook of Medicinal Chemistry-I, Pragi Arora, Varun Arora, Davinder Kumar,
Page no:282,283.
2. Indian Pharmacopoeia Volume III 2018,Page No:2899,2900.

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ASSAY OF ATROPINE

Aim of the Experiment: To perform the Assay of Atropine

Chemical Required: Erlenmeyer flask, Volumetric flask, Pipette, Burette etc. Hot Air Oven,
Apparatus Required: Perchloric acid (0.1M), atropine, glacial acetic acid, crystal violet
solution (0.2%w/v in acetic acid), acetone, methyl orange indicator, potassium hydrogen
phthalate
Principle: Atropine is assayed by non-aqueous titration which is generally used for the titration
of a weak acid with a weak base. In this titration non-aqueous solvent perchloric acid is used
and crystal violet is used as an indicator. At the endpoint blue color is obtained.
Reaction :

(Atropine ) (protonated atropine )

Procedure:
1. STANDARDISATION OF PERCHLORIC ACID (0.1N)
Dissolved 0.5g of potassium hydrogen phthalate in 25 ml of glacial acetic acid and a few drops
of 5%w/v crystal violet indicator. Titrated the solution with 0.1N perchloric acid till a blue-
green color appears.
2. ASSAY OF ATROPINE
Weighed accurately 400mg of atropine dissolved in 50ml of glacial acetic acid and added a
drop of crystal violet indicator. Titrated this solution with 0.1N perchloric acid until the green
color is obtained endpoint.
Uses: bradycardia, anesthetic

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Calculation

Report/ conclusion: The percentage purity of Atropine was found to be

References :
1. Indian Pharmacopoeia 2018 page no:1600-01.
2. A textbook of Medicinal Chemistry-I, Pragi Arora, Varun Arora, Davinder Kumar, Page
no:281,282

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ASSAY OF IBUPROFEN

Aim of the Experiment: To determine the percentage purity of Ibuprofen

Chemical Required: Ibuprofen, sodium hydroxide, Potassium hydrogen phthalate,
Phenolphthalein indicator, Phenol red, ethanol, etc.,
Apparatus Required: Erlenmeyer flask, Volumetric flask, Pipette, Burette etc.,
Principle: The principle involved in the assay of Ibuprofen is acid-base titration where the
acidic group in Ibuprofen is neutralized by titrating with base i.e. NaOH using phenolphthalein
as an indicator where the endpoint is colorless to pink.
Reaction:

Procedure :
1. STANDARDIZATION OF 0.1 M NaOH:
Weigh about 0.5gm of KHP into 250-mL Erlenmeyer flask which was previously powdered
and dried at 1100C. Dissolve the sample in about 30 mL of distilled water before you titrate.
Add five drops of phenolphthalein indicator and titrate with 0.1M NaOH by constant swirling
to the first appearance of a permanent pink color. Each mL of 0.1M NaOH is equivalent to
0.02042gm of C8H5KO4.
2. ASSAY:
Weigh accurately about 0.5gm of drug dissolved in 100ml of ethanol (95%) and titrate with a
0.1m NaOH using phenolphthalein as an indicator where the endpoint is a permanent pink
color. Repeat the titration with blank. Each mL of 0.1M NaOH is equivalent to 0.02663gm of
C13H18O2.

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Uses: Anti-inflammatory, painkiller

Calculation :

Report/ conclusion: The percentage purity of Ibuprofen was found to be

References : Indian Pharmacopoeia 2018 page no:2261-65.

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ASSAY OF ASPIRIN

Aim of the Experiment: To determine the percentage purity of Aspirin.

Chemical Required : Aspirin, sodium hydroxide, Potassium hydrogen phthalate(PHP/KHP),
Phenolphthalein indicator, Phenol red etc.,
Apparatus Required : Erlenmeyer flask, Volumetric flask, Pipette, Burette etc.,
Principle: The principle involved in the assay of Aspirin is acid base titration where the acidic
group in aspirin is neutralized by titrating with base i.e. NaOH and the excess base is back
titrated with an acid (HCl) using phenol red as an indicator where the endpoint is pink to
colorless.
Reaction :

Procedure
1. STANDARDIZATION OF 0.5 M NaOH: -
Weigh about 2.5gm of KHP into 250-mL Erlenmeyer flask which was previously powdered
and dried at 1100C. Dissolve the sample in about 30 mL of distilled water before you titrate.
Add five drops of phenolphthalein indicator and titrate with 0.5M NaOH by constant swirling
to the first appearance of a permanent pink color.
Each mL of 0.5M NaOH is equivalent to 0.1021gm of C8H5KO4.
2. STANDARDIZATION OF 0.5M HCL:
Pipette out 20 mL of 0.5m NaOH solution into 250mL Erlenmeyer flask add five drops of
phenolphthalein indicator and titrate with 0.5M HCl by constant swirling to the disappearance
of pink color.

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Experiment : Date:

3. ASSAY :
Weigh accurately about 0.5 gm of sample dissolved in 15m: of ethanol (95%), and add 50mL
of 0.5M NaOH. Boil gently for 10 minutes, cool, and titrate the excess alkali with 0.5M HCl
using Phenol red as an indicator. Repeat the titration with blank.
Each mL of 0.5M HCl is equivalent to 0.04504gm of C9H7O4.
Uses: heart attack, blood clot, angina
Calculation
Report/ conclusion

References :
1. A textbook of Medicinal Chemistry-I, Pragi Arora, Varun Arora, Davinder Kumar, Page
no:282,283.
2. Indian Pharmacopoeia Volume I 1996, Page No:69,70.
3. Pharmaceutical titrimetric analysis theory and practical, A.A Napoleon, Page No:11-
17.

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ASSAY OF FUROSEMIDE

Aim of the Experiment: To carry out the Assay of furosemide tablets.

Chemical Required: Furosemide, dimethyl formamide, sodium hydroxide, bromothymol
blue indicator,0.1N oxalic acid, Phenolphthalein indicator, potassium hydrogen phthalate
Apparatus Required: Erlenmeyer flask, Volumetric flask, Pipette, Burette, hot air oven ., etc
Principle: It is assayed by aqueous acid-base titration between weak acid furosemide and
strong alkali sodium hydroxide. In this assay, protophilic solvent dimethyl formamide is used
which enhances the acidity of furosemide so that it can be titrated with sodium hydroxide. To
make the effect of acid impurities present negligible a solvent blank determination is carried
out.
Reaction :

Procedure :
1. REPARATION AND STANDARDIZATION SODIUM HYDROXIDE:
40 g of Sodium hydroxide in sufficient water to produce 1000ml.
2. STANDARDIZATION OF 0.1M SODIUM HYDROXIDE SOLUTION:
Weighed accurately about 5g of potassium hydrogen phthalate previously dried at 120 0C for
two hours dissolved in 75ml of carbon dioxide-free water. Added 0.1ml of phenolphthalein
solution and titrate with the sodium hydroxide until a permanent pink color is produced.
Each ml of 0.1M NaOH is equivalent to 0.02042g of potassium hydrogen phthalate.
3. ASSAY METHOD BY (NEUTRALIZATION TITRATION) :
Weighed and powdered 20 tablets and weighed accurately about a quantity of powder
equivalent to 0.5g and dissolved in 40ml of dimethyl formamide and titrated with 0.1M sodium

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Experiment : Date:

hydroxide using bromothymol blue as an indicator the endpoint shows the color change from
yellow to blue. Carry out a blank titration.
4. ASSAY METHOD BY (UV SPECTROPHOTOMETRY):
Weighed and powdered 20 tablets and weighed accurately about a quantity of powder
equivalent to 0.1g of furosemide and shaken with 150ml of 0.1M sodium hydroxide for 10
minutes. Added sufficient 0.1M sodium hydroxide to produce 250ml and filter. Dilute 5ml to 200ml
with 0.1M sodium hydroxide and measure the absorbance of the resulting solution at the maximum at
about 271nm. Calculate the content of C12H11 ClN2O5S taking 580 as the value of A (1%, 1cm) at the
maximum at about 271 nm.

Uses: heart failure, liver or kidney disease, high blood pressure

Calculation
Report/ conclusion

References :

1. A textbook of Medicinal Chemistry-I, Pragi Arora, Varun Arora, Davinder Kumar,

Page no:288,289.
2. Indian Pharmacopoeia Volume III 2018,Page No:2899,2900.

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Experiment : Date:

The Partition Coefficient of Iodine In Chloroform And Water

Aim of the Experiment: To determine the partition coefficient of benzoic acid in benzene and
water.
Chemicals Required: chloroform, Iodine, Potassium Iodide, Sodium Thiosulphate
Apparatus Required: Beaker, Measuring cylinder, Volumetric flask, Separating funnel
Principle: When an excess amount of solid or liquid is added to a mixture of two immiscible
liquids while this substance is slightly soluble in both immiscible liquids. It will distribute itself
between the two phases until saturation if mixed by shaking vigorously. If an insufficient
amount of substance is added in two immiscible liquids to the saturation, it gets distributed in
two layers at a definite ratio, this phenomenon is called distribution law or partition law. The
ratio constant is called as partition coefficient or distribution coefficient. It is independent of
the total amount of the substance dissolved.
K = C1/ C2
Where K is a constant known as the distribution or partition coefficient.
C1 and C2 are the concentration of a solute in the two immiscible liquids.
Procedure: A solution of Chloroform in water was prepared. Then, 30 ml, 40 ml, and 50 ml of
this solution were transferred in stoppered flasks. To it, 20 ml and 10 ml of chloroform were
added to make a total of 50 ml. Then, 50 ml of water was added with vigorous shaking. The
two layers were then separated. 10 ml of chloroform layer was pipetted out and to it, 20 ml of
water was added and titrated with 0.1N sodium Thiosulphate and 0.02NN sodium Thiosulphate
solution using potassium Iodide as indicator. The procedure was then repeated with the aqueous
layer. The normality and concentration of benzoic acid were calculated.
Observation :
Volume of 0.1 sodium thiosulphate consumed
chloroform layer Aqueous layer
Flask 1
Flask 2
Flask 3

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Determination of K values:
Sl. No C chloroform C aqueous K = C chloroform /C aqueous
Flask 1
Flask 2
Flask 3

Result:

Conclusion:

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Experiment : Date:

The Partition Coefficient of Benzoic Acid In Benzene And Water

Aim of the Experiment: To determine the partition coefficient of benzoic acid in benzene and
water.
Chemicals Required: Benzoic acid, Benzene, Phenolphthalein, Sodium hydroxide
Apparatus Required: Beaker, Measuring cylinder, Volumetric flask, Separating funnel
Principle: When an excess amount of solid or liquid is added to a mixture of two immiscible
liquids while this substance is slightly soluble in both immiscible liquids. It will distribute itself
between the two phases until saturation if mixed by shaking vigorously. If an insufficient
amount of substance is added in two immiscible liquids to the saturation, it gets distributed in
two layers at a definite ratio, this phenomenon is called distribution law or partition law. The
ratio constant is called as partition coefficient or distribution coefficient. It is independent of
the total amount of the substance dissolved.
K = C1/ C2
Where K is a constant known as the distribution or partition coefficient.
C1 and C2 are the concentration of a solute in the two immiscible liquids.
Procedure: A solution of benzoic acid in benzene was prepared. Then, 30 ml, 40 ml, and 50
ml of this solution were transferred in stoppered flasks. To it, 20 ml and 10 ml of benzene were
added to make a total of 50 ml. Then, 50 ml of water was added with vigorous shaking. The
two layers were then separated. 10 ml of benzene layer was pipetted out and to it, 20 ml of
water was added, and titrated with 0.05 N sodium hydroxide solution using phenolphthalein as
indicator. The procedure was then repeated with the aqueous layer. The normality and
concentration of benzoic acid were calculated.
Observation :
The volume of 0.05 N NaOH consumed
Benzene layer Aqueous layer
Flask 1
Flask 2
Flask 3

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Determination of K values:
Sl. No C benzene C aqueous K = C benzene /C aqueous
Flask 1
Flask 2
Flask 3

Result:

Conclusion:

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THANK YOU

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