Experiment No:________ Date :

INDEX

Sr. Aim Page

No. No

1 To carry out synthesis of barbituric acid from diethylmalonate and urea. 2

2 To synthesize benzimidazole from o-phenylenediamine. 5

3 To synthesize benztriazole from o-phenylene diamine. 9

4 To Synthesis 5, 5 –diphenyl hydantoin (Phenytion) from Benzil and urea 13

5 To carry out synthesis of benzocaine from p-aminobenzoic acid. 18

6 To carry out synthesis of 2,3-diphenylquinoxaline from o-phenylene diamine. 21

7 To Synthesis phenothiazine from diphenylamine. 24

8 To Synthesis 1,3-diphenylpyrazole from diphenyl hydrazone and ethylene glycol. 27

9 To perform assay of aspirin as per IP2014. 31

10 To perform assay of Chlorpromazine as per IP2014. 37

11 To perform assay of phenobarbitone sodium as per IP 2014. 43

12 To perform assay of atropine. 48

13 To perform assay of ibuprofen as per IP2014. 52

14 To perform assay of ibuprofen as per IP2014. 56

15 To determine the partition coefficient for distribution of benzoic acid between 60

octanol and water.

16 To determine the partition coefficient for distribution of Salicylic acid between 63

octanol and water.

KBIPER Medicinal Chemistry-I Page 1

Experiment No:________ Date :

Experiment No: 1

Aim: To carry out synthesis of barbituric acid from diethylmalonate and urea.
Reference: Furniss BS., Hannaford AJ., Smith PW., and Tatchell AR. In Vogel’s Text Book of
Practical Organic Chemistry; 5th Edn; Pearson Education Private Limited, New Delhi, 2005, pp
1176.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder
Reagents and chemicals: diethylmalonate, urea, sodium metal, absolute ethanol, CaCl2
Principle: The synthesis of barbituric acid is effected by condensation of diethyl malonate with
urea in presence of sodium ethoxide which may be prepared by reacting Na metal with ethanol
and it undergo cyclization reaction with diethyl malonate.
Reaction:

Procedure:
 Place 1.3g sodium in 250 ml RBF and add 30 ml ethanol to dissolve it.
 Add 9.5 ml diethylmalonate. To this add 5.0 g urea in boiling ethanol.
 Reflux the reaction mixture for 1.5 hour on a boiling water bath. The solid ppts will be separated
out.
 Add 30 ml warm water. Acidify the reaction mixture with conc. Hydrochloric acid. Crude
precipitates of barbituric acid will be separated out. Filter the precipitates.
 Dry the product and weigh it. Report Theoretical yield, Practical yield, %Practical yield
and Melting point of crude and standard. Recrystallize from hot water, m.p. 245 °C

Calculation:
M.W. of diethylmalonate = 160 gm/mol
M.W. of barbituric acid = 128 gm/mol

 Theoretical yield:
M.W. of diethyl malonate = M.W. of barbituric acid
128 gm of diethylmalonate = 180 gm of barbituric acid
9.5 gm of diethylmalonate = (?)
= 180 X 9.5/ 128
= 13.35 gm of barbituric acid

KBIPER Medicinal Chemistry-I Page 2

Experiment No:________ Date :

 Practical yield = _____________gm (x gm)

 % yield
= Practical yield X 100 / Theoretical yield
= _____________ X 100 / 13.35
= ___________% w/w (y % w/w)
Result:

Sr
No.

1. Theoretical Yield of 13.35 gm

barbituric acid

2. Practical Yield of x gm
barbituric acid

3. % Practical Yield of y%
barbituric acid

4 Melting Point(oC) of
barbituric acid

5. Structure of barbituric
acid

KBIPER Medicinal Chemistry-I Page 3

Experiment No:________ Date :

 Study Question:
1 Give the structure of barbituric acid.

2 Barbituric acid comes under which class.

3 Give the use of barbituric acid.

4 Give the molecular formula of barbituric acid.

5 Give the molecular weight of barbituric acid.

6 What is Sedatives.

7 What is Hypnotics

8 Write the Principle of barbituric acid.

KBIPER Medicinal Chemistry-I Page 4

Experiment No:________ Date :

Experiment No: 2
Aim: To synthesize benzimidazole from o-phenylenediamine.

Reference: Furniss BS., Hannaford AJ., Smith PW., and Tatchell AR. In Vogel’s Text Book of
Practical Organic Chemistry; 5th Edn; Pearson Education Private Limited, New Delhi, 2005, pp
1162.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder

Reagents and Chemicals:

Sr. Chemical used Quantity required Total quantity required per

No. Batch of 30 students

1. o-phenylene diamine 6.7 gm 180 gm

2. Formic acid(90%) 4.7 gm(4 ml) 120 ml

3. Sodium hydroxide(10%) Add untill the mixture just q.s

alkaline to litmus

Procedure

 Place 6.7 gm of o-phenylene diamine in a 250 ml of conical flask, add 4.7 gm (4 ml) of

90 % formic acid

 Heat the reaction mixture on a water bath at 100 oC for 2 hours. Cool the content in flask
and add 10 % NaOH solution slowly, with constant stirring untill the reaction mixture is
just alkaline to litmus.

 Filter the crude benzimidazole & wash it with ice cold water, The yield of pure
benzimidazole, having m.p. 171-172oC& yield is 85 %.

Chemical Reaction:
NH2
NHCHO
N
NaOH
HCOOH
-H20
NH2 N
NH2 H

o-phenylenediamine benzimidazole
(108.14) (118.14)

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Experiment No:________ Date :

Type of Reaction: Simple Condensation reaction through intramolecular cyclization

Principle and Reaction Mechanism:

The two carban nitrogen bonds in benzimidazole when disconnected gives o-phenylene diamine
and formic acid. Therefore, synthesis of benzimidazole is effected simply heating the o-
phenylenediamine and formic acid to gether (condensation type of reaction).

H
N Disconnected NH2

N + HCOOH
NH2
Benzimidazole
o-phenylene diamine formic acid

The proposed reaction Mechanism illustrated as below:

H
NH2 O +
H
N
C -OH
H OH
NH2 O
NH2

o-phenylenediamine
(108.14)
-H+

H
H

N H
N H
Cyclization
OH O
N+
NH2
H
H

-OH

H
N

benzimidazole

KBIPER Medicinal Chemistry-I Page 6

Experiment No:________ Date :

Calculation:

M.W of o-phenylenediamine = 108.144 gm/mol

M.W of Benzimidazole = 118.14 g/mol
 Theoretical Yield
M.W of o-phenylenediamine = M.W of Benzimidazole

108.14 gm of o-phenylene diamine = 118.14 gm of Benzimidazole

6.7 gm of o-phenylene diamine = (?)

6.7 ×118.14

= 108.14
= 7.32 gm of benzimidazole
 Practical Yield of Benzimidazole_____________ (x gm)
 % yield:
= Practical yield X 100 / Theoretical yield
= _____________ X 100 / 7.32
= ___________% w/w (y % w/w)

 Observed Melting point of Benzimidazole _________ __(171-172oC)

Result:

Sr No.

1. Theoretical Yield Benzimidazole 7.32gm

2. Practical Yield Benzimidazole x gm

3. % Practical Yield Benzimidazole y%

4 Melting Point(oC) Benzimidazole (171-172oC)

5. Structure of Benzimidazole
H
N

Benzimidazole

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Experiment No:________ Date :

 Study Question:
1 In synthesis of benzimidazole which type of reaction occur.

2 In benzimidazole structure which heterocyclic ring is present.

3 Give the name of starting chemical in synthesis of benzimidazole.

4 Give the molecular formula of benzimidazole.

5 Give the molecular weight of benzimidazole

6 Give the use of benzimidazole.

7 Give the structure of benzimidazole.

KBIPER Medicinal Chemistry-I Page 8

Experiment No:________ Date :

Experiment No: 3
Aim: To synthesize benztriazole from o-phenylene diamine.
Reference: Furniss BS., Hannaford AJ., Smith PW., and Tatchell AR. In Vogel’s Text Book of
Practical Organic Chemistry; 5th Edn; Pearson Education Private Limited, New Delhi, 2005, pp
1163.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder

Reagents and Chemicals:

Sr. Chemical used Quantity required Total quantity required per
No. Batch of 30 students

1. o-phenylene diamine 5.4 gm 160 gm

2. Glacial acetic acid 6 gm(5.8 ml) 180 ml

3. Sodium nitrite 3.8 gm 120 gm

Procedure:

 Dissolve 5.4 gm of o-Phenylene diamine in a mixture of 6 gm (5.8 ml) of glacial acetic

acid and 15 ml of water in a 250 ml of beaker and slightly warm it

 Cool the clear solution to 15-20 C, stir the content in beaker magnetically or by glass rod
and then add solution of 3.8gm of sodium nitrite in 10ml of water in one portion

 Warm the reaction mixture for 3-4 minutes,untill the temprature of the reaction mixture
reaches around 80 C and then it begin to cool while the colour change from deep red to
pale brown. Continue stirring the mixture for 15 minutes,by which time the temperature
will have dropped to 35 C,and then throughly chill the content in flask in an ice water
bathfor half an hour

 So, pale brown colour solid separate out.

 Collect it, wash it with cold water and recrystalized the crude product with hot water using
charcoal as adsorbant.

 Chemical Reaction:
NH2
N
HNO2
N

NH2 N
H

o-phenylenediamine
benztrizole
(108.14)
(119.24)

KBIPER Medicinal Chemistry-I Page 9

Experiment No:________ Date :

Type of Reaction: Intra molecular cyclization reaction of monodiazonium salt

Principle and Reaction mechanism:

Disconnection approach for benztriazole gives monodiazonium salts of o-phenylene diamine,

which then converted in to o-phenylene diamine & nitrous acid. Therefore, benztriazole can be
prepared by treating o-phenylendiamine with nitrous acid (liberated during reaction between
sodium nitrite and acetic acid) to form monodiazonium salt that follow spontaneous intramolecular
cyclization reaction to give benztriazole.

The Proposed reaction mechanism illustrated as below:

H
NH2 N
N
HO N

OH
O
NH2 NH2

o-phenylenediamine
Cyclization

H
H
N
N
N
N OH
N
N
H

Calculation: benztrizole

M.W of o-phenylenediamine = 108.144 gm/mol

M.W of Benztriazole = 119.12 g/mol
 Theoretical Yield:
 M.W of o-phenylenediamine = M.W of Benztriazole

108.14 gm of o-phenylene diamine = 119.12 gm of Benztriazole

5 gm of o-phenylene diamine = (?)

= 5.4 ×119.12 / 108.14

= 5.95 gm of Benztriazole

KBIPER Medicinal Chemistry-I Page 10

Experiment No:________ Date :

 Practical Yield of Benztriazole ___________gm (x gm)

 % yield:

= Practical yield X 100 / Theoretical yield

= _____________ X 100 / 5.95

= ___________% w/w (y % w/w)

 Observed Melting point of Benztriazole_________ __(99-100oC)

Result:

Sr No.

1. Theoretical Yield Benztriazole 5.95 gm

2. Practical Yield Benztriazole x gm

3. % Practical Yield Benztriazole y%

4 Melting Point(oC) Benztriazole (99-100 o C)

5. Structure of Benztriazole
H
N
N
N

Benztriazole

KBIPER Medicinal Chemistry-I Page 11

Experiment No:________ Date :

 Study Question:
1 In synthesis of benztriazole which type of reaction occur.

2 In benztriazole structure which heterocyclic ring present.

3 Give the name of starting chemical in synthesis of benztriazole.

4 Give the molecular formula of benztriazole.

5 Give the molecular weight of benztriazole

6 Give the use of benztriazole.

7 Give the structure of benztriazole.

KBIPER Medicinal Chemistry-I Page 12

Experiment No:________ Date :

Experiment No: 4
Aim: To Synthesis 5, 5 –diphenyl hydantoin (Phenytion) from Benzil and urea
Reference: Furniss BS., Hannaford AJ., Smith PW., and Tatchell AR. In Vogel’s Text Book of
Practical Organic Chemistry; 5th Edn; Pearson Education Private Limited, New Delhi, 2005, pp
1153.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder
Reagents and Chemicals:

Sr. Chemical used Quantity required Total quantity required per

No. Batch of 30 students
1. Benzil 2gm 150gm
2. Urea 1.1gm 100gm
3. 30% NaOH 6ml 500ml
4. Ethanol 28ml 1200ml
5. Conc HCl q.s q.s

Procedure:
 Place 2 gm of benzil,1.1 gm of urea ,6 ml 30 % NaOH and 28 ml ethanol in 250 ml Round
bottom flask
 Attach reflux condenser and boil the reaction mixture under reflux for alleast 2 hrs, allow
to cool the reaction mixture to R.T and pour it into 150 ml of ice cold water and stir
thoroughly
 Allow to stand the mixture for 10-15 minutes, andfilter under suction to remove any
insoluble by product, make the filtrate strongly scidic with concHCl and allow to cool their
action mixture in ice bath and immediately filter off the obtained ppts under suction
 Recrystallition from ethanol to obtain pure 5,5-di phenyl hydantoin, having Melting Point
is 297-298oC

Type of reaction: Base Catalysed intramolecular cyclization with subsequent pinacolic

rearrangement reaction

Principle:
Disconnection approach in 5, 5-diphenyl hydantoin (Phenytoin) gives benzil and urea.
Thereforebase ctatalyse reaction between benzil & urea is used for synthesis of phenytoin. The
reaction is proceeding via base catalyzed intramolecular cyclization to form an intermediate
heterocyclic pinacol, which on acidification yield phenytoin(hydantoin) as a result of a pinacolic
rearrangement (1,2-diphenyl shift) reaction

Reaction mechanism: One of the NH2 group present in the urea, in presence of NaOH undergoes
nucleophilic attack on one of the carbonyl carban of benzil. Subsequently second amino(NH 2)
group of urea undergoes same reaction toward another carbonyl carban of benzil through

KBIPER Medicinal Chemistry-I Page 13

Experiment No:________ Date :

intramolecular cyclization reaction to gives diol type of heterocyclic ring,compounds containg

adjacent diol, when treated with acid undergoes rearrangement reaction to ketone via carbocation
intermediate with rearrangement similar to wagner-meerwein shift called pinacol rearrangement,
in similar fashion , acid protonated one of the hydroxyl group of diol type of heterocyclic ring and
converted in to hydronium ion , which is highly liable to give carbocation intermediate, &
thereafter phenyl group migrate with it’s bonded electrone towards electron deficient carbocation
to give protonated [Link] ketone is highly unstable form, therefore leave hydronium
ion to from 5,5-di phenyl hydantoin (phenytoin)

Chemical Reaction:
Ph O Ph
H2N H
N
Ph
NaOH/C2H5OH
O O

H2N N
Ph O H
O

benzil urea 5,5-diphenyl hydantoin

(phenytoin)

(86.09gm/mol) (252.27 gm/mol)

Reaction mechanism
Ph O OH
H2N H
N
Ph
NaOH/C2H5OH
O O
Ph
H2N N
Ph O H
OH
benzil urea
intermediate
Pinacol rearr., H+

OH2+
Ph H H
H + N N
N Ph Ph
Ph O -H2O O
O Ph Ph
+ N N
N H H
H OH OH
OH
1,2-Phenyl shift

Ph Ph
H H
N N
Ph Ph
-H+
O O

N N
H H
+
HO O
5,5-diphenyl hydantoin
Protonated ketone
(phenytoin)

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Experiment No:________ Date :

Use: Phenytoin is well known anti-epileptic drug.

Calculation

M.W of Benzil = 210.23 gm/mol

M.W of Phenytoin = 252.27 g/mol

 Theoretical Yield

M.W of Benzil = M.W of Phenytoin

210.23 gm of benzil = 252.27 gm of phenytoin

2 gm of benzil = (?)

2 × 252.27
= 86.09

= 5.87 gm of Phenytoin
 Practical Yield of Phenytoin ____________ gm (x gm)
 % Yield:

= Practical yield X 100 / Theoretical yield

= _____________ X 100 / 5.95

= ___________% w/w (y % w/w)

 Observed Melting point of Phenytoin __________ (297 oC)

 Result:

Sr No.

1. Theoretical Yield of 5.87 gm

Phenytoin

2. Practical Yield of x gm
Phenytoin

3. % Practical Yield of Y%
Phenytoin

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Experiment No:________ Date :

4 Melting Point(oC) of (297oC)

Phenytoin

5. Structure of Phenytoin
Ph
H
N
Ph
O

N
H
O

KBIPER Medicinal Chemistry-I Page 16

Experiment No:________ Date :

 Study Question:

1 In synthesize of Phenytion which type of reaction occur.

2 Phenytion comes under which class.

3 Give the name of starting chemical in synthesis of Phenytion.

4 Give the molecular formula of Phenytion.

5 Give the molecular weight of Phenytion.

6 Give the use of Phenytion.

7 Give the structure of Phenytion.

8 What is anti convlsant.

KBIPER Medicinal Chemistry-I Page 17

Experiment No:________ Date :

Experiment No: 5
Aim: To carry out synthesis of benzocaine from p-aminobenzoic acid.
Reference: Furniss BS., Hannaford AJ., Smith PW., and Tatchell AR. Vogel’s Text Book of
Practical Organic Chemistry; 5th Edn; Pearson Education Private Limited, New Delhi, 2005, pp
701.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder
Reagents and Chemicals: p-amino benzoic acid, absolute ethanol, dry hydrogen chloride,
sodium carbonate, rectified spirit

Principle: The synthesis of benzocaine is based on esterification of p-amino benzoic acid. P-amino
benzoic acid whenever treated with absolute ethanol in presence of dry HCl, it will be converted
in to ethyl-p-amino benzoate, which is known as benzocaine.
Reaction:

Procedure:
 Take 8 ml absolute ethanol in 250 ml RBF and saturate it with dry HCl. Add 1.2 gm p-
amino benzoic acid in to it. Reflux the reaction mixture for 2 hours. White solid ppts will
be separated out. Add hot water in to it till the solid mass dissolves and saturate it with
sodium carbonate.
 Crude precipitates of benzocaine will be separated out. Filter the precipitates.
 Dry the product and weigh it. Report Theoretical yield, Practical yield, %Practical yield
and Melting point of crude and standard. Recrystallize from rectified spirit, m.p. 91 °C

Calculation:
M.W. of p-amino benzoic acid = 137 gm/mol
M.W. of benzocaine = 165 gm/mol

 Theoretical yield:
M.W. of p-amino benzoic acid = M.W. of benzocaine
137 gm of p-amino benzoic acid = 165 gm of benzocaine
1.2 gm of p-amino benzoic acid = (?)
= 165 X 9.5/ 137
= 1.45 gm of benzocaine

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Experiment No:________ Date :

 Practical yield = ___________ gm (x gm)

 % yield
= Practical yield X 100 / Theoretical yield
= x X 100 / 1.45
= ____________ % w/w (y % w/w)
Result:

Sr No.

1. Theoretical Yield of 1.45 gm

benzocaine

2. Practical Yield of x gm
benzocaine

3. % Practical Yield of y%
benzocaine

4 Melting Point (oC) of (91 oC)

benzocaine

5. Structure of benzocaine

KBIPER Medicinal Chemistry-I Page 19

Experiment No:________ Date :

 Study Question:
1 Write the principle of benzocaine.

2 Benzocaine comes under which class.

3 Give the name of starting chemical in synthesis of benzocaine.

4 Give the molecular formula of benzocaine.

5 Give the molecular weight of benzocaine.

6 Give the use of benzocaine.

7 Give the structure of benzocaine.

8 What is local anesthetic.

KBIPER Medicinal Chemistry-I Page 20

Experiment No:________ Date :

Experiment No: 6
Aim: To carry out synthesis of 2,3-diphenylquinoxaline from o-phenylene diamine.
Reference: Furniss BS., Hannaford AJ., Smith PW., and Tatchell AR. Vogel’s Text Book of
Practical Organic Chemistry; 5th Edn; Pearson Education Private Limited, New Delhi, 2005, pp
1190.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder
Reagents and Chemicals: benzil, o-phenylene diamine, rectified spirit

Principle: The synthesis of 2,3-diphenylquinoxaline is based on heterocyclic condensation. Benzil

whenever treated with o-phenylene diamine, both molecules will be cyclic condensed and will be
converted in to heterocyclic nucleus known as quinoxaline, which is ten membered heterocyclic
ring and two nitrogens are present as a heteroatom at 1 and 4 position. The formed compound will
be known as 2,3-diphenylquinoxaline.
Reaction:

Procedure:
 Take 8 ml rectified spirit in conical flask. Add 2.1 gm benzil in to it and slight warm the
reaction mixture. Add 1.1. gm o-phenylene diamine in to it.
 Warm the reaction mixture on water bath for 30 minutes.
 After heating add apprx. 20 ml water so that reaction mixture becomes slightly cloudiness.
Allow the mixture to cool
 Crude precipitates of 2,3-diphenylquinoxaline will be separated out. Filter the precipitates.
 Dry the product and weigh it. Report Theoretical yield, Practical yield, %Practical yield
and Melting point of crude and standard. Recrystallize from rectified spirit, m.p. 125-126
°C

Calculation:
M.W. of benzil = 210 gm/mol
M.W. of 2,3-diphenylquinoxaline = 282 gm/mol

 Theoretical yield:
M.W. of benzil = M.W. of 2,3-diphenylquinoxaline
210 gm of benzil = 282 gm of 2,3-diphenylquinoxaline

KBIPER Medicinal Chemistry-I Page 21

Experiment No:________ Date :

2.1 gm of benzil = (?)

= 282 X 2.1/ 210
= 2.82 gm of 2,3-diphenylquinoxaline
 Practical yield = x gm
 % yield
= Practical yield X 100 / Theoretical yield
= _________X 100 / 2.82
= __________% w/w ( y % w/w)
Result:

Sr No.

1. Theoretical Yield of 2,3- 2.82 gm

diphenylquinoxaline

2. Practical Yield of 2,3- X gm

diphenylquinoxaline

3. % Practical Yield of 2,3- Y%

diphenylquinoxaline

4 Melting Point(oC) of 2,3- (125-126 oC)

diphenylquinoxaline

5. Structure of 2,3-
diphenylquinoxaline

KBIPER Medicinal Chemistry-I Page 22

Experiment No:________ Date :

 Study Question:
1 Write the principle of 2,3-diphenylquinoxaline.

2 Which heterocyclic ring present in 2,3-diphenylquinoxaline.

3 Give the name of starting chemical in synthesis of 2,3-diphenylquinoxaline.

4 Give the molecular formula of 2,3-diphenylquinoxaline.

5 Give the molecular weight of 2,3-diphenylquinoxaline.

6 Give the use of 2,3-diphenylquinoxaline.

7 Give the structure of 2,3-diphenylquinoxaline.

KBIPER Medicinal Chemistry-I Page 23

Experiment No:________ Date :

Experiment No: 7
Aim: To Synthesis phenothiazine from diphenylamine.
Reference: Cuming WM., In Systemic Organic Chemistry. 1937, 325-326.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder
Reagents and Chemicals: Diphenylamine, anhydrous AlCl3, sulfur

Procedure:
Take 5.5 gm of 2.75 gm of diphenylamine, 1 gm sulfur and 0.5 gm anhydrous aluminium
trichloride in conical flask. Heat the reaction mixture. The reaction mixture will melt and sets at
140 oC with rapid evolution of hydrogen sulfide. Slight cool the mixture, the mixture will be
slackened. Again heat the mixture up to 160 oC. The mixture is allowed to cool. Extract the product
first with water and second with dilute alcohol. The residue is pure phenothiazine. Recrystallise it
from alcohol. Report the yield.

Reaction:

Calculation:

M.W. of phenothiazine = 169 gm/mol

M.W. of diphenylamine = 199 gm/mol

 Theoretical Yield
M.W. of phenothiazine = M.W. of diphenylamine

169 gm of phenothiazine = 199 gm of diphenylamine

2.75 gm of phenothiazine = (?)

2.75×199
= 169

= 3.24 gm of diphenylamine
 Practical Yield of phenothiazine ________ gm (x gm)

KBIPER Medicinal Chemistry-I Page 24

Experiment No:________ Date :

 % yield:

= Practical yield X 100 / Theoretical yield

= __________ X 100 / 2.82

= ____________ % w/w (y% w/w)

 Observed Melting point of phenothiazine ________________(185oC)

Result:

Sr Content Observation
No.

1. Theoretic Yield of 3.24 gm

phenothiazine

2. Practical Yield of X gm
phenothiazine

3. % Practical Yield of Y%
phenothiazine

4 Melting Point(oC) of (185oC)

phenothiazine

5. Structure of
phenothiazine

KBIPER Medicinal Chemistry-I Page 25

Experiment No:________ Date :

 Study Question:
1 Write the principle of phenothiazine.

2 How many ring is present in phenothiazine?

3 Give the name of starting chemical in synthesis of phenothiazine.

4 Give the molecular formula of phenothiazine.

5 Give the molecular weight of phenothiazine.

6 Give the use of phenothiazine

7 Give the structure of phenothiazine.

8 What is antipsychotic drug.

KBIPER Medicinal Chemistry-I Page 26

Experiment No:________ Date :

Experiment No: 8
Aim: To Synthesis 1,3-diphenylpyrazole from diphenyl hydrazone and ethylene glycol.
Apparatus: Beaker, conical flask, glass rod, funnel, 10 ml measuring cylinder
Reagents and Chemicals: Diphenyl hydrazone, ethylene glycol, ferric chloride, tert-butyl
hydroproxide, acetylacetone, NaCl, ethylacetate, sodium acetate.

Reaction:

HO
N
N N

HO
HN
Ethane-1,2-diol
5 % FeCl3
t-Butylhydroperoxide
90o-120o C, 6 hr
Diphenylhydrazone 1,3-Diphenyl-1-H-pyrazole

Procedure:
Take 0.91 gm diphenylhydrazone and dissolve it in 5 ml ethylene glycol and 5% ferric chloride
solution. Prepare a solution of tert-butylhydroperoxide (1.06 gm) in 5 ml of acetylacetone kept a
temp ranging from 90 to 100 oC. The whole mixture is left to reach room temp and is kept under
stirring at this temp for 6 hrs.

At the end of the reaction, the mixture is poured in to water and extracted three times with
ethylacetate. The joined organic phases are washed with water, then with a saturated solution of
NaCl and finally anhydrified with Na2SO4 and evaporated under vacuum.

Calculation:

M.W. of diphenyl hydrazone = 196 gm/mol

M.W. of 1,3-diphenylpyrazole = 220 gm/mol

 Theoretical Yield

196 gm of diphenyl hydrazone = 220 gm of 1,3-diphenylpyrazole

0.91 gm of diphenyl hydrazone = (?)

KBIPER Medicinal Chemistry-I Page 27

Experiment No:________ Date :

0.91×220
= 196

= 1.02 gm of 1,3-diphenylpyrazole
 Practical Yield of 1,3-diphenylpyrazole _________ gm (x gm)
 % Yield:

= Practical yield X 100 / Theoretical yield

= __________ X 100 / 2.82

= ____________ % w/w (y% w/w)

 Observed Melting point of 1,3-diphenylpyrazole _________ (185oC)

Result:

Sr Content Observation
No.

1. Theoretic Yield of 1,3- 1.02 gm

diphenylpyrazole

2. Practical Yield of 1,3- X gm

diphenylpyrazole

3. % Practical Yield of 1,3- Y%

diphenylpyrazole

4 Melting Point(oC) of
1,3-diphenylpyrazole

KBIPER Medicinal Chemistry-I Page 28

Experiment No:________ Date :

5. Structure of 1,3-
diphenylpyrazole

KBIPER Medicinal Chemistry-I Page 29

Experiment No:________ Date :

 Study Question:
1 What is analgesic and antipyreticdrug.

2 Which heterocyclic ring is present in diphenylpyrazole?

3 Give the name of starting chemical in synthesis of diphenylpyrazole.

4 Give the molecular formula of diphenylpyrazole.

5 Give the molecular weight of diphenylpyrazole.

6 Give the use of diphenylpyrazole.

7 Give the structure of diphenylpyrazole.

KBIPER Medicinal Chemistry-I Page 30

Experiment No:________ Date :

Experiment No: 9
Aim: To perform assay of aspirin as per IP2014.

Reference: Indian Pharmacopoeia 2014. Government of India, Ministry of Health & Family
Welfare. The Indian Pharmacopoeia Commission, Ghaziabad, Vol II, pp 1093.
Apparatus: Volumetric flask, measuring cylinder, analytical balance, weight box, beaker, burette
and conical flask.
Reagents and chemicals: aspirin tablets, 0.5 M H2SO4 solution, 0.05 M bromine solution
Procedure:
Weigh and powder 20 tablets. Weigh a quantity of the powder containing about 0.3 gm of aspirin,
dissolve in 10 ml of 0.5 M H2SO4 solution and boil under a reflux condenser for 1 hour. Cool,
transfer to a separating funnel with the aid of small quantities of water, and extract the liberated
salicylic acid with four quantities, each of 20 ml of ether. Wash the combined ether extracts with
two quantities, each of 5 ml of ether. Remove the ether in a current of air at a temperature not
exceeding30 oC. Dissolve the residue in 20 ml of 0.5 M NaOH and dilute to 200 ml with water.
Transfer 50 ml to a stoppered flask, add 50 ml of 0.05 M bromine and 5 ml HCl, protect the mixture
from light and shake repeatedly during 25 minutes. Add 25 ml of KI solution. Shake thoroughly
and titrate with 0.1 M sodium thiosulphate using starch solution, added towards the end point of
the titration, as an indicator
Factor: Each ml of 0.5M sodium hydroxide is required to 0.04504 of Aspirin.

FOR THE ASSAY OF ASPIRIN

Observation:
 Burette: 0.1 M sodium thiosulphate
 Conical flask: 50 ml of solution (0.3 gm aspirin+ 10 ml of 0.5 M H2SO4 + 20 ml
ether+ 20 ml of 0.5 M NaOH + 200 ml water) + 50 ml of 0.05 M bromine +5 ml
HCl
 Color change:
 Indicator: starch solution
Observation table:
BURETTE I (ml) II (ml) Average
READING
FINAL

INITIAL

DIFFERENCE

KBIPER Medicinal Chemistry-I Page 31

Experiment No:________ Date :

Each ml 0.05 M bromine = 0.003003 gm Aspirin

_______ X ml of 0.05 M bromine = ?

= X ml * 0.05 * 0.003003 /1 * 0.05

= __________ gm of aspirin

Percentage of aspirin

3 gm sample contains = Y gm of aspirin

100 gm sample contains =?

= 100* Y/ 3

= z % w/w of aspirin

Result: The given sample of aspirin contains __________% w/w of aspirin as per IP 2014.

__________________________OR____________________________

Aim: To perform assay of aspirin.

Apparatus: Burette, Pipette, funnel, conical flask, beaker.
Reagents and chemicals: 0.1 N HCl, 0.1 N NaOH, Phenol red, methyl orange, Na2CO3
Principle: Aspirin assay is based on acid base back titration. Aspirin is insoluble in water and
weak acid. So direct titration is not possible. Aspirin reacts with excess of NaOH on boiling which
convert aspirin to sodium salicylate. Then unreacted NaOH remaining after neutralization of
aspirin is titrated with HCl and back reading is taken. Blank reading without aspirin is taken. And
difference between these two readings give amount of NaOH require for aspirin neutralization.
Reaction
COOH COOH

OCOCH3 OH

H2O CH3COOH
(NaOH)

Aspirin Salycilic Acid

KBIPER Medicinal Chemistry-I Page 32

Experiment No:________ Date :

COOH COONa

OH OH

CH3COOH 2NaOH CH3COONa 2H2O

Sodium Acetate
Sodium Salycilate

PROCEDURE
PROCEDURE FOR STANDARDIZATION OF HCL:
Weigh accurately 100 mg of anhydrous Na2CO3 in Conical flask. Add 20 ml distil water. Shake it
properly. Add 2-3 drops of Methyl red as an indicator and titrate it against HCl. Find out end point,
take 2 readings and calculate normality of HCl.

PROCEDURE:
Weigh accurately 100 mg of sample. Add 25 ml of 0.1 N NaOH. Boil gently for 10 min. cool and
titrate excess alkali with 0.1 N HCl using phenol red as indicator. Take back reading. Repeat
procedure without taking sample and note down as blank reading. Difference between these two
readings represent amount of NaOH require to neutralize Aspirin.

Each ml of 0.1 N HCL equivalent to 0.0009 gm of Aspirin

RESULT:

1. Normality of HCl = ___________

2. Blank reading _________ ml of 0.1 N HCl requires to neutralize total NaOH.
3. Back reading _________ ml of 0.1 N HCl require neutralizing unreacted NaOH.
4. Blank reading – Back reading ________ ml of 0.1 N NaOH requires to neutralize
reacted aspirin.
5. Assay purity ______________.
6. The given sample of aspirin Complies or Does not Comply with requirement of assay
purity as per IP.

STADARDIZATION OF HCL
Na2CO3 + 2HCl H2CO3 + 2NaCl

KBIPER Medicinal Chemistry-I Page 33

Experiment No:________ Date :

Observation:
 Burette : 0.1N HCl Solution
 Conical flask: anhydrous Na2CO3 + 20 ml water + methyl Orange indicator
 Color change: yellow to orange red
 Indicator : methyl orange

Observation table:-
BURETTE I (ml) II (ml) Average
READING

FINAL

INITIAL

DIFFERENCE

Calculation for normality

Normality= Wt of primary std compound

Equivalent wt * Burette reading

FOR THE ASSAY OF ASPIRIN

Observation:
 Burette : 0.1 N HCl Solution
 Conical flask: 150 mg aspirin + 25 ml NaOH + phenol red indicator
 Color change: pink to yellow
 Indicator: Phenol red

KBIPER Medicinal Chemistry-I Page 34

Experiment No:________ Date :

Observation table:-
BURETTE I (ml) II (ml) Average
READING

FINAL

INITIAL

DIFFERENCE

Blank reading=----------------ml

Actual reading= Blank- Back

= --------ml

Calculation:

Each ml 0.1 N NaOH = 0.009 gm Aspirin

X ml of 0.1 N NaOH = ?

= X ml * 0.1 * 0.009 /1 * 0.1

= Y gm of aspirin

= Y* 1000 mg aspirin

Percentage of aspirin

100 mg sample contains = Y mg of aspirin

100 mg sample contains = ?

= 100* Y/ 100

= ____________ % w/w of aspirin (z % w/w of aspirin)

Result: The given sample of aspirin contains __________% w/w of aspirin.

KBIPER Medicinal Chemistry-I Page 35

Experiment No:________ Date :

 Study Question:
1 What is analgesic and antipyretic drug.

2 Which type of reaction occur in assay of aspirin.

3 What indicator use in assay of aspirin.

4 Give the molecular formula of aspirin.

5 Give the molecular weight of aspirin.

6 Give the use of aspirin.

7 Aspirin comes under which class.

8 Give the structure of aspirin

KBIPER Medicinal Chemistry-I Page 36

Experiment No:________ Date :

Experiment No: 10
Aim: To perform assay of Chlorpromazine as per IP2014.
Reference: Indian Pharmacopoeia 2014. Government of India, Ministry of Health & Family
Welfare. The Indian Pharmacopoeia Commission, Ghaziabad, Vol II, pp 1378.
Apparatus: Volumetric flask, measuring cylinder, analytical balance, weight box, beaker, burette
and conical flask.
Reagents and chemicals: Chlorpromazine tablets, dilute hydrochloric acid
Principle
Chlorpromazine is chromophoric group containing drug. It can be analyzed by UV
spectrophotometer by using A= abc equation.
Procedure:
Weigh and powder 20 tablets. Weigh accurately a quantity of the powder containing about 0.1 gm
of chlorpromazine hydrochloride, add 5 ml of dilute hydrochloric acid and 200 ml of water. Shake
for 15 minutes and add sufficient water to produce 500 ml. Centrifuge about 15 ml and to 5 ml of
the clear, supernatant liquid. Add 10 ml of dilute hydrochloric acid and sufficient water to produce
200 ml. Measure the absorbance of the resulting solution at the maximum at about 254 nm.
Calculate the content of C17H19ClN2S,HCl, taking 915 as the specific absorbance at 254 nm.

Calculation
A= abc
Where A= absorbance, a= absorptivity, b= path length, c= concentration
a= 915 (1%,1 cm), b=1 cm, A=? c=? (gm/100 ml)

RESULT:

1. Avg. weight of 20 tablets is________________mg.

2. The given tablet ____________________with requirement of uniformity of weight test as
per IP 2007.
3. Absorbance of test solution at 275 nm is _______________.
4. Assay purity is ___________
5. Content of active ingredient is ______________.
6. The given tablet ______________ with test of content uniformity as per IP 2014.

KBIPER Medicinal Chemistry-I Page 37

Experiment No:________ Date :

WEIGHT UNIFORMITY TEST

Sr. Weight of tablet Sr. Weight of tablet

no
no

1 11

2 12

3 13

4 14

5 15

6 16

7 17

8 18

9 19

10 20

Avg weight of 20 tablet =

% deviation =

% limit range =

Range in mg =

No of tablet deviated =

KBIPER Medicinal Chemistry-I Page 38

Experiment No:________ Date :

__________________________________OR_______________________________________________

Aim: To perform assay of Chlorpromazine.

Reference: Soliman SA., Abdine H., and Zakhari NA, “Chemistry of nonaqueous titration of
chlorpromazine.” J. Pharm. Sci. 1975, 64(1), 129-132.
Apparatus: Volumetric flask, measuring cylinder, analytical balance, weight box, beaker,
burette and conical flask.
Reagents and chemicals: Chlorpromazine tablets, perchloric acid, glacial acetic acid, acetone,
acetic anhydride, mercuric acetate solution
Principle
It is a non-aqueous titration. Chlorpromazine hydrochloride is very weakly basic to react
quantitatively with perchloric acid. So mercuric acetate is added which replace the halide ion with
an equivalent quantity of acetate ion, which is strong base in acetic acid.
PROCEDURE
Preparation of perchloric caid
Mix 8.5 ml of perchloric acid with 500 ml of anhydrous glacial acetic acid and 25 ml of acetic
anhydride, cool and add anhydrous glacial acetic acid to produce 1000 ml.

Standardization of perchloric acid

Weigh accurately about 0.35 gm of potassium hydrogen phthalate and dissolve it in 50 ml of
anhydrous glacial acetic acid. Add 0.1 ml of crystal violet solution and titrate with the perchloric
acid solution until the violet color changes to emerald green. Perform a blank determination.

Procedure
Weigh accurately about 0.6 gm and dissolve in 200 ml of acetone. Add 15 ml of mercuric acetate
solution. Titrate with 0.1 M perchloric acid, using a saturated solution of methyl orange in acetone
as indicator. Perform a blank determination and make a necessary correlation.

Each ml of 0.1 M perchloric acid equivalent to 0.03553 gm of C17H19ClN2S,HCl

Mercuric acid solution: A 5 % w/v solution of mercuric acetate in glacial acetic acid solution.

KBIPER Medicinal Chemistry-I Page 39

Experiment No:________ Date :

Observation:
 Burette: 0.1 M Perchloric acid
 Conical flask: 0.6 gm chlorpromazine powder+ 15 ml mercuric acetate+ methyl
orange in acetone
 Color change:
 Indicator: methyl orange in acetone
Observation table: -
BURETTE I (ml) II (ml) Average
READING

FINAL

INITIAL

DIFFERENCE

Calculation:

Each ml of 0.1 M perchloric acid equivalent to 0.03553 gm of C17H19ClN2S,HCl

X ml of 0.1 M perchloric acid is equivalent to = ?

= ___________X ml * 0.1 * 0.03553 /1 * 0.1

= _____________Y gm of C17H19ClN2S, HCl

Percentage of C17H19ClN2S, HCl

0.6 gm sample contains = Y gm of C17H19ClN2S,HCl

100 gm sample contains = ?

= 100* Y/ 0.6

KBIPER Medicinal Chemistry-I Page 40

Experiment No:________ Date :

Result:

The given sample contains ____________ % w/w of chlorpromazine.

KBIPER Medicinal Chemistry-I Page 41

Experiment No:________ Date :

 Study Question:
1 What is antipsychotic drug.

2 Which type of reaction occur in assay of chlorpromazine.

3 What indicator use in assay of chlorpromazine.

4 Give the molecular formula of chlorpromazine.

5 Give the molecular weight of chlorpromazine.

6 Give the use of chlorpromazine.

7 Chlorpromazine comes under which class.

8 Give the structure of chlorpromazine.

KBIPER Medicinal Chemistry-I Page 42

Experiment No:________ Date :

Experiment No: 11
Aim: To perform assay of phenobarbitone sodium as per IP 2014.
Reference: Indian Pharmacopoeia 2014. Government of India, Ministry of Health & Family
Welfare. The Indian Pharmacopoeia Commission, Ghaziabad, Vol III, pp 2470.
Apparatus: Volumetric flask, measuring cylinder, analytical balance, weight box, beaker,
burette and conical flask, potentiometer.
Reagents and chemicals: phenobarbitone sodium tablets, NaOH solution, NaCl, HCl,
Principle:
Potentiometry is one of the volumetric techniques of electro analytical chemistry. It involves the
measurement of potential difference between an indicator electrode and a reference electrode.
Reaction such as redox, precipitation and neutralization involving marked change in potential at
the end point can be carried out on potentiometer. It helps in exact location of the end point.

Reference electrode: Saturated calomel electrode

Indicator electrode: Glass electrode
The end point can be found potentiometrically depends upon the magnitude of change in E.M.F.
near to equivalence point.

Procedure:
Weigh and powder 20 tablets. Weigh accurately a quantity of the powder containing about 0.15
gm of phenobarbitone sodium powder and add 2 ml of water and add 8 ml of 0.05 M Sulphuric
acid. Heat to boiling and cool. Add 30 ml of methanol and shake until dissolution is complete.
Titrate with 0.1 M NaOH solution, determine the end point potentiometrically. After the first
inflection, stop the addition of NaOH solution, add 10 ml of pyridine. Mix and continue the
titration until the second inflection is reached. The difference between the volumes represents the
amount of sodium hydroxide required.

KBIPER Medicinal Chemistry-I Page 43

Experiment No:________ Date :

WEIGHT UNIFORMITY TEST

Sr. Weight of tablet Sr. Weight of tablet

no
no

1 11

2 12

3 13

4 14

5 15

6 16

7 17

8 18

9 19

10 20

Avg weight of 20 tablet =

% deviation =

% limit range =

Range in mg =

No of tablet deviated =

Observation Table:

KBIPER Medicinal Chemistry-I Page 44

Experiment No:________ Date :

Sr no. ml of titrant Potential

Factor
1 ml of 0.1 M NaOH is equivalent to 0.02542 gm of C12H11N2NaO3
Result:
1) ml of 0.1 M NaOH required to neutralize phenobarbitone =________
2) mg of phenobarbitone =_________
3) % Assay purity=________
4) The given tablet does’t complies /complies with assay as per IP 2014

KBIPER Medicinal Chemistry-I Page 45

Experiment No:________ Date :

__________________________________OR_________________________

Aim: To perform assay of phenobarbitone.

Reference: A Practical book of Medicinal Chemistry by Dr Abhishek Tiwari, Nirali Prakashan
anpp 62.
Apparatus and instrument: Volumetric flask, measuring cylinder, analytical balance, weight
box, beaker, burette and conical flask
Reagents and chemicals: phenobarbitone. tablets, pyridine, thymophthalein, .
Procedure
Weigh accurately about 0.1g phenobarbitone, dissolve in 5ml of pyridine add 0.25ml of
thymophthalein solution and 10 ml of silver nitrate pyridine reagent.
Titrate with 0.1 M methanolic sodium hydroxide until a pure blue colour is obtained.
Repeat the operation without drug under examination.
The difference between the titration represents the amount of sodium hydroxide required.

Each 1 ml 0.1 M methanolic sodium hydroxide equivalent to 0.01161 gm of phenobarbitone.

KBIPER Medicinal Chemistry-I Page 46

Experiment No:________ Date :

 Study Question:
1 What is anticonvulsant drug.

2 Which type of reaction occur in assay of phenobarbitone.

3 What indicator use in assay of phenobarbitone.

4 Give the molecular formula of phenobarbitone.

5 Give the molecular weight of phenobarbitone.

6 Give the use of phenobarbitone.

7 Phenobarbitone comes under which class.

8 Give the structure of phenobarbitone

KBIPER Medicinal Chemistry-I Page 47

Experiment No:________ Date :

Experiment No: 12
Aim: To perform assay of atropine.
Apparatus: Volumetric flask, measuring cylinder, analytical balance, weight box, beaker, burette
and conical flask.
Reagents and chemicals:
Perchloric acid, glacial acetic acid, acetone, acetic anhydride, atropine sulfate
Principle:
Potentiometry is one of the volumetric techniques of electro analytical chemistry. It involves the
measurement of potential difference between an indicator electrode and a reference electrode.
Reaction such as redox, precipitation and neutralization involving marked change in potential at
the end point can be carried out on potentiometer. It helps in exact location of the end point.

Reference electrode: Saturated calomel electrode

Indicator electrode: Glass electrode
The end point can be found potentiometrically depends upon the magnitude of change in E.M.F.
near to equivalence point.

PROCEDURE
Preparation of perchloric caid
Mix 8.5 ml of perchloric acid with 500 ml of anhydrous glacial acetic acid and 25 ml of acetic
anhydride, cool and add anhydrous glacial acetic acid to produce 1000 ml.
Standardization of perchloric acid
Weigh accurately about 0.35 gm of potassium hydrogen phthalate and dissolve it in 50 ml of
anhydrous glacial acetic acid. Add 0.1 ml of crystal violet solution and titrate with the perchloric
acid solution until the violet color changes to emerald green. Perform a blank determination.

KBIPER Medicinal Chemistry-I Page 48

Experiment No:________ Date :

Procedure
Weigh accurately about 0.5 gm and dissolve in 30 ml of anhydrous glacial acetic acid and carry
out for nonaqueous titration determining the end point potentiometrically. Dissolve the prescribed
quantity of the substance being examined in a suitable volume of anhydrous glacial acid, warming
and cooling if necessary or prepare a solution as directed in the monograph and determine the
equivalence point potentiometrically using 0.1 M perchloric acid as titrant, unless otherwise
specified in the monograph potentiometric titration may be carried out using a glass electrode and
standard reference electrode, ex. Calomel reference electrode containing saturated solution of KCl
in water. Potentiometric titrations may also be carried out by using a glass electrode and a saturated
solution of KCl in water has been replaced by a saturated solution of KCl in methanol. It must be
ensured that no leakage of salt bridge solution occurs. Alternatively a combined electrode may be
used. The junction between the calomel electrode and the titration liquid should have a low
electrical resistance and there should be minimum of transfer of liquid from one side to other.
Perform a blank determination and make any necessary correction.
Each ml of 0.1 M perchloric acid is equivalent to 0.06768 gm of C17H23NO3, H2SO4.
Observation Table:

Sr no. ml of titrant Potential

KBIPER Medicinal Chemistry-I Page 49

Experiment No:________ Date :

Factor
1 ml of 0.1 M perchloric acid is equivalent to 0.02542 gm of C17H23NO3, H2SO4
Result:
1) ml of 0.1 M perchloric acid required to neutralize atropine =________
2) mg of atropine =_________
3) % Assay purity=________
4) The given tablet does’t complies /complies with assay.

KBIPER Medicinal Chemistry-I Page 50

Experiment No:________ Date :

 Study Question:
1 What is anticholinergic drug.

2 Which type of reaction occur in assay of atropine.

3 What indicator use in assay of atropine.

4 Give the molecular formula of atropine.

5 Give the molecular weight of atropine.

6 Give the use of atropine.

7 Atropine comes under which class.

8 Give the structure of atropine.

KBIPER Medicinal Chemistry-I Page 51

Experiment No:________ Date :

Experiment No: 13
Aim: To perform assay of ibuprofen as per IP2014.
Reference: Indian Pharmacopoeia 2014. Government of India, Ministry of Health & Family
Welfare. The Indian Pharmacopoeia Commission, Ghaziabad, Vol II, pp 1941.
Apparatus: Volumetric flask, measuring cylinder, analytical balance, weight box, beaker, burette
and conical flask.
Reagents and chemicals: Ibuprofen tablets, chloroform, ethanol (95 per cent), phenolphthalein
solution
Principle: Ibuprofen is a non-steroidal anti-inflammatory drug (NSAID) derived from propionic
acid and it is considered the first of the propionics. Ibuprofen is an acidic drug that can be
determined by neutralization titration by titrated against NaOH as a titrant and using neutral
phenolphthalein solution as an indicator.
Procedure
Weigh and powder 20 tablets. Weigh accurately a quantity of the powder containing about 0.5 gm
of ibuprofen, extract with 60 ml of chloroform for 15 minutes and filter through a sintered glass
crucible of porosity 3. Wash the residue with three quantities, each of 10 ml, of chloroform and
gently evaporate the filtrate just to dryness in a current of air. Dissolve the residue in 100 ml of
ethanol (95 per cent), previously neutralized to phenolphthalein solution, and titrate with 0.1 M
NaOH using phenolphthalein as indicator.
1 ml of 0.1 M NaOH is equivalent to 0.02063 gm of C13 H18O2.
WEIGHT UNIFORMITY TEST

Sr. Weight of tablet Sr. Weight of tablet

no
no

1 11

2 12

3 13

4 14

KBIPER Medicinal Chemistry-I Page 52

Experiment No:________ Date :

5 15

6 16

7 17

8 18

9 19

10 20

Avg weight of 20 tablet =

% deviation =

% limit range =

Range in mg =

No of tablet deviated =

Observation:
 Burette : 0.1 M NaOH
 Conical flask: 0.5 gm of ibuprofen +100 ml of ethanol (95 per cent) +
phenolphthalein solution
 Color change:
 Indicator : phenolphthalein solution
Observation table:-
BURETTE I (ml) II (ml) Average
READING

FINAL

INITIAL

DIFFERENCE

KBIPER Medicinal Chemistry-I Page 53

Experiment No:________ Date :

Calculation:

Each ml of 0.1 M NaOH is equivalent to 0.02063 gm of C13 H18O2.

X ml of 0.1 M NaOH is equivalent to = ?

= X ml * 0.1 * 0.02063 /1 * 0.1

= Y gm of C13 H18O2

Percentage of C13 H18O2

0.5 gm sample contains = Y gm of C13 H18O2

100 gm sample contains = ?

= 100* Y/ 0.5

Result:

The given sample contains ____________ % w/w of Ibuprofen.

KBIPER Medicinal Chemistry-I Page 54

Experiment No:________ Date :

 Study Question:
1 What is NSAID.

2 Which type of reaction occur in assay of ibuprofen.

3 What indicator use in assay of ibuprofen.

4 Give the molecular formula of ibuprofen.

5 Give the molecular weight of ibuprofen.

6 Give the use of ibuprofen.

7 Ibuprofen comes under which class.

8 Give the structure of ibuprofen.

KBIPER Medicinal Chemistry-I Page 55

Experiment No:________ Date :

Experiment No: 14

Aim: To perform assay of furosemide as per IP2014.

Reference: Indian Pharmacopoeia 2014. Government of India, Ministry of Health & Family
Welfare. The Indian Pharmacopoeia Commission, Ghaziabad, Vol II, pp 1833.
Apparatus and instrument: Volumetric flask, measuring cylinder, analytical balance, weight
box, beaker, burette and conical flask, specterophotometer.
Reagents and chemicals: Furosemide tablets, 0.1 M sodium hydroxide solution
Principle
Furosemide is chromophoric group containing drug. It can be analyzed by UV spectrophotometer
by using A= abc equation.
Procedure
Weigh and powder 20 tablets. Weigh a quantity of the powder containing 0.1 gm of Furosemide
and shake with 150 ml of 0.1 M sodium hydroxide for 10 minutes. Add sufficient 0.1 M sodium
hydroxide to produce 250 ml and filter. Dilute 5 ml to 200 ml with 0.1 M sodium hydroxide and
measure the absorbance of the resulting solution at the maximum at about 271 nm (2.4.7).
calculate the content of C12H13ClN2O5S taking 580 as the specific absorbance at 271 nm.

WEIGHT UNIFORMITY TEST

Sr. Weight of tablet Sr. Weight of tablet

no
no

1 11

2 12

3 13

4 14

5 15

6 16

KBIPER Medicinal Chemistry-I Page 56

Experiment No:________ Date :

7 17

8 18

9 19

10 20

Avg weight of 20 tablet =

% deviation =

% limit range =

Range in mg =

No of tablet deviated =

Calculation:

A= abc

Where A= absorbance, a= absorptivity, b= path length, c= concentration

a= 915 (1%,1 cm), b=1 cm, A=? c=? (gm/100 ml)

Result:
The percentage purity of chlorapromazine tablet is found to be __________% w/w furosemide as
per IP 2014.

KBIPER Medicinal Chemistry-I Page 57

Experiment No:________ Date :

__________________________________OR_________________________
Aim: To perform assay of furosemide.
Reference: A Practical book of Medicinal Chemistry by Dr Abhishek Tiwari, Nirali Prakashan
anpp 66.
Apparatus and instrument: Volumetric flask, measuring cylinder, analytical balance, weight
box, beaker, burette and conical flask
Reagents and chemicals: Furosemide tablets, 0.1 M NaOH, dimethyl formamide, bromothymol
blue
Procedure
Weigh accurately about 0.5g furosemide dissolve in 40ml of dimethyl formamide. Titrate with
0.1 M sodium hydroxide using bromothymol blue as an indicator. Carry out a blank titration.

Each 1 ml 0.1 M sodium hydroxide equivalent to 0.03307 gm of furosemide

KBIPER Medicinal Chemistry-I Page 58

Experiment No:________ Date :

 Study Question:
1 What is Diuretic.

2 Which type of reaction occur in assay of furosemide.

3 What indicator use in assay of furosemide.

4 Give the molecular formula of furosemide.

5 Give the molecular weight of furosemide.

6 Give the use of furosemide.

7 furosemide comes under which class.

8 Give the structure of furosemide.

KBIPER Medicinal Chemistry-I Page 59

Experiment No:________ Date :

Experiment No: 15
Aim: To determine the partition coefficient for distribution of benzoic acid between octanol and
water.

Apparatus: Iodine flask, Volumetric flask, conical flask, measuring cylinder, analytical balance,
weight box, beaker, burette, pipette

Reagents and chemicals: Benzoic acid, octanol, NaOH and bromothymol blue.

Principle
When a solute is distributed between two solvents which are immiscible so as to form two separate
layers in contact with each other, the ratio of the equilibrium concentration of the solute in two
solvents(C1/C2) will be a constant at a given temperature. This constant is called partition
coefficient for that system.

Procedure
Take two stoppered 250 ml iodine flasks A and B. fill burette with a saturate solution of benzoic
acid in octanol and transfer 30 ml benzoic acid solution in to flask A and 25 ml benzoic acid in to
flask B. Add 5 ml of pure octanol in to flask B only to make volumes same. Then pour 150 ml of
water each in to two flasks. Put the stopper and shake the flask by a mechanical stirrer for 5
minutes. Then let the flask rest for 15 minutes so that the layers separate. While waiting, fill one
burette with 0.1 N NaOH solution. From this transfer 10 ml in to the volumetric flask and make
up to 100 ml to get 0.01 N NaOH solution. Fill the second burette with this solution. From iodine
flask A, pipette out 5 ml of organic layer in to a conical flask (organic layer should not be
contaminated by the aqueous layer; close the upper tip of the pipette with finger while inserting
the bottom tip to the very bottom of the organic layer and only then remove finger). And titrate
against 0.1 N NaOH solution by using bromothymol blue indicator. Pipette out 40 ml of aqueous
layer from the same bottle in to the conical flask, and titrate against 0.01 N NaOH solution. Repeat
two titration for iodine flask B also as a duplicate. Calculate partition coefficient for each flask.

KBIPER Medicinal Chemistry-I Page 60

Experiment No:________ Date :

Calculation

Flask Volumes of 0.1 N NaOH Volumes of 0.01 N NaOH Partition coefficient

(V1) (V2) K
A
B

Concentrations are proportional to volumes of NaOH used.

Since this is a ratio, volumes may be used directly.

Volumes of 0.01 N NaOH= benzoic acid in 40 ml octanol= V1* 10* 8 ml

K= V1* 10* 8/V2

RESULT:

The partition coefficient of benzoic acid between octanol and water=

(1)_____________

(2)_____________.

KBIPER Medicinal Chemistry-I Page 61

Experiment No:________ Date :

 Study Question:
1 What is Principle of partition coefficient for distribution of benzoic acid
between octanol and water.

2 Give the molecular formula of benzoic acid.

3 Give the molecular weight of benzoic acid.

4 Give the use of benzoic acid.

5 Give the structure of benzoic acid.

KBIPER Medicinal Chemistry-I Page 62

Experiment No:________ Date :

Experiment No: 16
Aim: To determine the partition coefficient for distribution of Salicylic acid between octanol and
water.

Apparatus: Iodine flask, Volumetric flask, conical flask, measuring cylinder, analytical balance,
weight box, beaker, burette, pipette

Reagents and chemicals: Salicylic acid, octanol, NaOH and crystal violet.

Principle
When a solute is distributed between two solvents which are immiscible so as to form two separate
layers in contact with each other, the ratio of the equilibrium concentration of the solute in two
solvents(C1/C2) will be a constant at a given temperature. This constant is called partition
coefficient for that system.

Procedure
Take two stoppered 250 ml iodine flasks A and B. fill burette with a saturate solution of Salicylic
acid in octanol and transfer 30 ml Salicylic acid solution in to flask A and 25 ml Salicylic acid in
to flask B. Add 5 ml of pure octanol in to flask B only to make volumes same. Then pour 150 ml
of water each in to two flasks. Put the stopper and shake the flask by a mechanical stirrer for 5
minutes. Then let the flask rest for 15 minutes so that the layers separate. While waiting, fill one
burette with 0.1 M perchloric acid solution. From this transfer 10 ml in to the volumetric flask and
make up to 100 ml to get 0.01 M perchloric acid solution. Fill the second burette with this solution.
From iodine flask A, pipette out 5 ml of organic layer in to a conical flask (organic layer should
not be contaminated by the aqueous layer; close the upper tip of the pipette with finger while
inserting the bottom tip to the very bottom of the organic layer and only then remove finger). And
titrate against 0.1 M perchloric acid solution by using crystal violet indicator. Pipette out 40 ml of
aqueous layer from the same bottle in to the conical flask, and titrate against 0.01 M perchloric
acid solution. Repeat two titration for iodine flask B also as a duplicate. Calculate partition
coefficient for each flask.

KBIPER Medicinal Chemistry-I Page 63

Experiment No:________ Date :

Calculation

Flask Volumes of 0.1 M perchloric Volumes of 0.01 M Partition coefficient

acid (V1) perchloric acid (V2) K
A
B

Concentrations are proportional to volumes of perchloric acid used.

Since this is a ratio, volumes may be used directly.

Volumes of 0.01 M perchloric acid = salicylic acid in 40 ml octanol= V1* 10* 8 ml

K= V1* 10* 8/V2

RESULT:

The partition coefficient of salicylic acid between octanol and water=

(1)_____________

(2)_____________.

KBIPER Medicinal Chemistry-I Page 64

Experiment No:________ Date :

 Study Question:
1 What is Principle of partition coefficient for distribution of Salicylic acid
between octanol and water.

2 Give the molecular formula of Salicylic acid

3 Give the molecular weight of Salicylic acid.

4 Give the use of Salicylic acid.

5 Salicylic acid comes under which class.

6 Give the structure of Salicylic acid.

KBIPER Medicinal Chemistry-I Page 65