Pharmaceutical organic chemistry

MARRI LAXMAN REDDY INSTITUTE OF PHARMACY

(Approved by AICTE & PCI, New Delhi and Affiliated to J.N.T. U, Hyderabad)

Dundigal (M) Medchal (Dist) Hyderabad- 500043

PHARMACEUTICAL ORGANIC CHEMISTRY- I

LAB MANUAL

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 About MLRIP
To be an educational Institute of par excellence and produce competent pharmacy professionals
to serve the community through research and the ever-increasing needs of Industry.

1. Imparting quality education and innovative research for various career opportunities.
2. Creating conducive academic environment to produce competent pharmacy professionals.
3. Indoctrination of students adorned with high human values and make them aware of their
responsibility as health care professionals.

PEO 1: To produce graduates with sound theoretical knowledge and technical skills required
for their career opportunities in various domains.
Program PEO 2: To incite the students towards research and to address the challenges with their innovative
Educational
contributions for the benefit of the mankind.
Objectives
PEO 3: To instill the essence of professionalism, ethical commitment to become a health
care professional with sound integrity and adherence to the core human values in the service
of the society.

PROGRAM OUTCOMES
1. Pharmacy Knowledge: Possess knowledge and comprehension of the core and basic knowledge associated
with the profession of pharmacy, including biomedical sciences; pharmaceutical sciences; behavioral, social,
and administrative pharmacy sciences; and manufacturing practices.
2. Planning Abilities: Demonstrate effective planning abilities including time management, resource
management, delegation skills and organizational skills. Develop and implement plans and organize work to
meet deadlines.
3. Problem analysis: Utilize the principles of scientific enquiry, thinking analytically, clearly and critically,
while solving problems and making decisions during daily practice. Find, analyze, evaluate and apply
information systematically and shall make defensible decisions.
4. Modern tool usage: Learn, select, and apply appropriate methods and procedures, resources, and modern
pharmacy-related computing tools with an understanding of the limitations.
5. Leadership skills: Understand and consider the human reaction to change, motivation issues, leadership and
team-building when planning changes required for fulfillment of practice, professional and societal
responsibilities. Assume participatory roles as responsible citizens or leadership roles when appropriate to
facilitate improvement in health and well-being.
6. Professional Identity: Understand, analyze and communicate the value of their professional roles in society
(e.g. health care professionals, promoters of health, educators, managers, employers, employees).
7. Pharmaceutical Ethics: Honour personal values and apply ethical principles in professional and social
contexts. Demonstrate behavior that recognizes cultural and personal variability in values, communication
and lifestyles. Use ethical frameworks; apply ethical principles while making decisions and take
responsibility for the outcomes associated with the decisions.
8. Communication: Communicate effectively with the pharmacy community and with society at large, such
as, being able to comprehend and write effective reports, make effective presentations and documentation,
and give and receive clear instructions.
9. The Pharmacist and society: Apply reasoning informed by the contextual knowledge to assess societal,
health, safety and legal issues and the consequent responsibilities relevant to the professional pharmacy
practice.
10. Environment and sustainability: Understand the impact of the professional pharmacy solutions in societal
and environmental contexts, and demonstrate the knowledge of, and need for sustainable development.
11. Life-long learning: Recognize the need for, and have the preparation and ability to engage in independent
and life-long learning in the broadest context of technological change. Self-assess and use feedback
effectively from others to identify learning needs and to satisfy these needs on an ongoing basis.
 Pharmaceutical organic chemistry

List of Experiments

I. Systematic qualitative analysis of unknown organic compounds like

1. Preliminary test: Color, odour, aliphatic/aromatic compounds, saturation and

unsaturation, etc.

2. Detection of elements like Nitrogen, Sulphur and Halogen by Lassaigne’s test

3. Solubility test

4. Functional group test like Phenols, Amides/ Urea, Carbohydrates, Amines, Carboxylic
acids, Aldehydes and Ketones, Alcohols, Esters, Aromatic and Halogenated
Hydrocarbons, Nitro compounds and Anilides.

5. Melting point/Boiling point of organic compounds

6. Identification of the unknown compound from the literature using melting point/ boiling
point.

7. Preparation of the derivatives and confirmation of the unknown compound by melting

point/ boiling point.

8. Minimum 5 unknown organic compounds to be analysed systematically.

2. Preparation of suitable solid derivatives from organic compounds

3. Construction of molecular models

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INTRODUCTION

Good Laboratory Practices:

1. Wear laboratory apron while working.
2. Use clean glassware.
3. Use strong acid and alkali carefully.
4. Do not displace the reagents from their respective places.
5. Do not interchange pipettes from one reagent to another without thorough
cleaning.
6. Carry out the reactions carefully without harming the neighboring student.
7. Use the gas whenever necessary. Close the gas tap when not required.
8. Prepare your own reagents when necessary for correct results.
9. Do not consult your friends for doubts. Consults books and teachers for your
problems.
10. Take signature of your teachers for all your assignments.
11. Leave the laboratory well prepared for the experiment concerned.
12. Write the laboratory records regularly

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APPARATUS AND GLASS WARES

Before starting an experiment cleaning of glassware, equipment and bench is necessary.

The bench should be kept clean and remove any solid or liquid chemicals. Glass ware are first
washed with chemicals and then rinsed with distilled water before use. The outer surface may be
dried with a cloth or filter paper. All the apparatus used in particular experiment should be kept
together on the bench to avoid confusion in determining the duplicate experiments. Excess of
apparatus which are not in use should be removed from the bench. All the solutions, precipitates
and filtrates should be labeled to avoid confusion and covered to prevent contamination of the
contents. Reagent bottles must be returned on the reagent shelves immediately after use.
Experiment observations must be recorded in a shift covered notebook containing index page
and remaining one sided line pages. The record must be concluded with calculations and results.
Glassware:
The convenient unit to measure large volumes of liquid is “liter”. The liter and milliliter
are sufficiently precise for the requirement of titrimetric analysis. The capacity of a glass vessel
may vary with temperature. Therefore, the temperature of the experiment is noted.
Graduated apparatus:
Graduated flasks, burettes and pipettes are the most commonly used apparatus in
volumetric analysis. All these glassware must be perfectly clean from grease to get exact results.
Many detergents are available to clean glass apparatus. Saturated solution of powdered sodium
and potassium dichromate in conc. sulphuric acid is also used for cleaning purpose.
Volumetric flask:
A volumetric flask is a flat-bottomed, pear-shaped apparatus with a long-narrowed neck.
A thin line mark around the neck indicates the volume that it holds at a certain definite
temperature,
Usually, 20 degrees the capacity and the temperature are clearly marked on the
flask. Both the front and the back of the mark should be seen as a single line to avoid the errors
as parallax, when making the final adjustments. The lower edge of the meniscus should be
tangential to the graduation mark. A small change in the volume is easily detected in a long
narrow neck. Volumetric flasks are available in capacities of 1, 2,5,10,25,50,100,500,1000,2000
and 5000 cm3.

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Pipette: Pipettes are of two types:

1. Transfer/bulb/volumetric pipette: which have one mark and withdraw a small
and constant volume of solution.
2. Measuring/graduated pipettes: which are graduated and used to deliver various
small volumes.
Filtration Apparatus:
• A conical flask and funnel fitted with a filter paper is usually used for filtration. The
funnel should have angle nearby 60 degrees and a long stem.
• Sintered glass crucibles are made up of resistance glass and have porous disk fused into
the body of crucible. The filtered disc is made up of various pore diameters as indicated by the
numbers from 0-5.
• Buchner funnel is used to filter large quantities of materials. A good quality of filter
paper is placed on a paper with sintered glass.

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Narrow neck RBF Long neckRB flask Pear shaped RB flask Volumetric Flask

Round Bottomed Flask Round Bottomed Flask Two neck R.B Flask Three neck R.B Flask

Erlenmeyer flask Buchners Flask Beaker Measuring Cylinder

Thermometer adapter Plain bend Distillation adapter Vacuum adapter

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Pear shaped globe shaped burette

seperating funnel seperating funnel titration burette stand with clamp Heating Mantle

clavengers apparatus Soxhlet with claisen adapter

Liebig condenser Allihn condenser set
Condenser condenser

Cone powder funnel

Short Stem Funnel
Buchners Funnel
Long Stem Funnel

Thermometer

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DETERMINATION OF MELTING POINT

AIM: To determine the melting point of given liquids.

APPARATUS: Thermometer, Capillary tube. Melting Point Apparatus.
PRINCIPLE: Melting point is the change from the highly order or arrangement of particles in
the crystalline lattice to the more random arrangement that characterize a liquid. Melting occurs
when the temperature reaches at which the thermal energy of the particles is great enough to
overcome the intra crystalline forces that holds them in position.
PROCEDURE: A capillary tube is taken and its one end is closed by introducing into the tip of
flame. Capillary tube is kept rotating during heating for the closing to be uniform. Care is taken
to prevent the formation of bulb during heating.

METHOD –I: The substance whose melting point is to be determined is introduced into
capillary tube so that it occupies at least 3-5 mm of the length. The substance and capillary tube
must be dried before actual procedure begins.
The filled capillary tube is tied to a thermometer with a thread such that the substance is
against the mercury bulb of thermometer. This is introduced into a melting point apparatus which
is filled with liquid paraffin or concentrated sulphuric acid. The liquid is heated gently using a
blue flame. Care is taken, so that the temperature rise uniform is not more than 1-20C per minute.

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It is kept on heating until the substance in capillary tube begins to melt. Flame is removed at this
stage and temperature is observed at which melting had begun and where it had melted.
METHOD II: Introduce small quantity of sample in sealed capillary tube and keep it in a
melting point determining apparatus along with thermometer through hole of apparatus and note
down the temperature where the sample starts melting and then finally where it completely
melts, this gives melting point range.

REPORT: Melting point range of various compounds is

Naphthalene : 80-82 0C
- napthol : 94-96 0C
- napthol : 120-123 0C
Benzoic acid : 120-1210C
Cinnamic acid : 133 0C
Anthracene : 210 0C
Benzophenone : 47-48 0C
Benzamide : 117-119 0C

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DETERMINATION OF BOILING POINT

AIM: To determine the boiling point of given liquids.

APPARATUS: Boiling tube, Thermometer, Capillary tube.

Principle: The boiling point of a substance is the temperature at which the vapor pressure of
the liquid equals the pressure surrounding the liquid and the liquid changes into a vapor. The
boiling point of a liquid varies depending upon the surrounding environmental pressure.
Procedure: This method is Siwoloboffs method. When reasonable amounts of liquid compound
are available, the boiling point is readily determined by slowly distilling the material from a
suitable flask and recoding the temperature at which the bulk of compound is distilled. For
smaller quantities of the liquid, micro methods may be used.
Siwoloboffs Method:
In this procedure two tubes are used. One is the ordinary capillary tube i.e., 90
mm – 100 mm long and 1 mm in diameter and the other one is 100 mm long and 4-5 mm in
diameter. Both the tubes are closed at one end. A small quantity of a liquid (0.5ml) is placed in a
wide tube and the capillary tube with sealed end is introduced into the liquid. This tube is then
attached to the thermometer with a thread and the thermometer is immersed in the bath of
melting point apparatus. Heat the tube gradually. As the heating is continued, there will be a
slow escape of bubbles from the end of capillary tube at a low speed in the beginning due to
thermal expansion of the trapped air. Continue heating slowly. When the temperature of the
liquid in the ignition tube equals to its boiling points, a rapid and continuous escape of the
bubbles will occur. The reading of the thermometer, when rapid and continuous streams of
bubbles first emerge from the capillary tube, is the boiling point of the liquid.

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A more accurate result is obtained by removing source of heat, when the rapid
stream of bubbles rises from the end of the capillary tube. Note the temperature at which the
bubbles just fail to come out of the capillary and the liquid stats to enter the capillary. This
temperature is taken as boiling point of the liquid.

Carbon tetrachloride - Literature Value - 77 0C

Chlorobenzene - Literature Value - 132 0C
Acetone - Literature Value - 56 0C

REPORT: Boiling point range of various compounds was found to be as

Acetone : 56
Glycerine : 290
Ethanol : 78
Acetic anhydride : 140
Benzene : 80
Benzyl alcohol : 206

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PREPARATION OF p–BROMOACETANILIDE FROM ACETANILIDE

Aim: To synthesize p –Bromoacetanilide from Acetanilide

CHEMICALS APPARATUS
Acetanilide 4 gm Conical Flask,
Bromine – 1.5 ml, Glass rod
Glacial acetic acid – 15 ml Beaker,
Sodium bisulphate Buckner funnel
Sodium metabisulphite Burette.

Principle:

IUPAC Name: N-(4-bromophenyl) acetamide

Synonyms: 4-bromoacetanilide, mono bromoacetanilide, bromoanilide
Chemical Formula: C8H8BrNO
Molecular weight: 214.06 g/mol

Structure
CH 3

HN O

Br
p-bromo acetanilide
Explanation:
Acetanilide on reaction with bromine in presence of Glacial acetic acid (Nuclear
bromination) to give p-bromoacetanilide. This mechanism is a classic example of Electrophilic
aromatic substitution. An amine may lead to di- and tri- substituted products. If an amide is
used in place of the amine, mono substitution usually predominates (the electron-withdrawing
carbonyl group makes the benzene ring less nucleophillic). This Ortho-, Para- directing group
will tend to only add groups para- to itself because of the steric bulk of the amide group.

Reaction
O
O
Br2 HN CH3
HN CH 3
Glacial. CH 3COOH

acetanilide Br
p- bromoacetanilide

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Procedure:
Dissolve 4 gm of acetanilide in glacial acetic acid (15ml) in a clean conical flask and add
slowly drop wise a solution of bromine (1.5ml) in acetic acid (5ml). Shake the flask vigorously
and cool during the addition. The addition of bromine solution allows the reaction mixture to
stand for 30 minutes with frequent shaking. Pour the contents of the conical flask in to beaker
containing cold water (200ml) with constant stirring and rinse the conical flask and transfer the
contents into same beaker. If the product separates, it is colored, and then add sodium sulphite or
sodium metabisulphite until it becomes colorless. Then filter through the Buchner funnel. Wash
with cold water and dry.
Recrystallization:
A pure sample of p-bromo-acetanilide can be obtained as colorless crystalline solid on
recrystallization from hot alcohol.

Percentage yield:

S. No Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

Description:

Melting point : 168 0C,

Solubility : insoluble in cold water,
: Sparingly soluble in hot water,
: Soluble in benzene, chloroform, ethyl acetate,
: Moderately soluble in alcohol.
Uses: 1. Used as an Analgesic agent
2. Used as Anti-pyretic agent.

Precaution: Handle bromine carefully as it digests the bones and produces irritation to the eyes

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Result: P –Bromoacetanilide was synthesized from Acetanilide, the % yield was found to
be ________ % w/v and the melting point was found to be ________.

Viva questions:
1) What is the mechanism involved in the preparation of parabromo acetanilide?
2) What is the use of sodium sulphite or sodium metabisulphite?
3) Which of the reagents should be taken excess and why?
4) What are the uses of P-bromoacetanilide?
5) What is the principle involved it?

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PREPARATION OF PARA-BROMOANILINE FROM PARA- BROMOACETANILIDE

AIM: To prepare p-Bromoaniline from p-Bromoacetanilide by Hydrolysis process.

CHEMICALS APPARATUS
p-Bromoacetanilide- 18 gm Round bottom flask
Alcohol – 35 ml Condenser
5% NaOH Adaptor
Con. H2SO4 – 1 to 2 drops Conical Flask, Funnel

Principle:

IUPAC name: 4-bromoaniline

Synonyms: p-bromoaniline, 4-bromobenzenamine, p-bromophenylamine
Chemical Formula: C6H6BrN
Molecular weight: 172.02 g/mol
Structure:
NH 2

Br
p-bromo aniline
Explanation:
p-Bromoaniline cannot be prepared by bromination of aniline, since the major product of
bromination is 2,4,6-tribromoaniline. Therefore, the amino group is protected by mineral acids
and then bromination gives p-Bromoacetanilide as the major product which on hydrolysis gives
p-Bromoaniline in the presence of mineral acids.
Reaction:
O
NH 2
HN CH 3
NaOH, HCl O

H2 O + H3 C OH
Br
Br
p- bromoacetanilide p-bromo aniline acetic acid
Procedure:
Dissolve 18 g (0.084 mol) of p-Bromoacetanilide in 35 ml of boiling ethanol contained in
a 250 ml round bottomed flask equipped with a reflux condenser. With the aid of a pressure
equalizing dropping funnel add 22 ml of concentrated hydrochloric acid down the condenser in
small portions to the boiling solution, Reflux for 30-40 minutes or until a test portion remains
clear when diluted with 150 ml of water, and fit the flask with a condenser set for downward

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distillation. Distil the mixture from an air bath and collect about 100 ml of distillate the latter
consists of ethyl acetate, ethanol and water. Pour the residual solution of p-bromoaniline
hydrochloride in to 100 ml of ice-water, and add with vigorous stirring, 5% sodium hydroxide
solution until just alkaline. The p-bromoaniline separates as on oil, which soon crystallizes. Filter
the crystals and wash with cold water and dry in the air upon pads of filter paper.

Recrystallisation: A pure sample of p-bromo-aniline can be obtained as colourless crystalline

solid on Recrystallisation from hot alcohol.

Percentage yield:

S. No Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

Description:
Melting point : 60 to 64 0C
Solubility : soluble in water

Uses: 1. Used as an Analgesic agent

2. Used as Anti-pyretic agent.

Result: P –Bromoaniline was synthesized from hydrolysis of p-Bromoacetanilide, the

percentage yield was found to be ________ % w/v and the melting point was found to be
________.

Viva Questions:

1. Which compound is produced on hydrolysis of p-bromo acetanilide?

2. Define bromination.
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3. Which compound is produced on bromination of aniline?

4. Give the IUPAC name of aniline.
5. What is the pharmaceutical significance of p-Bromoaniline?

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PREPARATION OF m- DINITROBENZENE

AIM: To synthesize m- Dinitrobenzene from Nitrobenzene.

CHEMICALS APPARATUS
Conc. Nitric acid – 7 ml, Measuring jar,
Conc. H2SO4 – 8 ml Round bottom flask
Nitro Benzene – 6 ml Condenser
Distilled Water – 100 ml Beaker, Funnel etc.

Principle:

IUPAC Name: 1, 3- dinitrobenzene

Synonyms: Dwunitrobenzene; Binitrobenzene; o-Dinitrobezene; m-dinitro-benzene
Chemical Formula: C6H4N2O4
Molecular weight: 168.02 g/mol
Structure:
O O-
N+

O
N+
O-
m-Dinitrobenzene
Explanation:
Nitrobenzene on nitration with Conc. Nitric acid in the presence of sulphuric acid 60 0C
gives m-dinitrobenzene. The nitro group present on the benzene ring is electron deficient or
electron withdrawing or ring deactivating group, due to the presence of this group the electron
density on Ortho and Para positions becomes very less when compare to that of the meta
position. So, its attacking position may be Meta. Hence, Nitrobenzene on nitration will result in
the formation of m-dinitrobenzene.

Reaction:
NO 2 NO2
Conc. H 2SO 4

Conc.HNO3
NO2
Nitro benzene m- dinitrobenzene

Procedure:

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Place 7 ml of Nitric acid in 100 ml of round bottom flask and add 8 ml of Conc. H2SO4 in
small portion with shaking, Keep the reaction mixture cool while adding by immersing the flask
in crushed ice or ice water. Introduce 6 ml of nitrobenzene in small portions, cooling the flask in
ice or ice water, shake the flask to ensure thorough mixing after each addition of nitrobenzene,
do not allow the temperature of the mixture to rise above 55 0C.
When all the nitrobenzene has been added, fit a reflux condenser to the flask. Heat on
water- bath and maintain at 60 0C for 30-40 minutes. Remove the flask from water bath for time
to time and shake it vigorously to ensure good mixing to the immiscible liquids layers. Later
remove the flask from water bath and pour the contents of the flask into 100 ml of cold water
taken in a beaker. Stir the mixture well to remove acids as much as possible from the m-
dinitrobenzene and allow standing. When the m-dinitrobenzene settles to the bottom, pour off the
acid layer as completely as possible and transfer the remaining liquid to separating funnel.
Again, wash with 50 ml of water and allow it to settle. Collect the lower layer of m-
dinitrobenzene into a dry test tube or boiling tube adds anhydrous calcium chloride which acts as
dehydrating agent. After 24 hours a clear liquid with pale yellow color crystals will be obtained.
Collect the crystals by filtration.

Recrystallization:

A pure sample of m-dinitrobenzene can be obtained as pale-yellow color crystalline solid

on Recrystallization from alcohol.

Precaution: m-dinitrobenzene is toxic and do not inhale its vapor. Let it not fall on your skin.

Percentage yield:

S.no Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

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Description:
Melting Point : 89.6 0C
Solubility : freely soluble in chloroform, ethyl acetate

Uses: 1. Used in pesticides.

2. Used in dyes.

Result: m-dinitrobenzene was synthesized from nitrobenzene, the percentage yield was found to
be ________ % w/v and the melting point was found to be ________.

Viva questions:
1) What is nitration mixture?
2) What is the name of the reaction?
3) What is the function of sulphuric acid?
4) What is the mechanism of reaction?
5) What is the function of calcium chloride?
6) What is electrophile in this experiment?
7) What precautions are taken in handling nitrobenzene?

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PREPARATION OF N-BUTYL ACETATE FROM N-BUTYL ALCOHOL

AIM: To prepare and submit the pure sample of n-butyl acetate.

CHEMICALS APPARATUS
Glacial acetic acid – 15ml Round bottom flask
N-butyl alcohol – 12ml Reflux condenser
H2SO4 – 1 ml Beaker
5% Na2Co3 solution, Water bath, Glass rod,
Anhydrous sodium sulphate Boiling tube, Measuring jar,
Separating funnel.

Principle:

IUPAC Name: n - Butyl acetate

Synonym: n-butyl ethanoate, Acetic acid-butyl ester,
Chemical Formula: C6H12O2
Molecular Weight: 116.16 g/mol
Structure

H3 C O CH 3
n-butyl acetate

Explanation:
n-butyl alcohol undergoes esterification with Glacial acetic acid in the presence of
sulphuric acid to give n-butyl acetate.

Reaction

Conc. H2 SO4 O
HO CH 3 + + CH3COOH
H3 C O CH 3
n-butanol
n-butyl acetate

Procedure:

Take 12 ml of n-butyl alcohol into a clean, dry round bottom flask and 15 ml of glacial
acetic acid and followed by 1 ml of concentrated sulphuric acid. Fit the round bottom flask with
reflux condenser and heat it on non luminous flame for about 1 hour. Cool the reaction mixture
and pour it into cold water with stirring. A pleasant smelling, light colorless oil separates on the
surface of water as upper layer. Transfer into separating funnel and add dilute sodium carbonate

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solution. Finally wash it with cold water and draw into a dry test tube. Dry over anhydrous
sodium sulphate. Further purification can be done by distillation method.

Percentage yield:

S.no Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

Description:
1. Boiling Point: 126 0C
2. Solubility : miscible in ethanol, soluble in acetone, CHCl3

Uses: 1. Used in foods as anti flavouring agent.

2. used in the manufacture of lacquers, artificial leather, photographic films, plastics, and
safety glass.

Result: n- butyl acetate was synthesized from n- butanol, the percentage yield was found to be
________ % w/v and the boiling point was found to be ________.

Viva questions:

1) How is the concentrated n-butyl alcohol separated?

2) Why should we use sodium carbonate?

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3) Why do we use anhydrous sodium sulphate?

4) Write the principle involved in preparation of N-butyl acetate.
5) Write the structure of N-butyl acetate.

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PREPARATION OF β- NAPTHYL METHYL ETHER (NEROLIN) FROM β- NAPTHOL

AIM: To synthesize and submit the Nerolin from β- napthol

CHEMICALS APPARATUS
-Naphthol – 3.6 gm Conical flask
Dimethyl sulphate – 2.5 ml Glass rod
10% Sodium hydroxide – 20 to 30 ml Water bath,
Con. H2SO4 – 1 to 2 drops Measuring cylinder, Beaker.

Principle:

IUPAC name : 2-Methoxynaphthalene

Synonyms: 2-Naphthol methyl ether, 2-Naphthyl methyl ether, β-Methoxynaphthalene,
β-Methyl naphthyl ether, β-Naphthol methyl ether;
Molecular weight: 158.09 g/mol
Structure:

O
CH 3

2-methoxynaphthalene
Explanation:
β- Naphthol undergoes methylation in the presence of dimethyl suphate and NaOH to
give β- napthyl methyl ether which is also called as nerolin. The mechanism involved in this
reaction is nucleophillic substitution reaction.

Reaction:

OH NaOH OCH3
+ (CH 3) 2SO4

beta naphthol Nerolin

Procedure:

Take 3.6 gm of -Naphthol in a conical flask and dissolve it in 20-30ml of 10% Sodium
hydroxide solution. Add 2.5 ml dimethyl sulphate while the mixture is cooled on ice. Warm the
mixture for 10-15 minutes at 60-70 0C and allow it for cooling. Filter off the separated nerolin at
the vacuum pump, wash it with 10% sodium hydroxide solution and thoroughly with cold water.
Dry the product and recrystallize it from either rectified spirit or methylated spirit.

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Note:
Dimethyl sulphate decomposes to give sulphuric acid. Therefore, the reagent must be free
from sulphuric acid. Thus, purification of dimethyl sulphate is required, which is done by
shaking with 10% sodium carbonate or sodium bicarbonate.
(These are preferred to sodium hydroxide because on neutralization of acid, they release
CO2, which is visible to our eyes).

Caution: Dimethyl sulphate is highly corrosive and irritating substance. Therefore, contact to skin
is to be avoided.

Percentage yield:

S.no Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

Description:
1. Melting Point : 70- 72 0C
2. Solubility : 1. it is soluble in alcohol
2. Insoluble in water and dipropylene glycol.

Uses: 1. Used in paints and varnishes.

2. used as a stabilizer found in gunpowder.

Result: β-Naphthol methyl ether was synthesized from β-Naphthol, the percentage yield was
found to be ________ % w/v and the boiling point was found to be _______.

Viva questions:

1) What is the role of sodium hydroxide in this reaction?

2) Why should we purify dimethyl sulphate before use and how it is done?

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3) What is the advantage of washing dimethyl sulphate with sodium carbonate or sodium
bicarbonate rather than with NaOH?
4) The precipitate of nerolin is washed with NaOH solution, why?
5) Why should we wash the precipitate of nerolin thoroughly with water?
6) What is the methylated spirit?
7) What is the test for purity of -naphthol methyl ether or nerolin?
8) What is the function of dimethyl sulphate?

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PREPARATION OF OF IODOFORM FROM ETHYL ALCOHOL

AIM: To synthesize and submit iodoform from Ethyl alcohol.

CHEMICALS APPARATUS
Alcohol – 30 ml Conical flask,
10% NaOH solution – 30 ml Measuring cylinder
Iodine solution – q.s Glass rod, Thermometer, Funnel

Principle:

IUPAC Name: Triiodomehane

Synonyms: Iodoform
Chemical Formula: CHI3
Molecular Weight: 393.73 g/mol
Structure:
I
I
I

IODOFORM

Explanation:

Many substances containing either acetyl or ketone group on treating with KI and NaOH
yield Iodoform and its formation is used as test for identifying these groups. Examples for acetyl
groups are ethanol, isopropyl alcohol etc. The compounds which contain ketone group are
acetone, Pyruvic acid etc. It is synthesized by Haloform Reaction, where a Haloform (CHX3,
where X is a halogen) is produced by the exhaustive Halogenation of any of the following 4
groups.
• Methyl ketone: CH3COR,
• Acetaldehyde (CH3CHO),
• Ethanol (CH3CH2OH)
• secondary alcohols (CH3CHROH, where R is an alkyl or aryl group).
Any one of these four kinds of organic compounds gives Haloform reaction with iodine
and sodium hydroxide.

Reaction:

(O) I2/KI
CH 3OH CH 3CHO CI3CHO + NaOH CHI3 + HCOONa
Methanol Iodoform

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Procedure:

Place 30 ml of alcohol in a conical flask and add 30 ml of 10% NaOH solution to it. Add
iodine solution in small quantities while shaking until a yellow colour persists, which is due to an
excess of iodine. Warm the contents of the flask on water bath and maintain the temperature
between 50 -60 0C. After 10-15 minutes of heating, remove the conical flask from the water bath,
cool and collect iodoform by filtration.

Recrystallization:

Recrystallize it either from ethyl alcohol or methyl alcohol. Take prepared iodoform into
a boiling tube and add sufficient amount of methanol. Allow to cool to room temperature for 15
to 20 minutes until crystals of Iodoform settle at the bottom. The product should not be dissolved
in methanol. Filter and collect crystals of iodoform.

Note:
While heating the iodoform and methanol in a water-bath, small porcelain pieces are
added to the boiling tube, so as to prevent the bumping of the liquid.

Iodine Solution: Dissolve 2 gm of Iodine and 3 gm of KI in water to produce 100ml.

NOTE: Iodine solution is a solution of Iodine in potassium Iodide solution (KI). Iodine is
insoluble in water. It dissolves in a solution of potassium Iodide (KI) due to formation of KI3.

KI+I2→ KI3
Percentage yield:

S.no Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

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Description:
Melting Point : 119-121 0C
Solubility : soluble in ether, CHCl3, alcohol, slightly soluble in H2O

Uses: 1. Used as an antiseptic.

2. Used as a disinfectant.

Result: Iodoform was synthesized from Iodine and ethyl alcohol, the percentage yield was found
to be ________ % w/v and the melting point was found to be ________.

VIVA QUESTIONS:

1. Why should you use an excess of iodine in this experiment?

2. How do you know that you have used an excess of iodine?
3. Give examples for compounds that show positive result for iodoform test.
4. Give examples for compounds that do not show positive result for iodoform test.
5. What is Haloform reaction? Give one example.

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PREPARATION OF 2, 4, 6- TRIBROMOANILINE FROM ANILINE

AIM: To prepare 2,4,6- tribromoaniline from aniline by Bromination process.

CHEMICALS APPARATUS
Aniline Round bottom flask
Glacial Acetic acid – 35 ml Reflux Condenser
Bromine in Glacial Acetic acid Adaptor, Measuring Cylinder
Ethanol, Conical Flask, Funnel

Principle:

IUPAC name: 2, 4, 6- tribromoaniline

Synonyms: 2, 4, 6-Tribromobenzenamine, Tribromoaniline
Chemical Formula: C6H4Br3N
Molecular weight: 329.81 g/mol
Structure:
NH2
Br Br

Br
2,4,6- tribromoaniline

Explanation:

Aniline undergoes Bromination with bromine in the presence of glacial acetic acid and
results in the formation of 2, 4, 6- tribromoaniline, at first Bromine attack the ring between the
ortho carbon and the carbon bearing the amine group. The electron pair of the amine moves to
the C-N bond (making a double bond and a quadravalent ammonium group), the electrons of the
ring move to one of the bromine atoms of Br2, causing Br - to form. The ortho carbon now bears
a hydrogen atom and a bromine atom; and the positive charge is delocalized via resonance
around the ring and the ammonium to make a stable cation. Once the Br - plucks the hydrogen
from the ring, bromoanilide and HBr are formed. Since the amino group is an activating o-p
director, this will happen twice more to give 2, 4, 6-tribromoaniline as final product provided
sufficient Br2 is present.

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Reaction:

NH2
NH 2
Glacial CH 3COOH Br Br
+ 3 Br 2

Aniline Br
Bromine
2,4,6- tribromoaniline

Procedure:

Take 2.5 ml of aniline, 10 ml of glacial acetic acid in a 250 ml round bottomed flask. 4.2
ml of bromine is added drop wise (which is previously dissolved in 10 ml of glacial acetic acid)
with constant stirring while keeping the round bottomed flask in ice bath. A solid mass is
obtained which is filtered and washed with cold water. Recrystallize the product from dilute
alcohol to obtain colorless crystals of 2, 4, 6-tribromoaniline.

Recrystallization: A pure sample of 2, 4, 6-tribromoaniline can be obtained as colorless

crystalline solid on Recrystallization with small amount of activated charcoal from ethanol.

Percentage yield:

S. No Chemical name Quantity required Quantity taken

Percentage yield

Theoretical yield =

Description:
Melting point: 120-122 0C
Solubility : slightly soluble in water, freely soluble in ether and ethyl acetate

Uses: 1. Used in the preparation of dyes, polyamides,

2. Used in the preparation of antioxidants.

Result: 2, 4, 6-tribromoaniline was synthesized from aniline, the percentage yield was found to
be ________ % w/v and the melting point was found to be ________

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ANALYSIS OF ORGANIC COMPOUNDS

IDENTIFICATION OF ORGANIC COMPOUNDS

EXPERIMENT OBSERVATION INFERENCE

PRELIMINARY TEST
Solid The given compound may be carboxylic
1. Physical state acid
Liquid The given compound may be alcohol,
ether, ester, aliphatic amine aldehydes,
ketone, hydrocarbon
Colorless The compound may be carbohydrates,
2. Colour aldehydes, ketones, carboxylic acids,
esters, alcohols or hydrocarbons

Pale yellow The compound may be nitro compounds,

quinines(or) iodoform
Pink Alpha-Naphthol
Beta- Napthol (or) Resorcinol
Dark brown(or) Reddish Phenols or amines
brown
Yellowish orange Nitro aniline
Pleasant odour It may be alcohol, aromatic hydro carbons
3. Odour or alkyl halides
Fruity odour Esters
Phenolic odour Phenols
Deep sweet smell Chloroform
Odour of bitter almond Nitrobenzene Benzaldehyde or nitrotoulene
Irritating odour Aliphatic acids, acid chlorides or
formaldehyde
Fishy odour Amines
4. Litmus test: Colour of the blue litmus is Acidic compound
A small quantity of the changed to red
substance is shaken well Colour of the red litmus Basic compound
with 5ml of water and changes to blue
solution and tested with No change in the colour of Neutral compound
litmus paper litmus paper

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Ignition test Smoky flame Aromatic compounds or

Heating on wire or spatula compounds containing more than
six carbon atoms
No smoky flame Aliphatic compounds
Greenish flame Compounds containing more
number of halogen atoms
Charring and smell of burnt Carbohydrates or citric acid.
sugar
Test for unsaturation The yellow color of the Unsaturated compound
1. A little substance is solution is decolorized
dissolved in 1ml of water Yellow color of the solution Saturated compound
&treated with 3 to 4 drops of remains
bromine water Yellow color of the sol is Unsaturated compound like
decolorized and precipitate amine or phenol.
formation is seen
2. A little substance is Pink color of the sol is Unsaturated compound
dissolved in 1ml of water decolorized
&treated with 3-4 drops of Pink color of the solution Saturated compound.
1% KMnO4 remains

Test for aliphatic or aromatic Substance burns with smoky Presence of aromatic compound
compound flame
1. Flame test Substance burns with non Presence of aliphatic compounds
luminous flame
Substance burns with Carbohydrates
charring
2. Nitration test: A small amount Yellow ppt or solution Presence of aromatic compound
of the substance is heated with a No yellow colour is seen Presence aliphatic compound.
mixture of 1ml of conc. H2SO4
&1ml of con HNO3

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SOLUBILITY CHART

REAGENT AND TEST CLASS GROUP OF COMPOUNDS

Soluble in cold or hot Neutral, acidic or basic. Lower members of series.
water. (If the unknown is (Test with litmus or Neutral, e.g., alcohols; Acidic, e.g., acids,
soluble do NOT perform universal indicator phenols; Basic, e.g., amines
solubility tests below) paper)
Soluble in dil. HCl Most amines (except III amines
Basic
with only aromatic groups)
Soluble in dil. NaOH Acidic Most acids, most phenols.

Soluble in NaHCO3 Strongly acidic Most carboxylic acids.

Hydrocarbons, nitrohydro-
Insoluble in water, acid carbons, alkyl or aryl halides, esters and
Neutral
and alkali ethers. Higher molecular weight alcohols,
aldehydes and ketones

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DETECTION OF EXTRA ELEMENTS

Lassaignes Test:

Procedure for preparation of Sodium Fusion extract:

Take in a sodium fusion tube, small pieces of shining freshly cut metallic sodium in
excess which is previously dried between the folds of filter paper. Heat the tube on the flame so
that sodium metal melts. Remove it from the flame and allow it to stand, so that sodium occupies
the bottom portion of the tube. Add a pinch of the substance to be tested and heat the tube once
again, gently with removing the tube from the flame time to time Then heat the tube strongly
until it becomes red hot and drops it immediately in about 10-15 ml of distilled water, taken in a
clean mortar. Close the mortar with wire gauge. After the tube has broken and the reaction
stopped. Remove the wire gauge and triturate with a pestle. Filter the contents and the filtrate is
called sodium fusion extract.

TEST OBSERVATION INFERENCE

Test for Nitrogen:
Take 2-3 ml of sodium fusion extract in a If Prussian blue Indicates presence
test tube Add 0.1-0.2 gm of ferrous sulphate color appears of nitrogen.
crystals. Heat the mixture gently with shaking until
it boils without cooling, add sufficient amount of
dilute sulphuric acid to dissolve the Iron hydroxides
and give the solution of acidic medium.

Test for sulphur:

Acidify 2ml of sodium fusion extract with If black precipitate Indicate the
acetic acid and add a few drops of lead acetate of lead sulphide is presence of
solution. formed sulphur.
Test for halides:
Acidify a portion (1 mL) of the 'fusion'
filtrate with 2N nitric acid, and if nitrogen and/or (a) If the carbon Indicates presence
sulphur are present, boil for 1 - 2 minutes. * Cool tetrachloride layer of chlorine.
and add aqueous silver nitrate (1 mL), compare with remains colourless.
a blank. Formation of a heavy, white or yellow
precipitate of silver halide indicates halogen. If a (b) If the carbon Indicates presence
positive result is obtained: acidify the remaining tetrachloride layer is of bromine
portion of the 'fusion' filtrate with dilute sulphuric brown.

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acid, boil and cool. Add carbon tetrachloride (1 mL)

and a few drops of freshly prepared chlorine water. (c) If the carbon Indicates presence
Shake the mixture. tetrachloride layer is of iodine.
violet.

NOTE: *If nitrogen and/or sulphur are also present, the addition of silver nitrate to the acidified
'fusion' solution will precipitate silver cyanide and/or silver sulphide in addition to the silver
halides. The removal of hydrogen cyanide and/or hydrogen sulphide is affected by boiling the
'fusion' solution.

PRECAUTIONS TO BE TAKEN DURING PREPARATION OF SODIUM FUSION

EXTRACT

1) Sodium is always stored under kerosene.

2) Sodium should not be exposed to skin because it reacts with water and produces a
burning sensation.
3) The mortar should be covered with a wire gauge after plunging the red hot tube into it; to
avoid fire caused by excess of unreacted sodium on reaction with water.
4) The metal absorbs small quantities of moisture, the outer layer is encrusted with sodium
hydroxide therefore while taking the metal for this test, one must scrap off the outer
surface with a blade or knife, until sodium hydroxide is removed and a bright luster of
sodium metal persists.
5) Always handle the metal with a forceps.
6) While heating one must be careful the metal does not come out of the tube and fall on the
skin or eyes.

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FUNTIONAL GROUP TEST

ANALYSIS OF PHENOLS

Experiments Observation Inference

Sodium carbonate solution Test: Add 0.1 gms of No effervescence and no gas May be phenol
substance to about 5ml of Na2CO3 solution taken in a evolved and it changes the
test tube. A drop of test sample is placed on blue litmus blue litmus to red.
paper.
May be phenol,
Ferric Chloride Test: Take a very dilute solution of Blue or violet, green-violet resorcinol,
phenol; add a drop of ferric chloride solution. color catechol,
p-bromophenol

Phthalein Test (or) reaction with phthalic Indicates presence

Anhydride: Place in a dry test tube about 0.2gms of Colourless solution turns of phenolic
phenol, add equal quantity phthalic anhydride, add 3 blue or green in NaOH compound. (Except
drops of sulphuric acid and heat for about 10 minutes medium. p-cresol)
until red homogenous mass is formed. pour the mixture
into water taken in a beaker and add NaOH solution in
excess.

Action of Bromine water / Bromine water test: In a Reddish brown colour is Presence of
concentrated substance of phenol add Bromine water discharged and a white or phenolic
gradually. pale yellow ppt is formed compound.

Liebermann Reaction: Minute crystals of sodium Blue-green color is Indicates presence

nitrite are taken in a clean test tube, add 0.1 gms of developed on dilute the of phenol with free
phenol and heat gently for 20 seconds Allow it to cool contents with H2O the color para position
and add 1ml of Conc. H2SO4.The test tube is rotated to changes to red which turns
mix the contents to blue with NaOH.

To a small amount of substance add 2ml of NaOH and

add 2 drops of in a test tube and add 2ml of aqueous Orange dark red color
NaOH and 1 drop of chloroform and warm it. observed. Presence of phenol

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Azo-Dye formation: Dissolve 2-3 drops of aniline in Brown red colour is Presence of 1-
1ml of conc. HCl and 3 ml of water. Shake it to observed. napthol
dissolve any HCl cool in ice, add a few drops of
sodium nitrite solution. Add a thick cool diazonium
solution to phenol in excess of NaOH solution.

ANALYSIS OF AMIDES

1) Action of NaOH solution: To a May be amide

little substance, add a little Smell of NH3 observed up on
quantity of sodium hydroxide heating.
solution, boil in water bath until
the smell of ammonia is
obtained.
2. Test for amides: To the
compound add excess of 2N NaOH a) Red Litmus paper
solution. Boil well and expose a wet turns blue
red litmus paper to the vapours; May be amide
also expose the vapours to glass rod b) Dense white fumes.
dipped in HCI. Cool the solution
and add excess of HCI

3) To a little substance, add a

little quantity of 20% sodium If white crystalline ppt is It indicates presence of aromatic
hydroxide solution and boiled observed amides.
strongly. Then cool the solution and
acidified with dil HCl.

4) Hydroxamic acid test: Boil

approximately 0.1 gm of
substance with 5 ml of water. Magenta colour is observed. Indicates presence of aromatic
Add few drops 3% hydrogen primary amide.
peroxide and 2 drops of
5%FeCl3.Heat the solution.

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5) Biuret test: A little of substance On heating smell of ammonia Indicates presence of diamide.
is heated first gently in a dry test is evolved and violet colour on
tube followed by strong heating. adding CuSO4.
The solid residue is warmed
with 1ml 10% NaOH then
cooled and add one drop of
CuSO4.

ANALYSIS OF NITRO COMPOUNDS

Tests for nitro compounds: 3-4
drops or a small pinch of solid A white solid separates out. The original compound is a nitro-
sample add Tin or Zinc bits and compound
conc HCI. Boil well with shaking
till the yellow compound has
dissolved completely to form a
colourless solution. Filter the hot
solution. Cool the filtrate first under
tap and then in ice. To this, add an
ice-cold solution of sodium nitrite
in water and then an ice cold
solution of β-naphthol in NaOH

Mullikan and Barker’s reaction:

compound and alcohol then add A red dye is formed It is a nitro compound.
concentrated solution of calcium An ash-grey coloured ppt is
chloride and add zinc dust. Boil and formed.
filter. To the Filtrate add Tollen’s
reagent.

Janowsky’s reaction: To the Pale yellow coloured solution It is a mono nitro compound.
Compound add acetone and NaOH
solution. Shake well Violet or pink coloured It is a dinitro compound
solution blue

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ANALYSIS OF ALDEHYDES AND KETONES

Identification of Aldehydes and Ketones:

Test with 2,4-Dinitro Phenyl Hydrazine: Take 2-3ml of 2, 4-dinitro phenyl hydrazine in
a test tube. Add few drops or milligrams of the substance in the reagent. If any yellow oil colour
or yellow precipitate appears. The inference is a carbonyl compound.

Note: There are two kinds of 2, 4 DNP reagents.

a) One for water – soluble compound.
b) One for water insoluble compound.
The yellow oil or organic yellow precipitate formed in the 2,4 DNP hydrazine
corresponding aldehydes or ketones.
c) Some hindered ketones will not give this reaction immediately. So, shake the solution and set
aside for 5-10 minutes. If no precipitate is seen. Hereafter this period, conclusion is not a
carbonyl compound. The hindered carbonyl compound may not give the positive reaction.

Distinguish test for aldehydes and ketones

Schiff’s Test: Take 2-3 ml of Schiff’s reagent in atest tube and add the substance under
investigation to it. If the magenta color (Rose-Pink colour) appears then the substance is
aldehydes. If does not appear, then the substance is ketones.
Note: Schiff’s reagent is a solutionof pararosaniline hydrochloride saturated with
SO2pararosaniline hydrochloride is in solution form with roastingcolour when SO2 gas is passed
into the solution. The dye pararosaniline hydrochloride is oxidized and the color disappears.
When you add aldehyde to this solution. The aldehydes will oxidize the solution to carboxylic
acid and thus the magenta (pink – rose colour) of dye is restored aldehydes are easily oxidizable
substances, ketones are not.

Tollen’s Test: Take 2ml of silver nitrate and 1ml of NaOH solution in a test tube. Add
ammonia solution to this solution. Until, the precipitate formed is completely dissolved (this is a
Tollen’s reagent)

ANALYSIS OF ALDEHYDES

Tollen’s Test: Take 2ml of silver nitrate and few drops of 5% NaOH solution in clean
test tube. Add NH3 solution to this until the precipitate formed is completely dissolved (this is
Tollen’s reagent). Add the substance under investigation (few drops or milligrams), heat on

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water bath. If silver mirror is formed on the inner walls of test tube, the substance is aldehydes. If
no silver mirror is formed, it is ketones.

Note: When silver nitrate is added to sodium hydroxide, a double decomposion reaction
takes place to give AgOH or Ag2O. This precipitate is dissolved by NH3, which forms a complex
with silver ions. The reagent produced is called Tollen’s reagent. i.e, silver ammonium
hydroxide or ammonical silver nitrate. This is reduced by metallic hydroxide, with goes on
oxidation to form acids. The metallic silver gets deposited on the test tube giving the appearance
of mirror. The test tube used for this test must be clean and must not be greasy (wet). If it is
greasy, the metallic silver formed cannot stick to the inner wall of the test tube, but slides down
and sit on the bottom of test tube. So, the test tube must be clean before this test done.

Fehling’s Test: Take in a test tube 1ml of Fehling solution. A (Fehling’s solution-1) and
add 1ml of fehling’s solution-B (fehling solution – 2) To this add few drops of substance, heat on
the flame. The appearance of green, yellow, orange red colour or a brick red precipitate indicates
the presence of aldehydes. If no reduction takes place the substance is ketones.
Note: 1) Fehling’s solution – A is a solution of copper sulphate in water.
2) Fehling’s solution – B is a solution of NaOH and sodium potassium tatarate
(Rochelle’s Salt). When fehling’s solution – A is mixed with Fehlings solution-B, CuSo4 and
sodium hydroxide react to give Na2SO4& Cu(OH)2 (CuSO4 and sodium hydroxide react to give
sodium sulphate and copper hydroxide Cu(OH)2, the Cu(OH)2 is a red ppt, this is kept in a
solution by sodium potassium tatarate, which act as complexing agent.

Iodoform Test: Take a small quantity of substance; add 1 of NaOH solution followed by
Iodine solution. Heat gently on water bath and look for the colour of iodoform. If iodoform is
formed, it is methyl ketone. It is soluble and gives an acetaldehyde.

Identification test for Aldehydes:

Experiment Observation Inference

Formation of Phenyl hydrazone: To 5ml of water, add A derivative of May be an
about 5ml of glacial acetic acid and 0.5 ml of phenyl aldehyde aldehyde
hydrazine. Then add 3 drops of given substance and shake
the test tube for 1-2 minutes.
Presence of
Formation of 2, 4 dinitro phenyl hydrazone (or) Bright orange solid carbonyl
Reaction with 2, 4 DNP: Take 2-3 ml of 2, 4 – DNP separates out compound
reagent in a test tube and add few drops or few mg of
substance to the reagent.

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Formation of semicarbazone Test: Add 0.5 gm of Yellow orange color Presence of

semicarbazine hydrochloride to 5ml of water. Add 0.5 gm crystals are formed carbonyl
of anhydrous sodium acetate and warm gently until clear compound
solution is obtained. Then add a solution of 0.5 ml of the
substance in 2-3ml of alcohol and warm on water bath.

Reduction Ammonical silver nitrate solution or tollen’s

Reagent: Place 5ml of AgNO3 solution in a thoroughly Black ppt May be an
cleaned test tube, add 2-3 drops of dilute NaOH and dilute aldehyde
NH3 solution, drop wise, until the precipitate of Ag2O or
AgOH is almost redissolved, then add 2-3 drops of
substance and warm on waterbath.

Fehling’s Solution: 1ml of Fehling’s solution A+1ml of Yellow to red solid May be an
Fehling’s – B and add few ml or few gm of sample and heat separate out aldehyde
on water bath.

Sodium bisulphate addition compound: shake 1ml of

compound with about 0.5ml of saturated sodium bisulphate White crystalline ppt May be an
solution. The mixture becomes warm and white addition aldehyde
product separates out on cooling, the substance is
regenerated on addition of dil. H2SO4.

KETONES

Characteristics reaction of Ketones:

Experiment Observation Inference

Formation of Phenyl Hydrazone: A florescent ppt May be ketone
To 5ml water, add about 0.5ml of glacial acetic of phenyl
acid and 0.5ml of phenyl hydrazine. Then add 3 dihydrazone is
drops of given substance and shake the mixture produced
for 1 to 2 minutes.

Formation of oximes: Take 1 gm of hydroxyl

amine hydrochloride, 2gms of sodium acetate
and 0.5gm of sample and warm it on water

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insoluble (aldehydes) ketones, with pyridine

catalyst, cool the contents and add 5ml of water
to the cooled residue cool in ice bath and stir
until crystals of oxime separate. Then filter off
and recrystallize from alcohol and benzene.

Formation of 2,4Dinitrophenyl hydrazone: Bright orange May be ketone

Dissolve a few drops of substance in a few ml of solid separates
water or methanol. Adds 2 drops of 2,4 dinitro out
phenyl hydrazine reagent.

Formation of semi carbazone test: Add 0.5 gm

of semi carbazine hydrochloride to 5ml of water.
Add 0.5gm of anhydrous sodium acetate and Yellow orange
warm gently until a clear solution is obtained. color crystals are
Then add 0.5 ml of substance and 2-3ml of formed
alcohol and warm on water bath.

Reduction of Ammonical AgNO3 solution (or)

Tollen’s Reagent:
Place about 5ml of AgNO3 solution in a
thoroughly cleaned test tube. Add 2-3 drops of
dil. NaOH add dil. NH3 solution drop wise until
precipitate of silver oxide is almost redissolved,
then add 2-3 drop of substance and warm on
water bath.

Fehling’s Solution:
1ml of fehling’s solution – A, 1ml of fehling’s
solution – B and few ml/gm of sample are taken
in a test tube and to be heated on water bath.

Sodium Bisulphite addition compound:

Shake 1 ml of compound with about 0.5ml of
saturated sodium bisulphate solution. The
mixture become warm and a white addition
product separation occurring on cooling. The
substance is regenerated on addition of dil.
H2SO4.

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Iodoform Test:
To 0.5 ml methyl ketone, add 3ml of 10% KI
solution and 10ml of freshly prepared. Sodium
hypochlorite solution and mix well.
(OR)
To the sample add excess iodine solution and
sodium hydroxide solution
Nitro Prusside Test:
Add 3ml of freshly prepared sodium nitro
prusside to 0.5ml methyl ketone and add dil.
NaOH solution in excess.

Dinitro Benzene Test:

To 1ml of methyl ketone, add about 0.1 gm of
fine powder dinitro benzene and excess of NaOH
solution and shake well.

Result: The given compound is aliphatic compound and it is a

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ANALYSIS OF AMINES

ANALYSIS OF PRIMARY AMINES

Experiment Observation Inference

1) Carbylamine test: Add few drops of CHCl3 to about A foul or pungent Maybe
0.2gm of the substance and the 2-3ml of alcoholic odour of isonitrile is aliphatic/
KOH. Mix well and warm gently, cool the tube and produced. aromatic
add carefully excess of Conc. HCl. amine

2) Copper sulphate test: Add few drops of 10% blue or blue-green Indicates the
Copper sulphate solution to 0.2gm of the substance. coloration or presence of a
precipitate with the 10 amine.
reagent

3) Diazo-Coupling: Dissolve 0.2 gm of substance in Formation of an Maybe an

0
1ml of Conc. HCl and dilute with about 2ml of cool orange to red dye is aromatic 1
in ice and add few drops of sodium nitrite solution. observed amine
Now add this whole diazonium solution slowly to a
cold solution of 2-Naphthol in an excess of 10% No dye formation Aliphatic
NaOH solution. 10 amine

4) Test with phenol: Add 5ml phenol solution which is Formation of red or Maybe an
dissolved in NaOH solution to the test sample. brown or yellow ppt aromatic 10
amine

5) a. To 0.3 mL or 300 mg of unknown substance in a soluble in base. Presence of

test tube add 5 mL of 10% NaOH solution and 0.4 aliphatic
mL of benzenesulfonyl chloride. Close the test tube primary
with a cork and shake the mixture vigorously. Test amines.
the solution to make sure that it is still alkaline using
litmus paper. Presence of
b. Cool the solution & add 10% HCl solution A precipitate is aliphatic
dropwise. formed. primary
amines.

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ANALYSIS OF SECONDARY AMINES

General Reactions:

Experiment Observation Inference

1) A little amount of substance is heated in a Brown ppt is formed Indicates presence
test tube. of 2o amine.

2) A little amount of substance is treated with formation of yellow Indicates presence

nitrous acid. oil or solid. of 2o amine

3) Formation of Nitroso – amine:

Dissolve 1ml of substance in about 3ml of a green solution turns Presence of 2o
dilute HCl and add NaNO2 solution gradually deep blue on making amine
with shaking until the yellow layer separates it alkaline
out at the bottom of the solution. Transfer
completely into a small separating funnel, add
about 2ml of ether and shake it run off the
lower layer and evaporate the ether to get the
yellow oil of Nitroso derivative.

ANALYSIS OF TERTIARY AMINES

General Reactions:

Experiment Observation Inference

1. Dissolve the substance in dilute White crystalline ppt is Presence of 3o amine
HCl and add potassium Ferro observed
cyanide solution.
2. Formation of p-nitroso
derivative:
Dissolve 0.5 ml of substance in a) a reddish solution is Presence of 3o amine
about 5ml of dilute HCl, cool in ice obtained
and then add carefully 2ml of 20%
NaNO2 solution. Allow it to stand b) A green ppt of p-nitro
for 5 minutes and cooled, then add derivative is formed.
dilute NaOH solution.

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 Pharmaceutical organic chemistry

3. a. Test with nitrous acid: to the The orange-colored Presence of 3o amine

sample add 1 ml of nitrous acid hydrochloride salt of the C-
and 1ml of HCl. nitrosoamine.

b. Treating the C- nitosomine Liberates the blue or green Indicates the presence of
solution with NaOH base. C-nitrosoamine. 3o aromatic amine.

Hinsberg’s test (To different between 1o, 2o& 3o amines).

Take
1 about four drops (or 0.1 gm) of No reaction occurred May be a tertiary or
A the unknown amine in a test tube; add secondary amine
about 0.2 gm of p-toluene sulphonyl
chloride and 5ml of 10% NaOH. Shake
the tube for 5 minutes
If2no reaction appears to occur, heat the i) No precipitation Indicates Tertiary
B reaction mixture on a water bath for 1 appeared amine
minute and cool in ice. ii) Precipitation May be a secondary
produced or primary amine
If3precipitation appears in the alkaline i) precipitation Indicates the presence
C medium and 5ml of water and shake dissolved of primary amine
ii) precipitation not Indicates of presence
dissolved of secondary amine
If4the precipitation dissolves and the Precipitation dissolves Confirms the presence
D solution is clear, acidify with dil. HCl of primary amine

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 Pharmaceutical organic chemistry

ANALYSIS OF ALCOHOLS

Experiment Observation Inference

Specific Test: Evolution of bubbles/ gas is May be an alcohol
To 1ml of Given Test solution add 1 observed
pinch of Sodium Metal in dry test
tube.

Cerric ammonium nitrate Formation of red or yellow May be an alcohol

reagent: to the 1ml of substance add colour.
0.1 gm of ceric ammonium nitrate
reagent.

Ester Formation:
Take 1 ml of substance in A dry Fruity odour is observed May be an alcohol
Test Tube and 0.5 M1 of Acetic
Anhydride or Glacial acetic acid are
added. To It 2 Drops of conc. H2SO4
Is added Mix Thoroughly and Heat
on water Bath and the Mixture Is
poured into a Beaker containing
sodium Bicarbonate Solution

Test with chromic acid:

Add few drops of chromic acid Orange colour solution turn to May be an alcohol
reagent in 1ml of substance the bluish green due to formation
solution will remain clear. Cool the of chromium ion
solution then heats the solution.

Iodoform test
Take 1ml of the substance in the test A characteristic color of May be an alcohol
tube and add 1ml of sodium iodoform is evolved (IF3)
hydroxide solution and few drops of
iodine solution and few drops of
iodine solution. Heat gently on
water bath.

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Distinguish between 1O, 2O& 3 O

alcohols:
Lucas test:
To 2ml of Lucas reagent (anhydrous No visible reaction at room It may be 1o alcohol.
ZnCl2 and HCl) in a dry test tube temperature and cloudy only
and about 0.5ml of alcohol shake on heating.
vigorously a set a side.
Solution turns cloudy in 3–5 It may be 2o alcohol
minutes

solution turns cloudy It may be 3o alcohol

immediately, and/or phases
separate

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 Pharmaceutical organic chemistry

ANALYSIS OF ESTERS

EXPERIMENT OBSERVATION INFERENCE

HYDROXAMIC ACID FROMATION: To a few Violet or deep It is an ester

drops of ester add 0.2gms of Hydroxylamine reddish-brown color
hydrochloride and about 5ml of 10% NaOH and develops
gently boil the mixture for 1-2 minutes. Cool immediately
acidifies with dilute HCl and them adds a few drops
of FeCl3 solution.
ii) Compound + dil NaOH + Phenolphthalein Pink colour It is an ester
and warm with shaking. gradually disappears

iii) Compound + 20% sodium hydroxide A white precipitate It is an ester

solution. Boil for about 10 minutes. Cool is formed
and add excess of dil H2SO4

HYDROLYSIS: Place 2ml of ester in a 250ml

round bottomed flask fitted with reflex condenser. White ppt is formed. Sodium salt of
Add about 20ml of 10% aqueous NaOH solution, 2- (a salt of an acid carboxylic acid
3 porcelain – pieces and reflex gently for 20-30 derivative of ester).
minutes. At the end of this time all oil drops of
unchanged esters should have disappears.
Distillation method is useful in separating the
alcohol.

Department of Pharmaceutical Chemistry.

 Pharmaceutical organic chemistry

ANALYSIS OF CARBOHYDRATES

Molisch’s test: 0.1 g of the A red-cum-violet ring Presence of carbohydrates.

substance is dissolved in appears
1ml of water and treated at the junction of the two
with α-naphthol in alcohol liquids.
(Molisch’s reagent). shake
well &add conc. H2SO4
through the sides of the test
tube.

Fehling’s test: To little Formation of yellow or Presence of reducing sugars

substance 2-3 drops of brownish-red precipitate.
Fehling’s solution A&B are
added and heated.

Benedict’s Test Formation of red, yellow or Presence of reducing sugars

To 2 mL of Benedict’s green colour/precipitate
reagent
add five drops of the test
solution. Boil for five
minutes
in a water bath. Cool the
solution.
.
Barfoed’s Test Formation of brick-red Presence of
To 1 mL of the test solution precipitate monosaccharides.
add about 2 mL of
Barfoed’s reagent. Boil it
for one minute and allow to
stand for a few minutes.
.

Tollen’s test: Silver mirror on the walls of Indicates presence of

A little substance is heated test tube. aldehyde.
with tollens reagent &
heated Black ppt is formed Indicates presence of ketone.
Osazone Test Needle-shaped yellow Presence of glucose, fructose
To 0.5 g of phenylhydrazine crystals and mannose.
hydrochloride add 0.1 g of
sodium acetate and 10 drops mushroom shaped crystals Presence of lactosazone.
of glacial acetic acid. To
this flower-shaped crystals. Presence of maltose.
Department of Pharmaceutical Chemistry.
 Pharmaceutical organic chemistry

mixture add 5 mL of test

solution and heat on a
boiling
water bath for about half an
hour. Allow the tube to cool
slowly and examine the
crystals under a microscope.

Department of Pharmaceutical Chemistry.

 Pharmaceutical organic chemistry

Department of Pharmaceutical Chemistry.

 Pharmaceutical organic chemistry

Department of Pharmaceutical Chemistry.

 Pharmaceutical organic chemistry

Department of Pharmaceutical Chemistry.