Pharmaceutical Organic Chemistry I Lab Manual
Pharmaceutical Organic Chemistry I Lab Manual
Pharmaceutical Organic Chemistry I Lab Manual
LAB MANUAL
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
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.
6. Identification of the unknown compound from the literature using melting point/ boiling
point.
INTRODUCTION
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
Thermometer
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.
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.
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.
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.
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:
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
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:
Percentage yield
Theoretical yield =
Description:
Precaution: Handle bromine carefully as it digests the bones and produces irritation to the eyes
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?
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:
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
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.
Percentage yield:
Percentage yield
Theoretical yield =
Description:
Melting point : 60 to 64 0C
Solubility : soluble in water
Viva Questions:
PREPARATION OF m- DINITROBENZENE
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:
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:
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:
Precaution: m-dinitrobenzene is toxic and do not inhale its vapor. Let it not fall on your skin.
Percentage yield:
Percentage yield
Theoretical yield =
Description:
Melting Point : 89.6 0C
Solubility : freely soluble in chloroform, ethyl acetate
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?
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:
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
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:
Percentage yield
Theoretical yield =
Description:
1. Boiling Point: 126 0C
2. Solubility : miscible in ethanol, soluble in acetone, CHCl3
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:
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:
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
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.
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:
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.
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:
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?
CHEMICALS APPARATUS
Alcohol – 30 ml Conical flask,
10% NaOH solution – 30 ml Measuring cylinder
Iodine solution – q.s Glass rod, Thermometer, Funnel
Principle:
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
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.
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:
Percentage yield
Theoretical yield =
Description:
Melting Point : 119-121 0C
Solubility : soluble in ether, CHCl3, alcohol, slightly soluble in H2O
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:
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:
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.
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.
Percentage yield:
Percentage yield
Theoretical yield =
Description:
Melting point: 120-122 0C
Solubility : slightly soluble in water, freely soluble in ether and ethyl acetate
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 ________
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
SOLUBILITY CHART
Hydrocarbons, nitrohydro-
Insoluble in water, acid carbons, alkyl or aryl halides, esters and
Neutral
and alkali ethers. Higher molecular weight alcohols,
aldehydes and ketones
Lassaignes Test:
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.
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.
ANALYSIS OF PHENOLS
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.
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
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.
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
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.
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
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.
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.
KETONES
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.
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.
ANALYSIS OF AMINES
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
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
General Reactions:
General Reactions:
b. Treating the C- nitosomine Liberates the blue or green Indicates the presence of
solution with NaOH base. C-nitrosoamine. 3o aromatic amine.
ANALYSIS OF ALCOHOLS
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
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.
ANALYSIS OF ESTERS
ANALYSIS OF CARBOHYDRATES