 Experiment No: 01

 Experiment name: Identification of functional group from unknown compound or in

organic compound
 Objectives: Our objective is to identify the functional group present in an organic group
 Test for saturation and unsaturation
 Test for carboxylic acid group
 Test for carbonyl group (aldehyde and ketonic groups)
 Test for phenolic group
 Test for Alcoholic group
 Test for Amino group
 Test for Amide group
 Test for Ether group
 Test for Ester group

 Theory:
Organic compound, any of a large class of chemical compounds in which one or more atoms of
carbon are covalently linked to atoms of other elements, most commonly hydrogen, oxygen, or
nitrogen. Functional Groups, in the field of organic chemistry, are the substituent atoms or
groups

Fig: Qualitative analysis of organic compounds.

of atoms that are attached to specific molecules. A functional group is a group of atoms or bonds
inside a substance that is responsible for the substance’s unique chemical reactions in organic
chemistry. Presence of different of functional groups in organic compounds may be identified by
adding the sample as such or/and a suitable solution to the desire reagent. A reagent is
a compound or mixture added to a system to cause a chemical reaction or test if a reaction
occurs. A reagent may be used to find out whether or not a specific chemical substance is present
by causing a reaction to occur with it.

 Apparatus:
i. Test tube
ii. Test tube holder

 Required Chemicals:
i. Bromine water
ii. Blue litmus paper
iii. 5% NaOH
iv. 5% NaHCO3
v. Ferric chloride solution
vi. DNP
vii. Tollen's reagent
viii. Fehling's solution
ix. Cupric acetate
x. Benzidine hydrochloride
xi. Conc. HCl
xii. Potassium hydroxide (solid)
xiii. Chloroform
xiv. Phenolphthalein
xv. Baeyer’s Reagent
xvi. Ammoniacal cuprous chloride Solution
 Test for identifying Alkane, Alkene and Alkyne
 Procedure:
a) Test with bromine water of CCl4: Add a few drops of bromine water of CCl4 in the given
sample.
b) Test with Byer’s reagent: Add a few drops of Byer’s reagent or alkaline KMnO4 solution
in the given sample.
c) Test with Ammoniacal Cuprous Chloride: Add a few drops of ammoniacal cuprous
chloride solution in the given sample.

Reagent Alkane Alkene Alkyne Result

i) No ii) Red iii) i) The sample contains
reaction Alkane.
color of Decolorizes
ii) Colorless Vicinal
bromine the red color dibromide is
produced, so
disappears of bromine.
sample contains
Add Br2 CH2=CH2 + CH≡CH + Alkene.
water of iii) Colorless 1,1,2,2-
Br2 → 2Br2 →
CCl4 Tetrabromoethane
C2H4Br2 C2H2Br4 is produced, so
the sample
contains Alkyne.
With i) ii) iii) i) The sample
Baeyer’s No reaction Decolorized Decolorized contains Alkane
Reagent the purple the purple ii) Ethylene glycol is
KMnO4 KMnO4. produced thus
CH2=CH2 + CH≡CH + alkene is present
[O] → 4[O] → in the sample.
C2H6O2 (COOH)2 iii) Oxalic acid is
produced thus
Alkyne is present
in the sample.
With No reaction No reaction Red brown
Ammoniaca ppt. is
observed.
l cuprous The sample contains
HC≡CH +
chloride CuCl2 + NH3 Alkyne.
Solution →
CuC≡CCu +
NH4Cl
 Test for Alcohol Group
 Procedure:
a) Test with Sodium metal: In a clean test tube, at first transfer the sample. After that a
small piece of sodium metal that was dipped in the kerosene solution is taken and
dropped into the test tube.
b) Ester test: In a test tube some of the given sample is added with a little amount of
carboxylic acid in the presence of conc. H2SO4.

Sample Reagent and Reaction Observation Result

Test for –OHa) A small piece of Na metal. a) A brisk Alcoholic

2R-OH + 2Na → 2R-O-Na + effervescence is group is

H2(↑) observed due to the present and

liberation of H2 gas. confirmed in

b) Few amount of Carboxylic acid. b) A fruit like smell is the given

R-OH + R-COOH → observed which sample.

R-COOR + H2(↑) indicates the

creation of ester.
 Test for Carboxylic group
 Procedure:
a) Add a few crystals (if solid) or a few drops of liquid to about 1-2 ml of aq. 5% NaOH
solution.
b) Add a few crystals or few drops of liquid to about 1-2 ml of aq. 5% NaHCO3 solution
and shake vigorously.

Sample Reagent and reaction Observation Result

a) 1-2 ml 5% NaOH a) Completely Carboxylic

R-COOH + 5% NaOH → soluble group is present
R-COONa + H2O and confirmed
in the given
Test for -COOH
b) 1-2 ml 5% NaHCO3 b) Completely sample.
R-COOH + 5% NaHCO3 → soluble and a
R-COONa + H2O + CO2 (↑) gas evolves
which turns lime
water milky.

 Test for Carbonyl group (Aldehyde and Ketone)

 Procedure:
a) Add a few crystals (if solid) or a few drops of (if liquid) of the sample to about 1 ml of
DNP reagent in a test tube. Shake well and warm if necessary.
b) To about 1 ml of the freshly produced Fehling’s solution add a few drops of crystals of
the sample. Heat the test tube in a boiling water bath for about half hour.
c) Take 1 ml of silver nitrate solution in a clean test tube and add 2-3 ml of dilute NaOH.
A brown ppt. is formed. Now, dilute ammonia is added dropwise until the brown ppt.
of silver oxide is faded. Then 2-3 ml of the given sample is added and the test tube is
warmed on a water bath for about 5 minutes.
Sample Reagent and Reaction Observation Result

DNP reagent An orange or Carbonyl group is

¿ C=O + H2NNH-C6H3(NO2)2 (2,4) →
Test for yellow ppt. is present in the
¿ C=O -C=NNH-C6H3(NO2)2 (2,4)
observed. given sample.

a) Fehling solution a) A brick red (a)

i) –CHO + Cu(OH)2 → ppt. is i) The sample
-COO- + Cu2O observed is aldehyde
ii) R–C = O–R + Cu(OH)2→ when the ii) The sample
No reaction reagent is ketone.
reacts with
Test for aldehyde.
Aldehyde No reaction
or occurs
Ketone while
reacting
with ketone.
b) Tollen’s Reagent (b)
i) –CHO + 2Ag(NH3)2OH → b) A shinning i) The sample
2Ag(s) + R-C=O-O-NH4+ + silver mirror is aldehyde
H2O + 3NH3 is observed ii) The sample
ii) R–C = O–R + for reaction is ketone.
2Ag(NH3)2OH→ No reaction (i) and no
reaction
occur for
reaction (ii)
 Test for Amide Group:
 Procedure:
a) Hydrolysis: Take the substance in a test tube. Then add about 25% NaOH solution to
this and then heat the solution. While heating we can sense the smell of ammonia gas.
Now cool the solution and neutralize this with the help of dil. HCl acid.
b) Biuret test: Take the sample in a dry test tube and heat it well. Then add about 10%
NaOH solution and mix it well. Now cool the solution and add dil. CuSO4 solution.

Sample Reagent and Reaction Observation Result

a) 25% NaOH a) A smell of NH3 is

R-CONH2 + NaOH + HCl sensed when
NaOH solution is
→ R-COOH + NH3
heated with the
sample. This
indicates the
presence of amide
Amide group
group. Solid acid
precipitation is is present
b) 10% NaOH, few drop of
formed after and
CuSO4.
adding dil. HCl
confirmed in
CH3-CONH2 + Heat → acid.
the given
C2H5N3O2 + NH3
Test for Amides sample.
C2H5N3O2 + CuSO4 →
R-CONH2 b) A violet color is
Biuret forming complex observed when
ion of Cu. CuSO4 is added
which indicates
the presence of
Amide.

 Test for Ester group

 Procedure:
 a) At first take a small amount of sample in a test tube and then add few drops of
phenlophthaline. Then add few drops of NaOH solution. Then heat the solution.
b) Hydroxamic Acid test: Take 2-3 drops of sample in a test tube and add few milligram of
Hydroxylamine hydrochloride solid. Now add 10% NaOH solution and boil the solution in
a water bath for 1-2 minutes. Now acidify the solution by adding dilute HCl acid. Now
add few drops of FeCl3 in the solution.

Sample Reagent and Reaction Observation Result

Test for a) NaOH solution, Phenlophthalin a) Pink color is
R-COO-R R-COO-R + NaOH + observed after
adding
Phenlophthaline → R-COOH +
phenlophthaline
NaOR and the color
slowly
disappeared
after NaOH is
added. The given
sample
contains
Ester.
b) Hydroxylamine hydrochloride solid, b) A violet or
NaOH, dil. HCl, FeCl3 deep red brown
color is
observed.

 Test for ether group:

Procedure:
Feigl's test: Take 1-2 mL of unknown compound in a boiling tube and cover the mouth of the
tube with a filter paper moistened with a mixture of cupric acetate and benzidine hydrochloride
solution. Heat the tube to boiling. Appearance of a deep blue colour indicates presence of ether.
Sample Reagent and reaction Observation Result
Cu(CH3COO)2 (Cupric
Ether acetate), C12H13ClN2
A deep blue ppt is
R-O-R' observed Ether group is present
(Benzidine
hydrochloride)

 Test for phenolic group:

Procedure:
a. Add a few crystals (if solid) or few drops of liquid to about 1-2 mL of aq. 5% NaOH
solution.
b. Add a few Crystals (if solid) or few drops of liquid to about 1-2 mL of aq.5℅ NaHCO 3
solution and shake vigorously.
c. Add few drops of dilute neutral FeCl3 solution to a dilute solution of the compound.
d. Add few drops of bromine water to the dilute solution of the compound.
Sample Reagent and reaction Observation Result
a. Add 5% NaOH a. Completely soluble
b. 5%NaHCO3 b. Insoluble
Phenolic group, c. Add Dilute c. Violet or green Ar-OH is present
Ar-OH neutral FeCl3
Ar-OH +FeCl3 
[Fe-(O-Ar)6]3-

 Test for Amino group :

Procedure:

a. Solubility Test: Take the given organic compound to be tested in a clean test tube. Add 2 to 3
drops of concentrated hydrochloric acid. Shake well the solution and observe the solubility.

b. Carbylamine Test: Take potassium hydroxide solid in a clean test tube and add 2ml of
ethanol to it. Warm the test tube until the potassium hydroxide dissolves completely. To this add
a few drops of chloroform and shake well. To the above mixture add a small amount of the given
organic compound to be tested and warm gently. Observe the odour of the resulting mixture.
Sample Reagent and Observation Result
reaction
a. Conc. HCl a. Completely
soluble
Amino group, C6H5NH2 + HCl → b. An Amino group is
C6H5NH3+Cl- unpleasant present
-NH2 odour Of
b. KOH and isocyanide
chloroform

R-NH2 + 3KOH +
CHCl3 →
RNC(isocyanide) +
3KCl + 3H2O

Precaution:

1.Safety goggles (department approved) must be worn in the lab at all times. Glasses and contact
lenses are not acceptable eye protection. Students who do not follow this rule will be asked to
leave the lab immediately.

2. Never eat or drink in the lab. Food may pick up toxic chemicals.

3. Never inhale fumes or vapours. Use fume hoods for dangerous or irritating chemicals. Always
waft odours toward our nose with our hand.

4. Never taste any chemical. Some chemicals are very corrosive and poisonous in very small
quantities.

5. Never perform an unauthorized experiment and never work in the lab without an instructor in
charge. An accident may happen when mixing simple chemicals.

6. Never remove anything (chemicals, glassware, etc.) from the lab.

7. Must be ware lab coat.

8. Wear appropriate gloves. Gloves are required at all times when we are handling chemicals and
items that have been exposed to chemicals.