ORGANIC QUALITATIVE ANALYSIS

ANALYSIS OF AN ORGANIC COMPOUND

EXPERIMENT OBSERVATIONS INFERENCE

1. Test for Aromaticity

a). Substance is introduced into the flame i. Burns with a smoky Presence of Aromatic
using a Nickel spatula flame compound
ii. Burns with a non-smoky Presence of Aliphatic
flame compound

b). About 5 mg of the substance is added A yellow solution or Presence of Aromatic

to a mixture of 3 drops of conc. Sulphuric precipitate is obtained compound
acid and 3 drops of conc. nitric acid and
warmed on a water bath for about 10
minutes. The solution is poured then into
water.

2. Test for Unsaturation

a). A little of the substance or a drop of i. Decolourisation Presence of

the substance (as the case may be) taken Unsaturation
on a tile. A drop of conc. Bromine water is ii. Decolourisation
Presence of Aniline or
added to it. followed by turbidity
Phenol
formation

b). A pinch of the substance is taken on a Decolourisation Presence of

tile. A drop of Dil. Potassium Unsaturation or easily
permanganate is added on it. oxidisable compounds

3. Solubility Tests
1.
Solubility of the substance in the following solvents is tested

i. Water i. Soluble -Presence of Urea,

carbohydrates, etc.
ii. 5% Sodium bicarbonate ii. Soluble -Presence of Acids
iii. 5% Sodium hydroxide iii. Soluble -Presence of Acids,
Phenols
iv. 5% Hydrochloric acid iv. Soluble -Presence of Amines

4. Action of Litmus
Moistened litmus paper is brought into i. Blue litmus turns Red -Maybe due to acids,
contact with the substance. phenols
ii. Red litmus turns Blue -May be due to amines

iii. Neutral -Presence of

Carbohydrates, esters,
carbonyls, etc.
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5. Action of Sulphuric Acid
About 5 mg of the substance is warmed Substance Chars with the Presence of
with 3 drops of Conc. Sulphuric Acid. smell of burnt sugars Carbohydrates.

6. Sodium Fusion extracts tests (Lassaigne’s Test)

Two small cut pieces of Sodium are fused in a semi micro hard glass tube. About 5 mg of the
substance is added to it and fused again. The tube is cooled to room temperature. Keeping the tube
in slanting position allow one drop of water trickle down. After the first drop has reacted allow the
second drop to trickle down. Likewise add two more drops of water. This is necessary to avoid
spurting of the excess sodium. This is the sodium fusion extract. The following tests are performed
with it.
a). Test for Nitrogen
A drop of the extract is placed on a tile. Prussian Blue colour Presence of Nitrogen
A drop of the concentrated solution of
Ferrous sulphate is added to it. Then it is
treated with a drop Ferric chloride and
acidified with HCl.
6𝑁𝑎𝐶𝑁 + 𝐹𝑒(𝑆𝑂)4 → 𝑁𝑎4 [𝐹𝑒(𝐶𝑁)6 ] + 𝑁𝑎2 (𝑆𝑂)4
2𝑁𝑎4 [𝐹𝑒(𝐶𝑁)6 ] + 4𝐹𝑒𝐶𝑙3 → 𝐹𝑒4 [𝐹𝑒(𝐶𝑁)6 ] + 12𝑁𝑎𝐶𝑙

b). Test for Halogen

A drop of the extract is placed on a tile. A i. Curdy white precipitate Presence of Chlorine
drop of Conc. Nitric acid is added
ii. Pale yellow precipitate Presence of Bromine
followed by a drop of Silver nitrate
solution. iii. Yellow precipitate Presence of Iodine
𝑁𝑎𝑋 + 𝐴𝑔(𝑁𝑂)3 → 𝐴𝑔𝑋 + 𝑁𝑎(𝑁𝑂)3
*If Halogen present
About 5 mg of the substance is warmed i. Immediate formation of Halogen is present in
with 3 drops of alcoholic Silver nitrate on a precipitate the Side chain
a water bath and acidified with a drop of
ii. No precipitate Halogen is present in
Conc. Nitric acid.
the Aromatic nucleus
𝐶6 𝐻5 𝐶𝐻2 𝑋 + 𝐴𝑔(𝑁𝑂)3 → 𝐴𝑔𝑋
𝐶6 𝐻5 𝑋 + 𝐴𝑔(𝑁𝑂)3 → 𝑁𝑜 𝑅𝑒𝑎𝑐𝑡𝑖𝑜𝑛

c). Test for Sulphur

i. A drop of the extract is mixed with a Violet Colouration Presence of Sulphur
drop of Sodium nitroprusside on a tile.
𝑁𝑎2 𝑆 +𝑁𝑎2 [𝐹𝑒(𝐶𝑁)5 𝑁𝑂] → 𝑁𝑎2 [𝐹𝑒(𝐶𝑁)5 𝑁𝑂𝑆]

ii. A drop of the extract is tested with a Black precipitate Presence of Sulphur
drop of Lead acetate solution on a tile.
(𝐶𝐻3 𝐶𝑂𝑂)2 𝑃𝑏 +𝑁𝑎2 𝑆 → 𝑃𝑏𝑆 + 2𝐶𝐻3 𝐶𝑂𝑂𝑁𝑎

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 EXPERIMENT OBSERVATIONS INFERENCE

7. Test for Functional groups

a). Test for Carboxylic acid
i. About 0.025 g of the substance is treated Brisk effervescence Presence of Carboxylic
with a drop of highly saturated Sodium acid
bicarbonate solution on a tile.
𝑅 − 𝐶𝑂𝑂𝐻 +𝑁𝑎𝐻𝐶𝑂3 → 𝑅 − 𝐶𝑂𝑂𝑁𝑎 + 𝐶𝑂2 + 𝐻2 𝑂

ii. A drop of neutral ferric chloride i. Violet colour -Presence of Salicylic

solution is added to a little of the acid (or) Phenol
substance.
ii. Flesh colour -Acid like Phthalic

b). Test for Dicarboxylic acid

About 5 mg of the substance is heated Green Fluorescence
Presence of
with equal amount of Resorcinol and 2 Dicarboxylic acid
drops of Conc. Sulphuric acid in a semi
micro test tube. The solution is cooled and
diluted with about 1 ml of water. A drop of
this solution is mixed with a drop of 10 %
Sodium hydroxide on a tile.

c). Test for Phenol

1. Neutral Ferric chloride test

A drop of the substance is mixed with a Violet colouration Presence of Phenol

drop of neutral Ferric chloride solution on
a tile.
6𝐶6 𝐻5 − 𝑂𝐻 + 𝐹𝑒𝐶𝑙3 → 𝐻3 [𝐹𝑒(𝑂𝐶6 𝐻5 )6 ] + 𝐻𝐶𝑙

2. Phthalein Reaction
About 5 mg of the substance is heated Red, blue or green colour Presence of Phenol
with about 10 mg of Phthalic anhydride is produced
and a drop of Conc. Sulphuric acid in a
semi micro tube. The solution is cooled
and diluted with about 1 ml of water. A
drop of the solution is mixed with a drop
of 50 % Sodium hydroxide solution on a
tile.

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3. Liebermann Reaction
About 5 mg of the substance is heated A bluish green colour is Presence of Phenol
with Sodium nitrite and Conc. Sulphuric produced
acid in a semi micro tube. It is cooled and
diluted with about 1 ml of water. A drop of
it is placed on a tile and mixed with a drop
of 10 % Sodium hydroxide solution.

d). Test for -Naphthol (Molisch’s test)

A drop of the alcoholic solution of the Violet colouration Presence of -Naphthol
substance is placed on a tile. It is mixed
with a drop of an aqueous solution of
Glucose. To this add a drop of Conc.
Sulphuric acid.

e). Test for -Naphthol

A drop of aniline is placed on a tile. A drop A scarlet red eye Presence of -Naphthol
of Dil. Hydrochloric acid or Sulphuric acid
is added to it. A drop of Sodium nitrite
solution is added. Then a drop of alkaline
solution of the substance is added to it.

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 EXPERIMENT OBSERVATIONS INFERENCE

f). Test for Primary Amines

1. Dye Test: A drop of the substance Red azo dye Presence of Aromatic
is placed on a tile. A drop of Dil. Primary amine
Hydrochloric acid is added to it. A drop
of Sodium nitrite solution followed by a
drop of -Naphthol dissolved in 25 %
Sodium hydroxide is added to it.
2. Carbylamine reaction: To a little An offensive smell is Presence of Primary
of the substance few drops of produced amine
Chloroform and about 2ml of alcoholic
potash are added and warmed.

g). Test for Secondary Amines

2 drops of substance, 2 drops of Dil. A bluish green colour is Presence of Secondary

Hydrochloric acid and 2 drops of produced Amine
saturated solution of Sodium nitrite are
taken in a semi micro tube. A drop of
Conc. Sulphuric acid and a drop of Phenol
are added. Heated for a minute. A drop of
this solution is mixed with a drop of Dil.
Sodium hydroxide on a tile.

h). Test for Tertiary Amines: Malachite green test

2 drops of Benzaldehyde and 4 drops of Malachite green dye is Presence of Tertiary

the substance are heated with 1 drop of obtained Amine
Conc. Sulphuric acid, then it is heated
with about 50 mg of lead dioxide. A drop
of this solution is mixed with a drop of Dil.
Hydrochloric acid on a tile.

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 EXPERIMENT OBSERVATIONS INFERENCE

i). Test for Nitro group

1. Reduction to Amines: About 3
Red azo dye Presence of Nitro
drops of the substance is reduced with 3
compound
drops of Conc. Hydrochloric acid and
metallic Tin or Zinc in a semi micro tube,
by heating for about 5 minutes, with a
drop of the supernatant liquid dye test is
performed on a tile (as under test for
amines).

2. Mulliken-Barker’s test: About 3 Black precipitate Presence of Nitro

drops of the substance and 3 drops of compound
alcohol are boiled with 3 drops of
Calcium chloride solution and a pinch of
Zinc dust. Heated to boiling and cooled.
A drop of it is mixed with a drop of
Tollen’s reagent on a tile.

j). Test for Anilide

1. Action of NaOH: A little of the Separation of oil and
Presence of Anilide
substance with NaOH solution. formation of an emulsion

2. About 10 mg of the substance is

heated strongly in a dry semi micro tube Scarlet red dye Presence of Anilide
with Soda lime. The resulting vapour is
collected in another semi micro tube
containing 5 drops of Dil. Hydrochloric
acid. With a drop of the acid solution
Dye test is performed.
1.

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 EXPERIMENT OBSERVATIONS INFERENCE

k). Test for Amides

1. Biuret Test: About 5 mg of the
substance is heated in a dry semi micro A violet colouration is Presence of diamide
tube to its melting point. Cooled, produced
dissolved the residue in 4 drops of water.
A drop of it is mixed with a drop of Dil.
Copper sulphate and a drop of Dil.
Sodium hydroxide solution on a tile
∆ 𝐶𝑢𝑆𝑂4
2𝑁𝐻2 𝐶𝑂𝑁𝐻2 → 𝑁𝐻2 𝐶𝑂𝑁𝐻𝐶𝑂𝑁𝐻2 → 𝑉𝑖𝑜𝑙𝑒𝑡 𝑐𝑜𝑙𝑜𝑢𝑟𝑒𝑑 𝑐𝑜𝑚𝑝𝑙𝑒𝑥
(Biuret)

2. Test for thiourea

a. About 5 mg of the substance is A black or brown Presence of thiourea
heated with 5 drops of Dil. Sodium precipitate
hydroxide solution, cooled. A drop
of it is mixed with a drop of Lead
acetate solution on a tile.

1)𝑁𝑎𝑂𝐻 2)𝐿𝑒𝑎𝑑 𝑎𝑐𝑒𝑡𝑎𝑡𝑒

𝑁𝐻2 𝐶𝑆𝑁𝐻2 → 𝑃𝑏𝑆

b. 0.005 gm of the substance is heated Blood red colouration Presence of thiourea

in a dry semi micro tube until it
melts. Cooled and then dissolved the
residue in 4 drops of water. A drop of
this solution is mixed with a drop of
neutral Ferric chloride solution on a
tile.

3. Test for Aromatic Monoamide

White precipitate Presence of monoamide
About 5 mg of the substance is heated
with 5 drops of 25 % Sodium hydroxide
solution, cooled. A drop of it is mixed with
a drop of Conc. Hydrochloric acid.
𝑐𝑜𝑛𝑐.𝐻𝐶𝑙
𝐶6 𝐻5 𝐶𝑂𝑁𝐻2 + 𝑁𝑎𝑂𝐻 → 𝐶6 𝐻5 𝐶𝑂𝑂𝐻

l). Test for Carbohydrates

1. Molisch’s Test: A drop of an
alcoholic solution of -Naphthol is A deep violet colouration Presence of
mixed with a drop of aqueous solution of Carbohydrates
the substance on a tile. This mixed
solution is allowed to come in contact
with a drop of Conc. Sulphuric acid.

(Reaction as under test for α-Naphthol)

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2. Osazone Test: About 0.5g of

substance is dissolved in water to get a Yellow crystals are Presence of
saturated solution. It is mixed with obtained Carbohydrates
about 2 l of freshly prepared Phenyl
hydrazine reagent and the mixture is
heated on a water bath occational.

m). Test for Aldehydes

1. Borsche’s reagent Test: A drop Red – Orange precipitate Presence of aldehydes
of the substance (if it is a liquid) or a
drop of an alcoholic solution of the
substance is placed on a tile. A drop of a
Conc. solution of 2,4-Dinitro
Phenylhydrazine (Borsche’s reagent) is
added to it.

2. Schiff’s reagent Test: A drop of Pink colouration Presence of aldehydes

the substance is placed on a tile. A drop
of Schiff’s reagent is mixed with it.

3. Fehling’s Test: 3 drops or about 5 Red precipitate Presence of Aldehydes

mg of the substance is mixed with 2 drops or reducing sugars
of Fehling A and 2 drops of Fehling B
solutions in a semi micro test tube. It is
heated in a water bath for about 5
minutes.
𝑅 − 𝐶𝐻𝑂 + 2𝐶𝑢2+ + 5𝑂𝐻 − → 𝐶𝑢2 𝑂 + 𝑅𝐶𝑂𝑂− + 3𝐻2 𝑂
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n). Test for Ketones

1. Borsche’s reagent Test is A yellow precipitate is

conducted Presence of ketones
obtained

2. Iodoform test is conducted:

Iodine solution is added to small amount A pale-yellow precipitate is Presence of ketone
of the substance, followed by just formed group containing
enough sodium hydroxide solution to CH3CO-group.
remove the colour of the iodine. Warm
gently if necessary.
𝐶𝐻3 𝐶𝑂 − 𝐶6 𝐻5 + 3𝐼2 + 4𝑂𝐻 − → 𝐶𝐻𝐼3 + 𝐶6 𝐻5 𝐶𝑂𝑂− + 3𝐼 − + 3𝐻2 𝑂

o). Test for Esters

1. About 3 drops of the substance is
heated with 3 drops of methanolic Violet colour Presence of Esters
solution of Hydroxylamine
hydrochloride, 2 drops of 50 % Sodium
hydroxide and cooled. A drop of it is
placed on a tile. A drop of Dil.
Hydrochloric acid is added to it. Then a
drop of neutral Ferric chloride is added
to it.

2. 2 drops of the substance are

heated with 2 drops of 50 % Sodium White precipitate Presence of Esters
hydroxide solution, till it is dissolved,
cooled and then added 4 drops of 50 %
Hydrochloric acid.
𝐻𝐶𝑙
𝐶6 𝐻5 𝐶𝑂𝑂𝐶𝐻3 + 𝑁𝑎𝑂𝐻 → 𝐶6 𝐻5 𝐶𝑂𝑂𝐻 + 𝐶𝐻3 𝑂𝐻 + 𝑁𝑎𝐶𝑙

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p). Test for Hydrocarbons

2. Nitration: About 5 mg of the
substance is added to a mixture of 3 Yellow solid obtained Presence of
drops of conc. Sulphuric acid and 3 Hydrocarbon
drops of conc. nitric acid and warmed on
a water bath for about 10 minutes. The
solution is poured then into water.

1. Picrate test: About 0.5 g of the

substance is dissolved in hot benzene is Red or yellow precipitate Presence of polynuclear
added to picric acid dissolved in Hydrocarbon
benzene and mixed well.

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 DERIVATIVE PREPARATION
1. Derivative of aromatic polynuclear hydrocarbon: Picrate derivative
Dissolve 0.2g of the aromatic polynuclear hydrocarbon in the minimum of hot benzene. Add this
solution to 2ml of saturated solution of picric acid in benzene. Warm this mixture and cool it to allow
the product to crystallize. Filter the product and wash with a little ether. Recrystallize this product from
benzene. Determine the melting point.
2. Derivative of aromatic halogen compound: Nitro derivative
[For a halogen compound having halogen in the nucleus] Add slowly 1g or 1ml of the nuclear halogen
compound to 4 ml of concentrated H2SO4 taken in a test tube. Add continuously 4 ml of fuming HNO3
in drops. Heat the mixture on a boiling water bath for 10 to 15 minutes. Pour the content of the test tub
into 25 ml of ice-cold water taken in a small beaker. Filter off the precipitated nitro derivative, wash it
with a little water and dry Recrystallize the product from dilute ethanol. Determine the melting point.
3. Derivative of Aldehyde and Ketone: 2,4-Dinitrophenylhydrazone
Dissolve about 500 mg of carbonyl compound (aldehyde/ketone) in the minimum volume of methanol.
Add this solution to about 5ml of 2,4-Dinitrophenylhydrazine (Borsche’s reagent). Shake the solution
vigorously for few minutes. If no solid separates out in 10 minutes, dilute the solution with dilute H2SO4.
Filter off 2,4-Dinitrophenylhydrazone and wash it with a little methanol. Recrystallize the product from
dilute ethanol. Determine the melting point.
4. Derivative of Ester: Acid derivative
About 1 g of the substance is mixed with 10ml of 20% NaOH and boiled under reflux for about an hour.
The product is then acidified with concentrated HCl. The acid formed is separated.
5. Derivative of Salicylic acid: Nitro derivative
Add slowly 1g compound to 4 ml of concentrated H2SO4 taken in a test tube. Add continuously 4 ml of
fuming HNO3 in drops. Heat the mixture on a boiling water bath for 10 to 15 minutes. Pour the content
of the test tub into 25 ml of ice-cold water taken in a small beaker. Filter off the precipitated nitro
derivative, wash it with a little water and dry Recrystallize the product from dilute ethanol. Determine
the melting point.
6. Derivative of Thiourea: S-Benzylthiouronium chloride derivative
Mix 1 ml of Benzyl chloride and 0.75g of thiourea followed by 2 ml of rectified spirit. It is heated on a
water bath for half an hour and it is cooled in ice. The colourless product formed is separated and dried.
Determine the melting point.
7. Derivative of Phenol: Benzoyl derivative
To about 1g of phenol taken in a conical flak, add 5ml of benzoyl chloride and 20ml of 10% NaOH.
Shaken well after closing the conical flask with cork. Then the mixture is poured into cold water.
8. Derivative of Amines:
a. Benzoylation: The amine is treated with a little excess of NaOH (5ml) in a boiling tube. 1ml of
Benzoyl chloride is added in small amounts with constant shaking. The boiling tube is tightly corked
and shaken well for 5 – 10 minutes. The solid is filtered and washed well with water to remove the
excess alkali. It is recrystallized from dilute alcohol. Melting point is determined.
b. Bromo derivative: 0.5g of the substance is dissolved in 0.5ml of glacial acetic acid. To this is added
bromine in glacial acetic acid till the colour of bromine persists. After 15 minutes the mixture is
poured into cold water. It is filtered and recrystallized from alcohol.
9. Derivative of Acids:
a. Anilide derivative
About 0.4g substance and pure aniline are taken in a dry test tube. The mixture is boiled under reflux
for about an hour, cooled and poured in excess of dilute HCl. It is filtered at the pump, washed with
water and dried. It is the recrystallized from dilute alcohol. Determine the melting point.
b. S-Benzylthiuronium salt derivative

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10. Derivative of aromatic Diamide
To a saturated solution of the amide in water, concentrated HNO3 is added drop by drop till a precipitated
is formed. It is recrystallized from alcohol.

11. Derivatives of Monoamide: Acid Derivative

0.5g of the substance is hydrolysed by heating with 5 ml of 10% NaOH and the acid is isolated after
acidification with dil. HCl. It is recrystallized from alcohol.
12. Derivative of Anilide:
a. Bromo derivative
b. Nitro derivative
13. Derivative of Carbohydrates: Osazone derivative
To 5ml of a 1% solution of the compound, a mixture of 0.1g of Phenylhydrazine hydrochloride and
0.25g of NaOH are added. Then 3 drops of glacial acetic acid are also added. The mixture is heated on
a boiling water bath for 15 minutes. The osazone is recrystallized from dilute alcohol.
14. Derivative of Nitro compound
a. Nitration
b. Reduction: [For mono nitro compounds] Compound is reduced to corresponding primary amine.
Then (1) coupled with Phenol to form azodye (2) benzoylation is conducted.

PREPARATION OF SOME COMMON REAGENTS USED IN ORGANIC ANALYSIS

1. Borsche’s reagent: Dissolve 1g of 2,4-dinitrophenylhydrazine in 5ml conc. H2SO4. Add this solution
carefully to 95ml of rectified spirit with stirring. Allow to stand and filter.
2. Schiff’s reagent: Dissolve 0.25g of para-rosaniline hydrochloride (Fuschine) in 25ml of distilled water
and saturate it with SO2 till the solution becomes colourless. Dilute the solution with distilled water to
250 ml and keep in a tightly stoppered amber coloured bottle.
3. Neutral ferric chloride: Dissolve 1 g of ferric chloride in 100ml of water. Add saturated sodium
carbonate solution in small amounts till a slight turbidity persists which does not disappear even on
shaking. Remove the precipitate by filtration and use the filtrate.
4. Tollen’s reagent: Take 2ml of 0.1N silver nitrate solution and add to it a drop of 2N NaOH solution.
Add 2N NH4OH solution dropwise until the turbidity (reddish brown precipitate of silver oxide) that
appears just re-dissolves to yield a clear solution.
5. Fehling’s solution:
*Fehling’s solution A: Dissolve 17.5g of crystalline copper sulphate in 250ml of water.
*Fehling’s solution B: Dissolve 87g of Rochelle’s salt (Sodium potassium tartrate) and 37g of sodium
hydroxide separately in about 75 – 100ml of water each, mix the two solutions and then dilute to 250ml.
6. Phenolphthalein: Dissolve 5g of phenolphthalein in 500ml of ethanol and add 500ml of water with
constant stirring. Filter if necessary.

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