SCHEME OF ORGANIC ANALYSIS

The Scheme of Analysis may be divided into five parts

1.Preliminary tests
2.Detection of elements
3.Detection of Characteristic groups
4.Confirmatory tests
5. Determination of melting point (in case of solid)
6. Confirmation by preparing a solid derivative for identifying the organic compound.

Preliminary tests (gives indication of which type of compound it is)

Experiment
1. Colour and appearance
thesubstance are noted

Observation
of (a) Colourless

2. Odour is noted

Inference
Hydrocarbon, aldehydes, ketones,
acids esters etc may be present

(b) Yellow

Aromatic nitro compounds may

be present

(c) Brown or black

Phenol or amine may be present

(a) Pleasant fruity

Presence of ester

(b) Fishy or ammoniacal

Presence of amines

(c) Kerosene like smell

Presence of hydrocarbon

(d) Bitter almond smell

Presence of benzaldehyde or
nitrobenzene

(e) Pungent

Halogen
present

(f) Carbolic

Presence of phenol

Soluble

Presence of sugars, lower

aliphaticalcohols,
aldehydes,
ketones andesters.

Insoluble

Presence
of
aromatic
hydrocarbons, amines, phenols,

compounds

may

be

3. Solubility is noted.
A little of the given compound is
shaken with the followings olvents
(a) Cold water

(b) Sodium hydroxide

higher
esters.

(c) Dil. Hydrochloric acid

aldehydes,ketones

and

Soluble
in
sodium Presence of acids or phenols.
hydroxide and reappears
as turbidity on adding
excess of dil. HCl.
Soluble and reappears as Presence of amines
turbidity on adding excess
of NaOH solution

II. Detection of Elements

Lassaignes Test
A small piece of metallic sodium is placed in an ignition tube and melted by gentle heating. Then the
tube is removed from the flame and a small quantity of the substance is rapidly added to the melted
sodium. It is again heated gently to complete the reaction and then strongly. When the ignition tube is
red hot it is plunged into distilled water taken in a china dish. The tube breaks and any residual sodium
react with water. The broken ignition tube is ground well with the bottom of a boiling tube. The mixture
is boiled well and filtered and the filtrate is known as the sodium fusion extract. If necessary, this step is
repeated. The following tests are done with the extract.
Experiment
Observation
Inference
To one portion of the sodium
fusion extract half of its volume
of freshly prepared ferrous
sulphate solution(about 50 mg
in min. vol. of water) is added,
heated gently and acidified with
dil.H2SO4.
Another portion of the extract is
acidified with excess dil. Nitric
acid (approx. equal vol.), boiled
to half the volume, cooled and
silver nirate solution is added

A blue or green colouration or Presence of Nitrogen

precipitate is obtained

(a) White curdy

soluble in ammonia

precipitate Chlorine is present

(b) Yellowish white precipitate Bromine is present

sparingly soluble in ammonia

(c) yellow precipitate insolublein Iodine is present

ammonia
To the third portion of the Violet colouration
Sulphur is present
extract a few drops of freshly
prepared sodium nitroprusside
solution is added

III DETECTION OF CHARACTERISTIC GROUPS

1. Test to find whether aliphatic or aromatic
Ignition test.
A small quantity of the
substance is ignited on nickel
spatula

(a) Burnt with a non-smoky

flame
(b) Burnt with a smoky
luminous flame
2. Test to find out whether unsaturated or saturated
i) Action of dilute potassium
permanganate: A little of the (a) Immediate decolourisation
substance is shaken with water
and one or two drops of dil. b) Slow decolourisation
potassium
permanganate
solution

Presence of aliphatic substance

ii) Action of bromine water: A a) Decolourisation without the

little of the substance is
formation of a precipitate.
dissolved in suitable solvent
(alcohol/ water) then a little
b) Very slow decolurisation
bromine water is added
c) No decolurisation

Presence
compound.

3. Action of sodium hydroxide solution:

A little of the substance isboiled (a) Ammonia is evolved
with dilute sodium hydroxide
solution
(b) substance dissolved

Presence of aromatic substance

Presence
of
unsaturated
compound
Presence of easily oxidizable
substance like phenol, nitro
phenol,
amines,
belnzaldehyde,etc.
of

unsaturated

Presence of easily oxidizable gr.

Presence
compound.

of

saturated

Presence of amide

Presence of acidic substances

like acids phenols and their
derivatives
(c) Separation of oil or formation Presence of anilides
of an emulsion
(d)Solution turns deep yellow in Presence of nitrophenols
colour
4. Action of sodium bicarbonate:
To one mL solution of sodium Brisk
effervescence
of Presence of acids
bicarbonate solution little of the carbondioxide
substance is added
5. Action of ferric chloride solution:
To a little of the substance in (a) Violet or green colour
Presence of phenolic OH
water or alcohol (if not soluable
in water) a few drops of neutral (b) A flocculent white precipitate Presence of -naphthol
ferricchloride is added

(c) green colour changing to a Presence of -naphthol

white precipitate

6. Action of Borsches reagent (2,4-DNP test)

To one mL of Borsches reagent
(2,4-DNP) a little of the
substance is added and heated A yellowish orange precipitate is
over a few minutes (if required). obtained
Cooled and little water is added.
7. Action of Fehlings solution
Fehlingssolution A and Fehlings
solution B aremixed in equal
volumes. To 1 mL of this reagent Reddish
brown
precipitate
a little of the organic compound isformed
is added and heated on a boling
water bath

Presence
ketones

of

aldehydes

or

Presence
of
aldehydes,
polyhydric phenols, and reducing
sugars.

8. Action of Tollens reagent (perform if required)

A little of the substance is added (a) Bright silver mirror is formed
to about 2ml Tollens reagent in
a clean test tube and heated in a
boiling water bath
(b) Black or brown precipitate

Presence of aldehydes, reducing

sugars such as glucose, fructose,
maltose etc.
Presence of polyhydric phenol

9. Action of Molishs reagent (perform only if you get hint of presence of sugar)
To a solution of substance in
water added a few drops of
alcoholic solution of -naphthol. Violet ring is formed at the Presence of Carbohydrates.
Then added about 1mL of junction
con.H2SO4 along the sides of the
test tube without disturbing

IV Confirmatory Tests
A. If nitrogen is present is present, the following tests are conducted. Besides the following tests for
those groups for which indications are got are also done.
1 Action of sodium hydroxide solution: (a) Ammonia is evolved
Presence of amides
A little of the substance heated with
sodium hydroxide
(b) Separation of oil and Presence of anilides
formation of an emulsion

2. Action of nitrous acid: A little of the

substance is dissolved in dilute
hydrochloric acid, cooled in ice
waterand a 10% solution of sodium
nitrite isadded with shaking till it is
slightly inexcess

(a) Liberation of nitrogen with Presence

of
the formation of alcohol.
primary amines.
(b) Separation of an yellow oil.

Presence
of
secondary
amines
(c) Reddish brown solution Presence of tertiaryamines
isobtained.

3. With the solution obtained above

the following tests are done.
To one portion of the solution A scarlet
analkaline solution of -naphthol isformed.
isadded.
(ii) A portion of the solution
isextracted with ether. The ether Blue
or
extract is washed with sodium isobtained.
hydroxide solution and then with
water. The ether is evaporated off and
Liebermanns nitroso reaction is
conducted with the residual oil.
(iii) To another portion, dilute
sodiumhydroxide solution is added
and then shaken with little ether.
(perform last two tests only if primary
NH2 gr. is absent)
4. Mulliken and Barkers reaction. A
little of the substance is dissolved in
alcohol (MeOH). A few drops of
calcium chloride solution is added and
pinch of zinc dust. Boiled for five
minutes, cooled and filtered into a
tube containing Tollens reagent.
5. Reduction of nitro group to amino
group (perform this test if NH2 gr. is
not present): A little of the substance
is treated with few ml of dilute
hydrochloric acid (6N) and a pinch of
zinc dust. Heated for some time and
filtered.
To one portion of the filtrate dil.
hydrochloric acid is added, cooled in
ice and sodium nitrite solution is
added in excess. Then alkaline naphthol solution is added.

aliphatic

red

green

Ether
layer
deepgreen.

precipitate Presence
of
primary amines.

aromatic

solution Presence
secondaryamines.

of

becomes Presence of tertiaryamines.

Bright silver mirror or black Presence of nito group.

precipitate is obtained.

A scarlet precipitate isobtained

Presence of aromatic nitro

group.

B. If halogen is present, the following tests are conducted. Besides the following tests, tests for those
groups for which indications are got also done
Action of alcoholic silver nitrate. (a) Precipitate of silver halide is Presence of halogen in the side
To alittle of the substance 2 ml obtained
chain
of alcoholicsilver nitrate solution
is added andwarmed gently
(b) No precipitate of silver halide Presence of halogen in the ring.

C If sulphur is present, the following tests are conducted. Besides the following tests, the tests for those
groups which indications are got also done.
1 Action of alcoholic sodium
hydroxide: To a little of the
substance 2 ml of alcoholic
sodium hydroxide (one bead in 1
ml alcohol) solution is added and
warmed gently.
2 Fusion with alkali. A little of
the substance is fused with
sodium hydroxide dissolved in
water and dil. hydrochloric acid
is added

Ammonia is evolved

Presence
of
sulphonamide

thiourea

(a) Hydrogen sulphide is evolved

Presence of thiourea

or

(b) Sulphur dioxide is evolved Presence of sulphonic acid

with the formation of phenol
(c) No phenol is formed but Presence of aminosulphonic acid
precipitate of barium sulphate
when barium chloride is added
(d) Ammonia is evolved during Presence of sulphonamide
fusion. No phenol is formed.
Sulfur dioxide is evolved on
adding acid

D. If nitrogen, halogens and sulphur are absent, tests for the following groups for which indications are
got, are done.
I.Aldehydes
1 Borsches reagent test (2,4-DNP An yellow precipitate is obtained.
test) is conducted.

Presence of aldehydes or
ketone.

2 Fehllings
conducted.

Presence of aldehydes.

solution

test

is Red precipitate is obtained.

3 Tollens reagent test is conducted Bright mirror or black precipitate is Presence of aldehydes
(if required)
obtained.

6 Semicarbazide test (if required): White crystalline

Dissolved 0.2 g of semicarbazide obtained.
hydrochloride in 2 ml of water and
added 0.2g of anhydrous sodium
acetate. It is warmed to get a
solution. Then added a small
quantity of the substance and
warmed on a water bath.

precipitate

is Presence of aldehydes.

II. Ketones.

1 Borsches reagent test is An yellow precipitate is obtained

Presence of ketones.
conducted.
2. Semicarbazide test is conducted.
White crystalline precipitate is Presence of ketones.
obtained
III.Acids
1 Tested with sodium bicarbonate Effervescence
solution.
2 Ester formation test is conducted:
About 0.2 g of the substance (one
full spatula) is heated gently with Pleasant fruity smell.
about 1 ml of ethanol (CAUTION!
dont bring the mouth of the test
tube closer to bare flame, it will
catch fire) and few a drops of conc.
sulphuric acid for about 1 minute.
Cooled and poured into a few ml of
water in test-tube.
IV.Phenols
1 Neutral ferric chloride solution test (a) Violet blue or green colour.
is conducted. A little of the
substance is treated with neutral (b) A flocculent white precipitate.
ferric chloride solution

Presence of acids

Presence of acids

Presence of phenol
Presence of-naphthol.

2 Phthalein fusion reaction:

About 2 drops of melted phenol is Red, bluish-purple, blue green Presence of phenol.
mixed with a small quantity of fluorescene, green or very faint
phthalic anhydride in a dry test- green colouration.
tube. 2 drops of conc. H2SO4 are
added. The mixture is heated at

about 150 oC for 2 min. Cooled and

exess of 10 %NaOH solution is
added.
3 Azo-dye formation reaction:
Dissolved 2 drops of aniline in 1 ml Orange, scarlet,dark red,brownish Presence of phenol.
dil. HCl and well cooled in ice. A red solution or precipitate is
fewdrops of saturated NaNO2
obtained.
solution are added. The diazonium
solution thus obtained is added to a
well cooled solution of phenol in
aqueous NaOH solution.
VI. Esters.

1 Hydrolysis. A little of the substance White precipitate is formed.

Presence of ester.
is refluxed with concentrated
solution of sodium hydroxide (2
beads in 2 ml of water) and then
acidified with conc. Hydrochloric
acid.
2 Hydroxamic acid formation.
To a few drops of the substance, A violet or a deep red-brown colour Presence of ester
added 0.2g of hydroxylamine developed immediately
hydrochloride and about 5ml of 10%
sodium hydroxide solution and the
mixture gently boiled for 2 minutes.
Cooled and acidified with dilute
hydrochloric acid and then added a
few drops of ferric chloride solution.
VII Carbohydrates.
1 Concentrated sulphuric acid test is
conducted. Warmed a little of the
substance with conc. sulphuric acid.
2 Molischs test is conducted.
3 Treated with
Tollens reagent
4 Fehlings solution test
is conducted: Warmed
Fehlings solution.
5 Osazone test is conducted

Charring with smell of burnt sugar

A deep violet ring is formed.

Bright silver mirror or
precipitate.

with Red precipitate is formed.

Presence of carbohydrate.

Presence of carbohydrate.
black Presence of reducin gsugar.

Presence of reducing sugar.

Yellow crystals are formed.

Presence of carbohydrate.

Kerosene like smell observed.

Presence of hydrocarbons

VIII. Hydrocarbons.
1 Odour is noted

2 performed test for all functional None of the above tests positive
groups
4 Picrate test is conducted :
Saturated solutions of naphthalene Red or yellow precipitate.
and picric acid, both in benzene are
prepared separately.These two
solutions are mixed in a watchglass
and allowed to evaporate.

Presence of hydrocarbons

Presence of
hydrocarbons.

polynuclear

5. Determine MP of the O.S.

6. Confirmation by preparing a solid derivative.
The final step in the analysis of a sample organic compound is the preparation of a suitable solid
derivative. PLEASE CONSULT INSTRUCTORS BEFORE PREPARATION OF DERIVATIVES.
Preparation of Derivatives
1. Derivatives for Aldehydes and Ketones.
a) Semicarbazone derivative:
Procedure: Dissolve a small amount of the compound (approx. 200 mg) in ~2 mL 95%
Ethanol. Add aqueous solutions of 0.5 g semicarbazide hydrochloride (in 0.5 ml) followed by
aqueous sodium acetate (0.7 g in 0.5 ml). Mix the liquids thoroughly. If and only if two liquid
layers form, add 95% ethanol dropwise while shaking the tube to just obtain a clear single
phase. Heat the tube in a 60-70 water bath for about 10 minutes, and then cool the
mixture to room temperature, and then in an ice bath. If no crystals have formed on cooling
in ice, add a few drops of water to the tube and scratch the inside wall of the tube under the
liquid surface with a glass rod. Collect the solid by suction filtration on filter paper in a
Hirsch funnel, wash the crystals with several mL cold water, and then dry the solid on filter
paper. Record MP.
b) Oxime Derivative Preparation
Disolve 0.2 gm. hydroxylamine hydrochloride in 2 mL. of water and 1 mL. of 3M NaOH. Add 0.2
gm. of your unknown to the above solution. Warm the mixture in a boiling water bath for 10
minutes. Cool in an ice bath and collect the crystals using vacuum filtration. Dry overnight and
determine the compounds melting range.

2. Derivatives for Aromatic amines, Phenols, Alcohols.

Procedure-- Add 2 mL of aqueous sodium hydroxide solution into a large test tube. Add the
amine, about 0.20 g if its a solid. Shake the solution vigorously and add about 5 drops of
benzoyl chloride. Stir vigorously for 10 minutes, and then few drops of aqueous HCl (this helps
the amide to crystallize). Cool on ice, filter the lumpy product through the Buchner funnel and
wash with 3x5 mL of cold water, then 2 x 3 mL of aq. HCl (to wash off unreacted amine), and
then 2 x 3 mL of NaOH/water (to wash off unreacted benzoyl chloride). Recrystallize using a

minimum of ethanol, perhaps adding water as necessary. Dry under suction and record MP. (the
filtrate may contain excess benzoyl chloride, decompose it by adding excess NaOH solution and
discard it after 15 min.)
3. Derivatives for Alcohols, Phenols.
3,5-Dinitrobenzoate Derivative Preparation
Dissolve 0.25 gm. Of sample in 0.25 g of 3,5-dinitrobenzoyl chloride and heat gently for few
minutes, cooled and poured into 10 ml of cold water taken in a beaker. The ppt. is filtered and
washed with cold water, dried, and subjected to MP determination.
4. Bromo derivatives for Phenols, Acids, Amines, Amino acids.
In a large test tube, add 0.2 mL of Bromine in 1 ml of glacial acetic acid. This solution is added to
a mixture of 0.2 gm of unknown dissolved in 1 mL glacial acetic acid and shaken vigorously.
Continue adding brominating solution dropwise, with mixing, until the orange bromine color
persists. Warm the solution on a water bath for few minutes. Add 2 mL of H2O and shake
vigorously. Collect the precipitate by filtration. Wash the precipitate with cold water.
Recrystallize from an appropriate Methanol: H2O mixture.
5. Derivatives for Acids.
a) Ethyl Ester derivative:
200 mg of compound is dissolved in 2 ml of ethanol, 2 drops of conc. H2SO4 is added and the
solution is heated to reflux for 5 min. The solution is allowed to stand for 10 min, 5 ml of
water is added and the solid is filtered.
6. Preparation of derivatives by Hydrolysis (for amides, anilides, esters etc.)
0.5 g of O.S. and 10 ml of 20% NaOH solution are boiled in a beaker for 15 min. and then cooled.
The mixture is then acidified with dil HCl. The solid is filtered, washed with cold water,
recrystallized from water of alcohol. Determine MP.
7. Derivatives for Aromatic Hydrocarbons. The main reactions carried out for the preparation of
derivatives for aromatic hydrocarbons are (a) nitration or (b) side chain oxidation.

(a) Nitration.
Nitro derivatives can be prepared for benzene, toluene etc. About 1 ml of fuming nitric acid and 1 ml of
conc. sulphuric acid are mixed and cooled in ice-water. About 0.25 ml of benzene or toluene (or other
hydrocarbon) is added to the nitrating mixture. Then the mixture is heated on a boiling water bath for
half an hour, till a drop of mixture poured into water crystallizes immediately. The mixture is then
poured into cold water taken in beaker and stirred well. The crystals are filtered at the pump,
recrystallised from dilute alcohol, dried and then melting point is noted.

(b) Side chain oxidation.

For aromatic hydrocarbons containing side chain like toluene or sidechain like xylenes, side chain
oxidation can be effected for the preparation of their derivatives. About 0.25 ml (0.25g) of the substance
is mixed with 0.25 g of sodium carbonate in 10 ml of water was taken in a 100 ml beaker and heated to
boiling (keep it on a wire gauze over a burner) and portionwise add 1 g of potassiumpermanganate
under gentle boiling until slight permanent pink colour is obtained. The excess pink colour of
permanganate is decomposed by addition of excess sodium bisulphate. The mixture is allowed to cool
and is carefully acidified with dilute sulphuric acid. The mixture is then again refluxed for 5 min and then
cooled. The excess of MnO2 is removed by addition of little sodium hydrogen sulphite, if required. The
white solid is filtered, recrystallised from warm water or alcohol, dried and MP was determined.