SCHEMATIC PROCEDURE FOR THE SEPARATION OF A

MIXTURE OF ORGANIC COMPOUNDS

PILOT SEPARATION:
To a small amount of the organic mixture taken in a DRY TEST TUBE, a little amount of ether
is added, shaken well and then allowed to stand
Residue Ether layer
Presence of ether It is taken in a separating funnel; sufficient amount of saturated solution of
insoluble sodium bicarbonate is added, shaken well and allowed to stand.(Repeated till
compounds. the acidic compound is removed completely)
Aqueous layer Ether layer
A small amount of Acidified with Sufficient amount of 10% of sodium hydroxide is added.
substance is 1:1 hydrochloric Shaken well and allowed to stand.
dissolved in acid. A solid
saturated separates out
sodiumbicarbonate Aqueous layer Ether layer
shows the
solution. Presence of Acidified with Sufficient amount of 1:1 hydrochloric
i) Dissolves with strong acidic 1:1 hydrochloric acid is added, shaken well and
brisk effervescence compounds like acid. allowed to stand
and regenerated on carboxylic acid A solid/turbidity Aqueous layer Ether layer
acidification using separates out It is neutralized Washed with
dil.HCl. Presence shows the with 10% water and the
of acidic Presence of sodium aqueous layer is
component. weak acidic hydroxide. An discarded. To the
ii) Dissolves compounds like oily liquid ether layer
without phenolic separates out anhydrous Na2SO4
effervescence and compound shows the or MgSO4 is added
no regeneration on
Presence of and decanted into
acidification.
basic a 100mL beaker,
Presence of
compounds followed by
neutral
like amines. evaporation of
components like
ether. Presence of
(i) Carbohydrates
neutral
(ii) Aliphatic
compounds like
diamide
Aldehyde,
(iii) Aromatic
Ketone,
amide
Ester,
Nitro benzene.

Pilot Report : The mixture contains ether soluble / insoluble ---------------- and ether soluble
-------------- components.

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BULK SEPARATION

1. Completely Soluble Mixture :

(a) Acid and Neutral Components : ( NaHCO3 Separation )

The given mixture is completely dissolved in ether and transferred into a separating
funnel. Ether solution is washed with aqueous solution of NaHCO3 and the process is
repeated for five or six times till the ether layer is free from acid. Every time the bottom
aqueous layer is collected separately from the ether layer. The aqueous layer is cooled and
acidified with conc. HCl. The regenerated acid is filtered at the pump, washed, dried and
recrystallized using hot water.
Ether layer is washed with water after ensuring that acid has been completely removed
from ether layer. It is then dried over anhydrous Na2SO4 and then ether is evaporated off to
collect the given neutral component.

(b) Acid and Base or Acid and Phenol : ( NaHCO3 Separation )

Same as before.

(c) Phenol and Amine or Phenol and Neutral Component: ( NaOH Separation )

The given mixture is completely dissolved in minimum quantity of ether and

transferred into a separating funnel. Ether layer is washed with 10 % NaOH solution five or
six times till the ether layer is free from phenol and collected the aqueous layer
separately. This aqueous layer on acidification with conc.HCl, regenerates the phenolic
component.
The ether layer is washed with water after ensuring that phenol has been
completely removed. It is then dried over anhydrous Na2SO4 or MgSO4 and then ether is
evaporated off to collect the other component. ( Basic or Neutral )

(d) Amine and Neutral Component : ( dilute HCl Separation )

The given mixture is dissolved in minimum quantity of ether and transferred into a
separating funnel. It is then washed with dilute HCl (5N) for five to six times till the entire
basic component is dissolved in dilute HCl. The aqueous layer is collected separately from
the ether layer. It is then cooled and made alkaline by the addition of NaOH solution to
regenerate the dissolved amine. The regenerated amine is collected separately and purified
further.The ether layer is washed with water after checking the complete removal of the basic
component from the mixture. It is then dried over anhydrous Na2SO4 and then ether is
evaporated off to collect the given neutral component.

2. Partly Soluble Mixture : ( Ether Separation )

This mixture can be separated based on the solubility of two components in ether.
The given mixture is treated with 20 to 30ml of ether and stirred. Ether insoluble component
remains in the bottom and ether soluble component remains as a top layer. It is filtered using
a filter paper to separate them. The residue is dried and then subjected to further purification.

2
 If ether layer contains acidic, phenolic or basic component, it can be separated respectively
by NaHCO3, NaOH or dil.HCl. (Refer before)
The soluble component present in ether is washed with water, dried over anhydrous
Na2SO4/MgSO4 and evaporated off ether to get ether soluble neutral component.

Systematic Analysis of an Organic Compound

Experiment Observation Inference

Nature of compound
1. Colour and Appearance Yellow solid or liquid May be Nitro compound
The colour and appearance Brown or dark coloured solid May be phenols or aromatic
of the given substance are or liquid amines
noted Colourless solid or liquid May be acid, amide,
carbohydrates,aldehyde,
ketone,ester etc.
2. Odour Fruity or pleasant odour Presence of ester, ketone etc
The odour of the given Phenolic odour Presence of phenol
substance is noted Bitter almond odour Presence of aldehyde
Fishy odour Presence of aromatic amine
3. Test for
Aromatic/Aliphatic Burns with a yellow sooty Aromatic compound.
i) Ignition Test : flame.
A small amount of the Burns with a yellow flame. Aliphatic compound.
substance is ignited on a
nickel spatula.
ii) Nitration Test :
A small of the substance is Yellow colour solution. Aromatic compound.
heated with a mixture of
1mL of Con.HNO3 &
Con.H2SO4, then it is cooled Colourless solution. Aliphatic compound.
and poured into a beaker
containing water.
4. Test for
Saturation/Unsaturation Decolourisation takes place Presence of unsaturated
i) Bromine water Test compound.
To a small amount of the
substance, 1mL of bromine Decolourisation takes place Presence of saturated and
water is added and shaken with the formation of white easily brominated
well precipitate. compounds like phenol,
amines.
.
No decolourisation takes Presence of saturated
place. compound.
ii) KMnO4 Test :
To a small amount of the Decolourisation takes place Presence of unsaturated
3
 substance dilute KMnO4 is with white precipitate compound.
added.
Decolourisation takes place Presence of saturated and
with the formation of black easily oxidisable compounds
precipitate. like phenol, amines and
aldehyde.

No decolourisation takes Presence of saturated

place. compound.

Lassaigne’s Test ( Nitrogen , Halogen , Sulphur )

Preparation of sodium fusion extract :

A small piece of sodium metal is taken in an ignition tube, heated and cooled. Then a
small amount of the organic compound is added. The mixture is heated strongly to red hot
condition, plunged into 5mL of distilled water taken in a mortar. The contents are ground, boiled,
cooled and filtered. This filtrate is the sodium fusion extract.

EXPERIMENT OBSERVATION INFERENCE

5. Test for Halogen
A few mL of silver nitrate is added White precipitate Presence of Chlorine.
to a small portion of the extract and
shaken well. Pale yellow precipitate Presence of Bromine.

6. Test for Sulphur

A few mL of a dilute solution of Violet coloured solution Presence of Sulphur.
sodium nitro prusside is added to a small
portion of the extract and shaken well.

7. Test for Nitrogen

A few mL of freshly prepared Blue or Green solution Presence of Nitrogen.
ferrous sulphate solution is added to
the extract, boiled, cooled and
acidified with dilute H2SO4.

Preliminary test:

8. Sodium bicarbonate test The compound dissolves Presence of carboxylic acid.

To a small amount of the compound with brisk effervescence
a saturated solution of sodium and evolution of CO2.
bicarbonate is added.
9. Sodium hydroxide test

To a small amount of the compound,

4
10% solution of sodium hydroxide Soluble in cold Presence of acid or phenol
is added. The solution is heated
strongly. Oily drops are obtained. Presence of anilide.

Substance dissolves on Presence of acid, esters or

heating and a white amide.
precipitate is obtained on
acidification with
Con. HCl.

10. Conc. Sulphuric acid Test

To a small amount of the compound,
1mL of Con.H2SO4 is added and Charring or blackening Presence of Carbohydrate.
warmed takes place with
effervescence

11. Neutral Ferric Chloride Test

A small amount of the substance is
shaken well with a few drops of A Violet or green solution Presence of phenol or
neutral ferric chloride solution is observed phenolic derivatives.

12. Borsche’s Reagent Test

To a small amount of the substance
1mL of Borsche’s reagent, a drop of A yellow or red orange Presence of aldehyde or
Con. HCl are added, heated on a precipitate is obtained ketone.
water bath and cooled.

13.Tollen’s Reagent Test

To a small amount of the organic
compound, a few mL of Tollen’s A bright silver mirror or a Presence of aldehydes or
reagents A and B are added. The black precipitate of silver reducing sugars.
mixture is warmed on a boiling is obtained
water bath .

14. Fehling’s Reagent Test

To a small amount of the organic
compound, a few mL of Fehling’s A red-brown precipitate is Presence of reducing sugars
reagents A and B are added. The obtained. or aliphatic aldehydes.
mixture is heated on a boiling water
bath.

15.Mulliken and Barker Test

To a small amount of substance

5
1mL of freshly prepared ferrous Reddish brown precipitate Presence of nitro compound
ammonium sulphate solution is of Iron (III) hydroxide is
added, followed by 1 mL of 2N observed
alcoholic KOH reagent and shaken
well.

Confirmatory Test

1.Carbohydrates

i) Molisch Reagent Test

A small amount of the substance is
dissolved in water and a few drops A violet or red ring is Presence of carbohydrate is
of Molish reagent (saturated solution formed at the junction of confirmed.
of α-naphthol in alcohol) is added two layers.
and shaken well.About 1mL of
conc.H2SO4 is added carefully along
the sided of the test tube.

ii) Seliwanoff’s test

To a small amount of the substance
few drops of Seliwanoff’s reagent (a A cherry-red condensation Presence of ketoses
solution of resorcinol and HCl) is product is obtained within
added. The solution is then heated 2 min.
on a water bath for two minutes.
Blue-green condensation
product is obtained after 2 Presence of aldoses.
min or no cherry-red is
2. Carboxylic acid observed.

i) Esterification Test
To a small amount of the compound,
2 mL of alcohol and 0.5 mL of A pleasant fruity smell of Presence of carboxylic acid
con.H2SO4 are added and heated ester is observed
strongly. The hot mixture is cooled
and poured into 10 mL of aq.
Sodium hydroxide.

ii) Fluorescein Test

A small amount of the organic
compound is mixed with an equal An intense greenish Presence of dicarboxylic
amount of resorcinol and 1 mL of yellow fluorescence is acid is confirmed.
conc. H2SO4.The mixture is heated. observed
The hot solution is then poured into
10 mL of water, then add excess of No intense greenish Presence of Monocarboxylic
NaOH solution. yellow fluorescence is acid is confirmed

6
:- observed
3. Phenol

i) Liebermann’s Test
A small amount of the substance is
heated with crystals of sodium Presence of phenol is
nitrite (NaNO2) in a dry test tube and Red solution is obtained confirmed.
cooled. Two drops of conc. which turns blue or green
H2SO4 are added and shaken well. A on adding sodium
portion of the mixture is poured into hydroxide solution.
distilled water.

ii) Phthalein fusion Test

A small amount of the organic
compound is mixed with an equal Presence of monohydric
amount of phthalic anhydride and A pink or red colour is phenol.
heated with 2 mL of conc. H2SO4. formed.
The mixture is heated strongly and
poured into 20 mL of water and
excess NaOH solution is added.

iii) Azo dye test

About 1 mL of aniline is dissolved
in dilute HCl, and it is mixed with Presence of phenolic
saturated solution of sodium nitrite A scarlet red or orange red compound is confirmed.
constant shaking under ice cold dye is obtained
condition. To the diazotized
solution, 1 mL of the phenolic
compound dissolved in 10% NaOH
solution is added and shaken well.

4. Amines

To the compound dissolved in dilute

HCl added sodium nitrite (NaNO2) Presence of primary amine
under ice cold condition. Pale yellow oily liquid is
formed
Presence of secondary amine
Yellow oily liquid is
formed

Presence of tertiary amine

Green solution is obtained.
Test for primary amine:

To the above mixture, an ice cold

7
solution of β - naphthol in NaOH is A Scarlet red dye is Presence of primary amine is
added and shaken. obtained confirmed.

Test for secondary amine:

Yellow oil formed is extracted in
ether and evaporated off ether. Then A Green or blue solution is Presence of secondary amine
added few drops of phenol and con. obtained is confirmed.
H2SO4. Heated and poured into
water containing NaOH.

Test for tertiary amine:

To the substance potassium

ferricyanide is added and shaken. A Green colour solution is Presence of tertiary amine is
obtained. confirmed.

5. Amide

i) Sodium hydroxide test

A small amount of the compound is
boiled with 10ml of 10% NaOH A white precipitate is Presence of aromatic amide
solution. After ammonia gas is obtained.
evolved, the solution is cooled and
acidified with con.HCl. No white precipitate Presence of aliphatic amide

ii) Biuret Test

A small amount of the compound is
heated strongly until there is no AViolet coloured solution Presence of diamide is
evolution of NH3. The sublimate is obtained. confirmed.
formed is scrapped and dissolved in
water. A few drops of dilute CuSO4
solution is added followed by a few
ml of aq. NaOH.

6. Aldehyde

i) Schiff’s Test
A few ml of Schiff’s reagent is
added to a small amount of the A Pink coloured solution is Presence of aldehyde is
compound and shaken well. formed. confirmed.

ii) Schiff’s base formation

To a few mL of aniline, the liquid

8
substance is added and shaken well. A yellow colour is Presence of aromatic
observed aldehyde is confirmed
7. Ketone

i) Legal’s Test

A small amount of the organic

compound is added to 1 mL of A wine red colour is Presence of Methyl ketone
freshly prepared dilute sodium nitro formed with NaOH. (CH3-C=O) is confirmed.
prusside solution, followed by 1 mL On adding acetic acid, a
of NaOH solution. purple or blue colour is
formed.

No blue colour is seen. Presence of aromatic ketone

is confirmed.
ii) A small amount of the organic
compound is dissolved in alcohol Red solution turning blue Presence of ketone is
and is treated with m - dinitro on adding CH3COOH. confirmed.
benzene and NaOH solution.

8. Anilide

Azo dye Test .

A small amount of the organic
compound is heated with sodium A Scarlet red dye is Presence of anilide is
hydroxide . Azo dye test is formed. confirmed.
performed with the oily drops
formed.

9. Nitro compound:

i) Mulliken and Barker Test

A small amount of the organic
compound is dissolved in alcohol.
Zn dust and solid NH4Cl are added
and the mixture is boiled, cooled,
filtered and divided into two parts.

(a) To one part of the filtrate added

Fehling’s reagents A and B and A Reddish brown Presence of nitro compound
heated in a water bath. precipitate is obtained is confirmed.

(b) To the second part of the filtrate

a few ml of Tollen’s reagents A and A Black precipitate is Presence of nitro compound
B are added and warmed. obtained. is confirmed.

9
ii) Dye test
A small amount of the substance is
mixed with a few granules of tin and .A Scarlet red dye is Presence of nitro compound
2 mL of conc. HCl heated and obtained is confirmed.
filtered. Azo dye test is performed
with the filtrate.

10. Ester

i) Sodium hydroxide test

A small amount of the compound is
heated with 10% NaOH solution, A white precipitate is Presence of sster is
cooled and acidified with conc.HCl. formed. confirmed.

ii) Hydroxamic acid Test

A few drops of the substance, solid
hydroxylamine hydrochloride and a
few ml of 10% NaOH solution are A violet colour is formed. Presence of ester is
heated strongly, cooled, acidified confirmed.
with conc. HCl. A few drops of
FeCl3 solution is added and shaken
well.

PREPARATION OF DERIVATIVE

1. Anilide : Bromo derivative

About 1 g of the organic compound is dissolved in 5 ml of glacial acetic acid. About 2ml of
Br2 in acetic acid is added to the above solution. The mixture is shaken vigorously for 10 minutes
and poured into 50 ml of ice cold water. White crystals separated out are filtered and dried.

2. Carbohydrate : Osazone derivative

About 1 g of the organic compound is dissolved in 3 ml of water. About 2 g of phenyl
hydrazine hydrochloride and 2 g of CH3COONa are added to the above solution. Few drops of
acetic acid are added and the mixture is heated on a water bath for about 10 minutes and cooled.
Yellow spongy crystals separate out.

3. Diamide : Oxalate derivative

About 1 g of the organic compound is shaken with a saturated solution of oxalic acid. White
solid separates out.

4. Monocarboxylic acid : Amide derivative

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 0.5g of organic compound is treated with PCl5 in a dry beaker, stirred till a clear liquid is
obtained. This clear liquid is added to 3 ml of liquor NH3. Solid formed is filtered, dried and
recrystallised with methanol.

5. Dicarboxylic acid : Anhydride derivative

About 1 g of the organic compound is taken in a china dish and covered with an inverted
funnel. It is then heated over a wire gauze with a low flame until needle shaped crystals are
obtained on the cooler part of the funnel.

6. Aldehyde and Ketone : Semicarbazone derivative

About 1 g of the organic compound is dissolved in alcohol. 10 ml of a saturated solution of
semicarbazide hydrochloride containing sodium acetate. The mixture is heated for about 10
minutes and then cooled. White crystals are obtained.

7. Phenol : Aryloxy derivative

2 ml of phenol is taken in a R.B. flask fitted with a refulx condenser. Then added
3 g of chloro acetic acid and NaOH solution till the solution is basic. The mixture is refluxed for
one hour, cooled and poured into ice. Neutralised with con.HCl. White precipitate formed is
filtered, dried and recrystallised with water.

7. 1 Phenol : Benzoyl derivative

About 5 g of the organic compound is dissolved in 15 ml of 10% NaOH solution. About
2 ml of benzoyl chloride is added. The mixture is shaken vigorously for about 15 minutes.
Colourless solid separates out.

8. Primary Amine : Acetyl derivative

2 ml of amine is mixed with 5 ml of glacial acetic acid and 5 ml of acetic anhydride. The
mixture is refluxed for one hour. Cooled and poured into ice. The derivative formed is filtered,
dried and recrystallised with dilute alcohol.

9. Sec. and Tert. Amines : Picrate derivative

1 g of picric acid is dissolved in benzene and to this solution added 1 ml of amine and
shaken well. The derivative formed is filtered is filtered and dried.

10. Solid phenols and Amines : Benzoyl derivative

About 5 g of the organic compound is dissolved in 15 ml of 10% NaOH solution. About
2 ml of benzoyl chloride is added. The mixture is shaken vigorously for about 15 minutes.
Colourless solid separates out.

11. Nitro compound : Polynitro derivative

About 5 ml of the organic compound is taken in a R.B. flask. Nitrating mixture (6 ml of
con.H2SO4 and 5 ml of con.HNO3) is prepared separately. The nitrating mixture is added in drops
to the organic compound and refluxed for 15 minutes . The mixture is cooled and then poured into
ice . Yellow crystals separate out.

11
12. Ester and Mono amide : Acid derivative
About 1 g of the compound is mixed with 15ml of 10% NaOH solution and the mixture
is refluxed for about 10 minutes and cooled. Con.HCl is added in drops with constant stirring till it
becomes acidic. White solid separates out

Report:
I. Nature of the compound
a) Aliphatic / Aromatic
b) Saturated / unsaturated

II. Presence or absence of characteristic elements

a) Nitrogen :
b) Chlorine :
c) Bromine :
d) Sulphur :

III. Functional group present :

PREPARATION OF BENZHYDROL FROM BENZOPHENONE

AIM: To prepare benzhydrol from benzophenone

12
CHEMICALS REQUIRED:

Benzophenone-2 g
Sodium hydroxide pellets-3 g
Zinc dust-3 g
Rectified spirit-15 mL (15-1)
REACTION:

When benzophenone is reduced by zinc dust in presence of NaOH, benzhydrol is

formed.

2[H] 2[H]

Zn / −OH

PROCEDURE:

2 g of benzophenone, 3 g or NaOH and 3g of Zn dust and 15 mL of rectified spirit are

taken in a R.B. flask fitted with a refluxing condenser. The reaction mixture is refluxed carefully
for an hour by heating R. B. flask over the copper water bath. After the reaction is over the
reaction mixture is cooled to 60ºC and filtered through suction rapidly to remove Zn dust from
the reaction mixture.

During filtration, care should be taken to keep Zn dust always in solution. The filtrate is
poured into ice, followed by the addition of HCl. The precipitate is filtered, dried and
recrystallized from alcohol.

RESULT:

The yield of benzhydrol is about --------- %

PREPARATION OF ANTHRAQUINONE FROM ANTHRACENE

AIM: To prepare anthraquinone from anthracene

CHEMICALS REQUIRED:

13
 Anthracene -2.5 g
Acetic acid - 30 mL
Chromic oxide -5 g

REACTION:

Anthracene when oxidized by CrO3, anthraquinone is formed.

CH3COOH CH3COOH

PROCEDURE:

2.5 g of anthracene is dissolved in 15 mL of acetic acid in a R.B flask fitted with a

reflux condenser. When anthracene is dissolved, the solution containing 5 g of Chromic oxide in
10 mL of water is added slowly to anthracene. When all CrO3 is added, the solution is refluxed
for 30 min and then poured into ice. The precipitate formed is purified and dried. Small amount
of the anthraquinone formed is purified by sublimation.

RESULT:

The yield of anthraquinone is about --------- %

PREPARATION OF m-NITROANILINE FROM m-DINITRO BENZENE

AIM: To prepare m-nitroaniline form m-dinitrobenzene.

CHEMICALS REQUIRED:

14
 m-dinitrobenzene
Rectified spirit
Ammonium sulphide

REACTION:

m-dinitrobenzene undergoes partial reduction in the presence of ammonium

sulphide to give m-nitroaniline.

(NH4) 2S

PROCEDURE:

The m-dinitrobenzene is taken in a R. B. flask and it is dissolved in 10 ml of rectified

spirit, followed by the addition of 15 mL of ammonium sulphide solution. It is then refluxed for
about 30-45 minutes. Then the reaction mixture is poured into ice. The product formed is filtered
and dried. A small portion of this is recrystallized with dilute alcohol.

RESULT:

The yield of m-nitroaniline is about ---------- %

PREPARATION OF m-NITROBENZOIC ACID FROM METHYLBENZOATE

AIM: To prepare m-nitrobenzoic acid from methyl benzoate

CHEMICALS REQUIRED:

15
 Methyl benzoate-5 mL
Conc. H2SO4-7.5 mL
Fuming HNO3- 5mL
Sodium hydroxide (10%)-20 mL

REACTION:

Methyl benzoate on nitration with nitrating mixture yields m-

nitromethylbenzoate which on hydrolysis with sodium hydroxide reacts to give m-
nitrobenzoic acid.

NaOH / Δ

H+

m-nitrobenzoic acid

PROCEDURE:

(a) m-nitrobenzoate

About 7.5 mL of Conc.H2SO4 and 5 mL of fuming HNO3 are mixed in a R. B. flask

under cooling. 5 mL of methyl benzoate is added in drops to the nitration mixture with cooling.
When the addition is over the reaction mixture is shaken well for 30 minutes at room
temperature and poured into ice. m-nitrobenzoate formed is filtered and dried.

(b) m-nitrobenzoic acid

m-nitrobenzoate and 20 mL of 10% NaOH are taken in a R. B. flask fitted with a

reflux condenser and the reaction mixture is refluxed for 45 minutes. Then it is poured into water
and acidified with dil. HCl. The acid formed is filtered and dried. A portion of it is recrystallised
using water.

RESULT:

The yield of m-nitrobenzoic acid is ---------- %

PREPARATION OF BENZANILIDE

AIM: To prepare benzanilide from benzophenone

CHEMICALS REQUIRED:

Benzophenone- 2 g

16
 Rectified spirit- 5 mL
Hydroxylamine hydrochloride- 1.5 g
Sodium hydroxide- 2 g
Conc. HCl - 1mL

REACTION:

When benzaldehyde is condensed with hydroxylamine, one water molecule is removed

and gives benzophenoxime or benzophenone oxime. This undergoes Beckmann rearrangement
followed by hydrolysis gives benzanilide.

Conc.HCl

Dry ether

PROCEDURE:

(a) Benzophenone oxime

2 g of benzophenone is placed in a R. B. flask. To this 5 mL of rectified spirit, 1.5 g of

hydroxylamine hydrochloride, 2 g of sodium hydroxide and 15 mL of water are added. The
contents of the flask are cooled with rise in temperature. Reaction mixture is refluxed using a
water condenser on a wire gauze for 10 minutes. The contents are cooled by pouring into a
beaker containing ice and neutralized with conc. HCl. The precipitated oxime is filtered washed
with cold water, dried under suction.

(b) Benzanilide (Beckmann rearrangement)

2 g of the above prepared benzophenone oxime is placed in a dry iodine flask and
dissolved in dry ether. To this solution of oxime 1 mL of conc. HCl is added in portionwise. The
contents are shaken for 10 minutes, ether layer is evaporated off and to the residue is added cold
water to precipitate out benzanilide. The precipitated formed is filtered, washed with water and
dried under suction. A part of benzanilide is recrystallized from aqueous alcohol.

RESULT:

The yield of benzanilide is --------- %

PREPARATION OF SYM-TRIBROMOBENZENE

AIM: To prepare sym-tribromobenzene from aniline

CHEMICALS REQUIRED:
Aniline - 7 mL
Glacial acetic acid - 2 mL
Br2 / CH3COOH

17
 Ethanol (15 mL) + Benzene (7 mL)
H2SO4 – 1 mL
NaNO2 – 2.5 -3 g

REACTION:

Aniline reacts with Br2 / CH3COOH giving 2,4,6-tribromoaniline. This undergoes

deamination on further reaction with sodium nitrite to give sym-tribromobenzene.

NaNO2 / H2SO4
Br2 / CH3COOH
Δ

PROCEDURE:

(a) 2,4,6-tribromoaniline

7 mL of aniline is placed in Iodine flask and dissolved in about 2 mL of glacial acetic

acid. To this solution, is added a solution of Br2 / CH3COOH in portionwise with shaking. The
addition is continued until the contents of the flask remains permanent pale yellow colour. The
contents are shaken for 10 minutes and poured into cold water. The precipitate 2,4,6-
tribromoaniline is filtered, washed with water to make the precipitate free from excess Br2 /
CH3COOH.The precipitate is dried under vacuum. The yield is recorded.

(b) sym-tribromobenzene
4 g of 2,4,6-tribromobenzene is taken in a R. B. flask, to this, a mixture containing
ethanol (15 mL ) and benzene (7mL) is added. To the contents of the flask, 1mL of conc.H2SO4
is added and refluxed in a water bath using a condenser. About 2.5- 3 g of NaNO2 is added in
one lot through the top of the condenser, and the mixture is shaken.The reaction mixture is
transferred into a dry beaker, allowed to cool. The sym-tribromobenzene that separates out is
filtered off under suction, washed thoroughly with water and dried. A small amount of the
sample is recrystallised from hot rectified spirit.

RESULT:

The yield of sym-tribromobenzene is ---------- %

Thin Layer Chromatography
AIM
To calculate the Rf value of o-nitrophenol and p-nitrophenol by TLC

PRINCIPLE:

It is an adsorption chromatography. The sample is applied at the base line on the plate coated
with adsorbent, and then it is dipped in the chamber containing solvent which acts as mobile
phase. The solvent rises along the stationary phase due to capillary action.

18
 The different component in the sample are adsorbed to different extent on the adsorbent
and when solvents starts raising the different components move upward at different rates and this
leads to separation of components. The more polar the molecule, the shorter the distance it will
travel during TLC development.

Procedure:
a)TLC Plates: A commercially available TLC plate was cut into a rectangular shape of a
dimension of 6cm x 2cm and this was used for the experiment
b) Stationary phase: Silica gel coated on aluminium sheet was considered as stationary phase.
c) Mobile phase: A 20% ethylacetate in hexane was used as mobile phase. It was prepared by
mixing 2mL of ethyl acetate and 8mL of hexane.
d) Preparation of sample solution: The given samples were dissolved in ethyl acetate. The
sample solutions were applied approximately 1cm above the lower edge of TLC plates by means
of capillary tube. The spots were dried and the plates were developed in TLC chambers by the
ascending technique. Before chromatography, the chamber containing the mobile phase was
covered with a lid for approximately 10 minutes to enable the pre-saturation with mobile phase
vapour. After development, the plate was withdrawn from the chamber and dried out at room
temperature. All the components were then detected by exposing the plates to iodine vapour in a
closed chamber by approximately 2 minutes. The components were detected as yellow spots.
The Rf value of the components were calculated

Distance moved by the component

Rf =

Distance moved by the solvent

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O – ortho nitrophenol
P – para nitrophenol
M – Mixture of ortho and para nitrophenol
Rf (O) = x1 / y
Rf ( P) = x2 / y

Result:
a) The Rf value of o-nitrophenol =
b) The Rf value of p-nitrophenol =

20
 Extraction of Lactose from Milk
Chemicals required:
Milk-500ml
Dil. Acetic acid-100 ml
Calcium carbonate- 2g
Rectified spirit – 50 ml

Procedure

About 500ml of milk is diluted with water and centrifuged to remove the fat. The fat
appears on the top of the centrifuging tube is removed. The clear thin milk obtained after
centrifugation is called skim milk. To the clear skim milk about 50 ml of dilute solution of acetic
acid is added slowly with constant stirring. A white solid separates out called casein.The
precipitated casein is filtered off and the filtrate is neutralized with dilute aqueous sodium
hydroxide solution. The solution is made very slightly acidic by adding 2 or 3 drops of acetic
acid to the neutralized solution. The solution is boiled on wire gauze with 2 g of calcium
carbonated solid. The mixture is stirred well and filtered hot. The solution is concentrated by
boiling and converted into one third volume. About 3 volumes of rectified spirit is added and the
solution is filtered. White crystals of lactose separate out. The white solide is filtered off at the
pump, washed with alcohol and dried in air.

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 Extraction of Citric acid from Lemon

Chemicals required:

Lemon juice-100ml
Calcium chloride (10%) – 2 ml
Dilute ammonia – 15 ml
Dilute sulphuric acid- 10ml

Procedure

About 100mL of lemon juice is taken and diluted with double the amount of water. The
juice is neutralized by adding dilute ammonia slowly with constant stirring. The citric acid
present in the juice is now converted into ammonium citrate. To the solution of ammonium
citrate, about 2 ml of a 10% aqueous solution of calcium chloride is added. The resulting solution
is heated to boiling and set aside for 30 minutes. A white precipitate of tricalcium citrate is
formed. The precipitate is washed well with water and the washing is decanted. From the
tricalcium citrate, citric acid can be obtained by treating with dilute sulphuric acid. The white
precipitate of calcium sulphate formed is filtered and the filtrate is concentrated to obtain crystals
of citric acid.

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