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ESTIMATION OF PHENOL

AIM:

To estimate the amount of phenol present in the whole of given solution

PRINCIPLE:

The estimation Is based on the following reaction

However a standard solution of bromine is not used for this purpose. Instead a
mixture of bromate bromide (wrinklers solution) which readily liberates bromine in the
presence of acid is used. Also the strength of the thio sulphate solution does not vary with
this and this is fairly stable

The excess of bromine is treated with KI, so that the liberated iodine could be
treated against sodium thiosulphate, which inturn is standardised by potassium
dichromate solution.

PROCEDURE:

STANDARDISATION OF SODIUM THIOSULPHATE SOLUTION:

(i) Prepare a standard solution of potassium dichromate

(ii) Pipette out 20ml of syd. Potassium dichromate solution in a clean conical
flask add 5ml of conc. HCL and 40ml of 10% KI and titrate the liberated
iodine against sodium thiosulphate in burette.
(iii) When the solution becomes pal yellow add one drop of starch indicator and
continue the titration
(iv) The end point is the appearance of green colour.
(v) Repeat the titration fod concordant value
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ESTIMATION OF WRINKLERS SOLUTION:

(I) Conduct a similar experiment to standardise the bromate bromide mixture

(II) Pipette out 20ml of this solution into a clean conical flask
(III) Add 5ml of conc. HCL and 10ml of 10%KI and titrate the liberated iodine
against sodium thiosulphate solution using starch indicator.
(IV) End point is the disappearance of colour
(V) Repeat the titration for concordant value

ESTIMATION OF PHENOL:

(i) Make up the given phenol solution into 100ml in a std. measuring flask
(ii) Pipette out 20ml of the solution into stoppered conical flask
(iii) Dilute it by adding 50 ml of water
(iv) Add 40ml of wrinklers solution from burette slowly with constant shaking
(v) Then add 5ml of conc. HCL 10ml of 10% KI solution
(vi) Titrate the liberated iodine against thiosulphate using starch indicator
(vii) End point is the disappearance of colour
(viii) Repeat the titration for concordant value

RESULT :

Amount of phenol present in the given solution =

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STD DICHROMATE Vs STD THIOSULPHATE

S.No Vol of sodium Concordant

Vol of dichromate Burette reading thiosulphate value
solution (ml) solution
V1(ml) V2 (ml)
Initial Final

Strength of K2Cr2O7 (N1) =

Volume of dichromate solution (V1) =

Volume of sodium thiosulphate solution (V2) =

Strength of thiosulphate solution (N2) =

According to law of volumetris analysis,

V1*N1 = V2*N2

ESTIMATION OF WRINKLERS SOLUTION:

S.No Vol of Burette reading Vol of Concordant

Wrinklers Initial Final Thiosulphate Value
Solution (ml) (ml) Solution
V1 (ml) V2 (ml)
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ESTIMATION OF PHENOL:

S.No Vol of Burette reading Vol of Concordant

phenol Initial Final Thiosulphate Value
Solution (ml) (ml) Solution
V1 (ml) V2 (ml)
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PREPARATION OF M- DINITROBENZENE

AIM:

To prepare the solution of M-Dinitrobenzene

PRINCIPLE:

M-Dinitrobenzene is prepared by nitraton of nitrobenzene

Nitration is effected by fuminf nitric acid in the presence of sulphuric acid

REQUIREMENTS:

Nitrobenzene
Fuming nitric acid
Conc. Sulphuric acid
PROCEDURE:

7ml of fuming nitric acid is taken in a round bottomed flask and 10ml of conc.
H2So4 is added At the time of cooling a little of 5ml nitrobenzene is added in small
quantities to the flask and shake well after each addition Finally it is heated for about 45
mins by immersing in a boiling water bath. Till a small quantities of mixture when added
to small quantities of water in a test tube gives solid immediately. The contents of the
flask are then poured into a fine stream while still hot into 100ml water contained in a
beaker. The mixture is shaken very vigorously
During addition, M-nitrobenzene separates as a solid which is filtered at the pump. Wash
several times with water, dried and yield is noted.

RESULT:
Amount of M-Nitrobenzene obtained =
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PREPARATION OF ACETANILIDE

AIM:

To prepare a solution of acetanilide

PRINCIPLE:

Aniline is refluxed with glacial acetic acid in the presence of fused sodium acetate
where acetanilide is obtained

REQUIREMENTS:

Freshly distilled aniline

Glacial acetic acid
Freshly fused sodium acetate
PROCEDURE:
5ml of aniline, 6ml of glacial acetic acid and 2gm of freshky fused sodium acetate
is taken in a round bottomed flask.
The mouth of the flask is fitted with a long air condenser and gently refluxed by
heating on wire gauze. Heating is continued for 2-3 hrs.
The mixture is poured white hot into100ml cold water contained in a beaker with
vigorous strring. It is cooled and filtered at the pump.
The slightly coloured substance is dissolved in about 150ml boiling water.
Heated with 1gm of animal charcoal and filtered through hot water funnel. The
precipitated acetanilide is filtered and washed with water and dried. Yield is noted

RESULT:

Amount of acetanilide obtained is =

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PREPARATION OF BENZOIC ACID FROM ETHYL BENZENE

AIM:

To prepare benzoic acid from ethyl benzene

PRINCIPLE:

Ethyl benzene is hydrolysed to sodium salt of benzoic acid by a solution of sodium

hydroxide

Benzoic acid is obtained from sodium benzoate from acetification

REQUIREMENT:
Ethyl benzene
Sodium hydroxide
PROCEDURE:
2gm of NaOH is dissolved in about 20ml of water taken in round bottomed flask.
2.5gm of benzoate is added to round bottom flask. A few porous pieces are added to
round bottom flask. The flask is then fitted with a zie bag condensor and heated over wire
gauze for 45 mins.The hydrolysis is complete when no more oily drops are seen in the
flask. Contents are now cool and transform completely to a beaker. Conc. HCL is added
with constant strring till the solution is diatinctly agitated. The precipitated benzoic acid
is filtered, washed and dried. The yield is noted.
RESULT:
The yield of benzoic acid is =
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PREPARATION OF BENZOIC ACID FROM BENZALDEHYDE

AIM:

To prepare benzoic acid from benzaldehyde

PRINCIPLE:

When benzaldehyde is oxidised using alkaline permanganate solution of benzoic

acid is form acid sodium salt.the precipitated manganese dioxide is disoolved using
acidified soudium sphone solution or acidification benzoic acid precipitates

REQUIREMENTS

Benzaldehyde 5 ml

Kmno4 6 gms in 100ml of H2O

Sodium benzoate 2.5 gms in 20ml of H2O

PROCEDURE

The benzaldehyde is mixed with aqueous solution of sodium carbonate taken in

250 ml round bottomed flask fitted with reflux condenser.the contents are heated slowly
adding Kmno4 solution in the flask from the top of the condenser until the solution is
pink then the contents are refluxed with 90 mins the flask is cooled and transformed in
250 ml beaker aqueous saturated solution of sodium bisulphate is added with constant
stirring with aprecipitated manganese dioxide gets disoolved to liberate free acid
concenrtrated Hcl is added till the precipitation is completed filtered off washed with
water and dried yield is noted.

RESULT:

Yield of benzoic acid=

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HYDROLYSIS OF METHYL SALICYLATE TO SALYCYLIC ACID

AIM:

To estimate the salycylic acid by hydrolusis of methyl salicllate

PRINCIPLE:

Esters can be hydrolysed into their carboxylic acids either in acidic or basic
medium under refluxing condition. In this requirement, methyl salicylate, an ester is
refluxed with aqueous base. Due to the presence of base, the carboxylic acid is ionized
and gives sodium salicylate. The reaction mixture is subsequently acidified using
sulphuric acid, which converts the anion into the fully protonated acid i.e., salicylic acid.
The salucylic caid formed is filtered on a suction pump and purifird by crystallization.

REQUIREMENTS:

CHEMICALS APPARATUS
Methyl salicylate- 4ml R.B.Flask 250ml
NaOH solution(10%):25ml Reflux condenser
3M H2SO4 cl : 50ml glass rod
Litmus paper(blue) Wire guaze, funnel

PROCEDURE:

Take sodium hydroxide solution in a100ml round bottomed flask. To the NaOH
solution add 4.2gm(0.03mol) of methyl salicylate (NOTE: A white solid is formed, but it
will dissolve when the mixture is heated). Add 2 pieces of porcelain to prevent bumping
of the reaction. Place a reflux condenser, over the flask and reflux it for 20mins using
heating mantle. After the 20mins, transfer the reaction mixture to a beaker, cool it by
placingthe beaker in water bath and carefully add enough sulphuric acidto make solution
acidic. Filter the formed salicylic acid by vaccum filteration. Dry the product.

RESULT:

Yield of salicylic acid is =

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CONVERSION OF ACETONE TO IODOFORM

AIM:

To prepare iodoform from acetone by substitution.

PRINCIPLE:

Iodoform can be prepared from acetone by substitition with alkali iodine solution
as per the reaction

PROCEDURE:

Dissolve 0.1gm or 2ml of acetone in a 5-8ml of distilled water to get a

homogenous solution. Add 2ml of 5% NaOH solution and potassium iodide and Iodine
reagent (prepared by dissolving 20gm of KI and 10gm of I2 in 100ml of distilled water)
dropwise with shaking until a definiet dark of iodine persists. Allow to tand for 5mins. If
no yellow precipitate is formed, warm in a beaker of water at 60C add more drops of
iodine reagent if the faint iodine color disappears. Continue the addition of the reagent
until the rark color is not discharged after 2mins heating at 60C . remove the excess of
iodine by the additio of few drops of NaOH solution with shaking. Dilute with equal
volume of water and allow to stand for 10-15mins. Filter off the yelloe precipitate, dry
upon pads of filter paper and weigh till steady reading.

RESULT:

The yield of iodoform is

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ANALYSIS OF PROTEINS

S.No EXPERIMENT OBSERVATION INFERENCE

1 Biuret test: Substance in strong Bluish violet/pink Presence of proteins
NaOH solution + few drops of Dil. colour
CuSO4 solution
2 Xanthoprotein test: Warm the given Yellow colour turns Preaence of proteins
sample with HNO3 to orange on
addition of NaOH
solution
3 Millons test: solution of protien + White precipitate Presence of protein
millons reagent(prepard by the turns to red on with OH group
dissolution of mercurous and heating
mercuric nitrate in HNO3)
4 Molischs test to about 0.1gm of Violet ring at the Presence of protein
substance in 1ml ater add 1ml of junction of two with carbohydrate as
alcoholic -napthol solution, shake layers prosthetic group
well and add conc.H2SO4
dropwise through the side of the
test tube
5 Lead sulphide test: Boil the protein Black precipitate Presence of protein
solution with NaOH and lead with thiol/disilphide
acetata solution linkages
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ANALYSIS OF ORGANIC COMPOUND

S.No Experiment Observation Inference

1 Colour Yellow Presence of
Colour of the substance is noted aromatic nitro
compound,
Dark brown diketones
Presence of
Colourless aromatic amine,
phenol
Presence of
carboxylic acid,
aldehyde, ketone,
ester, carbohydrate
2 Smell Pleasant/Fruity Presence of ester
Odour of the substance is noted Phenolic Presence of Phenol,
Napthol
Pungent Presence of
aromatic compound
Aniline-like Presence of
aromatic amine
3 Solubility:
TO about 0.1g of substance Soluble in cold water Presence of
12ml of distilled water is added carbohydrate,
and shaken well polyhydric phenol,
certain acids
Soluble in hot water May be carboxyliv
acid or phenol
4 Litmus test:
To about 0.1gm of the Blue turns red May be carboxylic
subsatnce added 2ml water and acid or phenol
shaken well. Tested with the Red turns blue May be amine
litmus paper
5. DETECTION OF ELEMENTS :
Preparation of Sodium fusion extract
A small dru pellet of sodium is melted in an ignition tube. About 0.1 of the
substance is added, heated gently at first and then strongly till the botton of the tube
becomes red hot. The red hot end of the tube is plunged into 10ml of distilled water in a
china dish, ground well, boiled and filtered. The filtrate/extract is used for the following
tests.
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5a To about 1ml of the fusion Blue of green Presence of

extract, added a few crsytals of precipitate Nitrogen
ferrous sulphate, boiled and
cooled acdified with 2ml of
dil.H2SO4
5b To about 1ml of the extract, White precipitate Presence of chlorine
1ml of dil.HNO3 is added, soluble in aq.NH3
cooled, and then added silver Pale yellow
nitrate solution Presence of
precipitate partially
soluble in aq. NH3
Deep yellow Presence of iodine
precipitate insoluble
in aq. NH3

5c To about 1ml of the extract, few Violet colour Presence of sulphur

droops of sodium noyroprusside
solution is added
6. Test for Aliphatic or Aromatic
A Ignition test: Burns with a Presence of
About 0.2g of the substance is luminous smoky aromatic compound
ignited in a nickel spatula flame Presence of
Burns with a non- aliphatic compound
luminous flame
B Nitration test
About 0.2g of the substance is Yellow Presence of
mixed with 1ml of conc.HNO3 precipitate/colouration aromatic compound
and 1ml of conc.H2SO4. heated No yellow colouration Presence of
on boiling water bath for aliphatic compound
15min., cooled and poured into
259ml of cold water
7. Test for saturation or unsaturation
A To about 0.1g of the substance, Decolouration Presence of
1ml bromine water is added and unsaturated
shaken well compound
No decolouration Presence of
unsaturated
compound
B To about 0.1g of the substance, Decolouration of pink Presence of
1ml Na2CO3solution and 1% colour unsaturated
KmnO4 solution dropwise compound
Presence of
No decolouration unsaturated
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compound
8 Action of sodium bicarbonate
To about 0.1g of the substance, Brisk effervesence Presence of
2ml of saturated NaHCO3 with evolution of CO2 carboxylic acid
solution is added which turns lime
water milky
9 Action of dil.HCl
To about 0.5g of the substance, Dissolution with Presence of basic
2ml of dil.Hcl is added, shaken precipitation on functional groups
well adding 10% NaOH such as amine
dropwise
10 Action of sodium hydroxide
To about 0.5g of the substance, Dissolution with Presence of
2ml of 10% NaOH, shaken well precipitate on addition aromatic
of conc.Hcl

Dissolves with strong

yellow colour which Presence of
disappears by addition aromatic nitro
of conc.HCl compound or
aldehyde
Evolution of NH3 gas

Solution turns yellow Presence of amide

or brown on boiling or ammonium salts
Presence of sugar or
Oily globules with aliphatic aldehyde
aniline smell on
boiling Presence of anilide

Oily layer that

disappears on boiling: Presence of ester or
white precipitate on amide of aromatic
acidification with acid
conc.HCL and
cooling

No charecteristic Presence of nitro

change in cold or compound ketone
boiling condition or amine

Test for phenols

A Phthalic anhydride test:
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To about 0.1g substance, 0.1g Pink colour Presence of

phthalic anhydride and 3 droops monohydric phenol
conc.H2SO4 are sdded. Heated Greenish yellow Presence of
gently for 2-3 mins and poured colour dihydric phenol
into 20ml dil.NaOH solution
B Dye test
About 1ml aniline is dissolved Orange dye Presence of phenol
in 4ml dil.Hcl, the solution
cooled under tap or in ice & Scarlet or red dye Presence of
added NaNO2 solution naphthol or
dropwisw. To this solution 0.5g resorcinol
of substance dissolved in 5ml
dil.NaOH solution is added
slowly
C Libermanns test
About 0.1g substance is mixed Red solution becomes Presence of
with 0.5g NaNO2 in a dry test deep blue with excess phenol(not
tube. Heated for 2mins & NaOH substituted phenols)
cooled. 5 drops of conc.H2SO4
is added. Bluish green solution
is poured into 30ml water in a
beaker and NaOH added in
exces
Test for Ester
A Hydrolysis Disappearance of Presence of ester
To sbout 0.2g of substance, pleasant odour
added 2ml 10% NaOH, boiled
for 3mins
B Precipitation of acid White precipitate Presence of
The reaction mixture from the aromatic ester
above test is cooled and 1ml
conc.HCL added

C Test with phenolphthalein

To about 0.2g substance, added Disappearance of pink Presence of ester
2 drops of phenolphthalein & 3 colour(on heating)
drops NaOH & heated
Test for aldehyde and ketone
A Borsches test
To about 0.1g substance, 1ml Yellow or orange Presence of
borsches reagent is added, precipitate aldehyde or ketone
boiled, conc.HCL added and
1ml of water
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B Phenyl hydrazine test

About 0.1g substance is mixed Yellow precipitate Presence of
with 1ml water or ethanol and aldehyde or ketone
1ml phenyl hydrazine. Heated Yellow crystals Presence of
on boiling water bath for carbohydrate
10mins
C Semicarbazide test Presence of
To about 0.2g substance added Colourless crystals aldehyde or ketone
a mixture of 0.2g semicarbazide
hydrochloride, 0.5g sodium
acetate & 2ml water. 7ml of
alcohol is added to get clear
solution, heated for 2mins &
cooled
D Schiffs reagent test Immediate or red Presence of
To about 0.1g substance added colour aldehyde
3-4 drops of schiffs reagent, Slow pink colouration
shaken well Presence of
aliphatic ketone
E Fehlings solution test Presence of
To about 0.1g substane added Red precipitate aldehyde or aldose
1ml each of fehlings solution
A&B and boiled
F Tollens reagent Black precipitate or Presence of
About 0.1g substane added 1ml bright silver mirror aldehyde or aldose
tollens reagent. Heated on
water bath for 5-10 mins

G Legal test Presence of ketone

To about 0.1g substane added Orange colour turns
2ml water. 5 drops of sodium purple
nitroprusside solution 5 drops
NaOH and 5 drops of glacial
aetic acid
Test for carbohydrates
Molischs test: To about 0.1g Deep violet ring at the Presence of
sudstance in 1ml water, added junction of two layers carbohydrate
1ml alcoholic -naphthol & spreading of colour
solution, shaken well & an standing
conc.H2SO4 added dropwise
through the sides
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Test for amines

A Dye test
About 0.1g substance is Scarlet red dye Presence of primary
dissolved in 2ml dil.HCL, amine
cooled under tap or ice & added
a mixture of 1ml of NaNO2
solution and 1ml of -naphthol
dissolved in NaOH
B Acetylation:
To about 0.1g substance added White crystals Presence of primary
1ml glacial acetic acid and amine
0.5ml acetic anhydride, heated
gently for 3-5mins & poured
into 20ml of water in a beaker
C Libermanns test:
About 0.1g substance is Red solution turns Presence of
dissolved in 3ml ethanol. To blue secondary amine
this 1ml of conc.HCL added,
cooled under tap or ice & 1ml
NaNO3 solution added. Shaken
well, either layer separated and
evaporated carefully. 5 drops
phenol added to the residue,
warmed, cooled and added 5
drops of conc H2SO4 & poured
into 20ml of dil NaOH solution
D Quartenary salt test
To about 0.1g substance added White crystalline solid Presence of tertiary
1ml CH3I solution, shaken amine
well, heated gently & allowed
to stand for 2-5 mins
test for amides
A Urea nitrate test
To about 0.1g substance added White precipitate Presence of diamide
1ml water amd 1ml
conc.HNO3, shaken well
B Urea oxalate test
About 0.1g substance is shaken White precipitate Presence of diamide
with 3ml of saturated oxalic
acis solution
C Biuret test
About 0.2g substance is Violet colour Presence of diamide
strongly heated in dry test tube
[Type text]

till NH3 smell ceases. Cooled

and added 1ml water, 4 drops
each of CuSO4 & NaOH
solution
Test for nitro group
Mulliken & Barkers test
About 0.1g substance is dissolved in 2ml of ethanol and sdded each of NH4CL & Zn
dust, heated for 3mins & allowed to stand for 3mins & filtered. Filte is divided into two
parts
A To one part 2ml tollens reagent Black precipitate or Presence of nitro
is added and heated ina water bright silver mirror compound
bath
B To other part, added each 1ml Red precipitate Presence of nitro
of fehlings solution A&B. compound
heated for 2mins
C Reduction
To about 0.3g substance added Positive observations Presence of nitro
5ml conc.HCL & a piece of Sn. for amines compound
Heated for 3-5mins and KOH
solution added till it becomes
alkaline. Separated loquid is
tested for amines
Test for anilides
A About 0.1g substance is
dissolved in 3ml glacial acetic Pale yellow Presence of anilide
acid and added 1ml of Br2 in precipitate
acetic acid. Shaken well &
poured into 10ml of water
B Dye test
About 0.1g substance is heated Scarlet red dye Presence of anilide
with 3ml conc.HCL for 2-3
mins, cooled under tap or in ice
heated with NaNO2 solution &
- naphthol solution in 2ml
NaOH
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METHODOLOGY OF FILTRATION AND RECRYSTALLIZATION

FILTRATIONS:

GRAVITY FILTERATION

Gravity filtration is the method of choice to remove solid impurities from an organic
liquid. The impurity can be a drying agent or an organic liquid. The impurity can be be a
drying agent or an undesied product or left over reactant. Gravity filteration can be usede
to collect solid product, although generally vaccum filteration is used for this purpose
because it is faster.

A filteration precedure called hot gravity filteration is used to separate insoluble

impurities from a hot solution. Hot filterations require filter paper and careful attention to
the prcedure to keep the appararus warm but covered co that solvent does not evaporate.

SUCTION FILTERATION:

Crystals may be collected from a solution by filteration on a funnel. A partial

vaccum is created in the filter flask bya water aspirator (water flowing past a small hole
in a pipe draws in gas from the hose to the filter flask. This reduces the pressure creating
the partial vaccum.) the filter paper should be placed in the funnel and wetted before
starting the filteration to help seal it to the funnel. Ensure that the filter flask is clamped
to the resort stand and the rubber stopper holding the funnel is tight in the filteration
flask. Turn on the water to the aspirator to create a reduced pressure in the filteration
flask. Swirl the last of the solution in the flask to help transfer any remaining crystals into
the funnel. Use the spatula to transfer crystals that are missed. Wash the crysta;s as
instructed in the lab manual andthen compact with a spatula to push out as much as
possible. Leave the crystals on the funnel with the aspirator running to further dry the
crystals. Check the lab manual for any drying instructions.

Recrystallization:

The organic compounds are purified by Recrystallization from a suitable solvent

which can dissolve a large quantity of the substance at high temperature and deposit the
same when cooled.

A nearly saturated solution of the impure substance is prepared in a hot solvent. It

is decolourised with animal charcoal and filtered while hot. The filterate on cooling
deposits crystals of the pure substance.
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