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EXPERIMENT No.08

Aim: To test for the presence of Carbohydrates in the given organic compound.

Material Required: Glucose, Sucrose, Molisch’s Reagesnt, Fehling’s reagent, Benedict’s reagent, Tollen’s
Reagent, test tube.

S.No. Experiment Observation Inference

MOLISCH’S TEST:

1-2ml of aqueous solution of carbohydrate, few A red violet ring is

drops of molisch’s reagent (1% alcoholic Presence of
1 modered at the junction
solution of 1- naphthol) was added. Then add 1 Carbohydrate
of two layer
ml of conc. H2SO4 slowly along the side of test
tube.
BENEDICT’S TEST:

To 1-2 ml of aqueous solution of carbohydrate in

Presence of
2 a test tube, 1-2 ml of Benedict’s reagent was A reddish ppt. indicates
Carbohydrate
added. Then the test tube was kept in a boiling
water bath for 10 mins.

FEHLING TEST:

2ml of aqueous solution of the carbohydrate is Presence of

3 taken and to this 1-2ml each of Fehling solution A reddish ppt. indicates
Carbohydrate
“A” and “B” is added. Now the test tube is kept
in a water bath for boiling.
TOLLEN’S TEST:

We take 2-3ml of aqueous solution of the A shining silver mirror

Presence of
4 carbohydrate in a test tube. Add to it 2-3ml indicates on the wall of
Carbohydrate
Tollen’s reagent. Keep this test tube in a water inner side of test tube
bath for 10 mins.

Chemical equation for Fehling Test:

COO-Na
|
(CHOH)4 + 3H2O+Cu2O↓
|
CH2OH
CHO
|
(CHOH)4 + 2Cu(OH)2 + NaOH Tartrate Ions

|
CH2OH

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Glucose Sod. Salt of gluconic acid

Chemical equation for Tollen’s Test:

AgNO3 + NH4OH NH4NO3 + AgOH

2AgOH Ag2O + H2O

Ag2O + 2NH4OH 2[Ag(NH3)2OH + 3H2O

(Soluble)

CHO COOH
| |
NH4OH
(CHOH)4 + Ag2O + NaOH
(CHOH)4 + 2Ag
| | (Silver mirror)
CHO COOH
Glucose Gluconic acid

Result: The given sample contain is Carbohydrate.

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EXPERIMENT No.09

Aim: To test for the presence of Proteins in the given organic compound.

Material Required: Egg albumin, 1% CuSO4 solution, Conc. HNO3, NaOH Solution, Ninhydrine solution,
Millon’s reagent and test tube.

S.No. Experiment Observation Inference

BIURET TEST:

2 ml of NaOH solution was added to the

Bluish violet colouration
1 dispersion of the substance in test tube, add ed 4- Presence of Protein
indicates
5 drops of 1% CuSO4 solution and the mixture
was warmed for about 5 min.

XANTHOPROTIC TEST:

2ml of egg albumin was taken in test tube and A yellow colouration
2 Presence of Protein
add 3-4 drops of conc. HNO3 was added to it indicates
then heated.

NINHYDRIN TEST:
Intense blue colouration
3 2ml of egg albumin was taken in test tube and Presence of Protein
indicates
add a few drops of ninhydrin Solution was
added to it. Then the contents were boiled.

MILLON’S TEST:
White ppt. changes to
4 Presence of Protein
2 drops of Millon’s reagent was added to 1-2 ml brick red on boiling
of egg albumin solution.

Result: The given sample contain is Proteins.

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EXPERIMENT No.10

Aim: To separate the coloured compounds present in the mixture of red and blue inks by ascending paper
chromatography and find their Rf values.

Apparatus: Gas jar, glass rod, filter paper strip (Whatman No. 1 filter paper), jas cover and a fine capillary
tube.

Material Required: A mixture of red and blue inks, alcohol and distilled water.

Procedure:
1. Take a Whatman filter paper strip (20 x 2cm) and draw a line with pencil above 3 cm from
one end. Draw another line lengthwise from the centre of the paper.

Spotting of the mixture

2. With the help of fine capillary tube, put a drop of the mixture of red and blue inks at the
point P. Let it dry in air. Put another drop on the same spot and dry again. Repeat 2-3 times,
so that the spot is rich in the mixture.
3. Suspend the filter paper vertically in a gas jar containing the solvent (eluent) with the help
of a glass rod in such a way that the pencil line ( and the spot) remains about 2cm above the
solvent level (50% alcohol + distilled water).
4. Cover the jar and keep it undisturbed. Notice the rising solvent along with the mixture of
red and blue inks. After the solvent has risen about 15cm you will notice two different
spots of blue and red colours on the filter paper.
5. Take the filter paper out of the jar and mark the distance that the solvent has risen on the
paper with a pencil. This is called the solvent front.
6. Dry the paper. Out pencil marks in the centre of the blue and red spots.
7. Measure the distance of the two spots from the original line and the distance of the solvent
from the original line.
8. Calculate the Rf values of the blue and red inks by using the formula:

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Rf = Distance travelled by the blue or red ink from the point of application
Distance travelled by the solvent from the original line

Observation and Calculations:

Distance travelled by
Distance travelled by
Substance the different Rf Values
solvent
components
3.5 cm (Red Ink) 5.2 cm 3.5 / 5.2 = 0.67
Red Ink + Blue Ink
5.0 cm (Blue Ink) 5.2 cm 5.0 / 5.2 = 0.96

Result:
1. Rf Value of Red Ink = 0.67
2. Rf Value of Blue Ink = 0.96

Precautions:
1. Use good quality pencil for drawing the reference line so that the mark does not dissolve in the solvent
in which the chromatography is carried out.
2. Always make use of a fine capillary tube.

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3. Keep the jar undisturbed and covered during the experiment.
4. A spot should be small and rich in mixture.
5. Allow the spot to dry before putting the strip in the jar.
6. Keep the strip erect. Do not let it to be curled.
7. Do not allow the spot to dip in the solvent.

EXPERIMENT No.11

Aim: To test for presence of Alcoholic group.

Test for the Alcoholic [R-OH] group

S.No. Experiment Observation Inference

SODIUM METAL TEST:

We added 1ml of a given liquid in a dry test

A brisk effervescence
tube. Then we added 1gm of anhydrous CaSO4 Presence of alcoholic
1 due to the evolution of
and shaked it to remove water. Now we filtered group
H2 gas
out the liquid to another clean dry test tube and
then added a small piece of sodium metal.

CERIC AMMONIUM NITRATE TEST:

Appearance of red Presence of alcoholic
2 We added 1ml of the given compound in a clean
colour group
dry test tube and then added a few drops of ceric
ammonium nitrate reagent and shake it.

Sodium Metal Test:

2R-OH + 2Na 2R-O- Na+ + H2↑

Alcohol Sodium Alkoxide

2CH3OH + 2Na 2CH3-O- Na+ + H2↑

Methanol Sodium Methoxide

Ceric Ammonium Nitrate Test:

2R-OH + (NH4)2Ce(NO3)6 [(ROH)2Ce(NO3)4] + 2NH4NO3

Alcohol ceric ammonium nitrate Red complex

2CH3-OH + (NH4)2Ce(NO3)6 [(CH3OH)2Ce(NO3)4] + 2NH4NO3

Red complex

Result: The given organic compound contains alcohol group.

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EXPERIMENT No.12

Aim: To test for presence of Phenolic group.

Test for the Phenol [Ar-OH] group

S.No. Experiment Observation Inference

LITMUS TEST:
Presence of phenolic
1 A drop of the given liquid was placed on the Colour changes to red
group
moist blue litmus paper

FERRIC CHLORIDE TEST:

Presence of phenolic
2 Violet colouration
1ml of neutral ferric chloride solution was added group
in a test tube with 2-3 drops of the liquid.

LIEBERMANN’S TEST:
First we observed a deep
We placed 2-3 crystals of sodium nitrite in test blue colour and after
tube and added 1g of given liquid then heated for adding water red colour Presence of phenolic
3
half a minute and allowed it to cool. Then we appears, after adding group
added 1ml of conc. H2SO4 and shaked it to mix NaOH solution blue
the contents. Then we added water. After that colour appeared.
added excess of NaOH Solution

Ferric Chloride Test:

FeCl3 + 6C6H5OH [Fe(OC6H5)6]3- + 3 HCl

Ferric Phenol Violet Complex
Chloride
Liebermann’s Test:

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Result: The given organic compound contains phenolic group.

EXPERIMENT No.13

Aim: To test for presence of Aldehydes group.

Test for the Aldehyde [-CHO]

S.No. Experiment Observation Inference

SCHIFF’S TEST:

0.5ml of the given compound in alcohol was Appearance of red Presence of aldehyde
1
taken in clean dry test tube. Then we added 1ml colour group
of schiff’s reagent.

TOLLEN’S TEST:

1ml of silver nitrate solution was taken in test

tube and 2-3ml of dil. NaOH was added. Brown A shining mirror on the Presence of aldehyde
2
ppt. formed. dil NH3 solution was added drop walls of the test tube group
wise until brown ppt. of silver oxide just
dissolved. Then given liquid was added and test
tube was warmed in water bath for 5 min.

FEHLING’S TEST:
Appearance of a red Presence of aldehyde
3 In a test tube with 1ml of each Fehling’s “A”
Precipitate group
and “B”. We added 4-5 drops of the given
compound and warmed it in water bath for 5
mins.

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Tollen’s Test:

2Ag(NH3)2+ + RCHO + 3OH- RCOO- + 2Ag↓ + 4NH3 + 2H2O

2Ag(NH3)2+ + CH3CHO + 3OH- CH3COO- + 2Ag↓ + 4NH3 + 2H2O
Acetaldehyde Acetate ion Silver

Fehling’s Test:

RCHO + 2Cu2+ + 5OH- Cu2O(s) ↓ + RCOO- + 3H2O

Aldehyde Fehling’s (Red)
Solution

Result: The given organic compound contains aldehyde group.

EXPERIMENT No.14

Aim: To test for presence of Ketone group.

Test for the Ketones [-CO-]

S.No. Experiment Observation Inference

m-DINITROBENZENE TEST:

In a clean dry test tube we added 0.5ml of the Appearance of violet

Presence of ketonic
1 given compound and added 0.1gm of m- colour which slowly
group
dinitrobenzene powder. Then we added 1ml of fades
dilute NaOH solution and then shaked it.

SODIUM NITROPRUSSIDE TEST:

We dissolved a crystal of sodium nitroprusside

Presence of ketonic
2 in about 1ml of distilled water in a clean test A Red Colouration
group
tube and then added 0.5ml of the given
compound after shaking we added NaOH
solution.

Sodium Nitroprusside Test:

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CH3COCH3 + OH- CH3COCH2- + H2O
2- -
[Fe(CN)5NO] + CH3COCH2 [Fe(CN)5NO.CH3COCH2]3-
Nitroprusside Ion Red colouration

Result: The given organic compound contains ketonic group.

EXPERIMENT No.15

Aim: To test for presence of Carboxylic group.

Test for the Carboxylic [-COOH]

S.No. Experiment Observation Inference

LITMUS TEST:
Presence of carboxylic
1 Colour changes to red
A drop of given liquid is put on a moist blue group
litmus paper.

SODIUM HYDROGEN CARBONATE

TEST:
Presence of carboxylic
2 A brisk effervescence
group
A pinch of NaHCO3 was added to 1ml of organic
liquid in a test tube.

ESTER TEST:

1 ml of ethyl alcohol and 1-2 drops of conc. Presence of carboxylic

3 A fruity smell
H2SO4 were added to the organic compound and group
heated in a water bath for 5 min. reaction
mixture was poured in a beaker containing water

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Litmus Test:

Sodium Hydrogen carbonate Test:

RCOOH + NaHCO3 RCOONa + CO2↑ + H2O

Carboxylic (effervescence)
Acid

Ester Test:
H SO
RCOOH + C2H5OH 2 4 RCOOC2H5 + H2O
Carboxylic (Ester)
Acid

Result: The given organic compound contains Carboxylic group.

EXPERIMENT No.16

Aim: To test for presence of primary amine group.

Test for the Amine [-NH2]

S.No. Experiment Observation Inference

AZO DYE TEST:

In 3 test tubes A, B and C, 0.2 gm of the

Compound in 2ml of dilute HCl and was
Presence of Amine
dissolved in test tube A. In test tube B we Formation of Red /
1 group (Primary
prepared an aqueous solution of sodium nitrite. Orange dye
aromatic)
In test tube C we dissolved 0.2gm of B-napthaol
in Dilute NaOH. After that we added sodium
nitrite solution to test tube A and resulting
solution to the test tube C.

CARBYLAMINE TEST:

In a test tube we added 0.2gm of KOH with 2ml

Presence of Primary
2 of ethanol. Than we warmed the test tube until Offensive smell
amine
the pallets dissolved. Then we added few drops
of chloroform and small amount of the given
compound and warmed it.

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Azo Dye Test:

C6H5NH2 + HNO2 + HCl C6H5N+ ≡ NCl- + 2H2O

Aniline Nitrous Acid (Benzene diazonium
chloride)

Carbylamine Test:

RNH2 + CHCl3 + 3KOH RNC + 3KCl + 3H2O

(Carbylamine
Isocyanide offensive smell)
Result: The given organic compound contains is amino group.

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