BIOCHEMISTRY

AIM: To perform the qualitative analysis and identify the given sample of
carbohydrate (fructose).

REQUIREMENTS: Test tube, holder, measuring cylinder, spatula.

THEORY:
EXPERIMENT

Molischs Test:
To 2ml of the sugar
solution add 2
drops of molischs
reagent (5% of xnaphthol in
absolute alcohol)
and mix well. Then
add 2 ml of conc.
H2SO4 at the sides
of the test tube
slightly in an
inclined plane.
Then erect the
testube slowly and
note the junction
of two layers.
Iodine Test: To
2ml of the sugar
solution add 5
drops of GAA and
mix. Then add 3
drops of N/10
iodine drop by
drop, mixing after
addition.
Benedicts test:
To 2ml of
Benedicts
reagents, add 1ml
of sugar solution
and keep it in the
water for 2
minutes.
Fehlings test:
Add 1ml of

OBSERVATION
A violet or purple
covering is seen at the
junction of the two
liquids.

INFERENCE
It is a general test for
carbohydrates whether
free or bound form. With
H2SO4 furfural or its
derivatives are readily
formed by the removal
of water molecule.
Furfural formed
condenses with xnaphthol and gives a
coloured compound.
Glucose / fructose/
maltose/lactose/starch
being a carbohydrate
answer this test.

No colour change was

observed.

Formation of yellow ppt.

Appearance of red ppt.

Development of yellow
ppt.

This is a specific test for

polysaccharide.
Monosaccharide and
disaccharide donot give
this test. Polysaccharide
is absent.

It is a sensitive reduction
test indicating the
presence of reducing
sugar. Reducing
monosacchride may be
present.
Presence of reducing
sugar confirmed.

Fehlings Reagent
to 1 ml of test
solution. Mix
thoroughly and
keep the tt in
boiling water bath.
Picric Acid Test:
To 5ml of the test
solution add 2-3 ml
of saturated picric
acid solution and
about 1ml of 10%
of Na2CO3. Warm.

Presence of reducing
sugar confirmed.

EXPERIMENT

Tommers Test:
To 3ml of test
solution add 2ml of
tommers reagent.
Boil for 2mins and
cool.
Barfoeds Test:
To 2 ml of
barfoeds reagent
(cupric acetate in
acetic acid) add 1
ml of sugar
solution and keep
in boiling water
bath for 2 minutes.

OBSERVATION

INFERENCE

Yellow red ppt was

formed.

Presence of reducing
sugar is confirmed.

Appearance of red ppt.

Sample may be
monosacchride.

Cherry red colour was

observed.

Ketohexoses is present.

Yellow ppt was formed.

This is the test to identify

the reducing sugar and to
confirm the type of sugar.

Needle like crystals.

Fructose is present.

Seliwanoffs
Test: Add 1 ml of

the sugar solution

to 2 ml of
seliwanoffs
reagent(0.1%
resorcinol in 50%
HCL) and warm in
boiling water bath
for few minutes.
Osazone Test:
To 3ml of sugar
solution add a
pinch of phenyl
hydrazine
hydrochloride,
sodium acetate
mixture. Mix well
and heat in the
boiling water bath
for 30 minutes.
Cool the content
and view the
crystals under
microscope.

RESULT: The given carbohydrate is a monosaccharide reducing ketosugar

FRUCTOSE.