Qualitative analysis of carbohydrate

by Molisch Test.
 Molisch Test

Objectives of Molisch Test

•To detect the presence of carbohydrates in a given

sample.
•To distinguish carbohydrates from other biomolecules .

2
 Molisch Test
Principle of Molisch’s test:

Molisch’s test is a general test for all carbohydrates.

In this test, carbohydrates when reacted with conc.
H2SO4 get dehydrated to form furfural and its
derivatives.
When monosaccharide are treated with conc
H2SO4 or conc HCl, -OH group of sugar are
removed in the form of water and furfural is formed
from pentose sugar and hydroxymethyl furfural is
formed from hexose sugar. These products reacts
with sulphonated α- naphthol to give a purple
(violet red) colored complex.

3
 Molisch Test

Chemicals and Glassware’s:

1% alcoholic ∝-naphthol, conc. sulphuric acid, test sample,

test tubes, test tube stand, test tube holder,
Reagent preparations:
Molisch reagent was prepared by dissolving 3.75gm of ∝-
naphthol in 25ml of ethanol.

4
 Molisch Test

Procedure of Molisch Test

1.Take 2 ml of each distilled water and test sugar solutions in
four test tubes separately.
2.Add two drops of Molisch reagent to each tube.
3.Hold the test tube in an inclined position and gently add 1 ml
concentrated H2SO4 along the wall of the test tube. Do not mix
the acid with the solution. A black ring may form if
concentrated acid is not added slowly as the heat generated from
the reaction can char the carbohydrates.
4.Observe the test tube for the formation of a purple-colored
ring at the layer between the solution and the acid.
5
 Molisch Test

Result discussion:
At the whole we conclude that there is
the formation of purple color ring at the
junction between sulphuric acid and test
solution. The acid remains above the test
solution due to its high density. The
absence of color indicates a negative
result