Experiment 1: Detection of carbohydrate in an unknown solution using Molisch’s Test

Objective: To identify carbohydrate from other biomolecules

Principle: Molisch’s test is a general test for all carbohydrates. In this test, carbohydrates when reacted
with concentrated H2SO4 get dehydrated to form furfural and its derivatives. When monosaccharide are
treated with concentrated H2SO4 or concentrated 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.
Requirements:
1. Test solution: 5 % D-Glucose
2. Molisch’s reagent (5 % α naphthol in ethanol)
3. Concentrated H2SO4
4. Dry test tubes & stand
5. Pipettes
Procedure:
1. Take 2ml of sugar sample in dry test tube.
2. Add 2-3 drops of Molisch’s reagent to the solution.
3. Gently pipette 1ml concentrated H2SO4 along the wall of the tube to form distinct layers.
4. Observe color change at the junction of two layers.
5. Appearance of purple color indicates the presence of carbohydrates.
Chemical Reaction(s):

Result:
Positive Molisch’s Test will produce purple (or violet red) color complex
Fig 1: Negative & Positive Molisch’s Test