Molisch's test

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Molisch's test (named after Austrian botanist Hans Molisch) is a sensitive chemical test for the presence of carbohydrates, based on the dehydration of the carbohydrate by sulfuric acid to produce an aldehyde, which condenses with two molecules of phenol (usually α-naphthol, though other phenols (e.g. resorcinol, thymol) also give colored products), resulting in a red- or purple-colored compound.

Procedure

The test solution is combined with a small amount of Molisch's reagent (α-naphthol dissolved in ethanol) in a test tube. After mixing, a small amount of concentrated sulfuric acid is slowly added down the sides of the sloping test-tube, without mixing, to form a bottom layer. A positive reaction is indicated by appearance of a purple ring at the interface between the acid and test layers.

Reaction

All carbohydrates – monosaccharides, disaccharides, and polysaccharides – should give a positive reaction, and nucleic acids and glycoproteins also give a positive reaction, as all these compounds are eventually hydrolyzed to monosaccharides by strong mineral acids. Pentoses are then dehydrated to furfural, while hexoses are dehydrated to 5-hydroxymethylfurfural. Either of these aldehydes, if present, will condense with two molecules of naphthol to form a purple-colored product, as illustrated below by the example of glucose:

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