Biomolecule Tests

Molisch’s Test
Molisch’s test is a chemical test which is used to check for the presence of carbohydrates in a given analyte.
Molisch’s test involves the addition of Molisch’s reagent (a solution of ””-naphthol in ethanol) to the analyte and the
subsequent addition of a few drops of concentrated H2SO4 (sulphuric acid) to the mixture.
The formation of a purple or a purplish-red ring at the point of contact between the H2SO4 and the analyte +
Molisch’s reagent mixture confirms the presence of carbohydrates in the analyte.

Ninhydrin Test
The ninhydrin test is a chemical test which is used to check whether a given analyte contains amines or á-amino
acids. In this test, ninhydrin (a chemical compound with the formula C9H6O4; IUPAC name: 2,2-dihydroxyindane-1,3-
dione) is added to a test solution of the analyte. The development of a deep blue colour indicates the presence
of ammonia, primary/secondary amines, or amino acids in the analyte.

Xanthoproteic Test
Xanthoproteic test is specific for protein containing aromatic amino acids. The benzene ring in the amino acids is nitrated
by heating with nitric acid and forms yellow nitro-compounds which turns to orange colour with alkali.

Millon’s Test
Millon’s reagent is an analytical reagent used to detect the presence of soluble proteins. A few drops of the reagent
are added to the test solution, which is then heated gently. A reddish-brown coloration or precipitate indicates the
presence of tyrosine residue which occur in nearly all proteins.
Millon’s test is not specific for proteins; it also gives a positive test for other compounds containing the
phenol functional group. Therefore, the biuret test or the ninhydrin reaction are used along with it to confirm the
presence of proteins.

Seliwanoff’s Test
Seliwanoff’s test is a chemical test which distinguishes between aldose and ketose sugars. This test relies on the
principle that, when heated, ketoses are more rapidly dehydrated than aldoses. When added to a solution containing
ketoses, a red color is formed rapidly indicating a positive test. When added to a solution containing aldoses, a slower
forming light pink is observed instead.

Barfoed Test
Barfoed’s test is a chemical test used for detecting the presence of monosaccharides. It is based on the reduction
of copper(II) acetate to copper(I) oxide (Cu2O), which forms a brick-red precipitate.
The test is similar to the reaction of Fehling’s solution to aldehydes.

Biuret Test
The biuret test, also known as Piotrowski's test, is a chemical test used for detecting the presence of peptide bonds. In
the presence of peptides, a copper(II) ion forms mauve-colored coordination complexes in an alkaline solution.