Qualitative Color Reaction Biuret Test

Test Presence of a Peptide Bond

Reagent CuSO4 + NaOH

Ninhydrin Test

a-amino acid

Ninhydrin

Xanthoproteic Test

Test for aromatic Amino acids

HNO3 , NaOH

Principle formation of coordination complex of Cu2+ and four nitrogen atoms oxidative decarboxylation & deamination followed by condensation nitration of aromatic rings via SEAr

Positive Result Purple solution

-Blue-Violet Color -Yellow for Proline - yellow color of solution with HNO3 - orange color of solution with NaOH Red Color

Millons Test

Test for Tyrosine

salt of Hg dissolved in HNO3

Hopkins-Cole Test

Test for Tryptophan

glacial CH3COOH, glyoxylic acid, concd H2SO4 -naphthol, NaOBr, NaOH, urea

Sakaguchi Test

Test for Arginine

Nitroprusside Test Fohls Test Test for Amide Pauly Test

Test for S-containing amino acids Test for S-containing amino acids Asparagine and Glutamine Test for Histidine and Tyrosine

NaOH, Nitroprusside NaOH, (CH3COO)2Pb NaOH sulfanilic acid in NaNO2 solution,Na2CO3

mercuration & complexation of nitrohydroxyphenyl derivatives with Hg2+ reduction of oxalic acid to glyoxylic acid and acid-catalyzed condensation of two tryptophans with glyoxylic acid base-catalyzed condensation of -naphthol with the guanido group of Arg Complexation Reaction degradation & substitution reaction to form PbS Basic Hydrolysis Formation of azo dyes

Purple Interface

Red Color

Red Color dark brown / black precipitate Evolution of gas (presence tested in a litmus paper) Red color

Color Reaction Biuret Ninhydrin Xanthoproteic Millons Hopkins-Cole Sakaguchi Nitroprusside Fohls Test for amide Pauly

Intact Protein + +

+ + + + + + +

acidic + +/- Tyrosine + Phenylalanine - Tryptophan +/+ + Methionine +/- Cysteine + Methionine +/- Cysteine + Histidine +/- Tyrosine

Protein Hydrolysate basic + +

enzymatic + +/+

+ + + + +

+ + + + + + +