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Test Biuret Test

Reagents CuSO4 sol'n, diluted NaOH

Test for Intact proteins and protein hydrlysates (at least tripeptide, 2 peptide bonds

Result Pink to violet blue color

Reaction/Principle Complexation Form. Of coordination complex Cu+2 and 4 N atoms (2 from each 2 polypeptide chain)

Ninhyrin Test

1,2,3-indanetrione monohydrate or triketohydintene hydrate, ethanol Conc. HNO3, conc. NaOH

Alpha- amino group (usually a general test for amino acids)

Xanthoproteic Test

For W,F,Y (aromatic except for H)

Blue to blue-violet Oxidative decarboxylation color & deamination followed by (proline:hydroxypr condensation oline gives a yellow color) Yellow sol'n/ppt. Nitration via SEAr with conc. HNO3, orange with excess NaOH

Millon's rgt.(Hg2+ & Hg22+nitrates& nitriles/Hg(NO3)2 in conc. HNO3, w/ trace of HNO2/HgSO4,H2SO4 ,NaNO2) Hopkins-Cole Test Glyoxylic acid (Mg powder, oxalic acid, HOAc), conc. H2SO4 Millon's Test

Phenolic group of Y

Old rose/flesh to red ppt.

Complexation(mercuration & nitration or nitrosation/complexation of nitrohydroxyphenyl derivatives with Hg2+)

Indole group in W

Pink to violet interface

Reduction of oxalic acid to glyoxolic acid and acidcatalyzed condensation of two tryptphans w/glyoxolic acid

Sakaguchi Test

Alpha-naphthol, NaOBr, NaOH, urea(to stabilize color & destroy excess OBr ions)

Un-or monosubstituted guanidines in R

Red to red-orange Complexation (basecolor catalyzed condensation of alpaha-naphthol w/ the guanidine group of R)

Fohl's Test


Sulfur-containing C-C)


Brown to black ppt.

Degradation & substitution rxn to form PbS

Nitroprusside Test Test for Amide Pauly's Test

Na2Fe(CN)3 in dil. NH3 NaOH Diazo rgt. (1% sulfosalicylic acid with 5% NaNO2), 10% Na2CO3

For cys 10,20,30 amides and nitriles ; peptide bonds Detects tyr and his

Red coloration Red litmus to blue Red coloration

Complexation rxn Basic hydrolysis Formation of azo dyes