Test Reagents Test for Result Reaction/Principle

Biuret Test CuSO4 sol'n, diluted Intact proteins and Pink to violet blue Complexation
NaOH protein hydrlysates (at color Form. Of coordination
least tripeptide, 2 complex Cu+2 and 4 N
peptide bonds atoms (2 from each 2
polypeptide chain)

Ninhyrin Test 1,2,3-indanetrione Alpha- amino group Blue to blue-violet Oxidative decarboxylation
monohydrate or (usually a general test color & deamination followed by
triketohydintene for amino acids) (proline:hydroxypr condensation
hydrate, ethanol oline gives a yellow
color)
Xanthoproteic Conc. HNO3, conc. For W,F,Y (aromatic Yellow sol'n/ppt. Nitration via SEAr
Test NaOH except for H) with conc. HNO3,
orange with excess
NaOH

Millon's Test Millon's rgt.(Hg2+ & Phenolic group of Y
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
Old rose/flesh to Complexation(mercuration
red ppt. & nitration or
nitrosation/complexation of
nitrohydroxyphenyl
derivatives with Hg2+)
Indole group in W Pink to violet Reduction of oxalic acid to
interface glyoxolic acid and acid-
catalyzed condensation of
two tryptphans w/glyoxolic
acid

Sakaguchi Test Alpha-naphthol, Un-or monosubstituted Red to red-orange Complexation (base-

NaOBr, NaOH, guanidines in R color catalyzed condensation of
urea(to stabilize alpaha-naphthol w/ the
color & destroy guanidine group of R)
excess OBr ions)

Fohl's Test Pb(CH3COO)2 , Sulfur-containing (C, Brown to black ppt. Degradation & substitution
NaOH C-C) rxn to form PbS

Nitroprusside Test Na2Fe(CN)3 in dil. For cys Red coloration Complexation rxn
NH3
Test for Amide NaOH 10,20,30 amides and Red litmus to blue Basic hydrolysis
nitriles ; peptide bonds
Pauly's Test Diazo rgt. (1% Detects tyr and his Red coloration Formation of azo dyes
sulfosalicylic acid
with 5% NaNO2),
10% Na2CO3