BIOCHEMISTRY LABORATORY | Pam Garcia

COLOR REACTIONS

TEST PURPOSE REAGENT POSITIVE NEGATIVE PRINCIPLE MECHANISM

S RESULT RESULT

Biuret Test Analysis of 2.5M NaOH Pink to violet Blue Complexation Biuret reacts with
proteins solution solution copper ions, forming a
with at 0.01M (color of violet complex (Biuret
least two CuSO4 Pink: the complex)
peptide PEPTONES reagent)
bonds in its
chain Purple:
Peptide
bonds
Sakaguchi Detects the 10% NaOH Red to red- Yellow R residue reacts Base-catalyzed
Test presence orange solution with alpha- condensation of alpha-
of 0.02% solution naphthol and naphthol w/ the
guanidine naphthol NaClO to give guanidine group in R;
group (in solution chromogen; Hypochlorite oxidizes
R) NaOH provides an the guanidine group and
NaClO alkaline pH to the alpha-naphthol
solution. At this condenses the product
pH, guanidine producing a red solution
group combines
with naphthol

Ninhydrin General 0.1% Blue to blue- Alternativ Alpha amino Oxidative deamination
Test test for Ninhydrin violet e result: groups react with and decarboxyl-ation;
compounds solution solution yellow ninhydrin to form reduction of ninhydrin
with free (Ninhydrin solution aldehyde,
alpha- in 95% (imino hydrindantin,
amino ethanol) acids) ammonia and
groups carbon doxide.
One molecule of
BIOCHEMISTRY LABORATORY | Pam Garcia

hydrindantin,
ninhydrin and
ammonia
complex forms
Ruhemanns
purple
Xanthoprot Detects the Conc. HNO3 White White precipitate Nitration of aromatic
eic Test presence Conc. precipitate forms upon rings via electrophilic
of aromatic NaOH in yellow adding nitric acid, aromatic substitution
ring solution the benzene ring
structures after of AAs undergo
addition of nitration with hot,
conc. HNO3; strong HNO3,
yellow producing a
solution yellow color in an
deepens to acidic medium
an orange then deepens to
color after an orange color
addition of upon treatment
NaOH with NaOH (to
neutralize)
Hopkins- Test for the Hopkins Presence of The protein Reduction of oxalic acid
Cole Test presence reagent pink to violet solution is mixed to glyoxylic acid & acid-
(Glyoxylic of W (powdered interface No with Hopkins-Cole catalyzed condensation
reaction) containing magnesium interface reagent, upon of Tryptophan with
an indole , oxalic (Interface: addition of glyoxylic acid
group acid, & the junction sulfuric acid, a
acetic acid) between the ring appears at
two layers) the junction of
Conc. two liquids
H2SO4
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TEST Intact Acid Base

casein hydrolyzat hydrolyzate
e
Biuret Test + - -

Sakaguchi + + -
Test
Ninhydrin - + +
Test
Xanthoprot + + +
eic Test
Hopkins- + - +
Cole Test