Professional Documents
Culture Documents
Qualitative Tests For Proteins
Qualitative Tests For Proteins
ABSTRACT
This experiment used non-fat milk as the source of the protein isolate, casein. After
isolation and hydrolysis of the protein, qualitative tests were performed on both the protein
isolate and hydrolysate. The experiment aims to enable the students to perform qualitative
tests and explain the principles involved in each of the test. Out of the 10 qualitative tests,
the isolate tested positive for the following tests: Biuret, Ninhydrin, Xanthoproteic,
Sakaguchi, Fohls, Test for Amides, and Paulys. On the other hand, the hydrolysate tested
positive for the following tests: Ninhydrin, Xanthoproteic, Fohls, Test for Amides, and
Paulys. This indicates that acid hydrolysis may have destroyed or affected some of the
amino acid components of casein. Thus, qualitative tests alone are insufficient in identifying
the amino acid components of a protein. Factors such as cross-contamination of reagents
and the effect of acid hydrolysis are some sources of errors that may render the tests
inaccurate.
NaNO2 solution were mixed Test for (+) yellow (+) dirty
into each test tubes. 5 drops Amide soln yellow soln
of 10% Na2CO3 were then RB RB
added. litmus litmus
Paulys (+) orange (+) red soln
soln
RESULTS AND DISCUSSION
Table 1 below shows the results
Biuret Test is a tests for the
of the qualitative tests of intact casein
presence of peptide bonds. The
and hydrolyzed casein. These tests
principle involved is the formation of
help identify the amino acid
coordination complexes with copper
constituents of the protein through
(II). Because hydrolysis destroys the
color reactions with the different
primary structure of proteins, peptide
functional groups.
bonds are not present in the
hydrolysate.[6] Thus, the intact protein
Table 1. Qualitative Color Reactions of
gives a positive result while the
Intact Casein and Hydrolyzed Casein
hydrolysate gives a negative result.
Color Intact Acid
Reaction Protein Hydrolysis
Biuret (+) purple (-) black
soln soln
& ppt
Ninhydrin (+) violet (+) dark
ppt violet
soln & ppt Figure 1. Coordination complex
Xanthoprot (+) yellow (+) orange formation of copper (II)
eic ppt soln
& soln
Millons (-) white ppt (-) brown
soln
Hopkins- (-) white ppt (-) brown &
Cole colorless
layered soln
w/ brown
ring
Sakaguchi (+) light (-) brown
yellow soln ppt and
soln
Nitroprussid (-) yellow (-) yellow
e soln soln w/
brown ppt
Fohls (+) (+) brown
brownish ppt & soln Figure 2. Biuret Test on intact protein
black soln & (left) and hydrolysate (right)
ppt
PH-BIOCHEM Expt 2 2FPH Group 1
4