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EGG ALBUMIN SOLUTION No color changes occurred.

Remained colorless.
Acted as constant.

ASPARTAME No color changes occurred.

Remained colorless.
Acted as constant.

EVAPORATED MILK No color changes occurred.

Remained white in color.
Acted as constant.

Egg & Milk: Separated and Egg & Milk: Positive

A. HOPKINS – COLE TEST formed two layers and a ring was
observed at the center. Colors
were purple and whitish cloud. Aspartame: Negative
Aspartame: None.
Egg & Milk: The color changed Egg & Milk: Positive
into violet.
Aspartame: Colorless, Aspartame: Negative
Egg & Milk: The color changed Egg & Milk: Positive
C. NINHYDRIN TEST into purple.

Aspartame: Crystal clear blue. Aspartame: Negative

Egg: Red color was observed, the Egg: Positive
D. SAKAGUCHI TEST color went to the bottom.
Milk: Red color stayed at the Milk: Positive
Aspartame: Light green was Aspartame: Negative
Orange colored lower layer,
E. XANTHOPROTEIC TEST yellowish cloud top layer.
Aspartame: Clear with a less
pigmented yellow liquid.

During the Hopkins- Cole Test, egg and milk did changed in color after the concentrated sulfuric
was added, a purple ring appeared between the two layers. This test is used for detecting the presence
tryptophan in proteins. Among protein amino acids, only tryptophan reacted with glyoxylic acid in the
presence of conc. H2SO4 and gave a purple complex. On the other hand, the Aspartame has no reaction,
remained transparent.

In Biuret test, egg and milk turned into violet. The reagents used were CuSO4 and NaOH. Sodium
Hydroxide used to raise the pH of the solution to alkaline level. The complex of Cu2 ions and nitrogen
atoms made the color CuSO4 solution changed into violet. Color changed is dependent on the number of
peptide bonds in the solution, the more protein, the more pigmented the color change. The test showed
the presence of proteins in the following food. Moreover, the aspartame remained colorless.

The Ninhydrin test results were accurate. The milk solution and egg albumin changed color into
purple. These were because in pH range 4 to 8, all amino acid reacted with ninhydrin, a powerful
oxidizing agent gave a purple outcome. Hence, proteins were present. Aspartame gave a crystal, clear
blue outcome.

During the Sakaguchi test, a red-colored complex formed in both the egg and the milk. The
aspartame produced a shade of light green. Sakaguchi test is specific for Arginine. The test is positive for
containing the guanidine group in Arginine. Guanidine group present in the amino acid reacted with α-
Napthol and alkaline hypobromite which gave a red-colored complex. Hence, proteins containing
Arginine were present.

In Xanthoproteic test, orange-colored and yellowish cloud color was observed in both the egg
and milk. The changed in color was due to the high temperature of which the Phenylalanine, tyrosine
and tryptophan reacted with the conc. HNO3, formed nitro-compounds which were yellow in color, it
turned orange-colored in an alkaline medium. Furthermore, amino acids containing the benzene ring
(aromatic amino acids) were present.

Faculty of Medicine, Biochemistry Department. (n.d). Biochemistry Practical: Proteins & Amino Acids
Identification. Retrieved from

Izneera P. (n.d). DOC Lab Report. Experiment B: Qualitative Test For Proteins. Retrieved from