COLOR REACTION TEST

BIURET TEST detects the presence of peptide bonds. In the presence of peptides, a copper(II) ion forms
violet-colored coordination complexes in an alkaline solution. The Biuret reagent is made of potassium
hydroxide (KOH) and hydrated copper(II) sulfate, together with potassium sodium tartrate. The casein is
positive in Biuret test.

MOLISCHS TEST is a test for carbohydrates in general. This test is useful for identifying any compound
which can be dehydrated to furfural or hydroxymethyl furfural in the presence of H2SO4. The result
from the Molischs test is the formation of Violet ring and the casein is positive with this test.

MILLONS TEST detects the presence of Tyrosine. It is not specific for proteins (it actually detects
phenolic compounds), and so must be confirmed by other tests for proteins such as the biuret test and
the ninhydrin reaction. Consequently, any protein containing tyrosine will give a positive test of a pink to
dark-red colour. The result that we obtain in this solution is negative.
The Millon reagent is a solution of mercuric and mercurous ions in nitric and nitrous acids. The
red colour is probably due to a mercury salt of nitrated tyrosine

NINHYDRIN TEST is a test for Amine group, mainly primary Amine groups in Proteins, Peptones and
Amino acids. A protein has an amino terminal and a -COOH terminal. The ninhydrin will react with the
amino terminal giving a very light blue or violet color, more often than not you will see no color change.
Upon further heating there will be an increase in the intensity of the blue/violet. This is due to the heat
denaturing the protein, thereby exposing more -NH2 groups for the ninhydrin reagent to react with. Our
actual result from this test is positive having a very light blue color in the solution.
A ninhydrin solution is commonly used by forensic investigators in the analysis of latent
fingerprints on porous surfaces such as paper.
Amino acid containing fingermarks, formed by minute sweat secretions which gather on the
fingers unique ridges, are treated with the ninhydrin solution which turns the amino acid finger ridge
patterns purple and therefore visible.

REDUCED SULFUR TEST detects the presence of sulfur-containing proteins, specifically amino acids such
as methionine and cysteine. The positive result from this test should be a black precipitate. Since we
didnt finish heating our solution we were not able to get a positive result.

DENATURATION refers to any disruption in the secondary, tertiary or quaternary levels of protein
structure. Its positive result is coagulation or precipitation. Loss of biological activity is the most
significant consequence of denaturation.
BY HEAT: Heat can be used to disrupt hydrogen bonds and non-polar hydrophobic interactions. This
occurs because heat increases the kinetic energy and causes the molecules to vibrate so rapidly and
violently that the bonds are disrupted. Our result from this test is negative, the albumin become watery
probably because there wasnt enough heat.
Medical supplies and instruments are sterilized by heating to denature proteins in bacteria and
thus destroy the bacteria.

BY ALCOHOL: Alcohol coagulates all types of proteins except Prolamines. Alcohol denatures protein by
forming hydrogen bonds that compete with naturally occurring hydrogen binds in proteins. This process
is not reversible.
70% alcohol solution is used as a disinfectant on the skin. This concentration of alcohol is able to
penetrate the bacterial cell wall and denature the proteins and enzymes inside of the cell.
95% alcohol solution merely coagulates the protein on the outside of the cell wall and prevents
any alcohol from entering the cell. Alcohol denatures proteins by disrupting the side chain
intramolecular hydrogen bonding. New hydrogen bonds are formed instead between the new alcohol
molecule and the protein side chains.
BY INORGANIC ACIDS: Changes in pH can disrupt hydrogen bonds and salt bridges, causing irreversible
denaturation.
Hellers ring test is used to detect albumin in urine. Nitric acid. White precipitate.
BY HEAVY METALS: Heavy metal salts act to denature proteins in much the same manner as acids and
bases. Heavy metal salts usually contain Hg+2, Pb+2, Ag+1 Tl+1, Cd+2 and other metals with high atomic
weights. Since salts are ionic they disrupt salt bridges in proteins. The reaction of a heavy metal salt with
a protein usually leads to an insoluble metal protein salt.
This reaction is used for its disinfectant properties in external applications. For example AgNO3
is used to prevent gonorrhea infections in the eyes of new born infants. Silver nitrate is also used in the
treatment of nose and throat infections, as well as to cauterize wounds.
Mercury salts administered as Mercurochrome or Merthiolate have similar properties in
preventing infections in wounds.
This same reaction is used in reverse in cases of acute heavy metal poisoning.
BY ALKALOID REAGENTS: Alkaloid reagents such as Picric acid and Tannic acid from insoluble compounds
with protein. Alkaloidal reagents denature protein irreversibly by disrupting salt bridges and hydrogen
bond.
Tannic acid has been used extensively in the treatment of burns. It causes protein to precipitate
as a tough covering, reducing the water loss from the area and exposure to air.