Test for proteins

DISCUSSION

• Proteins are AMPHOTERIC (can act as base and

an acid)- D. Reaction with concentrated Acids
• Acts as BUFFER (neutral pH) • 1ml of filtered albumin solution, HCl,
• High MP and soluble in POLAR solvents like ionic HNO3.
substances
• Denatured in heat, addition of acids, bases or
alcohols
o Denaturation-pagkamatay/sira ng
protein
• Flocculation is a reversible process while
E. Reaction with alcohol
coagulation in albumin is irreversible
• Increasing concentration, greater
coagulation
• 10ml of 95%, 70% and 40% ethanol, 2ml
A. Reaction of heat
egg albumin
+ coagulation
➢ experts say 70% (concentration) is
actually better for disinfecting. It has
more water, which helps it to dissolve
more slowly, penetrate cells, and kill
bacteria

B. Reaction with Salts of Heavy Metals

Metals: CHROMIUM, LEAD, MERCURY, SPECIFIC TESTS

CADMIUM, ARSENIC
• Biuret Test
• Ninhydrin Test
Heavy metals cause precipitation of proteins • Xanthoproteic Test
• The more alkalinic the protein, • Hopkins Cole Test
greater chance of precipitation • Millon's Test
• 1% AgNO3, 5% HgCl2
A. Biuret Test
➢ General test for Proteins based on the
peptide bonds present.
o Reagent: 1% cupric sulfate and 10%
NaOH
(+) formation of Violet to pink complex due to the
coordination of cupric ion to basic solution

C. Reaction with Alkaloidal reagents

• 2ml of egg albumin, 1ml of saturated
picric acid, 10% tannic acid and TCA

• Place 1ml of the following solution: 1% gelatin, 1% egg

albumin, and 1% glycine
• Add 1ml of the biuret reagent (1ml of 10% NaOH and
2gtts of 1% cupric sulfate)

B. Ninhydrin Test
➢ Involves several complex reactions
between amino group and alpha Carbon
• Mix 10gtts of Hopkin's Cole Reagent to
and the ninhydrin solution.
1ml of the solution to be tested. 1%
(+: Appearance of intense blue colored solution)
albumin, 1% Casein, 1% Gelatin and 1%
Reagent: Ninhydrin reagent (0.1gm ninhydrin in 100ml
Tryptophan
of water)
• Add 10gtts of concentrated sulfuric acid
down the side of the inclined test tube.

• Mix 1ml of 0.2% ninhydrin solution with 10 gtts of egg
albumin, and alanine
• Heat for several minutes in a water bath
C. Xanthoproteic Test
➢ Positive to Tyrosine, Tryptophan and
Phenylalanine.
➢ Specific test for Amino acids with
aromatic ring
Concentrated Nitric Acid
+ appearance of yellow precipitate
E. Millon's Test
➢ Specific test for proteins containing phenol
group in the side chains such as tyrosine
+ formation of flesh to red-colored precipitate
Reagent: Millons Reagent: Dissolve 160 grams of
mercuric nitrate and 160 grams of mercurous
nitrate in 400 ml concentrate nitric acid solution.
The reagent is then made to 1000 ml by the
addition of 600ml distilled water. The formula can
be adjusted to suit the performance parameters.

• Add 5 gtts of concentrated nitric acid to 10 • Into each test tubes, place 1ml of the sample
drops of solution to be used in separate test • Add 10 gtts of Millon's Reagent, shake and boil in
tubes. Heat for several minutes in boiling water bath for 10 minutes
water bath
• Let it cool in cold water and add 10% NaOH
until the solution turns into alkaline
using litmus paper

D. Hopkins Cole Test

➢ Specific test for Trypthophan (indole-
ring containing)
Reagent: Hopkins Cole reagent (Glyoxylic
acid- It can be prepared by exposing
glacial acetic acid to sunlight for a few days
and Concentrated H2SO4)
+ Presence of purple ring