Name: Ferhaeeza Jalaidi Kalayakan Date: March 18, 2021 SCORE: ____/65______

Lab. Teacher: Prof. Evelyn Bucoy Course & Section: BS BIO 2C

ACTIVITY NO. 8
COLOR REACTION OF PROTEINS
Results: (30pts)

Test Albumin
Gluten Casein
Yielded a Positive Yielded a Positive Yielded a Positive
result in biuret test result in biuret test result in biuret test
Biuret with dark blue with purple with purple
coloration. coloration. coloration.

Intense blue color Intense blue color is Pink solution

is formed and it formed and it
Ninhydrin confirms the confirms the presence
presence of of proteins.
proteins.

A yellow A yellow precipitate Colorless solution

Xanthoproteic precipitate is is formed.
formed.
a.With HNO3

Cloudy white After 20 drops of A yellow solution

solution turned to NaOH clear neon is formed.
clear solution after yellow at 15 drops
b. with NH4OH water bath then (basic)
clear yellow
solution

Millon’s White precipitate No characteristics

 which changes to change.
brick red on
boiling.

Clear solution with Slightly yellow Colorless solution

Hopkin’s-Cole
purplish interphase

clear solution Colorless solution Murky solution

Lead Acetate turned into brown
solution

Pauly

Bromine Water

Clear red orange Clear grayish solution Saturated yellow

Sakaguchi
solution turned into red solution
solution

Conclusion: (2pts)
Proteins are made of amino acids. When amino acids are tested on, it reacts due to its amphoteric
nature and the R-group or side chain. An amphoteric means it is capable of reacting to both acids
and bases. The side chain or functional groups present in amino acid determines the intensity of
the product color. The greater number of functional groups that reacted, the more saturated its
hue. Specific tests for amino acid would express the amino acid content of a protein.

ACTIVITY NO. 8
 COLOR REACTION OF PROTEINS

Question Guide:

1. Write the positive result, the chemical group responsible for the positive and the importance of
each test. (27pts)

Test Positive Result Chemical Group Importance

The appearance of C2H5N3O2 mainly because it can detect the

bluish- violet color amount of protein in urine. Excess
Biuret confirms the of protein can lead to kidney
presence of Proteins. disease and other health-related
conditions like diabetes mellitus
and high blood pressure.
The appearance of C9H6O4 used to detect the presence of
deep blue or purple amino acids in unknown samples.
color confirms the This test is also used in solid-phase
Ninhydrin presence of proteins. peptide synthesis to monitor the
protection for amino acid analysis
of proteins. As the ninhydrin test is
quite sensitive, it is commonly
used to detect fingerprints.
the appearance of C34H24N4O122HO a method that can be used to detect
yellow color a presence of protein soluble in a
Xanthoproteic confirms the solution, using concentrated nitric
presence of proteins. acid. The test gives a positive
result in amino acids carrying
aromatic groups, especially in the
presence of tyrosine.
The appearance of HgN2O6 used for the detection of tyrosine-
brick red color containing proteins in a given
confirms the sample. The test also helps in the
Millon’s presence of proteins. differentiation of tyrosine from
other amino acids. The test is
useful in the detection of casein
protein and the protein found in
raw meat.
Hopkins-Cole  A purple ring H2SO4 is one of the color reactions used
appears between the for the detection of particular
two layers if amino acids or proteins on the
the test is positive fo basis of the formation of a specific
 r tryptophan. color. The test detects the indole
ring that is found in tryptophan
amino acid, which in turn helps in
the identification of proteins
containing tryptophan.
The Formation of Pb(C2H3O2)2 A test to detect hydrogen sulfide in
black precipitate a fluid by discoloration of a paper
indicate the presence moistened with the lead
Lead Acetate of sulfur-containing acetate solution. It is important to
amino acid. determine the presence and amount
of hydrogen sulfide because this
gas is extremely poisonous, highly
flammable, explosive and
corrosive.
A positive result is C14H14N3NaO3S  is used to detect the presence of
demonstrated by the tyrosine and histidine-containing
appearance of a red- proteins. The test also allows the
Pauly colored complex, differentiation of histidine and
indicating the tyrosine from other amino acids.
presence of histidine
and tyrosine in the
solution.
It is a qualitative test, Br2 It is a qualitative test, used to
used to identify the identify the alkene or alkane
Bromine alkene or alkane functional groups present in the
Water functional groups compound. Alkene groups react
present in the with bromine water in the dark
compound. condition and undergo an addition
reaction, to give a
decolorized solution.
Sakaguchi A positive result on C3H7NO2S a chemical test used for detecting
the Sakiguchi’s test the presence of arginine in
is demonstrated by proteins. It is named after the
the formation of red Japanese Food Scientist and
color. This indicates Organic Chemist,
the presence of an Shoyo Sakaguchi (1900–1995)
arginine or who described the test in 1925.
guanidinium The Sakaguchi reagent used in
compound. the test consists of 1-Naphthol and
a drop of sodium hypobromite.

2. What color will be produced by proline with ninhydrin test? Why? (2pts)
 amino acids, e.g., proline and hydroxyproline, react with ninhydrin to give a yellow
color.

3. Which test can be used to show the extent of hydrolysis of proteins? Why? (2pts)

The Biuret test can show the extent of hydrolysis of proteins due to
the coordination complex with the peptide bond of the protein from the CuSO4.

4. Classify proteins according to nutritional value. To which classification does albumin, gluten and
casein belong? (3pts)

Proteins are complex, organic compounds composed of many amino acids linked together
through peptide bonds and cross-linked between chains by sulfhydryl bonds, hydrogen
bonds and van der Waals forces. There is a greater diversity of chemical composition in
proteins than in any other group of biologically active compounds. The proteins in the
various animal and plant cells confer on these tissues their biological specificity.

Proteins can be classified as:

(a) Simple proteins. On hydrolysis they yield only the amino acids and occasional small
carbohydrate compounds. Examples are: albumins, globulins, glutens, albuminoids, histones and
protamine’s.

(b) Conjugated proteins. These are simple proteins combined with some non-protein material in
the body. Examples are: nucleoproteins, glycoproteins, phosphoproteins, hemoglobin and
lactoproteins.

(c) Derived proteins. These are proteins derived from simple or conjugated proteins by
physical or chemical means. Examples are: denatured proteins and peptides.