NURBIO (G)

Activity No. 3
Title: Proteins: Color Reaction

Rating:
Name: Malaica Airisse S. Mohammad Group No. 45
1. Objective ___
2. Data, Results & Discussion___
3. Q&A ___
Date Performed & Submitted: 08/05/19- 07/12/19 4. Conclusion ___
5. N&F ___

Total: ___

I. Introduction:
Protein is the basic component of living cells and is made of carbon, hydrogen,
oxygen, nitrogen and one or more chains of amino acids. The three types of
proteins are fibrous, globular, and membrane. Chemically, protein is composed of
amino acids, which are organic compounds made of carbon, hydrogen, nitrogen,
oxygen or sulfur. Amino acids are the building blocks of proteins, and proteins are
the building blocks of muscle mass.

II. Objectives:
1. To be able to determine the color outcome of each substances.
2. To be able to determine how the color of protein reacts.

III. Date, Results & Discussion

Table 1: Biuret Test

Test Solutions Reagents Added Result Explanation
1% Albumin 10% Na OH Violet color positive
1% CuSO4
1% Casein 10% Na OH Blue color Negative
1% CuSO4
1% Glycine 10% Na OH Green color Negative
1% CuSO4
1% Peptone 10% Na OH Violet color Positive
1% CuSO4
 Urean (s) 10% Na OH Pink- violet and Negative
1% CuSO4 urea at the
bottom

Discussion: Molisch’s test, it is a sensitive test among carbohydrates. Therefore,

in the reading, all solutions shows a positive result because it has the appearance
of a violet or purple color except for the unknown sugar, it shows a negative result
because the purple or violet color is not present.

Table 2: Xanthoproteic Test

Test Solutions Reagents Added Result Explanation

1% Albumin Conc. HNO3 Yellow color Positive result
Conc. NH4OH because the
solution contains
aromatic amino
acids.
1% Casein Conc. HNO3 Yellow color Positive result
Conc. NH4OH because there has
the presence of
aromatic amino
acids.
1% Gelatin Conc. HNO3 Clear color Negative result
Conc. NH4OH because the
solution doesn’t
contain aromatic
amino acids.
1% Phenol Conc. HNO3 Yellow color Positive result
Conc. NH4OH because there has
the presence of
aromatic amino
acids.

Discussion: Fehling’s test is also a test for reducing sugars. In this test the
presence of aldehydes but not ketones is detected by reduction of the deep blue solution of
copper (II) to a red precipitate of insoluble copper oxide. Therefore in this test glucose, xylose,
fructose, lactose, maltose, starch and unknown shows a positive result therefore it is a reducing
sugar while glycogen and sucrose shows a negative result therefore it is a non-reducing sugar.
Table 3: Test for Sulfur
 Test Solutions Reagents Added Result Explanation
1% Albumin 10% NaOH Black color Positive
5%
Pb(CH3COO)2
Gelatin (s) 10% NaOH Transparent Negative
5% yellow color
Pb(CH3COO)2
Peptone (s) 10% NaOH Dark brown color Positive
5%
Pb(CH3COO)2

Discussion: Benedict’s it is used to identify the presence of reducing sugars.

Therefore, in the reading, 1% lactose, 1% glucose, 1% xylose are disaccharides and
monosaccharides that reduces sugar therefore it shows a positive result because the color violet
or purple appears in the test. While sucrose and starch are non-reducing sugars therefore it does
not react.

Table 4:Hopkins-Cole Reaction

Test Solutions Reagents Added Result Explanation
1% Albumin Conc. H2SO4 Violet ring color Positive
1% Casein Conc. H2SO4 Violet ring color Positive
1% Gelatin Conc. H2SO4 No violet ring Negative

Discussion: Barfoed’s test is used for distinguishing monosaccharides from

reducing disaccharides. Monosaccharides usually react in about 1-2 minute while the reducing
disaccharides take much longer time between 7-12 minutes to react with the reagent. As you can
see in this test, 1% fructose and 1% xylose are monosaccharides beacause because it reacted in a
few minutes while the others such as glucose, lactose, maltose, sucrose and unknown sugar are
disaccharide because it has no reaction.
Table 5: Ninhydrin Reaction
Test Solutions Reagents Added Result Explanation
0.2% Albumin 0.1% ninyhdrin No change Negative
solution
Ammonia water 0.1% ninyhdrin No change Negative
solution
0.2% Urea 0.1% ninyhdrin No change Negative
solution
0.2% Glycine 0.1% ninyhdrin Violet color Positive
solution
 Discussion: The picric acid test for carbohydrates is a very sensitive chemical test
for the presence of reducing sugars. The reducing sugars react with toxic yellow crystalline solid
to form a reddish or orange color. As you can see on the table all solutions gave a positive result
that means its a reducing sugar.

IV. Answers to Questions

1. Why do all proteins give a positive result for biuret test?
The biuret test, also known as Piotrowski's test, is a chemical test used for
detecting the presence of peptide bonds. The biuret reaction can be used to
assess the concentration of proteins because peptide bonds occur with the same
frequency per amino acid in the peptide. The intensity of the color, and hence the
absorption at 540 nm, is directly proportional to the protein concentration,
according to the Beer–Lambert law, so it gives a positive result.
2. Why does Nitric Acid stain the skin with a yellow color?
. Proteins and amino acids that contain phenyl rings form a yellow
coloredcompound when treated with concentrated nitric acid. The yellow stains on
theskin are caused due to the reaction of nitric acid with protein keratin present in
theskin. This reaction is called xanthoproteic reaction.

3. Will Methionine give a positive result for the Sulfur test? Explain
. It will not give a positive result because Sulphur in methionine is inert,it does
not readily react with NaOH to form sodium sulfide which would form
black/brown ppt. with lead acetate. Getting this ppt. is the positive response in
sulphur test for amino acids. Cysteine & cystine, the other sulphur amino acids
would answer this test. Sulphur in them readily reacts in the test.
4. What grouping in Amino Acids or Proteins is responsible for the Ninhydrin
Reaction?
An example is the ninhydrin test in which the amine functional group of α-
amino acids reacts with ninhydrin to form purple-colored
compounds. Ninhydrin is used to detect fingerprints because
it reacts with amino acids from the proteins in skin cells transferred to the
surface by the individual leaving the fingerprint.

5. What color will be produced by Proline with Ninhydrin? Why?

The color is yellow why? It’s because proline has a ring structure (secondary
amino acid). The blueish-purple result is usually associated with primary amino
acids. In these amino acids, the N is free to react with ninhydrin. However, in
proline, the N is not available for reaction as it is locked in the ring structure.
Therefore no ammonia is produced, so no blue color is presented, hence the color
will be produced is Yellow.
V. Conclusion
For all the substances that is used in chemical reactions with Protein, mostly give a
positive result because it resulted a color which is the right color when basing it as a
positive or negative result and also because of the components of each substances or
reagents that which has high concentration that gives a positive effect or you could
say its reducing sugars. But some substances did not result a positive effects or the
components of each substances did not have a positive result because of the lack of
concentration, nor its a reducing sugar.

Sources/References:
https://en.m.wikipedia.org/wiki/Biuret_test

https://www.topperlearning.com/doubts-solutions/why-does-your-skin-turns-yellow-when-nitric-
acid-falls-on-skin-oppwrnn

https://www.quora.com/Why-does-methionine-not-give-a-test-for-sulfur-containing-amino-acids

https://chem.libretexts.org/Courses/Sacramento_City_College/SCC%3A_Chem_309_-
_General%2C_Organic_and_Biochemistry_(Bennett)/Text/16%3A_Proteins_and_Enzymes/16.02%3
A_Reactions_of_Amino_Acids

https://biochembro.wordpress.com/2013/02/18/yellow-result-from-ninhydrin-test/