Arbil polytechnic unveristy

Shaqlawa technical colege

MLT-Dipartment
Second semester

Protin test:
{ Ninhydrin & biurets test}

Prepared by : Supervisor by:

Sarkaut Muhamad. Ms:Fatima

2023–2024
Introduction
Ninhydrin and biuret tests are common chemical
assays used in biochemistry and molecular biology.
The ninhydrin test detects the presence of amino
acids or proteins by reacting with primary amines,
turning purple-blue in the process. On the other
hand, the biuret test detects the presence of peptide
bonds in proteins, forming a purple coloration when
positive. Both tests are valuable tools in biochemical
analysis for identifying and quantifying proteins and
amino acids in various samples.

Equipment & Glasswar

• Test tube

• Beaker

• Magnetic Stirrer

Chemical & Reagent

• Reagent

• Egg
 Procedure (Ninhydrin test )

1- Take 1 mL of a sample solution into a clean test tube.

2- Add a few drops of ninhydrin reagent to the test tube.
3- Place the test tube in the water bath for 5 minutes and
then allow cooling to room temperature.
4- Observe any color change.
Procedure (Biuret test )

1- Add 1 mL of the sample solution into a clean test

tube.
2- Add 1 mL of Biuret reagent to the test tube.
3- Mix well and allow the mixtures to stand for 5
minutes.
4- Observe any color change.
Discussion
The ninhydrin and biuret tests are fundamental tools in
biochemistry for identifying and characterizing proteins and
amino acids.

The ninhydrin test is highly sensitive to primary amines,

such as those found in amino acids. When exposed to
ninhydrin, these primary amines react to form a purple-blue
color, indicating the presence of proteins or amino acids.
This reaction is commonly used in chromatography to
visualize and quantify amino acids.

On the other hand, the biuret test detects the presence of

peptide bonds, which are the bonds that link amino acids
together in proteins. When proteins are treated with a
solution containing copper sulfate and sodium hydroxide
(the biuret reagent), a characteristic purple color develops if
peptide bonds are present. This color change is due to the
coordination of copper ions with the peptide bonds.

Both tests are relatively simple and widely used in

biochemical laboratories for qualitative and quantitative
analysis of proteins and amino acids. However, it's essential
to note that while the biuret test specifically detects peptide
bonds, the ninhydrin test is more general and can react with
any primary amine, not just those in peptides.
Conclusion
In conclusion, the ninhydrin and biuret tests are
indispensable tools in biochemistry for detecting and
characterizing proteins and amino acids.

The ninhydrin test provides a sensitive method for

identifying primary amines, such as those found in
amino acids, by producing a distinctive purple-blue
coloration upon reaction. This test is particularly
useful in chromatography and amino acid analysis.

On the other hand, the biuret test is specific for

detecting the presence of peptide bonds in proteins.
When treated with the biuret reagent, proteins
containing peptide bonds exhibit a characteristic
purple coloration due to the formation of a complex
with copper ions.

Both tests offer simple and reliable means of

qualitative and quantitative analysis in biochemical
research and laboratory settings, contributing
significantly to our understanding of protein structure
and composition.
 Reference

https://www.toppr.com/guides/chemistry
/ninhydrin-test/

https://byjus.com/chemistry/biuret-test/

https://en.m.wikipedia.org