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Lab 1 (Chem 131 Biochemistry) : Qualitative Tests For Amino Acids

This document provides procedures for two qualitative tests to identify amino acids: 1) Solubility tests to determine how amino acids dissolve in water, alcohol, dilute acid, and dilute base. This test is applied to glycine, tyrosine, glutamic acid, and cysteine. 2) The ninhydrin test, where ninhydrin reacts with amino acids to produce a blue color, indicating the presence of an amino acid. Safety warnings are provided for handling ninhydrin.

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Rabia Saleem
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131 views

Lab 1 (Chem 131 Biochemistry) : Qualitative Tests For Amino Acids

This document provides procedures for two qualitative tests to identify amino acids: 1) Solubility tests to determine how amino acids dissolve in water, alcohol, dilute acid, and dilute base. This test is applied to glycine, tyrosine, glutamic acid, and cysteine. 2) The ninhydrin test, where ninhydrin reacts with amino acids to produce a blue color, indicating the presence of an amino acid. Safety warnings are provided for handling ninhydrin.

Uploaded by

Rabia Saleem
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Lab 1 (Chem 131 Biochemistry)

Qualitative Tests for Amino Acids


1) Solubility Tests:
The solubility of amino acids and proteins is largely dependent on the solution pH. The structural
changes in an amino acid or protein that take place at different pH values alter the relative solubility
of the molecule. In acidic solutions, both amino and carboxylic groups are protonated. In basic
solutions, both groups are deprotonated. Amino acids are essentially soluble in water. Their
solubilities in water, dilute alkali and dilute acid vary from one compound to the other depending
on the structure of their side chains. Apply this test to glycine, tyrosine, glutamic acid and cysteine.
Procedure:
i. Note the solubility of amino acids in water and alcohol by placing a small amount in a test
tube, adding a few mL of solvent and warming if necessary.
ii. Determine the amino acid solution is acidic or basic by using a litmus paper while testing
the solubility in water.
iii. Repeat the solubility test using dilute HCl and dilute NaOH.

2) Ninhydrin Test for the Identification of Amino Acids


Ninhydrin (triketohydrindene hydrate) is a chemical used to detect ammonia or primary and
secondary amines. Amino acids also react with ninhydrin at pH=4. The reduction product obtained
from ninhydrin then reacts with NH3 and excess ninhydrin to yield a blue colored substance. This
reaction provides an extremely sensitive test for amino acids. Apply this test to any of the amino
acids you choose.
WARNING: Avoid spilling ninhydrin solutions on your skin, as the resulting stains are difficult
to remove. (Ninhydrin is the most commonly used method to detect fingerprints, as the terminal
amines or lysine residues in peptides and proteins sloughed off in fingerprints react with
ninhydrin).
Procedure: -
i. To 1 mL amino acid solution add 5 drops of 0.2% ninhydrin solution in acetone.
ii. Boil over a water bath for 2 min.
iii. Allow to cool and observe the blue color formed.
Questions: Write the reaction(s) involved in Ninhydrin Test.

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