Page 73 - Practical book PC406-MNU-2025

MANSOURA NATIONAL UNIVERSIY
PHARM D- CLINICAL PHARMACY LEVEL II INSTRUNENTAL ANALYSIS (PC 406)

Separation and identification of amino acids

Sample Unknown mixture of amino acids

Mobile phase methyl ethyl ketone: propionic acid : Water

75 : 25 : 30

Detection 1% Ninhydrin

This method is used for identification of unknown mixture of amino acid by
applying small spots of amino acid mixture and pure amino acids .and observe

separated spots from the mixture and compare them with spots of pure amino
acids.

The amino acid mixture may be

1) Synthetic mixture of amino acids.

2) Orange juice. (to identify type of amino acids present in orange)

3) Lemon juice. (to identify type of amino acids present in lemon)

N.B:

1) We must use lead pencil, instead of a pen, to mark a chromatography paper.

2) We should avoid touching the surface of the paper to be used for amino-acid

chromatography.

3) Amino acid detected by spraying ninhydrin on the filter paper because they are

colorless but give certain color with ninhydrin.

4) Amino acid can be separated by another mobile phase:

(BAW): Butan-1-ol : Acetic acid : Water

60 : 15 : 25

45 | P a g e S e c o n d s e m e s t e r 2 0 2 4 - 2 0 2 5