Principles of Biochemistry (2+1)

Separation of amino acid y paper chromatography

Separation of amino acid by paper chromatography

Principle

• The separation of the amino acids (solutes) is based on the liquid-liquid partitioning of amino acids by paper chromatography.
• The partitioning of amino acids take place between the water molecule (stationary phase)  adsorbed to the cellulose acetate of the paper  and the organic (mobile) phase.
• The separated amino acids can be identified by spraying ninhydrin and heating.
• The spots are marked and the R_f value calculated.
• By comparing the R_f  values the amino acids present in the sample and that of the standard amino acids, co-chromatographed, the amino acids present in the sample are identified.