Micellar thin-layer chromatographic separation and identification of amino acids: Separation of L-proline from other aliphatic and aromatic amino acids

Mohammad, A.; Agrawal, V.

Journal of Planar Chromatography - Modern TLC 13(5): 365-374


Accession: 010981615

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Cationic and anionic surfactant-mediated systems have been used as mobile phases for thin-layer chromatographic separation of aliphatic and aromatic amino acids on alumina and on Li+-impregnated alumina layers. The results obtained with nonmicellar (i.e. water) and micellar (1% aqueous sodium dodecyl sulfate and cetyl trimethyl ammonium bromide) mobile phases have been compared. Addition of alcohol to micellar solutions improves the separation of amino acids. The mobility sequence of amino acids was almost identical in buffered and nonbuffered micellar mobile phases. The best TLC system for rapid separation of amino acids was plain alumina as stationary phase and 1% cetyl trimethyl ammonium bromide in water-butanol, 95 + 5, as mobile phase. With this system, L-proline was selectively separated from other aliphatic and aromatic amino acids. The lower limit of detection of amino acids has been determined, and the semiquantitative determination of DL-aspartic acid has been attempted. A linear relationship was established between amino acid RF value and the number of carbon atoms in the amino acid molecule. The proposed method is suitable for identification of L-proline, L-hydroxyproline, DL-alanine and DL-phenylalanine in spiked samples of liver, stomach, and human blood.