Quantitative Determination of Amino Acids by Reversed-Phase High Performance Liquid Chromatography after Pre-column Derivatization

摘要

A simple, accurate and reliable high performance liquid chromatographic method was developed to determine the amino acids by pre-column derivatization method using ortho-phthaldialdehyde. The resulting derivatives are separated on reverse phase C_(18e) column by gradient elution with sodium acetate buffer and acetonitrile-water and detected by ultraviolet detection at 340 nm. The amino acids separated in 25 min with fine resolution in the standard mixture were aspartate, asparagine, serine, glycine, histidine, arginine, threonine, tryptophan, proline, tyrosine, valine, methionine, isoleucine, leucine, phenylalanine and lysine. Modification of the elution program allows concurrent resolution of peaks of valine, methionine, isoleucine and leucine. Thus, the proposed method shows the promise of wide applicability in estimation of amino acids in cereals.