Simultaneous Analysis of Enantiomeric Amino Acids Based on Derivatization with (S)-(+)-2-t-Butyl-2-methyl-1, 3- benzodioxole-4-carboxylic Acid and High-Performance Liquid Chromatography

Abstract

The fluorescent chiral derivatizing agent in the title, (S)-TBMB carboxylic acid, was applied for highly sensitive HPLC analysis of enantiomeric amino acids. Enantiomeric amino acids derivatized with the agent could be separated as the diastereomers on ODS (C-18) column and determined in less than 0.05 picomole amounts owing to the strong fluorescent chromophore of the reagent (Ex=310nm, Em=380nm). Three types of HPLC conditions with different pH values were proposed for the group analysis of amino acids: pH=5.7 for the simultaneous analysis of all amino acids, pH=4.6 for the selective analysis of polar amino acid derivatives and pH=6.0 for that of less polar ones.