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

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=310 nm, Em=380 nm). 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.