Simultaneous determination of D- and L-amino acids in the nervous tissues of crustaceans using precolumn derivatization with (+)-1-(9-fluorenyl)ethyl chloroformate and reversed-phase ion-pair high-performance liquid chromatography.

After the derivatization of D- and L-amino acids with (+)-1-(9-fluorenyl)ethyl chloroformate, nineteen amino acids were separated into their D- and L-enantiomers and from other physiological amino compounds by reversed-phase ion-pair high-performance liquid chromatography. The separation was performed by three separate runs differing in mobile phase compositions and gradient profiles. Tyrosine, tryptophan and cysteine could not be detected because of their weak reactions with the derivatization reagent. Of seven D-amino acids found in the crustacean nervous tissues and eyes, D-alanine, D-arginine and D-aspartate were the most abundant and widely distributed.