A high-resolution method for amino acid analysis of physiological fluids containing mixed disulfides

In this paper we report a high-resolution method and special buffers for the analysis for 45 common amino acids and related compounds and seven less-common components occasionally found in physiological fluids. Careful optimization of the chromatographic protocol resolves cysteine-penicillamine mixed disulfide, penicillamine disulfide, and dihydroxyphenylalanine from cystine and methionine; cysteine-homocysteine mixed disulfide is eluted between tyrosine and phenylalanine. Argininosuccinic acid is eluted before ethanolamine; glycerophosphoethanolamine is separated from taurine; glucosaminic acid is eluted between urea and aspartic acid. A Beckman Instruments System 6300 Amino Acid Analyzer was used. System performance is examined in terms of coefficient of variation (CV) over the concentration range of 1.0-5.0 nmol/50 microL; average CV for all components is better than 1.5%. Method performance is assessed through the analysis of normal and abnormal urine and plasma specimens.