Liquid chromatographic determination of amino acids and peptides by pre-column fluorescence derivatization with N-hydroxysuccinimidyl-α-(9-acridine)-acetate

A sensitive liquid chromatographic method for the detection of amino acids and peptides with pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, triglycine, glutathione (GSH), glutamic acid, and cysteine were separated on a reversed-phase C₁₈ column. At the same time, 15 representative amino acids were also separated with a gradient program. Derivatization and the chromatographic conditions were optimized in experiments. The derivatives were eluted with a good reproducibility. The linear range of the calibration graph was 0.08-260nmolml^-1. The relative standard deviations (n=6) are 4.5%. Detection limits (signal-to-noise ratio=3) for derivatives are at the level of fmol. The methods described are suitable for the analysis of amino acids and peptides in various biological samples. Copyright (C) 1998 Elsevier Science B.V.