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

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 0.08–260 nmol ml−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.