High-performance liquid chromatographic determination of amino acids and oligopeptides by pre-column fluorescence derivatization with 9-fluorenyl-methoxy carbonyl succinimide

A sensitive high-performance chromatographic method for the detection of amino acids and oligopeptides with pre-column fluorescence derivatization has been developed. Glycine, glycylglycine, triglycine, glutathion, glutamic acid, and cysteine were separated on a reversed-phase C-18 column with methanol-water-triethylamine as eluent; the derivatization and the chromatographic conditions were optimized. The six derivatives were eluted in 20 min with good reproducibility. The relative standard deviations (n=6) at an analytical concentration of 2 x 10(-6) M are less than 4.5%. The detection limits (signal-to-noise ratio=3) for the six derivatives are in the range of 15-46 fmol.