High-performance liquid chromatography of N-terminal tryptophan-containing peptides with precolumn fluorescence derivatization with glyoxal

摘要: A precolumn fluorescence derivatization method combined with high-performance liquid chromatography is described for the sensitive and selective determination of N-terminal tryptophan-containing peptides. The peptides tryptophan were converted into fluorescent derivatives glyoxal in a moderately acidic medium (pH 4.5). separated on reversed-phase column isocratic elution an aqueous mobile phase composed acetonitrile, methanol phosphate buffer 6.0), subsequently detected by fluorimetry. technique provided respective oligopeptides single peaks chromatography. detection limits 55-382 fmol per 100-microliters injection volume at signal-to-noise ratio 3. also allowed facile tryptophyl fragment enzyme reaction mixture dynorphin trypsin.