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  • Title: On-line post-column fluorescence derivatization of arginine-containing peptides in high-performance liquid chromatography.
    Author: Ohno M, Kai M, Ohkura Y.
    Journal: J Chromatogr; 1987 Apr 17; 392():309-16. PubMed ID: 3597579.
    Abstract:
    A selective detection system based on on-line post-column fluorescence derivatization is described for the analysis of arginine-containing peptides by high-performance liquid chromatography. The peptides are automatically converted into fluorescent derivatives with benzoin, a fluorogenic reagent for guanidino compounds, after separation on a reversed-phase column (TSKgel ODS-120T) and detection in an ultraviolet absorption detector. The system permits fluorescence detection at 435 nm emission with irradiation at 325 nm for arginine-containing peptides in as little as picomole amounts. The chromatogram obtained with fluorescence detection only shows peaks corresponding to arginine-containing peptides. The facile detection of arginine-containing fragments in the tryptic digest of beta-melanocyte stimulating hormone as a model compound could be achieved by comparison with a chromatogram obtained with ultraviolet absorption detection at 215 nm.
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