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  • Title: Fluorescence detection of cardenolides in reversed-phase high-performance liquid chromatography after post-column derivatization.
    Author: Belsner K, Büchele B.
    Journal: J Chromatogr B Biomed Appl; 1996 Jun 28; 682(1):95-107. PubMed ID: 8832430.
    Abstract:
    Methods for the fluorescence derivatization of cardiac glycosides with concentrated acids from TLC are adopted to HPLC for post-column derivatization. The column effluent is blended with concentrated acids in a knitted tube reactor, which enables derivatization with negligible increase in chromatographic peak width. The selectivity of the reaction is temperature-dependent and influenced by the respective acid. Reactivity increases from H3PO4-->CH3SO3H[symbol: see text]H2SO4. The conversion of digoxigenin, digitoxigenin and their digitoxosides is accelerated by Cu(II) acetate or Co(II) nitrate in H2SO4. Combined with a new two-mode, single-column solid-phase sample preparation, cardiac glycoside levels of less than 100 pg/glycoside in 1 ml plasma are detectable.
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