Fluorescence detection of cardenolides in reversed-phase high-performance liquid chromatography after post-column derivatization

被引：16

作者：

Belsner, K

Buchele, B

机构：

[1] Inst. of Pharmacol. of Nat. Compds., University of Ulm, D-89081 Ulm

来源：

JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS | 1996年 / 682卷 / 01期

关键词：

cardenolides; glycosides; digoxin;

D O I：

10.1016/0378-4347(96)00056-4

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 = 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.

引用

页码：95 / 107

页数：13