Automated determination of verapamil and norverapamil in human plasma with on-line coupling of dialysis to high-performance liquid chromatography and fluorometric detection

Journal of Chromatography. a
A CeccatoJ Crommen

Abstract

A fully automated method for the simultaneous determination of verapamil and its main metabolite norverapamil in human plasma is described. This method is based on on-line sample preparation using dialysis followed by clean-up and enrichment of the dialysate on a precolumn and subsequent HPLC analysis with fluorometric detection. All sample handling operations were performed automatically by a sample processor equipped with a robotic arm (ASTED system). The plasma samples were dialysed on a cellulose acetate membrane (cut-off: 15 kD) and the dialysate was purified and enriched on a short pre-column filled with cyanopropyl silica. Before starting dialysis, this trace enrichment column (TEC) was first conditioned with the HPLC mobile phase and then with pH 3.0 acetate buffer. 370 microliters of plasma sample spiked with the internal standard (gallopamil) were dialysed in the static-pulsed mode. The solution at the donor side was pH 3.0 acetate buffer containing Triton X-100 while the acceptor solution was made of the same acetate buffer. When dialysis was discontinued, the analytes were desorbed from the TEC by the HPLC mobile phase and transferred to the C18 analytical column by means of a switching valve. This mobile phase cons...Continue Reading

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Citations

Oct 27, 2010·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Lukáš ChytilOndřej Slanař
Nov 6, 2007·Journal of Biochemical and Biophysical Methods·Violeta IvanovaTrajce Stafilov
Jul 11, 2008·Journal of Separation Science·Imran AliAfzal Hussain
Jul 19, 2001·Journal of Chromatography. a·B M De BorbaJ Camarda
Oct 29, 2003·Human & Experimental Toxicology·A TracquiB Ludes
Oct 20, 1999·Journal of Chromatography. a·N C van de Merbel

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