Separation of metronidazole, its major metabolites and their conjugates using dynamically modified silica

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Metronidazole has previously been used as a probe when investigating cytochrome P450 isoenzymes and thus the pattern of metabolism of this drug has been extensively studied. However, in previous investigations the conjugates were determined by indirect methods. In this paper we present a high-performance liquid chromatographic (HPLC) system for the simultaneous determination of metronidazole, its major metabolites and their glucuronic acid conjugates in biological fluids. The separation is performed using bare silica dynamically modified with N-cetyl-N,N,N-trimethylammonium bromide contained in the mobile phase. The separation of the acidic metabolites of metronidazole is greatly improved with this system compared to other published reversed-phase HPLC systems intended for the same purpose. Glucuronides of metronidazole and its hydroxy metabolites have been synthesized in vitro using rat liver microsomes and preparative HPLC. The method developed makes it possible to determine the intact glucuronic acid conjugates of metronidazole and the hydroxy metabolite in human urine.
Original languageEnglish
JournalJournal of Chromatography A
Issue number1-2
Pages (from-to)175-184
Number of pages10
Publication statusPublished - 1995

ID: 38062337