High-performance liquid chromatographic assay for human liver microsomal omeprazole metabolism

Tommy Andersson, Per Olof Lagerstrøm, John O. Miners, Maurice E. Veronese, Lars Weidolf, Donald J. Birkett

    Research output: Contribution to journalArticle

    22 Citations (Scopus)

    Abstract

    Assays for the measurement of omeprazole metabolites in plasma and urine have been reported, but when applied to the determination of omeprazole metabolites formed by human liver microsomal incubations there were obvious limitations in sensitivity. The present high-performance liquid chromatographic (HPLC) assay, which comprises extraction, evaporation and reconstitution, is several-fold more sensitive with a limit of detection of approximately 2 pmol (2 nM in incubate) for omeprazole sulphone and 25 pmol (25 nM in incubate) for hydroxyomeprazole. Extraction efficiency is essentially quantitative and is highly reproducible (coefficient of variation = 2.1% for both metabolites). The assay is linear over a wide range of concentrations and the formation of the metabolites is linear with respect to both time (to 15 min) and protein concentration (to 1.5 mg/ml). Two minor metabolites, one of which was identified tentatively as 5-O-desmethylomeprazole, were also formed by human liver microsomes and could be determined by this method. Preliminary studies of the formation of omeprazole sulphone and hydroxyomeprazole showed that the formation kinetics in human liver microsomes were biphasic for both metabolites, suggesting that at least two different cytochrome P450 isoforms are involved in their formation.

    Original languageEnglish
    Pages (from-to)291-297
    Number of pages7
    JournalJournal of Chromatography B: Biomedical Sciences and Applications
    Volume619
    Issue number2
    DOIs
    Publication statusPublished - 22 Sep 1993

    Fingerprint Dive into the research topics of 'High-performance liquid chromatographic assay for human liver microsomal omeprazole metabolism'. Together they form a unique fingerprint.

    Cite this