HPLC - ED determination of diclofenac and its hydroxylated metabolite as a marker of metabolic activity of CYP2C6 in rats

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Authors

JUŘICA Jan KLEPÁRNÍK Martin ZAHRADNÍKOVÁ Lucia ZENDULKA Ondřej TOMANDL Josef

Year of publication 2007
Type Article in Proceedings
Conference HPLC 2007. Abstract Book.
MU Faculty or unit

Faculty of Medicine

Citation
Web 31st International Symposium on High Performance Liquid Phase Separations and Related Techniques - HPLC 2007
Field Analytic chemistry
Keywords HPLC; electrochemical detection; diclofenac; rat; cytochrome P450; CYP2C6
Description Diclofenac belongs to the group of nonsteroidal antiinflammatory drugs. Presented method allows to determine concentration of diclofenac and its metabolite 4-hydroxydiclofenac in rat liver perfusate. The isolated rat liver was perfused with Williams medium E, the marker was administered as a bolus. The samples were collected in the 30th, 60th and 120th minute after the administration. Phenacetine was used as an internal standard (I.S.). After an addition of I.S. and pH adjustment, the analytes were extracted into diethyl ether and dried out under a gentle stream of nitrogen. The evaporated organic extract was dissolved in a mobile phase and analyzed on column Luna (2) C18 using coularray detection. The coulometric detection provides better sensitivity and limits of detection comparing to the conventional HPLC-UV methods. This method is suitable for modeling and description of possible pharmacological interactions on isoform CYP2C6.
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