PMID: 9173684Apr 1, 1996Paper

Interspecies differences in coumarin metabolism in liver microsomes examined by capillary electrophoresis

Xenobiotica; the Fate of Foreign Compounds in Biological Systems
D P BoganM R Smyth

Abstract

1. A simple, rapid method was developed for studying xenobiotic metabolism by cytochrome P450 in liver microsome preparations. Capillary electrophoresis was used to separate the metabolite from the metabolic mixture. 2. Coumarin is metabolized to 7-hydroxycoumarin by a cytochrome P450 isoenzyme. Human, bovine, gerbil, mouse (Schofield, CO1), rat, rabbit, porcine, and cynomologus monkey microsomal preparations were investigated for coumarin metabolism by determining the content of 7-hydroxycoumarin present after metabolism. 3. Separation of 7-hydroxycoumarin from the reaction mixture was carried out in 50 mM phosphate buffer, pH 6.8, on a fused silica capillary at 25 degrees C and 15 kV. The metabolic matrix consisted of an NADPH regeneration system, 205.5 mu M coumarin, and the microsomal preparation. Standard curves were prepared in the microsomal preparation and the limit of quantification was 6.17 mu M, with a linear range from 0 to 308.5 mu M. 4. The reaction was initiated by the addition of the microsomes. An aliquot of the reaction mixture was removed at specific timed intervals over 2 h and injected directly onto a capillary electrophoresis column and the concentration of 7-hydroxycoumarin determined. The metabolism of c...Continue Reading

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Citations

May 21, 2005·Journal of Pharmaceutical and Biomedical Analysis·Jie ZhangAnn Van Schepdael
Jul 27, 1999·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·B G Lake
Oct 29, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·D LevêqueF Jehl
Aug 6, 2003·Journal of Pharmaceutical and Biomedical Analysis·Eimear M WardCraig E Lunte
Oct 9, 2008·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Ivonne M C M RietjensAns Punt
Mar 23, 2007·Electrophoresis·Jiri KonecnýZdenek Glatz
Dec 27, 2005·Journal of Chromatography. a·Pham Thi Thanh HaAnn VanSchepdael
Jul 28, 2020·Cancers·Esra Küpeli AkkolRaffaele Capasso

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