Characterization by liquid chromatography-nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry of two coupled oxidative-conjugative metabolic pathways for 7-ethoxycoumarin in human liver microsomes treated with alamethicin.
作者： Michael B. Fisher , David Jackson , Andreas Kaerner , Steven A. Wrighton , Anthony G. Borel
DOI: 10.1124/DMD.30.3.270
关键词: Alamethicin 、 Metabolite 、 Liquid chromatography–mass spectrometry 、 Glucuronidation 、 Perforation (oil well) 、 High-performance liquid chromatography 、 Biochemistry 、 Chromatography 、 Glucuronide 、 Chemistry 、 Microsome
摘要: The microsomal metabolism of 7-ethoxycoumarin (7-EC) was investigated using liquid chromatography (LC)-NMR and chromatography-mass spectrometry (LC-MS) to characterize the coupling oxidative-conjugative events. Within microsomes, cytochromes P450 (P450s) UDP-glucuronosyltransferases (UGTs) are spatially disparate, each having surface luminal localization, respectively. To optimize cofactor substrate transit UGT without compromising activity, pore-forming peptide alamethicin used for perforation. Aqueous extracts incubations containing NADPH UDP-glucuronic acid were injected LC-NMR LC-MS analysis. analytical complementarity permitted identification four metabolites (M1 M4). M1 M2 novel 7-EC, consistent with 3-hydroxylation subsequent glucuronidation, Metabolites M3 M4 7-hydroxycoumarin (7-HC) 7-HC glucuronide, Viewed collectively, these results illustrate utility in examination coupled synergy metabolite identification.
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