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• W1981034810 abstract "A simplified, rapid, and selective liquid chromatography–tandem mass spectrometry method for the determination of the activities of cytochrome P450 (CYP) enzymes and UDP-glucuronosyltransferases (UGTs) in two separate settings was developed and successfully applied to 8 CYP isoenzymes and UGT2B7 enzyme activities in rat liver microsomes. The triple-quadrupole mass spectrometric detection was operated in positive mode for the probe metabolites: CYP1A2 (resorufin), CYP2B6 (hydroxybupropion), CYP2C19 (5-hydroxyomeprazole), CYP2D6 (dextrophan), CYP3A4 (6β-hydroxytestosterone), and UGT2B7 (morphine-3-glucuronide); also in negative mode for CYP2C9 (4-hydroxytolbutamide), CYP2E1 (6-hydroxychloroxazone), and CYP4A (hydroxylauric acid). The metabolic reactions were terminated with acetonitrile, containing metoprolol and acetaminophen as the internal standard for positive and negative ion electrospray ionization, respectively. The method was validated over the concentration range of 25–2500 ng/mL for 5-hydroxyomeprazole, dextrophan, hydroxylauric acid, and morphine-3-glucuronide; 5–500 ng/mL for resorufin; 3–300 ng/mL for hydroxybupropion; 10–1000 ng/mL for 4-hydroxytolbutamide; 40–4000 ng/mL for 6-hydroxychloroxazone; and 63–6300 ng/mL for 6β-hydroxytestosterone. All of the extraction recoveries of these analytes were greater than 85%, except for hydroxylauric acid at mid-concentration with a recovery of 83.2%±3.2%. The matrix effects were between 85.8% and 119.9%; the respective within- and between-run precisions were 0.9–12.0% and 2.0–13.9%; and the within- and between-run accuracy levels were 0.6–17.2% and 0.1–15.1%, respectively, for all these analytes. All of the analytes were stable during the assay and storage in the liver microsomes of Sprague-Dawley rats. The measurement activity of multiple enzymes was feasible using a cocktail approach. This method proved to be a robust, fast, accurate, specific and sensitive assay, and was successfully used to investigate in vivo enzyme activities of 8 major CYP isoenzymes and UGT2B7 in Sprague-Dawley rats with fatty livers. By the end of the eighth week, the CD-fed induced fatty liver rats showed a significant decrease in the activities of CYP1A2 and UGT2B7 as compared to the standard diet group." @default.
• W1981034810 created "2016-06-24" @default.
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• W1981034810 date "2013-03-01" @default.
• W1981034810 modified "2023-10-16" @default.
• W1981034810 title "Validated liquid chromatography–tandem mass spectrometry method for determination of totally nine probe metabolites of cytochrome P450 enzymes and UDP-glucuronosyltransferases" @default.
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• W1981034810 doi "https://doi.org/10.1016/j.talanta.2012.12.023" @default.
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