Rapid determination of five probe drugs and their metabolites in human plasma and urine by liquid chromatography/tandem mass spectrometry: application to cytochrome P450 phenotyping studies
Article first published online: 4 NOV 2004
Copyright © 2004 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 18, Issue 23, pages 2921–2933, 15 December 2004
How to Cite
Yin, O. Q. P., Lam, S. S. L., Lo, C. M. Y. and Chow, M. S. S. (2004), Rapid determination of five probe drugs and their metabolites in human plasma and urine by liquid chromatography/tandem mass spectrometry: application to cytochrome P450 phenotyping studies. Rapid Commun. Mass Spectrom., 18: 2921–2933. doi: 10.1002/rcm.1704
- Issue published online: 4 NOV 2004
- Article first published online: 4 NOV 2004
- Manuscript Revised: 1 OCT 2004
- Manuscript Accepted: 1 OCT 2004
- Manuscript Received: 5 AUG 2004
- Innovation and Technology Commission of the Government of Hong Kong SAR. Grant Number: ITS/174/00
- Research Grants Council of Hong Kong SAR. Grant Number: CUHK4180/02M
A liquid chromatography/mass spectrometry method, for rapid determination of five cytochrome P450 (CYP) probe drugs and their relevant metabolites in human plasma and urine, is described. The five specific probe substrates/metabolites, caffeine/paraxanthine (CYP1A2), tolbutamide/4-hydroxytolbutamide/carboxytolbutamide (CYP2C9), omeprazole/5-hydroxyomeprazole (CYP2C19), debrisoquine/5-hydroxydebrisoquine (CYP2D6) and midazolam/1′-hydroxymidazolam (CYP3A), together with the internal standards (phenacetin and paracetamol), in plasma and urine, were extracted using solid-phase extraction. The chromatography was performed using a C18 column with an isocratic mobile phase consisting of acetonitrile and 0.1% formic acid in water (70:30). The triple-quadrupole mass spectrometer was operated in both positive and negative modes, and multiple reaction monitoring was used for quantification. The method was validated over the concentration ranges 0.05–5 μg/mL for caffeine and paraxanthine, 0.02–2 μg/mL for tolbutamide, 0.1–20 μg/mL for 4-hydroxytolbutamide, carboxytolbutamide, debrisoquine and 5-hydroxydebrisoquine, 5–2500 ng/mL for omeprazole and 5-hydroxyomeprazole, and 1–100 ng/mL for midazolam and 1′-hydroxymidazolam. The intra- and inter-day precision were 0.3–13.7% and 1.9–14.3%, respectively, and the accuracy ranged from 93.5–107.2%. The lower limit of quantification varied between 1 and 100 ng/mL. The present method provides a robust, fast and sensitive analytical tool for the five-probe drug cocktail, and has been successfully applied to a clinical phenotyping study in 16 subjects. Copyright © 2004 John Wiley & Sons, Ltd.