1. Simultaneous LC-MS/MS quantification of P-glycoprotein and cytochrome P450 probe substrates and their metabolites in DBS and plasma
- Author
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Marija Bosilkovska, Julien Déglon, Caroline Samer, Bernhard Walder, Jules Desmeules, Christian Staub, and Youssef Daali
- Subjects
Male ,CYP2B6 ,Clinical Biochemistry ,Flurbiprofen ,Pharmacology ,Chromatography, High Pressure Liquid/standards ,Dextromethorphan ,Midazolam/blood/metabolism/pharmacokinetics ,Analytical Chemistry ,Substrate Specificity ,Terfenadine/analogs & derivatives/blood/metabolism/pharmacokinetics ,Cytochrome P-450 Enzyme System ,Tandem Mass Spectrometry ,Caffeine/blood/metabolism/pharmacokinetics ,Sample preparation ,P-Glycoprotein/metabolism ,Pharmaceutical Preparations/blood/metabolism ,General Pharmacology, Toxicology and Pharmaceutics ,Chromatography, High Pressure Liquid ,ddc:617 ,Chemistry ,Venous Plasma ,General Medicine ,Medical Laboratory Technology ,Tandem Mass Spectrometry/standards ,Pharmaceutical Preparations ,Calibration ,Terfenadine ,Omeprazole ,medicine.drug ,Half-Life ,Analyte ,Cytochrome P-450 Enzyme System/metabolism ,Midazolam ,In vivo ,Caffeine ,medicine ,Animals ,Dextromethorphan/blood/metabolism/pharmacokinetics ,Bupropion/blood/metabolism/pharmacokinetics ,ATP Binding Cassette Transporter, Subfamily B, Member 1 ,Bupropion ,Chromatography ,ddc:614.1 ,Flurbiprofen/blood/metabolism/pharmacokinetics ,CYP1A2 ,Omeprazole/blood/metabolism/pharmacokinetics ,Dried Blood Spot Testing - Abstract
Background: An LC–MS/MS method has been developed for the simultaneous quantification of P-glycoprotein (P-gp) and cytochrome P450 (CYP) probe substrates and their Phase I metabolites in DBS and plasma. P-gp (fexofenadine) and CYP-specific substrates (caffeine for CYP1A2, bupropion for CYP2B6, flurbiprofen for CYP2C9, omeprazole for CYP2C19, dextromethorphan for CYP2D6 and midazolam for CYP3A4) and their metabolites were extracted from DBS (10 µl) using methanol. Analytes were separated on a reversed-phase LC column followed by SRM detection within a 6 min run time. Results: The method was fully validated over the expected clinical concentration range for all substances tested, in both DBS and plasma. The method has been successfully applied to a PK study where healthy male volunteers received a low dose cocktail of the here described P-gp and CYP probes. Good correlation was observed between capillary DBS and venous plasma drug concentrations. Conclusion: Due to its low-invasiveness, simple sample collection and minimal sample preparation, DBS represents a suitable method to simultaneously monitor in vivo activities of P-gp and CYP.
- Published
- 2014