Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes. Issue 6 (December 2016)

• Record Type: Journal Article
• Authors: Lammers, Laureen A.; Achterbergh, Roos; Pistorius, Marcel C. M.; Bijleveld, Yuma; de Vries, Emmely M.; Boelen, Anita; Klümpen, Heinz-Josef; Romijn, Johannes A.; Mathôt, Ron A. A.
• Is Part Of: Therapeutic drug monitoring. Volume 38:Issue 6(2016:Dec.)
• Publication Date: 2016-12
• Subjects: LC-MS/MS -- cytochrome P450 -- probe -- cocktail; Pharmacokinetics -- Periodicals; Patient monitoring -- Periodicals; Drugs -- Analysis -- Periodicals; Body fluids -- Analysis -- Periodicals; Drug Therapy -- Periodicals; Monitoring, Physiologic -- Periodicals; Pharmacology -- Periodicals; 615.7
• DOI: 10.1097/FTD.0000000000000338
• Languages: English
• ISSNs: 0163-4356

Abstract

Abstract : Background: The metabolic activity of P450 enzymes in vivo can be determined using selective probe drugs. The simultaneous administration of multiple CYP-specific probe drugs is commonly known as the "cocktail approach." Disadvantages of a cocktail are large volumes of samples required for analysis and time-consuming analyses. The aim of this study was to develop and validate a simplified but sensitive method for the simultaneous quantification of 5 probe drugs [caffeine (CYP1A2), metoprolol (CYP2D6), midazolam (CYP3A4), omeprazole (CYP2C19), and S -warfarin (CYP2C9)] in a previously validated cocktail using a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method. Methods: The method entailed a single method for sample preparation that enables quick processing of the samples containing all 5 probe drugs in a small volume of blood (≥10 μL) followed by a chiral and nonchiral LC-MS/MS method. The method was validated for selectivity, specificity, resolution of racemic warfarin, linearity, accuracy, imprecision, recovery, process efficiency, ionization efficiency, and carryover effect. Results: The method showed good selectivity without matrix interferences and differentiated S - and R -warfarin enantiomers with adequate resolution (Rs = 1.55). For all analytes, the mean process efficiency was >95%, and the mean ionization efficiency was >97%. Furthermore, the accuracy was between 94.9% and 108% for all analytes, and the within- and between-run imprecision were <11.7% for the lower limit of quantification and <12.6% for the middle level and upper limit of quantification. Conclusions: The method presented here enables the simultaneous quantification of the 5 probes in a very small blood volume (≥10 μL). Furthermore, it is less time consuming than previously reported methods because it requires only 1 simple method for sample preparation followed by a nonchiral and chiral LC-MS/MS method that can be performed sequentially.