A liquid chromatography-tandem mass spectrometry analysis of nine cytochrome P450 probe drugs and their corresponding metabolites in human serum and urine

  • Anal Bioanal Chem. 2017 Jan;409(1):251-268. doi: 10.1007/s00216-016-9994-x.
Elena Puris  1 ,  Markku Pasanen  2 ,  Mikko Gynther  2 ,  Merja R Häkkinen  2 ,  Jussi Pihlajamäki  3  4 ,  Tapani Keränen  2  5 ,  Paavo Honkakoski  2 ,  Hannu Raunio  2 ,  Aleksanteri Petsalo  2
Affiliations
  • 1. School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211, Kuopio, Finland. [email protected].
  • 2. School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211, Kuopio, Finland.
  • 3. Institute of Public Health and Clinical Nutrition, University of Eastern Finland, P.O. Box 1627, 70211, Kuopio, Finland.
  • 4. Department of Clinical Nutrition and Obesity Center, Kuopio University Hospital, P.O. Box 1627, 70211, Kuopio, Finland.
  • 5. National Institute for Health and Welfare, P.O. Box 30, 00271, Helsinki, Finland.
PMID: 27734142 DOI: 10.1007/s00216-016-9994-x
Abstract

Cocktail phenotyping using specific probe drugs for Cytochrome P450 (CYP) Enzymes provides information on the real-time activity of multiple CYPs. We investigated different sample preparation techniques and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with simple protein precipitation for the analysis of nine CYP probe drugs and their metabolites in human serum and urine. Specific CYP probe drugs (melatonin, CYP1A2; nicotine, CYP2A6; bupropion, CYP2B6; repaglinide, CYP2C8; losartan, CYP2C9; omeprazole, CYP2C19 and CYP3A4; dextromethorphan, CYP2D6; chlorzoxazone, CYP2E; midazolam, CYP3A4) and their main metabolites, with the exception of 3'-hydroxyrepaglinide, were quantified in human serum and urine using the developed LC-MS/MS method. The analytical method was fully validated showing high selectivity, linearity, acceptable accuracy (85-115 %) and precision (2-19 %) and applied to a pharmacokinetic study in four healthy volunteers after oral administration of drugs given as a cocktail. All probe drugs and their metabolites (totally 19 analytes) were detected and quantified from human serum and urine over the time range of 1 to 6 h after oral administration. Therefore, the proposed method is applicable for drug interaction and CYP phenotyping studies utilizing a cocktail approach. Graphical Abstract Workflow overwiew of cocktail CYP-phenotyping study.

Keywords
CYP; Cocktail approach; In vivo; LC-MS/MS; Pharmacokinetics; Sample preparation.
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