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

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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.

Workflow overwiew of cocktail CYP-phenotyping study

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Acknowledgments

The authors thankfully acknowledge Ms Pirjo Hänninen (School of Pharmacy, University of Eastern Finland) for the excellent technical assistance, Dr. Seppo Auriola for helpful discussions and advice regarding the method validation procedure and M.Sc.Taisiya Bezhaeva for her assistance in preparation of the graphical abstract.

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Authors and Affiliations

  1. School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O. Box 1627, 70211, Kuopio, Finland

    Elena Puris, Markku Pasanen, Mikko Gynther, Merja R. Häkkinen, Tapani Keränen, Paavo Honkakoski, Hannu Raunio & Aleksanteri Petsalo

  2. Institute of Public Health and Clinical Nutrition, University of Eastern Finland, P.O. Box 1627, 70211, Kuopio, Finland

    Jussi Pihlajamäki

  3. Department of Clinical Nutrition and Obesity Center, Kuopio University Hospital, P.O. Box 1627, 70211, Kuopio, Finland

    Jussi Pihlajamäki

  4. National Institute for Health and Welfare, P.O. Box 30, 00271, Helsinki, Finland

    Tapani Keränen

Authors
  1. Elena Puris
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  2. Markku Pasanen
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  4. Merja R. Häkkinen
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  5. Jussi Pihlajamäki
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  9. Aleksanteri Petsalo
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Corresponding author

Correspondence to Elena Puris.

Ethics declarations

The pharmacokinetic study was approved by the Research Ethics Committee of North Savo District Hospital No: 27/2010 and was conducted in accordance with the Declaration of Helsinki. All healthy volunteers gave the informed consent before any study procedures were conducted.

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The authors declare that they have no conflict of interest.

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Puris, E., Pasanen, M., Gynther, M. et al. 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 409, 251–268 (2017). https://doi.org/10.1007/s00216-016-9994-x

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  • DOI: https://doi.org/10.1007/s00216-016-9994-x

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