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Simultaneous analysis of buprenorphine, methadone, cocaine, opiates and nicotine metabolites in sweat by liquid chromatography tandem mass spectrometry

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Abstract

A liquid chromatography tandem mass spectrometry method for buprenorphine (BUP), norbuprenorphine (NBUP), methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), cocaine, benzoylecgonine, ecgonine methyl ester (EME), morphine, codeine, 6-acetylmorphine, heroin, 6-acetylcodeine, cotinine, and trans-3′-hydroxycotinine quantification in sweat was developed and comprehensively validated. Sweat patches were mixed with 6 mL acetate buffer at pH 4.5, and supernatant extracted with Strata-XC-cartridges. Reverse-phase separation was achieved with a gradient mobile phase of 0.1% formic acid and acetonitrile in 15 min. Quantification was achieved by multiple reaction monitoring of two transitions per compound. The assay was a linear 1–1,000 ng/patch, except EME 5–1,000 ng/patch. Intra-, inter-day and total imprecision were <10.1%CV, analytical recovery 87.2–107.7%, extraction efficiency 35.3–160.9%, and process efficiency 25.5–91.7%. Ion suppression was detected for EME (−63.3%) and EDDP (−60.4%), and enhancement for NBUP (42.6%). Deuterated internal standards compensated for these effects. No carryover was detected, and all analytes were stable for 24 h at 22 °C, 72 h at 4 °C, and after three freeze/thaw cycles. The method was applied to weekly sweat patches from an opioid-dependent BUP-maintained pregnant woman; 75.0% of sweat patches were positive for BUP, 93.8% for cocaine, 37.5% for opiates, 6.3% for methadone and all for tobacco biomarkers. This method permits a fast and simultaneous quantification of 14 drugs and metabolites in sweat patches, with good selectivity and sensitivity.

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Acknowledgments

This research was supported by the Intramural Research Program of the National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), and NIDA grant RO1 DA12220. Sweat patches were generously provided by PharmChem Laboratories, Inc, Fort Worth, Texas, USA.

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Authors and Affiliations

  1. Servicio de Toxicología Forense, Dpto. Anatomía Patológica y Ciencias Forenses, Facultad de Medicina, Universidad de Santiago de Compostela, C/San Francisco s/n, 15782, Santiago de Compostela (A Coruña), Spain

    Marta Concheiro

  2. Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, 251 Bayview Boulevard, Suite 200, Room 05A721, Baltimore, MD, 21224, USA

    Marta Concheiro, Diaa M. Shakleya & Marilyn A. Huestis

  3. Chemistry and Drug Metabolism, Biomedical Research Center (BRC), National Institute on Drug Abuse, NIH, 251 Bayview Blvd., Suite 200, Room 05A721, Baltimore, MD, 21224, USA

    Marilyn A. Huestis

Authors
  1. Marta Concheiro
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  2. Diaa M. Shakleya
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  3. Marilyn A. Huestis
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Corresponding author

Correspondence to Marilyn A. Huestis.

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Published in the special issue Forensic Toxicology with Guest Editors Hans Frank T. Peters, Hans H. Maurer, and Frank Musshof.

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Concheiro, M., Shakleya, D.M. & Huestis, M.A. Simultaneous analysis of buprenorphine, methadone, cocaine, opiates and nicotine metabolites in sweat by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 400, 69–78 (2011). https://doi.org/10.1007/s00216-010-4392-2

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  • DOI: https://doi.org/10.1007/s00216-010-4392-2

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