Simultaneous analysis of buprenorphine, methadone, cocaine, opiates and nicotine metabolites in sweat by liquid chromatography tandem mass spectrometry.
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.
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
M
Concheiro
Diaa M
DM
Shakleya
Marilyn A
MA
Huestis
D000701:Analgesics, Opioid / Q000032:analysis*; D002047:Buprenorphine / Q000032:analysis*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D008691:Methadone / Q000032:analysis*; D009538:Nicotine / Q000032:analysis*; D013542:Sweat / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000701:Analgesics, Opioid; D002047:Buprenorphine; D009538:Nicotine; D008691:Methadone