Optimization and validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of nicotine, cotinine, trans-3'-hydroxycotinine and norcotinine in human oral fluid.
An analytical procedure was developed and validated for the simultaneous identification and quantification of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in 0.5 mL of human oral fluid collected with the Quantisal oral fluid collection device. Solid phase extraction and liquid chromatography-tandem mass spectrometry with multiple reaction monitoring were utilized. Endogenous and exogenous interferences were extensively evaluated. Limits of quantification were empirically identified by decreasing analyte concentrations. Linearity was from 1 to 2,000 ng/mL for nicotine and norcotinine, 0.5 to 2,000 ng/mL for trans-3'-hydroxycotinine, and 0.2 to 2,000 ng/mL for cotinine. Correlation coefficients for calibration curves were >0.99 and analytes quantified within +/-13% of target at all calibrator concentrations. Suitable analytical recovery (>91%) was achieved with extraction efficiencies >56% and matrix effects <29%. This assay will be applied to the quantification of nicotine and metabolites in oral fluid in a clinical study determining the most appropriate nicotine biomarker concentrations differentiating active, passive, and environmental nicotine exposure.
Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Biomedical Research Center, 251 Bayview Boulevard Suite 200, Room 05A-721, Baltimore, MD 21224, USA. shakleyad@intra.nida.nih.gov
Diaa M
DM
Shakleya
Marilyn A
MA
Huestis
D002853:Chromatography, Liquid / Q000379:methods; D003367:Cotinine / Q000031:analogs & derivatives* / Q000737:chemistry*; D005883:Gingival Crevicular Fluid / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D009538:Nicotine / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C066333:norcotinine; C001381:hydroxycotinine; D009538:Nicotine; D003367:Cotinine