title: Simultaneous quantification of nicotine, cotinine, trans-3'-hydroxycotinine, norcotinine and mecamylamine in human urine by liquid chromatography-tandem mass spectrometry.
abstract: BACKGROUND
  Mecamylamine is a nicotine antagonist under investigation in combination with nicotine replacement for smoking treatment.
  
  
  METHODS
  A simple, rapid and reliable liquid chromatography tandem mass spectrometry (LCMSMS) method was developed and validated for quantifying nicotine, cotinine, trans-3'-hydroxycotinine, norcotinine and mecamylamine in human urine. Chromatography was performed on a Synergi PolarRP column with a gradient of 0.1% formic acid and 0.1% formic acid in acetonitrile at 0.25 ml/min with an 8-min total runtime. Analytes were monitored by positive mode electrospray ionization and multiple reaction monitoring mass spectrometry.
  
  
  RESULTS
  Linear dynamic ranges were 1-500 ng/ml for nicotine and norcotinine, 0.5-500 ng/ml for trans-3'-hydroxycotinine, 0.2-500 ng/ml for cotinine, and 0.1-100 ng/ml for mecamylamine; correlation coefficients were consistently greater than 0.99, and all calibrator concentrations were within 20% of target. Extensive endogenous and exogenous interferences were evaluated. At 3 concentrations spanning the linear dynamic range of the assay, mean extraction efficiencies from urine were 55.1-109.1% with analytical recovery (bias) 82.0-118.7% and total imprecision of 0.7-9.1%. Analytes were stable for 24h at room temperature, 72 h at 4 °C, 72 h in autosampler at 15 °C and after three freeze/thaw cycles.
  
  
  CONCLUSIONS
  This method is useful for monitoring mecamylamine, nicotine and nicotine metabolites in smoking cessation and other clinical nicotine research.
authors.affiliation: Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Baltimore, MD 21224, USA.
authors.forename: Karl B
authors.initials: KB
authors.lastname: Scheidweiler
authors.forename: Diaa M
authors.initials: DM
authors.lastname: Shakleya
authors.forename: Marilyn A
authors.initials: MA
authors.lastname: Huestis
mesh_terms: D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D003367:Cotinine / Q000031:analogs & derivatives* / Q000652:urine*; D006801:Humans; D008464:Mecamylamine / Q000652:urine*; D009538:Nicotine / Q000652:urine*; D012680:Sensitivity and Specificity; D012907:Smoking / Q000652:urine; D016540:Smoking Cessation; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C066333:norcotinine; C001381:hydroxycotinine; D008464:Mecamylamine; D009538:Nicotine; D003367:Cotinine