Analysis of [3',3'-d(2)]-nicotine and [3',3'-d(2)]-cotinine by capillary liquid chromatography-electrospray tandem mass spectrometry

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Sep 15;857(1):1-8. doi: 10.1016/j.jchromb.2007.06.018. Epub 2007 Jun 29.


A selective and sensitive LC/MS/MS assay was developed for the quantification of d(2)-nicotine and d(2)-cotinine in plasma of current and past smokers administered d(2)-nicotine. After solid phase extraction and liquid-liquid extraction, HPLC separation was achieved on a capillary hydrophilic interaction chromatography phase column. The analytes were monitored by tandem mass spectrometry with electrospray positive ionization. Linear calibration curves were generated for d(2)-nicotine (0.03-6.0 ng/ml plasma) and d(2)-cotinine (0.15-25 ng/ml plasma). The lower limits of quantitation were 0.15 ng/ml and 0.25 ng/ml for d(2)-nicotine and d(2)-cotinine, respectively. The coefficient of variation was 3.7% for d(2)-nicotine and 2.5% for d(2)-cotinine. The method was applied to two ongoing studies of d(2)-nicotine metabolism in prior and current smokers. Preliminary analysis of a subset of subjects from these studies detected a significantly lower rate of nicotine conversion to cotinine by past smokers compared to current smokers.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Administration, Oral
  • Biomarkers / blood
  • Calibration
  • Capillary Electrochromatography / methods*
  • Chromatography, High Pressure Liquid / methods
  • Cotinine / analogs & derivatives
  • Cotinine / blood*
  • Cotinine / chemistry
  • Deuterium / chemistry
  • Humans
  • Injections, Intravenous
  • Molecular Structure
  • Nicotine / analogs & derivatives*
  • Nicotine / blood
  • Nicotine / chemistry
  • Radioactive Tracers
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Smoking / blood*
  • Smoking Cessation
  • Solid Phase Extraction
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Substance Abuse Detection
  • Tandem Mass Spectrometry / methods*


  • 3',3'-dideuteronicotine
  • Biomarkers
  • Radioactive Tracers
  • Nicotine
  • Deuterium
  • Cotinine