Direct determination of cotinine-N-glucuronide in urine using thermospray liquid chromatography/mass spectrometry

Biol Mass Spectrom. 1994 Feb;23(2):103-7. doi: 10.1002/bms.1200230210.


A thermospray liquid chromatographic/mass spectrometric method has been developed for direct determination of cotinine-N-glucuronide in the urine of smokers. Quantification was performed using methyl-d3-cotinine-N-glucuronide as internal standard and monitoring the protonated aglycons. Using a simple preparation, urine samples from four smokers were analyzed and the results compared favorably with those from a previously reported method that quantifies aglycon release following beta-glucuronidase treatment. Amounts of cotinine-N-glucuronide found in urine from smokers ranged from less than 0.7 to 21 nmol ml-1, indicating wide inter-individual variability in the metabolic production of this metabolite. Cotinine-N-glucuronide was found to be the second most abundant urinary nicotine metabolite. A similar method was developed for trans-3'-hydroxycotinine-N-glucuronide but this compound was not detected in smokers' urine.

MeSH terms

  • Adult
  • Chromatography, Liquid
  • Cotinine / analogs & derivatives*
  • Cotinine / urine
  • Humans
  • Male
  • Mass Spectrometry
  • Smoking / urine*


  • cotinine N-glucuronide
  • Cotinine