Rapid and sensitive high-performance liquid chromatographic determination of nicotine and cotinine in nonsmoker human and rat urines

J Chromatogr B Biomed Sci Appl. 1998 Apr 24;708(1-2):95-101. doi: 10.1016/s0378-4347(97)00632-4.


A simple reversed-phase high-performance liquid chromatographic method with paired-ion and UV detection has been developed for the rapid quantification of urinary nicotine and cotinine. A one-step solid-liquid extraction on Extrelut was used. Separation from endogenous substances was achieved with a decreasing flow-rate. With 20 ml of urine for extraction, the limit of quantification was 0.5 ng/ml for cotinine and 5 ng/ml for nicotine; linearity was obtained from 50 to 5000 ng/ml. The intra- and inter-day coefficients of variation were less than 9% for cotinine and 30% for nicotine. Average recoveries for cotinine were 92-100% and 47-86% for nicotine. The present method was applied to the urine analysis of smokers, nonsmoker children, and experimental animals.

MeSH terms

  • Adolescent
  • Adult
  • Animals
  • Child
  • Chromatography, High Pressure Liquid / methods*
  • Cotinine / urine*
  • Evaluation Studies as Topic
  • Female
  • Humans
  • Male
  • Nicotine / urine*
  • Rats
  • Sensitivity and Specificity
  • Smoking*
  • Time Factors
  • Tobacco Smoke Pollution


  • Tobacco Smoke Pollution
  • Nicotine
  • Cotinine