A fast, highly efficient method for the detection of nicotine and its nine metabolites in urine was established in this paper. The quantification was achieved by using d₃-nicotine, d₃-cotinine, d₃-3-OH-cotinine, d₃-nicotine glucuronic acid, d₃-cotinine glucuronic acid and d₃-3-OH-cotinine glucuronic acid as internal standards under atmosphere pressure chemical ionization (APCI) condition of LC-MS/MS. Nineteen samples were quantified and then analyzed by discriminant analysis (DA). The pretreatment of this method was simple, time-saving and all compounds were eluted within 3.8 min. The relative standard deviation (RSD%) of nicotine and its nine metabolites were between 0.5% and 5.5% with the recovery rate ranged from 94% to 109%. The linear correlations were no less than 0.995. Smokers of different levels were significantly distinguished using DA and a strong positive correlation was found between the value of nicotine and its 9 metabolites in 24 h urine and smoke amount.

Key words: LC-MS/MS, rapid determination, urine, nicotine, metabolite, discriminant analysis