Analysis of N′-nitrosonornicotine and its metabolites in rabbit blood with liquid chromatography/tandem mass spectrometric method

► A method to analyze NNN and its metabolites with HPLC-MS/MS was established. ► Hydroxy acid was the major metabolite of NNN in rabbit blood. ► 5′-Hydroxylation of NNN might be the main metabolic pathway of NNN in rabbit blood. ► Metabolic curves and pharmacokinetic parameters were obtained for NNN and its metabolites.