title: Simultaneous determination of nicotine and cotinine in serum using high-performance liquid chromatography with fluorometric detection and postcolumn UV-photoirradiation system.
abstract: A simple and rapid method for the simultaneous determination of serum nicotine and cotinine using high-performance liquid chromatography (HPLC)-fluorometric detection with a postcolumn ultraviolet-photoirradiation system was developed. Analytes were extracted from alkalinized human serum via liquid-liquid extraction using chloroform. The organic phase was back-extracted with the acidified aqueous phase, and the analytes were directly injected into an ion-pair reversed-phase HPLC system. 6-Aminoquinoline was used as an internal standard. Nicotine, cotinine, and 6-aminoquinoline were separated within 14min. The extraction efficiency of nicotine and cotinine was greater than 91%. The linear range was 0.30-1000ng for nicotine and 0.06-1000ng for cotinine. In serum samples from smokers, the concentrations of nicotine and cotinine were 8-15ng/mL and 156-372ng/mL, respectively.
authors.affiliation: Laboratory of Bioanalytical Chemistry, Faculty of Pharma Sciences, Teikyo University, 2-11-1 Kaga, Itabashi, Tokyo 173-8605, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Yasuda
authors.forename: Tatsuhiro
authors.initials: T
authors.lastname: Ota
authors.forename: Atsushi
authors.initials: A
authors.lastname: Morikawa
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Mawatari
authors.forename: Tomoko
authors.initials: T
authors.lastname: Fukuuchi
authors.forename: Noriko
authors.initials: N
authors.lastname: Yamaoka
authors.forename: Kiyoko
authors.initials: K
authors.lastname: Kaneko
authors.forename: Kazuya
authors.initials: K
authors.lastname: Nakagomi
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003367:Cotinine / Q000097:blood*; D005470:Fluorometry / Q000379:methods*; D006801:Humans; D008297:Male; D009538:Nicotine / Q000097:blood*; D012907:Smoking / Q000097:blood; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D009538:Nicotine; D003367:Cotinine
keywords: Cotinine; Fluorescence detection; HPLC; Liquid–liquid extraction; Nicotine; UV-photoirradiation