title: Nicotine and metabolites determination in human plasma by ultra performance liquid chromatography-tandem mass spectrometry: a simple approach for solving contamination problem and clinical application.
abstract: A quantitative method using ultra performance liquid chromatography-tandem mass spectrometry is described for simultaneous determination of nicotine and its metabolites (cotinine and trans-3'- hydroxycotinine) in human plasma. Aliquots of 0.25 mL of plasma specimens were used for analysis, and 3 analytes were extracted by liquid-liquid extraction. The main problem was blank plasma contamination with environmental nicotine. Activated charcoal was used to avoid this analytical interference. For optimized chromatographic performance, a basic mobile phase consisting of 0.2% ammonia in water (mobile phase A, pH10.6) and acetonitrile (mobile phase B) was selected. The analytes were separated on a 50 mm × 2.1 mm BEH C18 column, 1.7 μm particle size, and quantified by MS/MS using multiple-reaction monitoring (MRM) in positive mode. The chromatographic separation was achieved in 3 min followed by 1.2 min of column equilibration. The calibration curves were linear in the concentration range of 10-1000 ng/mL with correlation coefficients exceeding 0.99. Within-day precisions and between-day precisions (CV, %) were <15 %. The accuracy expressed as bias was within ±15% for all analytes. The recovery values ranged from 50% to 97%. The ions used for quantification of nicotine, cotinine and 3-OH-cotinine were 166.9 > 129.7; 176.9 > 79.7; 192.9 > 79.7 m/z, respectively. The original blank sample preparation solved the problem of contamination in a cost-effective and efficient way. The validated method has been routinely used for analysis of nicotine and metabolites and determination of hydroxycotinine/cotinine metabolic ratio. This biomarker seems to be interesting at predicting response of nicotine patch replacement therapies.
authors.affiliation: Service de Pharmacologie Clinique, Faculté de Médecine, Aix-Marseille Université, 27 Bd Jean Moulin F-13385 Marseille Cedex 05, Marseille, France.
authors.forename: Natalia
authors.initials: N
authors.lastname: Liachenko
authors.affiliation: Service de Pharmacologie Clinique, Faculté de Médecine, Aix-Marseille Université, 27 Bd Jean Moulin F-13385 Marseille Cedex 05, Marseille, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Boulamery
authors.affiliation: Service de Pharmacologie Clinique, Faculté de Médecine, Aix-Marseille Université, 27 Bd Jean Moulin F-13385 Marseille Cedex 05, Marseille, France.|INSERM U912 (SESSTIM), Aix-Marseille Université, F-13006, Marseille, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Simon
mesh_terms: D000279:Administration, Cutaneous; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001711:Biotransformation; D002138:Calibration; D002606:Charcoal / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards; D003367:Cotinine / Q000031:analogs & derivatives* / Q000097:blood; D005260:Female; D006801:Humans; D059625:Liquid-Liquid Extraction; D008297:Male; D008875:Middle Aged; D009538:Nicotine / Q000008:administration & dosage / Q000097:blood*; D018722:Nicotinic Agonists / Q000008:administration & dosage / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D016540:Smoking Cessation / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000592:standards; D061485:Tobacco Use Cessation Devices; D057968:Transdermal Patch; D055815:Young Adult
chemical_list: D018722:Nicotinic Agonists; D002606:Charcoal; C001381:hydroxycotinine; D009538:Nicotine; D003367:Cotinine
keywords: UPLC/MS-MS; activated charcoal; cotinine; nicotine; trans-3′-hydroxycotinine