title: Identification and quantification of nicotine biomarkers in human oral fluid from individuals receiving low-dose transdermal nicotine: a preliminary study.
abstract: The objective of this preliminary study was to identify and quantify potential nicotine (NIC) biomarkers in post-exposure oral fluid samples collected from 10 NIC-abstinent human participants administered 7 mg transdermal NIC using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Oral fluid samples were collected prior to NIC patch application and at 0.5 and 0.75 h after patch removal using the Quantisal() oral fluid collection device. The validated LC-MS-MS analyte panel included nicotine-Nbeta-D-glucuronide, cotinine-N-oxide, trans-3-hydroxycotinine, norcotinine, trans-nicotine-1'-N-oxide, cotinine (COT), nornicotine, NIC, anatabine, anabasine, and cotinine-N-beta-D-glucuronide. Analytes and corresponding deuterated internal standards were extracted by solid-phase extraction. NIC and COT concentrations were quantifiable in oral fluid samples collected from 6 of the 10 participants 0.5 h after patch removal and in oral fluid samples collected from 7 of the 10 participants 0.75 h after patch removal. Based on the mean NIC and COT concentrations in oral fluid and plasma for the participants with both quantifiable NIC and COT at the 0.5 and 0.75 h collection times, the oral fluid-plasma ratio was 6.4 for NIC and 3.3 for COT. An ELISA procedure was also validated and successfully applied as a screening tool for these oral fluid samples in conjunction with LC-MS-MS confirmation. An ELISA cut-off concentration of 5.0 ng/mL provided excellent sensitivity for discrimination of COT-positive post-exposure oral fluid samples collected after low-level transdermal NIC exposure and oral fluid samples collected prior to patch application.
authors.affiliation: University of Utah, Center for Human Toxicology, Department of Pharmacology and Toxicology, Salt Lake City, Utah 84108, USA. Eleanor.Miller@utah.edu
authors.forename: Eleanor I
authors.initials: EI
authors.lastname: Miller
authors.forename: Hye-Ryun K
authors.initials: HR
authors.lastname: Norris
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Rollins
authors.forename: Stephen T
authors.initials: ST
authors.lastname: Tiffany
authors.forename: Christine M
authors.initials: CM
authors.lastname: Moore
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Vincent
authors.forename: Alpana
authors.initials: A
authors.lastname: Agrawal
authors.forename: Diana G
authors.initials: DG
authors.lastname: Wilkins
mesh_terms: D000279:Administration, Cutaneous; D015415:Biomarkers / Q000032:analysis; D002853:Chromatography, Liquid; D003367:Cotinine / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D009538:Nicotine / Q000008:administration & dosage / Q000032:analysis*; D018722:Nicotinic Agonists / Q000008:administration & dosage / Q000032:analysis*; D015203:Reproducibility of Results; D012463:Saliva / Q000737:chemistry*; D052616:Solid Phase Extraction; D015813:Substance Abuse Detection / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D015415:Biomarkers; D018722:Nicotinic Agonists; D009538:Nicotine; D003367:Cotinine