Determination of nicotine and nicotine metabolites in urine by hydrophilic interaction chromatography-tandem mass spectrometry Potential use of smokeless tobacco products by ice hockey players

Consumption of nicotine in the form of smokeless tobacco (snus snuff chewing tobacco) or nicotine-containing medication (gum patch) may benefit sport practice Indeed use of snus seems to be a growing trend and investigating nicotine consumption amongst professional athletes is of major interest to sport authorities Thus a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the detection and quantification of nicotine and its principal metabolites cotinine trans-3-hydroxycotinine nicotine-N-oxide and cotinine-N-oxide in urine was developed Sample preparation was performed by liquid-liquid extraction followed by hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS) operated in electrospray positive ionization (ESI) mode with selective reaction monitoring (SRM) data acquisition The method was validated and calibration curves were linear over the selected concentration ranges of 10-10 000 ng/mL for nicotine cotinine trans-3-hydroxycotinine and 10-5000 ng/mL for nicotine-N-oxide and cotinine-N-oxide with calculated coefficients of determination (R-2) greater than 095 The total extraction efficiency (%) was concentration dependent and ranged between 704 and 100 4% The lower limit of quantification (LLOQ) for all analytes was 10 ng/mL Repeatability and intermediate precision were <= 9 4 and <= 9 9% respectively In order to measure the prevalence of nicotine exposure during the 2009 Ice Hockey World Championships 72 samples were collected and analyzed after the minimum of 3 months storage period and complete removal of identification means as required by the 2009 International Standards for Laboratories (ISL) Nicotine and/or metabolites were detected in every urine sample while concentration measurements indicated an exposure within the last 3 days for eight specimens out of ten Concentrations of nicotine cotinine trans-3-hydroxycotinine nicotine-N-oxide and cotinine-N-oxide were found to range between 11 and 19 750 13 and 10 475 10 and 8217 11 and 3396 and 13 and 1640 ng/mL respectively When proposing conservative concentration limits for nicotine consumption prior and/or during the games (50 ng/mL for nicotine cotinine and trans-3-hydroxycotinine and 25 ng/mL for nicotine-N-oxide and cotinine-N-oxide) about half of the hockey players were qualified as consumers These findings significantly support the likelihood of extensive smokeless nicotine consumption However since such conclusions can only be hypothesized the potential use of smokeless tobacco as a doping agent in ice hockey requires further investigation (C) 2010 Elsevier B V All rights reserved