Determination of nicotine metabolites in urine by liquid chromatography-mass spectrometry: trends of tobacco products use in sport
This project proposes to develop and validate a method for the quantification of nicotine and its phase I metabolites (cotinine, trans-3'-hydroxycotinine, nicotine-1'-N-oxide and cotinine-N-1-oxide) in urine by LC-MS/MS. Then, a comparative assay is performed on biological samples to measure the prevalence of nicotine in sport. This study should also determine the individual patterns of consumption and allow to distinguish between the metabolism of nicotine from smoke and smokeless tobacco products.
Nicotine is the main natural alkaloid present in tobacco leaves. A wide variety of patterns of tobacco consumption exist, either smoked (cigarettes, cigars or pipes) or not smoked (chewing tobacco, snus and snuff). These multiple forms of use are consistent with various nicotine concentrations and metabolic pathways. Indeed, when tobacco is smoked, nicotine is rapidly absorbed through the lungs. On the contrary, with consumption of chewing tobacco or snus, nicotine is primarily absorbed by the oral mucosa and to a lower extent by the small intestine when saliva is swallowed.
Nicotine has a stimulating effect, increasing the pulse rate and blood pressure, but also the release of sugar in blood and the release of adrenaline. Nicotine also induces relaxing effects, reducing stress, increasing alertness and improving cognitive function. Thus, the consumption of nicotine as in smokeless tobacco products can benefit the practice of sport, without the health problems generally associated with smoke. Indeed, the use of snus and chewing tobacco is increasing, especially among winter sports such as ice hockey and skiing, but also in other sports like football or basketball and even fencing or shooting. However, only vague estimates of these nicotine consumption patterns are reported and this potential problem is underestimated, as it is not on the 2009 WADA’s (World Anti-Doping Agency) Prohibited List or in the 2009 Monitoring Program.
As a first step, this study should help to establish the prevalence of nicotine use amongst a population of athletes and to evaluate the intervariability of this consumption. Then a source inference model will be developped in order to distinguish the metabolism of nicotine between smoke and smokeless tobacco. This model will be applied to the data previously collected from athletes to determine the consumption form of each individual. This will allow sport authorities to identify the current practices and their popularity in order to consider this substance under a new anti-doping perspective.
- LC-MS/MS analysis of urine samples from athletes in competition to determine the concentrations of nicotine, cotinine, trans-3'-hydroxycotinine, nicotine-1'-N-oxide and cotinine-1-N-oxide;
- Determination of the prevalence and dose of nicotine consumed by these subjects;
- Development of a source inference model by means of statistical analysis, to distinguish the use of nicotine in smoke and smokeless tobacco;
- Application of the model to the data collected for these athletes in competition to determine the trends of consumption patterns.
Physiological effects of nicotine