A one-year monitoring of nicotine use in sport: Frontier between potential performance enhancement and addiction issues

https://doi.org/10.1016/j.forsciint.2011.05.026Get rights and content

Abstract

Tobacco consumption is a global epidemic responsible for a vast burden of disease. With pharmacological properties sought-after by consumers and responsible for addiction issues, nicotine is the main reason of this phenomenon. Accordingly, smokeless tobacco products are of growing popularity in sport owing to potential performance enhancing properties and absence of adverse effects on the respiratory system. Nevertheless, nicotine does not appear on the 2011 World Anti-Doping Agency (WADA) Prohibited List or Monitoring Program by lack of a comprehensive large-scale prevalence survey. Thus, this work describes a one-year monitoring study on urine specimens from professional athletes of different disciplines covering 2010 and 2011. A method for the detection and quantification of nicotine, its major metabolites (cotinine, trans-3-hydroxycotinine, nicotine-N′-oxide and cotinine-N-oxide) and minor tobacco alkaloids (anabasine, anatabine and nornicotine) was developed, relying on ultra-high pressure liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-TQ-MS/MS). A simple and fast dilute-and-shoot sample treatment was performed, followed by hydrophilic interaction chromatography–tandem mass spectrometry (HILIC-MS/MS) operated in positive electrospray ionization (ESI) mode with multiple reaction monitoring (MRM) data acquisition. After method validation, assessing the prevalence of nicotine consumption in sport involved analysis of 2185 urine samples, accounting for 43 different sports. Concentrations distribution of major nicotine metabolites, minor nicotine metabolites and tobacco alkaloids ranged from 10 (LLOQ) to 32,223, 6670 and 538 ng/mL, respectively. Compounds of interest were detected in trace levels in 23.0% of urine specimens, with concentration levels corresponding to an exposure within the last three days for 18.3% of samples. Likewise, hypothesizing conservative concentration limits for active nicotine consumption prior and/or during sport practice (50 ng/mL for nicotine, cotinine and trans-3-hydroxycotinine and 25 ng/mL for nicotine-N′-oxide, cotinine-N-oxide, anabasine, anatabine and nornicotine) revealed a prevalence of 15.3% amongst athletes. While this number may appear lower than the worldwide smoking prevalence of around 25%, focusing the study on selected sports highlighted more alarming findings. Indeed, active nicotine consumption in ice hockey, skiing, biathlon, bobsleigh, skating, football, basketball, volleyball, rugby, American football, wrestling and gymnastics was found to range between 19.0 and 55.6%. Therefore, considering the adverse effects of smoking on the respiratory tract and numerous health threats detrimental to sport practice at top level, likelihood of smokeless tobacco consumption for performance enhancement is greatly supported.

Introduction

Origins of tobacco consumption date back to South American Aztec shamanic rites. Little could predict this ancient tradition would turn into a world epidemic due to nicotine, the principal alkaloid found in tobacco leaves where it acts as a natural insecticide.

Indeed, this molecule exhibits a variety of pharmacological properties sought-after by consumers and responsible for persistent addiction issues. Amongst the numerous neurotransmitters released in the central nervous system by stimulation of nicotinic cholinergic receptors, dopamine is associated with rewarding experiences [1]. Promotion of related positive reinforcing effects results in vigilance and cognitive function enhancement together with relaxation, reduced stress, mood modulation and lower body weight [2], [3]. Interestingly, nicotine also triggers a significant increase of pulse rate, blood pressure, blood sugar and epinephrine release owing to simultaneous stimulant and relaxant properties [4], [5]. As a consequence, nicotine addiction develops through repeated exposure to experience positive reinforcing effects with relief of withdrawal symptoms [1].

Despite public knowledge on toxicity and carcinogenic properties of tobacco smoke components, worldwide consumption is responsible for over 5 million deaths each year, a number expected to grow up to 8.3 million by 2030 [6]. Noteworthy, tobacco use extensively contributes to the burden of disease from the four leading causes of death, including ischaemic heart disease, cerebrovascular disease, infections of the lower respiratory system and chronic obstructive pulmonary disease [7]. In addition, trachea, bronchus and lung cancers mortality is an additional major health issue significantly correlated with tobacco consumption.

Therefore, treatment of nicotine addiction is a global concern which is considered from different perspectives, supporting either smoke cessation programs or preventive and harm-reducing oriented smoke regulation policies. Indeed, current strategies focus on different aspects of the problem, including various approaches such as medical advice to pharmacotherapy, or tax increases to implementation of smoke-free environments and distribution of educational material on tobacco consumption [6], [8]. At present, prohibition of smoking in public places is growing popular worldwide, proving efficient in the intent of reducing both first- and second-hand exposure while helping smokers to quit. As a consequence, the tobacco industry is in the need of marketing a diversified range of products to bypass such a restrictive legislation which is obviously detrimental to the business. Accordingly, a solid attempt to advertise smokeless tobacco products, in particular snus, is observed throughout Europe and North America. However, equivalent addictive properties have been observed when comparing smoked and smokeless nicotine, which is a serious promoting factor for smoking initiation [9]. In addition, despite avoiding health issues associated to tobacco smoke, consumption of smokeless tobacco may be responsible for oral, esophageal and pancreatic cancers due to the presence of over 28 carcinogenic constituents, but also heart diseases and lesions of the oral tissues [10], [11].

Therefore, smokeless tobacco is a very attractive drug from a doping perspective, considering the performance enhancement pharmacological properties of nicotine and the absence of direct adverse effects on the respiratory tract [12]. Actually, recent observations on nicotine consumption depict this phenomenon as a popular trend in winter sports, regardless of the level of competition [13], [14], [15]. In particular, a recent study on the 2009 Ice Hockey World Championships brought alarming findings as active nicotine consumption before or/and during the games was highlighted for about half of the athletes [16]. Nevertheless, nicotine still does not appear on the 2011 World Anti-Doping Agency (WADA) Prohibited List or Monitoring Program, despite satisfying the three inclusion criteria [17], [18]. Indeed, this substance may enhance sport performance, represents a potential health threat for the athlete and may also alter the spirit of sport due to the negative image associated with tobacco consumption. However, prevalence of nicotine consumption in sport still suffers from a lack of large-scale comprehensive survey.

Thus, this work describes a one-year monitoring study on nicotine use in a broad range of sports between 2010 and 2011. Accordingly, presence of nicotine, its four main metabolites (cotinine, trans-3-hydroxycotinine, nicotine-N′-oxide and cotinine-N-oxide) and three minor tobacco alkaloids (anatabine, anabasine and nornicotine) (Fig. 1) was investigated in over 2000 urine collected during regular doping protocols, using a dilute-and-shoot sample preparation followed by ultra-high pressure liquid chromatography-triple quadrupole mass spectrometry (UHPLC-TQ-MS/MS) in hydrophilic interaction chromatography (HILIC) mode.

Section snippets

Reagents and chemicals

(S)-Nicotine (≥99%) and (S)-cotinine (98%) were purchased from Sigma–Aldrich (Buchs, Switzerland). trans-3-Hydroxycotinine (99.9%), (R/S)-nicotine-N′-oxide (98%), (S)-cotinine-N-oxide (98%), (R/S)-anatabine (98%), (R/S)-anabasine (98%), (S)-nornicotine (98%) and (R/S)-d4-anatabine (98%) were obtained from Toronto Research Chemicals (Toronto, Canada). (S)-d4-Nicotine (98.8%), (R/S)-d3-cotinine (99%) and d3-trans-3-hydroxycotinine (98%) were supplied by LGC Promochem (Molsheim, France).

UHPLC–MS/MS analysis

Compounds of interest, including nicotine and phase I metabolites along with minor tobacco alkaloids, were selected to highlight recent consumption of tobacco but also to gather comprehensive information on metabolism patterns to help distinguish between smoke and smokeless consumption in a future retrospective study. While phase II glucuronide conjugates of some metabolites may be excreted predominantly in urine, these compounds were not investigated. Indeed, the primary focus was on

Conclusion

As a response to smoking prohibition policies flourishing throughout the world, the tobacco industry initiated a strong marketing process of various smokeless products, in particular snus. With progression of consumption in society, smokeless tobacco is drawing attention in the sport community due to performance enhancement pharmacological properties of nicotine free of smoke-related detrimental effects on the respiratory tract. However, despite the so-claimed reduced health risk, such products

Acknowledgements

This project was entirely supported by a grant from Antidoping Switzerland. The authors would also like to gratefully acknowledge Michaël Goy for his technical assistance.

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