Anabolic steroid use: patterns of use and detection of doping / Michael R. Graham, Bruce Davies, Fergal M. Grace, Andrew Kicman, Julien S. Baker

• Sports Medicine 38 (2008) 6 (June), p. 505-525
• PMID: 18489196
• DOI: 10.2165/00007256-200838060-00005

Abstract

Anabolic-androgenic steroids (AAS) were the first identified doping agents that have ergogenic effects and are being used to increase muscle mass and strength in adult males. Consequently, athletes are still using them to increase physical performance and bodybuilders are using them to improve size and cosmetic appearance. The prevalence of AAS use has risen dramatically over the last two decades and filtered into all aspects of society. Support for AAS users has increased, but not by the medical profession, who will not accept that AAS use dependency is a psychiatric condition. The adverse effects and potential dangers of AAS use have been well documented. AAS are used in sport by individuals who have acquired knowledge of the half-lives of specific drugs and the dosages and cycles required to avoid detection. Conversely, they are used by bodybuilders in extreme dosages with the intention of gaining muscle mass and size, with little or no regard for the consequences. Polypharmacy by self-prescription is prevalent in this sector. Most recently, AAS use has filtered through to 'recreational street drug' users and is the largest growth of drugs in this subdivision. They are taken to counteract the anorexic and cachectic effects of the illegal psychotropic street drugs. Screening procedures for AAS in World Anti-Doping Agency accredited laboratories are comprehensive and sensitive and are based mainly on gas chromatography-mass spectrometry, although liquid chromatography-mass spectrometry is becoming increasingly more valuable. The use of carbon isotope mass spectrometry is also of increasing importance in the detection of natural androgen administration, particularly to detect testosterone administration. There is a degree of contentiousness in the scenario of AAS drug use, both within and outside sport. AAS and associated doping agents are not illegal per se. Possession is not an offence, despite contravening sporting regulations and moral codes. Until AAS are classified in the same capacity as street drugs in the UK, where possession becomes a criminal offence, they will continue to attract those who want to win at any cost. The knowledge acquired by such work can only assist in the education of individuals who use such doping agents, with a view to minimizing health risks and hopefully once again create a level playing field in sport.

Effects of recombinant human LH and hCG on serum and urine LH and androgens in men / David J. Handelsman, Catrin Goebel, Amanda Idan, Mark Jimenez, Graham Trout, Rymantas Kazlauskas

• Clinical Endocrinology 71 (2009) 3 (September), p. 417-428
• PMID: 19170708
• DOI: 10.1111/j.1365-2265.2008.03516.x

Abstract

Context: The administration of gonadotrophins is prohibited in sport but the effect in men of recently available recombinant hCG and LH on serum and urine concentrations of gonadotrophins and androgens has not been systematically evaluated in the antidoping context.

Objective: To determine the time-course of recombinant LH (rhLH) and hCG (rhCG) on blood and urine hormone profiles in men to develop effective tests to detect rhLH and rhCG doping.

Design: Two randomized controlled studies with a 2 x 2 factorial design.

Setting: Academic research centre.

Participants: Healthy male volunteers aged 18-45 years.

Interventions: In the rhLH study, men were randomized into (i) either of two single doses of rhLH (75 IU or 225 IU), and (ii) suppression of endogenous LH and testosterone by nandrolone or no suppression. In the rhCG study, men were randomized into (i) either of two single doses of rhCG (250 or 750 microg), and (ii) suppression of endogenous LH and testosterone by nandrolone decanoate (ND) or no suppression. ND suppression comprised a single dose of 200 mg ND 3 days prior to, and in the rhCG study an additional dose 1 day after gonadotrophin injection.

Main outcome measures: Serum and urine hCG, LH, T, T : LH ratio, urine epitestosterone (E) and urine T : E ratio.

Results: Neither rhLH dose produced a significant increase in serum or urine LH or T or in the T : E or T : LH ratios regardless of ND-induced suppression of endogenous LH and T. Nor did an even higher dose (750 IU) in three healthy men with unsuppressed gonadal axis. These findings were confirmed with two different commercial LH immunoassays together with adjustment for any influence of urine sediment and dilution. Both rhCG doses produced a steep, dose-proportional increase in serum and urine hCG with increases in serum and urine T and suppression of serum and urine LH, regardless of hCG dose. Serum but not urine T was lowered by ND suppression. The T : LH ratio showed a progressive increase unrelated to rhCG dose or ND suppression, whereas both rhCG and ND suppression minimally increased T : E ratio.

Conclusions: Both rhCG doses produce a striking increase in serum hCG and T with suppression of serum LH but, at single doses up to 750 IU, rhLH has no influence on serum or urine LH or T. Effective rhLH doping, which relies on a sustained increases in endogenous T, would require much higher and more frequent daily rhLH doses. Use of LH immunoassays optimized for serum to detect rhLH doping by urine LH measurement requires more standardization and validation and, at present, is unreliable. The T : LH ratio is, however, a useful screening test for hCG doping although its utility requires further evaluation.

Position stand on androgen and human growth hormone use / Jay R. Hoffman, William J. Kraemer, Shalender Bhasin, Thomas Storer, Nicholas A. Ratamess, G. Gregory Haff, Darryn S. Willoughby, Alan D. Rogol

• Journal of Strength and Conditioning Research 23 (2009) 5 Suppl (August), p. S1-S59
• PMID: 19620932
• DOI: 10.1519/JSC.0b013e31819df2e6

Erratum in

• Journal of Strength and Conditioning Research 24(2010) 2 (February), p. 585

Abstract

Perceived yet often misunderstood demands of a sport, overt benefits of anabolic drugs, and the inability to be offered any effective alternatives has fueled anabolic drug abuse despite any consequences. Motivational interactions with many situational demands including the desire for improved body image, sport performance, physical function, and body size influence and fuel such negative decisions. Positive countermeasures to deter the abuse of anabolic drugs are complex and yet unclear. Furthermore, anabolic drugs work and the optimized training and nutritional programs needed to cut into the magnitude of improvement mediated by drug abuse require more work, dedication, and preparation on the part of both athletes and coaches alike. Few shortcuts are available to the athlete who desires to train naturally. Historically, the NSCA has placed an emphasis on education to help athletes, coaches, and strength and conditioning professionals become more knowledgeable, highly skilled, and technically trained in their approach to exercise program design and implementation. Optimizing nutritional strategies are a vital interface to help cope with exercise and sport demands (). In addition, research-based supplements will also have to be acknowledged as a strategic set of tools (e.g., protein supplements before and after resistance exercise workout) that can be used in conjunction with optimized nutrition to allow more effective adaptation and recovery from exercise. Resistance exercise is the most effective anabolic form of exercise, and over the past 20 years, the research base for resistance exercise has just started to develop to a significant volume of work to help in the decision-making process in program design (). The interface with nutritional strategies has been less studied, yet may yield even greater benefits to the individual athlete in their attempt to train naturally. Nevertheless, these are the 2 domains that require the most attention when trying to optimize the physical adaptations to exercise training without drug use.Recent surveys indicate that the prevalence of androgen use among adolescents has decreased over the past 10-15 years (). The decrease in androgen use among these students may be attributed to several factors related to education and viable alternatives (i.e., sport supplements) to substitute for illegal drug use. Although success has been achieved in using peer pressure to educate high school athletes on behaviors designed to reduce the intent to use androgens (), it has not had the far-reaching effect desired. It would appear that using the people who have the greatest influence on adolescents (coaches and teachers) be the primary focus of the educational program. It becomes imperative that coaches provide realistic training goals for their athletes and understand the difference between normal physiological adaptation to training or that is pharmaceutically enhanced. Only through a stringent coaching certification program will academic institutions be ensured that coaches that they hire will have the minimal knowledge to provide support to their athletes in helping them make the correct choices regarding sport supplements and performance-enhancing drugs.The NSCA rejects the use of androgens and hGH or any performance-enhancing drugs on the basis of ethics, the ideals of fair play in competition, and concerns for the athlete's health. The NSCA has based this position stand on a critical analysis of the scientific literature evaluating the effects of androgens and human growth hormone on human physiology and performance. The use of anabolic drugs to enhance athletic performance has become a major concern for professional sport organizations, sport governing bodies, and the federal government. It is the belief of the NSCA that through education and research we can mitigate the abuse of androgens and hGH by athletes. Due to the diversity of testosterone-related drugs and molecules, the term androgens is believed to be a more appropriate term for anabolic steroids.

Population based evaluation of a multi-parametric steroid profiling on administered endogenous steroids in single low dose / Pieter Van Renterghem, Peter Van Eenoo, Frans T. Delbeke

• Steroids 75 (2010) 13-14 (12 December), p. 1047-1057
• PMID: 20688095
• DOI: 10.1016/j.steroids.2010.06.013

Abstract

Steroid profiling provides valuable information to detect doping with endogenous steroids. Apart from the traditionally monitored steroids, minor metabolites can play an important role to increase the specificity and efficiency of current detection methods. The applicability of several minor steroid metabolites was tested on administration studies with low doses of oral testosterone (T), T gel, dihydrotestosterone (DHT) gel and oral dehydroepiandrosterone (DHEA). The collected data for all monitored parameters were evaluated with the respective population based reference ranges. Besides the traditional markers T/E, T and DHT, minor metabolites 4-OH-Adion and 6α-OH-Adion were found as most sensitive metabolites to detect oral T administration. The most sensitive metabolites for the detection of DHEA were identified as 16α-OH-DHEA and 7β-OH-DHEA but longest detection up to three days (after oral administration of 50 mg) was obtained with non-specific 5β-steroids and its ratios. Steroids applied as a gel had longer effects on the metabolism but were generally not detectable with universal decision criteria. It can be concluded that population based reference ranges show limited overall performance in detecting misuse of small doses of natural androgens. Although some minor metabolites provide additional information for the oral testosterone and DHEA formulations, the topical administered steroids could not be detected for all volunteers using universal reference limits. Application of other population based threshold limits did not lead to longer detection times.

Influence of multiple injections of human chorionic gonadotropin (hCG) on urine and serum endogenous steroids concentrations / Emmanuel Strahm, Pedro Marques-Vidal, François Pralong, Jiri Dvorak, Martial Saugy, Norbert Baume. - (Forensic Science International 213 (2011) 1-3 (10 December); p. 62-72)

• PMID: 21798680
• DOI: 10.1016/j.forsciint.2011.07.002

Abstract

Since it is established that human chorionic gonadotropin (hCG) affects testosterone production and release in the human body, the use of this hormone as a performance enhancing drug has been prohibited by the World Anti-Doping Agency. Nowadays, the only validated biomarker of a hCG doping is its direct quantification in urine. However, this specific parameter is subjected to large inter-individual variability and its determination is directly dependent on the reliability of hCG immunoassays used. In order to counteract these weaknesses, new biomarkers need to be evidenced. To address this issue, a pilot clinical study was performed on 10 volunteers submitted to 3 subsequent hCG injections. Blood and urine samples were collected during two weeks in order to follow the physiological effects on related compounds such as the steroid profile or hormones involved in the hypothalamo-pituitary axis. The hCG pharmacokinetic observed in all subjects was, as expected, prone to important inter-individual variations. Using ROC plots, level of testosterone and testosterone on luteinizing hormone ratio in both blood and urine were found to be the most relevant biomarker of a hCG abuse, regardless of inter-individual variations. In conclusion, this study showed the crucial importance of reliable quantification methods to assess low differences in hormonal patterns. In regard to these results and to anti-doping requirements and constraints, blood together with urine matrix should be included in the anti-doping testing program. Together with a longitudinal follow-up approach it could constitute a new strategy to detect a hCG abuse, applicable to further forms of steroid or other forbidden drug manipulation.

Strahm E, Marques-Vidal P, Pralong F, Dvorak J, Saugy M, Baume N. Influence of multiple injections of human chorionic gonadotropin (hCG) on urine and serum endogenous steroids concentrations. Forensic Sci Int. 2011 Dec 10;213(1-3):62-72. Epub 2011 Jul 27.

Current use of high-resolution mass spectrometry in drug screening relevant to clinical and forensic toxicology and doping control / Ilkka Ojanperä, Marjo Kolmonen, Anna Pelander

• Analytical & Bioanalytical Chemistry 403 (2012) 5 (May), p. 1203-1220
• PMID: 22302167
• DOI: 10.1007/s00216-012-5726-z

Abstract

Clinical and forensic toxicology and doping control deal with hundreds or thousands of drugs that may cause poisoning or are abused, are illicit, or are prohibited in sports. Rapid and reliable screening for all these compounds of different chemical and pharmaceutical nature, preferably in a single analytical method, is a substantial effort for analytical toxicologists. Combined chromatography-mass spectrometry techniques with standardised reference libraries have been most commonly used for the purpose. In the last ten years, the focus has shifted from gas chromatography-mass spectrometry to liquid chromatography-mass spectrometry, because of progress in instrument technology and partly because of the polarity and low volatility of many new relevant substances. High-resolution mass spectrometry (HRMS), which enables accurate mass measurement at high resolving power, has recently evolved to the stage that is rapidly causing a shift from unit-resolution, quadrupole-dominated instrumentation. The main HRMS techniques today are time-of-flight mass spectrometry and Orbitrap Fourier-transform mass spectrometry. Both techniques enable a range of different drug-screening strategies that essentially rely on measuring a compound's or a fragment's mass with sufficiently high accuracy that its elemental composition can be determined directly. Accurate mass and isotopic pattern acts as a filter for confirming the identity of a compound or even identification of an unknown. High mass resolution is essential for improving confidence in accurate mass results in the analysis of complex biological samples. This review discusses recent applications of HRMS in analytical toxicology.

Analytical aspects in doping control: challenges and perspectives / Flavia Badoud, Davy Guillarme, Julien Boccard, Elia Grata, Martial Saugy, Serge Rudaz, Jean-Luc Veuthey

• Forensic Science International 213 (2011) 1-3 (10 December), p. 49-61
• PMID: 21824736
• DOI: 10.1016/j.forsciint.2011.07.024

Abstract

Since the first anti-doping tests in the 1960s, the analytical aspects of the testing remain challenging. The evolution of the analytical process in doping control is discussed in this paper with a particular emphasis on separation techniques, such as gas chromatography and liquid chromatography. These approaches are improving in parallel with the requirements of increasing sensitivity and selectivity for detecting prohibited substances in biological samples from athletes. Moreover, fast analyses are mandatory to deal with the growing number of doping control samples and the short response time required during particular sport events. Recent developments in mass spectrometry and the expansion of accurate mass determination has improved anti-doping strategies with the possibility of using elemental composition and isotope patterns for structural identification. These techniques must be able to distinguish equivocally between negative and suspicious samples with no false-negative or false-positive results. Therefore, high degree of reliability must be reached for the identification of major metabolites corresponding to suspected analytes. Along with current trends in pharmaceutical industry the analysis of proteins and peptides remains an important issue in doping control. Sophisticated analytical tools are still mandatory to improve their distinction from endogenous analogs. Finally, indirect approaches will be discussed in the context of anti-doping, in which recent advances are aimed to examine the biological response of a doping agent in a holistic way.

Sports drug testing – an analyst's perspective / Graham J. Trout, Rymantas Kazlauskas

• Chemical Society Reviews 33 (2004) 1 (January), p. 1-13
• PMID: 14737504
• DOI: 10.1039/b201476a

Abstract

Sport plays a major role in the lives of many people, both for active participation and as entertainment. Sport is now a huge nationally and internationally based industry. The desire to win has led some athletes to resort to the use of performance enhancing drugs. With huge financial rewards now available in some sports the pressure to excel has grown. Some have argued that drug use should be given free rein, however most people are of the view that it is athletic prowess that should be applauded not the efficacy of various performance enhancing drugs. Apart from the obvious aspects of equality and fair play, the use of drugs is associated with significant health risks. In the 1960's the use of stimulants in sports such as cycling led to the death of at least one cyclist. Since 1968 the International Olympic Committee (IOC) has required all Olympic Games' host cities to provide laboratory facilities for the analysis and detection of performance enhancing drugs. There are now 29 IOC accredited laboratories throughout the world that routinely test samples from athletes for the presence of such drugs. The purpose of this tutorial review is to give an overview of drug testing procedures, including those that were used at the last summer Olympic Games in Sydney 2000, and the incorporation of the latest developments in analytical chemistry technology in the drug testing process. More recently, developments in biotechnology mean that the use of whole new classes of drugs are banned in sport, often requiring new methodologies and techniques for their analysis. The contest between those who wish to cheat and those who wish to maintain fair play in sport is an ongoing one.

Beyond sports-doping headlines: the science of laboratory tests for performance-enhancing drugs / Caroline K. Hatton 

• Pediatric Clinics of North America 54 (2007) 4 (August), p. 713-733
• PMID: 17723873
• DOI: 10.1016/j.pcl.2007.04.008

Abstract

This primer on urine analysis in sports-doping control is an overview with an emphasis on the main analytical chemistry technologies in use: gas chromatography-mass spectrometry (GC-MS), liquid chromatography-tandem mass spectrometry (LC-MS-MS), isotope ratio mass spectrometry detection of exogenous testosterone use, and isoelectric focusing detection of recombinant erythropoietin (EPO) use. Included are graphic examples of GC-MS selected ion monitoring and full scan, LC-MS-MS, and EPO test electropherogram data; a list of common acronyms; and answers to questions frequently asked about tampering and test accuracy.

Annual banned-substance review : analytical approaches in human sports drug testing / Mario Thevis, Tiia Kuuranne, Hans Geyer, Wilhelm Schänzer. - (Drug Testing and Analysis 4 (2012) 1 (January); p. 2-16)

• PMID: 22287289
• DOI: 10.1002/dta.415

Abstract

International anti-doping efforts are harmonized and regulated under the umbrella of the World Anti-Doping Code and the corresponding Prohibited List, issued annually by the World Anti-Doping Agency (WADA). The necessity for a frequent and timely update of the Prohibited List (as the result of a comprehensive consultation process and subsequent consensual agreement by expert panels regarding substances and methods of performance manipulation in sports) is due to the constantly growing market of emerging therapeutics and thus new options for cheating athletes to illicitly enhance performance. In addition, 'tailor-made' substances arguably designed to undermine sports drug testing procedures are considered and the potential of established drugs to represent a doping substance is revisited in light of recently generated information. The purpose of the annual banned substance review is to support doping controls by reporting emerging and advancing methods dedicated to the detection of known and recently outlawed substances. This review surveys new and/or enhanced procedures and techniques of doping analysis together with information relevant to doping controls that has been published in the literature between October 2010 and September 2011.