Mass spectrometry in sports drug testing—Analytical approaches and the athletes' exposome

Abstract

Test methods in anti‐doping, most of which rely on the most modern mass spectrometric instrumentation, undergo continuous optimization in order to accommodate growing demands as to comprehensiveness, sensitivity, retrospectivity, cost‐effectiveness, turnaround times, etc. While developing and improving analytical approaches is vital for appropriate sports drug testing programs, the combination of today's excellent analytical potential and the inevitable exposure of humans to complex environmental factors, specifically chemicals and drugs at the lowest levels, has necessitated dedicated research, particularly into the elite athlete's exposome. Being subjected to routine doping controls, athletes frequently undergo blood and/or urine tests for a plethora of drugs, chemicals, corresponding metabolic products, and various biomarkers. Due to the applicable anti‐doping regulations, the presence of prohibited substances in an athlete's organism can constitute an anti‐doping rule violation with severe consequences for the individual's career (in contrast to the general population), and frequently the question of whether the analytical data can assist in differentiating scenarios of ‘doping’ from ‘contamination through inadvertent exposure’ is raised. Hence, investigations into the athlete's exposome and how to distinguish between deliberate drug use and potential exposure scenarios have become a central topic of anti‐doping research, aiming at supporting and consolidating the balance between essential analytical performance characteristics of doping control test methods and the mandate of protecting the clean athlete by exploiting new strategies in sampling and analyzing specimens for sports drug‐testing purposes.

The 41st Manfred Donike workshop on doping analysis

“…Conversely, the optimized sensitivity of analytical approaches regarding the growth hormone secretagogue capromorelin proved capable of detecting trace amounts of the drug in a doping control urine sample, which was attributable to a contamination scenario that an athlete was exposed to when administering the drug to a dog, 18 thus presenting another facet of the athlete's exposome. 19 Advancing the knowledge concerning peptide-based drug metabolites for sports drug testing purposes was conducted using in vitro and animal in vivo experiments, contributing new information for thymosin-β4, 20 the insulin-mimetic peptides S519 and S597, 21 and myostatin inhibitory peptides (e.g., DF-3 and DF-25). 22 Finally, and particularly relevant for result management and in avoiding unintentional anti-doping rule violations, higenamine excretion profiles in urine after ingestion of higenamine-containing fruit as well as enriched dietary supplement were investigated, suggesting that concomitantly observed markers of benzylisoquinoline alkaloid biosynthetic pathway in distinct abundance ratios are particularly valuable for a differentiation of diet-related exposure and doping scenarios.…”

“…While the technology proved principally useful, improvements in sensitivity are still required. Conversely, the optimized sensitivity of analytical approaches regarding the growth hormone secretagogue capromorelin proved capable of detecting trace amounts of the drug in a doping control urine sample, which was attributable to a contamination scenario that an athlete was exposed to when administering the drug to a dog, 18 thus presenting another facet of the athlete's exposome 19 …”

Annual banned‐substance review 16th edition—Analytical approaches in human sports drug testing 2022/2023

“…Metabolite profiles and patterns could play a major role also in differentiating doping scenarios from interindividual (cross-)contamination scenarios as argued in various cases of AAFs and the context of the athlete's exposome. 6 The occurrence of anabolic agents such as stanozolol and LGD-4033 (and corresponding metabolites) in The meat contamination issue concerning clenbuterol appears to persist as deduced from a total of 102 atypical findings (ATFs) regarding clenbuterol in 2021. 126 Supporting the analytical differentiation of "animal-processed" clenbuterol intake from doping scenarios has been desirable for many years, and considering the enantiomeric ratio of R-(À)-and S-(+)-clenbuterol in urine was discussed as a potential approach.…”

“…5 Determining the prevalence of doping in elite sport remains challenging, especially because analytical chemistry-based approaches have been discussed as underestimating the number of intentional ADRVs on the one hand while, at the same time, several AAFs have been attributed to exposure and contamination scenarios in the past. 6 Likewise, estimating doping prevalence by means of surveys proved complex as recently shown by the comparison of a data obtained via the Unrelated Question Model (UQM) or the Single Sample Count (SSC) strategy. 7 A considerable discrepancy was found when evaluating the surveys completed by athletes participating at the World Championships in Athletics 2011 and the 2011 Pan-Arab Games, where the prevalence of past year doping was reported with 43.6% and 57.1% (UQM), respectively, 21.2% and 10.6% (SSC).…”

“…Peptide hormones and their releasing factors, growth factors, and growth factor modulatorsRecent advances concerning analytes of the category of peptide hormones and corresponding releasing factors focused predominantly on gonadotrophin-releasing hormone (GnRH) and its synthetic analogs,140 human growth hormone (hGH), and growth hormone releasing hormones (GHRHs) and secretagogues (GHS). Investigations into the detection and the human in vivo metabolism of the GnRH analogs triptorelin and leuprorelin were conducted bySaardpun et al, who confirmed the formation and extended analytical retrospectivity of a previously in vitro-derived metabolite (referred to as triptorelin[5][6][7][8][9][10]) in humans 73. Urine samples from patients starting GnRH treatment with 11.25 mg of either leuprolide acetate or triptorelin pamoate were collected before and 3 and 6 h post-injection as well as after 30 days.…”