Busted: How GC and MS Help Detect Prohibitive Substances in Sports

The beginning of the 2024 Major League Baseball (MLB) season, coupled with the National Collegiate Athletic Association (NCAA) basketball tournaments, makes the end of March and the beginning of April a great time for the average sports fan. But professional athletes often feel immense pressure to perform well, which can occasionally lead to cheating.

One of the biggest ways athletes cheat is by doping, which has been common since Ancient Greece. During the Olympics in Ancient Greece, athletes would use figs to try and improve their performance (1). With the advancement of pharmaceuticals and technology, doping has evolved. Athletes now inject performance-enhancing drugs (PEDs) or experiment with cocktails of drugs to try and gain a competitive edge over other athletes (1).

However, PEDs and other illegal drugs also come with deleterious side effects. Doping, in particular, could lead to hypertension, cardiovascular issues, and liver damage, to name a few (2). A review article published in Microchemical Journal explores how chromatography and mass spectrometry-based techniques have contributed to detecting doping in athletes and why they are the most reliable techniques for this analysis (2).

The authors of the study focused on more than 10 groups of PEDs in their study. With each group, they discussed the analytical techniques that have been used to detect these illicit drugs. For athletes, one of the main groups of PEDs that are most abused in athletic competitions are anabolic-androgenic steroids (AAS) and blood doping.

Mass spectrometry (MS) received much of the focus in the article, especially when it came to talking about analytical methods for detecting AAS. The authors discussed how liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) and gas chromatography-electron ionization–tandem mass spectrometry (GC-EI-MS/MS) are the two main chromatographic techniques used to detect AAS in athletes (3,4). What makes both these techniques effective for this purpose is their high sensitivity and specificity (2). LC-ESI-MS and GC-EI-MS are also effective at identifying endogenous steroids (2).

Other techniques have also been experimented with to detect AAS include GC–MS triple quadrupole mass spectrophotometric analysis and high-performance LC (HPLC) to monitor an urine long-term marker for consumption of AAS called dehydrochloromethyltestosterone (DHCMT). For example, a study looked at using the abovementioned techniques to identify metabolites of DHCMT (5). In another study, Wang and his team used HPLC–MS/MS and multiple reaction monitoring (MRM) to identify 48 metabolites, which shows that HPLC–MS/MS can be used to establish the metabolic profiles of athletes, helping governing bodies to determine whether athletes have taken PEDs (6).

GC and mass spectrometry-based techniques are going to continue to remain a part of the game, even though their contributions will often go unnoticed. MS has been effectively used to detect drug metabolites in routine doping controls (2). As research continues to advance for detecting illicit drugs, the harder it will be for athletes to gain an unfair competitive advantage. As a result, chromatographic techniques, from behind the scenes, are helping to maintain the integrity of sports competitions.

References

(1) Holt, R. I. G.; Erotokritou-Mulligan, I.; Sonksen, P. H. The History of Doping and Growth Hormone Abuse in Sport. Growth Hormone & IGF Res. 2009, 19 (4), 320–326. DOI: 10.1016/j.ghir.2009.04.009

(2) Wahi, A.; Nagpal, R.; Verma, S.; Narula, A.; et al. A Comprehensive Review on Current Analytical Approaches Used For the Control of Drug Abuse in Sports. Microchem. J. 2023, 191, 108834. DOI: 10.1016/j.microc.2023.108834

(3) Polet, M.; Van Gansbeke, W.; Albertsdottir, A. D. Gas chromatography-mass spectrometry analysis of non-hydrolyzed sulfated steroids by degradation product formation. Drug Test. Anal. 2019, 11, 1656–1665. DOI: 10.1002/dta.2606

(4) Zhang, Y.; Wu, X.; Wang, W.; Huo, J.; et al. Simultaneous Detection of 93 Anabolic Androgenic Steroids in Dietary Supplements Using Gas Chromatography Tandem Mass Spectrometry. J. Pharm Biomed. Anal. 2022, 211, 114619.

(5) Loke, S.; de la Torre, X.; Iannone, M. Controlled Administration of Dehydrochloromethyltestosterone in Humans: Urinary Excretion and Long-Term Detection of Metabolites for Anti-Doping Purpose. J. Steroid Biochem. Mol. Biol. 2021, 214, 105978. DOI: 10.1016/j.jsbmb.2021.105978

(6) Wang, Z.; Zhou, X.; Liu, X.; et al. A Novel HPLC-MRM Strategy to Discover Unknown and Long-Term Metabolites of Stanozolol for Expanding Analytical Possibilities in Doping Control. J. Chromatogr. B. 2017, 1040, 250–259. DOI: 10.1016/j.jchromb.2016.11.006