Androgens, sports, and detection strategies for anabolic drug use

Walpurgis, Katja; Piper, Thomas; Thevis, Mario

doi:10.1016/j.beem.2021.101609

Cited by 8 publications

(6 citation statements)

References 116 publications

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“…The prominent role of anabolic agents, in particular AASs, in WADA's 2020 anti‐doping testing figures 101 has once more corroborated the importance of continuously improving test methods in sports drug testing laboratories. Comprehensive and extended analytical retrospectivity for AASs have been subject of continued research, but also advanced means facilitating the differentiation of natural and endogenously produced steroids from synthetic substances as well as enhancing the understanding of metabolic pathways depending on drug amounts or application routes 102 . In the light of the considerable health risks associated with AASs, both physically 38–43 and psychologically, 44–46 detection and containment in recreational as well as elite sport remains critical 103 .…”

Section: Anabolic Agentsmentioning

confidence: 99%

“…Comprehensive and extended analytical retrospectivity for AASs have been subject of continued research, but also advanced means facilitating the differentiation of natural and endogenously produced steroids from synthetic substances as well as enhancing the understanding of metabolic pathways depending on drug amounts or application routes. 102 In the light of the considerable health risks associated with AASs, both physically [38][39][40][41][42][43] and psychologically, [44][45][46] detection and containment in recreational as well as elite sport remains critical. 103 Here, the utility of luteinizing hormone (LH) was revisited and reported to represent a sensitive marker for current androgen abuse, 47 which, in concert with established targeted testing approaches, can contribute further insights into the prevalence and misuse of steroidal substances.…”

Section: Anabolic Agentsmentioning

confidence: 99%

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Annual banned‐substance review—Analytical approaches in human sports drug testing 2021/2022–

Thevis

Kuuranne

Geyer

2022

Drug Testing and Analysis

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Also in 2021/2022, considerable efforts were invested into advancing human sports drug testing programs, recognizing and taking into account existing as well as emerging challenges in anti-doping, especially with regard to substances and methods of doping specified in the World Anti-Doping Agency's 2022 Prohibited List. In this edition of the annual banned-substance review, literature on recent developments published between October 2021 and September 2022 is summarized and discussed.Focus is put particularly on enhanced analytical approaches and complementary testing options in human doping controls, appreciating the exigence and mission in anti-doping and, equally, the contemporary "new normal" considering, for example, the athlete's exposome versus analytical sensitivity and applicable anti-doping regulations for result interpretation and management.

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Section: Anabolic Agentsmentioning

confidence: 99%

Section: Anabolic Agentsmentioning

confidence: 99%

Annual banned‐substance review—Analytical approaches in human sports drug testing 2021/2022–

Thevis

Kuuranne

Geyer

2022

Drug Testing and Analysis

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“…Numerous synthetic AASs have been presented in the literature but only few of them have been fully evaluated and introduced in medical practice 3 . Due to their anabolic characteristics, AASs are the most frequently detected substances in sports drug testing for decades 4,5 . For monitoring purposes, World Anti‐Doping Agency (WADA) accredited doping control laboratories use complementary state‐of‐art gas chromatography‐(tandem) mass spectrometry (GC‐MS/(MS)) and/or liquid chromatography‐(tandem) mass spectrometry (LC‐MS/(MS)) instrumentation in order to target both parent AASs (when excreted in urine) and/or their metabolites after proper sample preparation 4 .…”

Section: Introductionmentioning

confidence: 99%

“…4,5 For monitoring purposes, World Anti-Doping Agency (WADA) accredited doping control laboratories use complementary state-of-art gas chromatography-(tandem) mass spectrometry (GC-MS/(MS)) and/or liquid chromatography-(tandem) mass spectrometry (LC-MS/(MS)) instrumentation in order to target both parent AASs (when excreted in urine) and/or their metabolites after proper sample preparation. 4 Knowledge of the metabolic fate for each one of the prohibited steroids is of paramount importance for the development of efficient detection methods, as AASs are extensively metabolized, notably in the liver, where conversion to their active form is sometimes required before they exert their biological action. 2 Because most AASs are extensively metabolized, monitoring of longterm AASs metabolites allows the confirmation of AASs misuse for a longer period of time.…”

mentioning

confidence: 99%

LC‐MS/(MS) confirmatory doping control analysis of intact phase II metabolites of methenolone and mesterolone after Girard's Reagent T derivatization

Angelis

Fragkaki

Kiousi

et al. 2023

Drug Testing and Analysis

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In the present study, the application and evaluation of Girard's Reagent T (GRT) derivatization for the simultaneous detection and significantly important identification of different phase II methenolone and mesterolone metabolites by LC‐MS/(MS) are presented. For the LC‐MS analysis of target analytes two complementary isolation methods were developed; a derivatization and shoot method in which native urine is diluted with derivatization reagent and is injected directly to LC‐MS and a liquid–liquid extraction method, using ethyl acetate at pH 4.5, for the effective isolation of both sulfate and glucuronide metabolites of the named steroids as well as of their free counterparts. For the evaluation of the proposed protocols, urine samples from methenolone and mesterolone excretion studies were analyzed against at least one sample from a different excretion study. Retention times, along with product ion ratios, were evaluated according to the WADA TD2021IDCR requirements, in order to determine maximum detection and identification time windows for each metabolite. Established identification windows obtained after LC‐MS/(MS) analysis were further compared with those obtained after GC‐MS/(MS) analysis of the same samples from the same excretion studies, for the most common analytes monitored by GC‐MS/(MS). Full validation was performed for the developed derivatization and shoot method for the identification of methenolone metabolite, 3α‐hydroxy‐1‐methylen‐5α‐androstan‐17‐one‐3‐glucuronide (mth3). Overall, the GRT derivatization presented herein offers a tool for the simultaneous sensitive detection of free, intact glucuronide and sulfate metabolites by LC‐MS/(MS) that enhance significantly the detection and identification time windows of specific methenolone and mesterolone metabolites for doping control analysis.

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“…To assess the plausibility of AAFs caused by intimate contact in general, information on the approximate concentration ranges of substances commonly abused in sports such as anabolic agents in sf would be helpful [13,14]. In this study, two different substances from the class of anabolic agents are investigated as to their appearance and concentrations in sf, with LGD-4033 representing a selective androgen receptor modulator (SARM) recently discussed in the context of an AAF related to intimate contact, and stanozolol (Stan) as a typical representative of anabolic steroids [5,15].…”

Section: Introductionmentioning

confidence: 99%

Investigations into the concentration and metabolite profiles of stanozolol and LGD-4033 in blood plasma and seminal fluid using liquid chromatography high-resolution mass spectrometry

et al. 2022

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Potential scenarios as to the origin of minute amounts of banned substances detected in doping control samples have been a much-discussed problem in anti-doping analysis in recent years. One such debated scenario has been the contamination of female athletes’ urine with ejaculate containing doping agents and/or their metabolites. The aim of this work was to obtain complementary information on whether relevant concentration ranges of doping substances are excreted into the ejaculate and which metabolites can be detected in the seminal fluid (sf) and corresponding blood plasma (bp) samples. A method was established to study the concentration and metabolite profiles of stanozolol and LGD-4033—substances listed under anabolic substances (S1) on the World Anti-Doping Agency’s Prohibited List—in bp and sf using liquid chromatography high-resolution mass spectrometry (LC-HRMS). For sf and bp, methods for detecting minute amounts of these substances were developed and tested for specificity, recovery, linearity, precision, and reliability. Subsequently, sf and bp samples from an animal administration study, where a boar orally received stanozolol at 0.33 mg/kg and LGD-4033 at 0.11 mg/kg, were measured. The developed assays proved appropriate for the detection of the target substances in both matrices with detection limits between 10 and 40 pg/mL for the unmetabolized drugs in sf and bp, allowing to estimate the concentration of stanozolol in bp (0.02–0.40 ng/mL) and in sf (0.01–0.25 ng/mL) as well as of LGD-4033 in bp (0.21–2.00 ng/mL) and in sf (0.03–0.68 ng/mL) post-administration. In addition, metabolites resulting from different metabolic pathways were identified in sf and bp, with sf resembling a composite of the metabolic profile of bp and urine. Graphical Abstract

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Androgens, sports, and detection strategies for anabolic drug use

Cited by 8 publications

References 116 publications

Annual banned‐substance review—Analytical approaches in human sports drug testing 2021/2022–

Annual banned‐substance review—Analytical approaches in human sports drug testing 2021/2022–

LC‐MS/(MS) confirmatory doping control analysis of intact phase II metabolites of methenolone and mesterolone after Girard's Reagent T derivatization

Investigations into the concentration and metabolite profiles of stanozolol and LGD-4033 in blood plasma and seminal fluid using liquid chromatography high-resolution mass spectrometry

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