A novel HPLC-MRM strategy to discover unknown and long-term metabolites of stanozolol for expanding analytical possibilities in doping-control

In doping control analysis, the characterization of urinary steroid metabolites is of high interest for a targeted and long-term detection of prohibited anabolic androgenic steroids (AAS). In this work, the structure of a long-term metabolite of dehydrochloromethyltestosterone (DHCMT) was elucidated. Altogether, 8 possible metabolites with a 17α-methyl-17β-hydroxymethyl - structures were synthesized and compared to a major DHCMT long-term metabolite detected in reference urine excretion samples. The confirmed structure of the metabolite was 4α-chloro-18-nor-17β-hydroxymethyl-17α-methyl-5α-androst-13-en-3α-ol.

Section: Introductionmentioning

confidence: 99%

Unambiguous identification and characterization of a long‐term human metabolite of dehydrochloromethyltestosterone

Forsdahl

Geisendorfer

Göschl

et al. 2018

“…In the tanks where stanozolol was administered, it is possible to observe several peaks that indicate the production of the same metabolites of stanozolol as observed in human urine. All m/z values described for the stanozolol metabolites (Table ) were found in the tanks treated with stanozolol, including m/z 327.24309 and 361.24856, which have been indicated as targets for anti‐doping control because of their longer urinary excretion . Like sibutramine, it was already possible to observe the production of stanozolol metabolites in the treated tanks after 24 hours of administration (data not shown), and no peaks were observed in the negative controls.…”

Section: Resultsmentioning

confidence: 87%

“…AAS is a class of synthetic substances normally derived from testosterone, which is an endogenous androgen and protein anabolic hormone . Because of this, AAS is an important class of performance‐enhancing drugs with the potential for misuse in sports .…”

Section: Introductionmentioning

confidence: 99%

“…Because of this, AAS is an important class of performance‐enhancing drugs with the potential for misuse in sports . Stanozolol is an exogenous anabolic steroid designed from testosterone, and like sibutramine, it is prohibited by WADA . Stanozolol is intensively metabolized and its urinary metabolites can be detected much longer than the parent compound.…”

Section: Introductionmentioning

confidence: 99%

“…Stanozolol is intensively metabolized and its urinary metabolites can be detected much longer than the parent compound. The main phase‐I metabolic products that are important for human doping control purposes are 3′‐hydroxy‐stanozolol, 4β‐hydroxy‐stanozolol, 16β‐hydroxy‐stanozolol and 4,16‐dihydroxy‐stanozolol, which are excreted in human urine as glucuronide conjugates …”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Is zebrafish (Danio rerio) a tool for human‐like metabolism study?

Anselmo

Sardela

Matias

et al. 2017

One of the greatest challenges in anti-doping science is the large number of substances available and the difficulty in finding the best analytical targets to detect their misuse. Therefore, metabolism studies involving prohibited substances are fundamental. However, metabolism studies in humans could face an important ethical bottleneck, especially for non-approved substances. An emerging model for metabolism assessment is the zebrafish, due to its genetic similarities with humans. In the present study, the ability of adult zebrafish to produce metabolites of sibutramine and stanozolol, substances with a well-known metabolism that are widely used as doping agents in sports, was evaluated. They represent 2 of the most abused classes of doping agents, namely, stimulants and anabolic steroids. These are classes that have been receiving attention because of the upsurge of synthetic analogues, for which the side effects in humans have not been assessed. The samples collected from the zebrafish tank water were hydrolysed, extracted by solid-phase extraction, and analysed by liquid chromatography with high resolution mass spectrometry (LC-HRMS). Adult zebrafish could produce several sibutramine and stanozolol metabolites, including demethylated, hydroxylated, dehydroxylated, and reduced derivatives, all of which have already been detected in human urine. This study demonstrates that adult zebrafish can absorb, oxidise, and excrete several metabolites in a manner similar to humans. Therefore, adult zebrafish seem to be a very promising tool to study human-like metabolism when aiming to find analytical targets for doping control.

Annual banned‐substance review: Analytical approaches in human sports drug testing

Thevis

Kuuranne

Geyer

2017

Several high-profile revelations concerning anti-doping rule violations over the past 12 months have outlined the importance of tackling prevailing challenges and reducing the limitations of the current anti-doping system. At this time, the necessity to enhance, expand, and improve analytical test methods in response to the substances outlined in the World Anti-Doping Agency's (WADA) Prohibited List represents an increasingly crucial task for modern sports drug-testing programs. The ability to improve analytical testing methods often relies on the expedient application of novel information regarding superior target analytes for sports drug-testing assays, drug elimination profiles, alternative test matrices, together with recent advances in instrumental developments. This annual banned-substance review evaluates literature published between October 2016 and September 2017 offering an in-depth evaluation of developments in these arenas and their potential application to substances reported in WADA's 2017 Prohibited List.