Detection and metabolic investigations of a novel designer steroid: 3‐chloro‐17α‐methyl‐5α‐androstan‐17β‐ol

The artificial increase of the physical capability of horses using drugs is well known in racing and other equine sports. Both illicit and therapeutic substances are regarded as prohibited substances in competition in most countries. Some countries make distinctions for a few, specific drugs which are, however, allowed for use in other countries. The primary objective in the case of doping control is the detection of any trace of drug exposure, either parent drug or any of its metabolites, using the most powerful analytical methods which are generally based on chromatographic/mass spectrometric techniques. Of major concern in horseracing is the absence of a single organization regulating the anti-doping framework; instead of this, individual racing authorities provide rules and regulations often resulting in variations in the applied doping control programmes of different countries. The aim of this paper is to review the recent literature (approximately from 2012 to mid-2016) to highlight the numerous and diverse challenges faced in doping control of racing and equestrian sports, including the detection of designer drugs (anabolic steroids or stimulants) and of other emerging prohibited substances, such as peptides and noble gases in horse urine and plasma. Moreover, the application of 'omics' techniques (especially of metabolomics) deserves attention for establishing possible fingerprints of drug abuse as well as the evolution of instrumental analysis resulting a powerful ally in the fight against doping in equine sports.

Section: Nickelmentioning

confidence: 99%

Challenges in detecting substances for equine anti‐doping

Fragkaki

Kioukia‐Fougia

Kiousi

et al. 2017

“…To model the liver, which is the principle organ involved in detoxification and metabolism of exogenous substances, preparations involving liver metabolic enzymes are amongst the most commonly used in vitro methods . Equine liver microsomes and S9 fraction are most commonly used to model equine metabolism due their ease of use and commercial availability.…”

Section: In Vitro Equine Steroid Metabolismmentioning

confidence: 99%

“…Typically, the reaction of glucose‐6‐phosphate with a glucose‐6‐phosphate dehydrogenase (G6PDH) enzyme is used, which regenerates NADH/NADPH as a by‐product of oxidation. This allows for the use of a catalytic amount of these co‐factors in the metabolism reaction . Recent reports have also demonstrated the practicality of using homogenized whole liver tissue to perform in vitro studies, in an effort to closely replicate in vivo metabolism .…”

Section: In Vitro Equine Steroid Metabolismmentioning

confidence: 99%

“…3α‐Chloro‐17α‐methyl‐5α‐androstan‐17β‐ol is an anabolic steroid containing C3‐chlorination, which was identified alongside the 3β‐chloro isomer (5:2 mixture) in red‐and‐black capsules containing white powder seized in 2012 by law‐enforcement in Queensland, Australia (Fig. ) . This compound had not been previously reported in the literature, and it appears to be the first instance of a C3‐halogenated anabolic steroid intended for doping purposes.…”

Section: α‐Chloro‐17α‐methyl‐5α‐androstan‐17β‐olmentioning

confidence: 99%

“…Owing to concerns about its chemical reactivity and potential toxicity, this compound was only studied using in vitro systems . Yeast, HEK293 and HuH7 androgen receptor bioassays have found the potency of the major 3α–chloro isomer to be similar to testosterone (87–147% potency), whilst the 3β‐isomer gave much lower potency (2‐9%).…”

Section: α‐Chloro‐17α‐methyl‐5α‐androstan‐17β‐olmentioning

confidence: 99%

See 2 more Smart Citations

A review of designer anabolic steroids in equine sports

Waller

McLeod

2016

Self Cite

In recent years, the potential for anabolic steroid abuse in equine sports has increased due to the growing availability of designer steroids. These compounds are readily accessible online in 'dietary' or 'nutritional' supplements and contain steroidal compounds which have never been tested or approved as veterinary agents. They typically have unusual structures or substitution and as a result may pass undetected through current anti-doping screening protocols, making them a significant concern for the integrity of the industry. Despite considerable focus in human sports, until recently there has been limited investigation into these compounds in equine systems. To effectively respond to the threat of designer steroids, a detailed understanding of their metabolism is needed to identify markers and metabolites arising from their misuse. A summary of the literature detailing the metabolism of these compounds in equine systems is presented with an aim to identify metabolites suitable for incorporation into screening protocols by anti-doping laboratories. The future of equine anti-doping research is likely to be guided by the incorporation of alternate testing matrices into routine screening, the improvement of in vitro technologies that can mimic in vivo equine metabolism, and the improvement of instrumentation or analytical methods that allow for the development of untargeted screening, and metabolomics approaches for use in anti-doping screening protocols.

Non‐targeted acquisition strategy for screening doping compounds based on GC‐EI‐hybrid quadrupole‐Orbitrap mass spectrometry: A focus on exogenous anabolic steroids

Cavalcanti

Rodrigues

Santos

et al. 2017

This is a first look at a non-targeted screening method based on Orbitrap gas chromatography-mass spectrometry (GC-MS) technology for a large number of banned compounds in sports found in urine, including exogenous anabolic steroids, β-agonists, narcotics, stimulants, hormone modulators, and diuretics. A simple sample preparation was processed in four steps: an enzymatic hydrolysis, liquid-liquid extraction, evaporation, and trimethylsilylation. All compounds were able to meet the World Anti-Doping Agency's sensitivity criteria with mass accuracies less than 1 ppm and with sufficient points across the peak by running the Orbitrap GC-MS in full-scan mode. In addition, we discuss our initial findings of using a full-scan selected ion monitoring-tandem mass spectrometry (SIM-MS/MS) approach as a way to obtain lower detection limits and reach desirable selectivity for some exogenous anabolic steroids.