In October 2020 United Kingdom Anti-Doping (UKAD) has reported an anti-doping rule violation against the rugby player Carl Hone after his sample tested positive for the prohibited substance Boldenone in a low concentration. The Athlete filed a statement in his defence and he was heard for the National Anti-Doping Panel.

The Athlete gave a prompt admission and denied the intentional use of the substance. He explained that the only possible source of the Boldenone was horse waste with which he must have come into contact when doing carpentry work at a barn adjacent to some stabling.

The London Lab confirmed that the concentration found in the Athlete's system was very low and it could no exclude the possibility of the Athlete's theory being correct.

UKAD contended that the Athlete failed to establish that the violation was not intentional on the balance of probability based on the contamination with horse waste theory.

The Panel considers it very unlikely that the Athlete as a low level amateur player had injected himself with the substance for performance enhancing. Nevertheless the Panel concludes that the Athlete had committed an anti-doping rule violation and that he failed to provide sufficient evidence that on the balance of probability horse waste was in fact the source of the prohibited substance in his system.

Therefore the National Anti-Doping Panel decides on 6 May 2021 to impose a 4 year period of ineligibility on the Athlete starting on the date of the sample collection, i.e. on 6 February 2020.

EPO transgene detection in dried blood spots for antidoping application / Alexandre Marchand, Ingrid Roulland, Florian Semence, Magnus Ericsson

• Drug Testing and Analysis 13 (2021) 11-12 (November-December), p. 1888-1896
• Special Issue: The 39th Manfred Donike workshop on doping analysis
• PMID: 33961347
• DOI: 10.1002/dta.3059

Abstract

The modification of gene expression to treat diseases is a field of research with exponential growth. As doping in sport closely follows emerging therapies, a surveillance of the modification of gene expression to enhance performance is needed. The gene coding for erythropoietin (EPO) is one target of interest. Since 2010, several protocols have been proposed to identify EPO gene doping by focusing on the presence in blood of a transgene that differ in size from the endogenous gene sequence, normally found in the human DNA. In this work, our aim was to validate an easily applicable method for EPO gene doping detection in dried blood spots (DBS). We evaluated the detection of EPO transgene in 20-μl DBS after the spike of a plasmid carrying the EPO transgene in whole blood. Three different DBS were compared: Nucleic-Card™, Whatman® 903, and the volumetric 20-μl VAMS™. Detection was performed with real-time polymerase chain reaction (PCR) and validated with two Taqman assays (one commercial and one custom) specific for the EPO transgene. The initial testing procedure could be done using one assay (custom) and the confirmation using the second one (commercial Taqman) with a final check of the size of the PCR product. Starting from 20-μl dried blood, 1000 copies of EPO transgene could efficiently be detected with the three types of DBS, VAMS showing a slightly better sensitivity. No loss of sensitivity was observed after 1-month storage of DBS at room temperature. This method could be applied to DBS collected during doping controls and allows reanalysis.

Related cases:

• UKAD 2019 UKAD vs Mark Dry
  October 8, 2019
• UKAD 2019 UKAD vs Mark Dry - Appeal
  February 25, 2020
• UKAD 2021 Mark Dry vs UKAD - Appeal
  August 2, 2021
  
  

On 25 February 2020 the Appeal Panel of the National Anti-Doping Tribunal decided to impose a 4 year period of ineligibility on the Athlete Mark Dry for Tampering. The Athlete had provided a false account to United Kingdom Anti-Doping (UKAD) as to why he was not at the address at the time after a missed test in October 2018.

The Appeal Panel concluded that the deliberate provision of false information was committed by the Athlete with the intention of evading the operation of the Rules and it disagreed with the conclusion of the Tribunal in First Instance.

Hereafter in December 2020 the Athlete requested UKAD for reconsideration of his sanction in light of the 2021 World-Anti-Doping Code.

The Athlete invoked the principle of lex mitior and the provisions in the UK ADR Article 10.3.1(b) regarding  exceptional circumstances for the imposition of a reduced sanction. In addition the Athlete requested UKAD to start the sanction backdated to the date of the Doping Control, i.e on 15 October 2018.

UKAD considers that the Appeal Panel's comment, and the Athletics Integrity Unit's (AIU) agreement with that comment, constitute exceptional circumstances that trigger a discretion to reduce the Athlete's period of ineligibility to between 2 and 4 years, depending on his degree of Fault.

In view of the Athlete's conduct UKAD deems that the Athlete's Fault lies in the middel of the light degree of Fault, corresponding to a period of ineligibility of 28 months. Yet UKAD holds that there are no grounds to start the sanction backdated to the date of the Doping Control.

Therefore UKAD decides on 7 May 2021 to reduce the Athlete's period of ineligibility from 4 years to 28 months.

In November 2020 the Athletics Integrity Unit (AIU) of World Athletics has reported an anti-doping rule violation against the American Athlete Omar Craddock for his Whereabouts Filing Failures resulting in 2 Missed Tests within a 12 month period. After notification a provisional suspension was ordered. The Athlete filed a statement in his defence and he was heard for the World Athletics Disciplinary Tribunal.

The Athlete did not challenge his Filing Failure on 1 April 2020 or the Misses Test on 29 July 2020. The Athlete only contested the Missed Test on 20 August 2019 and retracted his previous admission in his email of 29 August 2019. He testified that he believed he updated his Whereabouts information on 19 August listing 20 August as a travel day. He asserted with evidence that due to to problems associated with the used software the 20 August 2019 travel day was incorrectly entered into the system.

World Athletics rejected the Athlete's explanation that the Athlete believed he had updated his Whereabouts information for 20 August 2019 as a travel day on 19 August 2019. He had simply forgotten to update his Whereabouts information for 20 August 2019. Furthermore there had been no login activity (or attempted login activity) to either the website or the mobile application by the Athlete between 13 August 2019 and 20 August 2019.

Considering the evidence in this case the Sole Arbitrator concludes that the Athlete is responsible for 3 Whereabouts Failures within the 12 month period beginning 20 August 2019 and accordingly that he committed an anti-doping rule violation.

The Sole arbitrator finds that the Athlete had acted negligently as he had forgotten to update his Whereabouts while prior in his email of 29 August the Athlete indeed had admitted that he had missed the test on 20 August 2019.

Finally the Sole Arbitrator assessed the circumstances and the Athlete's of Fault in connexion with his 3 offences and deems that the Athlete's sanction can be reduced from 24 to 20 months.

Therefore the World Athletics Disciplinary Tribunal decides on 7 May 2021 to impose a 20 month period of ineligibility on the Athlete starting on the date of the provisional suspension, i.e. on 13 November 2020.

Cannabinoids, Phenolics, Terpenes and Alkaloids of Cannabis / Mohamed M. Radwan, Suman Chandra, Shahbaz Gul, Mahmoud A. ElSohly

• Molecules 26 (2021) 9 (8 May), 2774, p. 1-29
• PMID: 34066753
• PMCID: PMC8125862
• DOI: 10.3390/molecules26092774

Abstract

Cannabis sativa is one of the oldest medicinal plants in the world. It was introduced into western medicine during the early 19th century. It contains a complex mixture of secondary metabolites, including cannabinoids and non-cannabinoid-type constituents. More than 500 compounds have been reported from C. sativa, of which 125 cannabinoids have been isolated and/or identified as cannabinoids. Cannabinoids are C21 terpeno-phenolic compounds specific to Cannabis. The non-cannabinoid constituents include: non-cannabinoid phenols, flavonoids, terpenes, alkaloids and others. This review discusses the chemistry of the cannabinoids and major non-cannabinoid constituents (terpenes, non-cannabinoid phenolics, and alkaloids) with special emphasis on their chemical structures, methods of isolation, and identification.

The Use of Dietary Supplements in Fitness Practitioners : A Cross-Sectional Observation Study / Massimiliano Mazzilli, Filippo Macaluso, Stefano Zambelli, Pietro Picerno, Enzo Iuliano. - (Environmental Research and Public Health 18 (2021) 9 (9 May); 5005)

• PMID: 34065071
• PMCID: PMC8125933
• DOI: 10.3390/ijerph18095005

Abstract

This study was aimed at evaluating the prevalence of use of dietary supplements (DSs) among gym users and gym instructors involved in body shaping-oriented fitness training. Furthermore, this study aimed to verify whether differences existed in the prevalence and in the types of DSs used in both gym users and gym instructors involved in body shaping-oriented fitness competitions vs. those not involved in fitness competitions. A survey was distributed to 316 participants, composed of 89 gym instructors and 227 gym users of both genders aged 27.3 ± 7.7. Among these participants, 52 were involved in competitions and 248 were not, while 16 participants did not specify either way. The results showed a high prevalence in the use of DSs in the population considered, with 85.4% of the participants declaring they used DSs, with high heterogeneity in the numbers and in the combinations used. No differences were found between gym instructors and gym users, or between participants involved and those not involved in competitions. The results indicate that DSs are widely used by persons involved in body shaping-oriented fitness training. The results also suggest that the majority of the participants decided individually which DSs to use.

Keywords: body shaping; dietary supplements; exercise; food supplements; gyms; physical fitness.

In December 2020 the Jamaica Anti-Doping Commission (JADCO) has reported an anti-doping rule violation against the bodybuilder Deidre Lewis after her A and B samples tested positive for the prohibited substance Zeranol in a low concentration.

After notification  a provisional suspension was ordered. The Athlete filed a statement in her defence and she was heard for the Jamaica Independent Anti-Doping Disciplinary Panel.

The Athlete accepted the test results and admitted the violation. She denied the intentional use of the substance wheras she could not explain how the Zeranol had entered her system.

With support of an expert witness the Athlete asserted that the substance has a long excretion time and that the low concentration found in her samples could be consistent with natural food ingestion and not the result of supplement contamination.

The Panel finds that the presence of a prohibited substance has been established in the Athlete's sample and accordingly that she committed an anti-doping rule violation. The Panel considers that t is more likely that the violation was not intentional instead of that the violation was committed  intentionally.

Therefore the Jamaica Independent Anti-Doping Disciplinary Panel decides on 11 May 2021 to impose a 2 year period of ineligibility on the Athlete, starting on the date of the provisional suspension.

Comprehensive Isotope Ratio Metabolomics : Gas chromatography Isotope Ratio Mass Spectrometry of urinary metabolites and exhaled breath / Laura Rodas Sánchez, Pablo Rodríguez González, J. Ignacio García Alonso. - (Analytica Chimica Acta 1170 (2021) 25 July; 338606)

• PMID: 34090584
• DOI: 10.1016/j.aca.2021.338606

Abstract

We have developed an analytical procedure to measure the carbon isotopic composition of multiple compounds even when there is a partial overlap in the chromatographic profiles and applied this procedure to measure the carbon isotopic composition of different metabolites in human urine and exhaled breath. Method development and validation was performed with CRM IAEA-600 caffeine after calibration of the reference CO2 gas using a mixture of certified undecane, pentadecane and eicosane δ(13C) standards. The alternative data treatment procedure included the correction of time-lag between Faraday cup amplifiers (44 ms at mass 45 and -160 ms at mass 46), the calculation and correction of chromatographic isotope effects on each peak (isotope shifts) and the calculation of the isotope ratio for each compound using the linear regression slope procedure with data only at the top of the chromatographic peak. In that way, partial chromatographic overlap between different metabolites can be tolerated (resolution equal or higher than 1). The reproducibility (SD) of the carbon isotope composition of 93 metabolites in human urine (n = 8) from one volunteer was typically better than 0.5 δ(13C) (range 0.1-2.0 δ(13C), median 0.4 δ(13C)). The method was applied to follow the carbon isotope composition of different metabolites in human urine and exhaled breath after the oral administration of 100 mg of universally labelled 13C-glucose to another human volunteer. It was demonstrated that isotopically labelled compounds could be detected in both samples even 2 h after administration. So, the developed methodology can be applied to multiple types of samples containing a large number of partially overlapping analytes including environmental applications, anti-doping control or metabolomics studies, including the use of enriched isotope tracers.

Related case:

TJD-AD 2021-021 Appeal Decision - Football
September 14, 2021

In October 2019 the Brazilian Doping Control Authority (ABCD) has reported an anti-doping rule violation against the football player after his A and B samples tested positive for the prohibited substance Enobosarm (Ostarine). After notification a provisional suspension was ordered and the Athlete filed a statement in his defence.

The Athlete denied the intentional use of the substance and stated that all the supplements he had used were provided by his football club. At first the analysis of these supplements in the Rio Lab did not reveal prohibited substances. Yet analysis of another batch of these supplements from a compounding pharmacy, provided to the Athlete after the Doping Control in August 2019, confirmed the presence of traces of Ostarine and Anastrozole in two products.

ABCD disputed the admissibility of the analysis of the second batch due to these supplements were compounded and provided to ABCD after the Doping Control and due to these supplements were delivered in unsealed condition.

The Rapporteur finds that the presence of a prohibited substance has been established in the Athlete's sample and accordingly that he committed an anti-doping rule violation.

Based on the evidence the Rapporteur concludes that the violation was not intentional and that a contaminated supplement was the source of the positive test.

The Rapporteur considers that the Notification after 2 months hampered the Athlete in finding the source of the contamination, whereas he was tested before without issues. Further the Athlete's conduct was considered and his degree of negligence since he failed to mention his supplements on the Doping Control Form

Therefore the TJD-AD decides on 14 May 2021 to impose a 2 year period of ineligibility on the Athlete, starting on the date of the sample collection, i.e. on 8 August 2019.

In a Case of Death Involving Steroids, Hair Testing is more Informative than Blood or Urine Testing / Pascal Kintz, Laurie Gheddar, Adeline Blanchot, Alice Ameline, Jean-Sébastien Raul. - (Journal of Analytical Toxicology (2021) 15 May)

• PMID: 33991187
• DOI: 10.1093/jat/bkab048

Abstract

A 59-year old male was found dead at home, with 2 empty vials of an oily preparation attributed to a manufacturer from East Europe. There was no label on the vial. The subject was a former weightlifter, also known as an anabolic steroids abuser. The local prosecutor ordered a body examination, which was unremarkable and allowed collecting femoral blood, urine and scalp hair (6 cm, brown). He was treated for cardiac insufficiency with quinidine. Biological specimens were submitted to standard toxicological analyses including a screening with LC-QToF, but also to a specific LC-MS/MS method for anabolic steroids testing. Ethanol was not found in both blood and urine. Quinidine blood concentration (791 ng/mL) was therapeutic. No drug of abuse was identified. In blood, testosterone was less that 1 ng/mL and no other steroid was identified. In urine, T/E was 1.56 and boldenone showed up at 9 ng/mL. The hair test results, performed on the whole length, demonstrated repetitive steroids abuse, including testosterone (140 pg/mg), testosterone propionate (605 pg/mg) and testosterone decanoate (249 pg/mg), but also boldenone (160 pg/mg), trenbolone (143 pg/mg) and metandienone (60 pg/mg). Given forensic laboratories have limited access to steroid urinary metabolites reference material due to specific regulations (to avoid testing athletes before anti-doping verifications), hair analyses seem to be the best approach to document anabolic agents abuse. Indeed, in hair, the target drug is the parent compound and, in addition, when compared to blood or urine, this matrix has a much larger window of detection. The pathologist concluded to cardiac insufficiency in a context involving repetitive abuse of anabolic drugs. This case indicates that more attention should be paid to anabolic steroids, in a context of sudden cardiac death.