Metabolism of boldione in humans by mass spectrometric techniques : detection of pseudoendogenous metabolites / Xavier de la Torre , Davide Curcio, Cristiana Colamonici, Francesco Molaioni, Francesco Botrè

• Drug Testing and Analysis 5 (2013) 11-12 (November-December), p. 834-842
• Special Issue: 31st Cologne workshop: Advances in sports drug testing
• PMID: 24259377
• DOI: 10.1002/dta.1567

Abstract

Boldione is an anabolic androgenic steroid (AAS) related to boldenone, androstenedione, and testosterone bearing two double bonds in C1 and C4 positions. Boldione is rapidly transformed to the well-known AAS boldenone, being both compounds included in the list of prohibited substances and methods published yearly by the World Anti-Doping Agency (WADA). After the administration of boldione to a male volunteer, the already described urinary metabolites of boldenone produced after reduction in C4, oxydoreduction in C3 and C17, and hydroxylation have been detected. In addition, minor new metabolites have been detected and their structure postulated after mass spectrometric analyses. Finally, the reduction of the double bound in C1 produces metabolites identical to the endogenously produced ones. A method based on gas chromatography coupled to isotope ratio mass spectrometry (GC/C/IRMS) after a urine sample purification by high performance liquid chromatography (HPLC) permitted to confirm the main synthetic like boldione/boldenone metabolite (17β-hydroxy-5β-androst-1-en-3-one) and boldenone at trace levels (< 5 ng/mL) and then to establish its synthetic or endogenous origin, and to determine the exogenous origin of metabolites with the same chemical structure of the endogenous ones. The detection of pseudoendogenous androgens of synthetic origin partially overlapped boldenone and its main metabolite detection, being an additional proof of synthetic steroids misuse. By the use of IRMS, the correct evaluation of the modifications of the steroid profile after the administration of synthetic AAS that could be converted into endogenous like ones is possible.

In December 2022 the Royal Dutch Billiards Federation (KNBB) reported an anti-doping rule violation against the Belgian billiard player after his sample tested positive for the prohibited substance Mesterolone.

Following notification a provisional suspension was ordered. The Person filed a statement in his defence and waived his right for a hearing. The case was settled by the ISR-KNBB Disciplinary Committee based on the written submissions of the parties.

The Person admitted the violation, accepted the test result en denied the intentional use of the substance. He recognized his fault and asserted that he had cooperated with the proceedings.

With evidence he demonstrated that the substance was used as medication to fulfil his desire to have children. Eight years ago for his first attempt to have childeren the Mesterone had also been used as prescribed medication.

Because prior he had been unware that the substance was prohibited the Person thereupon made an application for a retrospective TUE after the positive test. However this TUE application was rejected by NADO Flanders.

The Doping Authority Netherlands accepted that the Person's violation was not intentional and deemed that there are no grounds for a further reduced sanction.

The Doping Authority acknowledged that in August 2022 the KNBB and the Person not had been timely notified about the positive test. Because of this failure it proposed to start the Person's sanction on 6 August 2022.

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

The Committee determines that the Person's anti-doping rule violation was not intentional and that he had demonstrated how the substance had entered his system. However the Committee establishes that the Person had failed to check his medication, it was also used unprescribed and his retrospective TUE application was dismissed by NADO Flanders.

The Committee agrees that the imposed sanction can start backdated on 6 August 2022 because the KNBB could have ordered timely a provisional suspension on that date. However the KNBB, nor the Person, had been notified timely on that date by the Doping Autoritiy about the positive test.

Therefore the ISR-KNBB Disciplinary Committee decides on 5 September 2023 to impose a 2 year period of ineligibility on the Person, starting backdated on 6 August 2022, without disqualification of his results.

Fees and expenses for this committee shall be borne by the Person.

In January 2023 the International Tennis Integrity Agency (ITIA), on behalf of the International Tennis Federation (ITF), reported an anti-doping rule violation against the American tennis player Sydney Dorcil after her A and B samples tested positive for the prohibited substance Boldenone in a high concentration.

Following notification a provisional suspension was ordered. The Athlete filed a statement in her defence and she was heard for the ITF Independent Tribunal.

The Athlete admitted the violation and denied the intentional use of the substance. She asserted that she acted carefully with her supplements and medication she used.

The Athlete believed that the Boldenone entered her system through a contaminated B12 plus lipotropics injection that she received at a Lab in October 2022. She looked at all the ingredients before receiving the injection to assure herself that it did not contain any prohibited substances.

The Athlete ruled out meat contamination and contamination of her supplements and self-medication. The injection had to be the source.

The ITIA contended that the Athlete failed to establish with sufficient and corroborating evidence that a contaminated B12 injection was the source of the positive tests. It deemed that the filed evidence only shows that some veterinary medicines were compounded by the supplier in question, not that Boldenone was on the premises, nor that they had manufactured B12 injections.

In this case the Panel assessed and addressed the evidence and assertions of the Parties and their expert witnesses. Ultimately the Panel concludes that the Athlete failed to demonstrate that the violation was not intentional.

Further the Panel deems that the Athlete failed to meet the burden of establishing the source of the prohibited substance. The Panel holds that the Athlete was not a credible witness due to the inconsistencies between her evidence orally and in her statements.

Therefore the Panel decides on 6 July 2023 to impose a 4 year period of ineligibility on the Athlete, starting on the date of the provisional suspension, i.e. on 13 January 2023.

In November 2022 the International Tennis Integrity Agency (ITIA), on behalf of the International Tennis Federation (ITF), reported multiple anti-doping rule violations against the Polish tennis player Kamil Majchrzak.

In this case the presence had been established of the prohibited substances: LGD-4033 (Ligandrol), SARM S-22 (Ostarine) and GW0742 in the Athlete's A and B samples collected in:

• Sofia, 26 September 2022;
• Tokyo, 6 October 2022;
• Seoul, 12 October 2022; and
• Poland, 9 November 2022.

Following notification the Athlete admitted the violation, waived his right for a hearing, accepted a provisional suspension and the sanction proposed by the ITIA.

The Athlete denied that the violation was intentional and assumed that one of the prescribed supplement he had used could be the source of these banned substances. He asserted with evidence that he properly had researched the ingredients of his supplements before using.

Thereupon analysis of the supplement INTRA in a Laboratory revealed the presence of prohibited contaminants. Furthermore the Montreal Laboratory confirmed that the test results were consistent with the Athlete's use of this supplement at the material time.

In view of the evidence the ITIA accepts that the Athlete has demonstrated that the violation was not intentional as a result of his use of a contaminated supplement. Considering the Athlete's conduct in this case the ITIA determines that he acted with No Significant Fault or Negligence.

Therefore the ITIA decides on 16 June 2023 to impose a 13 month period of ineligibility on the Athlete, starting on the date of the provisional suspension, i.e. on 30 November 2022.

In November 2022 the International Tennis Integrity Agency (ITIA), on behalf of the International Tennis Federation (ITF), reported an anti-doping rule violation against the South African wheelchair tennis player Mariska Venter after her A and B samples tested positive for the prohibited substance Subutramine.

Following notification the Parathlete admitted the violation, waived her right for a hearing, accepted a provisional suspension and the sanction proposed by the ITIA.

The Parathlete denied the intentional use of the substance and assumed that her weight loss pills 4S Forever could be the source of the prohibited substance. She asserted with evidence that she properly had researched the ingredients of these pills before using.

Analysis of these pills in the Ghent Laboratory revealed the presence of Subutramine contaminants. Furthermore the Montreal Laboratory confirmed that the test results were consistent with the Parathlete's use of these pills at the material time.

In view of the evidence the ITIA accepts that the Parathlete has demonstrated that the violation was not intentional as a result of her use of contaminated weight loss pills. Considering the Parathlete's conduct in this case the ITIA determines that she acted with No Significant Fault or Negligence.

Therefore the ITIA decides on 15 March 2023 to impose a 12 month period of ineligibility on the Parathlete, starting on the date of the provisional suspension, i.e. on 14 December 2022.

In January 2022 the International Tennis Federation (ITF) reported an anti-doping rule violation against the Swedish tennis player Mikael Ymer for 3 Whereabouts Filing Failures within a 12 month period.

Yet, on 23 June 2022 the ITF Independent Tribunal determined that the Athlete had not committed an anti-doping rule violation in respect of his 3rd Whereabouts Failure.

Hereafter in July 2022 the ITF appealed the Decision of 23 June 2022 with the Court of Arbitration for Sport (CAS). The ITF requested the Panel to set aside the Appealed Decision and to impose a sanction of 2 years on the Athlete.

The ITF holds that its appeal centres on the Athlete's 3rd Whereabouts Failure and contended that the Doping Control Officer (DCO) on 7 November 2021 attempted to test the player during the 60-minute time slot by visiting the location specified for that timeslot. 

The ITF asserted that the DCO did what was reasonable in the circumstances to locate the Athlete. By contrast the Athlete denied his negligence and claimed that the DCO failed to do what was reasonable under the circumstances to locate the Athlete.

The Athlete stated that the filing task for his Whereabouts was delegated to a third party. He was not aware that on 7 November 2021 his stay in another tournament hotel in Roanne, France, did not correspond with his whereabouts filing for that day.

Following assessment of the evidence the Panel determines that de DCO did all that was required of him to locate the Athlete at the indicated hotel. Further the Panel deems that it is not the duty of the DCO to try to find the Athlete in another location than the Athlete's specified location.

The Panel is not satisfied on a balance of probability that the Athlete's behaviour was not negligent and did not cause or contribute to his failure to be available for testing.

The Panel finds it reasonable to expect that a tennis
player in the IRTP would not have delegated the filings task entirely to a third party, but  that such an Athlete would have verified the whereabouts filing made for that day, and
would thus have realized that his stay at the tournament hotel did not correspond with his whereabouts filing for that day.

Considering the Athlete's degree of fault the majority of the Panel finds that a sanction of 18 months must be imposed on the Athlete. Furthermore the Panel deems that there are no grounds to disqualify the Athlete's results prior to this Decision.

Therefore the Court of Arbitration for Sport decides on 17 July 2023 that:

1.) The Appeal filed by the International Tennis Federation against Mr Mikael Ymer with respect to the decision of the Independent Tribunal of 23 June 2023 is partially upheld.

2.) The decision rendered by the Independent Tribunal on 23 June 2023 is set aside.

3.) Mr Mikael Ymer is found to have committed an anti-doping violation under Article 2.4 of the Tennis Anti-Doping Programme.

4.) Mr Mikael Y mer is sanctioned with a period of ineligibility of 18 ( eighteen) months, starting from the date of notification of this Award.

5.) No results occurring between the time of the third missed test on 7 November 2021 and the date of this award are disqualified.

6.) The present arbitration proceedings shall be free of charge, except for the CAS Court Office fee of CHF 1,000 (one thousand Swiss Francs) paid by the International Tennis Federation, which is retained by the CAS.

7.) Each party shall bear its own costs and other expenses incurred in connection with these arbitration proceedings.

8.) All other applications and requests for relief are dismissed.

On 18 October 2022 the South African Institute for Drugfree Sport (SAIDS) reported an anti-doing rule violation against the South African boxer Zolani Tete after his A and B samples tested positive for the prohibited substance Stanozolol. The samples had been provided by the Athlete in Wembley in London on 2 July 2022 after a boxing match.

Following notification in October 2022 a provisional suspension was ordered while results management authority was referred to United Kingdom Anti-Doping (UKAD) in January 2023. The Athlete filed a statement in his defence and he was heard for the National Anti-Doping Panel (NADP).

The Athlete admitted the violation, accepted the test results and denied the intentional use of the substance. He assumed that contamination had caused the positive test results.

However the Athlete's attempts to find the source of the prohibited substance were unsucessful. Analysis of a supplement in question and his nail clippings in a laboratory revealed no prohibited substances.

In this case the Panel is troubled that there had been substantial delays attributed to the London Laboratory, SAIDS and UKAD. Because the Athlete was notified 3½ months after the sample collection this delay effected his opportunity to produce corroborating evidence in his defence.

Following assessment of the evidence the Panel concludes that the Athlete failed to demonstrate that the violation was not intentional, nor how the substance had entered his system. The Panel finds that there is lack of evidence in favour of the Athlete whereas the evidence is more consistent with a plausible motivation for using Stanozolol.

Considering the Athlete's conduct the Panel deems that he had acted with significant fault and negligence. Further the Panel already had determined that there had been substantial delays in this case not attributed to the Athlete.

Therefore the Panel decides on 9 August 2023 to impose a 4 year period of ineligibility on the Athlete, starting backdated on 30 July 2022.

In February 2023 the United Kingdom Anti-Doping (UKAD) reported an anti-doping rule violation against the rugby player Ryan Snowdeny after his sample tested positive for the prohibited substances Clenbuterol and Enobosarm (Ostarine).

Following notification the Athlete gave a prompt admission, waived his right for a hearing, accepted a provisional suspension and the sanction proposed by UKAD. The Athlete acknowleded that he had used a supplement containing Clenbuterol to lose weight.

UKAD deems that the Athlete's violation was intentional and determines that he shall receive a 1 year reduction to the period of ineligibility for his timely admission of the anti-doping rule violation.

Therefore UKAD decides on 7 July 2023 to impose a 3 year period of ineligibility on the Athlete, starting on the date of the provisional suspension, i.e. on 10 February 2023.

In February 2023 the United Kingdom Anti-Doping (UKAD) reported an anti-doping rule violation against the rugby player Rob Worrincy after his sample tested positive for the prohibited substances Enobosarm (ostarine), GW1516 and Ibutamoren.

Following notification the Athlete gave a prompt admission, waived his right for a hearing, accepted a provisional suspension and the sanction proposed by UKAD.

The Athlete acknowleded that he had used supplements to speed up recovery from an injury. He asserted that he was unaware that one of these supplements contained prohibited substances.

UKAD deems that the Athlete's violation was intentional and determines that he shall receive a 1 year reduction to the period of ineligibility for his timely admission of the anti-doping rule violation.

Therefore UKAD decides on 6 July 2023 to impose a 3 year period of ineligibility on the Athlete, starting on the date of the provisional suspension, i.e. on 24 February 2023.

Related case:

UKAD 2021 UKAD vs Dr Richard Freeman
July 20, 2023

Dr Richard Freeman is a former sports physician known for his work with British Cycling and Team Sky.

Following investigations United Kingdom Anti-Doping (UKAD) established that Dr Freeman in May 2011 had ordered 30 sachets of Testogel (Testosterone gel). These 30 sachets had been delivered to the National Cycling Centre in Manchester and were addressed to Dr Freeman.

Thereupon UKAD established that Dr Freeman had provided false statements about the purpose of the Testogel. Dr Freeman claimed that the Testogel was used as treatment for a 'non-rider' patient and that the Testogel had been returned to the supplier for destruction.

Consequently in December 2020 UKAD reported two anti-doping rule violations against Dr Freeman for possession of Testogel in 2011 and for Tampering in 2017 by providing false statements to UKAD in respect of ordering the Testogel.

After notification a provisional suspension was ordered. In April 2017 UKAD made a referral to the General Medical Council (GMC) in respect of UKAD's concerns relating to Dr Freeman's conduct and fitness to practise.

On 12 March 2021 the Medical Practitioners Tribunal Service (MPTS) determined that Dr Freeman had ordered Testogel and had lied to UKAD whilst under investigation. As a result the MPTS decided to erase Dr Freeman from the General Medical Council's Medical register.

Taking all these matters into account, the Tribunal considered that Dr Freeman’s behaviour is fundamentally incompatible with continued registration. The Tribunal has therefore determined that erasure is the only sufficient sanction which would protect patients, maintain public confidence in the profession and send a clear message to Dr Freeman, the profession and the public that his misconduct constituted behaviour unbefitting and incompatible with that of a registered doctor.

Hereafter the High Court dismissed on 16 January 2023 Dr Freeman's Appeal and confirmed the Appealed MPTS Decision. Thereupon UKAD could resume its investigations and the National Anti-Doping Panel (NADP) could conclude its proceedings against Dr Freeman.