Steroid profile and IRMS analysis of musk administration for doping control

Abstract

Musk, the dried secretion of the musk pod (sac) of adult male musk deer, has been used as traditional Chinese Medicine (TCM) in China and south-east Asian countries for thousands of years. Due to the anabolic steroid component in this TCM, musk preparations have been included in the list of medical products containing prohibited substances employed for doping by the State Food and Drug Administration of China. The application of musk pod formulation was claimed to be responsible for some adverse analytical findings (AAFs) in the 2011 FIFA Women's World Cup. Our preliminary study has suggested that musk ingestion did not lead to AAFs of doping control with the single dosage of 100 mg. However, the influences of musk administration in large and multi dosage are still unclear. The aim of this study is to further investigate the influences of musk administration for doping control. Wild and domestic deer musk samples were collected. The concentrations and δ 13 C-values of steroids in musk were analyzed. In an excretion study, 200 and 100 mg of wild and domestic deer musk samples were administrated by 29 subjects, respectively. Fluctuations in steroid profile could be observed, and the ratio of 5α-androstane-3α,17β-diol to 5β-androstane-3α,17β-diol was more sensitive than other parameters. In the IRMS test, the Δδ 13 C-value between endogenous reference compound and etiocholanolone was a sensitive parameter, and AAFs were obtained. It is the first time to confirm with excretion study that musk administration could lead to positive result of doping control.

Discrimination of wild and domestic deer musk using isotope ratio mass spectrometry

“…The steroids in domestic deer musk possess the δ 13 C value close to those of the steroids endogenously produced by human beings, which would make the IRMS test lose efficacy in doping control. The musk ingestion may influence the urinal steroid profiles and be controlled by athlete biological passport …”

“…For each musk sample, An, Etio, 5α‐diol, 5β‐diol, DHEA, EpiA, T, ET, 4‐AD, DHT, 5α‐3β,17α‐diol, 5α‐3β,17β‐diol, 5α‐dione, E, Ediol, Etriol, Preg, 17OH‐Preg, Prog, and Chol were analyzed . The specified chromatographic and mass spectrometric parameters for quantification are shown in Section S1 and Table S1 of Supplementary data.…”

“…The IRMS analysis was performed as previously reported . The carbon isotope ratio measurements were performed using a Thermo Scientific TRACE GC Ultra (Thermo Fisher Scientific, Bremen, Germany) coupled with a Delta‐V Advantage isotope ratio mass spectrometer (Thermo Fisher Scientific) via a combustion interface (GC Combustion III, Thermo Scientific).…”

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

“…Also, orally administered musk was reported to affect the urinary steroid profile as demonstrated in a comprehensive study conducted with 29 participants and musk specimens obtained from wild and domestic musk deer. Particularly the ratio of 5αAdiol/5βAdiol was influenced, and athletes should be made aware of the fact that products containing musk or musk extracts can result in AAFs in doping controls …”

The 35th Manfred Donike workshop on doping analysis

“…The entire breadth of analytes relevant to sports drug testing programmes, ranging from low molecular mass drugs to high molecular mass compounds, analyzed by mass spectrometric, biochemical, and immunological assays, was the subject of in‐depth discussions, with specific aspects receiving particular attention, as highlighted in this special issue. Among these core areas, new strategies employing isotope ratio mass spectrometry for steroidal as well as non‐steroidal target analytes were presented and new methods improving the management and/or the test spectrum for peptidic drugs such as erythropoietin (EPO) and the Transforming Growth Factor‐β (TGF‐β) inhibitor luspatercept were shown. Other contributions focused on drug and drug candidate metabolism studies employing diverse approaches including different in vitro and in vivo methodologies, which provided critical information on target analytes for future doping controls.…”