Targeting the administration of ecdysterone in doping control samples
The phytosteroid ecdysterone was recently reported to enhance performance in sports and may thus be considered as a substance of relevance in anti-doping control. To trace back an administration of ecdysterone from urine samples, analytical properties have been investigated to assess its integration into initial testing procedures (ITP) in doping control laboratories.
Analytical properties of ecdysterone were evaluated using gas chromatography-quadrupole/time-of-flight-mass spectrometry (GC–QTOF–MS) and liquid chromatography (LC)–QTOF–MS. Its metabolism and elimination in human were studied using urines collected after administration.
The detectability of ecdysterone by GC–MS (after derivatization) and/or LC–MS(/MS) has been demonstrated and sample preparation methods were evaluated. Dilute-and-inject for LC–MS(/MS) or solid phase extraction using Oasis HLB for GC–MS or LC–MS were found most suitable, while liquid–liquid extraction was hampered by the high polarity of ecdysteroids.
Most abundantly, ecdysterone was detected in the post administration urines as parent compound besides the metabolite desoxy-ecdysterone. Additionally, desoxy-poststerone was tentatively assigned as minor metabolite, however, further investigations are needed.
An administration of ecdysterone can be targeted using existing procedures of anti-doping laboratories. Ecdysterone and desoxy-ecdysterone appeared as suitable candidates for integration in ITP. Using dilute-and-inject a detection of the parent compound was possible for more than 2 days after the administration of a single dose of ~ 50 mg.
KeywordsDoping control Monitoring program Ecdysterone Urine analysis LC–MS/MS GC–MS Accurate mass
The World Anti-Doping Agency is acknowledged for their financial support (research grant 18C18MP). Mrs. Maxi Wenzel, Freie Universitaet Berlin, is acknowledged for technical assistance.
Compliance with Ethical Standards
Conflict of interest
The authors declare no other conflict of interest.
All urine samples were anonymized and handled in accordance with the ethical standards of the Helsinki Declaration.
Informed consent form was signed by the volunteer.
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