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Targeting the Administration of Ecdysterone in Doping Control Samples

View ORCID ProfileMaria Kristina Parr, Gabriella Ambrosio, Bernhard Wuest, Monica Mazzarino, Xavier de la Torre, Francesca Sibilia, Jan Felix Joseph, Patrick Diel, Francesco Botrè
doi: https://doi.org/10.1101/685230
Maria Kristina Parr
1Institute of Pharmacy, Freie Universität Berlin, Germany
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  • For correspondence: maria.parr@fu-berlin.de
Gabriella Ambrosio
1Institute of Pharmacy, Freie Universität Berlin, Germany
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Bernhard Wuest
2Agilent Technologies, Santa Clara CA, USA
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Monica Mazzarino
3Laboratorio Antidoping FMSI, Rome, Italy
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Xavier de la Torre
3Laboratorio Antidoping FMSI, Rome, Italy
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Francesca Sibilia
3Laboratorio Antidoping FMSI, Rome, Italy
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Jan Felix Joseph
1Institute of Pharmacy, Freie Universität Berlin, Germany
4Core Facility BioSupraMol, Department of Biology, Chemistry, Pharmacy, Freie Universitaet Berlin, Germany
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Patrick Diel
5Department for Molecular and Cellular Sports Medicine, Institute for Cardiovascular Research and Sports Medicine, German Sport University Cologne, Cologne, Germany
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Francesco Botrè
3Laboratorio Antidoping FMSI, Rome, Italy
6Department of Experimental Medicine, ‘Sapienza’ University of Rome, Italy
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Abstract

Purpose 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.

Methods Analytical properties of ecdysterone were evaluated using GC-QTOF-MS and LC-QTOF-MS. Its metabolism and elimination in human were studied using urines collected after administration.

Results 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 SPE 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.

Conclusion 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 two days after the administration of a single dose of ~50 mg.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted June 27, 2019.
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Targeting the Administration of Ecdysterone in Doping Control Samples
Maria Kristina Parr, Gabriella Ambrosio, Bernhard Wuest, Monica Mazzarino, Xavier de la Torre, Francesca Sibilia, Jan Felix Joseph, Patrick Diel, Francesco Botrè
bioRxiv 685230; doi: https://doi.org/10.1101/685230
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Targeting the Administration of Ecdysterone in Doping Control Samples
Maria Kristina Parr, Gabriella Ambrosio, Bernhard Wuest, Monica Mazzarino, Xavier de la Torre, Francesca Sibilia, Jan Felix Joseph, Patrick Diel, Francesco Botrè
bioRxiv 685230; doi: https://doi.org/10.1101/685230

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