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Drug testing and analysis2022; 14(5); 887-901; doi: 10.1002/dta.3240

Differentiation of boldenone administration from ex vivo transformation in the urine of castrated male horses.

Abstract: Boldenone is an anabolic-androgenic steroid that is prohibited in equine sports. However, in certain situations, it is endogenous or is believed to be formed by microbes in urine, and therefore, an approach for the differentiation is required. Following the identification of Δ1-progesterone and 20(S)-hydroxy-Δ1-progesterone as potential biomarkers of microbial activity, the presence of six steroids was investigated in the postrace urine of castrated male horses (geldings, n = 158). In line with endogenous findings from several other species when ultrasensitive methods are employed, boldenone was detected at low concentrations in all urine samples (27.0-1330 pg/ml). Furthermore, testosterone and androstenedione were detected in 157 samples (≤12,400 and 944 pg/ml, respectively), boldienone in two samples (≤22.0 pg/ml) and 20(S)-hydroxy-Δ1-progesterone in 20 samples (≤66.0 pg/ml). Δ1-Progesterone was not detected in any population samples analysed on arrival at the laboratory. The ex vivo transformation of boldienone, boldenone, androstenedione, Δ1-progesterone and 20(S)-hydroxy-Δ1-progesterone was induced following the storage of urine samples at room temperature for 7 days but not after refrigeration. Because the administration of inappropriately stored feed sources also resulted in an increase in 20(S)-hydroxy-Δ1-progesterone concentrations, a biomarker approach to distinguish steroid administrations was proposed. In situations where the presence of boldenone would exceed a proposed action limit, the presence of Δ1-progesterone and 20(S)-hydroxy-Δ1-progesterone would be investigated. If either Δ1-progesterone or 20(S)-hydroxy-Δ1-progesterone would exceed 50 and 100 pg/ml, respectively, for instance, then this would indicate ex vivo transformation or consumption of altered feed rather than steroid administration.
© 2022 John Wiley & Sons, Ltd.
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Publication Date: 2022-02-28 PubMed ID: 35178884DOI: 10.1002/dta.3240Google Scholar: Lookup
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Summary

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The research article investigates the detection of anabolic-androgenic steroid, Boldenone, in the urine of castrated male horses and the differentiation of its endogenous presence or ex vivo transformation. The presence of certain steroid compounds in urine after being induced by storage conditions or altered feed is also examined, aiming to provide a constructive approach to differentiate inappropriate steroid administration from natural occurrences.

Research Methodology and Findings

  • The research involved testing the urine of 158 geldings (castrated male horses) after participating in races to investigate the presence of six different steroid compounds, including Boldenone.
  • All urine samples were found to contain Boldenone, although at low concentrations. Other steroids such as Testosterone, Androstenedione, Boldienone, and 20(S)-hydroxy-Δ1-progesterone were identified in varying sample quantities.
  • One steroid, Δ1-Progesterone, was not detected in any samples analyzed immediately upon arrival at the laboratory.
  • The research also induced the ex vivo transformation of the aforementioned steroids by storing urine samples at room temperature for a week. This procedure did not generate any transformations when samples were kept refrigerated.

The Biomarker Approach for Steroid Detection

  • Observing that the concentration of 20(S)-hydroxy-Δ1-progesterone could either signify consumption of altered feeds or represents the steroid’s natural, in-body transformation, the research proposed a biomarker approach to distinguish between the two scenarios.
  • Under this approach, in situations where Boldenone concentration exceeds a predefined limit, the presence of Δ1-Progesterone and 20(S)-hydroxy-Δ1-progesterone would then be investigated.
  • If either Δ1-Progesterone or 20(S)-hydroxy-Δ1-progesterone concentrations exceeded 50 pg/ml and 100 pg/ml respectively, it would indicate that the steroids may have formed due to ex vivo transformation or consumption of altered feed sources, rather than from administration of the steroid.

Cite This Article

APA
Viljanto M, Kaabia Z, Taylor P, Muir T, Habershon-Butcher J, Bailly-Chouriberry L, Scarth J. (2022). Differentiation of boldenone administration from ex vivo transformation in the urine of castrated male horses. Drug Test Anal, 14(5), 887-901. https://doi.org/10.1002/dta.3240

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 14
Issue: 5
Pages: 887-901

Researcher Affiliations

Viljanto, Marjaana
  • LGC, Fordham, Cambridgeshire, UK.
Kaabia, Zied
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Taylor, Polly
  • LGC, Fordham, Cambridgeshire, UK.
Muir, Tessa
  • British Horseracing Authority, London, UK.
Habershon-Butcher, Jocelyn
  • British Horseracing Authority, London, UK.
Bailly-Chouriberry, Ludovic
  • GIE LCH, Laboratoire des Courses Hippiques, Verrières-le-Buisson, France.
Scarth, James
  • LGC, Fordham, Cambridgeshire, UK.

MeSH Terms

  • Anabolic Agents / urine
  • Androgens
  • Androstenedione
  • Animals
  • Doping in Sports
  • Horses
  • Male
  • Progesterone
  • Steroids
  • Testosterone / analogs & derivatives
  • Testosterone / urine

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Citations

This article has been cited 2 times.
  1. Cloteau C, Dervilly G, Loup B, Delcourt V, Kaabia Z, Bagilet F, Groseille G, Dauriac K, Fisher S, Popot MA, Garcia P, Le Bizec B, Bailly-Chouriberry L. Performance assessment of an equine metabolomics model for screening a range of anabolic agents. Metabolomics 2023 Apr 7;19(4):38.
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  2. Skoupá K, Šťastný K, Sládek Z. Anabolic Steroids in Fattening Food-Producing Animals-A Review. Animals (Basel) 2022 Aug 18;12(16).
    doi: 10.3390/ani12162115pubmed: 36009705google scholar: lookup
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