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Home / Equine Research Bank / Drug Test Anal / Detection of boldenone in the urine of female horses-ex vivo...
Drug testing and analysis2023; doi: 10.1002/dta.3521

Detection of boldenone in the urine of female horses-ex vivo formation versus administration.

Abstract: Boldenone is an anabolic-androgenic steroid (AAS) that is prohibited in equine sports. However, in certain situations, it is endogenous, potentially formed by the microbes in urine. An approach to the differentiation based on the detection of the biomarkers Δ1-progesterone, 20(S)-hydroxy-Δ1-progesterone and 20(S)-hydroxyprogesterone was assessed, and their concentrations were monitored in the urine of untreated female horses (n = 291) alongside boldenone, boldienone, testosterone and androstenedione. Using an ultra-sensitive analytical method, boldenone (256 ± 236 pg/mL, n = 290) and the biomarkers (Δ1-progesterone up to 57.6 pg/mL, n = 8; 20(S)-hydroxy-Δ1-progesterone 85.3 ± 181 pg/mL, n = 130; 20(S)-hydroxyprogesterone 43.5 ± 92.1 pg/mL, n = 158) were detected at low concentrations. The ex vivo production of Δ1-steroids was artificially induced following the storage of urine samples at room temperature for 7 days in order to assess the concentrations and ratios of the monitored steroids. The administration of inappropriately stored feed source also resulted in an increase in 20(S)-hydroxy-Δ1-progesterone concentrations and the biomarker ratios. Using the results from different datasets, an approach to differentiation was developed. In situations where the presence of boldenone exceeds a proposed action limit of 5 ng/mL, the presence of the biomarkers would be investigated. If Δ1-progesterone is above 50 pg/mL or if 20(S)-hydroxy-Δ1-progesterone is above 100 pg/mL with the ratio of 20(S)-hydroxy-Δ1-progesterone:20(S)-hydroxyprogesterone greater than 5:1, then this would indicate ex vivo transformation or consumption of altered feed rather than steroid administration. There remains a (small) possibility of a false negative result, but the model increases confidence that adverse analytical findings reported in female horses are caused by AAS administrations.
© 2023 John Wiley & Sons, Ltd.
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Publication Date: 2023-06-01 PubMed ID: 37264746DOI: 10.1002/dta.3521Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article examines if the detection of a prohibited anabolic-androgenic steroid (Boldenone) in female horse urine is linked to external administration or is an outcome of natural formation within the body. The study determines markers for differentiation and establishes a testing model.

Study Rationale and Overview

  • The research is premised on the controversy surrounding the source of Boldenone, an anabolic-androgenic steroid banned from use in horse racing. The study is driven by challenges where the steroid can be endogenously created as a natural byproduct of biological processes in the animal’s system, bringing doping accusations under scrutiny.

Research Methodology

  • The study monitored the concentrations of Boldenone, Boldienone, testosterone, androstenedione as well as three markers (Δ1-progesterone, 20(S)-hydroxy-Δ1-progesterone, and 20(S)-hydroxyprogesterone) in the urine of untreated female horses.
  • It utilised an ultra-sensitive analytical method to detect these substances in low concentrations in the samples. The process also tested the possibility of artificial manufacturing of Δ1-steroids from urine samples stored at everyday room temperatures for a week.

Outcomes and Observations

  • The study found an increase in biomarker concentrations when horse feed sources were not properly stored. The detection of elevated 20(S)-hydroxy-Δ1-progesterone concentrations and biomarker ratios suggested ex vivo transformation or consumption of altered feed, as opposed to intentional steroid administration.
  • The research crew developed an approach to differentiate between endogenous formation and external administration. It proposed a model that signaled for investigation into biomarker presence if Boldenone exceeded a certain action limit.

Conclusion and Implications

  • The study observed that detection of Δ1-progesterone above 50 pg/mL, or elevated levels of 20(S)-hydroxy-Δ1-progesterone (above 100 pg/mL) combined with a specific ratio of biomarkers indicated natural formation or altered feed consumption rather than steroid administration.
  • While a small possibility of a false negative was admitted, the overall confidence in adverse analytical findings indicating administration of anabolic-androgenic steroids in female horses increased significantly.

Cite This Article

APA
Viljanto M, Kaabia Z, Taylor P, Hincks P, Muir T, Habershon-Butcher J, Bailly-Chouriberry L, Scarth J. (2023). Detection of boldenone in the urine of female horses-ex vivo formation versus administration. Drug Test Anal. https://doi.org/10.1002/dta.3521

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English

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.
Hincks, Pamela
  • 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.

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