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Home / Equine Research Bank / Drug Test Anal / Bioformation of boldenone and related precursors/metabolites...
Drug testing and analysis2019; 12(2); 215-229; doi: 10.1002/dta.2706

Bioformation of boldenone and related precursors/metabolites in equine feces and urine, with relevance to doping control.

Abstract: Boldenone (1-dehydrotestosterone) is an exogenous anabolic-androgenic steroid (AAS) but is also known to be endogenous in the entire male horse and potentially formed by microbes in voided urine, the gastrointestinal tract, or feed resulting in its detection in urine samples. In this study, equine fecal and urine samples were incubated in the presence of selected stable isotope labeled AAS precursors to investigate whether microbial activity could result in 1-dehydrogenation, in particular the formation of boldenone. Fecal matter was initially selected for investigation because of its high microbial activity, which could help to identify potential 1-dehydrogenated biomarkers that might also be present in low quantities in urine. Fecal incubations displayed Δ1-dehydrogenase activity, as evidenced by the use of isotope labeled precursors to show the formation of boldenone and boldione from testosterone and androstenedione, as well as the formation of Δ1-progesterone and boldione from progesterone. Unlabeled forms were also produced in unspiked fecal samples with Δ1-progesterone being identified for the first time. Subsequent incubation of urine samples with the labeled AAS precursors demonstrated that Δ1-dehydrogenase activity can also occur in this matrix. In all urine samples where labeled boldenone or boldione were detected, labeled Δ1-progesterone was also detected. Δ1-progesterone was not detected any non-incubated urine samples or following an administration of boldenone undecylenate to one mare/filly. Δ1-progesterone appears to be a candidate for further investigation as a suitable biomarker to help evaluate whether boldenone present in a urine sample may have arisen due to microbial activity rather than by its exogenous administration.
© 2019 John Wiley & Sons, Ltd.
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Publication Date: 2019-11-21 PubMed ID: 31654498DOI: 10.1002/dta.2706Google Scholar: Lookup
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Summary

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The research article investigates the role of microbes in transforming anabolic-androgenic steroid (AAS) precursors into boldenone, an endogenous AAS in male horses, in equine feces and urine. The findings suggest that microbial activity can lead to the formation of boldenone, and that Δ1-progesterone, a newly discovered metabolite, may serve as a biomarker indicating whether boldenone presence in urine resulted from microbes or external administration.

Understanding the Formation of Boldenone

  • The researchers dedicated their attention to boldenone, a steroid that not only originates from external administration but is also naturally present in the bodies of the male horses. They explored whether it could be generated by the activity of microbes in expelled urine, the intestinal system, or food, which could lead to its detection in urine samples.
  • The main experiment involved incubating horse fecal and urine samples in the presence of chosen stable isotope-labeled AAS precursors. The ultimate goal was to find out if microbial activity would lead to 1-dehydrogenation, a chemical process believed to result in the formation of boldenone.

Fecal Matter and the Formation of Boldenone

  • The researchers initially zeroed in on horse fecal matter, largely due to its high microbial activity. This made it an accurate platform to identify potential biomarkers that might occur in urine samples but in low quantities.
  • The results demonstrated that fecal matter displays Δ1-dehydrogenase activity, as proven by the use of isotope-labeled hormone precursors to show the creation of boldenone and boldione. They also found evidence of the generation of Δ1-progesterone and boldione from progesterone.
  • Interestingly, these forms were also produced in fecal samples that were not spiked with an isotope, indicating the naturally occurring processes in a horse’s body. Notably, Δ1-progesterone was recognized for the first time.

Investigating Δ1-dehydrogenase Activity in Urine

  • Looking at urine samples, the researchers concluded that Δ1-dehydrogenase activity could likewise take place in this matrix. Urine samples where labelled boldenone or boldione were detected also revealed the presence of labeled Δ1-progesterone.
  • Contrarily, Δ1-progesterone was not found in untreated urine samples or in samples taken after administration of boldenone undecylenate to a mare.

Implications and Further Research

  • This novel discovery of Δ1-progesterone emerging through microbial activity and its reoccurrence with boldenone in urine samples led to the conclusion that it could potentially serve as a biomarker. This could help in determining whether any boldenone detected in a urine sample was due to microbial activity or exogenous administration, providing useful insights for anti-doping control measures in equine sports.
  • Unfortunately, the current research doesn’t provide clear preventive or detection measures, indicating a need for subsequent studies to thoroughly explore the role of Δ1-progesterone as a boldenone biomarker.

Cite This Article

APA
Viljanto M, Kicman AT, Walker CJ, Wolff K, Muir T, Hincks P, Biddle S, Scarth J. (2019). Bioformation of boldenone and related precursors/metabolites in equine feces and urine, with relevance to doping control. Drug Test Anal, 12(2), 215-229. https://doi.org/10.1002/dta.2706

Publication

Drug testing and analysis
ISSN: 1942-7611
NlmUniqueID: 101483449
Country: England
Language: English
Volume: 12
Issue: 2
Pages: 215-229

Researcher Affiliations

Viljanto, Marjaana
  • LGC, Fordham, Cambridgeshire, UK.
  • Drug Control Centre, Analytical and Environmental Sciences Research Division, King's College London, UK.
Kicman, Andrew T
  • Drug Control Centre, Analytical and Environmental Sciences Research Division, King's College London, UK.
Walker, Christopher J
  • Drug Control Centre, Analytical and Environmental Sciences Research Division, King's College London, UK.
Wolff, Kim
  • Drug Control Centre, Analytical and Environmental Sciences Research Division, King's College London, UK.
Muir, Tessa
  • British Horseracing Authority, London, UK.
Hincks, Pamela
  • LGC, Fordham, Cambridgeshire, UK.
Biddle, Simon
  • LGC, Fordham, Cambridgeshire, UK.
Scarth, James
  • LGC, Fordham, Cambridgeshire, UK.

MeSH Terms

  • Anabolic Agents / analysis
  • Anabolic Agents / metabolism
  • Anabolic Agents / urine
  • Animals
  • Chromatography, Liquid
  • Doping in Sports
  • Feces / chemistry
  • Horses / physiology
  • Horses / urine
  • Male
  • Substance Abuse Detection
  • Tandem Mass Spectrometry
  • Testosterone / analogs & derivatives
  • Testosterone / analysis
  • Testosterone / metabolism
  • Testosterone / urine

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