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Bioformation of boldenone and related precursors/metabolites in equine feces and urine, with relevance to doping control.
Drug Testing and Analysis ( IF 2.6 ) Pub Date : 2019-11-21 , DOI: 10.1002/dta.2706
Marjaana Viljanto 1, 2 , Andrew T Kicman 2 , Christopher J Walker 2 , Kim Wolff 2 , Tessa Muir 3 , Pamela Hincks 1 , Simon Biddle 1 , James Scarth 1
Affiliation  

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.

中文翻译:

在马粪和尿液中马来烯酮和相关前体/代谢产物的生物形成,与兴奋剂控制有关。

Boldenone(1-dehydrotestosterone)是一种外源性合成代谢-雄激素类固醇(AAS),但在整个雄性马中也是内源性的,可能由排尿,胃肠道或饲料中的微生物形成,导致尿液中被检测到。在这项研究中,将马粪和尿液样品在选定的稳定同位素标记的AAS前体存在下进行温育,以研究微生物活性是否会导致1-脱氢,特别是形成马来烯酮。由于粪便具有很高的微生物活性,因此最初被选为研究对象,这可能有助于鉴定潜在的1-脱氢生物标志物,这些标志物也可能在尿液中少量存在。粪便培养显示出Δ1-脱氢酶活性,同位素标记的前体的使用证明了由睾丸激素和雄烯二酮形成的双烯酮和金刚烷,以及由孕酮形成的Δ1-孕酮和双胍。未加标的粪便样品中也产生未标记的形式,首次鉴定出Δ1-孕酮。随后将尿液样品与标记的AAS前体一起孵育,表明在该基质中也可以发生Δ1-脱氢酶活性。在所有检测到标记的双烯酮或金手指的尿液样本中,也检测到了标记的Δ1-孕酮。未在任何未孵育的尿液样品中或在一个母马/雌性动物中服用十一碳烯化十一碳烯酮后未检测到Δ1-孕酮。
更新日期:2019-11-21
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