Endogenous nature of estra-4,9-diene-3,17-dione in entire male horses.

IF 2.6 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Drug Testing and Analysis Pub Date : 2025-01-01 Epub Date: 2024-03-26 DOI:10.1002/dta.3685

Helen S M Ho, Emmie N M Ho, Wing-Tak Wong

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Abstract

Estra-4,9-diene-3,17-dione (dienedione) is an anabolic androgenic steroid (AAS) sold as a bodybuilding supplement. It is prohibited in both human and equine sports. With no report of 4,9-diene configuration in endogenous steroids, dienedione has long been considered a synthetic AAS. Nevertheless, the reoccurring detection of dienedione in colt (entire male horse) urine samples lead to the investigation of its possible endogenous nature in horses. This paper describes (i) the detection of naturally occurring dienedione in colts, (ii) the conjugation study of dienedione and (iii) the population study of free and glucuronide-conjugated dienedione in colt urine. Qualitative and quantitative analyses of dienedione content in colt urine were performed, employing liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Qualitative analyses showed that dienedione was endogenous in colt urine and mainly in the form of glucuronide conjugates. Glucuronidation of dienedione was believed to happen at 3-enol leading to dienedione-3-glucuronide. Upon the population study of free and glucuronide-conjugated dienedione in colt urine samples (n = 175), the mean ± SD was determined to be 2.5 ± 3.5 ng/ml. The population data fitted a normal distribution after a fifth root transformation with the exclusion of one outlier by Grubb's test. A possible in-house threshold was proposed at 30 ng/ml of free and glucuronide-conjugated dienedione in colt urine associated with a risk factor of 1 in 14,269 (with a degree of freedom of 173). This is the first report of endogenous dienedione in entire male horses and the approach for controlling its potential misuse by using a threshold is also presented.

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整匹雄马体内雌甾-4,9-二烯-3,17-二酮的内源性性质。

雌甾-4,9-二烯-3,17-二酮（二烯二酮）是一种同化雄性类固醇（AAS），作为健美补充剂出售。它在人类和马匹运动中均被禁用。由于没有关于内源性类固醇中存在 4,9- 二烯构型的报告，烯二酮一直被认为是一种合成的 AAS。然而，在马驹（整匹雄马）的尿样中再次检测到双烯二酮，这促使人们对其在马体内可能存在的内源性性质进行调查。本文介绍了(i)马驹体内天然存在的双烯二酮的检测，(ii)双烯二酮的共轭研究和(iii)马驹尿液中游离和葡萄糖醛酸共轭双烯二酮的群体研究。采用液相色谱-串联质谱法（UPLC-MS/MS）对马驹尿液中的双烯二酮含量进行了定性和定量分析。定性分析显示，马驹尿液中的双烯二酮是内源性的，主要以葡萄糖醛酸轭合物的形式存在。据认为，双烯二酮的葡萄糖醛酸化作用发生在 3-烯醇处，从而生成双烯二酮-3-葡萄糖醛酸苷。在对马驹尿样（n = 175）中游离和葡萄糖醛酸结合的双烯二酮进行群体研究后，确定其平均值（± SD）为 2.5 ± 3.5 纳克/毫升。总体数据经五次根变换后符合正态分布，经格拉布检验排除了一个离群值。提出了一个可能的内部阈值，即母马尿液中游离和葡萄糖醛酸结合的双烯二酮含量为 30 纳克/毫升，其风险系数为 14 269 分之 1（自由度为 173）。这是第一份关于整匹公马体内内源性双烯二酮的报告，报告还介绍了通过使用阈值来控制其潜在滥用的方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Drug Testing and Analysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

5.90

自引率

24.10%

发文量

191

审稿时长

2.3 months

期刊介绍： As the incidence of drugs escalates in 21st century living, their detection and analysis have become increasingly important. Sport, the workplace, crime investigation, homeland security, the pharmaceutical industry and the environment are just some of the high profile arenas in which analytical testing has provided an important investigative tool for uncovering the presence of extraneous substances. In addition to the usual publishing fare of primary research articles, case reports and letters, Drug Testing and Analysis offers a unique combination of; ‘How to’ material such as ‘Tutorials’ and ‘Reviews’, Speculative pieces (‘Commentaries’ and ‘Perspectives'', providing a broader scientific and social context to the aspects of analytical testing), ‘Annual banned substance reviews’ (delivering a critical evaluation of the methods used in the characterization of established and newly outlawed compounds). Rather than focus on the application of a single technique, Drug Testing and Analysis employs a unique multidisciplinary approach to the field of controversial compound determination. Papers discussing chromatography, mass spectrometry, immunological approaches, 1D/2D gel electrophoresis, to name just a few select methods, are welcomed where their application is related to any of the six key topics listed below.