Detection of stanozolol O- and N-sulfate metabolites and their evaluation as additional markers in doping control
Detection of stanozolol O- and N-sulfate metabolites and their evaluation as additional markers in doping control
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DOI：
10.1002/dta.2107
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发表时间：
2017-07-01
影响因子：
2.9
通讯作者：
Ventura, Rosa
中科院分区：
文献类型：
--
作者：
Balcells, Georgina;Matabosch, Xavier;Ventura, Rosa
斯坦诺唑尔（STAN）是运动药物检测中检出频率最高的合成代谢雄激素类固醇之一。STAN误用通常通过监测酶解后与葡萄糖醛酸偶联的代谢产物排出或通过液相色谱-串联质谱（LC-MS/MS）直接检测来检测。众所周知，一些先前描述的代谢物是在C17中形成硫酸盐偶联物的结果，这些偶联物在尿液中转化为它们的17-外显体。因此，磺化作用是STAN重要的II期代谢途径，目前尚未得到全面的研究。本研究的目的是通过LC-MS/MS对STAN代谢的硫酸盐组分进行评价，以确定潜在的长期代谢物，为兴奋剂控制提供价值。6名健康男性志愿者口服或肌肉注射STAN，并在给药后31天收集尿液样本。为了获得有助于制定分析策略（中性损失扫描、前体离子扫描和选定的反应监测获取模式）的质谱数据，以检测潜在的硫酸盐代谢物，对可作为标准品的I相代谢物进行磺化处理。采用LC-MS/MS对11种硫酸盐代谢物（M-I ~ M-XI）进行了检测和表征。本文为o -硫酸盐和n -硫酸盐的电离和破碎提供了有价值的数据。对于STAN，结果显示，与先前描述的长期代谢物（epistanozoll - n -glucuronide）相比，硫酸盐不能提高检测的回顾性。然而，硫酸盐代谢物可以作为STAN滥用检测的附加标记物。版权所有：John Wiley & Sons， Ltd。
Stanozolol (STAN) is one of the most frequently detected anabolic androgenic steroids in sports drug testing. STAN misuse is commonly detected by monitoring metabolites excreted conjugated with glucuronic acid after enzymatic hydrolysis or using direct detection by liquid chromatography-tandem mass spectrometry (LC-MS/MS). It is well known that some of the previously described metabolites are the result of the formation of sulfate conjugates in C17, which are converted to their 17-epimers in urine. Therefore, sulfation is an important phase II metabolic pathway of STAN that has not been comprehensively studied. The aim of this work was to evaluate the sulfate fraction of STAN metabolism by LC-MS/MS to establish potential long-term metabolites valuable for doping control purposes. STAN was administered to six healthy male volunteers involving oral or intramuscular administration and urine samples were collected up to 31 days after administration. Sulfation of the phase I metabolites commercially available as standards was performed in order to obtain MS data useful to develop analytical strategies (neutral loss scan, precursor ion scan and selected reaction monitoring acquisitions modes) to detect potential sulfate metabolites. Eleven sulfate metabolites (M-I to M-XI) were detected and characterized by LC-MS/MS. This paper provides valuable data on the ionization and fragmentation of O-sulfates and N-sulfates. For STAN, results showed that sulfates do not improve the retrospectivity of the detection compared to the previously described long-term metabolite (epistanozolol-N-glucuronide). However, sulfate metabolites could be additional markers for the detection of STAN misuse. Copyright (C) 2016 John Wiley & Sons, Ltd.