Detection and structural investigation of metabolites of stanozolol in human urine by liquid chromatography tandem mass spectrometry
Detection and structural investigation of metabolites of stanozolol in human urine by liquid chromatography tandem mass spectrometry
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DOI：
10.1016/j.steroids.2009.05.004
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发表时间：
2009-10-10
期刊：
影响因子：
2.7
通讯作者：
Delbeke, Frans T.
中科院分区：
文献类型：
--
作者：
Pozo, Oscar J.;Van Eenoo, Peter;Delbeke, Frans T.
研究了前驱体离子扫描LC-MS/MS法检测尿中司坦唑醇代谢物的适用性。对于未修饰A环或N环的司坦唑醇代谢物，选择了m/z为81的产物离子；对于N环和4-羟基-司坦唑醇代谢物，分别选择了m/z为97和145的产物离子。在此条件下，在阳性兴奋剂检测样本中发现了母药和多达15种代谢物。对服用司坦唑醇后收集的嵌合UPA-SCID小鼠样本的研究显示，存在另外4种代谢物。从产物离子光谱中获得的信息被用来建立一种检测19种化合物的SRM方法。将该SRM方法应用于几个掺杂正极样品。所有的代谢物在uPA-SCID小鼠样本和阳性的人类样本中都被检测到，而在被测试的空白样本中没有检测到；证实了所有被检测化合物的代谢性质。此外，将SRM方法应用于单个人体排泄研究，发现其中一种代谢物(4，16-二羟基-司坦唑醇)可以在负电离模式下检测到更长的时间，而不是通常用于兴奋剂分析中滥用司坦唑醇的筛查方法(3‘-羟基司坦唑醇、16β-羟基司坦唑醇和4-β-羟基司坦唑醇)。对几个阳性兴奋剂样本的应用证实了该代谢物在筛选司坦唑醇滥用方面的有效性。最后，根据UPLC-QTOF MS(C)2009 Elsevier Inc.用准确的质量测量的产物离子光谱，提出了每个被检测代谢物的初步结构。版权所有。
The applicability of LC-MS/MS in precursor ion scan mode for the detection of urinary stanozolol metabolites has been studied. The product ion at m/z 81 has been selected as specific for stanozolol metabolites without a modification in A- or N-rings and the product ions at m/z 97 and 145 for the metabolites hydroxylated in the N-ring and 4-hydroxy-stanozolol metabolites, respectively. Under these conditions, the parent drug and up to 15 metabolites were found in a positive doping test sample. The study of a sample from a chimeric uPA-SCID mouse collected after the administration of stanozolol revealed the presence of 4 additional metabolites. The information obtained from the product ion spectra was used to develop a SRM method for the detection of 19 compounds. This SRM method was applied to several doping positive samples. All the metabolites were detected in both the uPA-SCID mouse sample and positive human samples and were not detected in none of the blank samples tested; confirming the metabolic nature of all the detected compounds. In addition, the application of the SRM method to a single human excretion study revealed that one of the metabolites (4 xi,16 xi-dihydroxy-stanozolol) could be detected in negative ionization mode for a longer period than those commonly used in the screening for stanozolol misuse (3'-hydroxy-stanozolol, 16 beta-hydroxy-stanozolol and 4 beta-hydroxy-stanozolol) in doping analysis. The application of the developed approach to several positive doping samples confirmed the usefulness of this metabolite for the screening of stanozolol misuse. Finally, a tentative structure for each detected metabolite has been proposed based on the product ion spectra measured with accurate masses using UPLC-QTOF MS. (C) 2009 Elsevier Inc. All rights reserved.