用GC-Orbitrap-HRMS研究了人肝脏S9模型中氧甲氧酮和美沙酮代谢物的表征和代谢谱。

IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Siying Zheng, Yuqi Ge, Xian Fang, Mengpan Liu, Haoyi Sun, Xiaojun Deng, Lei Liao
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引用次数: 0

摘要

体外代谢模型提供了一种手段来规避与人类给药研究相关的伦理问题,允许对人类代谢进行初步预测,同时产生高浓度的代谢物进行表征。作为世界反兴奋剂机构(WADA)禁用清单上的s1.1级合成代谢雄激素类固醇,氧甲氧酮和美雄酮近年来一直出现在不良分析结果(AAFs)中发现的前20种物质中,反映了它们在体育运动中的持续滥用模式。鉴于这些类固醇激素的外源性,其代谢产物属于兴奋剂控制的范围,因此代谢研究是反兴奋剂研究的一个重要方面。本研究以人肝脏S9为模型,采用气相色谱-轨道谱-高分辨率质谱(GC-Orbitrap-HRMS)技术对氧甲酮和美沙酮进行表征和代谢分析。利用GC-Orbitrap-HRMS全扫描模式检测游离代谢物和两种共轭代谢物,并将其与对照组进行比较,以确定体外培养过程中的代谢物。从EI质谱推断出可能的代谢物结构,并讨论了两种药物的代谢途径。体外鉴定了3种氧甲酮和5种美沙酮代谢物,其中2种代谢物OMT-M3 (2α,17α-甲基-5ξ-雄甾-3α,6β,17β-三醇)和MTS-M3 (2α,17α-二甲基-5ξ-雄甾-3α,12ξ,16ξ,17β-四醇)是根据近期人体体内代谢研究鉴定的新型代谢物。这些代谢物表现出不同的代谢途径,它们的结构通过体外和体内代谢分析得到证实。本研究全面评价了人肝脏S9模型在体外合成代谢类固醇代谢研究中的适用性,验证了新的人类代谢物,为该领域的未来研究提供了有价值的见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Characterization and metabolic profiling of oxymetholone and methasterone metabolites studied with human liver S9 model using GC-Orbitrap-HRMS for anti-doping purposes.

In vitro metabolic models provide a means to circumvent the ethical concerns associated with human administration research, allowing for preliminary predictions of human metabolism while generating high concentrations of metabolites for characterization. As S1.1-class anabolic androgenic steroids on the World Anti-Doping Agency (WADA) Prohibited List, oxymetholone and methasterone have consistently appeared in the top 20 substances identified in adverse analytical findings (AAFs) in recent years, reflecting their persistent abuse patterns in sports. Given their exogenous nature, the metabolites of these steroid hormones fall within the scope of doping control, making metabolic studies a crucial aspect of anti-doping research. In this study, human liver S9 fractions were employed as a model for the characterization and metabolic profiling of oxymetholone and methasterone via gas chromatography-orbitrap-high-resolution mass spectrometry (GC-Orbitrap-HRMS). The full scan mode of GC-Orbitrap-HRMS was utilized to detect free and two conjugated fractions of metabolites, comparing these with control groups to confirm the metabolites during in vitro incubation. Possible metabolite structures were inferred from EI mass spectra, and the metabolic pathways for both drugs were discussed. In vitro, three oxymetholone and five methasterone metabolites were identified, and among them, two metabolites, OMT-M3 (2α,17α-methyl-5ξ-androstan-3α,6β,17β-triol) and MTS-M3 (2α,17α-dimethyl-5ξ-androstane-3α,12ξ,16ξ,17β-tetrol), were characterized as novel metabolites based on recent human in vivo metabolic studies. These metabolites exhibited diverse metabolic pathways, and their structures were corroborated through complementary in vitro and in vivo metabolic analyses. This study provides a comprehensive evaluation of the applicability of the human liver S9 model in the metabolic studies of anabolic steroids in vitro, verifying novel human metabolites and providing valuable insights for future research in this field.

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来源期刊
CiteScore
8.00
自引率
4.70%
发文量
638
审稿时长
2.1 months
期刊介绍: Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.
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