Detection of reactive metabolites using isotope-labeled glutathione trapping and simultaneous neutral loss and precursor ion scanning with ultra-high-pressure liquid chromatography triple quadruple mass spectrometry.
Detection of reactive metabolites using isotope-labeled glutathione trapping and simultaneous neutral loss and precursor ion scanning with ultra-high-pressure liquid chromatography triple quadruple mass spectrometry.
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DOI：
10.1021/ac504737x
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发表时间：
2015-04-07
影响因子：
7.4
通讯作者：
van Breemen, Richard B.
中科院分区：
文献类型：
--
作者：
Huang, Ke;Huang, Lingyi;van Breemen, Richard B.
在美国，超过60%的黑盒警告和药物从市场上撤回是由于药物代谢活化为亲电物质。使用谷胱甘肽（GSH）捕获反应性代谢物，并使用高效液相色谱-串联质谱法（HPLC-MS/MS）或高效液相色谱-高分辨率质谱法（质量缺陷过滤）进行分析，使代谢活化筛选成为药物开发期间的常规操作。然而，目前基于MS的方法不能检测复杂混合物中存在的所有GSH缀合物，特别是植物膳食补充剂提取物中存在的那些。为了克服这些限制，开发了一种基于快速三重四极杆质谱仪的方法，该方法可以在单次分析中检测带正电荷和带负电荷的GSH缀合物，而不需要预先了解潜在缀合物的元素组成，同时避免假阳性。该方法利用UHPLC代替HPLC以缩短分离时间并提高灵敏度，掺入稳定同位素标记的GSH以避免假阳性，并使用快速极性切换电喷雾MS/MS来检测形成正离子和/或负离子的GSH缀合物。一般的新方法，然后被用来测试甘草膳食补充剂光甘草，发现形成多个GSH结合物后代谢活化。在甘草测定中发现的GSH缀合物中有异甘草素和光甘草定的缀合物，光甘草定是细胞色素P450酶的不可逆抑制剂。
Metabolic activation of drugs to electrophilic species is responsible for over 60% of black box warnings and drug withdrawals from the market place in United States. Reactive metabolite trapping using glutathione (GSH) and analysis using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) or HPLC with high resolution mass spectrometry (mass defect filtering) have enabled screening for metabolic activation to become routine during drug development. However, current MS-based approaches cannot detect all GSH conjugates present in complex mixtures, especially those present in extracts of botanical dietary supplements. To overcome these limitations, a fast triple quadrupole mass spectrometer-based approach was developed that can detect positively and negatively charged GSH conjugates in a single analysis without the need for advance knowledge of the elemental compositions of potential conjugates and while avoiding false positives. This approach utilized UHPLC instead of HPLC to shorten separation time and enhance sensitivity, incorporated stable-isotope labeled GSH to avoid false positives, and used fast polarity switching electrospray MS/MS to detect GSH conjugates that form positive and/or negative ions. The general new method was then used to test the licorice dietary supplement Glycyrrhiza glabra which was found to form multiple GSH conjugates upon metabolic activation. Among the GSH conjugates found in the licorice assay were conjugates with isoliquiritigenin and glabridin, which is an irreversible inhibitor of cytochrome P450 enzymes.
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影响因子：
7.4
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