Detection and structural characterization of glutathione-trapped reactive metabolites using liquid chromatography-high-resolution mass Spectrometry and mass defect filtering
Detection and structural characterization of glutathione-trapped reactive metabolites using liquid chromatography-high-resolution mass Spectrometry and mass defect filtering
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DOI：
10.1021/ac071119u
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发表时间：
2007-11-01
影响因子：
7.4
通讯作者：
Humphreys, W. Griffith
中科院分区：
文献类型：
--
作者：
Zhu, Mingshe;Ma, Li;Humphreys, W. Griffith
本研究旨在将质量缺陷过滤器(MDF)方法应用于使用高分辨率质谱仪筛选和鉴定活性代谢物。用高效液相色谱结合Orbitrap或傅立叶变换离子回旋共振质谱仪分析了人肝微粒体(HLM)中扑热息痛、双氯芬酸、卡马西平、氯氮平、对甲酚、4-乙基苯酚和3-甲基吲哚的谷胱甘肽(GSH)捕获的反应性代谢产物。通过选择性地去除GSH加合物MDF模板窗口外的所有离子，处理后的全扫描MS色谱图显示GSH加合物为主要组分，没有或仅有少量干扰峰。准确的质量LC MS数据集也被用于消除假阳性峰，用其他MDF模板检测稳定的氧化代谢物，以及确定代谢物分子式。与三重四极杆扫描仪中性丢失扫描相比，MDF方法在筛选HLM和大鼠胆汁中GSH捕获的反应性代谢物方面更敏感和选择性更高，在检测不承担中性丢失129Da的GSH加合物作为显著碎裂途径时更有效。GSH加合物筛选。MDF方法的能力，以及在结构鉴定中使用准确的大量MS/MS信息，使高分辨率LC-MS成为分析反应性代谢物的有用工具。
The present study was designed to apply the mass defect filter (MDF) approach to the screening and identification of reactive metabolites using high-resolution mass spectrometry. Glutathione (GSH)-trapped reactive metabolites of acetaminophen, diclofenac, carbamazepine, clozapine, p-cresol, 4-ethylphenol, and 3-methylindole in human liver microsomes (HLM) were analyzed by HPLC coupled with Orbitrap or Fourier transform ion cyclotron resonance mass spectrometry. Through the selective removal of all ions that fall outside of the GSH adduct MDF template windows, the processed full scan MS chromatograms displayed GSH adducts as major components with no or a few interference peaks. The accurate mass LCMS data sets were also utilized for the elimination of false positive peaks, detection of stable oxidative metabolites with other MDF templates, and determination of metabolite molecular formulas. Compared to the neutral loss scan by a triple quadrupole instrument, the MDF approach was more sensitive and selective in screening for GSH-trapped reactive metabolites in HLM and rat bile and far :more effective in detecting GSH adducts that do not afford the neutral loss of 129 Da as a significant fragmentation pathway. The GSH adduct screening. capability of the MDF approach, together with the utility of accurate mass MS/MS information in structural elucidation, makes high-resolution LC-MS a useful tool for analyzing reactive metabolites.