Pseudotargeted Method based on Parallel Column Two-Dimensional Liquid Chromatography-Mass Spectrometry for Broad Coverage of Metabolome and Lipidome.
Pseudotargeted Method based on Parallel Column Two-Dimensional Liquid Chromatography-Mass Spectrometry for Broad Coverage of Metabolome and Lipidome.
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DOI：
10.1021/acs.analchem.0c00372
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发表时间：
2020-03
影响因子：
7.4
通讯作者：
Wangjie Lv;Lichao Wang;Qiuhui Xuan;Xinjie Zhao;Xinyu Liu;Xianzhe Shi;Guowang Xu
中科院分区：
文献类型：
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作者：
Wangjie Lv;Lichao Wang;Qiuhui Xuan;Xinjie Zhao;Xinyu Liu;Xianzhe Shi;Guowang Xu
由于极性差异较大，代谢物和脂质分析通常需要两种液相色谱 - 质谱联用（LC - MS）方法。拟靶向代谢组学方法作为一种融合了非靶向和靶向方法优势的新途径应运而生。在此，我们旨在通过基于平行柱的二维液相色谱（PC - 2DLC） - 质谱联用技术和拟靶向方法，建立一种针对代谢组和脂质组的综合分析方法。为了尽可能同时提取多种极性代谢物和非极性脂质，我们系统地优化了样品前处理过程，分别选用异丙醇/甲醇（3:1，v/v）和异丙醇/水（7:3，v/v）作为提取溶剂和复溶溶剂。样品前处理优化后，检测到的甘油三酯显著增加。随后，采用PC - 2DLC与三重四极杆飞行时间质谱（Triple TOF MS）联用，对来自血清、尿液和肝组织基质的混合样品进行分析。分别依据“MRM离子对查找器”软件和脂质组学MRM转换数据库，确定代谢组和脂质组的多反应监测（MRM）转换。经QTRAP MS在预定MRM模式下验证后，在正离子模式和负离子模式下分别检测到对应1294个MRM转换的1609种潜在代谢物和脂质，以及对应687个MRM转换的847种潜在代谢物和脂质。它们的辛醇/水分配系数跨度约30个数量级。验证表明拟靶向2DLC - MS方法具有良好的分析特性。作为适用性验证，利用所建立的方法对2型糖尿病患者的血清进行了研究。结果表明，拟靶向2DLC - MS方法可靠、可重复，可用于代谢组学研究。
Metabolite and lipid profilings usually need two liquid chromatography-mass spectrometry (LC-MS) methods due to great polarity difference. Pseudotargeted metabolomics method has emerged as a novel approach integrating the advantages of nontargeted and targeted methods. Here, we aim to establish a comprehensive method for metabolome and lipidome by using parallel column-based two-dimensional LC (PC-2DLC)-MS and pseudotargeted approach. To simultaneously extract as many polar metabolites and nonpolar lipids as possible, we systematically optimized the sample pretreatment process, and isopropanol/methanol (3:1, v/v) and isopropanol/water (7:3, v/v) were selected as the extraction and reconstitution solvents, respectively. The detected triglycerides have significantly increased after the sample pretreatment optimization. Then, PC-2DLC coupled with Triple TOF MS was applied to analyze a mixed sample from serum, urine and liver tissue matrices. The multiple reaction monitoring (MRM) transitions of metabolome and lipidome were defined according to the "MRM-Ion Pair Finder" software and lipidomics MRM-transition database, respectively. After verified by QTRAP MS in the scheduled MRM mode, 1609 potential metabolites and lipids corresponding to 1294 MRM transitions, and 847 potential metabolites and lipids corresponding to 687 MRM transitions were detected in positive and negative ion modes, respectively. They range about 30 orders of magnitude in octanol/water partition coefficient. Pseudotargeted 2DLC-MS method was validated to have good analytical characteristics. As a proof of applicability, sera from Type 2 diabetic patients were investigated by the established method. The results indicated that the pseudotargeted 2DLC-MS method is reliable, repeatable and can be used in the metabolomics study.