Ion-Pair Selection Method for Pseudotargeted Metabolomics Based on SWATH MS Acquisition and Its Application in Differential Metabolite Discovery of Type 2 Diabetes
Ion-Pair Selection Method for Pseudotargeted Metabolomics Based on SWATH MS Acquisition and Its Application in Differential Metabolite Discovery of Type 2 Diabetes
复制标题
基于SWATH MS采集的伪靶向代谢组学离子对选择方法及其在2型糖尿病差异代谢物发现中的应用
DOI：
10.1021/acs.analchem.8b02377
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发表时间：
2018-10-02
影响因子：
7.4
通讯作者：
Xu, Guowang
中科院分区：
文献类型：
--
作者：
Wang, Lichao;Su, Benzhe;Xu, Guowang
伪靶向代谢组学方法结合了非靶向和靶向分析的优点，因为它可以在多反应监测(MRM)模式下获得代谢产物的数据，而不需要标准。关键是从待分析样品中收集离子对信息。众所周知，所有理论碎片离子(SWATH)MS模式的顺序加窗采集可以最大程度地获取MS2信息。为了以最佳的碰撞能量(CE)方便地获取尽可能多的离子对，提出了一种基于可变隔离窗口的SWATH MS采集的离子对选择方法。首先，利用超高效液相色谱-四极杆飞行时间(Q-TOF)MS平台和3个CE对血浆标准物质(SRM 1950)中的所有代谢物信息进行非靶向采集。借助于软件工具，获得了独特代谢物的离子对。然后在计划的MRM和UHPLC的耦合中对它们进行验证。在去除假阳性后，具有最佳CE的离子对被积分。在正离子模式下，共获得了第1373个独特的代谢物离子对。并通过日内和日间精密度评价所建立的假靶向方法的可重复性。结果表明，该方法稳定、可靠，适用于代谢组学研究。作为应用实例，应用所建立的方法研究了2型糖尿病患者血清代谢物的变化。本工作提供了一种基于SWATH MS采集的伪靶向离子对选择方法，具有代谢物覆盖率高、毛细管电泳法合适、加工方便等特点。
The pseudotargeted metabolomics method integrates advantages of nontargeted and targeted analysis because it can acquire data of metabolites in the multireaction monitoring (MRM) mode of mass spectrometry (MS) without needing standards. The key is the ion-pair information collection from samples to be analyzed. It is well-known that sequential windowed acquisition of all theoretical Fragment ion (SWATH) MS mode can acquire MS2 information to a maximum extent. To expediently acquire as many ion-pairs as possible with optimal collision energy (CE), an ion-pair selection approach based on SWATH MS acquisition with variable isolation windows was developed in this study. Initially, nontargeted acquisition of all metabolites information in plasma Standard Reference Material (SRM 1950) was performed by ultra high-performance liquid chromatography (UHPLC)-quadrupole time-of-flight (Q-TOF) MS platform with three CEs. With the help of software tool, the ion-pairs of unique metabolites were gained. Then they were validated in scheduled MRM coupled with UHPLC. After removing false positive, the ion-pairs with an optimal CE was integrated. A total of 1373 unique metabolite ion-pairs were obtained at positive ion mode. And repeatability of the established pseudotargeted approach was evaluated by intraday and interday precision. The results demonstrated the method was stable, reliable, and suitable for metabolomics study. As an application example, alterations of serum metabolites in Type 2 diabetes were investigated by using the established method. This work provides a pseudotargeted ion-pair selection method based on SWATH MS acquisition with the characters of increased metabolite coverage, suitable CE, and convenient processing.