Comprehensive metabolite quantitative assay based on alternate metabolomics and lipidomics analyses
Comprehensive metabolite quantitative assay based on alternate metabolomics and lipidomics analyses
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DOI：
10.1016/j.aca.2022.339979
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发表时间：
2022-06-02
影响因子：
6.2
通讯作者：
Xu，Guowang
中科院分区：
文献类型：
--
作者：
Lv，Wangjie;Zeng，Zhongda;Xu，Guowang
基于代谢组学的精准医疗面临着跨平台数据比对问题和缺乏健康人群代谢组学基准值等障碍，其中一个主要原因是缺乏全面的代谢组学定量方法。在这里，我们开发了一种替代的反相液相色谱-质谱（RPLC-MS）方法来定量测定代谢物和脂质。在广泛的参比标准品和真实的样品的辅助下，在代谢组学和脂质组学分析中分别定义了多达397个多反应监测（MRM）跃迁（239个用于正离子模式，158个用于负离子模式）和1080个MRM跃迁（607个用于正离子模式，473个用于负离子模式），定量了超过1000种代谢物和脂质。其中，144种分析物，包括胺、氨基酸、苯丙氨酸、肽、核碱基和相关物、胆汁酸、羧酸、脂肪酸、激素、吲哚等，被绝对定量，而肉毒碱、溶血磷脂酰胆碱、溶血磷脂酰乙醇胺、游离脂肪酸、鞘磷脂、磷脂酰胆碱（PC）、烷基和烯基取代的PC、磷脂酰乙醇胺（PE）、烷基和链烯基取代的PE和三酰基甘油被半定量。所开发的方法被验证具有良好的分析特性。标准参考物质1950人血浆的分析结果与文献数据吻合良好。作为应用的证明，该方法被用于研究高尿酸血症和非酒精性脂肪肝患者的血清代谢模式的变化。这种用于定量代谢物和脂质分析的替代RPLC-MS方法可进一步用于为精准医学和生命科学提供技术和大规模数据支持。
Metabolomics-based precision medicine is facing several obstacles including cross-platform data comparison issue and the lack of metabolome benchmark values of healthy population, one of main reasons is the shortage of comprehensive metabolome quantitation methods. Here, we developed an alternate reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) method to quantitatively determine metabolites and lipids. Assisted by a wide set of reference standards and real samples, up to 397 multiple reaction monitoring (MRM) transitions (239 for positive and 158 for negative ion modes) and 1080 MRM transitions (607 for positive and 473 for negative ion modes) were defined respectively in the metabolomic and lipidomic analyses with more than 1000 metabolites and lipids being quantified. Among them, 144 analytes including amines, amino acids, benzenoids, peptides, nucleobases and related, bile acids, carboxylic acids, fatty acids, hormones, indoles and others were absolutely quantified, while carnitines, lyso-phosphatidylcholines, lyso-phosphatidylethanolamines, free fatty acids, sphingomyelins, phosphatidylcholines (PCs), alkyl and alkenyl substituted PCs, phosphatidylethanolamines (PEs), alkyl and alkenyl substituted PEs and triacylglycerols were semiquantified. The developed method was validated to have good analytical characteristics. Analytical results of standard reference material 1950 human plasma had a good agreement with literature data. As a proof of application, this method was used to study serum metabolic pattern changes of patients with hyperuricemia and nonalcoholic fatty liver. This alternate RPLC-MS method for quantitative metabolites and lipids analysis can further be used to provide technology and large-scale data support for precision medicine and life sciences.