Coupling Targeted and Untargeted Mass Spectrometry for Metabolome-Microbiome-Wide Association Studies of Human Fecal Samples
Coupling Targeted and Untargeted Mass Spectrometry for Metabolome-Microbiome-Wide Association Studies of Human Fecal Samples
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DOI：
10.1021/acs.analchem.7b01381
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发表时间：
2017-07-18
影响因子：
7.4
通讯作者：
Dorrestein, Pieter C.
中科院分区：
文献类型：
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作者：
Melnik, Alexey V.;da Silva, Ricardo R.;Dorrestein, Pieter C.
人们越来越认识到肠道微生物组在健康中的作用，这有助于了解人类粪便的分子组成。为了分析此类复杂的样品，我们开发了一个耦合靶向和非靶向代谢组学的平台。该方法通过来自一个 UPLC 的分流、由触点闭合继电器触发的联合定时以及用于检索数据的脚本来实现。它旨在以高灵敏度检测感兴趣的特定代谢物，允许校正目标信息，实现更好的定量，从而为探索性研究提供先进的分析工具。 Procrustes 分析表明，非靶向方法提供了与微生物组数据更好的相关性，将特定代谢物与产生或处理它们的微生物联系起来。对于来自 American Gut 项目的一百多个人类粪便样本的子集，所描述的耦合工作流程的实施表明，使用每种化合物的单个转变与保留时间相结合的针对性分析会错误识别 30% 的目标数据，并可能导致不正确的解释。同时，针对性分析将检测限和动态范围扩展了几个数量级，具体取决于化合物。软件应用程序已开发为工作流程的一部分，可根据校准曲线进行定量评估。使用这种方法，我们检测了粪便中预期的微生物修饰分子（例如次级胆汁酸）和意想不到的微生物分子（包括假单胞菌相关的喹诺酮类药物和鼠李糖脂），为代谢组微生物组范围内的关联研究（MMWAS）奠定了基础。
Increasing appreciation of the gut microbiome's role in health motivates understanding the molecular composition of human feces. To analyze such complex samples, we developed a platform coupling targeted and untargeted metabolomics. The approach is facilitated through split flow from one UPLC, joint timing triggered by contact closure relays, and a script to retrieve the data. It is designed to detect specific metabolites of interest with high sensitivity, allows for correction of targeted information, enables better quantitation thus providing an advanced analytical tool for exploratory studies. Procrustes analysis revealed that untargeted approach provides a better correlation to microbiome data, associating specific metabolites with microbes that produce or process them. With the subset of over one hundred human fecal samples from the American Gut project, the implementation of the described coupled workflow revealed that targeted analysis using combination of single transition per compound with retention time misidentifies 30% of the targeted data and could lead to incorrect interpretations. At the same time, the targeted analysis extends detection limits and dynamic range, depending on the compounds, by orders of magnitude. A software application has been developed as a part of the workflow to allows for quantitative assessments based on calibration curves. Using this approach, we detect expected microbially modified molecules such as secondary bile acids and unexpected microbial molecules including Pseudomonas-associated quinolones and rhamnolipids in feces, setting the stage for metabolomemicrobiome-wide association studies (MMWAS).