Center for Analytical Chemistry, Department of Agrobiotechnology, IFA-Tulln, University of Natural Resources and Life Sciences, Vienna , 1180 Vienna, Austria.
Department of Sustainable Crop Production, School of Agriculture, Università Cattolica del Sacro Cuore , 29100 Piacenza, Italy.
Anal Chem. 2017 Sep 5;89(17):9518-9526. doi: 10.1021/acs.analchem.7b02518. Epub 2017 Aug 22.
Stable isotope labeling (SIL) techniques have the potential to enhance different aspects of liquid chromatography-high-resolution mass spectrometry (LC-HRMS)-based untargeted metabolomics methods including metabolite detection, annotation of unknown metabolites, and comparative quantification. In this work, we present MetExtract II, a software toolbox for detection of biologically derived compounds. It exploits SIL-specific isotope patterns and elution profiles in LC-HRMS(/MS) data. The toolbox consists of three complementary modules: M1 (AllExtract) uses mixtures of uniformly highly isotope-enriched and native biological samples for selective detection of the entire accessible metabolome. M2 (TracExtract) is particularly suited to probe the metabolism of endogenous or exogenous secondary metabolites and facilitates the untargeted screening of tracer derivatives from concurrently metabolized native and uniformly labeled tracer substances. With M3 (FragExtract), tandem mass spectrometry (MS/MS) fragments of corresponding native and uniformly labeled ions are evaluated and automatically assigned with putative sum formulas. Generated results can be graphically illustrated and exported as a comprehensive data matrix that contains all detected pairs of native and labeled metabolite ions that can be used for database queries, metabolome-wide internal standardization, and statistical analysis. The software, associated documentation, and sample data sets are freely available for noncommercial use at http://metabolomics-ifa.boku.ac.at/metextractII .
稳定同位素标记 (SIL) 技术有可能增强基于液相色谱-高分辨率质谱 (LC-HRMS) 的非靶向代谢组学方法的多个方面，包括代谢物检测、未知代谢物的注释和比较定量。在这项工作中，我们提出了 MetExtract II，这是一种用于检测生物衍生化合物的软件工具包。它利用了 LC-HRMS(/MS) 数据中 SIL 特有的同位素模式和洗脱轮廓。该工具包由三个互补的模块组成：M1（AllExtract）使用均匀高度同位素富集和天然生物样品的混合物，选择性地检测整个可及的代谢组。M2（TracExtract）特别适合探测内源性或外源性次生代谢物的代谢，并有助于从同时代谢的天然和均匀标记示踪剂中靶向筛选示踪衍生物。利用 M3（FragExtract），评估相应的天然和均匀标记离子的串联质谱 (MS/MS) 片段，并自动分配可能的总和公式。生成的结果可以以图形方式说明，并以包含所有检测到的天然和标记代谢物离子对的综合数据矩阵导出，这些离子对可用于数据库查询、代谢组内标准化和统计分析。该软件、相关文档和示例数据集可在 http://metabolomics-ifa.boku.ac.at/metextractII 免费用于非商业用途。
