Systematic, Modifying Group-Assisted Strategy Expanding Coverage of Metabolite Annotation in Liquid Chromatography-Mass Spectrometry-Based Nontargeted Metabolomics Studies
Systematic, Modifying Group-Assisted Strategy Expanding Coverage of Metabolite Annotation in Liquid Chromatography-Mass Spectrometry-Based Nontargeted Metabolomics Studies
复制标题
系统性、修改组辅助策略扩大基于液相色谱-质谱法的非靶向代谢组学研究中代谢物注释的覆盖范围
DOI：
10.1021/acs.analchem.1c01715
复制
发表时间：
2021
影响因子：
7.4
通讯作者：
Xu Guowang
中科院分区：
文献类型：
--
作者：
Zheng Sijia;Zhang Xiuqiong;Li Zaifang;Hoene Miriam;Fritsche Louise;Zheng Fujian;Li Qi;Fritsche Andreas;Peter Andreas;Lehmann Rainer;Zhao Xinjie;Xu Guowang
从微生物到人类，液相色谱-质谱法（MS）的非靶向代谢谱分析已被广泛用于研究代谢变化。然而，一个主要的挑战是从数千个检测到的特征中标注代谢物。我们的研究目的是通过一种集成的多步骤策略，应用基于数据依赖采集（DDA）的超高效液相色谱(UHPLC) -高分辨率质谱（HRMS）分析，然后对特征代谢物修饰进行全面的中性损失匹配，并以连续的方式进行数据库搜索，从而超越代谢物注释在常见非靶向代谢组学研究中的覆盖范围。使用汇集的人类尿液作为模型样本进行方法建立，我们发现22%的检测到的化合物具有修饰结构。尿液中代谢物改变的主要类型是葡萄糖醛酸化（33%）、磺化（20%）和乙酰化（6%）。在383个被注释的代谢物中，100个被标准化合物确认，50个在人类代谢物数据库（HMDB）和京都基因与基因组百科全书（KEGG）等常用数据库中不存在的修饰代谢物被结构解析。通过对诊断为妊娠期糖尿病的孕妇与健康对照者尿液的调查，检验了该方法的实用性。总体而言，83种差异代谢物被注释，其中67%是修饰代谢物，包括5种以前未报道的化合物。综上所述，系统修饰群辅助策略可以作为扩展生物学和生物医学非靶向研究中注释代谢物数量的有用工具。
From microbes to human beings, nontargeted metabolic profiling by liquid chromatography (LC)–mass spectrometry (MS) has been commonly used to investigate metabolic alterations. Still, a major challenge is the annotation of metabolites from thousands of detected features. The aim of our research was to go beyond coverage of metabolite annotation in common nontargeted metabolomics studies by an integrated multistep strategy applying data-dependent acquisition (DDA)-based ultrahigh-performance liquid chromatography (UHPLC)–high-resolution mass spectrometry (HRMS) analysis followed by comprehensive neutral loss matches for characteristic metabolite modifications and database searches in a successive manner. Using pooled human urine as a model sample for method establishment, we found 22% of the detected compounds having modifying structures. Major types of metabolite modifications in urine were glucuronidation (33%), sulfation (20%), and acetylation (6%). Among the 383 annotated metabolites, 100 were confirmed by standard compounds and 50 modified metabolites not present in common databases such as human metabolite database (HMDB) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were structurally elucidated. Practicability was tested by the investigation of urines from pregnant women diagnosed with gestational diabetes mellitus vs healthy controls. Overall, 83 differential metabolites were annotated and 67% of them were modified metabolites including five previously unreported compounds. To conclude, the systematic modifying group-assisted strategy can be taken as a useful tool to extend the number of annotated metabolites in biological and biomedical nontargeted studies.