PMID- 34328315 OWN - NLM STAT- MEDLINE DCOM- 20210831 LR - 20210831 IS - 1520-6882 (Electronic) IS - 0003-2700 (Linking) VI - 93 IP - 31 DP - 2021 Aug 10 TI - Systematic, Modifying Group-Assisted Strategy Expanding Coverage of Metabolite Annotation in Liquid Chromatography-Mass Spectrometry-Based Nontargeted Metabolomics Studies. PG - 10916-10924 LID - 10.1021/acs.analchem.1c01715 [doi] AB - 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. FAU - Zheng, Sijia AU - Zheng S AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. AD - University of Chinese Academy of Sciences, Beijing 100049, China. FAU - Zhang, Xiuqiong AU - Zhang X AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. AD - University of Chinese Academy of Sciences, Beijing 100049, China. FAU - Li, Zaifang AU - Li Z AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. AD - University of Chinese Academy of Sciences, Beijing 100049, China. FAU - Hoene, Miriam AU - Hoene M AD - Institute for Clinical Chemistry and Pathobiochemistry, University Hospital Tuebingen, Hoppe-Seyler-Str. 3, Tuebingen 72076, Germany. FAU - Fritsche, Louise AU - Fritsche L AD - German Center for Diabetes Research (DZD), Tuebingen 72076, Germany. AD - Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum Muenchen at the University of Tuebingen, Tuebingen 72076, Germany. AD - Internal Medicine 4, University Hospital Tuebingen, Otfried-Mueller-Str. 10, Tuebingen 72076, Germany. FAU - Zheng, Fujian AU - Zheng F AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. AD - University of Chinese Academy of Sciences, Beijing 100049, China. FAU - Li, Qi AU - Li Q AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. FAU - Fritsche, Andreas AU - Fritsche A AD - German Center for Diabetes Research (DZD), Tuebingen 72076, Germany. AD - Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum Muenchen at the University of Tuebingen, Tuebingen 72076, Germany. AD - Internal Medicine 4, University Hospital Tuebingen, Otfried-Mueller-Str. 10, Tuebingen 72076, Germany. FAU - Peter, Andreas AU - Peter A AD - Institute for Clinical Chemistry and Pathobiochemistry, University Hospital Tuebingen, Hoppe-Seyler-Str. 3, Tuebingen 72076, Germany. AD - German Center for Diabetes Research (DZD), Tuebingen 72076, Germany. AD - Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum Muenchen at the University of Tuebingen, Tuebingen 72076, Germany. FAU - Lehmann, Rainer AU - Lehmann R AD - Institute for Clinical Chemistry and Pathobiochemistry, University Hospital Tuebingen, Hoppe-Seyler-Str. 3, Tuebingen 72076, Germany. AD - German Center for Diabetes Research (DZD), Tuebingen 72076, Germany. AD - Institute for Diabetes Research and Metabolic Diseases of the Helmholtz Zentrum Muenchen at the University of Tuebingen, Tuebingen 72076, Germany. FAU - Zhao, Xinjie AU - Zhao X AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. FAU - Xu, Guowang AU - Xu G AUID- ORCID: 0000-0003-4298-3554 AD - CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. AD - University of Chinese Academy of Sciences, Beijing 100049, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210730 PL - United States TA - Anal Chem JT - Analytical chemistry JID - 0370536 SB - IM MH - Chromatography, High Pressure Liquid MH - Chromatography, Liquid MH - Databases, Factual MH - Female MH - Humans MH - Mass Spectrometry MH - *Metabolomics MH - Pregnancy EDAT- 2021/07/31 06:00 MHDA- 2021/09/01 06:00 CRDT- 2021/07/30 12:11 PHST- 2021/07/31 06:00 [pubmed] PHST- 2021/09/01 06:00 [medline] PHST- 2021/07/30 12:11 [entrez] AID - 10.1021/acs.analchem.1c01715 [doi] PST - ppublish SO - Anal Chem. 2021 Aug 10;93(31):10916-10924. doi: 10.1021/acs.analchem.1c01715. Epub 2021 Jul 30.