A Systematic Pipeline to Enhance the Fecal Metabolome Coverage by LC-HRMS

Journal of the Brazilian Chemical Society

article

Alves, Marina A.¹  Silva, Ana Carolina R. da¹  Torres, Clarisse L.¹  Almeida, Lana R. de³  Mastella, Ana Maria O.³  Borges, Ricardo M.⁵  Garrett, Rafael¹ 

[1]Laboratório de Metabolômica (LabMeta, Instituto de Química, Universidade Federal do Rio de Janeiro (UFRJ)

[2]Turku Bioscience Centre, University of Turku and Åbo Akademi University

[3]Laboratório de Evolução, Sistemática e Ecologia de Aves e Mamíferos (BiMaLab), Universidade Federal do Rio Grande do Sul

[4]Programa de Pós-Graduação em Biologia Animal, Universidade Federal do Rio Grande do Sul

[5]Instituto de Pesquisas de Produtos Naturais Walter Mors (IPPN), Universidade Federal do Rio de Janeiro

关键词: LC-HRMS;    metabolomics;    lipidomics;    fecal;    molecular networking;   

DOI  :  10.21577/0103-5053.20210042

学科分类：内科医学

来源: SciELO

【 摘 要 】

The comprehended knowledge of the metabolic profile of the fecal matter has been recognized as an important point for understanding metabolic changes in the human systemic metabolism and it can provide precious information about host-gut microbiota interactions. However, few analytical strategies have been addressed for a broad analysis of metabolites with different chemical properties to better understand the chemical space of fecal samples. Here we report a systematic pipeline to achieve comprehensive coverage of the fecal metabolome, from high polar to nonpolar metabolites, using dog fecal samples as a proof-of-concept. This pipeline comprises a monophasic (ACN/H 2 O) and a biphasic extraction (methyl tert- butyl ether (MTBE)/MeOH/H 2 O) of the sample, followed by three liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) methods using HILIC-amide, RP-C 18 and CSH-C 18 columns, and a switch polarity acquisition mode in the electrospray ion source. This approach allowed the annotation of 376 metabolites from 70 different chemical classes. The chemical space analysis by molecular networking and the pathway analysis revealed the complexity of the fecal sample and the importance of combined methods to better understand biochemical pathways. This pipeline can be used as a valuable tool to comprehend the relationship between host-gut microbiota metabolites and the influence of diet, medication, or environmental changes.

【 授权许可】

CC BY