Global profiling of gut microbiota-associated lipid metabolites in antibiotic-treated mice by LC-MS/MS-based analyses

Nobuyuki Okahashi; Masahiro Ueda; Shu Yasuda; Hiroshi Tsugawa; Makoto Arita

doi:10.1016/j.xpro.2021.100492

Global profiling of gut microbiota-associated lipid metabolites in antibiotic-treated mice by LC-MS/MS-based analyses

STAR Protoc. 2021 Apr 21;2(2):100492. doi: 10.1016/j.xpro.2021.100492. eCollection 2021 Jun 18.

Authors

Nobuyuki Okahashi^{1

2}, Masahiro Ueda^{1

3}, Shu Yasuda¹, Hiroshi Tsugawa^{1

4

5}, Makoto Arita^{1

5

6}

Affiliations

¹ RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
² Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, Suita, Osaka 565-0871, Japan.
³ JSR-Keio University Medical and Chemical Innovation Center, Shinjuku-ku, Tokyo 160-0016, Japan.
⁴ RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa 230-0045, Japan.
⁵ Graduate School of Medical Life Science, Yokohama City University, Yokohama, Kanagawa 230-0045, Japan.
⁶ Division of Physiological Chemistry and Metabolism, Graduate School of Pharmaceutical Sciences, Keio University, Minato-ku, Tokyo 105-8512, Japan.

Abstract

We describe a protocol for identifying bacteria-derived lipid metabolites produced in the guts using antibiotic-treated mice, liquid chromatography tandem mass spectrometry-based lipidomics, and feature-based molecular spectrum networking (FBMN). Untargeted lipidomics using the MS-DIAL 4 program provides information on known and unknown complex lipid molecules. The FBMN technique clusters similar MS2 spectra, facilitating the identification of bacterial lipids. Targeted analysis was used as a complementary method to cover oxylipins. Here, we provide details for targeted and untargeted analyses. For complete details on the use and execution of this protocol, please refer to Yasuda et al. (2020).

Keywords: Immunology; Mass Spectrometry; Metabolomics; Microbiology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Anti-Bacterial Agents / pharmacology*
Chromatography, Liquid / methods*
Gastrointestinal Microbiome* / drug effects
Gastrointestinal Microbiome* / physiology
Lipidomics / methods*
Mice
Tandem Mass Spectrometry / methods*

Substances

Anti-Bacterial Agents