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. 2023 Mar 22;8:137. doi: 10.1038/s41392-023-01380-0

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Mass spectrometry (MS)-metabolomic workflow. 1. Samples preparation consists of metabolism quenching and metabolites extraction. 2. Metabolites may need a separation step with Gas chromatography (GC), Liquid chromatography (LC), Capillary electrophoresis (CE) or can be directly ionized in the Direct infusion (DI) and in the Mass spectrometry imaging (MSI). 3. Different ionization techniques can be employed: Electron impact ionization (EI), Chemical ionization (CI), Atmospheric pressure chemical ionization (APCI), Electrospray ionization (ESI), Matrix-assisted laser desorption ionization (MALDI) and Direct real-time analysis (DART). 4. Single (MS) or tandem (MS/MS) mass analysers can be alternatively employed to separate ions according to their m/z: Quadrupole (Q), Quadrupole ion trap (QIT), Time-of-flight analyser (TOF), Fourier transform ion cyclotron resonance (FTICR), Orbitrap (OT). 5. Data processing includes conversion of m/z values, detection, filtering, normalization and identification. b Schematic depicting the most suitable techniques to separate metabolites with distinct polarity. This figure was created with Biorender.com