Overview of a mass spectrometry-based bulk lipidomics set-up. Lipids are extracted from homogenized biological samples by either liquid–liquid extraction or solid phase extraction. Crude lipid extracts can be directly injected into the mass spectrometer (‘shotgun lipidomics’) or can be separated by LC first. ESI is the preferred mode of ionization for the vast majority of lipids, while the choice of mass analyser often depends on whether the experiment is targeted or untargeted (discussed in the text). (This image was created with BioRender.com.)