Comprehensive Liver Lipid Profiling

Lipids were extracted from mouse liver tissues or Huh7 cells in 1 mL methanol containing a cohort of internal lipid standards (Avanti^® Polar Lipids, Alabaster, USA). The liver tissues isolated from the first 8 animals of each genotype were analysed. Hepatic levels of TG, CE, and phospholipids were quantified using untargeted lipidomic profiling on a Q Exactive HF-X mass spectrometer, following lipid separation on a Waters Acquity C18 UPLC column [53 ]. LipidSearch software was used for lipid annotation, chromatogram alignment and peak integration [75 (link)]. In contrast, FFAs, DG, FC, and sphingolipids were determined by targeted lipidomics on a TSQ Altis triple quadrupole mass spectrometer, following lipid separation on an Agilent Eclipse Plus C8 column [28 (link), 53 ]. Peaks were integrated using Xcalibur (Thermo Fisher, Waltham, USA) [28 (link)].