2.3.3. Lipid Extraction

Lipids were extracted immediately after digestion using hexane/isopropanol (2:1, v/v). BHT was added to the extraction solvent at a concentration of 0.05% in order to prevent further oxidations.²⁸ Throughout the procedure, samples were kept on ice. The solvent was added to the digestates in a ratio of 1:1 v/v, and tubes were vortexed for 10 s and centrifuged at 1000 rpm for 3 min. The extraction was performed twice. The upper phases were combined and evaporated to dryness with N₂ at 37 °C. Samples were recovered with 3 mL of chloroform/methanol (1:1, v/v) and stored at −80 °C.