Triglyceride Characterization in Huh-7 Cells

TG were separated from lipid extracts of Huh-7 cells by TLC in hexane/diethyl ether/acetic acid (70/30/1, v/v). The area corresponding to TG was scraped off the plates and TG was extracted from silica gel. Isolated TG were methylated with 2.5% H₂SO₄ in methanol. The resulting fatty acid methyl esters were then extracted with hexane and subjected to GC-MS analysis as described previously.38 (link) GC-MS analysis was performed by using an Agilent 7890A-5975C GC-MS network system (Agilent Technologies, Wilmington, Delaware, USA) equipped with a DB-23 capillary column (60 m × 250 μm × 0.15 μm; Agilent Technologies). The oven temperature programme was as follows; the initial temperature was 50°C for 1 min, then raised to 175°C at 25°C/min, to 235°C at 5 °C/min, and held for 5 min. The injector and detector temperatures were both set at 250°C.