Fatty Acid Profiling in C. elegans

Fatty acid contents were measured as previously described^{65 (link)} with some modifications. About 500–1000 age-synchronized day-1 adult worms were washed off plates and washed three times with water. Worm pellets were resuspended with 1.2 mL of 2.5% H₂SO₄ in methanol and incubated at 80 °C for 1 h. Then, 1 ml of supernatant was mixed with 1.2 ml of hexane and 1.8 ml of water to extract fatty acid methyl esters (FAMEs) for GC–MS/MS analysis. The Supelco 37 Component FAME Mix (Sigma) was used to determine the retention time. The Shimadzu GCMS-TQ8040 Gas Chromatograph Mass Spectrometer equipped with SH-Rxi-5sil MS column was used. The contents of fatty acids were normalized to protein concentrations. The data were collected by GCMSsolution software.