Fatty Acid Profiling of C. elegans

Fatty acid contents were measured as previously described (Brock et al. 2006 (link)) with some modifications. 500-1,000 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° for 1 hr. Then, 1 ml supernatant was mixed with 1.2 ml hexane and 1.8 ml water to extract fatty acid methyl esters for GC-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.