2.7.7. Fatty Acid Methyl Ester (FAME) Analysis by GC–MS

FAMEs were analysed by gas chromatography–mass spectrometry (GC–MS) consisting of a Varian 3800 GC and a Bruker 300 triple quadrupole MS fitted with a 30 m × 0.25 mm Agilent VF-5MS column (0.25 µm film thickness, bound fused silica, Agilent Technologies, Santa Clara, CA, USA) and using helium (1.3 mL/min) as the carrier gas. A 1 µL portion of sample was injected with a 30:1 split ratio. The injection port was kept at 290 °C. The oven temperature was initially 50 °C for 1 min and then programmed to heat at a rate of 30 °C/min to reach 150 °C, then 2 °C/min to reach 250 °C and after that, 5 °C/min to reach the final temperature 320 °C for 5 min. Star software was used to control the operation of the GC–MS. Full-scan MS spectra in the range of (m/z) 35 to 450 was obtained. Individual FAMEs were identified by comparison of retention times and mass spectra with those of standard FAME (NIST/EPA/NIH Mass Spectral Library 2017). Relative amounts of each FAME were calculated as a percentage of the total amount (peak area) detected.