*2.4. Fatty Acid Profile*

Fatty acid profile was measured according to Santos et al. [23]. Briefly, 2 mL of n-hexane was added to 0.02 g of oil to obtain fatty acid methyl esters (FAME) by cold transmethylation with methanolic potassium hydroxide. Then, 200 μL of methanolic potassium hydroxide solution (2 N) was added and vigorously mixed. Then, the supernatant was carefully transferred to a glass vial and analyzed by gas chromatography in a Shimadzu GC-2010 Plus Gas Chromatograph (Shimadzu, Tokyo, Japan). This was performed using a CPSil 88 fused silica capillary column (50 m × 0.25 mm i.d.), 0.20 μm film thickness (Varian, Middelburg, The Netherlands), and helium was used as the carrier gas (120 kPa). The used temperature program was a first step of 5 min at 140 ◦C, followed by an increase of 5 ◦C/min from 140 to 220 ◦C, and then maintaining at 220 ◦C for 15 min. The temperature of the injector and detector was 250 and 270 ◦C, respectively, and the split ratio was 1:50 with an injection volume of 1 μL. Lastly, each FAME was identified by direct comparison with a standard mixture (FAME 37, Supelco, Bellefonte, PA, USA). All analyses were performed in duplicate, and results are expressed as the relative percentage of each FA on the basis of relative peak areas.
