To analyze the fatty acid composition, fat was extracted according
to the method of Folch et al [18], and 2 mL 0.5N NaOH/
methanol was added to 20 mg of fat, which was later saponified for
10 minutes at 105C. It was examined after applying 2 mL boron
trifluoride/methanol, and methylated. Then, 2e3 mL hexane (HPLC
grade) and 2 mL saturated NaCl solution were added. The supernatant
of the mixture used the separated funnel was analyzed by
gas chromatography (Hewlett Packard 6890 series; Palo Alto, CA,
USA). The column was set up with an HP-FFAP capillary column
(25 m 0.32 mm internal diameter, 0.5 mm film thickness); initial
oven temperature of 130C (1 minute), increased at 2.5C/min to a
final temperature of 230C (10 minutes); injector temperature
230C, detector temperature 250C; helium carrier gas with a spilt
ratio of 20 : 1, and flow rate of 1 mL/min.