Fatty acid methyl ester (FAME) content analysis were weighed into clean, 20 ml screw-top glass tubes, to which 4 ml fresh solution of a mixture of methanol, concentrated sulfuric acid, and chloroform (17:0.3:2.0 v/v/v) was added. The tubes were closed tightly with Teflon tape to transesterification, tubes were avoid leakag e, and then weighed. For placed inside a heating block at temperatures of 90°C and heated for 60 min. On completion of the reaction, the tubes were cooled down to room temperature and weighed again to dismiss leaking samples Then, 1 ml distilled water was added into the mixture, and the mixture was thoroughly vortexed for 1 min. After the formation of two phases, the lower phase containing FAME was transferred to a 1.5 ml GC vial. samples were stored in the freezer (-20°C) until GC-Ms analysis [10]