the samples (1g) were saponified with 1 N KOH in ethanol (20 ml) in a water bath (80) for 1 h
after cooling, 10 ml of each sample was transferred to 50-ml tubes,vortexed with 6N HCL (3ml) and hexane (10ml) for the extraction of fatty acid and then evaporated by nitrogen gas (99.99%)