Abstract: Leaching method, Soxhlet method and ultrasonic-assisted method were selected to obtain oil from Chlorella vulgaris C9-JN2010. Both the effect of extraction time on extraction ratio and the effect of extraction methods on fatty acids composition were studied after the best slovent of each method were selected. The results showed that the extraction ratio of leaching method, Soxhlet extraction method and ultrasonic-assisted extraction method could come up to 84.06%, 89.11% and 88.66%, respectively. And the lipid obtained by those three methods was not significantly different in fatty acids composition. As the most efficiency method, ultrasonic-assisted method was suitable for industrial amplification. The oil from Chlorella vulgaris C9-JN2010 had potential to be functional oil because of the high content of unsaturated fatty acid, such as linolenic acid and linoleic acid.