Abstract: With Korean pine seed oil we enriched and purifiedpinoleic acid by urea inclusion. Through the influence of the ratio of fatty acid to urea, the ratio of urea to ethanol, we optimized the inclusion temperature and the inclusion time on the content of pinoleic acid and recovery of pinoleic acid, the process parameters of urea bagging. Finally, the optimal purification condition of pinoleic acid was determined as follows:the ratio of fatty acid to urea was 1:6.5(w/w); the ratio of urea to 90% ethanol was 1:4(w/v); Inclusion temperature was -10℃; Inclusion time was 12 h. Under the above conditions, a purity of 58.3% pinoleic acid could be obtained, and the recovery rate was 51.8%. After purification by secondary urea inclusion, a purity of 72.5% of pinoleic acid could be obtained, and the recovery rate was 64.7%. To verify the lipid-lowering activity of pinoleic acid, oleic acid was used to induce lipogenesis in HepG2 cells. The results showed 40 μmol·L^-1 pinoleic acid could significantly reduce the level of triglycerides and total cholesterol in HepG2 cells.