Abstract: The alcoholysis of oils and fats combined with urea inclusion method was used to enrich and purify pinolenic acid from the Pinus koraiensis seed oil for the first time. On the basis of investigating the influence of the ratio of P. koraiensis seed oil fatty acid ethyl esters to urea, ratio of ethanol to urea, inclusion temperature and inclusion time on the GC content of pinolenic acid ethyl ester, the enrichment and purification conditions of urea inclusion were optimized by response surface methodology. The results showed that the best purification conditions of pinolenic acid ethyl ester were the ratio of urea to P. koraiensis seed oil fatty acid ethyl esters 3:1, the ratio of ethanol to urea 6:1(mL:g), the inclusion temperature -13 ℃ and the inclusion time 24 h.Under the above optimal conditions, the purity of pinolenic acid ethyl ester could reach 94.75%, and the recovery rate could reach 66.25%. Compared with the traditional method, the purity increased by 107%, and the recovery rate increased by 143%. In addition, urea showed excellent reusability in the recovery experiment. The average purity of pinolenic acid could still reach 85.28% after three cycles of urea inclusion and recovery.