Fish oils have been recognized as good sources of Polyunsaturated Fatty Acids (PUFA), which are widely used for pharmaceutical purpose and food supplement. In this study, the multi-objective optimization method was used to describe the enrichment process of DHA, EPA, omega-3 from pangasius fish oil with the urea-to-fish oil ratio, complexation temperature and urea complexation time which affected the enrichment’s level of polyunsaturated fatty acid. The multi-objective optimization result has determined the technological mode of enrichment process at the urea-to-fish oil ratio of 2/1, complexation temperature of -11°C and urea complexation time of 18.6 h. Under these conditions, the enrichment efficiency of DHA, EPA and omega-3 reached to maximum level of 120.06, 58.4 and 179.5% and it was the basis to establish the technology mode for the enrichment process from pangasius fish oil. The result was totally compatible with experiment and able to apply in industrial production in order to create the fish oil which contented high polyunsaturated fatty acid and better in quality.