The invention discloses a method for preparing 1, 3-dilinolein-2-palmitic acid triglyceride. The method includes: melting the catfish oil at the temperature of 50-60 DEGC for 30-60 min, reducing the temperature to 15-30 DEG C at the cooling rate of 1-5 DEG C/min for 5-48 h, eliminating the liquid oil through the filtering mode and obtaining the catfish oil solid lipid which is rich in palmitic acid; and taking sn-1, 3-digit specificity lipase as the catalyst and the free fatty acid which comes from high oleic acid vegetable oil as the acyl donor, conducting acidolysis on the catfish oil solid lipid, eliminating the free fatty acid and obtaining the product which is rich in 1, 3-dilinolein-2-palmitic acid triglyceride. The 1, 3-dilinolein-2-palmitic acid triglyceride is low in price of raw materials, high in content and capable of being applied to infant formula milk as the additive.