Abstract: This experiment prepared high similarity human milk substitute fats (HMFS) from different sources and analyzed their differences in oxidative stability. The Fmincon Toolbox of MATLAB software was utilized to optimize four different sources of HMFS. Animal and vegetable oils with different sources and special composition characteristics as well as commercial OPO products, have been selected as the base oils for blending. The content of major fatty acids (FA), sn-2 FA and triglycerides (TAG) in Chinese HMF was used as the blending target. The acid value (AV), peroxide value (PV), anisidine value (p-AV) and total oxidation value (TOTOX) of HMFS during accelerated oxidation were also monitored by oven accelerated oxidation test. Through the similarity evaluation analysis of HMFS and HMF, the total similarity scores between buttermilk/vegetable oil-based HMFS (HMFS-1), goat milk/vegetable oil-based HMFS (HMFS-2), lard/vegetable oil-based HMFS (HMFS-3) and vegetable oil-based HMFS (HMFS-4) and Chinese HMF were 81.91, 83.43, 83.18, and 66.65, respectively. During the accelerated oxidation test in the oven, the AV of the four HMFS showed a slow increasing trend over time, ultimately all below 1 mg/kg. The trends of PV, p-AV, and TOTOX of animal oil-based HMFS were similar, ranging from (0.18±0.02) to (0.32±0.01) mmol/kg, 1.6±0.0 to 3.5±0.1, and 2.93±0.12 to 4.65±0.05 on the 21st day, respectively. The oxidation degree of vegetable oil-based HMFS was the highest during the accelerated oxidation process, and the increase in the three oxidation indicators was always higher than that of animal/vegetable oil-based HMFS, ultimately reached (1.36±0.01) mmol/kg, 22.7±0.2, and 28.13±0.01, respectively. Compared to vegetable oil-based HMFS, animal/vegetable oil-based HMFS had higher similarity and stronger oxidative stability with human milk fat. Therefore, adding milk fat or lard to HMFS is an effective method to increase its similarity with human milk fat and enhance oxidative stability.