The combination of thin film hydration and ultrasonic treatment was used to prepare folic acid liposomes with lecithin and cholesterol as the membrane material and folic acid as the core material. Dialysis-spectrophotometry method was applied to determine the encapsulation efficiency of folic acid liposomes. Based on a series of single-factor experiments, the optimal conditions for preparing folic acid liposomes were explored by orthogonal array design to be preparation temperature of 60 ��, pH 8.0, the mass ratio between lecithin and cholesterol of 5:1, and folic acid concentration of 1.4 mg/mL. The prepared liposomes under these optimal conditions had an entrapment efficiency of 30.36% with an average particle size of 335.4 nm. The liposomal suspension was sealed in a bottle and stored at 4 �� for 15 days for stability evaluation based on measurement of average particle size, and the results indicated that the average particle size had no significant change, which was in the range of 300 to 400 nm, and that the suspension had a good stability.