Objective To study the optimum preparation process of gastrodin liposome, characterization its structure.Methods Perpared gastrodin liposome with film-ultrasonic method.The entrapment efficiency as index, quality between lecithin and cholesterol ratio, oil-water phase volume ratio and ultrasonic time as factors by L9 (34 ) orthogonal test to optimize the preparation process.Observed the liposome morphology, particle size and distribution under the microscope, and determined the entrapment efficiency and stability.Results Regression equation was Y=239.75 +0.0207X(n=5), R2 =0.9996, 0.002 to 0.010 mg/mL had a good linear relationship.The optimum of preparation process: lecithin and cholesterol quality ratio was 4:1, water and oil phase volume ratio of 3:1, ultrasonic time control for 60 min;Liposomes showed spherical or oval shape, regular shape, uniform size.The average entrapment efficiency was 60.65%, the average particle size distribution at 327 nm, and it had good stability.Conclusion The preparation of gastrodine liposome has advantages of simple, higher entrapment efficiency and even particle size.The method can be used for embedded water-soluble medicine.