Abstract: OBJECTIVE To increase the dispersion and stability of prepared ginkgolide B(GB) in water by prepared GB liposomes. METHODS The liposome and free drug were separated by ultrafiltration centrifugation using ultrafiltration tube. The content and encapsulation efficiency of GB liposome were determined by HPLC. The method of film dispersion was used to prepare GB liposome. The influence factors on the encapsulation efficiency such as drug-to-lipid ratio, cholesterol dosage, loading amount, hydration temperature and hydration time were analyzed by single factor and orthogonal experiments. The appearance, encapsulation efficiency and particle size were used as evaluation indicators to determine the best formulation and preparation process of GB liposomes. RESULTS The best prescription for GB liposomes were as follows:10 mg GB, 160 mg egg yolk lecithin, 40 mg cholesterol, 8 mg vitamin E, 58 mg DSPE-PEG2000, 5 mL PBS buffer. The optimized process conditions were as follows:GB and lipid excipients were dissolved in absolute ethanol, and ethanol was removed by rotary evaporation under reduced pressure in a water bath at 33℃. The lipid film was hydrated with PBS buffer at pH 6.5, and the hydration temperature was 35℃, 120 min, the crude liposome suspension was ultrasonicated for 6 min in an ice water bath with an ultrasonic power of 120 W. The prepared GB liposome suspension was slightly bluish blue, light transmissive, with a particle size of (126±4)nm, an encapsulation efficiency above 87%, and no aggregation or sedimentation after 24 h. CONCLUSION GB is successfully prepared into stable GB liposomes by the thin film method in the laboratory stage. The preparation and preparation process are selected and optimized. A reference method for the formulation of GB preparations was explored and provided.