银杏内酯B脂质体的处方与制备工艺研究

Study on Formulation and Preparation Process of Ginkgolide B Liposome

关键词:  银杏内酯B  脂质体  超滤离心-HPLC法  薄膜分散法

Key words:  ginkgolide B  liposome  ultrafiltration centrifugation-HPLC  film dispersion method

摘要:

目的 将银杏内酯B（ginkgolide B，GB）制备成脂质体，增加GB在水中的分散性和稳定性。方法 采用超滤离心法分离脂质体与游离药物，HPLC测定脂质体中GB的含量与包封率，采用薄膜分散法制备GB脂质体；通过单因素与正交试验分析药脂比、胆固醇用量、装载量、水化温度和水化时间等因素对包封率的影响，以外观、包封率、粒径为评价指标确定GB脂质体的最佳处方和制备工艺。结果 GB脂质体的最佳处方为10 mg GB、160 mg蛋黄卵磷脂、40 mg胆固醇、8 mg维生素E、58 mg DSPE-PEG2000、5 mL PBS缓冲液；优化后的工艺条件为GB与脂质辅料溶于无水乙醇中，33℃水浴条件下减压旋转蒸发除乙醇，pH 6.5 PBS缓冲液水化脂质薄膜，水化温度35℃，时间120 min，粗脂质体混悬液冰水浴超声6 min，超声功率120 W。制备的GB脂质体混悬液微泛蓝色乳光、可透光，粒径为（126±4） nm，包封率>87%，放置24 h无聚集、无沉降。结论 采用薄膜法在实验室阶段成功制备了稳定的GB脂质体，确定了最优制备处方和制备工艺，为GB制剂的剂型研究做出探索和提供参考。

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.