Xiong Yang, Guo Dan, Zheng Xiao-Li, Sun Peng, Xu Lian-Ying, Chen Jian-Ming
College of Pharmacy, the Second Military Medical University, Shanghai 200433, China.
Yao Xue Xue Bao. 2008 Feb;43(2):214-20.
Nobiliside A (Nob A) liposomes were prepared. Its assay method of content and encapsulation efficiency (EE) were established, and hemolytic activity with Nob A solution in vivo and in vitro were compared. Preparative method, phospholipid content, ratio of phospholipid to cholesterol and ratio of drug to lipids were optimized by single factor exploration. According to the optimized results, 3 batches of Nob A liposomes were prepared, then high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) method was used to determine the content of Nob A and minicolumn centrifugation method to determine EE, transmission electron microscope was used to detect the morphology and laser scatter analysis to evaluate particle sizes of the liposomes. The hemolytic activity was studied both in vivo and in vitro. The results indicated that HPLC-ELSD method and minicolumn centrifugation method used in this study are simple, applicable and accurate for the determination of the content and EE of Nob A liposome respectively . Nob A liposomes have a high EE with spherical shape and uniform size by using the film ultrasonication technique. When the ratio of phospholipid to cholesterol was 2:1 and the ratio of Nob A to lipids was 1:40, the mean EE of Nob A liposomes was 95.7% and the mean diameter was 87.6 nm. Liposomes inhibited the hemolytic activity of Nob A in vivo and in vitro sharply. As for its low hemolytic activity in vivo and in vitro, Nob A liposomes are optimistic to be used by intravenous injection.
制备了诺必苷A（Nob A）脂质体。建立了其含量及包封率（EE）的测定方法，并比较了Nob A溶液在体内和体外的溶血活性。通过单因素探索对制备方法、磷脂含量、磷脂与胆固醇的比例以及药物与脂质的比例进行了优化。根据优化结果，制备了3批Nob A脂质体，然后采用高效液相色谱-蒸发光散射检测器（HPLC-ELSD）法测定Nob A的含量，采用微柱离心法测定EE，用透射电子显微镜检测脂质体的形态，用激光散射分析评估脂质体的粒径。对溶血活性进行了体内和体外研究。结果表明，本研究中使用的HPLC-ELSD法和微柱离心法分别用于测定Nob A脂质体的含量和EE，方法简便、适用且准确。采用薄膜超声技术制备的Nob A脂质体具有较高的EE，呈球形且大小均匀。当磷脂与胆固醇的比例为2:1，Nob A与脂质的比例为1:40时，Nob A脂质体的平均EE为95.7%，平均直径为87.6 nm。脂质体在体内和体外均能显著抑制Nob A的溶血活性。由于其在体内和体外的溶血活性较低，Nob A脂质体有望用于静脉注射。
