【摘要】 目的:考察蜂毒素脂质体包封率的测定方法和游离药物的含量测定方法,对蜂毒素脂质体进行质量评价。方法:对于游离蜂毒素的含量测定,比较了单波长考马斯亮蓝法、双波长考马斯亮蓝法、福林-酚法、二喹啉甲酸法及高效液相色谱法等5种含量测定方法。对于蜂毒素脂质体包封率测定,筛选了透析法及超滤-离心法。结果:在吸收度及线性均良好的情况下,双波长考马斯亮蓝法可用于测定蜂毒素脂质体包封率时游离蜂毒素的含量,检测范围为0.5~25.0μg/mL,范围内线性良好,相关系数为0.9996。脂质体包封率测定采用超滤离心法,游离药物的平均回收率在95.75%~98.30%之间,脂质体不能透过截留相对分子质量为105的超滤膜。采用超滤离心法结合双波长考马斯亮蓝法测定3批蜂毒素脂质体的包封率为(91.45±0.83)%,RSD小于1%。结论:该方法可用于蜂毒素脂质体的质量评价。更多还原

【Abstract】 Aim:To screen an appropriate method to determine the entrapment efficiency of melittin liposomes as well as the concentration of free melittin so as to evaluate the quality of melittin liposomes. Methods:Five methods,including Bradford,two-colorimetric Coomassie brilliant blue,Lowry,BCA and HPLC were compared to determine free melittin concentration. Dialysis and ultrafiltration-centrifugation methods were compared to determine the entrapment efficiency as well. Results:As the liposomes were seized by the ultrafiltration membrane with molecular weight of 100 000,ultrafiltrationt-centrifugation method allowed separation of melittin from the liposomes,and the mean recovery of free melittin was in the range of 95.75%-98.30%. The linear range of melittin was 0.5-25.0 μg/mL (r=0.999 5). Entrapment efficiency of three batches of melittin liposomes was (91.45±0.83)% with RSD of less than 1%. Conculsion:Quality of melittin liposomes could be evaluated by the ultrafiltration-centrifugalization method.更多还原