HPLC法测定黄藤素纳米柔性脂质体的含量及包封率

Determination of the content and encapsulation rate of palmatine-loaded flexible nano-liposomes by HPLC

目的建立黄藤素纳米柔性脂质体的含量及包封率测定方法。方法采用注入法制备黄藤素纳米柔性脂质体,用透射电镜和激光粒度仪评价脂质体的粒径和结构,鱼精蛋白凝聚法分离脂质体。选用Hypersil BDS C18(150mm×4.6mm,5.0μm)为色谱柱;流动相为乙腈-0.4mL·L-1磷酸(20∶80);检测波长:345nm;流速:1.0mL·min-1;进样量:20μL;柱温:25℃。测定黄藤素纳米柔性脂质体的含量及包封率。结果在所选定的色谱条件下,辅料及溶剂对测定无干扰,黄藤素在0.84～25.2μg·mL-1范围内线性良好(r=0.999 8),平均回收率为96.0%,精密度RSD值小于2%。结论 HPLC法可用于测定黄藤素纳米柔性脂质体的含量及包封率,该方法准确可靠、简便快速。

Objective To establish a method for the determination of the content and encapsulation rate of palmatine-loaded flexible nano-liposomes.Methods Injection method was used to prepare the iposomes.The particle size and structure of liposomes were evaluated by transmission electron microscope and laser granulometer.Protamine aggregation method was applied to separate the free drug and liposomes,and the drug concentration was determined by HPLC.The analysis was performed on a Hypersil BDS C 18 column(150 mm×4.6 mm,5.0μm),the mobile phase consisted of acetonitrile-0.4 mL·L^-1 phosphoric acid(20∶80),the detection wavelength was 345 nm,the flow rate was 1.0 mL·min^-1,the sample quantity was 20μL,and the column temperature was 25℃.Results The linear range for palmatine was 0.84-25.2μg·mL^-1(r=0.999 8).The average recovery was 96.0%with RSD<2%.Conclusion The method is simple,accurate,sensitive,and can be used for the determination of the content and encapsulation rate of palmatine-loaded flexible nano-liposomes.