超滤法-HPLC法测定灯盏花素脂质体包封率

Determination of entrapment efficiency for liposomal formulation of breviscapine using ultrafiltration-HPLC method

目的对灯盏花素脂质体进行质量评价，测定灯盏花素脂质体包封率。方法采用超滤法分离脂质体与游离药物；采用Kromasil ODS柱（200mm×4．6mm，5μm），流动相为甲醇-乙腈-20mmol·L^-1磷酸盐缓冲液（pH值为2．5）（体积比为17：17：66），流速为0．8mL·min^-1，检测波长为334nm，测定药物含量，计算包封率。结果超滤法能很好的将脂质体与游离药物分离，游离药物的平均回收率在95．9％～97．6％，加样回收率在96．4％～97．1％，脂质体不能透过超滤膜；该色谱条件下，灯盏乙素得到良好分离，辅料不干扰测定，灯盏乙素在1．0～40．0mg·L^-1内线性关系良好（r=0．9999），日内和日间RSD均小于2．0％（n=5），加样回收率在99．7％～100．1％之间，RSD小于1．23％。结论该方法可用于灯盏花素脂质体的质量控制。

Objective To evaluate the quality of liposomal formulation of breviscapine and develop the method for the determination of entrapment efficiency. Methods Ultrafiltration was applied to separate the free breviscapine from the liposomes; The content of scutellarin was determined by HPLC with Kromasil C18 column. The mobile phase was methanol-acetonitrile-20 mmol·L^- 1 phosphate buffer（pH2.5） （ V： V ： V = 17 ： 17：66）with the flow rate of 0.8 mL·min^-1. The UV detective wavelength was set at 334 nm. Results The free breviscapine was well separated from the liposomes with stirred ultrafiltration cell, the breviscapine recoveries of ultrafiltration were between 95.9 % -97.6 %, and the recoveries with blank liposome were between 96.4 %-97.1%. The liposomes could not permeate the ultrafiltration film. The HPLC method had good linearity in the range of 1.0- 40.0 mg·L^-1（ r = 0. 999 9）. Both the intra-day RSD and inter-day RSD were less than 2.0 % （n = 5）, the recoveries of scutellarin with blank liposomes were between 99.7 % - 100.1%, the RSD were less than 1.23 %. Conclusions The method can be used for the quality control of liposomal formulation of breviscapine.