脂质体中蛋白类药物包封率的测定Research on the detection of encapsulation efficiency in protein-loaded liposomes
李可欣,陈大为,商捷,赵秀丽,庞大海
LI Ke-xin1,2,CHEN Da-wei2,SHANG Jie1,ZHAO Xiu-li2
摘要(Abstract)：
目的建立脂质体中蛋白类药物包封率的测定方法。方法以牛血清白蛋白(bovine serum al-bumin,BSA)为模型蛋白,采用逆向蒸发法制备了平均粒径为5.129μm的BSA脂质体。采用离心-参照法测定BSA脂质体的包封率,并以凝胶过滤、超速离心、超滤和透析4种常用方法对包封率的结果进行比较。结果该方法平均回收率为101.7%,RSD为2.32%(n=5),BSA质量浓度在1.02～1 507.5 mg.L-1内线性关系良好。离心-参照法测定3批BSA脂质体的平均包封率为69.74%,RSD为1.34%。而其他几种方法测定同一批脂质体的包封率值偏高或偏低,即测定方法存在问题。结论该法简便灵敏,测得值真实、可靠,可以用于蛋白类药物脂质体中包封率的测定。
Objective To establish the method for the determination of encapsulation efficiency(EE%)in protein-loaded liposomes.Methods The liposomes loading bovine serum albumin(BSA) was prepared as model protein using the modified reverse-phase evaporation method.The mean particle-size was 5.129 μm.The centrifuge-reference method was used for the first time in which empty liposomes were as the reference to eliminate the ingredient′s influence.Additionally,gel filtration,ultracentrifugation,ultrafiltration and dialysis were used to compare the drug EE%.Results The mean recovery was 101.7%,RSD was 2.32%(n=5).The linear range was good in 1.02-1 507.5 mg·L-1.The mean EE% was 69.74%,RSD was 1.34%.But other four methods were lower or higher than the true value.Conclusions The new established method is suitable and accurate,which can be developed for the determination of the drug EE% of the protein-loaded liposomes.
关键词(KeyWords)：
脂质体;牛血清白蛋白;包封率;考马斯亮蓝
liposome;bovine serum albumin;encapsulation efficiency;Bradford
基金项目(Foundation):
作者(Author):
李可欣,陈大为,商捷,赵秀丽,庞大海
LI Ke-xin1,2,CHEN Da-wei2,SHANG Jie1,ZHAO Xiu-li2
DOI: 10.14066/j.cnki.cn21-1349/r.2010.09.015
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