摘要
为筛选肺靶向多西他赛脂质体（DTX-LP）的包封率测定方法,本研究结合固体分散与泡腾技术制备DTX-LP,马尔文粒度仪测定其粒径分布和Zeta电位;采用高效液相色谱法测定DTX含量;考察DTX在不同媒介中的溶解情况;对比高速离心法和鱼精蛋白凝聚法测定DTX-LP的包封率。研究表明,制得的DTX-LP平均粒径为（0.72±0.23）μm,Zeta电位为-27.5 m V,表面荷负电。DTX在0.5～100μg/m L浓度范围内与峰面积呈良好的线性关系（r=0.999 9）,日内精密度和日间精密度均〈2.00%,室温放置0～8 h内稳定性RSD≤2.00%,回收率在99.96%～108.05%之间。当以生理盐水、0.2%吐温80的生理盐水、0.5%吐温80的生理盐水作为媒介,通过高速离心法测得DTX-LP的包封率分别为（91.15±1.07）%、（85.28±3.75）%、（79.76±2.71）%（n=3）,而鱼精蛋白凝聚法测得DTX-LP的包封率分别为（95.46±0.29）%、（93.61±2.75）%、（87.83±1.23）%（n=3）,生理盐水能够完全溶解微量的、未包封的游离药物。通过分析对比,选择鱼精蛋白凝聚法、以生理盐水作媒介,更适用于亚微米级、表面荷负电的DTX-LP包封率的测定,能避免表面活性剂及高速离心对脂质体膜结构的影响,该方法操作简单、快速、可行性好、准确度高。
To choose a method for the determination of encapsulation efficiency of lung-targeted docetaxel liposomes（DTX-LP）,DTX-LP was prepared by solid dispersion method combining with effervescent technology,and its size distribution and Zeta charge were determined by Malvern Zeta-sizer.HPLC method was established to measure the content of DTX.The solubility of DTX in different solutions was tested.High speed centrifugation method and protamine-induced aggregation method were used to determine the encapsulation efficiency of DTX-LP.The research showed that the average size of obtained DTX-LP was（0.72±0.23） μm while Zeta potential was-27.5 m V with negatively charged surface.A good linear relationship was found between the peak area and the concentration of DTX ranging from 0.5 μg/m L to 100 μg/m L（r=0.999 9）.Intra-day and inter-day precision RSD were both less than 2.00%.The stability RSD stored in ambient temperature within 8 h was no more than 2.00%.The recovery rate of DTX was in a range of 99.96%～108.05%.Using normal saline,0.2% tween 80-normal saline,0.5% tween 80-normal saline as solvents,the encapsulation efficienc y of DTX-LP determined by high speed centrifugation method were（91.15 ±1.07）%,（85.28 ±3.75）% and（79.76 ±2.71）%（n =3）,respectively;while those determined by protamine-induced aggregation method were（95.46±0.29）%,（93.61±2.75）% and（87.83±1.23）%（n=3）,respectively.A small amount of non-encapsulated drugs could be completely dissolved by normal saline.Through analyzing and comparing,protamine-induced aggregation method might be more applicable to determine the encapsulation efficiency of negatively charged DTX-LP with normal saline as the solvent.It could avoid the influence of surfactant and high centrifugation to the membrane structure of DTX-LP.The proposed method could be simple,rapid,feasible and with high accuracy.
作者
刘忠洪
张梨
王丽娟
余瑜
Liu Zhonghong;Zhang Li;Wang Lijuan;Yu Yu(College of Pharmacy, Chongqing Medical University, Chongqing, 400016)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2018年第5期2276-2281,共6页
Genomics and Applied Biology
基金
国家自然科学基金(No.81172097)资助
关键词
多西他赛
脂质体
包封率
鱼精蛋白凝聚法
高速离心法
Docetaxel
Liposome
Encapsulation efficiency
Protamine aggregation method
High speed centrifugation method
作者简介
通讯作者:余瑜,yuyu3519@163.com
