微柱离心法测定新型口服抗脑瘤前药CAT3油酸缀合物固体脂质纳米粒的包封率  

Determination of Encapsulation Efficiency of Novel Solid Lipid Nanoparticles Containing Oleic Acid-CAT3 Conjugates by Mini-Column Centrifugation Method

作　　者：王洪亮 徐雪晴 王汝冰 高丽丽 刘玉玲 WANG Hong-liang;XU Xue-qing;WANG Ru-bing;GAO Li-li;LIU Yu-ling(State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院北京协和医学院药物研究所天然药物活性物质与功能国家重点实验室,药物传输技术及新型制剂北京市重点实验室,北京100050

出　　处：《中国药学杂志》2020年第12期1021-1026,共6页Chinese Pharmaceutical Journal

基　　金：十三五国家科技重大专项资助(2018ZX09711001-002-005);北京协和医学院青年创新基金资助(2017350020);中国医学科学院医学与健康科技创新工程项目资助(CIFMS-2019-I2M-1-005)。

摘　　要：目的建立一种微柱离心的方法测定新型口服抗脑瘤前药CAT3油酸缀合物固体脂质纳米粒(OA-CAT3-SLN)与普通CAT3固体脂质纳米粒(CAT3-SLN)的包封率。方法采用葡聚糖凝胶(Sephadex G-50)微柱离心法分离固体脂质纳米粒(SLN)中包封药物与游离药物,通过洗脱曲线确定包封药物与游离药物分界点确定洗脱方式与洗脱液用量:以1 mL水,共洗3次将SLN包封药物完全洗脱,再以2 mL乙醇,共洗3次将游离药物完全洗脱。洗脱下来的CAT3以HPLC-UV进行测定,并计算包封率。本实验以洗脱方法的回收率与重复性为指标进行了方法学验证,并对OA-CAT3-SLN与CAT3-SLN各3批进行了包封率测定。结果微柱离心法能有效实现固体脂质纳米粒中包封药物与游离药物的分离。方法回收率为(99.64±1.97)%(n=9);包封率重复测定结果为(83.71±0.70)%(n=5)。OA-CAT3-SLN与CAT3-SLN包封率分别为(86.26±2.65)%与(72.22±4.52)%(n=3)。结论建立的分离方法操作简便,回收率高,重复性好,对不同制备工艺具有区分性,测定结果可靠。OBJECTIVE To establish a mini-column centrifugation method to determine the encapsulation efficiency(EE)of novel solid lipid nanoparticles containing oleic acid-CAT3 conjugates(OA-CAT3-SLN)and normal CAT3 SLN(CAT3-SLN).METHODS Sephadex G-50 mini-columns were used to separate the encapsulated drug and free drug in the solid lipid nanoparticles(SLN)with the help of centrifugation.The boundary point of the elution between the encapsulated drug and free drug was established by the elution curve.The encapsulated drug in the SLN was eluted with 1 mL water for three times.Then 2 mL of ethanol was used to elute the separated free drug for three times.The eluted CAT3 was quantified by the verified HPLC-UV method and used to calculate the EE.The method was verified with the recovery and repeatability test.Finally,the EE of three batches of OA-CAT3-SLN and CAT3-SLN were determined.RESULTS The mini-column centrifugation method could effectively separate the free drug from the encapsulated drug in SLN.The column recovery was(99.64±1.97)%(n=9),and the result of EE repeatability test of OA-CAT3-SLN was(83.71±0.70)%(n=5).The EE of OA-CAT3-SLN and CAT3-SLN were(86.26±2.65)%and(72.22±4.52)%,respectively(n=3).CONCLUSION The established separation method is simple and reliable,with high recovery and good repeatability,and can distinguish different preparation processes.

关 键 词：菲并吲哚里西啶 前药 油酸 缀合物 固体脂质纳米粒 包封率 微柱离心法 

分 类 号：R944[医药卫生—药剂学]