【作者】 谢玲玲；

【导师】 罗卓雅；

【作者基本信息】 广州中医药大学 ， 药物分析学， 2014， 硕士

【摘要】 目的：纳米粒作为新型药物给药载体,具有改善口服不稳定或难溶性药物的吸收、延长药物体内循环时间、增加药物穿过生物膜屏障的能力,增加药物靶向性等优点,成为研究抗肿瘤药物载体的热点。而纳米粒制剂在生产制备、运输、贮存都可能发生粒子泄漏、害降解产物产生等问题。目前对纳米粒制剂仍无规范统一的质量标准,本文对注射用多西他赛纳米粒制剂质量控制技术进行研究,对纳米粒理化性质(包括纳米粒外观、粒度分布及Zeta电势测定、酸值、过氧化值)、含量测定、包封率、体外释放、残留溶剂、有害杂质溶血磷脂酰胆碱检测方法进行系统考察,为纳米粒制剂质量控制提供参考依据。方法：(1)对注射用多西他赛纳米粒理化性质进行考察；检查纳米粒外观；采用静态光散射法和动态激光散射法测定纳米粒粒度分布,测定纳米粒Zeta电势；考察纳米粒酸值及过氧化值。(2) HPLC法测定注射用多西他赛纳米粒含量,并进行方法学研究；(3)测定纳米粒包封率,对游离药物与纳米粒分离方法进行研究；分别采用超速离心法、微柱离心法、葡聚糖凝胶柱层析法及动态透析法分离游离药物及纳米粒,并研究各方法对测定注射用多西他赛纳米粒包封率结果的影响。(4)注射用多西他赛纳米粒体外释放测定；参照《中国药典》2010版第二部附录X C溶出度测定第三法将装有注射用多西他赛纳米粒混悬液置透析袋内并固定在溶出小桨上,根据累积释放量探寻注射用多西他赛纳米粒体外释放缓释规律。(5)顶空气相色谱法测定纳米粒中残留溶剂；(6)采用HPLC不同检测器测定注射用多西他赛纳米粒中有害杂质溶血磷脂酰胆碱含量。结果：检查注射液多西他赛纳米粒外观为白色冻干块状物,色泽均一；采用动态激光散射法测得纳米粒平均粒径为117.9028nm,大于200nm的粒子不超过5%,Zeta电势为-3.54mV；纳米粒酸值结果不大于2,过氧化值不大于3; HPLC法测定注射用多西他赛纳米粒含量为9.94mg· g-1,动态透析-高效液相色谱法测定纳米粒包封率为95.0%；初步考察注射用多西他赛纳米粒体外释放各级动力学模型拟合较好；顶空气相色谱法测定纳米粒中残留溶剂乙醇含量为0.2%；超声、离心提取磷脂组分, HPLC法紫外测定纳米粒中溶血磷脂酰胆碱含量为0.24%, HPLC蒸发光检测器未能检出纳米粒中溶血磷脂酰胆碱含量。结论：建立的注射用多西他赛纳米粒制剂理化性质检查方法简单、快速；HPLC法测定纳米粒含量、动态透析-HPLC法测定纳米粒包封率、溶出小杯法测定纳米粒体外释放、顶空气相色谱法测定纳米粒残留溶剂、HPLC-UV及HPLC-ELSD法测定纳米粒中有害杂质溶血磷脂酰胆碱含量方法可行、可靠、可控,可为注射用多西他赛纳米粒制剂建立质量标准提供参考依据。更多还原

【Abstract】 Objective:Nanoparticles as a novel drug delivery vehicle. They have so many advantages, such as, improved absorption of oral unstable drugs and poorly soluble drugs, prolonged drug circulation time in vivo, increased the capacity of the drugs through the biof ilm barrier, increased drug targeted and so on. that are widely used in the field of anti-tumor. At production and preparation,transportion and storage of nanoparticles, leakage may occur, which could produce harmful degradation products. Currently, there is no uniform quality standards for the nanoparticles preparation. In this parper, studied of quality control technology of nanoparticles of docetaxel for injection, nanoparticles’physical and chemical properties (including nanoparticles appearance, size distribution and zeta potential, acid value, peroxide value), assay, encapsulation efficiency, in vitro release, residual solvents, harmful impurities lysophosphatidylcholine detection, to provide reference quality control for nanoparticles preparation.Methods(1)Studied of physicochemical properties of nanoparticles of docetaxel for injection;Evaluation of the appearance of nanoparticles;Determination of particle size distribution and zeta potential of nanoparticles used dynamic light scattering instrument and Static light scattering method;study of acid value and peroxide value of nanoparticles;(2)Used HPLC method to assay of nanoparticles of docetaxel for injection, and established methodology validation;(3) Studied the separation methods of free drug and nanoparticles to evalue nanoparticles’encapsulation efficiency; Studied of ultracentrifugation, microcolumn centrifugation, sephadex column chromatography and dynamic dialysis method to separate free drug and nanoparticles, choosed the most suitable method for determining entrapment efficiency of nanoparticles of docetaxel for injection.(4) The determination of the condition release in vitro of nanoparticles of docetaxel for injectionAccording to the third mehod of dissolution in the second2010edition of "Chinese Pharmacopoeia", dialysis bag with suspension of nanoparticles was fixed on the stripping small oar, put it in the mediumto sudy the release codition of nanoparticles.(5) Determination of the residual ethanol by different injection methods of GC.(6) Determination of the content of lysophosphatidylcholine with two different types detector of high performance liquid chromatography,ultraviolet detector and evaporative light scattering detector.ResultsThe appearance of nanoparticles of docetaxel for injection was white freeze-dried lumps and color uniformity;The average particle size of nanoparticles of docetaxel for injection was about117nm, and particle size that more than200nmwas less than5%, Zeta potential was about-3.54mv measured by dynamic laser scattering method;Acid value not more than2and peroxide value not more than3;The assay of nanoparticles of docetaxel for injection was9.94mg·g-1using the HPLC method;Preliminary study on vitro release of nanoparticles, and according to cumulative release in vitro release was fitted for order all kinetics equation;The contect of residual ethanol in nanoparticle preparation was0.2%by the headspace gas chromatography method, that accordance with ICH;Extractiing phospholipid composition by ultrasound and centrifugal and then detected the contect of lysophosphatidylcholine, that was0.24%by HPLC-ELSD method, while it wasn’ t detected using HPLC-UV method.ConclusionThe established methods of nanoparticles of docetaxel for injection, such as the physicochemical properties, assay, encapsulation efficiency determination using dynamic dialysis-HPLC, the condition release in vitro, the detection content of harmful impurities of lysophosphatidylcholine is feasible, reliable and controllable, that could provide basis for the quality standard reference of nanoparticle preparation.更多还原