注射用多西他赛纳米粒质量控制技术研究

发布时间：2018-09-11 19:54

【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。
[Abstract]:Objective: as a new drug delivery carrier, nanoparticles have the ability to improve the absorption of unstable or insoluble drugs, prolong the internal circulation of drugs, and increase the ability of drugs to pass through the biofilm barrier. Increasing drug targeting has become a hot spot in the research of anti-tumor drug carriers. However, particle leakage and degradation products may occur in preparation, transportation and storage of nanoparticles. At present, there is still no standard and uniform quality standard for nanoparticles. In this paper, the quality control technology of doxetacemide granules for injection is studied, and the physicochemical properties of nanoparticles (including appearance, particle size distribution and Zeta potential determination, acid value, acid value) of nanoparticles are studied. Peroxide value), content determination, entrapment efficiency, in vitro release, residual solvent, harmful impurity lysophosphatidylcholine were systematically investigated, which provided reference for quality control of nanoparticles. Methods: (1) the physicochemical properties of doxetacernet for injection were investigated, the appearance of nanoparticles was examined, the particle size distribution was measured by static light scattering and dynamic laser scattering, and the Zeta potential of nanoparticles was measured. The acid value and peroxide value of nanoparticles were investigated. (2) HPLC method was used to determine the content of doxetacinamil for injection and the methodology was studied. (3) the entrapment efficiency of nanoparticles was determined and the method of separating free drugs from nanoparticles was studied. Free drugs and nanoparticles were separated by ultracentrifugation, microcolumn centrifugation, dextran gel column chromatography and dynamic dialysis, respectively. The effects of various methods on the determination of entrapment efficiency of doxetacernet for injection were studied. (4) in vitro release assay of doxetacernet for injection; With reference to Chinese Pharmacopoeia 2010 Edition, part II, appendix X C, the third method was used to determine the dissolution rate of a hemodialysis bag containing doxetacemide for injection and to fix it on a small dissolving paddle. According to the cumulative release amount, the release rule of doxetacemide for injection in vitro was investigated. (5) Headspace gas chromatography was used to determine the residual solvent in nanoparticles. (6) the content of lysophosphatidylcholine, a harmful impurity in doxetacemide granules for injection, was determined by HPLC detector. Results: the appearance of doxetacemide injection was white and the color was uniform, the average diameter of the nanoparticles was 117.9028 nm by dynamic laser scattering, and the particle size larger than 200nm was less than 5% Zeta potential (-3.54 MV), and the average particle size was 117.9028 nm by dynamic laser scattering (DLS). The acid value of nanoparticles was not more than 2, peroxide value was less than 3, the content of doxetacemide for injection was determined by HPLC method, and the entrapment efficiency was 95.0 by dynamic dialysation-high performance liquid chromatography. The kinetic models for in vitro release of doxetacemide for injection were well fitted, and the residual solvent ethanol content in nanoparticles was determined by headspace gas chromatography with 0.2% ethanol content. The content of lysophosphatidylcholine in nanoparticles was determined by HPLC method. The content of lysophosphatidylcholine in nanoparticles was not detected by HPLC evaporative light detector. Conclusion: the established method is simple to determine the physical and chemical properties of doxetasine granules for injection. The rapid HPLC method is used to determine the content of nanoparticles, the dynamic dialysation-HPLC method is used to determine the encapsulation efficiency of nanoparticles, and the dissolution cup method is used to determine the release of nanoparticles in vitro. The method of headspace gas chromatography for the determination of residual solvent HPLC-UV and HPLC-ELSD method for the determination of lysophosphatidylcholine in nanoparticles is feasible, reliable and controllable. It can provide a reference for the establishment of quality standard for doxetasine granules for injection.
【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943

【参考文献】