| With the incidence rate of malignant tumor is higher and higher, the malignant tumor is one of the difficult problems in current human must face, so now antitumor drug R & D in the medical field occupies an important position. In recent years, Chinese medicine in the field of anti-tumor has some progress, in the chemical structure, traditional Chinese medicine has certain diversity, and mechanism of different traditional Chinese medicine on tumor is different, so, traditional Chinese medicine in antitumor activity has great potential. Oxymatrine is from the legume roots of Sophora flavescens Ait or flat, wide plant roots are obtained by extraction and separation of alkaloids and Oxymatrine has the characteristics of high rate of low toxicity, has a very effective anti swelling Tumor effect. Although the evident in the anti-tumor effects, but there are to after the medicine biological utilization degree is low, and the required dose and other shortcomings, in the widely used has some limitations,therefore, we use the layers of self assembling method of oxymatrine packaging prepare Oxymatrine capsule, thereby increasing drug bioavailability, reduce the dosage.Part one:Objective: to establish a method for the determination ofoxymatrine layer by layer self assembly of microcapsules encapsulation efficiency high performance liquid chromatography, and determination of the entrapment efficiency. Method: chromatographic column: thermo Sycronis C18, flow phase: Acetonitrile-0.2% phosphoric acid solution with three amine to adjust p H to 80(13:87,V/ V), detection wavelength:220nm, flow phase velocity: 1ml/min. The low temperature, high speed centrifugation separation of the free drug microcapsules. Results:oxymatrine in 20-300ug/mL range, the linear relationship between the benign Good. The average recovery rate is 99.63%,, the precision RSD≤1.74%. Conclusion: The high performance liquid chromatography method was established to meet the requirements of HPLC, using high performance liquid chromatography can accurate determination of LBL method for the preparation of oxymatrine content of oxymatrine in microcapsules and microcapsules encapsulation rate, which is simple, fast, high specificity,sensitivity and strong advantages.The second part:Objective: the layer by layer self characterization of microcapsules prepared by self-assembly method and physicochemical properties of preliminary inquiry. Methods: using projection electron microscopy of layer by layer self loading microcapsules were observed,and determination of the particle diameter. The potential instrument of each layer of the assembly of surface potential were measured. Examine the body of the drug in different time drug cumulative release amount, andexplores the effect of assembling layers of oxymatrine drug release.Results: through the projection electron microscopy of LBL microcapsules,showing by LBL assembled microcapsules particle size and dispersion,satisfied uniformity to micro capsule, further by Zeta potential instrument of assembly The each layer of the surface potential were measured. The results show that with opposite charge of polyelectrolyte successfully assembled on the surface of the template core, then we measured the LBL loaded microcapsules 3-180 min time period of drug average cumulative release amount, results show, OM drugs from microcapsules release and no burst release, but from microcapsules release slowly, releasing effect.Further we compare the assembly 2,5,7,10 layer loaded microcapsules in3-180 min time drug average cumulative release amount, assembly 2 layer of microcapsules in 60 min drug release completely average cumulative release amount of 72. 4±2.35(%), assembly 5 layer of microcapsules in100 min basic complete release average cumulative release amount of 75.1±0.26(%), assembled 7 layer of microcapsules in 100 min basic complete release average cumulative release amount of 74.5±0.26(%), assembled 10 layers of drug loaded microcapsules in 180 basic complete release average cumulative release amount of 1.03±76.4(%), shows that the more layers of packaging, and the surfaces of the capsule poly electrolyte membrane more thick, the sustained release effect is more obvious.The third part:Objective: to layer upon layer self-organizing loadedmicrocapsules cells and drug concentration were measured to explore its effect on cancer cells. Methods: using high performance liquid phase chromatography established intracellular drug concentration standard curve,determination of drug loaded microcapsules and free micro capsule on cancer cells after intracellular concentration changes and comparison of oxymatrine. Results:Intracellular drug concentration in the liquid in good0.625-10ug/mL regional linear relationship, correlation coefficient is0.99879, and the average recovery rate was 98.2%, stability samples RSD≤2.14% and inter day RSD ≤ 1.8%, the intra day precision RSD≤2.1%,the drug concentration in extracellular fluid in good 0.1-5.0ug/mL regional linear relationship, related coefficient is 0.99975, and the average recovery rate of 94.5%, stability samples RSD is less than or equal to 1.64%, inter day RSD ≤2.8%, intra day precision(RSD) was less than or equal to 2.5%,the HPLC method,specific, high accuracy. The comparison results show that the layer by layer self assembly method of oxymatrine capsule can be faster and more effective in MCF-7 cells, which may and into LBL film material on MCF-7 cells with specific relevant. |
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