壳聚糖—海藻酸盐纳米粒子制备及其对BSA包载与释放研究.docVIP

摘要 本论文利用带相反电荷的聚电解质在水溶液中通过静电相互作用自 组装形成壳聚糖－海藻酸盐纳米粒。利用动态光散射纳米粒度分析仪考 察了钙离子及壳聚糖对粒子粒径的影响。结果显示钙离子的存在可使粒 子粒径从（268.5±15.5）nm 降为（203.4±14.8）nm，但随着钙离子含量 的继续升高，粒径迅速增大。当钙离子浓度大于 0.45g/L 时形成凝胶。 壳聚糖含量的增加和蛋白的包裹均会使粒径增大。所制备的纳米粒对 BSA 具有较高的包载能力，并有一定的缓释作用。当壳聚糖投料量增加 时，可使 BSA 在 pH＝7.4 的 PBS 中的释放减慢。 文中还考查壳聚糖的存在对紫外吸收 A280、Lowry、Bradford 三种 蛋白定量检测方法的影响。用三种方法检测含有不同浓度、分子量、脱 乙酰度的壳聚糖的 BSA 溶液，将测定结果与理论值进行比较。结果显 示，当待测蛋白溶液中含有壳聚糖时，三种方法的测定结果都高于实际 蛋白含量，其中 A280法受影响最大。壳聚糖对 A280及 Lowry 法的干扰与 其结构中乙酰胺的含量有关，提高脱乙酰度可使其对紫外吸收和 Lowry 两种方法的干扰减弱，但对 Bradford 法无显著影响。A280 和 Lowry 法可 通调整对照液消除干扰影响。 关键词：壳聚糖，海藻酸钠，纳米粒子，聚电解质复合物，蛋白定量检 测 ABSTRACT Nanoparticles were prepared by dropping alginate solution into CaCl2 solution and then coated by chitosan. A dynamic light scattering particle size analyzer was used to study the size of the nanoparticles prepared with different relative mass ratios of sodium alginate, calcium chloride and chitosan. The mean diameter of the particles prepared without calcium chloride was（268.5±15.5）nm. The particle size decreased to (203.4±14.8）nm when a low concentration of calcium chloride was added. However, a calcium chloride to sodium alginate mass ratio less than 0.3 was necessary to form nanoparticles rather than hydrogel. The mean diameter of the particles increased with the concentration of chitosan . The nanoparticles with more chitosan had a bigger size and exhibited a slower release of BSA. Also，chitosan (CS) was tested for interference with the absorbance at 280nm (A280 method), Lowry’s and Bradford’s methods of protein determination. When CS existed in the bovine serum albumin (BSA) solution, the determining results based on A280 were much higher than the real values and the bias increased with the concentration of CS. CS with lower deacetylation degree (DD) and larger molecular weight (Mw) had a more significant interference effect. CS had a similar influence on the Lowry’s method but the bias was much less compared to those of A280 method. There was a constant positive bias in the re