柠檬苦素白蛋白纳米粒制备工艺优化及其分子表征

Optimization of preparation process and molecular characterization of limonin-albumin nanoparticles using response surface methodology

[目的]构建柠檬苦素白蛋白纳米粒,为进一步应用柠檬苦素提供理论依据。[方法]以牛血清白蛋白为纳米载体,通过去溶剂化—化学交联法制备柠檬苦素白蛋白纳米粒,以包埋率为指标,通过单因素试验和响应面试验确定最佳制备工艺,并对纳米粒进行表征。[结果]当牛血清白蛋白添加量为64 mg,柠檬苦素添加量为4 mg,醇水体积比为5∶1,交联剂添加量为125μL时,交联固化后得到的柠檬苦素白蛋白纳米粒平均粒径为(152.2±0.5)nm,多分散指数为0.196±0.026,包埋率为(79.21±0.36)%;通过低压透射电子显微镜观察到的柠檬苦素白蛋白纳米粒呈现圆球形,而傅里叶红外光谱及X射线衍射结果表明柠檬苦素以无定型或者无序状态被成功包裹在白蛋白中;该柠檬苦素白蛋白纳米粒悬浮液具有良好的贮藏稳定性。[结论]采用去溶剂化—化学交联法制备的柠檬苦素白蛋白纳米粒贮藏稳定性良好。

[Objective]The construction of limonin-albumin nanoparticles provides a theoretical basis for the further application of limonin.[Methods]Limonin-albumin nanoparticles were prepared by desolvation-chemical crosslinking method using bovine serum albumin as nanocarriers.The optimal preparation process was determined by single factor experiment and response surface test with embedding rate as index,and the nanoparticles were characterized.[Results]When the amount of bovine serum albumin was 64 mg,the amount of lemon bitter was 4 mg,the specific volume ratio of alcohol to water was 5∶1,and the amount of crosslinker was 125μL,the average particle size and polydispersion index of limonin nanoparticles were(152.2±0.5)nm and 0.196±0.026 respectively.The embedding efficiency was(79.21±0.36)%.The limonin-albumin nanoparticles observed under low-pressure transmission electron microscopy showed a spherical shape.Infrared spectroscopy and X-ray diffraction results showed that limonin was successfully encapsulated in albumin in an amorphous or disordered state.The limonin-albumin nanoparticle suspension has good storage stability.[Conclusion]The limonin-albumin nanoparticles prepared by desolvation-chemical crosslinking method had good storage stability.