【作者】 肖尚友；

【导师】 夏之宁；

【作者基本信息】 重庆大学 ， 生物医学工程， 2006， 博士

【摘要】 相互作用在化学体系、生物化学体系和环境体系中广泛存在,是各种生理活动得以正常进行和生理功能得以正常发挥的基础。毛细管电泳具有分离效率高、分析速度快、样品用量少等特点,在研究分子相互作用方面具有独到的优势。因此开展相互作用研究,特别是毛细管电泳相互作用分析及应用研究在手性药理学、环境毒理学、药物创制学及生物医学工程领域有重要意义。相互作用有多种表达方式,包括结合常数、结合位点及自由能变等参数,其中结合常数是表征相互作用强弱最重要的参数之一。本文在以毛细管电泳为手段研究分子相互作用的基础上,发展了分子相互作用的毛细管电泳分析(CEIA)方法,特别是针对复杂体系和电中性分子的相互作用分析方法。将CEIA方法应用于小分子与生物大分子、小分子与细菌和鸡血红细胞的相互作用分析,以获得相互作用结合常数。论文进一步基于相互作用的原理,开展了以可溶性淀粉、糊精和麦芽糖为手性添加剂的西酞普兰毛细管电泳手性分离,并对手性识别相互作用机理进行了较为深入的研究。具体主要内容包括:(1)应用毛细管电泳方法,使用十二烷基硫酸钠胶束赋予中性客体分子对-硝基苯胺和2-氨基吡啶有效淌度,研究了中性主体分子18-冠醚-6与对-硝基苯胺和2-氨基吡啶的相互作用,建立了中性分子相互作用的毛细管电泳分析方法。以毛细管电泳峰漂移方法,测得了中性主体与中性客体分子之间的相互作用结合常数。(2)将毛细管电泳相互作用分析方法从一种物质与另一种物质的相互作用的测定发展到多个组分共存时与另一物质的相互作用测定,达到了同时评价各组分与指定分子的结合能力的目的。通过这种方法测定了大黄中的有效成分大黄素、芦荟大黄素和大黄酚与β-环糊精和羟丙基-β-环糊精的结合常数。(3)应用毛细管电泳区带灌注-自标记技术,研究了麻黄碱、咖啡因、对乙酰氨基酚与牛血清白蛋白的相互作用以及麻黄碱与鸡血红细胞的相互作用。(4)通过对细菌样品预处理方法、缓冲液种类、检测波长以及聚合物添加剂对分离度影响等的考察,研究了大肠杆菌与金黄色葡萄球菌的毛细管电泳行为,获得了优化的毛细管电泳分离条件,实现了大肠杆菌与金黄色葡萄球菌的快速分离,从而建立起一种辅助的细菌定性识别方法。进一步,研究了抗菌药物阿奇霉素与金黄色葡萄球菌的相互作用,发现阿奇霉素能与金黄色葡萄球菌的表面蛋白发生相互作用,从而引起金黄色葡萄球菌淌度的变化。(5)基于竞争相互作用原理,采用非手性的C18色谱柱,通过在流动相中加入手性选择剂β-环糊精的方法实现了西布曲明对映体的拆分。西布曲明对映体在更多还原

【Abstract】 Molecular interactions exist widely in chemical, biochemical and environmental systems. Bioactivities and biological functions are based on these molecular interactions. Capillary electrophoresis has unique advantages of efficient separation, fast speed and small amount of samples required, in studying molecular interactions. Therefore, studies and applications of molecular interactions by capillary electrophoresis are of significant importance for chiral pharmacology, environmental toxicology, drug discovery and biomedical engineering.The interactions can be characterized by many parameters such as binding constants, binding sites and free energy etc. Hoewer, the binding constants is one of the most important parameters to characterize the interactions.In this thesis, an approach called capillary electrophoresis interaction analysis (CEIA), especially for interactions among those neutral-electronic molecules in complex systems, was established. Binding constants of small molecules with biomacromolecules, with bacteria and red blood cell are determined by CEIA. Based on the principle of molecular interactions, chiral separation of citalopram hydrobromide was successfully achieved,by using amylose,dextrin and maltose as chiral additives;The mechanism of chiral recognition by maltose was studied further. The main contents of this thesis are listed below:(1) According to the different mobilities caused by sodium dodecylsulfonate, interactions of neutral host molecule 18-crown-6 with p-nitroaniline and 2-aminopyridine were studied by CEIA respectively. The CEIA method of neutral molecules was established. The binding constants of host molecules with guest molecules were determined by capillary electrophoresis peak-shifting method.(2) The CEIA method was expanded to determine molecular interactions of components in complex systems,which was able to evaluate binding abilities of an interested component with all other components within one test. The binding constants of bioactive components from Rhubarb such as Emodin, Aloe-emodin and Chrysophanol withβ–cyclodextrin and hydroxypropyl-β-cyclodextrin were determined by this method.(3) The partial-filling-self-marking technique (PFSM) was employed for the interaction analysis of ephedrine, caffeine and acetaminophen with Bovine Serum更多还原