核酸适体修饰的亲水超高交联整体柱电色谱高效分析痕量双酚A的研究

For improving the efficiency and specificity of molecular recognition on the organic polymerized monolith, the researchs on novel aptamer-modified hydrophilic hypercrosslinked monolith and new model for the on-line specific enrichment and separation with affinity capillary electrochromatography (CEC) would be done by using bisphenol A (BPA) as the representative object. By means of the surface modification of aptamer on the skeleton of hydrophilic hypercrosslinked monolith, the microstructures and characteristics of the monolith will changed and a desired functional monolith will be perfect. Numbers of active sites for highly specific affinity chromatography are expected, and specific identification of BPA on this hydrophilic hypercrosslinked monolith will be also obtained. With the comprehensive application of EOF and auxiliary pressure for the continuous sample injection, the analytic technique and separation behavior of CEC with the functional monolith will be investigated. A novel on-line micro-analysis method is investigated, and several significant advantages including the enrichment of electric field-amplified action, adsorption of functional stationary phase, and the high-efficient separation of affinity CEC, will be obtained. The problems existing in the traditional enrichment mode of organic polymerized monolith, such as small column capacity, large sample consumption and zone broadening, will be well avoid. It is favorable for the highly specific recognition, efficient enrichment and rapid separation of trace BPA. Besides, by using the sensitive laser-induced fluorescence (LIF) detection method, CEC-LIF hyphenated technology would be established. It is highly sensitive and competent for the trace analysis of BPA. It might be promoted for enrichment and affinity separation of BPA in environmental, and probes a new approach for high-efficient analyses of trace contaminant with aptamer affinity electrochromatography. Finally, satisfactory result including 5-8 research paper and 2-3 patents will be expected.

以双酚A为研究对象，从功能材料和分析模式着手，创新研发新型核酸适体修饰的亲水超高交联整体柱及其亲和电色谱在线特异富集、分离的新模式，致力于新型有机聚合整体柱高效富集、特异识别的研究。项目首次将核酸适体亲和作用引入亲水超高交联整体柱，在高相比的超高交联聚合结构表面修饰双酚A的核酸适体，形成大量的核酸适体高特异性亲和作用位点，大幅提高整体柱亲和识别特性，研发出特异识别的高相比亲和整体柱；联用电色谱高效分析技术，建立集电场堆积富集、高相比固定相吸附和亲和电色谱分离为一体的新型电色谱在线微分析模式，解决常规聚合整体柱分子识别作用相比较小、富集效率较低、样品富集区带展宽明显等瓶颈问题，建立痕量双酚A的高特异性识别和高效分离技术。在此基础上，集成激光诱导荧光柱上检测模式，实现环境中痕量双酚A的超高交联整体柱核酸亲和电色谱高灵敏特异亲和分析。拟发表高水平论文5-8篇、发明专利2-3项。

以双酚A为研究对象，项目从“功能柱制备”-“作用机制研究”-“特异识别分析”着手，围绕“多作用模式分离介质创制及作用机制探究”和“核酸适配体键合整体柱研发及特异识别分析”，开展了亲水多模式新型聚合整体柱、核酸适配体键合整体柱及其特异识别技术的创新研究，建立了亲水高交联核酸适体键合整体柱制备技术和双酚A（BPA）特异识别技术，相关成果在J. Chromatogr. A等发表SCI论文8篇，获得国家发明专利授权2项，培养硕士3名，较好地完成了研究目标。.1. 开展了多作用模式功能材料的创制及作用机制探究。针对常规聚合整体柱普遍疏水性较大、作用基团结构较为单一，极性酚类物质分离柱效较低的不足，创新研制了以离子单体为功能基团的高亲水有机聚合整体柱、纳米笼状硅胶倍半体超高交联聚合整体柱及动态改性整体柱等功能材料5项，探讨了整体柱聚合结构、分离性能和作用机制，构建并实现了酚类在功能整体柱上的色谱保留、氢键作用、电泳迁移和离子选择多重作用模式，为强极性环境下双酚A的高效分离提供了高效的分离介质及理论支撑。.2．开展了核酸适配体键合整体柱研制及分离模式构建。基于环氧开环反应，或纳米金键合技术，柱后修饰5’-CCGGTGGGTGGTCAGGTGGGATAGCGTTCCGC GTATGGCCCAGCGCATCACGGGTTCGCACCA-3-NH2适配体；基于点击化学反应，原位键合5’-SHCC GGTGGGTGGTCAGGTGGGATAGCGTTCCGCGTATGGCCCAG CGCATCACGGGTTCGCACCA-3’。创新研制了系列核酸适配体键合POSS基高交联聚合整体柱，研发了稳定高效的核酸适体键合整体柱3项，实现了双酚A适配体的高效键合和聚合整体柱的特异识别功能化。.3. 开展了核酸适配体整体柱对双酚A（BPA）的识别分析研究，结合应用荧光柱上检测模式，实现了强极性环境下核酸配体键合整体柱的高灵敏特异亲和分析，痕量BPA激素得到了特异性分离和检测，突破了传统有机共价聚合整体柱相比小、特异识别作用弱的不足与限制，为环境中痕量双酚A的核酸适体整体柱高效分析研究提供了相关的科学探索和技术支撑。