花椒麻味物质的分子印迹分离纯化机理研究与应用

Chinese prickly ash and its relating products were widely used in the field of food industry, mainly due to its numb taste. However, quantitative detection methods of numb-taste substance were limited in application and research directly on account of no commercial availability of standard product both at home and abroad. This project is to develop a systematical purification technology of numb-taste substance in Chinese prickly ash using molecular imprinting method. Based on the previous studies, the reference substance and its corresponding substitute were employed as the template molecule which was separated by column chromatography and liquid chromatography etc. The computer technology was applied in identifying the configuration with minimum energy by the method of Semi-expirical-PM3 to optimize configuration of template molecule, functional monomer and the complex. Molecularly imprinted polymer was acquired by bulk polymerization based on the matrix of nanometer materials. Subsequently, the polymer was treated with structure and performance characterization, and then with actual separation. Verifications of separated substance were conducted by HPLC and GC-MS etc, and the performance of established molecular imprinting methods were evaluated and improved in order to resolve hard problems of less binding sites in a "tadpoles" type and molecularly imprinting purification with big stereo specific blockade. Therefore, these research achievements, helpful for the establishment of Chinese prickly ash quality testing system, were of great value to the development and application of molecular imprinting technique.

在各个领域使用花椒及花椒加工制品，主要是取其香麻味，因为国内外还无花椒麻味物质标准品销售，限制了其定量检测方法的研究与推广运用。本项目是在课题组以前研究基础上，以柱层析法、液相色谱法等技术分离得到的花椒麻味物质对照品和获得的替代品为模板分子；采用计算机技术，应用Semi-expirical-PM3方法优化模板分子、功能单体及其复合物构型，筛选出能量最小的构型；以纳米材料为基质，采用本体聚合等方法制备分子印迹聚合物；再对该分子印迹聚合物进行结构和性能表征及实际分离；利用HPLC、GC-MS等对分离物质进行验证；对建立的分子印迹方法的分离效果进行优化和改进并完善；等等，从而建立花椒麻味物质的分子印迹纯化技术，解决具有"蝌蚪"型的少结合位点、空间位阻大的分子结构特点物质的分子印迹纯化难题。因此，本项目研究成果，有利于建立花椒质量检测体系，并对分子印迹技术的发展和运用具有重要的理论价值。

在各个领域使用花椒及花椒加工制品，主要是取其香麻味，因为国内外还无花椒麻味物质标准品销售，限制了其定量检测方法的研究与推广运用。本项目是在课题组以前研究基础上，以柱层析法、液相色谱法等技术分离得到的花椒麻味物质对照品和获得的替代品为模板分子；采用计算机技术，应用Semi-expirical-PM3方法优化模板分子、功能单体及其复合物构型，筛选出能量最小的构型；以纳米材料为基质，采用本体聚合等方法制备分子印迹聚合物；再对该分子印迹聚合物进行结构和性能表征及实际分离；利用HPLC、GC-MS等.对分离物质进行验证；对建立的分子印迹方法的分离效果进行优化和改进并完善；等等，从而建立花椒麻味物质的分子印迹纯化技术，解决具有"蝌蚪"型的少结合位点、空间位阻大的分子结构特点物质的分子印迹纯化难题。现在所有研究内容已经完成，取得了如下研究成果：（1）建立了花椒麻味物质分子印迹分离方法和中试生产线，已获得花椒麻味物质标准品3250克，供应并满足了全国18家相关企业的需要；（2）获得了基质材料、模板分子、功能单体及其聚合物构型之间的优化模型，从而建立具有“蝌蚪”型结合位点少且分布不均的分子结构特点物质的分子印迹分离纯化的实验研究方法体系；（3）已公开发表了学术论文9篇，其中SCI收录论文4篇，EI收录论文4篇，申请了国家发明专利1项，培养博士研究生2名和硕士研究生2名，获得教育部2017年度高等学校科学研究优秀成果奖（科学技术）——技术发明一等奖1项（已于2017年12月公示）。因此，本项目全面完成了项目任务书的要求和指标，并且本项目研究成果，有利于建立花椒质量检测体系，并对分子印迹技术的发展和运用具有重要的理论价值。