分子内与分子间协同作用型红绿光双发射ECL染料标记物研究

Intermolecular interaction electrochemiluminescence (ECL) for the Ru(bpy)32+ and propylamine (TPA) system has been studied extensively and found broad applications in commercial technology. Unfortunately,TPA is usually needed in vast molar excess (100 mM) in the system due to its continuously consumption during the oxidation process, and this high excess TPA employed will inevitably cause some increased background noise. To solve this problem, an intramolecular activating reductant, phenothiazine, which is known to have a reversible one electron oxidation potential, and a tertiary nitrogen atom similar to that of TPA in its structure, is utilized to replace TPA to covalently link with novel bimetallic ruthenium-iridium complexes, which are designed by connecting normal red emission ruthenium and green emission iridium ECL labels through a flexible saturated carbon chain,to get completely redox reversible, red/green dual emission ECL labels. In the proposal, both concerted intra- and intermolecular ECL performance and ratio ECL detection method will be investigated and utilized to significantly decrease the amount of TPA coreactant needed, overcoming the disadvantage of the commercial technology such as intermolecular interaction ECL alone and single detecting wavelength etc. A concerted intra- and intermolecular ECL with dual emission ratio ECL detection method, novel bimetallic ruthenium-iridium ECL labels with independent intellectual property rights will be built up and employed for pratical detection to check its applicability. All these can provide a general methodology to further improve and tune the ECL efficiency by using of the bimetallic complexes linked by flexible saturated carbon chain, lead to some fundamental understandings for dual emission and ratio ECL detection method.

常规电化学发光(ECL)体系利用三联吡啶钌与还原剂三丙胺(TPA)的分子间作用，TPA氧化的不可逆导致其在检测中不断消耗，必须大量添加，并且TPA会自氧化产生背景干扰。本申请将红、绿光发射的联吡啶钌、铱染料连成一体，并引入吩噻嗪（有与TPA类似的叔胺结构，氧化还原可逆）作为分子内还原剂，设计氧化还原可逆型红绿光双发射ECL染料标记物。借助标记物自身分子内、标记物与外加TPA的分子间协同ECL，最大限度降低体系中TPA的添加量；并利用红绿光ECL比例消除TPA自氧化的背景干扰，提高检测灵敏度，彻底解决常规ECL体系单纯分子间作用、TPA大量添加、单波长检测的弊端。最后对新型标记物进行应用测试，考察其效果。该项目将获得具有自主知识产权、分子内与分子间协同作用型、双发射比例ECL染料标记物，构建分子内与分子间协同作用下的双发射比例ECL检测方法，填补比例ECL检测空白，丰富ECL检测机理。

常规电化学发光(ECL)体系利用三联吡啶钌与还原剂三丙胺(TPA)的分子间作用，TPA 氧化的不可逆导致其在检测中不断消耗，必须大量添加，并且 TPA 会自氧化产生背.景干扰。本申请将红、绿光发射的联吡啶钌、铱染料连成一体，并引入吩噻嗪（有与 TPA 类似的叔胺结构，氧化还原可逆）作为分子内还原剂，设计氧化还原可逆型红绿光双发射 ECL染料标记物。借助标记物自身分子内、标记物与外加 TPA 的分子间协同 ECL，最大限度降低体系中 TPA 的添加量； 并利用红绿光 ECL 比例消除 TPA 自氧化的背景干扰， 提高检测灵敏度，彻底解决常规 ECL 体系单纯分子间作用、TPA 大量添加、单波长检测的弊端。最后对新型标记物进行应用测试，考察其效果。该项目将获得具有自主知识产权、分子内与分子间协同作用型、双发射比例 ECL 染料标记物，构建分子内与分子间协同作用下的双发射比例 ECL 检测方法，填补比例 ECL 检测空白，丰富 ECL 检测机理。开发了核酸、溶媒体等亚细胞器探针，以及部分金属离子、氧化剂、还原剂荧光检测方法。发表SCI论文19篇，获授权中国发明专利4件。