A Hybrid Electrochemiluminescence System And A New Method For Oxygen Fluorescence Analysis

Electrogeneranted chemiluminescence ?ECL? analysis method has the advantages of good controllability, high sensitivity, and the instruments is simple and cheap. Therefore, widely used in biological sensing analysis. Currently reported most of the electrogeneranted luminescence biosensors are based on a single electrochemical luminescence material, which is unfavorable to set up an electrochemical luminescence biological sensing method to detect multi-component, also limits the development of the electrochemical luminescence biological sensing analysis. At present, the electro-chemical luminescence system mainly include luminol-H₂O₂ and Ru?bpy?₃²⁺-TPrA system in biological sensor application, people are also studying new electrochemical luminescence material, such as metal iridium complexes. In order to research a new electrochemical luminescence system, this project is "Hybrid electrochemical luminescence system and oxygen fluorescence analysis research of new methods" entitled to carry out the related research work. This topic completed with supported in the national natural science fund "Microfluidic bipolar electrode electrochemical luminescence bio-sensor new method research" ?No.21275095? and "Electrochemical luminescence biosensor some basic problems of research" ?No.21475082?.This project consists of introduction and research of two parts, the first part is the first chapter, for the introduction part of the project; The second part for the second and the third chapter, chapter of the paper research part and the fourth chapter is the conclusion of this project.The first chapter is the introduction part, mainly on the electrochemical luminescence analysis methods and common luminescence mechanism and the electrochemical luminescence system has carried on the brief overview; Common electrochemical luminescence system are reviewed in this paper the research progress of related applications; This paper briefly introduces the common technology of oxygen sensor and detection method, the fluorescence quenching of the oxygen sensor detection principle and the common characteristics of the fluorescence quenching of oxygen sensing materials. Fluorescence quenching is reviewed in this paper some applications of oxygen sensor in the biology and the research progress of oxygen sensors in recent years; Finally, introduces the purpose, significance and research content of the project.The second chapter "The electrochemical luminescence behavior of hybrid electrochemical luminescence system research", in the traditional luminol-H₂O₂ and Ru?bpy?₃²⁺-TPrA luminescence system research foundation, through a certain proportion mixed, study the electrochemical luminescence behavior of the hybrid system. The experimental condition is optimized choice; The experimental results show that when the concentration of the TPrA is 35 mM and the concentration of H₂O₂ is 1mM, can detect Luminol and Ru?bpy?₃²⁺?with good potential of material of two kinds of signal resolution and realize the ECL detection at the same time. Research has found a new ECL system:Luminol-TPrA system. And the ECL behavior of the system to do the research, research has shown that low concentration of Luminol has greatly enhance function of TPrA ECL; In the presence of high concentration of Luminol, the ECL intensity of luminol enhance, ECL of TPrA is weakened; TPrA can weakly promote the electrochemical luminescence of Luminol. The system can be used in ammonia class material testing, has a certain practicality. The research work on the new method for the analysis of double potential distinguish electrochemical luminescence research provides some reference data.The third chapter "Metal iridium complexes ?pq?₂Ir?psa? oxygen analysis research of new methods", this new laboratory experiments found that oxygen have an effect on the metal iridium complexes ?pq?₂Ir?psa?. The fluorescence intensity has strong quenching effect. Based on this phenomena, to study the ?pq?₂Ir?psa? light of the physical properties, in acetonitrile and water solution of O₂ response. The experimental results show that both in acetonitrile and water solution, the O₂ make the fluorescence ?pq?₂Ir?psa? quenched, the quenching constants in acetonitrile K_SV= 52.47, can be used in the detection of oxygen. Through the analysis of ultraviolet spectrum, the quenching belongs to a dynamic quenching. Also studied the light fastness of the ?pq?₂Ir?psa?, the reversibility of dissolved oxygen quenching. Finally, using the ?pq?₂Ir?psa? to dyeing of HeLa cell, and evaluate the oxygen content in HeLa. The experimental results show that the ?pq?₂Ir?psa? can cross the HeLa cell membranes, and give the cytoplasm staining, in the green excitation ?520?550 nm? wavelength, the fluorescence is orange red. The research work laid the foundation for the reagent electrochemical luminescence research and provides some new ideas for certain oxygen sensing method research.