| Chemiluminescence(CL) detection,based on the CL reaction of the analyte,was applied widely in biology,medicine,and environment for its' high efficiency and sensitivity.The dynamic character is a special point of CL,that is,the photons were emitted going with the process of reaction.So,in analytical applications,while CL was used as the method of quantitive assay,the reproducibility tends to be a problem for the difficult control of the reaction.As a solution treatment method in non-equilibrium state,the Flow Injection Analysis(FIA) could be coupled with CL, overcoming the poor reproducibility of CL and making the assays automatically. Another point,which could not be neglected,is the interference of the coexisted reagents in a special CL reaction system.It makes an unideal selectivity for traditional CL analysis.Chemiluminescence coupled with Capillary Electrophoresis(CE),a new and high performance separation method,would make the analysis selective and sensitive.In this dissertation,we coupled the chemiluminescence with flow injection and capillary electrophoresis to research the application of CL in DNA and protein analysis.The optimized analytical conditions and operation steps had been gained,as a valuable reference for the further application of CL in biological analysis.This dissertation is composed of four chapters as following:Chapter 1:Introduced the main principles of chemiluminescence and the Common CL reaction systems;the applications of chemiluminescence in the assay of DNA,protein,food and medicine;the coupling methods of CL with Flow-Injection, and the high performance separation technologies.In the end,pointed out the purpose and meaning of this dissertation.Chapter 2:Magnetic separation based Flow Injection-Chemiluminescent DNA analysis with copper sulfide labels.Based on the super separation and enrichment ability of magnetic beads(MBs), we proposed a new Flow Injection Chemiluminescence(FI-CL)analytical system for DNA detection by using CuS nanoparticles as tags.The assay relies on the hybridization of target DNA with probel(CuS-ssDNA conjugate)and probe2 (MB-ssDNA conjugate)to form a sandwich structure.The hybridization events were then monitored by the CL intensity catalyzed by the resolved Cu2+ions from these CuS nanoparticles anchored on sandwich hybrids.Under the optimum conditions,the linear response range was 1.0×10-11～1.6×10-9mol/L for the perfectly complementary target ssDNA with detection limit(LOD) of 3×10-12mol/L.The standard deviation (RSD) for 1.0×10-9 mol/L complementary ssDNA was 3.2%(n=11,).Such proposed FI-CL DNA biosensor could recognize the target DNA sequences quite well.Chapter 3:Building and realization of a CE-CL analytical system.As a new,high performance and low-cost separation method,Capillary Electrophoresis has been used widely in biology,medicine,environment,and food safety assays.The nano-liter of the analytical samples makes the detection method difficult and challenging.CL analysis is characterized by ultrasensitivity,simplicity and low-cost,due to no exiting light sources,avoiding the effects of stray light and instability of light source,and uniquely suited for high sensitive detection of CE. Based on the IFFL-D FI-CL analytical system and high voltage equipment in our laboratory,we build up a useful CE-CL analytical system by hand.This system worked well,and the detection data was gained with satisfactory.Chapter 4:The detection of Hemoglobin(human) based on the CE-CL system.Based on the CE-CL detection system,built with our hand as described in the previous chapter,the detection of hemoglobin was done as a sample.The detection result was accurate with high-reproducibility,indicated that this CE-CL system was stable and reliable.The flow-path and the pH of the CE-CL assay were chosen.In 22 minutes,the detection could be finished.Linear rang:1×10-8～1×10-5 mol/L with the limit of detection(S/N=3):3×10-9 mol/L.Practical samples of blood and serum were detected with discussion of the results.
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