Application Of Chemiluminescence In Pharmaceutical And Biological Analysis

Chemiluminescence assay has been successfully applied in different fields of chemical analysisbecause of its simple instruments, automation and other characteristation. The combination of chemiluminescence assay with other technologyhas attracted reachers'attention and extensively promote its development. In this thesis, we employed chemiluminescence for pharmic analysis and established an universal chemiluminescent analytical method for acidic drug in basic environment. Furthermore, we proposed an ultrasensitive chemiluminescent immunoassay based on the combination of chemiluminescence and nanotechnology. The proposed methods and the constructed sensors exhibit broad application prospect in biology and pharmacy. The research sections were as following.1. IntrodutionIn this section, the basical knowledge, application in pharmaceutical of chemiluminescence, and the development of the combination of nanotechnology and chemiluminescence were summarized. First, the development, conception, principle and basic installation of chemiluminescence were introduced detailedly. Furthermore, we have described the applications of chemiluminescence in pharmaceutical analysis, and the applications of nanomaterials in chemiluminescence field. At last, the works and innovations of this dissertation were presented.2. Determination of metoclopramide by flow-injection chemluminescence methodThe metoclopramide could only be dissolved in acid solution, so the quantificational detection of metoclopramide by chemiluminescent assay in basic environment is difficult. Potassium dichromate can oxide metoclopramide, and the Cr(Ⅵ) could be reduced to Cr(Ⅲ), which could be act as the enhacer for luminol-H2O2 system. Therefore we established a method for the determination of metoclopramide by the combining with folw injection. The results showed that the method exhibited good stabilization and wide linear range of 1.0×10-8-1.5×10-5 g/mL with a detection limit of 8.0×10-9 g/mL. This method has been successfully used to the determination of metoclopramide in drug, demonstrating high sensitivity and good rapidity of analysis.3. Determination of human immunoglobulin G by a rapid and ultrasensitive chemiluminescent immunoassay by the passive mixing and gold nanoparticles-colloidal carbon nanosphere hybrid materials labelsA novel, sensitive and rapid flow injection chemiluminescent immunassay has been developed based on the gold nanoparticles-colloidal carbon nanosphere hybrid material (Au NPs@C) as platforms for the immobilization of second antibody and horseradish oxidase, and the paramagnetic microbeads (PMs) as platforms for the immobilization of first antibody. The fluidic system combined with a three-dimensional helical glass tube and two spiral glass tubes was benefit for the rapid analysis of the present immunassay. The prepared Au NPs@C hybrid material was expected to offer a promising template for increasing the number of immobilized biomolecule because of its satisfactory chemical stability and good biocompatibility. The detection could be completed within 10min due to the fluidic system combined with a three-dimensional helical glass tube for rapid mixing of immunoreagents and two spiral glass tubes for magnetic separation. The free horseradish peroxidaselabeled composite was separated from sandwich immunocomplex formed on paramagnetic particles by a permanent magnet and then mixed with chemiluminescent substrate in a long spiral tube, and then the chemiluminescent signal. Resulted from the detection mixture was collected. Employing Human immunoglobulin G(HIgG) as an analyte, the immunoassay could be completed within 10 min with a linear calibration range of 1.0-1000 ng/mL with a detection limit of 0.74 ng/mL. The developed assay method was versatile, rapid, offered enhanced performances, which could be easily extended to other protein detection as well as DNA analysis.4. Synthesis, characterization and chemiluminescence application of GO/luminol nanocompositesA new method for the preparation of luminol-functionalized graphene has been proposed, and a novel chemiluminescent sensor was constructed in the presence of chitosan. Luminol was adsorbed on the surface of graphene, and the luminol-functionalized graphene/chotison film could react with hydrogenperoxide to generate chemiluminescence. In this experiment, we compared the three methods for the preparation of graphene/luminol. It was found that the chemiluminescent intensity of luminol-functionalized graphene which was prepared in the present of EDC and NHS at room temperature exhibited the strongest chemiluminescent intensity. As the reagent for forming film, chitosan was better than sol-gel to prepare luminol/graphene/chotison film. Finally, the film was immolilized on the inner surface of glass column to constructed a chemiluminescent sensor for the determination of hydrogenperoxide in the range of 5.0×10-6-2.0×10-4 mol/L with the detection limit of 2.7×10-6 mol/L. Due to the graphene's unique properties, abundant chemical bonds and the excellent biocompatibility of the chitosan, the sensor demonstrate great potential in the applications of biosensors.