| In the review of this thesis, the basic principles, characteristics and the applications of chemiluminescence are briefly introduced. The flow injection (FI-CL) is becoming more and more popular owing to its simple instrumentation, high sensitivity, wide dynamic range, reproducibility, simplicity and rapidity. New flow injection chemiluminescence methods coupled with other techniques such as immobilized enzyme, immunoassay, micro reaction cell, renewable surface assay were built for the determination of ascorbic acid, serum cholesterol and Immunoglobulin G based on luminol and hydrogen peroxide CL systems. This thesis chose horseradish peroxidase(HRP) and hemoglobin(Hb) as typical enzyme of peroxidase and mimetic peroxidase. The research section includes three subsection as following:1. A chemiluminescence suppression method for the determination of ascorbic acid in Vc tablet based on Luminol-HRP-H2O2-ascorbic acid system with immobilized enzyme was developed. The linear range for ascorbic acid is 4.0×10-8～4.0×10-4mol/L and the detection limit is 1.6×10-8mol/L. The sampling frequency was 50 sample/h. The relative standard deviation (n=10) is 2.4 % for 4.0×10-7mol/mL ascorbic acid. A satisfactory result could be obtained using the novel method for the determination of ascorbic acid in Vc tablet.2. Hemoglobin was used as a mimetic peroxidase in the catalytic chemiluminescence reaction of lumino and hydrogen peroxide. The hemoglobin-catalyzed reaction was coupled with the oxidation reaction of cholesterol catalyzed by COA(cholesterol oxydase, COA.) to determine total cholesterol (TC) and free cholesterol (FC) in human serum by flow injection chemiluminescence method. The sampling frequency was 30 s/h. The linear range for free cholesterol is 5.6×10-7mol/L～3.8×10-5mol/L and the detection limit is 3.0×10-7mol/L. The linear range for total cholesterol is 1.1×10-6mol/L～4.3×10-5mol/L and the detection limit is 1.5×10-7mol/L.3. A sequential injection renewable surface heterogeneous enhance chemiluminescence immunoassay system with flow-through reaction column was developed for the determination of human immunoglobulin G(IgG) in serum. Immobilized antibody was prepared by conjugation of sheep anti-human IgG antibody (the first antibody) to protein A coated sepharose 4B beads. Horseradish peroxidase labeled sheep anti-human IgG antibody was used as the second antibody. Two antibody reacted with human IgG antigen and formed a sandwiched antibody-antigen conjugate which in the flow-through reaction column. IgG in human serum was detected according to the chemiluminescence system of luminol-HRP- H2O2 which is based on a catalytic effect of HRP bounded in the conjugate. After a measurement, the beads (45-165μm) were discharged from the column, and the column was ready for the next determination after flushing with buffer. Under the optimized conditions, a detection limit of 0.01mg/L IgG was achieved. The linear dynamic range of calibration curve is in the 0.05～3.0 mg/L IgG. The method was applied successfully to determine IgG in human serum samples.
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