Studies On Analysis Of Red Tide Toxion By Capillary Electrophoresis-Enzyme Immunoassay

This thesis studied on the analysis of Red Tide toxins, with the conjunction of Capillary Electrophoresis(CE) and Enzyme Immunoassay. Neurotoxic Shellfish Poisoning(NSP) and Ciguatera Fish Poisoning(CFP) were analyzed, based on the reaction between o-aminophenol(OAP) and H2O2 catalyzed by Horseradish Per- oxidase(HRP). Then the mixture of four kinds of shellfish toxins was separated and deteced with the introduction of AuNPs as labels. The four kinds of shellfish toxins were Paralytic Shellfish Poisoning(PSP), Diarrhetic Shellfish Poisoning(DSP), Amnesic Shellfish Poisoning(ASP) and NSP.The whole thesis can be divided into seven chapters:Chapter 1 The introduction of the basic theories of CE, the recent advances in the study of Enzyme Immunoassay and the present research of the Red Tide toxins.Chapter 2 The studies on the improvement of the Electrochemical(EC) detection electrode, the technology of the manufacture of acidized Poly(vinyl alcohol) coated capillary and the combination of the Pt electrode.Chapter 3 The studies on the OAP-H2O2-HRP system with EC detection used in Electrophoresis-Enzyme Immunoassay(CEIA). In this chapter the optimum experiment conditions such as concentration and pH of buffers, separation voltage(Vs), detection Electrical Potential(Ed) and injection conditions were studied. On the optimum conditions, the the linear range of free HRP was 2.3×10^-12-3.5×10-10 mol/L, the detection limit is 1.09×10^-12 mol/L and the detection limit of quality can even reach the level of zmol. Chapter 4 The studies on a new methord for detection of NSP by CEIA, using OAP-H2O2-HRP reaction system. By the specific recognation between antigen(Ag) and antibody(Ab), the detection of Ag* and Ab-Ag* conjuction was achieved. Then the competitive immune reaction mode was used in the qualitative and quantitative detection of NSP in the scallop sample. By this methord, the linear range of NSP was 1.0-50.0 ng/mL, and the detection limit was 0.1 ng/mL which was 4 times lower than ELISA.Chapter 5 The studies on a new methord for the detection of CFP and the fish sample by CEIA based on the amplification effect of gold nanoparticles(AuNPs). Ab* was achieved by immobilizing the enzyme(HRP) and Ab to the surface of AuNPs which was used as solid carrier. Because of the high specific surface area, large amount of HRP was immobilized, and the sensitivity of detection was also improved. By the specific recognation between Ag and Ab, the detection of Ab* and Ag-Ab* conjuction was achieved. Then the non-competitive immune reaction mode was used in the qualitative and quantitative detection of CFP in the fish sample. By this methord, the linear range of CFP was 1.0-50.0 ng/mL, and the detection limit was 0.3 ng/mL.Chapter 6 The studies on a new methord for the separation of the 4 kind of shellfish toxins by CEIA. As labels AuNPs influenced the migration rate of the components, which can improve the separation efficiency of CE. Then by the competition between DSP, PSP, NSP toxin in shellfish sample and their Ag* to combind with corresponding Ab and ASP toxin combinding with its Ab*, the separation and detection of the toxins were achieved.Chapter 7 was the conclusion of the whole work.