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The pyrethroid pesticide immune and chromatographic analysis method

Author: HuLiMing
Tutor: YangYaLing
School: Kunming University of Science and Technology
Course: Chemical Engineering
Keywords: Pyrethroid enzyme-linked immunosorbent assay (ELISA) Solid Phase Extraction (SPE) HPLC
CLC: TQ450.2
Type: Master's thesis
Year: 2009
Downloads: 45
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Abstract


Stable, free from unusual odor, safe, and low in concentration, pyrethroid is a type of bionic pesticide which is widely used in agriculture, industry as well as in the prevention and control of furniture pests. However, more and more animal experiments indicate that pyrethroid may impair neural development, induce cancer, suppress immunity system and interfere incretion system. Therefore, the determination of pyrethroid residue in crops, food and environment is of special significance. With pyrethroid as the analyte, this thesis studies the immunity analysis method and chemical and instrumental analysis methods of this pesticide.This paper gives an overview of the progresses achieved in studying the determination and analysis methods of pyrethroid pesticide, including the pre-treatment of samples, testing and analysis of chemical instrument, and immunity analysis. It then summarizes the properties and characteristics of immunity analysis and its application in the testing of biological, pharmaceutical, foodstuff, and environmental samples.In the part of experiment, deltamethrin hapten and antigen were synthesized by adopting the method recommended by Sally K. Mak et al. The experimental animal was immunized by antigen and a monoclonal antibody (McAb) specific for deltamethrin was produced by hybridoma technology. Then the chapter goes on to investigate the specificity of McAb by ELISA and SDS-PAGE. The results showed that PRD-McAb was of the 1gG1 type; that its molecular weight was 168 Kda.determined by SDS-PAGE; that the affinity constant of McAb with coated complete antigen was 4.75×108 L/mol; and that the antibody is differential.In the base of getting the McAb, we determined and analyzed the PRD by ELISA. After optimizing reaction condition, we obtained a calibration curve for free PRD by ELISA. Then, with Tie guan yin Tea bought from the supermarket as the sample, we detected the recoveries of PRD. The optimum condition of ELISA was determined at 37℃in temperature,1.5 hours in length, and pH7.6 in solution. The minimum detection limit was found to be 89 pg/mL by ELISA method and the recovery rate was above 90%. Furthermore, the biotin-avidin amplifying system was applied to establish a more sensitive determination method, the minimum detection limit of which was 5 pg/mL.We uesd’SPE in the pre-treatment of the tea sample, and established PRD-SPE-HPLC off-line analysis system which was then used in testing four kinds of pyrethroids. We optimized the condition of analysis in PRD-SPE-HPLC system, and chose methanol as the elution solvent, and 70% Acetonitrile/water as the analytical solvent. When tested by PRD-SPE-HPLC system, the recovery rate of four PRD pesticides in tea was above 80% while that of one pesticide isomer was 72.9%. Then this chapter makes a comparative study of the PRD-SPE-HPLC system and ELISA method in terms of sample treatment and sensitivity. We found that immunoassay method is high sensitive, simple and fast in determinating pyrethroid pesticide. Chapter Five summarizes the research process and anticipates what needs to be done in future researches. Combination of biotechnology and chemical methods will be a trend of analyse.

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CLC: > Industrial Technology > Chemical Industry > Pesticide Industry > General issues > Determination of biological activity and safety evaluation
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