农产品中三种氰基菊酯类农药残留的化学发光酶免疫分析方法研究

摘要

Synthetic pyrethroids are widely used in domestic, public health, agricultural, forestry, and veterinary applications for their remarkably high insecticidal activity and low mammalian toxicity compared with organochlorinated and organophosphate compounds.The α-cyano pyrethroids such as deltamethrin, cypermethrin, λ-cyhalothrin, fenpropathrin and fenvalerate,are quickly becoming the dominant.Although pyrethroids are thought to be safe for humans,due to the extensive human use and release of synthetic pyrethroids, their toxic effects on ecological, environmental, and human health have become increasingly concerned.Therefore,it is necessary to determine α-cyano pyrethroid residues in the environment and agricultural products for regulatory and monitoring purposes.

目录

声明

Abstract

Table of Contents

Chapter 1 General introduction

1．1 OVERVIEW

1．2 DEVELOPMENT OF ENZYME LINKED IMMUNOASSAY

1．2．1 Hapten design and synthesis

1．2．2 Antigen preparation

1．2．3 Production of monoclonal antibody

1．2．4 Immunoassay format

1．2．5 Competitve ELISA dose-response curve

1．2．6 Data analysis

1．2．7 Optimization of assay conditions

1．2．8 Application to real sample analysis

1．3 DEVELOPMENT OF CHEMILUMINESCENCE ENZYME IMMUNOASSAY

1．3．1 Overview

1．3．2 General principles

1．3．3 Chemiluminescence immunoassay

1．4 α-CYANO PYRETHROID PESTICIDES

1．4．1 Overview

1．4．2 Toxicity of pyrethroid pesticides

1．4．3 Analytical methods of pyrethroid pesticides

1．5 STUDY OBJECTIVE

Chapter 2 2 Preparation of a broad-specific monoclonal antibody for three Q-cyano pyrethroids and characterization by enzyme-linked immunosorbent assay

ABSTRACT

2．1 INTRODUCTION

2．2 MATERIAL AND METHODS

2．2．1 Reagents and Chemicals

2．2．2 Buffers and Solutions

2．2．3 Instruments

2．2．4 Animals

2．2．5 Synthesis of hapten

2．2．6 Preparation of immunogens and coating antigen

2．2．7 Production of monoclonal antibodies

2．2．8 Indirect ELISAs

2．2．9 Influence of physicochemicai factors on immunoassays

2．2．10 Cross-reactivity

2．3 RESULT AND DISCUSIONS

2．3．1 Hapten synthesis and mAb production

2．3．2 Optimization of the ELISA

2．3．3 Assay sensitivity and specificity

2．4 CONCLUSION

Chapter 3 3 Development of chemiluminescent enzyme immunoassay for rapid detection of three α-cyano pyrethroids

ABSTRACT

3．1 INTRODUCTION

3．2 MATERIAL AND METHODS

3．2．1 Reagents and chemicals

3．2．2 Buffers and solutions

3．2．3 Immunoassay procedure

3．2．4 Optimization of chemiluminescent enhanced solution

3．2．5 Assay optimization

3．2．6 Cross reactivity

3．3 RESULTS AND DISCUSSION

3．3．1 Chemiluminescent enhanced solution (CES)

3．3．2 CLEIA optimization

3．3．3 Assay Sensitivity and Selectivity

3．4 CONCLUSION

Chapter 4 4 Evaluation of the CLEIA for rapid detection of α-cyano pyrethroid residue in agricultural samples

Abstract

4．1 INTRODUCTION

4．2 MATERIAL AND METHODS

4．2．1 Reagents and chemicals

4．2．2 Sample pre-treatment for ELISA and CLEIA

4．2．3 Sample pretreatment for GC analysis

4．2．4 GC-ECD conditions

4．3 RESULTS AND DISCUSSION

4．3．1 Matrix interferences

4．3．2 Analysis of spiked samples

4．3．3 Comparison of CLEIA and ELISA

4．4 CONCLUSION

Chapter 5 5 General conclusions and future perspectives

5．1 SUMMARY

5．2 GENERAL DISCUSSION AND PERSPECTIVES

5．2．1 Assay performances of CLEIA

5．2．2 Assay application to real samples

5．2．3 Future perspectives for CLEIA

5．2．4 Future perspectives for broad-specific immunoassays

REFERENCE

Acknowledgements

Curriculum Vitae