Studies On The Methods Of AChE Immobilization And Its Application On Rapid Detection Of Organophosphorus And Carbamate Pesticides

Organophosphorus and carbamate pesticides are two common pesticides which are still widely used in agriculture production. They can inhibit AChE activity in body and cause poisoning symptom, even endanger the life of human being. So, pesticide residues in food have payed more attention over the world. Some virulent kinds of organophosphorus and carbamate pesticides are still used to vegetables and fruits in some regions although it forbidden by the government, the toxicosis incidents of human and animal resulted from taking food which contains high level of pesticide residues are happened usually.Although GC and HPLC have high veracity and sensitivity for detecting pesticides residues, but they can't adapt to rapid detection in market and field scene for its expensive instruments, professional performance, rigorous operating environment and long time consuming. Traditional rapid detection method have some shortcomings such as the enzyme(AchE) difficult to storage and the color of extracting solution can affect the detection, which cause much inconvenience to apply. So research a rapid, sensitive and convenient method for detection pesticide residues is our imperative needs, it will play an important role in prevention high pesticide residues farm products from the marketplace and to reduce the incidents of food safty.The aim of this study is to search a convenient, rapid, sensitive, credible and practical method for detection the pesticide residues in fruits and vegetables at markets, which can adapt to the produce and sale status of agriculture product in our country. The results are as follow:(1) The method of activity determination of AchE was optimized, and the activity determined parameters of dissociative enzyme and immobilized enzyme were improved. The AchE activity determined system are as follows: 50μL enzyme solution (one immobilized enzyme card), 50μL zymolyte AschI, 50μL visualization reagent DTNB, 2.9mL pH8.0 phosphate buffer solutiori(PBS) (3.95mL for immobilized enzyme detection system), and 1%SDS termination reagent 1mL(none for immobilized enzyme detection system).(2) Nitrocellulose membrane (NCM) was selected from the 5 materials to immobilize...