Extraction method for acetylcholinesterase and application thereof

Abstract

An extraction method and applications of acetylcholinesterase relate to a method to extract acetylcholinesterase from carp and a method to detect the residue of organic phosphorous pesticides. The extraction method is that: the tissues of carp are undergone homogenate by phosphorous acid buffer solution in ice bath, and is froze and centrifuging, and then the enzyme liquid is prepared. The detecting method is that: acetylcholinesterase of carp brain is used for an enzyme source, 2, 6-dichloro Indophenol Sodium salt is used for a color developing agent, qualitative filter paper is used for a vector, and the enzyme is immobilized by absorption. The detailed operation is that: a plastized hard piece of paper is stuck with small pieces of paper; one side of the hard paper is added with the color developing agent and acetylthiocholine iodide, the other side acetylcholinesterase solution, and the hard paper is put into a frigirater to be froze and dried and can be fetched when needed; the eluant of organic phosphorous pesticide is dripped on and fully wet two filter papers, and the two pieces of filter paper are overlapped for minutes to observe if the color is changed; the color of the filter paper changes from blue to colorless if organic pesticide is not existed, otherwise the blue color does not fade or does not obviously fade, the higher the concentration is, the more obviously the color change is. The invention has the advantages of low cost, rapid detecting speed and high sensibility.

Description

technical field [0001] The invention relates to a method for extracting acetylcholinesterase from the brain, muscle and liver of crucian carp and a method for rapidly detecting residual organophosphorus pesticides by using it. Background technique [0002] Existing "A kind of preparation method for extracting acetylcholinesterase from animal blood" (patent application number 02134464), comprises the following process steps: the blood of rat, chicken, duck, cow, dog, rabbit, eel, eel Solidify at room temperature, freeze, take the supernatant, and centrifuge the supernatant to obtain the crude extract of acetylcholinesterase, purify it by ammonium sulfate precipitation, use dialysis to remove ammonium sulfate, concentrate and dry the enzyme solution, and obtain purified acetylcholine Esterase products. [0003] Acetylcholinesterase is used to detect pesticide residues, such as "A Recombinant Acetylcholinesterase and Its Preparation Method and Detection Method" (Patent Applica...

AI Technical Summary

Problems solved by technology

There is also acetylcholinesterase as the enzyme source, 2,6-dichloroindophenol acetate as the color reagent, and 2,6-dichloroindophenol acetate as the color reagent, which is not yet available in China, and the price is expensive. Up to 1,000 yuan or more

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