Immobilization acetylcholinesterase tablet and matched substrate tablet, preparation method and use for the same

Abstract

The present invention provides a immobilized acetylcholine esterase sheet and matched matrix sheet and their preparation method and uses. The immobilized acetylcholine esterase uses qualitative filter paper as carrier and is adsorbed in filter paper. The immobilized acetylcholine esterase sheet uses crucian brain as source of acetylcholine esterase and qualitative filter paper as fixed carrier by physical adsorption method. The matched matrix sheet is prepared using the following method : mixing thymol blue alcohol solution of weight percentage 0.1-1% and 0.1-0.5mol / L -1acetylcholine according to volume ratio 1:1-1:5, titrating with 0.05-0.2mol / L-1 sodium hydroxide to make the color changing from red to blue, stop titration instantly, adding it to culture dish, filter paper is added and then matrix sheet is obtained. The inventive enzyme sheet reproduce well, has high enzyme activity. The enzyme sheet is easy to store and can be used for detecting carbamate pesticide, especially in quick agricultural pesticide residue detection.

Description

technical field [0001] The invention relates to an immobilized acetylcholinesterase tablet and a supporting matrix tablet, as well as their preparation method and application, belonging to the field of biotechnology. Background technique [0002] Among the pesticides and insecticides sold in the market, more than 50% belong to organophosphorus and carbamate insecticides, and their output accounts for more than 70%. These insecticides are popular with farmers because of their low price and the rate and degree of development of resistance are lower than pyrethroid insecticides. For the detection of pesticide quality, although there are necessary detection methods and means, most of them are limited to specific laboratories, and large-scale instruments such as gas spectrometry and liquid spectrometry are used for detection. These devices are complicated to operate, expensive, require high working conditions, and take a long time for sampling inspection. At present, the common...

AI Technical Summary

Problems solved by technology

However, this method has some shortcomings

First, the acetylcholinesterase (AchE) used is expensive; second, some pesticides do not significantly inhibit AChE due to in vitro inhibition

[0003] Compared with free enzymes, immobilized enzymes have the advantages of strong stability, good reproducibility, and reusability. Regarding making acetylcholinesterase (AchE) into enzyme tablets, and then using color reaction to carry out carbamate pesticides The research on testing is currently very active in this field, but the current research level is still limited to the production, and it cannot be used to detect pesticides