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Detection reagent kit for pesticide residue, its preparing method and its use

A technology for detecting kits and pesticide residues, applied in biochemical equipment and methods, testing food, material inspection products, etc.

Inactive Publication Date: 2004-07-14
高希武
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, the key problem of acetylcholinesterase, which is currently used in the pesticide residue rapid detection kit, is how to improve the sensitivity of the enzyme and prolong the normal temperature storage time of the enzyme.

Method used

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  • Detection reagent kit for pesticide residue, its preparing method and its use
  • Detection reagent kit for pesticide residue, its preparing method and its use
  • Detection reagent kit for pesticide residue, its preparing method and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The head of the sensitive housefly was frozen in liquid nitrogen and separated, and placed in a homogenizer at a ratio of 1:5 between the head of the housefly and the homogenate (pH 7.5, 0.1 mol / L). After the homogenate was subjected to differential centrifugation at 1000 g, 10000 g and 100000 g, the supernatant was precipitated with ammonium sulfate (50-60% saturation), and then desalted with a G25 column. After desalination, freeze-dry to make dry powder. Dissolve the dry powder in the buffer (pH7.5, 0.1mol / L) (w / v: 1:1) and pass it through the G200 chromatography column and the affinity chromatography column, collect the active eluate and freeze-dry to obtain the dry powder, both It is acetylcholinesterase enzyme powder. Mix the enzyme powder and the protective agent (sucrose: cyclodextrin: mannitol = 1:4:5) at a ratio of 1:100 (w / w), and after immobilization, it is the enzyme source (A) of the detection kit; as above Methods Substrate and chromogenic reagent were ...

Embodiment 2

[0032] The head of the sensitive housefly was frozen in liquid nitrogen and separated, and placed in a homogenizer at a ratio of 1:5 between the head of the housefly and the homogenate (pH 7.5, 0.1 mol / L). After the homogenate was subjected to differential centrifugation at 1,000 g, 10,000 g, and 100,000 g, the supernatant was removed and ammonium sulfate was used for precipitation (50-60 saturation), and G25 column was used for desalting. After desalination, freeze-dry to make dry powder. Dissolve the dry powder in distilled water (w / v: 1:1), centrifuge through the sucrose density gradient, collect the components of the 10%-20% sucrose gradient section, pass through the G200 chromatography column, collect the active eluent and freeze-dry it to obtain Dry powder, both acetylcholinesterase enzyme powder. Mix the enzyme powder and protective agent (starch: cyclodextrin: mannitol = 1:5:5) at 1:150 (w / w), and after immobilization, it is the enzyme source (A) of the detection kit;...

Embodiment 3

[0048] The kit prepared according to Example 1 was stored at 20-30° C. in the dark for three weeks, and the enzyme activity at each time point was measured by conventional methods. The unit of enzyme activity value is ΔOD412 value / 3 minutes.

[0049] time (days)

[0050] As can be seen from the above Table 3, the acetylcholinesterase in the kit of the present invention can be stored at room temperature for three weeks without affecting its enzyme activity. Therefore, the detection of pesticide residues can be carried out at room temperature.

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Abstract

The reagent kit for detecting pesticide residue includes acetylcholinesterase, protecting agent, primer and developing agent. The single or purified monopolymer acetylcholinesterase highly sensitive to pesticide is obtained from sensitive housefly line head. The reagent kit may be maintained at normal temperature and has high sensitivity and high measurement accuracy in detecting pesticide residue in vegetable and fruits.

Description

【Technical field】 [0001] The invention relates to a pesticide residue detection kit, which comprises acetylcholinesterase, a protective agent, a substrate and / and a chromogenic agent, wherein the acetylcholinesterase is obtained from the head of the housefly strain in a single molecular type or purified monomeric body acetylcholinesterase. 【technical background】 [0002] At present, pesticides are widely used in the process of planting food, vegetables, and fruits. Agricultural products inevitably contain trace amounts of pesticides, and it is self-evident that pesticides are harmful to people's health. With the improvement of living standards, people pay more and more attention to food safety. Therefore, the problem of pesticide residues on agricultural products is increasingly concerned by people. [0003] It is impossible to perform instrumental analysis on every vegetable and fruit sample. Therefore, there is an urgent need for a method for rapid primary screening of a ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44G01N21/25G01N33/02
Inventor 高希武
Owner 高希武
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