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Method and device for detecting pesticide

A technology of pesticides and magnetic nanoparticles, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems affecting measurement

Inactive Publication Date: 2012-07-18
SONY CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the inhibited acetylcholinesterase can be partially revived by oxime (such as prolidoxime iodide) to retain the activity of acetylcholinesterase, the fixed layer of acetylcholinesterase on the electrode still needs to be replaced frequently to ensure the measurement Reliability, while the replacement of the fixed layer requires careful operation, otherwise the acetylcholinesterase layer will lose contact with the electrode, thereby affecting the measurement

Method used

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  • Method and device for detecting pesticide
  • Method and device for detecting pesticide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0193] Embodiment 1: Preparation of magnetic nanoparticles (MNPs)

[0194] This example provides an embodiment for the preparation of magnetic nanoparticles, i.e., the synthesis of magnetic nanoparticles via the co-precipitation route, as described in the following method:

[0195] 1) 0.98g (4mM) FeCl 3 ·6H 2 O, 0.556g (2mM) FeSO 4 ·7H 2 O and 1.152g (6mM) citric acid were dissolved in 75ml deionized (DI) water and loaded into a 250ml three-neck flask.

[0196] 2) The above solution was heated in water baths at different temperatures (0, 20, 40, 60, 80 and 100° C.), degassed in vacuum for 0.5 h, and then input argon (Ar).

[0197] 3) Dissolve 4 g (0.1 M) NaOH in 25 ml deionized water and apply ultrasound to remove soluble O 2 .

[0198] 4) Keep the NaOH solution in a water bath for 0.5h, and then quickly inject it into a three-necked bottle (containing Fe salt).

[0199]5) The reaction was held for 1 h, then naturally heated / cooled to RT.

[0200] 6) Collect the obtain...

Embodiment 2

[0202] Example 2: Immobilization of acetylcholinesterase on magnetic nanoparticles

[0203] This example provides an embodiment of the immobilization of acetylcholinesterase to magnetic nanoparticles.

[0204] Add 2ml of the magnetic nanoparticle solution (1.93g / L) in step 6) in Example 1 to a 5ml centrifuge tube, and obtain magnetic nanoparticle powder by centrifuging at 14000RPM for 30min. The obtained magnetic nanoparticle powder was washed with DI water and re-centrifuged several times. A 1 ml solution of acetylcholinesterase solution (1 U / ml) containing 1 mg / ml BSA was added to the tube and stored at 4°C for 20 h. Magnetic nanoparticles immobilized with acetylcholinesterase were obtained by magnetic separation, and washed with 4 ml of PBS one time to remove free acetylcholinesterase adsorbed on the surface of the magnetic nanoparticles. Magnetic nanoparticles immobilized with acetylcholinesterase were redispersed in 2 ml PBS and stored at 4 °C before use. The amount of...

Embodiment 3

[0205] Embodiment 3: The ratio of the cholinesterase added and magnetic nanoparticle is to the influence of enzyme immobilization amount

[0206] In this example, the effect of the ratio of cholinesterase to magnetic nanoparticles on the amount of enzyme immobilization during the immobilization of cholinesterase was studied.

[0207] Figure 13 In a certain amount of magnetic nanoparticle solution, the relationship between the amount of acetylcholinesterase added during immobilization and the actual immobilized amount is shown (n=3). Such as Figure 13 As shown, the range of enzyme immobilization that can be achieved is 0-330U / g particles. When the amount of enzyme added is 0.2ml (1U / mL), the amount of enzyme fixed on the magnetic nanoparticles after fixation is only 20.1U / g, and the immobilization efficiency is 35.8% (fixed amount / actual amount added); increase, the immobilization amount and immobilization efficiency all increase rapidly, when the amount of enzyme added is...

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Abstract

The invention relates to a method and a device for detecting pesticide by using cholinesterase. More specifically, the invention relates to a novel pesticide detection method based on acetylcholinesterase-fixed magnetic nanoparticles, wherein the magnetic nanoparticles serve as carriers to collect the acetylcholinesterase in solution. The invention also relates to a portable pen-shaped detector adopting the method for detecting pesticide.

Description

technical field [0001] The present invention relates to methods and devices for detecting pesticides using cholinesterase. Specifically, the present invention relates to a novel pesticide detection method based on magnetic nanoparticles immobilized with acetylcholinesterase (AChE), wherein the magnetic nanoparticles serve as carriers to collect cholinesterase in solution. The present invention also relates to a portable detector for pesticide detection using the above method. Background technique [0002] According to statistics, the world consumes about 2.5 million tons of pesticides every year, most of which are applied to crops. However, more than 95% of pesticides end up in water, soil and air under the exposure of rain and sunlight, causing serious environmental pollution. On the other hand, pesticide residues on crops can also cause serious food poisoning incidents. The World Health Organization (WHO) estimates that 1 million cases of pesticide poisoning occur globa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/46
Inventor 丁宁梶浦尚志李勇明
Owner SONY CORP
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