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Enzyme biosensor for pesticide residue detection and preparation method and application of enzyme biosensor

A biosensor and product technology, applied in the field of biosensing, can solve the problems of difficult real-time and rapid on-site detection, expensive equipment, time-consuming and other problems, and achieve the effects of fast and accurate identification and measurement, low price and good conductivity

Inactive Publication Date: 2019-04-12
武汉中科志康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection results of large-scale instruments are accurate, reliable, and highly sensitive, but there are disadvantages of expensive equipment, long time-consuming, high cost, and the need for professional and technical personnel, making it difficult to perform rapid on-site detection in real time

Method used

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  • Enzyme biosensor for pesticide residue detection and preparation method and application of enzyme biosensor
  • Enzyme biosensor for pesticide residue detection and preparation method and application of enzyme biosensor
  • Enzyme biosensor for pesticide residue detection and preparation method and application of enzyme biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A nano-gold tungsten disulfide graphene composite material AuNPS / WS 2 -GR, its preparation method is as follows:

[0032] 1) Add 0.05g graphene oxide powder GO into 40mL methanol, ultrasonically disperse for 1 hour to obtain a uniformly dispersed graphene oxide methanol solution, then slowly add 0.5g sodium tungstate powder and 0.5g thiourea to the In the methanol solution of graphene oxide, sonicate for 30 minutes to obtain a mixed solution. Subsequently, the mixed solution was transferred to a 100 mL autoclave, and reacted at 180° C. for 64 hours. After the reaction, the reactor was naturally cooled, and the resulting solution was filtered to obtain a black product. The product was washed 3 times with twice distilled water, and the product was dried at 60°C under nitrogen protection to obtain WS 2 Modified graphene WS 2 -GR, spare.

[0033] 2) 20mL 0.2%HAuCl 4 solution and 1g cetylpyridinium bromide were sequentially added to a 50mL centrifuge tube, ultrasonical...

Embodiment 2

[0036] An Enzyme Biosensor Plant-E / AuNPS / WS for Detecting Pesticide Residues 2 -GR / SPE, its preparation method is as follows:

[0037] 1) 1mg of the prepared graphene composite material AuNPS / WS 2 -GR dispersed in 1 mL of secondary water to obtain AuNPS / WS 2 -GR aqueous solution, use a pipette gun to pipette 5 μL of the dispersion solution and drop it on the working electrode of the screen-printed electrode, dry it at room temperature, and use 0.01mol•L -1 Washing with PBS buffer (pH=7.4) to obtain graphene composite modified electrode AuNPS / WS 2 -GR / SPE.

[0038] 2) Prepare plant esterase Plant-E to a concentration of 100U•mL -1 The aqueous solution, the aqueous solution and the chitosan solution with a mass concentration of 0.3% were mixed in equal volumes to form a mixed solution, and 5 μL of the mixed solution was drip-coated on the AuNPS / WS prepared in step 1). 2 - On the working electrode of GR / SPE, dry at room temperature, use 0.01mol•L -1 PBS buffer (pH=7.4) wash...

Embodiment 3

[0041] Application of an enzyme biosensor for detecting pesticide residues in the detection of pesticide residues. The detection object is organophosphorus pesticides (parathion or malathion). The constructed plant esterase electrochemical biosensor is used for detection. The detection principle: The substrate 1-naphthyl acetate is hydrolyzed by plant esterase to produce acetic acid and naphthol (electroactive substances), thereby generating current signals. The inhibition of plant esterase activity by organophosphorus pesticides reduces the electroactive products, so the current signal decreases. Depending on the increase in the content of organophosphorus pesticides, the decrease in the current signal is also different (the detection principle is as follows: figure 2 ). The method is as follows:

[0042] 1) Detection of blank samples: the prepared Plant-E / AuNPS / WS 2 - The GR / SPE enzyme biosensor was immersed in a phosphate buffer solution without organophosphorus pesticid...

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Abstract

The invention relates to the technical field of biosensing and the field of pesticide residue detection, and discloses an enzyme biosensor for pesticide residue detection and a preparation method of the enzyme biosensor and application of the enzyme biosensor in rapid detection of organophosphorus pesticide residues. An enzyme inhibition principle is adopted, the surface of a ready-to-use type screen-printed electrode is successively modified with a graphene composite material and plant esterase to construct the enzyme biosensor. The differential-pulse voltammetry is adopted to construct a standard curve to achieve simple, rapid and highly-sensitive detection of organophosphorus pesticide residues. The adopted ready-to-use type screen-printed electrode is free of manual grinding and is simple to operate, and detection repeatability is improved. The graphene composite material with a large specific surface area and good conductivity is used for modifying the electrode, and therefore thefixed amount of the enzyme is increased, electron transfer in the working process of the sensor is facilitated, and limit of detection and sensitivity are obviously improved. The adopted plant esterase is wide in source and low in cost, and therefore the detection cost is lowered.

Description

technical field [0001] The invention belongs to the technical field of biosensing and the field of pesticide residue detection, and specifically relates to a synthesis method of a graphene composite material, and an enzyme biological agent for detecting pesticide residues constructed from the graphene composite material, plant esterase and screen printing electrodes. Fabrication methods and applications of sensors. Background technique [0002] Pesticides are chemical agents used to prevent and control plant diseases and insect pests and regulate plant growth. Their use has become the guarantee for high-quality and high-yield crops in most agricultural areas. However, the large-scale application of pesticides will also bring huge negative impacts on the ecological environment and human survival. A series of serious ecological, environmental, and food safety problems caused by undegraded pesticide residues in the soil, air, and water environments have become global concerns....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3272
Inventor 高金娥
Owner 武汉中科志康生物科技有限公司
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