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Test strip for detecting acetamiprid, and preparation method and application thereof

A technology of acetamiprid and test strips, which is applied in the test strips for detecting acetamiprid and its preparation and application fields, can solve the problems of complex and expensive instruments and equipment, cumbersome and time-consuming processing, and long detection time, and achieve easy promotion and use , short detection time, simple storage effect

Active Publication Date: 2018-12-21
ZHENGZHOU TOBACCO RES INST OF CNTC +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the instrumental analysis method can achieve accurate qualitative and quantitative analysis, it requires complex and expensive instruments and professional operators. In addition, the sample pretreatment is cumbersome and time-consuming, the detection time is long, and the detection cost is high, which has limitations for the development of on-site detection work.

Method used

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  • Test strip for detecting acetamiprid, and preparation method and application thereof
  • Test strip for detecting acetamiprid, and preparation method and application thereof
  • Test strip for detecting acetamiprid, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation of test strips for detecting acetamiprid

[0044] The preparation method of this test strip mainly comprises the following steps:

[0045] 1) Prepare a conjugate release pad sprayed with acetamiprid monoclonal antibody-colloidal gold marker;

[0046] 2) Prepare a reaction membrane with a detection line coated with acetamiprid hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;

[0047] 3) Assemble the conjugate release pad prepared in 1) and 2), the reaction membrane, the sample absorption pad, the water absorption pad and the PVC bottom plate into a test strip.

[0048] The following step-by-step detailed description:

[0049] 1. Synthesis and identification of acetamiprid hapten

[0050] Take 0.50 g of N-desmethyl acetamiprid, add 50 mL of methanol to dissolve, add 0.30 g of aminobutyric acid, stir, add 0.37 mL of 37% formaldehyde solution, stir, mix well, react at 80°C for 4 h; stop the react...

Embodiment 2

[0092] The detection of acetamiprid in the sample of embodiment 2

[0093] 1. Sample pretreatment

[0094] Weigh 1.0g±0.05g of crushed tobacco leaf sample into a polystyrene centrifuge tube; add 5 mL of 50% methanol aqueous solution, and use a homogenizer to fully break it up to obtain a sample liquid; pipette 100 μL of sample liquid and 900 μL of The PBS buffer was mixed and tested.

[0095] 2. Test with test strips

[0096] Use a pipette to take 70 μL of the sample solution to be tested and drop it vertically into the sample hole, start timing when the liquid flows, react for 10 minutes, and judge the result.

[0097] 3. Analyze test results

[0098] Negative (-): The color of the T line is darker than that of the C line or consistent with the color of the C line, indicating that the concentration of acetamiprid in the sample is lower than the detection limit, such as figure 2 a, 2b.

[0099] Positive (+): The color of the T line is lighter than that of the C line or th...

Embodiment 3

[0101] Embodiment 3 sample detection example

[0102] 1. Detection limit test

[0103] Take blank tobacco leaf samples, add acetamiprid to the final concentrations of 1.5, 3, and 6 mg / kg, respectively, take test strips for detection, and repeat the measurement three times for each sample.

[0104] When testing tobacco leaf samples with test strips, when the concentration of acetamiprid is 1.5 mg / kg, the test strip shows that the T line color is darker than the C line color or is consistent with the C line color, which is negative; When the concentration of acetamiprid added is 3 or 6 mg / kg, the test strip shows that the color of the T line is lighter than that of the C line or the T line is not colored, which is positive, indicating that the test strip has no effect on tobacco leaves. The detection limit of acetamiprid was 3 mg / kg.

[0105] 2. False positive rate and false negative rate test

[0106] Take 20 positive tobacco leaf samples known to have acetamiprid content gr...

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Abstract

The invention provides a test strip for detecting acetamiprid, and a preparation method and application thereof and further provides a method using the test strip to detect the acetamiprid. The test strip comprises a sample absorbing cushion, a conjugate releasing cushion, a reacting film, a water absorbing cushion and a base plate. The reacting film is provided with detection lines coated with acetamipridhatpin-carrier protein couplers and quality control lines coated with anti-goat and anti-mouse anti-bodies, and acetamipridmonoclonal antibody-colloid gold markers are sprayed on the conjugate releasing cushion. Acetamiprid monoclonal antibodies are prepared with the acetamiprid hatpin-carrier protein couplers as immunogen, the acetamiprid hatpin-carrier protein couplers are prepared through the method that acetamiprid hatpin and acarrier protein are coupled, and the acetamiprid hatpin is obtained through the reaction of N-nor-acetamiprid and aminobutyric acid. The test strip and thedetection method have the advantages that operation is easy, the sensitivity is high, the detecting speed is high, the cost is low, limitation from detecting equipment is avoided, and can conduct rapid detection and on-site monitoring on the acetamiprid in great batches of samples.

Description

technical field [0001] The invention relates to a test strip for detecting acetamiprid and its preparation method and application, in particular to a colloidal gold test strip for detecting acetamiprid, which is especially suitable for rapid detection of acetamiprid in tobacco. Background technique [0002] Acetamiprid belongs to neonicotinoid insecticides, which are an important class of insecticides after organophosphorus, carbamate and pyrethroid insecticides. By selectively controlling the nicotinic acetylcholinesterase receptors (nAChRs) in the central nervous system of insects, the normal conduction of the central nervous system of insects is blocked, resulting in general paralysis and death of the pests. Acetamiprid is a representative agent of neonicotinoid insecticides. It is widely used in agricultural production to prevent and control various pests in seeds, leaves and soil. The effect of pest control is ideal, and it is mainly used for the control of tobacco aph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/58G01N33/577G01N33/531
CPCG01N33/531G01N33/558G01N33/577G01N33/58
Inventor 范子彦陈黎万宇平崔海峰刘惠民唐纲岭潘立宁颜权平冯才伟
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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