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Enzyme linked immunological adsorption detection method for analyzing residuals of cyano pyrethroid pesticides

An enzyme-linked immunosorbent, pyrethroid technology, applied in the field of immunoassay, can solve the problems of difficult to achieve on-site detection, expensive instruments, cumbersome and other problems, and achieve the effects of low sample detection cost, no radioactive pollution, and less time-consuming

Inactive Publication Date: 2008-12-31
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, traditional pesticide residue analysis mainly relies on gas chromatography, liquid chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, etc. These methods require expensive instruments and require cumbersome pretreatments, making it difficult to meet the requirements of quick and easy on-site detection

Method used

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  • Enzyme linked immunological adsorption detection method for analyzing residuals of cyano pyrethroid pesticides
  • Enzyme linked immunological adsorption detection method for analyzing residuals of cyano pyrethroid pesticides
  • Enzyme linked immunological adsorption detection method for analyzing residuals of cyano pyrethroid pesticides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Operation of the detection method and calculation of results:

[0025] (1) Antigen coating

[0026] The hapten meta-phenoxybenzyl cyanohydrin succinate and ovalbumin (OVA) conjugated complex was used as the coating antigen, and the coating antigen was diluted 1:100 with 0.05M, pH 9.6 sodium carbonate buffer ~1:200, the concentration is 4~8μg / ml as the coating solution, add 100μL of the coating solution to each well of the ELISA plate, incubate overnight at 4℃, use the above sodium carbonate buffer containing 2% OVA as the blocking solution Closed

[0027] (2) Competitive reaction

[0028] The polyclonal m-phenoxybenzyl cyanohydrin succinate polyclonal antibody prepared by immunization was diluted with antibody diluent (containing 1% gelatin, containing 0.5% Tween-20, phosphate buffer solution PBST with pH=7.4) according to 1: After diluting at a ratio of 100 to 1:200, add 50μL to each well and add a series of concentrations of 0ng / mL, 0.1ng / mL, 1.0ng / mL, 10ng / mL,...

Embodiment 2

[0034] Example 2: Specificity experiment of detection method:

[0035] The cyano-containing pyrethroid was selected as the test substance, and the inhibitory concentration (IC 50 ), and then use the following formula to calculate the cross-reactivity of the antibody to these substances; the greater the cross-reaction rate, the stronger the affinity of the antibody for cyano-containing pyrethroids, and vice versa, the poorer the affinity of the antibody.

[0036] Cross reaction rate (CR%) = [IC 50 (M-phenoxybenzyl cyanohydrin succinate) / IC 50 (Containing cyano pyrethroid)]×100%.

[0037] The results of the experiment are shown in Table 1. Using indirect ELISA method, the polyclonal antibody containing m-phenoxybenzyl cyanohydrin succinate is used for the intermediate derivative m-phenoxybenzyl cyanohydrin butadiene containing cyano pyrethroid. Ester, and 8 kinds of cyano-containing pyrethroids all have cross-reactivity. However, the cross-reaction rate for non-cyano pyrethroids suc...

Embodiment 3

[0040] Example 3: Addition and recovery experiment:

[0041] (1) Addition of the sample: add the cyano pyrethroid sample to the cut cotton at 5.0ng / g, 10.0ng / g, 50.0ng / g, 100.0ng / g, and let stand at room temperature for at least 15min .

[0042] (2) Extraction and purification of samples: Take 1g of cotton samples that have been added with different concentrations of standards and place them in a 50mL centrifuge tube. Then add 30 mL of ethyl acetate saturated with acetonitrile, ultrasonically extract for 30 minutes, squeeze out the extract with a glass syringe, repeat three times, combine the extracts, blow dry with nitrogen, and add 1 mL of standard diluent as a sample for analysis. Five samples were prepared for each concentration, the content was determined, and the measured concentration was compared with the added concentration.

[0043] Calculation of recovery rate: Calculate the corresponding inhibition rate according to the OD value of the sample with different addition conc...

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Abstract

The invention discloses an enzyme-linked immunosorbent assay method for multi-residue analysis of a cyano-containing pyrethroid pesticide, belonging to the technical field of immunoassay. The enzyme-linked immunosorbent assay method uses a coupled complex of hapten m-phenoxy benzal cyanohydrin succinate ester and ovalbumin as a coating antigen, samples of the cyano-containing pyrethroid and a polyclonal antibody of the m-phenoxy benzal cyanohydrin succinate ester are added, then the coating antigen and the cyano-containing pyrethroid to be detected are competed and reacted with the antibody, an enzyme-labeled secondary antibody is added to be combined with the fixed antibody, washing liquid is used for washing, developer is added, an enzyme-labeled instrument is used for detecting OD value, the OD value is compared with a standard curve of a standard product of the cyano-containing pyrethroid, and the concentration of the cyano-containing pyrethroid of the samples to be detected is calculated. The enzyme-linked immunosorbent assay method can accurately and sensitively detect the residues of the cyano-containing pyrethroid pesticide in the samples to be detected, the pre-treatment process of the samples is simple with less time-consuming, a large number of samples can be simultaneously detected, and the detection cost of the samples is far lower than the detection method of the traditional instrument; furthermore, the method of the invention has good stability and no radioactive pollution.

Description

Technical field [0001] An enzyme-linked immunosorbent assay method for multi-residue analysis of cyano pyrethroid pesticides, specifically, it relates to an enzyme-linked immunosorbent assay suitable for multi-residue analysis of cyano pyrethroid (a-Cyano-Pyrethroid) pesticides The adsorption detection method belongs to the field of immunoassay technology. Background technique [0002] Pyrethroid is a class of biomimetic drugs developed on the basis of the research on the chemical structure of natural pyrethroids. The application of natural pyrethrum has a long history. Because of its strong knockdown power, fast insecticidal effect, broad-spectrum, easy degradation, low toxicity to higher animals and birds, safe use, and low environmental pollution, it is It is widely used for the prevention and control of pests such as tea gardens, orchards, farmland, etc. In particular, a-Cyano-Pyrethroid is more widely used. With the improvement of international standards, the internationaliz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/78
Inventor 胥传来殷祥刚郝晓蕾李雅丽袁媛
Owner JIANGNAN UNIV
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