Reagent strip for rapidly detecting colloidal gold for 2,4-D residual

A technology of colloidal gold test paper and test strip, which is applied in the field of immunoassay, can solve the problems of complex operation, low sensitivity, and unstable test results, and achieve the effects of less time-consuming, simple pre-treatment process, and low cost of sample testing

Inactive Publication Date: 2009-06-10
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the shortcomings of the current instrumental analysis methods for pesticide residues such as high cost, complicated operation, poor specificity, low sensitivity and unstable detection results, the present invention provides a high-specificity, high-sensitivity, high-accuracy, high-precision, high-operation The method is simple and can be used for rapid detection of 2,4-D residual colloidal gold test paper in large quantities of samples

Method used

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  • Reagent strip for rapidly detecting colloidal gold for 2,4-D residual
  • Reagent strip for rapidly detecting colloidal gold for 2,4-D residual
  • Reagent strip for rapidly detecting colloidal gold for 2,4-D residual

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 2,4-D is coupled with carrier protein:

[0018] (1) Coupling of 2,4-D and bovine serum albumin (BSA) by active ester method

[0019] Weigh 250mg 2,4-D (about 270umol) and dissolve it in 10mL anhydrous dimethylamide (DMF) (A solution), weigh 500mg BSA and dissolve it in 10mL 0.01mol / L pH7.4 PBS (B solution), Dissolve 250mg of EDC in an appropriate amount of PBS (solution C), drop liquid A into liquid C and react for 1 hour; then drop the reaction liquid into liquid B drop by drop, and stir and react at room temperature for 12 hours, then dialyze and centrifuge the reaction liquid, and The supernatant was freeze-dried and stored at 4°C.

[0020] (2) Mixed acid anhydride method coupling 2,4-D and ovalbumin (OVA)

[0021] Accurately weigh 250 mg of 2,4-D, add 10 mL of DMF to dissolve, then add 50.0 μL of tri-n-butylamine and 30 μL of isobutyl chlorobutyrate to this solution in sequence, and magnetically stir overnight at room temperature; Add dropwise to 20mL 0...

Embodiment 2

[0022] Example 2 Preparation of 2,4-D polyclonal antibody:

[0023] The conjugate of 2,4-D and BSA was used as the immunogen, and 2 mg of the immunogen was dissolved in 1 mL of sterile saline, mixed with 1 mL of complete Freund's adjuvant, fully emulsified, and injected subcutaneously on the neck and back of New Zealand white rabbits (6 After that, boost the immunization once every two weeks, and use incomplete Freund's adjuvant to mix with the immunogen. The immunization site is the same. From the third immunization, blood is collected from the rabbit ear vein one week after each immunization to detect the serum titer . A total of 5 times of immunization, the whole blood was collected from the rabbit carotid artery one week after the last immunization, the rabbit antiserum was crudely extracted by octanoic acid-saturated ammonium sulfate method, and then further purified by DE-52 anion exchange chromatography to obtain relatively pure 2 , 4-D polyclonal antibody.

Embodiment 3

[0024] Example 3 Preparation of Colloidal Gold

[0025] Preparation of colloidal gold by improved method of sodium citrate

[0026] (1) Prepare 100mL of 0.01% chloroauric acid with newly prepared deionized water, place it in a conical flask, and heat it in an electric furnace until it boils.

[0027] (2) Accurately absorb a certain amount of 1.0% trisodium citrate, quickly add it to the Erlenmeyer flask, shake it evenly, immediately put it into the electric furnace and continue boiling until the chloroauric acid turns red, then take it out. After cooling to room temperature, restore to the original volume with deionized water, and store in a sterile airtight seal. Such as figure 2 The maximum absorption peak wavelength of the 400-700nm ultraviolet scanning shown is 525nm, and there is a smaller absorption peak at 237nm, and the double peak is the characteristic of colloidal gold. As shown in Figure 2 and Figure 3, the size of the colloidal gold particles observed under the...

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Abstract

The invention relates to a fast detection colloidal gold test strip used for 2, 4-D residual, belonging to the technical field of immunity analysis. The invention overcomes the defects of complex operation, high detection cost, slow speed and secondary pollution and the like in the 2,4-D physical and chemical analysis, and fast and accurately detects 2, 4-D residual in garden spgarden stuff, cereal crops and water samples. During the detection process, 2,4-D in the sample is reacted with specificity of gold marking 2,4-D antibody (quantification), 2, 4-D content in the sample is determined (coloration is feminine, namely not exceeding controlling quantity; otherwise exceeding the controlling quantity) through specificity fixation reaction of peridium 2, 4-D envelope antigen (detection line, T line) on the test strip cellulose acetate; pre-custodite goat anti-rabbit antibody on the test strip cellulose acetate is used as a quality control line (C line, coloration is effective, otherwise ineffective). Shelf life of the test strip is at least six months.

Description

1. Technical field [0001] The invention relates to a colloidal gold test strip for detecting 2,4-D residues, which is mainly suitable for the rapid determination of 2,4-D residues in samples such as fruits and vegetables, water and grains. The invention belongs to the technical field of immune analysis. 2. Background technology [0002] 2,4-dichlorophenoxyacetic acid (2,4-dichlorophenoxyacetic acid, 2,4-D) is a phenoxycarboxylic acid herbicide and plant growth regulator, widely used in wheat, barley, grain, lawn There are about 1,500 kinds of pesticides that contain 2,4-D active ingredients. The acute toxicity of 2,4-D is mainly neurotoxicity; 2,4-D can also stimulate the gastrointestinal tract and cause nausea, vomiting and diarrhea; skin contact with 2,4-D may cause rash or dermatitis. It is also reported that 2,4-D may have potential "triple-induced" effects. Due to its extensive use in agricultural production, it has caused pollution of fruits and vegetables, grain, g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/52
Inventor 曹远银于基成王菡
Owner SHENYANG AGRI UNIV
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