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Detecting method for quick detecting veterinary medicinal residue in food and agriculture and sideline products

A technology for residues of agricultural and sideline products and veterinary drugs, which is applied in the detection of veterinary drug residues in food and in the field of detection of agricultural and sideline products. short time effect

Inactive Publication Date: 2009-07-22
YUHUAN MARINE BIOCHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the shortcomings of poor specificity, low sensitivity, narrow application range and complicated operation in the detection method used in the prior art, and provides a method for detecting veterinary drug residues with simple sample pretreatment, simple detection, rapid, sensitive and accurate detection

Method used

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  • Detecting method for quick detecting veterinary medicinal residue in food and agriculture and sideline products
  • Detecting method for quick detecting veterinary medicinal residue in food and agriculture and sideline products
  • Detecting method for quick detecting veterinary medicinal residue in food and agriculture and sideline products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Pretreatment of honey samples: Take 1g of honey, put it into a centrifuge tube, add 2mL of distilled water to dissolve; add 2mL of ethyl acetate to shake for 10min; centrifuge at 3000g at room temperature for 10min; In a test tube, blow dry with N2 at 60°C; dissolve the residue with 0.5mL buffer 1; take 50uL for analysis.

[0044] Shrimp sample pretreatment: shell the shrimp sample, homogenize the sample with a homogenizer; take 3g of the homogenized sample, put it into a centrifuge tube, add 6mL of ethyl acetate to shake for 10min; centrifuge at 3000g at room temperature for 10min; take out 2mL of the upper layer of acetic acid Ethyl ester (equivalent to 1 g sample) was dried under N2 at 60°C.

[0045] Preparation of washing solution: 54g Tris, 27.5g boric acid, dissolve with a small amount of DD-H2O, add 20ml 0.5mol / L EDTA-Na2 (PH 8.0), add DD-H2O to make the volume to 1000ml. Add 200ml of saturated (NH4)2SO4 solution, 40ml of Tween-20 and 480ml of DD-H2O, and mix we...

Embodiment 2

[0055] The pretreatment of sample and the preparation of washing solution are the same as in Example 1

[0056] Preparation of reagent biotinylated antibody: Take 1g of biotin, suspend it in 12ml N,N dimethylformamide DMF, add 0.6g N-hydroxysuccinimide HOSU and 0.8g dicyclohexylcarbodiimide DCC was placed in an airtight container and magnetically stirred overnight at room temperature. The filtrate was spun to dryness. Add 10ml of diethyl ether for washing, and then add 200ml of isopropanol for crystallization to obtain white powdery activated biotin crystals. Use DMF to make BNHS into a 1mg / mL solution; dilute the antibody to be labeled with 0.10mol / L sodium bicarbonate solution with a pH value of 9.0 to 2mg / ml, and after dialysis, use 1mg / ml BNHS: antibody to be labeled = 1 : 10 and mix with occasional shaking for 3 hours. Add 0.05mol / L, pH7.4 PBS solution into the dialysis bag, dialyze overnight at 4°C, and dilute to the working concentration when used.

[0057] Reagent ...

Embodiment 3

[0061] The pretreatment of sample and the preparation of washing solution are the same as in Example 1

[0062] Preparation of reagent biotinylated antibody: take 1g of biotin, suspend it in 12ml N,N dimethylformamide DMF, add 0.6g N-hydroxysuccinimide HOSU and 0.8g dicyclohexylcarbodiimide DCC , placed in an airtight container, and magnetically stirred overnight at room temperature. The filtrate was spun to dryness. Add 10ml of diethyl ether for washing, and then add 200ml of isopropanol for crystallization to obtain white powdery activated biotin crystals. Use DMF to make BNHS into a 1.2 mg / mL solution; dilute the antibody to be labeled with 0.12 mol / L sodium bicarbonate solution with a pH value of 9.2 to 32 mg / ml, and after dialysis, use 1 mg / ml BNHS: Antibody to be labeled = Mix at a ratio of 1:15 with occasional shaking for 5 hours. Add 0.1mol / L, pH7.6 PBS solution into the dialysis bag, dialyze overnight at 4°C, and dilute to the working concentration when used.

[0...

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Abstract

The invention provides a method for detecting veterinary drug residues in agricultural by-products and food, and belongs to the technical field of toxic substance detection. The detection method for rapid detection of veterinary drug residues in food and agricultural by-products comprises the following steps: spotting multiple veterinary drug-coupled antigens on a solid-phase carrier according to a microarray and coating it; wherein the solid-phase substrate is immobilized with veterinary drug antigens ; Mix the biotinylated antibody diluted with the diluent and the sample to be tested, and perform a competitive immunoreaction with a variety of veterinary drug-coated antigens immobilized on the solid phase carrier, and wash with the washing solution; add avidin-labeled colloidal gold for amplification and stained, grayscale results obtained by scanner. Compared with the detection of the prior art, the invention adopts the immunocompetition method to detect, and mainly has the characteristics of high sensitivity, strong specificity and easy operation. The immunocompetitive method requires fewer reagents, simple sample addition procedure, and short reaction time; at the same time, measurement and data processing are easy to automate.

Description

technical field [0001] The invention belongs to the technical field of toxic substance detection, and in particular relates to a detection method for detecting veterinary drug residues in agricultural by-products and food. Background technique [0002] my country is a big country in the production and consumption of agricultural products, and the detection of veterinary drug residues in agricultural products is directly related to the production, export and food consumption safety of agricultural products in my country. In particular, the export of agricultural products in my country has continuously encountered foreign technical barriers to trade due to veterinary drug residues in recent years. Only Zhejiang Province’s agricultural product exports have been banned for hundreds of millions of dollars a year. In recent years, food safety issues have become one of the key issues of national concern. Food pollution incidents and export obstructions caused by toxic residues exce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531G01N1/28
Inventor 王华全林荣业
Owner YUHUAN MARINE BIOCHEM
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