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

A technology for agricultural and sideline products and veterinary drug residues, which is applied in the preparation of test samples, measuring devices, instruments, etc., can solve the problems of narrow application range, poor specificity, and complicated operation.

Inactive Publication Date: 2006-01-04
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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034]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. Put the processed sample into a centrifuge tube, add 6 mL of ethyl acetate and shake for 10 min; centrifuge at 3000 g at room temperature for 10 min; take out 2 mL of the upper layer of ethyl acetate (equivalent to 1 g of sample) and dry it under N2 at 60 °C.

[0035] Preparation of washing liquid: Tris54g, boric acid 27.5g, dissolve with a small amount of DD-H2O, add 20ml of 0.5mol / L EDTA-Na2 (PH8.0), add DD-H2O to dilute to 1000ml. Add 200ml of saturated (NH4)2SO4 solution, 40ml of Tween-20 and 480ml of DD-H2O, and mix well.

[0036] Preparation of reagent biotinylated antibody: take 1g of biotin, suspend it in 12ml N,N dimethylformamide DMF, add 0.6...

Embodiment 2

[0043] The pretreatment of sample and the preparation of washing solution are the same as in Example 1

[0044] 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.

[0045] Reagent ...

Embodiment 3

[0049] The pretreatment of sample and the preparation of washing solution are the same as in Example 1

[0050] 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 present invention belongs to the field of toxic matter detecting technology. The detection method of quickly detecting veterinary medicinal residue in food and agriculture and sideling products includes the following steps: applying coupled antigen of several kinds of veterinary medicines as micro array on solid carrier and coating; mixing deluent diluted biotinylated antibody and measured sample, competitive immunogical reaction with the coated veterinary medicinal antigen, and washing with detergent; adding avidin marked colloidal gold for amplification and colorizing, and obtaining grey result with scanning instrument. The competitive immunogical method of the present invention has the features of high sensitivity, high specificity and simple operation.

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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IPC IPC(8): G01N33/543G01N33/531G01N1/28
Inventor 王华全林荣业
Owner YUHUAN MARINE BIOCHEM
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