Detection kit for detecting sulfadimidine in food

A technology of sulfamethazine and a detection kit, which is applied in the field of detection kits for sulfamethazine in food, can solve the problems of easy inactivation of marker enzymes, complex environmental interference factors, and qualitative detection only. Achieve high sensitivity and specificity

Inactive Publication Date: 2017-05-31
BEOSON JIANGSU FOOD SAFETY TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA method undergoes multiple elution processes, the marker enzyme is easily inactivated, the substrate is easily decomposed when exposed to light, and the environmental interference factors are complicated.
Colloidal gold chromatography can meet the requirements of on-site rapidity, but only qualitative detection can not obtain quantitative results

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: Preparation of specific components of the kit

[0027] 1. Synthesis of sulfamethazine hapten

[0028] A mixture of 0.58 g sulfamethazine and 0.082 g ethylenediamine in 80 ml methanol was reacted at room temperature for 4 to 5 hours, and the solvent was evaporated to obtain the sulfamethazine hapten quantitatively.

[0029] 2. Preparation of enzyme-labeled antigen

[0030] Take 10~15mg sulfamethazine hapten, dissolve it in 1~1.5ml N,N-dimethylformamide (DMF); take 30~35mg dichloroethane (EDC) and N-hydroxysuccinyl Amine (NHS) was fully dissolved in 0.2-0.3ml of water, then added to the hapten solution, and stirred at room temperature for 20 hours to obtain reaction solution A; weigh 35-45 mg of horseradish peroxidase (HRP) to make it Fully dissolve in pH 7.2, 4ml phosphate buffer solution, slowly add reaction solution A dropwise into HRP solution, and stir at room temperature for 20h; dialyze with 0.015mol / L phosphate buffer solution at 4°C For 3 days, t...

Embodiment 2

[0043] Embodiment 2: the formation of kit

[0044] Set up the detection kit of sulfamethazine in food so that it contains the following components:

[0045] Horseradish peroxidase-labeled sulfamethazine hapten marker

[0046] Enzyme-labeled Antigen Diluent

[0047] Conjugates of sulfamethazine monoclonal antibody and magnetic beads

[0048] Magnetic Label Antibody Diluent

[0049] Sulfamethazine series standard solutions, concentrations are: 0μg / L, 0.025μg / L, 0.05μg / L, 0.1μg / L, 0.2μg / L, 0.4μg / L, 0.8μg / L, 1.6μg / L, the standard diluent is phosphate buffer containing 0.035wt.% Tween-20, 0.042mol / L pH value 7.4.

[0050] Concentrated complex solution is 20 times concentrated complex solution, specifically containing 45-60g of NaH per liter 2 PO 4 2H 2 O, 200~220g Na 2 HPO 4 12H 2 Aqueous solution of O.

[0051] The concentrated washing solution is a 20-fold concentrated washing solution, specifically a phosphate buffer solution with a pH value of 7.2-7.4 containing 0.3...

Embodiment 3

[0052] Embodiment 3: the detection of residual amount of sulfamethazine in the sample

[0053] 1. Sample pretreatment method

[0054] (1) milk

[0055] Take 25 μL of fresh milk sample and add 950 μL of sample diluent (dilute the concentrated complex solution to 10 times the volume with deionized water), vortex and mix well, and take the solution for sample analysis.

[0056] (2) milk powder

[0057] Weigh 0.5g±0.05g milk powder sample, add 5ml sample diluent, vortex mix, take 200μL from it and add to 600μL sample diluent, vortex mix, take the solution for sample analysis.

[0058] 2. Detection with kit and analysis of results

[0059] Dilute the enzyme-labeled antigen and the enzyme-labeled antigen diluent at a volume ratio of 1:20 to obtain the enzyme-labeled antigen working solution; dilute the magnetic-labeled antibody and the magnetic-labeled antibody diluent at a volume ratio of 1:20 to obtain the magnetic-labeled antibody Antibody working solution: Take 30-35 μL of e...

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Abstract

The invention discloses a detection kit for detecting sulfadimidine in food. The detection kit is prepared from enzyme-labeled antigen, enzyme-labeled antigen diluent, a magnetic-labeled antibody, magnetic-labeled antibody diluent, a sulfadimidine series standard substance solution, chemiluminescent substrate A liquid, chemiluminescent substrate B liquid, a concentrated complex solution and concentrated washing liquid. The detection kit provided by the invention has higher sensitivity and specificity; the detection sensitivity to the sulfadimidine can reach 0.025mu g/L.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a detection kit for sulfamethazine in food. Background technique [0002] Sulfamethazine (SMZ) is a broad-spectrum antibacterial agent that has inhibitory effects on most Gram-positive and Gram-negative bacteria. It is mainly used clinically to treat various hemolytic streptococci and meningitis Bacterial diseases such as respiratory tract infection and intestinal infection caused by coccus, pneumococcus and other infections are widely used in animal husbandry, and the residue phenomenon in animal food is relatively serious. Because sulfamethazine has a long time of action and metabolism in the body, sulfamethazine ingested through any route may accumulate in the human body. When it exceeds a certain concentration, it will cause damage to the human body. Long-term or excessive use It can cause damage to the urinary system, hematopoietic system, nervous system, etc., and e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543G01N33/577
CPCG01N33/535G01N33/54326G01N33/577G01N33/9446G01N2430/00
Inventor 周朱晨张根义胡彬张进吴念绮周合
Owner BEOSON JIANGSU FOOD SAFETY TECH CO LTD
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