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Colloidal gold kit for detecting beta-stimulant drug and preparation method thereof

A stimulant and colloidal gold technology, which is applied in the colloidal gold kit for detecting β-stimulant drugs and its preparation field, can solve the problem of low sensitivity of capillary electrophoresis, low stability of enzyme-linked immunosorbent method, and chromatographic operation process. cumbersome and other problems, to achieve the effect of improved sensitivity, low cost, and fast chromatography

Active Publication Date: 2018-04-20
杭州南开日新生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection methods of β-agonist drugs in raw milk and fresh milk mainly include ELISA, capillary electrophoresis, chromatography, etc. These detection methods have various defects, such as the low stability of ELISA , because enzymes are easily affected by the environment, the time is long, special instruments are required, and the cost is high; capillary electrophoresis has low sensitivity and long analysis time; High; therefore, in practice, it is necessary to establish a detection method with high sensitivity, simple operation, low cost, various types of detection drugs, and suitable for large-scale promotion

Method used

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  • Colloidal gold kit for detecting beta-stimulant drug and preparation method thereof
  • Colloidal gold kit for detecting beta-stimulant drug and preparation method thereof
  • Colloidal gold kit for detecting beta-stimulant drug and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 A colloidal gold kit for detecting β-agonist drugs

[0033] 1) Preparation of colloidal gold solution: take 1% chloroauric acid solution and dilute it 100 times with double distilled water, heat, stir and boil, add 1% trisodium citrate, continue heating and stirring until the color of the solution turns red, stop heating, and cool to room temperature to obtain a colloidal gold solution.

[0034] 2) Preparation of colloidal gold markers: Add β-agonist polyclonal antibody and ractopamine monoclonal antibody to the colloidal gold solution while stirring, then add 5% bovine serum albumin, centrifuge for 15min, take the precipitate and add PBS buffer solution with pH 7.4, centrifuged for 15min to obtain the precipitate, continue the above operation 2-4 times, dissolve the precipitate with PBS buffer solution containing 2% BSA, filter to obtain the colloidal gold marker, add the colloidal gold marker to the micro Wells, freeze-dried to obtain gold-labeled microwell...

Embodiment 2

[0037] Example 2 The method of detecting ractopamine residues in raw milk and fresh milk with the colloidal gold kit of the above-mentioned β-stimulant drugs:

[0038] 1) Take 100 μL of milk sample and add it to the gold standard microwell, place it on a 50°C incubator and incubate for 5 minutes, and blow it with a small dropper to completely dissolve the red substance in the well;

[0039] 2) Insert the reagent strip into the gold-labeled microwell and incubate for 5 minutes;

[0040] 3) Take out the reagent strip from the microwell of the gold standard, and directly visually test the color intensity of the reagent strip, or interpret it with a matching colloidal gold reader;

[0041] 4) Interpretation of results

[0042] Such as image 3 Shown in a: Negative (-): The color of the T1 line is darker or the same as that of the C line, which is negative, indicating that the residual concentration of ractopamine in the sample is lower than the detection limit or there is no rac...

Embodiment 3

[0044] Example 3 Operation method for detecting salbutamol residues in raw milk and fresh milk with the colloidal gold kit of the above-mentioned β-agonist drugs

[0045] 1) Take 100 μL of milk sample and add it to the gold standard microwell, place it on a 50°C incubator and incubate for 5 minutes, and blow it with a small dropper to completely dissolve the red substance in the well;

[0046] 2) Insert the reagent strip into the gold-labeled microwell and incubate for 5 minutes;

[0047] 3) Take out the reagent strip from the microwell of the gold standard, and directly visually test the color intensity of the reagent strip, or interpret it with a matching colloidal gold reader;

[0048] 4) Interpretation of results

[0049] Such as image 3 Shown in middle b: Negative (-): The color of the T2 line is darker or as dark as that of the C line, indicating that the salbutamol drug residue concentration in the sample is lower than the detection limit or there is no salbutamol dr...

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Abstract

The invention discloses a colloidal gold kit for detecting a beta-stimulant drug. The kit comprises reagent strips and colloidal-gold marker micropores, the reagent strip comprises a base plate, a chromatography membrane, a sample pad arranged at two ends of the chromatography membrane, and an absorbent pad are arranged on the base plate in order, a quality control line C and a detection line arearranged on the chromatography membrane, the line C is coated with a goat-anti-mouse IgG antibody, the detection line comprises a T1 line and a T2 line, the T1 line is a ractopamine detection line, and is coated with a ractopamine antigen, the T2 line is the salbutamol, clenbuterol, brombuterol, tulobuterol, mabuterol, terbutaline, cimbuterol, clorprenaline, and cimaterol detection line, the T2 line is coated with a beta-stimulant antigen, the line C, the T1 line, and the T2 line present three parallel bands which are vertical to a long phase of a test paper strip, the colloidal-gold marker micropores contains a colloidal gold-labeled beta-stimulant polyclonal antibody and a ractopamine monoclonal antibody, by aiming at special objects of raw milk and fresh milk, ten drugs can be detectedat one time, a detection scope is enlarged, leak detection can be prevented, sensitivity is high, and stability is good.

Description

technical field [0001] The invention relates to a food safety rapid detection technology, in particular to a colloidal gold kit for detecting β-stimulant drugs and a preparation method thereof. Background technique [0002] With the improvement of people's living standards, the drug residues in food of animal origin have been paid more and more attention by the international community. β-stimulants are a class of chemically synthesized phenylethanolamine derivatives, which are commonly used clinically in the treatment of asthmatic diseases in humans and animals. When they are consumed continuously at doses higher than therapeutic levels, they can convert fat tissue in animals into muscle tissue. Improve the lean meat rate of animals and promote animal growth. Common beta-agonists are ractopamine, salbutamol, clenbuterol, bromobuterol, tobuterol, mabuterol, terbutaline, sibuterol, clorprenaline, sibuterol Luo and more than ten kinds. After long-term use of β-stimulants in ...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543G01N33/533G01N33/94
CPCG01N33/533G01N33/54306G01N33/558G01N33/9406
Inventor 王振国王伟萍罗南
Owner 杭州南开日新生物技术有限公司
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