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Five-conjugate testing card for detecting beta-stimulant drugs and preparation method thereof

A stimulant and testing card technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of expensive equipment, complicated operation, difficult to popularize, etc., and achieve the effect of simple method and accurate results.

Inactive Publication Date: 2013-01-30
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the detection of common β-agonists clenbuterol, ractopamine, albuterol, phenylethanolamine A and terbutaline mainly includes enzyme-linked immunosorbent immunoassay (ELISA), gas chromatography-mass spectrometry (GC-MC), high-performance liquid Detection methods such as phase chromatography (HPLC), but these methods require expensive equipment, complicated operations, and are difficult to popularize

Method used

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  • Five-conjugate testing card for detecting beta-stimulant drugs and preparation method thereof
  • Five-conjugate testing card for detecting beta-stimulant drugs and preparation method thereof
  • Five-conjugate testing card for detecting beta-stimulant drugs and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0023] figure 1 It is the structural diagram of the five-link colloidal gold detection card of the present invention, figure 2 It is the cross-sectional structural diagram of the test strip in the five-link colloidal gold detection card of the present invention: among the figure, 13 is a PVC rubber sheet; 11 is a sample pad; 12 is a colloidal gold binding pad, and the colloidal gold binding pad is coated with monoclonal Antibody colloidal gold marker; 14 is a coating membrane, namely a nitrocellulose membrane, which is coated with clenbuterol-BSA, ractopamine-BSA, salbutamol-BSA, phenylethanolamine A-BSA, terbu Taline-BSA and rabbit anti-mouse IgG; 15 for absorbent pads.

[0024] The sample pad 11 and the colloidal gold bonding pad 12 are adhered to one end of the PVC rubber plate 13 (the sample loading end), and the sample pad 11 is lapped on the colloidal gold bonding pad 12 .

[0025] A nitrocellulose membrane 14 is adhered in the middle of the PVC rubber sheet 13 . On ...

Embodiment 2

[0028] The preparation of clenbuterol, ractopamine, salbutamol, phenylethanolamine A and terbutaline five-unit colloidal gold detection card is realized by the following steps:

[0029]1. Preparation of colloidal gold solution: Take a 1 L Erlenmeyer flask, add 495 mL of ultrapure water, and then add 1% chloroauric acid (HAuCl 4 ·3H 2 O) 5 mL, prepared into 500 mL 0.01% chloroauric acid aqueous solution, heated to boil, then added 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) Solution 5-7 mL, continue to stir and heat, when the color of the solution turns completely transparent purple red, stop heating after maintaining for 5 minutes, add water to the original volume, cool to room temperature, and store at 2-8°C for later use;

[0030] 2. Antibody pretreatment: centrifuge clenbuterol, ractopamine, salbutamol, phenylethanolamine A and terbutaline monoclonal antibodies at 1000 r / min, 4°C for 20 min, and take the supernatant. Dilute with 0.01 mol / L PBS to 1 mg / mL; or use 0....

Embodiment 3

[0037] Minimum detection limit test

[0038] Prepare standard clenbuterol, ractopamine, albuterol, phenylethanolamine A and terbutaline standard solution concentrations: 0, 0.5 ng / mL, 1 ng / mL, 2.5 ng / mL, 5 ng / mL; Add 2 drops of the sample to the sample hole of the detection card, and observe the color development results of the detection line T and the quality control line C after 5 minutes. After multiple tests, the detection rates of clenbuterol, ractopamine, salbutamol, phenylethanolamine A and terbutaline were over 99%, and the minimum detection limit was less than 5 ng / mL.

[0039]

[0040] .

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Abstract

The invention relates to a five-conjugate testing card for detecting beta-stimulant drugs and a preparation method thereof and belongs to the technical field of beta-adrenergic receptor stimulant detection. A test strip is arranged in a casing of the testing card which is composed of a polyvinyl chloride (PVC) glue board, a sample pad, a colloidal gold combination pad, a coated film and absorbent cotton. A colloidal gold film is a glass cellulose film which contains colloidal gold markers of a clenbuterol monoclonal antibody, a ractopamine monoclonal antibody, a salbutamol monoclonal antibody, a phenylethanolamine A monoclonal antibody and a terbutaline monoclonal antibody. The coated film is a nitrocellulose film, wherein five detection lines T and a quality control line C are arranged on the coated film, and the quality control line C is coated with a rabbit anti-mouse immunoglobulin G (IgG) antibody. The five-conjugate testing card can simultaneously detect out clenbuterol, ractopamine, salbutamol, phenylethanolamine A and terbutaline, and is simple, convenient and fast in method, and accurate in result.

Description

technical field [0001] The invention relates to the technical field of detection of β-adrenergic receptor agonists, in particular to a five-unit detection card for clenbuterol, ractopamine, albuterol, phenylethanolamine A and terbutaline and a preparation method thereof. Background technique [0002] With the development of scale and commercialization of animal husbandry, veterinary drugs and feed additives have been widely used in animal husbandry production. Veterinary drugs are widely used to effectively control various diseases and reduce animal mortality; Increased animal weight gain rate and shortened animal feeding cycle, improved leather quality, reduced fasting carcass and animal tissue waste rate; promoted the growth of animal-derived product production and the development of intensive animal farming. Current veterinary drugs are not only used to prevent and treat animal diseases, but more and more drugs are used for non-therapeutic purposes such as promoting growt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 杜道林洪霞薛永来张祯
Owner JIANGSU WISE SCI & TECH DEV
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