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Ractopamine, salbutamol, cimaterol triple colloidal gold test strip and preparation method and use thereof

A technology of ractopamine and colloidal gold test paper, which is applied in the detection of β2-receptor agonist residues and food safety, and can solve problems such as large background interference, limited detection sensitivity, and obvious matrix effects of samples

Active Publication Date: 2015-12-09
江西中德生物工程股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] (1) Only when the gold particles gather to a certain amount (10 7 piece / mm 2 ), the purple-red band visible to the naked eye appears, and the contrast between the band and the background is not large, thus limiting the detection sensitivity
[0006] (2) The sample matrix effect is obvious and the background interference is large
[0007] (3) Quantitative detection cannot be realized
[0008] (4) Only a single pollutant can be detected at a time
[0009] In addition, although there are now test cards that can individually detect ractopamine, albuterol, and cimaterol, so far, there is still no product that can simultaneously qualitatively and quantitatively detect ractopamine, albuterol, and cimaterol.
[0010] Therefore, the current detection methods for the simultaneous quantitative detection of ractopamine, albuterol and cimaterol still need to be improved.

Method used

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  • Ractopamine, salbutamol, cimaterol triple colloidal gold test strip and preparation method and use thereof
  • Ractopamine, salbutamol, cimaterol triple colloidal gold test strip and preparation method and use thereof
  • Ractopamine, salbutamol, cimaterol triple colloidal gold test strip and preparation method and use thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0040] Synthesis of embodiment 1 immune antigen

[0041] 1.1 Synthesis of ractopamine immune antigen

[0042] Prepare ractopamine-BSA immunoantigen by mixed acid anhydride method: Weigh 34 mg of ractopamine and 10 mg of succinic anhydride to react in 2 mL of pyridine, stir overnight at room temperature, and place pyridine in a fume hood to completely evaporate. The reaction product is ractopamine-BSA Hemisuccinic anhydride; dissolve it in 2 mL of N,N-dimethylformamide and 2 mL of 1,4-dioxane mixture, add 26.2 μL of tri-n-butylamine, stir in an ice bath for 10 minutes, add 15L of isobutyl chloroformate was reacted at room temperature and stirred for 1 hour; this mixture was added dropwise to a pre-cooled protein solution (100mgBSA dissolved in 0.1M sodium borate pH8.5), reacted at room temperature overnight, and dialyzed in PBS for more than 72 hours. After dialysis, the purified ractopamine-BSA immune antigen was obtained.

[0043]

[0044] 1.2 Synthesis of albuterol immune...

Embodiment 2

[0050] Example 2 Preparation and Titer Detection of Immunogen Monoclonal Antibody

[0051] 2.1 Preparation of ractopamine monoclonal antibody

[0052] Take 8-week-old female BALB / c mice, and use 0.1mL ractopamine-BSA and an equal volume of complete Freund's adjuvant emulsion to immunize each mouse for the first time by intraperitoneal injection; after that, take the same dose of immunogen to add With incomplete Freund's adjuvant, the same method was used to boost immunization once every month; the antibody titer was measured by indirect ELISA 10 to 14 days after the third and fourth immunizations, and finally the one with the highest antibody titer was selected for cell fusion. Spleen cells from immunized mice were taken under sterile conditions, fused with SP2 / O myeloma cells at a ratio of 5:1, added to HAT medium, 37 degrees Celsius, 6% CO 2 Cultivate in an incubator; replace half of the medium with fresh HAT medium after 5 days, and replace the HAT medium with HT medium af...

Embodiment 3

[0065] Embodiment 3 Preparation of ractopamine, salbutamol, cimaterol triple colloidal gold test strip

[0066] 3.1 Preparation of colloidal gold

[0067] The basic principle of immune colloidal gold preparation is that chloroauric acid polymerizes into gold particles of a certain size under the action of a reducing agent to form a negatively charged hydrophobic colloid solution that is stable due to electrostatic interaction. The present invention adopts trisodium citrate reduction method to prepare colloidal gold, and specific process is as follows: take 0.01% chloroauric acid 100mL aqueous solution and heat to boiling, accurately add 1.5mL of 1% trisodium citrate aqueous solution under agitation, golden chloroauric acid is in Turn into purple within 2 minutes, turn off the heat source, continue to stir at high speed for 10 minutes, then reduce the speed to low gear, continue to stir for 1 hour, and restore the original volume with distilled water after cooling, which is the p...

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Abstract

The invention brings forward a colloidal gold triple-test strip for ractopamine, salbutamol and cimaterol and a preparation method thereof. The colloidal gold triple-test strip for ractopamine, salbutamol and cimaterol comprises a base plate; the base plate has a first end and a second end and is successively provided with, from first end and second end, a filter paper, a sample pad, a colloidal gold pad, a cellulose nitrate membrane and a water absorbing pad, wherein the colloidal gold pad contains a colloidal gold-labeled monoclonal antibody to ractopamine, a colloidal gold-labeled monoclonal antibody to salbutamol and a colloidal gold-labeled monoclonal antibody to cimaterol, and three detection lines and one quality control line are further formed on the cellulose nitrate membrane. Cooperation between the test strip and a colloidal gold reading instrument enables quantitative determination of ractopamine, salbutamol and cimaterol to be effectively realized.

Description

technical field [0001] The invention belongs to the field of food safety and involves β 2 - Field of detection of receptor agonist residues. Specifically, the present invention relates to ractopamine, salbutamol, cimaterol dual colloidal gold test strips and a preparation method and use thereof. Background technique [0002] Ractopamine, Salbutamol, and Cimaterol are all beta 2 -receptor agonists, and in recent years they have been used as the main substitute for clenbuterol. It is well known that β 2 - Receptor agonist drugs used in animal husbandry can improve feed conversion rate and increase lean meat production. beta 2 - Receptor agonists can cause heart palpitations, headaches, dizziness, nausea, and even liver and kidney damage. my country and many other countries in the world have listed this type of drug as a banned drug. However, driven by economic interests and lack of an effective detection system, the current breeding Realms still exist beta 2 -Illegal use...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/577
Inventor 赖卫华彭涛杨万春陈媛刘文娟
Owner 江西中德生物工程股份有限公司
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