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Kit for detecting clenobuterol hydrochloride residue and method for preparing same

A technology of clenbuterol hydrochloride and a kit is applied to a kit for detecting clenbuterol in pork and its preparation, and the field of the kit can solve the problems of complicated operation, difficulty in meeting detection requirements by color ELISA, expensive equipment and the like , to meet the detection requirements, improve stability, and improve the effect of sensitivity

Inactive Publication Date: 2008-11-19
HARBIN RENHUANG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both HPLC-MS and GC-MS methods require expensive equipment and complex operations, and are usually used as large-scale screening samples
However, the detection limit of the commonly used chromogenic ELISA is generally around 0.1ug / L, and the maximum allowable residue of CLB is also 0.1ug / L. Therefore, it is difficult for the chromogenic ELISA method to meet the detection requirements.
At present, there is no development and production of pork fine quantitative chemiluminescence detection kits at home and abroad. The main technical difficulties are the mastery and application of new chemiluminescence technologies, how to improve sensitivity and specificity, etc.

Method used

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  • Kit for detecting clenobuterol hydrochloride residue and method for preparing same
  • Kit for detecting clenobuterol hydrochloride residue and method for preparing same

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Embodiment Construction

[0023] 1. Preparation of hybridoma cell line secreting anti-clenbuterol monoclonal antibody and enzyme-labeled monoclonal antibody

[0024] 1. Preparation of Complete Antigen

[0025] 1.1 Test material

[0026] 1.1.1 Clenbuterol hydrochloride (purchased from Sigma)

[0027] 1.1.2 Bovine Serum Albumin (BSA)

[0028] 1.1.3 Cold hydrochloric acid

[0029] 1.1.4NaNO 3

[0030] 1.2 Preparation process

[0031] Take 660ul of 1.0mg / ml clenbuterol hydrochloride solution, adjust the pH to 2.5 with cold hydrochloric acid, add NaNO 3 , to azotize clenbuterol hydrochloride, add the azotized clenbuterol hydrochloride to BSA and OVA solution, put it in the refrigerator at 4°C overnight, and take the dialysis the next day, all the above operations are in a dark and dark environment at 4°C After the dialysis, measure the concentration, dilute to 1.0 mg / ml, subpackage, and freeze at 20°C for later use, wherein CLB-BSA is coated with antigen, and CLB-OVA is used for immunizing antigen. ...

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Abstract

The invention discloses a hybridoma cell line secreting a monoclonal antibody repelling Clenbuterol hydrochloric acid, and an antibody repelling Clenbuterol hydrochloric acid through the hybridoma cell line. The invention also discloses a kit for detecting Clenbuterol hydrochloric acid residual. The kit comprises: a glowing black test board coated by CL-BSA in advance, a Clenbuterol hydrochloric acid standard liquid, an ELISA Clenbuterol hydrochloric acid antibody, a chemical luminescence liquid and a cleaning solution. The kid for detecting Clenbuterol hydrochloric acid residual has qualitative improvements on specificity, sensitivity and stability compared with the prior kit. The lowest detection value of Clenbuterol (CLB) hydrochloric acid residual is 0.02 ng / L, thereby completely meeting the requirements of detecting the level CLB residual.

Description

technical field [0001] The invention relates to a kit, in particular to a kit for detecting clenbuterol in pork and a preparation method thereof, belonging to the field of biotechnology. Background technique [0002] Clenbuterol (CLB), commonly known as "pork essence", has toxic and side effects on human liver, kidney and nerves. At present, the methods for detecting CLB mainly include high-performance liquid chromatography-mass spectrometry (HPLC-MS), gas chromatography-mass spectrometry (GC-MS) and ELISA methods. Some people have studied the detection of clenbuterol hydrochloride residues in meat samples by solid phase extraction GC-MS and the analysis of clenbuterol enantiomers by HPLC chiral stationary phase. Both HPLC-MS and GC-MS methods require expensive equipment and complex operations, and are usually used as large-scale screening samples. However, the detection limit of the commonly used chromogenic ELISA is generally around 0.1ug / L, and the maximum allowable res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/18C07K16/18G01N33/577
Inventor 李绍铭
Owner HARBIN RENHUANG PHARMA
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