Clenobuterol hydrochloride detecting test paper and detecting method thereof

A technology of clenbuterol hydrochloride and detection test paper, which is applied in the field of clenbuterol hydrochloride detection test paper and its detection, can solve the problems of expensive equipment, false negative detection line, long time, etc., so that false negative and false positive are not easy Effect of misjudgment, high specificity and sensitivity, and image of test results

Inactive Publication Date: 2009-06-17
周炬华 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the cumbersome test operation and long time of the above-mentioned methods, professionally trained and experienced operators are required, the required instruments and equipment are relatively expensive, and the detection cost is high. It is not suitable for large-scale rapid detection in the field or in the field.
Enzyme-linked immunosorbent assay (ELISA) is also a commonly used detection technology. ELISA can detect multiple samples, as well as qualitative and quantitative detection. However, there is no commercial kit for this product on the market, and ELISA requires certain operations. For experienced professionals, the operation process is relatively complicated, and the detection time is relatively long. Different personnel often have different detection results, and a matching microplate reading instrument is required, so it is difficult to implement on-site detection.
[0004] The existing clenbuterol hydrochloride immunochromatography test paper adopts the immunochromatographic competition method, and in the detection process, clenbuterol hydrochloride and the solid-phased clenbuterol hydrochloride conjugate at the detection line compete for the colloidal gold-labeled single Cloned antibody, when the sample contains more than 5ng / ml of clenbuterol hydrochloride, the detection line does not develop color, and when the content of clenbuterol hydrochloride is less than 5ng / ml, the detection line develops color, the content of clenbuterol hydrochloride in the sample It is negatively correlated with the color development of the detection line, and people's judgment thinking is first of all positive correlation, so it is very easy to make interpretation errors, and in the case of negative correlation, it often occurs whether the detection line has color development and unnecessary false negatives cause omissions. check

Method used

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  • Clenobuterol hydrochloride detecting test paper and detecting method thereof
  • Clenobuterol hydrochloride detecting test paper and detecting method thereof
  • Clenobuterol hydrochloride detecting test paper and detecting method thereof

Examples

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Embodiment 1

[0031] Example 1: Preparation of colloidal gold detection test paper for clenbuterol hydrochloride

[0032] 1. Preparation of clenbuterol hydrochloride-coupled carrier protein solution interception blotting solution

[0033] (1) Using diazo coupling reaction, with 500 μL of 0.25mol / L H 2 SO 4 Dissolve 10mg clenbuterol hydrochloride, take 200μL 2% NaNO 2 The solution was slowly dropped into the above solution, and reacted at 4°C for 10 minutes.

[0034] (2) Under magnetic stirring conditions, drop the resulting mixed solution into 2 mL of bovine serum albumin (BSA), chicken ovalbumin (OVA), ferritin (FE) and hemocyanin (KLH) solutions respectively in advance, Adjust the pH value to 9.5 with 1mol / L NaOH, and slowly stir overnight at 4°C to form four linked products linked by azo bonds.

[0035](3) The obtained four connection products were put into dialysis bags respectively, dialyzed with 10mM pH7.4 phosphate buffer solution for three days, and changed twice a day to remove...

Embodiment 2

[0045] Example 2: Preparation of clenbuterol hydrochloride nanolatex-labeled antibody solution and nanolatex-labeled IgG solution glass wool

[0046] After the nano-latex particles are activated and the antibody or IgG to be labeled is mixed in proportion, adjust the pH value to the required range, add a coupling agent (such as EDC, NHS, etc.) and wait until the reaction is complete. Add 10% BSA to a final concentration of 1%, and mix thoroughly. Let stand evenly for 10 minutes, centrifuge at 12000rpm at 4°C for 25 minutes, discard the supernatant to remove antibodies not bound to the nanolatex particles, and use pH8.2, 20mM Tris-HCL buffer protection solution (preparation: Tris3.0285g, HCL0.35ml, BSA10g , sucrose 50g, NaN 3 2g, PEG2000 2g, deionized water 900ml, adjust the pH to 8.2 with 1Mol NaOH, add deionized water to 1000mL) and resuspend to obtain the nanolatex-labeled antibody solution. Spread the nano-emulsion-labeled antibody solution on the fiber glass wool, and ev...

Embodiment 3

[0047] Example 3: Preparation of clenbuterol colloidal selenium hydrochloride, colloidal iron-labeled antibody solution and colloidal selenium, colloidal iron-labeled IgG solution glass wool

[0048] Use 0.5mol / L K for colloidal selenium and colloidal iron 2 CO 3 When the pH value of the gold-adjusting gum solution is about 8.2, add the required amount of clenbuterol hydrochloride antibody. After 10 minutes, add 10% BSA to a final concentration of 1%. Clear to remove the antibody that is not combined with colloidal selenium and colloidal iron, pH8.2, 20mM Tris-HCL buffer protection solution (preparation: Tris3.0285g, HCL0.35ml, BSA10g, sucrose 50g, NaN 3 2g, 2g of PEG2000, 900ml of deionized water, adjust the pH to 8.2 with 1Mol NaOH, then add deionized water to 1000mL) for resuspension, and obtain the antibody solution labeled with colloidal selenium and colloidal iron. Spread the colloidal selenium and colloidal iron-labeled antibody solution on fiberglass wool, and evenly...

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Abstract

The invention discloses a Clenbuterol hydrochloride detection test paper and a detection method thereof. The Clenbuterol hydrochloride detection test paper comprises a support and fixing lining which is sequentially provided with a sample adsorption layer, a combination pad, a microporous siphon reaction layer and a water absorption layer; the combination pad is provided with a Clenbuterol hydrochloride antibody labeled by a trace particle and an antibody A labeled by the trace particle, and the antibody A is different from the Clenbuterol hydrochloride antibody; the microporous siphon reaction layer is sequentially provided with a blotting area and a detection area of Clenbuterol hydrochloride carrier protein solution, and the detection area comprises a detection solution blotting area and a control solution blotting area. Two lines occur in the detection area when the content of the Clenbuterol hydrochloride is more than 5 nanogram in a sample; and one line occurs in the detection area when the content of the Clenbuterol hydrochloride is less than 5 nanogram in the sample. The test paper has a detection mode and a reading manner equivalent to those of a sandwich method and low cost. The detection method is simple and rapid, and has simple operation.

Description

technical field [0001] The invention relates to the technical field of immunological analysis of veterinary drug residues, in particular to a clenbuterol hydrochloride detection test paper and a detection method thereof. Background technique [0002] Clenbuterol hydrochloride (clenbuterol, CLB), also known as clenbuterol, ammonia clenbuterol, clenbuterol, is a β2-adrenergic receptor agonist (abbreviated as β-stimulant). The chemical name is hydroxymethadrenaline, white or off-white crystalline powder, odorless, bitter taste, melting point 161°C. The drug can selectively act on adrenergic receptors and is a powerful agonist that can cause sympathetic nerve excitement. Animals eat feed containing clenbuterol hydrochloride, which can improve the metabolic pathway of nutrients and promote animal muscle. In particular, the synthesis of bone protein can inhibit the accumulation of fat synthesis, thereby accelerating the growth rate of animals, and the lean meat is relatively incr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
Inventor 周炬华李绍旭
Owner 周炬华
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