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Preparation method of enzyme-linked immunosorbent assay (ELISA) kit for detecting cyproheptadine hydrochloride

A technology of cyproheptadinase hydrochloride and cyproheptadine hydrochloride, applied in the field of detection, can solve the problems of high detection limit, expensive instruments and equipment, complex sample pretreatment methods, etc., and achieve the effect of sensitive detection and easy operation

Inactive Publication Date: 2016-02-03
JIANGSU WISE SCI & TECH DEV
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Problems solved by technology

Among them, high performance liquid chromatography (HPLC) and gas chromatography (GC) are quantitative detection methods, and the results are accurate and reliable. The disadvantages are that the detection limit is high, the equipment is relatively expensive, and the complicated sample pretreatment method limits the wide application of the method. application

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  • Preparation method of enzyme-linked immunosorbent assay (ELISA) kit for detecting cyproheptadine hydrochloride

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Embodiment Construction

[0024] An ELISA kit for detecting cyproheptadine hydrochloride, which includes an enzyme plate, cyproheptadine hydrochloride standard substance, cyproheptadine hydrochloride enzyme-labeled antibody working solution, substrate solution A, substrate solution B, stop solution and concentrated washing solution.

[0025] Describe in detail below the preparation of the ELISA kit that detects cyproheptadine hydrochloride among the present invention,

[0026] Preparation of Cyproheptadine Hydrochloride Protein Conjugate:

[0027] Mix the cyproheptadine hydrochloride hapten and the carrier protein BSA in a 0.05mol / L pH9.6 carbonate buffer (CBS) at a ratio of 12:1, then add carbodiimide, and stir for 1-2 hours. React at room temperature for 24 hours, and finally dialyze with 0.2mol / L PBS at pH 7.6 for two days to remove unreacted hapten to obtain cyproheptadine hydrochloride protein conjugate.

[0028] Preparation of cyproheptadine hydrochloride antibody:

[0029] 3-month-old healthy ...

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Abstract

The invention provides a preparation method of enzyme-linked immunosorbent assay (ELISA) kit for detecting cyproheptadine hydrochloride. The kit is sensitive, accurate and rapid in detection, simple and convenient to operate, high in specificity and suitable for detecting samples in large batch, and comprises an ELISA plate coated with a cyproheptadine hydrochloride antigen, a cyproheptadine hydrochloride standard substance, a cyproheptadine hydrochloride enzyme-labeled antibody working solution, a substrate solution A, a substrate solution B, a stop solution and a concentrated scrubbing solution. According the principle of the kit, an ELISA reaction is performed through direct solid-phase competition; an extracted sample and the enzyme-labeled antibody working solution are added into a corresponding ELISA hole; after a period of incubation, the ELISA plate is scrubbed, and the substrate solution A and the substrate solution B are added; under the enzymatic action, a blue color appears in the ELISA hole; after the stop solution is added, the color changes from blue to yellow, and the color depth has an inversely proportional relationship with the content of cyproheptadine hydrochloride in the standard substance or the sample. The principle can be directly utilized for detecting the content of cyproheptadine hydrochloride in a swine urine sample.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to an enzyme-linked immunoimmunoassay kit and a detection method for quantitative detection of cyproheptadine hydrochloride residues in pig urine samples. Background technique [0002] Cyproheptadine hydrochloride is an H1 receptor antagonist, belonging to antihistamines, but it is also a 5-hydroxytryptamine receptor antagonist. It is a legal human drug to resist the effect of histamine on blood vessels and bronchial smooth muscle in the body, thereby eliminating allergies Symptoms, it can also be used to improve the patient's appetite. The use of cyproheptadine hydrochloride in the breeding industry can promote animal growth. It has the effect of anti-serotonin, which can inhibit the satiety center of the hypothalamus and stimulate appetite to increase weight. Therefore, some animal husbandry enterprises use it as an illegal feed additive for Promote the growth of li...

Claims

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Application Information

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IPC IPC(8): G01N33/577
Inventor 洪霞薛永来
Owner JIANGSU WISE SCI & TECH DEV
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