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Preparation of ELISA kit for detecting plasticizer (DBP)

A plasticizer and kit technology, applied in the field of detection, can solve the problems of complex sample pretreatment methods, expensive instruments and equipment, and restrictions on wide application

Inactive Publication Date: 2014-07-02
DANYANG YITAI BIOTECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 of ELISA kit for detecting plasticizer (DBP)

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Embodiment Construction

[0025] ELISA kit for detecting plasticizer (DBP), which includes microtiter plate, plasticizer (DBP) standard, plasticizer (DBP) antibody working solution, enzyme-labeled secondary antibody working solution, substrate solution A, substrate Stock solution B, stop solution and concentrated wash solution.

[0026] Describe in detail below the preparation of the ELISA kit that detects plasticizer (DBP) among the present invention,

[0027] Preparation of plasticizer (DBP) protein conjugates:

[0028] Mix plasticizer (DBP) hapten and carrier protein BSA in 0.05 mol / L pH 9.6 carbonate buffer (CBS) at a ratio of 12:1, then add carbodiimide and stir for 1~ 2 h, react at room temperature for 24 h, and finally dialyze with 0.2 mol / L PBS pH 7.6 for two days to remove unreacted hapten to obtain plasticizer (DBP) protein conjugate.

[0029] Preparation of plasticizer (DBP) antibody:

[0030] 3-month-old healthy white rabbits were selected and passed through three immunizations. The dosa...

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Abstract

The invention relates to preparation of an ELISA kit for detecting plasticizer (DBP). The kit with sensitive detection, accuracy, fastness, simple operation and strong specificity is applicable to determination of a large number of samples. The kit includes an ELISA plate coated with plasticizer (DBP) antigens, a plasticizer (DBP) standard sample, a plasticizer (DBP) antibody working solution, an enzyme-labeled second antibody working solution, a substrate solution A, a substrate solution B, a stop solution, a concentrating sample weak solution and a washing concentrate. The principle of the plasticizer (DBP) detection kit is solid phase indirect competitive enzyme-linked immunosorbent assay. An extracted sample, the enzyme-labeled standard second antibody working solution and the antibody working solution are added to corresponding enzyme-labeled holes; after incubation for a period of time, the substrate solution A and the substrate solution B are added into a washing plate; under the action of the enzyme, blue color occurs in the holes; and the stop solution is added in, and the color changes from blue to yellow. The level of coloration is in an inverse proportion relation with the content of plasticizer (DBP) in the standard substance or the sample. The method can be directly applied to detection of plasticizer (DBP) content in liquor samples.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to an enzyme-linked immunosorbent assay kit and a detection method for quantitatively detecting residual plasticizer (DBP) in liquor samples. Background technique [0002] A plasticizer is an additive that increases the softness or liquefaction of a material. Phthalate ester plasticizers are classified as suspected environmental hormones, and their biological toxicity is mainly estrogenic and antiandrogenic, which can cause endocrine disorders and damage the reproductive function of organisms, including reduced reproductive rate, miscarriage, and birth defects , Abnormal sperm count, testicular damage, can also cause malignant tumors, cause deformed children, and lead to precocious puberty in children. [0003] Dibutyl phthalate (Dibutyl phthalate, DBP) is a commonly used plasticizer, also used as an additive in adhesives and printing inks, and can also be used as an ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/535
CPCG01N33/543
Inventor 杜道林曾昆洪霞杜霞
Owner DANYANG YITAI BIOTECH DEV
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