Colloidal gold immune test paper for rapid detection of dibutyl phthalate as well as preparation method and application thereof

A technology of dibutyl phthalate and colloidal gold, which is applied in the field of rapid detection of residual immunochemistry, can solve the problems of fast and simple detection of agricultural products, not suitable for rapid detection on site, and small amount of samples for one detection. The effect of fast speed, short detection time and low cost

Inactive Publication Date: 2012-09-19
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these methods require complex pretreatment, expensive equipment, time-consuming, laborious and costly detection. Compared with other detection systems, the amount of samples for one detection is small and the detection time is long, which is not suitable for rapid on-site detection and is only used by a small number of laboratories. Confirmation methods cannot meet the needs of quick and easy detection of the environment and agricultural products

Method used

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  • Colloidal gold immune test paper for rapid detection of dibutyl phthalate as well as preparation method and application thereof
  • Colloidal gold immune test paper for rapid detection of dibutyl phthalate as well as preparation method and application thereof
  • Colloidal gold immune test paper for rapid detection of dibutyl phthalate as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The colloidal gold immunoassay paper for rapid detection of dibutyl phthalate of the present invention comprises a reaction membrane 2, a sample pad 3, a conjugate release pad 4, a water-absorbing pad 5 and a backing 1, and is wrapped on the reaction membrane 2. Test area 6 with dibutyl phthalate-carrier protein conjugate and quality control area 7 with goat anti-mouse IgG, conjugate release pad 4 coated with dibutyl phthalate monoclonal antibody-colloid Gold marker 8.

[0038] In this embodiment, the sample pad 3 , the conjugate release pad 4 , the reaction membrane 2 and the water-absorbing pad 5 are sequentially arranged on the backing 1 .

[0039] Dibutyl phthalate-carrier protein conjugate is obtained by nitrating and reducing dibutyl phthalate to form 4-aminodibutyl phthalate hapten, and then coupling with carrier protein. Wherein, the carrier protein in the dibutyl phthalate-carrier protein conjugate is bovine serum albumin. Dibutyl phthalate monoclonal antibod...

Embodiment 2

[0041] Colloidal gold immunoassay paper preparation method for rapid detection of dibutyl phthalate of the present invention comprises the following steps:

[0042] 1. Synthesis and identification of dibutyl phthalate-carrier protein conjugates

[0043] (1) Synthesis of 4-aminophthalate-dibutyl-bovine serum albumin

[0044] Dissolve 100mg of carbodiimide in 3.0ml of 10mol / L PBS solution with pH 8.0 (referred to as 1 solution); take 10mg of 4-aminodibutylphthalate and dissolve it in 1ml of dimethylformamide (2 solutions) ; Dissolve 20 mg of bovine serum albumin in 1ml of 10mol / L PBs (pH8.0) solution (3 solutions); mix (2 solutions) with (3 solutions), and gradually add (1 solution) under magnetic stirring, Stir at 4°C for 12h. Distilled water was used to fully dialyze it for 4-6 days to obtain the immune antigen.

[0045] 2. Preparation of dibutyl phthalate monoclonal antibody

[0046] (1) Animal immunity

[0047] The immunized antigen was injected into Balb / c mice at an i...

Embodiment 3

[0066] Example 3: Detection of residues in samples

[0067] 1. Sample pretreatment

[0068] Weigh 50g of chicken, pork, pork liver, sausage, fish, shrimp, honey, puffed food, etc., homogenate, put into a stoppered conical flask, add 100ml of extractant (methanol, ethyl acetate, etc.), extract for 1h . Centrifuge, take the supernatant, concentrate to 1ml, and dilute with PBS or normal saline.

[0069] 2. Detection with test paper card

[0070] Drop the sample into the hole of the reagent card with a dropper, add 3 drops, count the time after adding, and observe the result within 5-8 minutes. The detection and interpretation of samples exceeding 10 minutes is invalid.

[0071] 3. Analysis of test results

[0072] Positive: When the (C) area shows a red band, but the (T) area does not show color, it is judged as positive.

[0073] Negative: When (C) area shows a red band, (T) area also shows a red band, and the color of (T) area is close to or lighter than that of (C) area,...

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Abstract

The invention relates to a colloidal gold immune test paper for rapid detection of dibutyl phthalate as well as a preparation method and application thereof. The colloidal gold immune test paper comprises a reaction film, a sample pad, a conjugate releasing pad, an absorbent pad and a back pad. The reaction film is coated with a testing area of a dibutyl phthalate-carrier protein conjugate and a quality control area of goat-anti-mouse IgG; the conjugate releasing pad is coated with a dibutyl phthalate monoclonal antibody, namely a colloid metal marker. The test paper provided by the present invention can be used for detecting the residual quantity of the dibutyl phthalate drugs in the animal-derived food, such as the chicken, the pork, the urine, and the honey. The test paper has the advantages of simple operation, low cost, high sensitivity, and quick test speed, is applicable for large-scale popularization and mass sample screening, can be controlled in the scene, does not need special instruments and specialized persons, and is easy to implement.

Description

technical field [0001] The invention relates to a colloidal gold immunological test paper for rapid detection of residual dibutyl phthalate by means of an immunochromatographic reaction of colloidal gold labeling and color development, which belongs to the immunochemical rapid detection of residual dibutyl phthalate technology field. Background technique [0002] Phthalates (PAEs), also known as phthalates, are a type of environmental estrogen and one of the commonly used plasticizers. It is widely used in hundreds of products such as toys, food packaging materials, medical blood bags and hoses, vinyl floors and wallpapers, cleaners, lubricants, personal care products (such as nail polish, hair sprays, soaps and shampoos) middle. Phthalates can interfere with endocrine, reduce the number of male sperm, reduce motility, and abnormal shape. In severe cases, it can also cause dead sperm and testicular cancer. It is the "culprit" of male reproductive problems and even affects ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/531
Inventor 孔涛杨自军张才赵振升郝雪琴周变华常景周李润乐魏迎军李嘉嘉王振华
Owner HENAN UNIV OF SCI & TECH
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