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Method and enzyme-linked immunosorbent assay (ELISA) kit for detecting sunset yellow

A technology of enzyme-linked immunosorbent reagents and sunset yellow, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effects of high accuracy, simplified steps, and guaranteed reliability

Inactive Publication Date: 2012-01-11
JIANGSU BERKGEN BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, my country's scientific research workers have done a lot of work on the detection of sunset yellow, but they all mainly focus on the physical and chemical detection. The literature search has not yet found the report about the EL ISA detection method of sunset yellow. In view of this, the present invention researches and establishes A Competitive EL ISA Assay for Sunset Yellow

Method used

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  • Method and enzyme-linked immunosorbent assay (ELISA) kit for detecting sunset yellow
  • Method and enzyme-linked immunosorbent assay (ELISA) kit for detecting sunset yellow
  • Method and enzyme-linked immunosorbent assay (ELISA) kit for detecting sunset yellow

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1. A kit and ELISA detection method using a hapten-carrier protein conjugate as a coating source and an enzyme-labeled antibody as an enzyme marker

[0043] 1. ELISA detection principle using sunset yellow hapten and carrier protein conjugate as coating source and enzyme-labeled antibody as enzyme marker:

[0044] When the coating on the microwell strip of the microtiter plate was originally a sunset yellow hapten-carrier protein conjugate, after adding the standard solution or sample solution to the microwell of the enzyme plate, the sunset yellow in the sample and the microtiter plate On the sunset yellow-coupled antigen competition enzyme-labeled sunset yellow-specific antibody, wash the plate, develop color, the absorbance value of the sample is negatively correlated with the content of sunset yellow, and the content of sunset yellow in the sample can be obtained by comparing with the standard curve. At the same time, the content of sunset yellow in the sample...

Embodiment 2

[0103] Example 2. Kit and ELISA detection method using hapten-carrier protein conjugate as coating source and enzyme-labeled secondary antibody as enzyme label

[0104] 1. ELISA detection principle using sunset yellow hapten-carrier protein conjugate as coating source and enzyme-labeled secondary antibody as enzyme marker:

[0105] When the coating on the microwell strip of the microtiter plate is originally a sunset yellow hapten-carrier protein conjugate, after adding the standard solution or sample solution to the microwell of the microtiter plate, add the sunset yellow specific antibody, and the sample The sunset yellow in the medium competes with the sunset yellow-coupled antigen on the ELISA plate for the sunset yellow-specific antibody, washes the plate, then adds the enzyme-labeled anti-antibody for amplification, and develops the color with a chromogenic solution. The absorbance value of the sample is negatively correlated with the content of the sunset yellow , compa...

Embodiment 3

[0115] Embodiment 3, test kit precision, accuracy, preservation test

[0116] 1. The precision test of the kit

[0117] (1) Repeatability test of standard solution

[0118] From 3 batches of enzyme-labeled plates prepared according to the method in Example 1, 10 microwells were taken out respectively, and the absorbance value (OD value) of the 9 μg / mL standard solution was measured, repeated 10 times, and the coefficient of variation CV was calculated. The results are shown in Table 1.

[0119] Table 1 Standard solution repeatability test

[0120] cv%

1

2

3

4

5

6

7

8

9

10

01 batch

8

6.5

7.3

6.6

6.9

7.1

8.2

6.8

7.4

8.1

02 batches

8.2

6.8

6.7

6.9

7.2

7.4

8.4

6.3

6.6

8.3

03 batches

7.2

6.8

6.3

6.1

8

7.5

6.3

7.6

8.1

...

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Abstract

The invention discloses a method and enzyme-linked immunosorbent assay (ELISA) kit for detecting sunset yellow. The ELISA kit for detecting sunset yellow comprises sunset yellow hapten and a specific antibody of sunset yellow, wherein the specific antibody is a polyclonal antibody or monoclonal antibody of sunset yellow. A high-specificity monoclonal antibody of sunset yellow is adopted in the kit, thus ensuring the reliability of the detection result. The experimental results show that the kit has the characteristics of high specificity, high sensitivity, high precision, high accuracy and the like. Main reagents of the kit adopt working solutions, thus being convenient to use and low in cost. The method for detecting sunset yellow by using the kit is simple and convenient to operate and has low requirements for pre-processing of samples; and by adopting the method, multitudinous samples can be simultaneously detected quickly. Therefore, the method for detecting sunset yellow by using the ELISA kit can be used for on-site supervision and is suitable for qualitative and quantitative screening of multitudinous samples.

Description

technical field [0001] The invention relates to a method for detecting sunset yellow and an ELISA kit. Background technique [0002] Sunset yellow pigment is one of the six food synthetic pigments approved for use in my country at present. It is mainly used for coloring food and medicine, and can also be used to manufacture aluminum salt lake pigments. China's "Hygienic Standards for the Use of Food Additives" (GB2760-2007) stipulates that sunset yellow can be used in fruit juice drinks, carbonated drinks, concentrated fruit juices, candies, cakes, canned watermelon sauce, puffed shrimp chips, vegetable protein drinks and lactic acid bacteria drinks. The maximum usage amount is 0.10g / kg; for frozen drinks, the maximum usage amount is 0.09g / kg; for candy coating and red and green silk, the maximum usage amount is 0.20g / kg. Its molecular structure is an azo compound, and the products metabolized in the body are aromatic compounds that are potentially harmful to the human body...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543G01N33/535
Inventor 张波王飞郗日沫王鹏易建张霞薛秋艳徐德顺
Owner JIANGSU BERKGEN BIOPHARM
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