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tmb two-component chromogenic solution and its kit

A chromogenic solution and two-component technology, applied in the field of enzyme-linked immunoassay, can solve the problems of low safety, reduced reaction sensitivity, easy mixing and unevenness, etc.

Active Publication Date: 2020-08-25
CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Based on this, it is necessary to provide a TMB two-component chromogenic solution and a kit thereof for the traditional two-component TMB chromogenic solution with low safety, easy mixing and unevenness, and reduced reaction sensitivity.

Method used

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  • tmb two-component chromogenic solution and its kit
  • tmb two-component chromogenic solution and its kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Weigh 0.20 g of disodium edetate, 1.90 g of citric acid, 50 mL of glycerin, and 1.0 g of vitamin C into a volumetric flask with an appropriate amount of double-distilled water 1 L in advance. After fully dissolving, add tetramethylbenzidine hydrochloride 0.20g, dilute to 1L, and mix well to obtain TMB stock solution.

[0038] Weigh 0.60 g of carbamide peroxide, 9.40 g of citric acid, and 18.60 g of disodium hydrogen phosphate dodecahydrate, place them in a volumetric flask with an appropriate amount of double-distilled water 1 L in advance, dissolve them fully and set the volume to 1 L, and mix well to obtain TMB substrate buffer.

[0039] Replace the TMB chromogenic solution in the aflatoxin M1 detection kit purchased from Shenzhen Fender Biotechnology Co., Ltd. with the prepared TMB stock solution and TMB substrate buffer solution, and measure the sensitivity based on the above reagents according to the kit usage method , minimum detection limit, accuracy, precision ...

Embodiment 2

[0057] Weigh 0.18g of disodium edetate, 1.52g of citric acid, 40mL of glycerin, and 0.18g of vitamin C into a volumetric flask with an appropriate amount of double-distilled water 1L in advance. After fully dissolving, add tetramethylbenzidine hydrochloride 0.18g, dilute to 1L, and mix well to obtain TMB stock solution.

[0058] Weigh 0.48g of carbamide peroxide, 7.52g of citric acid, and 14.88g of disodium hydrogen phosphate dodecahydrate, and place them in a volumetric flask with an appropriate amount of double-distilled water 1L in advance. After fully dissolving, set the volume to 1L, and mix well to obtain TMB substrate buffer.

[0059] Sensitivity, minimum detection limit, accuracy, precision and stability experiments were performed with the same method as in Example 1, and the experimental results are shown in Table 2.

Embodiment 3

[0061] Weigh 0.22g of disodium edetate, 2.28g of citric acid, 60mL of glycerin, and 0.22g of vitamin C into a volumetric flask with an appropriate amount of double-distilled water 1L in advance. After fully dissolving, add tetramethylbenzidine hydrochloride 0.22g, dilute to 1L, mix well, and prepare TMB stock solution.

[0062] Weigh 0.72g of carbamide peroxide, 11.28g of citric acid, and 22.32g of disodium hydrogen phosphate dodecahydrate into a volumetric flask with an appropriate amount of double-distilled water 1L in advance, dissolve fully and set the volume to 1L, and mix well to obtain TMB substrate buffer.

[0063] Sensitivity, minimum detection limit, accuracy, precision and stability experiments were performed with the same method as in Example 1, and the experimental results are shown in Table 2.

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Abstract

The invention provides a double-component TMB (3,3',5,5'-tetramethyl benzidine) color developing solution. The color developing solution comprises a TMB stock solution and a TMB substrate buffer solution, and water serves as a solvent of the TMB stock solution; every 1,000 mL of the TMB stock solution contains 0.18-0.22 g of disodium EDTA, 1.52-2.28 g of citric acid, 40-60 mL of glycerin, 0.18-0.22 g of tetramethyl benzidine and 0.80-1.20 g of vitamin C. The TMB stock solution takes hydrochloride of TMB as a color developing agent, and the prepared TMB stock solution can exist in the form of aqueous solution. The vitamin C can improve the stability of TMB, the TMB stock solution and the TMB substrate buffer solution can be easily and uniformly mixed in a mixing chamber, and the differencebetween batches is reduced; the vitamin C can improve the stability of TMB, the content of effective color developing components is guaranteed after mixing, and the sensitivity in the color developingprocess is improved.

Description

technical field [0001] The invention relates to an enzyme-linked immunoassay, in particular to a TMB two-component chromogenic solution and a kit with the same. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) has the advantages of rapidity, sensitivity, simplicity, and easy standardization. Although there are many kinds of ELISA, all methods are inseparable from enzyme conjugates and chromogenic reagents. The most widely used enzyme in ELISA reagents is horseradish peroxidase (HRP), and the most commonly used corresponding chromogenic reagent is 3,3',5,5'-tetramethylbenzidine (3,3', 5,5'-Tetramethyl Benzidine, TMB), a blue substance will be produced after color development, and the stop solution is added to terminate the color reaction, and the solution changes from blue to yellow, and the color intensity is proportional to the antibody content. During the detection process, use a microplate reader to read at a wavelength of 450nm / 630nm, record the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
Inventor 骆鹏杰王华丽陈霞赵云峰李敬光蒋双勤李湖中
Owner CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT
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