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Efficient stable TMB (Tetramethylbenzidine) color-developing liquor and preparation method thereof

A color-developing solution and a stable technology, applied in the field of high-efficiency and stable TMB color-developing solution and its preparation, can solve the problems of short storage time, cumbersome operation, poor stability, etc., to promote dissolution and stability, simplify the operation process, and reduce errors. Effect

Active Publication Date: 2014-04-02
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Common TMB chromogenic reagents are mostly composed of several components, and often need to be prepared and used immediately, which is inconvenient to use and easily leads to unstable test results
Common TMB chromogenic reagents in the general market are usually divided into two stock solutions, A and B, which need to be mixed in proportion to use. The operation is cumbersome and easy to mismatch, which often leads to experimental failure.
In addition, there are rare single-phase TMB chromogenic solutions on the market, but they often have disadvantages such as poor stability, short storage time, high color background, and low sensitivity.

Method used

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  • Efficient stable TMB (Tetramethylbenzidine) color-developing liquor and preparation method thereof
  • Efficient stable TMB (Tetramethylbenzidine) color-developing liquor and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation method of a kind of efficient and stable TMB chromogenic solution of the present invention comprises the steps:

[0022] a. Prepare liquid A: use 10mL dimethyl sulfoxide as solvent, slowly add TMB, stir evenly to fully dissolve.

[0023] b. Preparation of liquid B: Take 1L of distilled water, add 16.406g of sodium acetate to fully dissolve, so that the final molar concentration is 0.2mol / L; then add L(+)-tartaric acid (L(+)-2,3-dihydroxybutyrate Diacid) 4.1g is fully dissolved, so that the final molar concentration is 0.027 mol / L, and the final pH value of the solution is about 5.0.

[0024] c. Add appropriate amount of liquid A, Triton X-405 (Triton X-405), H 2 o 2 and carbamide peroxide, so that the final molar concentration of TMB is 1.2 mmol / L, H 2 o 2 The final molar concentration is 3.1mmol / L, the final mass fraction of Triton X-405 (Triton X-405) is 0.3%, the final mass fraction of carbamide peroxide is 0.5%, and the molar concentration of sod...

Embodiment 2

[0026] The TMB chromogenic solution of Example 1 was compared with the commercialized TMB chromogenic solution of a certain company approved by the State Food and Drug Administration (referred to as comparative chromogenic solution 1).

[0027] The contrast chromogenic solution formula, wherein the molar concentration or mass fraction of each component is:

[0028] The molar concentration of TMB is 2.0mmol / L,

[0029] h 2 o 2 The molar concentration is 4.0mmol / L,

[0030] The mass fraction of carbamide peroxide is 0.1%,

[0031] The molar concentration of disodium hydrogen phosphate is 0.1mol / L,

[0032] The molar concentration of citric acid is 0.013 mol / L.

[0033] Taking the hepatitis C virus core antigen kit (double antibody sandwich ELISA) as an example, the reaction was terminated after 15 minutes of color development, and the OD value (λ=450nm) was measured at 0, 15, 30, 45, and 60 minutes after the reaction was terminated. . After terminating the reaction with a...

Embodiment 3

[0036] The TMB chromogenic solution prepared by the method in Example 1 was compared with the commercial TMB chromogenic solution of a company approved by the State Food and Drug Administration (abbreviated as: comparative chromogenic solution 1).

[0037] Taking the hepatitis C virus core antigen kit (double-antibody sandwich ELISA) as an example, the TMB chromogenic solution prepared by the method in Example 1 and the commercialized TMB chromogenic solution of a company approved by the State Food and Drug Administration were respectively (abbreviated as: contrast chromogenic solution 1) as the chromogenic reagent, and test the hepatitis C core antigen (HCV-cAg) standard (concentration: 80pg / mL). Test every 2 months and record the OD value.

[0038] The results of the comparison are shown in the accompanying drawings of the manual figure 2 . Depend on figure 2 It can be seen that the TMB chromogenic solution prepared by the method of Example 1 can be stored stably for a ...

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Abstract

As a TMB (Tetramethylbenzidine) color-developing liquor on the market has defects of being poor in stability, short in storage time, high in color-developing background, low in sensitivity, and the like, the invention provides an efficient and stable TMB color-developing liquor. The efficient and stable TMB color-developing liquor is characterized by comprising the following components according to molar concentration or by weight percent: TMB with molar with the concentration of 1.2mmol / L, H2O2 with the molar concentration of 3.1mmol / L, 0.3% of trito X-405, 0.5% of urea peroxide, sodium acetate with the molar concentration of 0.2mol / L, L<+>-tartaric acid with the molar concentration of 0.027mol / L. The efficient stable TMB color-developing liquor is a TMB single-phase color-developing liquor, and not required to be mixed before use, so that errors in the artificial operation process can be reduced; a nonionic surfactant is introduced, so that the single-phase TMB color-developing liquor can be stably stored for a long time.

Description

technical field [0001] The invention relates to clinical in vitro detection technology, in particular to an efficient and stable TMB chromogenic solution and a preparation method thereof. Background technique [0002] Since Engvall and Perlman (1971) first reported the establishment of enzyme-linked immunosorbent assays (Enzyme-Linked Immunosorbent Assays, ELISA), due to the advantages of fast, sensitive, simple and easy standardization, ELISA has been rapidly developed and widely used. At present, enzyme immunoassay (EIA) technology has been widely used in the quantitative or qualitative detection and analysis of antigens, haptens or antibodies. Although there are many enzyme-linked immunosorbent assay (ELISA) detection methods, each method is inseparable from enzyme conjugates and chromogenic reagents. The most widely used enzyme in ELISA reagents is horseradish peroxidase (HRP), and there are many kinds of chromogenic reagents in this system, the most commonly used in th...

Claims

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Application Information

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IPC IPC(8): G01N33/531
CPCG01N33/54393G01N33/56983G01N33/5767G01N2333/186
Inventor 谭柏清李志明甘宜梧
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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