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Method for rapidly detecting diamine biogenic amine

A diamine, biological amine technology, applied in biochemical equipment and methods, microbial determination/inspection, biological material analysis, etc., can solve the problems of complex steps, expensive instruments, a large amount of organic solvents, etc., to achieve low cost, professional The effect of low requirements and simple operation steps

Pending Publication Date: 2020-08-07
UNIV OF SHANGHAI FOR SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Putrescine mostly exists in spoiled food, and the increase of putrescine will not only have a negative impact on the flavor, but also pose a potential threat to food safety and human health
Eating foods high in putrescine can lead to serious toxicological consequences
[0004] In recent years, many analytical methods have been developed to detect such biogenic amines. The most widely used methods are high performance liquid chromatography, capillary gas chromatography, capillary zone electrophoresis, ion exchange chromatography, etc. The advantages of these analytical methods are The separation effect is good and the detection limit is low, but the disadvantages are also obvious, such as expensive instruments, professional operation, complicated steps, and the need for a large amount of organic solvents, etc.

Method used

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  • Method for rapidly detecting diamine biogenic amine
  • Method for rapidly detecting diamine biogenic amine
  • Method for rapidly detecting diamine biogenic amine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The detection method of diamine biogenic amine comprises the steps:

[0031] 1) Preparation of diamine oxidase solution: use PBS buffer solution with a concentration of 0.05-1mol / L and a pH of 7.2 to prepare an enzyme solution with a concentration of 0.1-10mg / mL;

[0032] 2) Make the chromogenic solution: 1mL of the chromogenic solution (prepared with double distilled water) contains 0.5mg of tetramethylbenzidine;

[0033] 3) Making a reaction mixture solution: add 100 μL of diamine oxidase solution to 100 μL of a certain concentration of diamine biogenic amine solution;

[0034] 4) React the mixed solution at 50°C for 25 minutes to start the reaction, add 50 μL of 0.1-10 mg / mL HRP solution, 200 μL of 0.5 mg / mL TMB color development solution for color development, and then add 50 μL of 2mol / L dilute Hydrochloric acid terminated the reaction.

[0035] 5) Measure the absorbance value at 450 nm with a spectrophotometer.

[0036] A series of histamine solutions and putre...

Embodiment 2

[0041] A method for rapidly detecting diamine biogenic amines, the method comprising the following steps:

[0042] 1) After mixing the diamine biogenic amine solution and the diamine oxidase solution, react at 45° C. for 30 minutes. Among them, the preparation process of the diamine oxidase solution is: add diamine oxidase to the PBS buffer solution with a concentration of 0.05mol / L and a pH of 7.5 to prepare a solution with a concentration of 0.1mg / mL; The volume ratio of the amine solution to the diamine oxidase solution is 100:120.

[0043] 2) Adding horseradish peroxidase solution and TMB color developing solution, adding dilute hydrochloric acid to terminate the reaction after developing the color, and obtaining a sample. Wherein, the concentration of horseradish peroxidase solution is 0.1mg / mL, the concentration of TMB chromogenic solution is 0.6mg / mL, and the concentration of dilute hydrochloric acid is 1.8mol / L; Horseradish peroxidase solution, TMB chromogenic solutio...

Embodiment 3

[0046] A method for rapidly detecting diamine biogenic amines, the method comprising the following steps:

[0047] 1) After mixing the diamine biogenic amine solution and the diamine oxidase solution, react at 55° C. for 20 minutes. Among them, the preparation process of the diamine oxidase solution is as follows: add diamine oxidase to PBS buffer solution with a concentration of 1mol / L and a pH of 7 to prepare a solution with a concentration of 10mg / mL; diamine biogenic amine solution The volume ratio with diamine oxidase solution is 100:80.

[0048]2) Adding horseradish peroxidase solution and TMB color developing solution, adding dilute hydrochloric acid to terminate the reaction after developing the color, and obtaining a sample. Wherein, the concentration of horseradish peroxidase solution is 10mg / mL, the concentration of TMB chromogenic solution is 0.4mg / mL, and the concentration of dilute hydrochloric acid is 2.2mol / L; Horseradish peroxidase solution, TMB chromogenic s...

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Abstract

The invention relates to a method for quickly detecting the diamine biogenic amine. The method comprises the following steps of: 1) mixing a diamine biogenic amine solution with a diamine oxidase solution, and carrying out a reaction at 45-55 DEG C for 20-30 minutes; 2) adding a horse radish peroxidase solution and a TMB color developing solution, and adding diluted hydrochloric acid to terminatethe reaction after color development to obtain a sample; and 3) placing the sample in a spectrophotometer to detect an absorbance value so as to obtain the concentration of the diamine biogenic aminein the diamine biogenic amine solution. Compared with the prior art, the method has the advantages of simple operation process and low professional requirements; experimental results can be directly judged by naked eyes, which enables simplicity and clearness; and results can be obtained quickly within about 25 min in a whole process, and so that the method is suitable for on-site detection.

Description

technical field [0001] The invention belongs to the technical field of biogenic amine detection and relates to a method for rapidly detecting diamine biogenic amines. Background technique [0002] Biogenic amines are a class of non-volatile and biologically active low-molecular nitrogen-containing organic bases. According to the number of amino groups, they can be divided into monoamines and diamines. Among them, diamine biogenic amines mainly include histamine and putrescine. Biogenic amines are mainly formed by free amino acids under the catalysis of decarboxylase in bacteria, so there are often a certain amount of biogenic amines in foods rich in amino acids, such as dairy products, fermented wine, pickled aquatic products and meat products, etc. . [0003] Low concentrations of biogenic amines are normal active ingredients in organisms and have important physiological functions in biological cells, but ingestion of high concentrations of biogenic amines has toxic effect...

Claims

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Application Information

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IPC IPC(8): G01N21/78C12Q1/28C12Q1/26
CPCC12Q1/26C12Q1/28C12Q2326/12G01N21/78G01N2333/90638G01N2333/908
Inventor 徐斐袁敏曹慧叶泰于劲松
Owner UNIV OF SHANGHAI FOR SCI & TECH
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