Method for measuring activity of superoxide dismutase

A technology for dismutase activity and superoxide, which is applied in the field of determination of superoxide dismutase activity, can solve the problems of difficult detection, low accuracy, and poor reproducibility, and achieve good reproducibility, high accuracy, and easy operation Effect

Active Publication Date: 2014-03-26
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the technical problems of detection difficulty, low accuracy and poor reproducibility in the method for measuring superoxide dismutase activity in the prior art, the present invention provides a method involving ultraviolet-visible spectrophotometry to measure superoxide dismutase active method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A kind of method for measuring superoxide dismutase activity provided by the invention, the method comprises the following steps:

[0037] Step a, preparation of superoxide anion generating system buffer solution

[0038] The preparation concentration is the ammonium bicarbonate buffer solution of 50 mmol / liter, adjusts the pH value to be 9.3;

[0039] Step b, the preparation of standard product

[0040] With the ammonium bicarbonate buffer solution in step a, standard substance is made into the standard solution that concentration is 50 micromole / liter, and described standard substance is 2-(4-iodobenzene)-3-(4-nitrobenzene) -5-(2'4-disulfobenzene)-2H-tetrazolium sodium salt, which can react with superoxide anion to produce water-soluble formazan dye;

[0041] Preparation of step c, reference substance, sample and blank sample

[0042] Preparation of the reference substance: the total reaction volume is 160 microliters, first add 40 microliters of 2 mmol / L EDTA diso...

Embodiment 2

[0061] A kind of method for measuring superoxide dismutase activity provided by the invention, it comprises the following steps:

[0062] Step a, preparation of superoxide anion generating system buffer solution

[0063] The preparation concentration is the ammonium bicarbonate buffer solution of 30 mmol / L, and the pH value is adjusted to 9.5;

[0064] Step b, the preparation of standard product

[0065] With the ammonium bicarbonate buffer solution in step a, standard substance is made into the standard solution that concentration is 50 micromole / liter, and described standard substance is 2-(4-iodobenzene)-3-(4-nitrobenzene) -5-(2'4-disulfobenzene)-2H-tetrazolium sodium salt, which can react with superoxide anion to produce water-soluble formazan dye;

[0066] Preparation of step c, reference substance, sample and blank sample

[0067] Preparation of the reference substance: the total reaction volume is 160 microliters, first add 40 microliters of 4 mmol / L EDTA disodium sa...

Embodiment 3

[0086] A kind of method for measuring superoxide dismutase activity provided by the invention, it comprises the following steps:

[0087] Step a, preparation of superoxide anion generating system buffer solution

[0088] The preparation concentration is the ammonium bicarbonate buffer solution of 45 mmol / L, and the pH value is adjusted to 9.0;

[0089] Step b, the preparation of standard product

[0090] With the ammonium bicarbonate buffer solution in step a, standard substance is made into the standard solution that concentration is 50 micromole / liter, and described standard substance is 2-(4-iodobenzene)-3-(4-nitrobenzene) -5-(2'4-disulfobenzene)-2H-tetrazolium sodium salt, which can react with superoxide anion to produce water-soluble formazan dye;

[0091] Preparation of step c, reference substance, sample and blank sample

[0092] Preparation of the reference substance: the total reaction volume is 160 microliters, first add 40 microliters of 0.4 mmol / L EDTA disodium ...

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Abstract

The invention provides a method for measuring activity of superoxide dismutase. The technical problems of detection difficulty, low accuracy and low reproducibility existing in a method for measuring the activity of superoxide dismutase in the prior art are solved. The method comprises the following steps: selecting a pyrogallic acid alkaline auto-oxidation system, evaluating the removal effect of the superoxide dismutase on superoxide anion by combining an ultraviolet-visible light spectrophotometric method, detecting change of absorbance of a product water-soluble formazan of a trapping agent of sodium 2-(4-iodobenzene)-3-(4-nitrobenzene)-5-(2',4-bisulfophenyl)-2H-tetrazole after acting with the superoxide anion, calculating the removal rate of the superoxide dismutase on the superoxide anion, and further obtaining the activity of superoxide dismutase. The method has the advantages of high reproducibility, high accuracy and simplicity and convenience in operation.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, in particular to a method for measuring the activity of superoxide dismutase. Background technique [0002] Active oxygen includes superoxide anion, hydroxyl radical, hydrogen peroxide, singlet oxygen, etc., and these particles are very small. Due to the presence of unpaired free electrons, reactive oxygen species are chemically very active. Active oxygen is often a by-product of biological and biochemical processes. When it is excessive, the active oxygen in the body will cause damage to cells and gene structures, causing various diseases, such as cytotoxicity, aging, and cancer. Superoxide anion is the source of several other active oxygen species. Superoxide dismutase can remove excess superoxide anion in the body. It can catalyze the disproportionation reaction of superoxide anion to produce oxygen and hydrogen peroxide. It has anti-inflammatory and anti-viral properties. , anti-aging ef...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33
Inventor 刘志强徐晨刘舒邢俊鹏宋凤瑞郑重刘淑莹
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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