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Method for evaluating antioxidant activity of antioxidant substance

A technology of anti-oxidation activity and anti-oxidation substances, applied in the biological field, can solve the problems of increased difficulty in operation, sensitivity to temperature and pH value of fluorescent substances, expensive instruments, etc., and achieve the effect of overcoming the long cycle

Inactive Publication Date: 2017-12-08
SOUTHWEST UNIVERSITY
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Problems solved by technology

This method is a novel anti-oxidation evaluation method. Its main disadvantages are that the required instruments are relatively expensive, and the fluorescent substance is very sensitive to temperature and pH value, which makes the operation of the experiment more difficult.
[0010] At present, there is no report of Saccharomyces cerevisiae as a model to evaluate antioxidant activity. Therefore, the present invention uses yeast as a model organism to establish a yeast oxidative stress model to study the effect of antioxidant substances on Saccharomyces cerevisiae under oxidative stress conditions. Antioxidant substances Antioxidant activity evaluation provides a new method

Method used

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  • Method for evaluating antioxidant activity of antioxidant substance
  • Method for evaluating antioxidant activity of antioxidant substance
  • Method for evaluating antioxidant activity of antioxidant substance

Examples

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Embodiment 1

[0059] The oxidative stress treatment of embodiment 1 Saccharomyces cerevisiae

[0060] Pick a single colony of yeast and culture it in liquid to OD 600 = 1.0 or so, as bacterial strains, inoculated in a 100 mL Erlenmeyer flask containing 20 mL of YPD liquid medium at a ratio of 0.1%, 0.5%, and 1.0%, respectively. Join in groups to prepare H 2 o 2 , so that the final concentration is 0mM (control), 0.2mM, 1.0mM, 2.0mM.

[0061] The ATCC9763 and S288c bacterial liquids were used as the parent species to do the above treatment, and different concentrations of H 2 o 2 Inhibitory effects of treatments on different strains of yeast.

[0062] Group A model: H immediately after S. cerevisiae inoculation 2 o 2 oxidative stress

[0063] by OD 600 =1.0 Yeast (ATCC9763 and S288c) bacterial solution is the mother species, inoculate according to different proportions (0.1%, 0.5%, 1.0%), and add H at the same time 2 o 2 treatment, observing different concentrations of H 2 o 2In...

Embodiment 2

[0072] Example 2 Using the evaluation method of antioxidant activity of antioxidant substances to evaluate the antioxidant effect of quercetin

[0073] 1. The effect of auxiliary agent DMSO on the growth of yeast ATCC9763

[0074] The yeast with 1.0% inoculum ratio treated with 0.1% DMSO was used as the treatment group to study the effect of the auxiliary agent DMSO on the growth of yeast ATCC9763 when the volume fraction was 0.1%. Depend on Figure 5 It can be seen that the growth curves of the treatment group (0.1% DMSO) and the control group almost completely overlap. The experimental results showed that 0.1% DMSO neither inhibited nor promoted the growth of yeast ATCC9763. Therefore, it need not be considered as an experimental factor in the growth curve determination.

[0075] 2. Screening of Quercetin Concentration Gradients

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to an evaluation method for the antioxidant activity of antioxidant substances. This evaluation method takes Saccharomyces cerevisiae as a model, inoculates and cultures Saccharomyces cerevisiae, and then adds oxidant oxidative stress culture to obtain the Saccharomyces cerevisiae oxidative stress model, which is the control group; adds the test sample to the inoculated Saccharomyces cerevisiae, and then adds oxidant oxidation stress culture. The stress culture was used to obtain the treatment group; the antioxidant activity of the test samples was evaluated by measuring and comparing the growth of Saccharomyces cerevisiae and the antioxidant index in Saccharomyces cerevisiae in the control group and the treatment group. The evaluation method provided by the invention is close to the actual system of living organisms, and is relatively sensitive. Taking the brewer's yeast as a model, it overcomes the shortcomings of long period, high cost and cumbersome experimental process. The present invention uses bacterial strain ATCC9763 as a model to establish an evaluation of antioxidant activity, and provides a research method for exploring the antioxidant activity of antioxidant substances in vivo.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an evaluation method for the antioxidant activity of antioxidant substances. Background technique [0002] Currently, there are many methods used to study antioxidant capacity. Common in vitro antioxidant assay methods mainly include the following: DPPH (1,1-diphenyl-2-picrylhydrazyl radical) method, ABTS (2,2'-azinobis (3-ethbenzothiazoline-6-sulfonate) method, FRAP ( Ferric reducing antioxidant power) method, ORAC (oxygen radical absorbance capacity) oxygen free radical absorption capacity, cell antioxidant method, P-carotene bleatene bleching method (P-carotene linoleic acid system) and low-density lipoprotein (LDL) (low-density lipoprotein (LDL) Density lipoprotein oxidation reaction), etc. Among them, DPPH, ABTS, FRAP and ORAC are methods for measuring the ability of antioxidants to scavenge free radicals, and have been widely used in the evaluation of the antioxida...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12R1/865
CPCC12Q1/025G01N2333/395
Inventor 潘宇周志钦胡定辉徐晓丹柯尊丽聂超
Owner SOUTHWEST UNIVERSITY
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