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Method for evaluating antioxidant activity of fruit and vegetable food and functional health product

A technology of antioxidant activity and evaluation method, which is applied in the field of evaluation of antioxidant activity and can solve problems such as large differences

Active Publication Date: 2016-05-11
GUANGDONG TIMES FOOD & LIFE HEALTH RES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problem that the current in vitro antioxidant activity analysis and evaluation method directly uses a solvent to extract the antioxidant substances of the sample, which is quite different from the real system of human body digestion and absorption, the present invention establishes a new antioxidant substance based on the cell level. The activity evaluation method, through the extraction of the antioxidant active substances of the tested samples, adopts a multi-enzyme combined enzymatic hydrolysis method that simulates the digestion process of the human body, and then combines the antioxidant activity analysis method at the cellular level, thereby establishing a more biologically relevant , an antioxidant activity evaluation method that is closer to the animal test level

Method used

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  • Method for evaluating antioxidant activity of fruit and vegetable food and functional health product
  • Method for evaluating antioxidant activity of fruit and vegetable food and functional health product
  • Method for evaluating antioxidant activity of fruit and vegetable food and functional health product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Take 10g of Tibetan green 25 highland barley, grind it into powder with a small household powder machine, take 5g of powder, add 100mL of distilled water to make a homogenate, first use 0.1M HCl to adjust the pH to 2.0, then add 20mg of pepsin, at 37°C Under enzymatic hydrolysis for 1.5h. Centrifuge (3000g, 4°C) for 5min, pour off the supernatant A, add 100mL distilled water to the residue, and wash with 0.1M NaHCO 3 Adjust the pH to 7.5, add 15 mg of trypsin, stir at 37 ° C for 5 h, then centrifuge (3000 g, 4 ° C) for 5 min, take the supernatant B; add 100 mL of distilled water to the residue, adjust the pH to 8 with 0.1 M NaOH, and add 2 mg Pronase E and hydrolyze at 37°C for 1 hour, centrifuge, take the supernatant C, add 100mL of distilled water to the residue, adjust the pH to 4, add 50μL Novozymes complex plant hydrolase (ViscozymeL) and stir at 37°C for enzymolysis After 15 hours, centrifuge and take the supernatant D. Collect the supernatants A, B, C and D, co...

Embodiment 2

[0072] Take 10g of Yunnan Changhei highland barley, grind it into powder with a small household powder machine, take 5g of powder, add 120mL of distilled water to make a homogenate, first use 0.1M HCl to adjust the pH to 1.5, then add 25mg of pepsin, at 37 Enzymatic hydrolysis at ℃ for 2h. Centrifuge (3000g, 4°C) for 5min, pour off the supernatant A, add 120mL distilled water to the residue, and use 0.1M NaHCO 3Adjust the pH to 7.0, add 20 mg of trypsin, stir at 37 ° C for 6 h, then centrifuge (3000 g, 4 ° C) for 5 min, take the supernatant B; add 100 mL of distilled water to the residue, adjust the pH to 8.6 with 0.1 M NaOH, and Add 3 mg pronase E and hydrolyze at 37°C for 1.5 h, centrifuge, take the supernatant C, add 120 mL of distilled water to the residue and adjust the pH to 4.5, and add 40 μL of Novozymes compound plant hydrolase (ViscozymeL) and dissolve at 37 Stir the enzymolysis for 12 hours at ℃, centrifuge, and take the supernatant D. Collect the supernatants A, ...

Embodiment 3

[0076] Take 25g of pears, add 125mL of distilled water, use a mixer to beat for 2min, and use a high-speed homogenizer to further beat the material into a homogenate, use 0.1M HCl to adjust the pH to 1.0, then add 15mg of pepsin, at 37°C Enzyme hydrolysis for 1h. Centrifuge (3000g, 4°C) for 5min, pour off the supernatant A, add 100mL distilled water to the residue, and wash with 0.1M NaHCO 3 Adjust the pH to 8.0, add 10 mg of trypsin, stir at 37 ° C for 4.5 h, then centrifuge (3000 g, 4 ° C) for 5 min, take the supernatant B; add 100 mL of distilled water to the residue, and adjust the pH to 8.0 with 0.1 M NaOH, And add 4mg pronase E and enzymolyze at 37°C for 2h, centrifuge, take the supernatant C, add 120mL distilled water to the residue and adjust the pH to 3.5, add 70μL Novozymes compound plant hydrolase (ViscozymeL) and dissolve at 37 Stir the enzymolysis at ℃ for 18 hours, centrifuge, and take the supernatant D. Collect A, B, C and D, concentrate to 50mL, add 50mL ethy...

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Abstract

The invention discloses a method for evaluating the antioxidant activity of fruit and vegetable food and functional health product. The method comprises the following steps: preprocessing a sample to be detected: making a raw material to be detected into water homogenate or directly taking the oral liquid type health food as the sample; co-processing and extracting the sample by multiple enzymes in sequence: processing the sample by pepsin, processing the sample by trypsin, processing the sample by pronase E, and processing the sample by plant composite hydrolase Viscozyme L; removing the reducing sugar in the extract through a cold ethyl acetate extraction technology; and finally analyzing the antioxidant substance content and activity by a conventional method. During the extraction process of substances with antioxidant activity, multiple enzymes are used to carry out enzymatic hydrolysis so as to simulate the human digestion process, at the same time, a cell level antioxidant activity analysis method is used, and thus an antioxidant activity evaluation method, which has stronger biological relevance and is more close to animal experiment level, is established.

Description

technical field [0001] The invention relates to an evaluation method for antioxidant activity, in particular to an evaluation method for antioxidant activity of fruit and vegetable foods and functional health products. The method can be applied to the fields of biomedicine, food, health products and other industries. Background technique [0002] Modern medical research shows that many chronic diseases, including cardiovascular diseases, cancer, diabetes, etc., are closely related to free radicals. Under normal circumstances, the production and elimination of free radicals in the human body are in balance. With age, the activity of antioxidant enzymes decreases, which reduces the ability to scavenge free radicals in the body. Excessive free radicals in the body can damage biological macromolecules (such as proteins, lipids, and DNA), leading to disease and aging. At the same time, external factors such as polluted air, food, unhealthy lifestyle, and emotional factors can al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31G01N1/28
CPCG01N1/28G01N21/3103G01N21/78
Inventor 郑必胜何陵玲朱勇扶雄刘瑞海
Owner GUANGDONG TIMES FOOD & LIFE HEALTH RES CO LTD
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