CN113298381A - Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia - Google Patents

Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia Download PDF

Info

Publication number
CN113298381A
CN113298381A CN202110575404.3A CN202110575404A CN113298381A CN 113298381 A CN113298381 A CN 113298381A CN 202110575404 A CN202110575404 A CN 202110575404A CN 113298381 A CN113298381 A CN 113298381A
Authority
CN
China
Prior art keywords
antioxidant
ganoderma lucidum
absorbance
evaluation index
water extract
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202110575404.3A
Other languages
Chinese (zh)
Inventor
韩伟
郑丹婷
陈炼茹
冯杰
冯娜
卜原玲
姚泽远
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
East China University of Science and Technology
Original Assignee
East China University of Science and Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by East China University of Science and Technology filed Critical East China University of Science and Technology
Priority to CN202110575404.3A priority Critical patent/CN113298381A/en
Publication of CN113298381A publication Critical patent/CN113298381A/en
Pending legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06Q—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES; SYSTEMS OR METHODS SPECIALLY ADAPTED FOR ADMINISTRATIVE, COMMERCIAL, FINANCIAL, MANAGERIAL OR SUPERVISORY PURPOSES, NOT OTHERWISE PROVIDED FOR
    • G06Q10/00—Administration; Management
    • G06Q10/06—Resources, workflows, human or project management; Enterprise or organisation planning; Enterprise or organisation modelling
    • G06Q10/063—Operations research, analysis or management
    • G06Q10/0639—Performance analysis of employees; Performance analysis of enterprise or organisation operations
    • G06Q10/06393—Score-carding, benchmarking or key performance indicator [KPI] analysis
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06F—ELECTRIC DIGITAL DATA PROCESSING
    • G06F17/00—Digital computing or data processing equipment or methods, specially adapted for specific functions
    • G06F17/10—Complex mathematical operations
    • G06F17/18—Complex mathematical operations for evaluating statistical data, e.g. average values, frequency distributions, probability functions, regression analysis

Landscapes

  • Engineering & Computer Science (AREA)
  • Business, Economics & Management (AREA)
  • Human Resources & Organizations (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Data Mining & Analysis (AREA)
  • Theoretical Computer Science (AREA)
  • Strategic Management (AREA)
  • Pure & Applied Mathematics (AREA)
  • Mathematical Physics (AREA)
  • Operations Research (AREA)
  • Mathematical Optimization (AREA)
  • Mathematical Analysis (AREA)
  • Entrepreneurship & Innovation (AREA)
  • Development Economics (AREA)
  • Economics (AREA)
  • Educational Administration (AREA)
  • Computational Mathematics (AREA)
  • Game Theory and Decision Science (AREA)
  • Quality & Reliability (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Business, Economics & Management (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Tourism & Hospitality (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Probability & Statistics with Applications (AREA)
  • Evolutionary Biology (AREA)
  • Algebra (AREA)
  • Marketing (AREA)
  • Databases & Information Systems (AREA)
  • Software Systems (AREA)
  • General Engineering & Computer Science (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Cosmetics (AREA)

Abstract

The invention relates to a method for comprehensively evaluating the antioxidant activity of ganoderma lucidum mycelia, which comprises the following steps: (1) preparing water extract from ganoderma lucidum mycelia; (2) measuring various antioxidant evaluation index values of the water extract respectively; (3) weighting the antioxidant evaluation index value in the step (2) by an entropy weight method; (4) and (3) calculating to obtain a comprehensive antioxidant activity Z value of the ganoderma lucidum mycelia according to the antioxidant evaluation index value in the step (2) and the weight obtained in the step (3). According to the invention, by establishing a comprehensive evaluation method of the antioxidant activity of the ganoderma lucidum mycelia, the difference and the change condition of the in-vitro antioxidant activity of different ganoderma lucidum mycelia extracts can be objectively and comprehensively evaluated, and a method for associating and assigning weights to a plurality of indexes can objectively and truly reflect the antioxidant capacity of the ganoderma lucidum mycelia under each evaluation index, so that the method can be widely applied to the evaluation of the antioxidant activity of natural products.

Description

Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia
Technical Field
The invention relates to a comprehensive evaluation method of antioxidant activity of natural products, in particular to a method for comprehensively evaluating the in-vitro antioxidant activity of ganoderma lucidum mycelium aqueous extract.
Background
Ganoderma lucidum (Ganoderma lucidum) is a rare medicinal fungus distributed in yellow river and Yangtze river basin in China, Japan, Europe and parts of North America, and has abundant resource types. Modern researches show that ganoderma lucidum has various pharmacological activities of inducing cancer cell apoptosis, inhibiting inflammatory reaction, resisting oxidation, regulating body immunity, preventing lipid metabolism disorder, protecting nerves and the like.
The medicinal value of the ganoderma lucidum causes the ganoderma lucidum to have great market demand, the ganoderma lucidum in the traditional sense is the fruiting body of the ganoderma lucidum or the ganoderma sinensis of the polyporaceae, the fruiting body is influenced by factors such as regions, seasons, raw materials and the like, the quality control is poor, the growth cycle is long, the cost is high, and the commercial application of the ganoderma lucidum is limited. The ganoderma lucidum mycelia are the basic structure of the ganoderma lucidum nutriment, can be quickly obtained by a liquid fermentation technology, can obtain mycelia containing more target components (such as triterpenes, polysaccharides, proteins, amino acids, alkaloids and the like) by adjusting culture conditions, and are beneficial to large-scale and standardization of industrial production of ganoderma lucidum effective components.
Medical research shows that many important human diseases such as atherosclerosis, rheumatoid arthritis, diabetes, cancer and aging process are related to oxidative damage caused by free radicals, so research and development of antioxidant active substances become hot spots for researchers at home and abroad. Chemically synthesized antioxidants, such as: dibutylhydroxytoluene (BHT), Butylhydroxyanisole (BHA), etc. have been once favored, but animal experiments have found that they have certain toxic and teratogenic effects. The natural antioxidant is safe and nontoxic, and meets the new requirements of people on health and safety.
In vitro antioxidant research on natural products is numerous, a standardized evaluation method is not available at present, and because the antioxidant mechanism is complex, multiple methods are usually selected for evaluation respectively, so that results obtained by different methods need to be integrated by using a multi-index analysis method. The multi-index comprehensive analysis method can be divided into a subjective method and an objective method according to a weight calculation method. The entropy weight method is an objective method for weighting according to the respective discrimination of different evaluation indexes, the evaluation result of the index weight determined by the method has strong mathematical theoretical basis, irrationality of subjective judgment is avoided, weight calculation conforming to objective practice is truly achieved, the calculation mode is simple, operability is strong, and comprehensive evaluation of results obtained by various methods is facilitated.
At present, no research is found at home and abroad for comprehensively evaluating the in-vitro antioxidant activity of the edible fungus extract by using an entropy weight method.
Disclosure of Invention
One of the purposes of the invention is to provide a novel method which is simple in calculation mode, strong in operability and beneficial to comprehensive evaluation of the antioxidant activity of ganoderma lucidum mycelia.
The method provided by the invention has the advantages that the method overcomes the limitation that only a specific ingredient in the lucid ganoderma can be subjected to antioxidant activity evaluation in the prior art and only the same evaluation method can be subjected to antioxidant comparative analysis in the prior art, the water-soluble antioxidant ingredient of the lucid ganoderma is taken as an evaluation object, the comprehensive antioxidant activity is taken as an evaluation index, and the comprehensive antioxidant activity is calculated by an entropy weight method to obtain a simple and intuitive evaluation method, so that the antioxidant activity of the lucid ganoderma is comprehensively and objectively evaluated.
In order to achieve the purpose, the specific technical scheme of the invention is as follows:
a method for comprehensively evaluating the antioxidant activity of ganoderma lucidum mycelia comprises the following steps:
(1) preparing water extract from ganoderma lucidum mycelia;
(2) respectively measuring various antioxidant evaluation index values of the water extract;
(3) weighting the antioxidant evaluation index value in the step (2) by an entropy weight method;
(4) and (3) calculating to obtain a comprehensive antioxidant activity Z value of the ganoderma lucidum mycelia according to the antioxidant evaluation index value in the step (2) and the weight obtained in the step (3).
In the step (2), the antioxidant evaluation index value is DPPH free radical clearance rate, hydroxyl free radical clearance rate and/or ABTS free radical clearance rate.
In the step (1), the preparation conditions of the water extract are as follows: precisely weighing 0.05-2 g of ganoderma lucidum mycelium solid, carrying out ultrasonic water extraction under the conditions of liquid-solid ratio of 20-70 mL/g, ultrasonic temperature of 30-60 ℃, ultrasonic time of 15-75 min and ultrasonic power of 72-180W, centrifuging the solution after extraction, and taking a proper amount of supernatant to prepare 1-2 mg/mL (mg/mL is the mass of ganoderma lucidum mycelium powder before extraction/the mass of solvent after preparation of the solution) of ganoderma lucidum mycelium extract.
In the step (2), the method for measuring the DPPH free radical clearance rate comprises the following steps: placing 2mL of the water extract in a brown volumetric flask, adding 2mL of DPPH free radical working solution with the absorbance of 0.7 +/-0.01, reacting for 30min in a dark place, measuring the absorbance at the position of 517nm, and adjusting to zero by using 95% ethanol;
then, DPPH radical clearance S was calculated according to equation (1)1Repeating the steps for three times, and taking an average value:
Figure BDA0003083381990000031
wherein A isiIs the absorbance, A, of the water extract after reaction with the DPPH free radical working solutionjThe background absorbance of the aqueous extract, measured using 95% ethanol instead of the DPPH free radical working solution, A0The blank absorbance was measured by replacing the aqueous extract with deionized water.
In the step (2), the method for measuring the hydroxyl radical clearance rate of the water extract comprises the following steps: putting 1mL of the water extract into a test tube with a plug, and sequentially adding 1mL of 9mmol/L FeSO4And 8.8mmol/L H2O2Standing the solution at room temperature for 10min, transferring into 1mL 9mmol/L salicylic acid-ethanol solution, mixing, maintaining in 37 deg.C water bath for 30min, and allowing the water extractive solution to react with salicylic acid (H)2O2With FeSO4The hydroxyl free radical generated by the reaction is captured by salicylic acid to generate 2, 3-dihydroxybenzoic acid which is obviously absorbed at 510nm, namely the salicylic acid reaction system) to be fully reactedAfter the reaction, the solution was centrifuged, and the absorbance A of the supernatant at 510nm was measuredi' zeroing with deionized water;
then, the hydroxyl radical clearance S was calculated according to the formula (2)2Repeating the steps for three times, and taking an average value:
Figure BDA0003083381990000032
wherein A isj' is to replace H with deionized water2O2Measuring the background absorbance of the water extract, A0' is the blank absorbance measured after the deionized water is used for replacing the water extract;
the salicylic acid reaction system refers to the reaction system H2O2With FeSO4The hydroxyl radical generated by the reaction is captured by salicylic acid to generate 2, 3-dihydroxybenzoic acid which has obvious absorption at 510 nm.
In the step (2), the method for determining the ABTS free radical clearance of the water extract comprises the following steps: adding 1mL of the water extract into a brown volumetric flask, adding 4mL of ABTS free radical working solution with the absorbance near 0.7, shaking up, reacting for 6min in a dark place, and measuring the absorbance of the solution at 734 nm;
then, ABTS radical clearance S was calculated according to equation (3)3The experiment was repeated three times, and the mean value was taken:
Figure BDA0003083381990000033
wherein A isi"is the absorbance, A, of the water extract after reaction with the ABTS free radical working solutionj"is the background absorbance of the sample solution measured after replacing the ABTS free radical working solution with phosphate buffer solution, A0"is the blank absorbance measured after the deionized water is used to replace the water extract.
The preparation method of the ABTS free radical working solution comprises the following steps: and (3) transferring 5mL of 7mmol/L ABTS solution and 100 mu L of 140mmol/L potassium persulfate solution into a 10mL brown volumetric flask, shaking uniformly, reacting for 12-16h in a dark place, and diluting by about 90 times by using a phosphate buffer solution with the pH value of 7.4 to ensure that the absorbance is about 0.7, thereby obtaining the ABTS free radical working solution.
In the step (3), the method for giving the weight is as follows:
first, the antioxidant ability s of the water extract according to the formula (4)ijCarrying out dispersion standardization calculation to obtain yij；
Figure BDA0003083381990000041
Wherein s isijThe j-th antioxidant evaluation index value of the i-th extracting solution is 1, 2, 3, …, m, j is 1, 2, 3, …, n; y isijIs s isijValues normalized by dispersion; for example, DPPH radical clearance is j ═ 1, hydroxyl radical clearance is j ═ 2, ABTS radical clearance is j ═ 3;
then, the information entropy E is calculated as equation (5)j；
Figure BDA0003083381990000042
Finally, the weight w is calculated according to equation (6)j：
Figure BDA0003083381990000043
Wherein the content of the first and second substances,
Figure BDA0003083381990000044
is the proportion of the result of the ith group of experimental data under the j-th antioxidant evaluation index value, and when p isijWhen 0, define
Figure BDA0003083381990000045
yijIs s in formula (4)ijObtaining a numerical value after dispersion standardization; kappa is the antioxidant evaluation index value; m is the number of experimental data sets; 1nm, lnpijRespectively represent m and pijNatural logarithm of (E)jInformation entropy, w, of the j-th antioxidant assessment index valuejThe weight of the index value of the i-th antioxidant assessment.
In the step (4), the calculation method of the comprehensive antioxidant activity Z value comprises the following steps: z is S1×w1+S2×w2+S3×w3(ii) a Wherein S is1Is DPPH radical clearance, S2Is hydroxy radical scavenging rate, S3For ABTS radical clearance, w1、w2、w3Are respectively S1、S2、S3The corresponding weight.
The centrifugation condition is centrifugation for 3-15min under the conditions of 6000-12000 rpm.
The invention has the beneficial effects that: the invention provides an objective and comprehensive multi-index comprehensive evaluation method for antioxidant activity, namely, an entropy weight method is utilized to solve information entropies of different indexes, and then weight calculation is carried out, the limitation that only a single antioxidant index can be evaluated and multi-index subjective weighting in the prior art is overcome, and the method can effectively, truly, objectively and comprehensively carry out comparative analysis on the antioxidant activity of lucid ganoderma, and further can be popularized to antioxidant activity screening and multi-index process optimization of edible fungi.
Detailed Description
The following examples of the present invention will be described in detail, but the description should not be construed as limiting the scope of the present invention. All equivalent changes, modifications and the like made within the scope of the present invention shall fall within the protection and coverage of the present invention. In addition, it should be noted that the embodiments and features of the embodiments of the present invention may be combined with each other without conflict.
Example 1
A method for comprehensively evaluating the antioxidant activity of ganoderma lucidum mycelia comprises the following operations:
taking 0.05g of each ganoderma lucidum mycelium in the following table 1, respectively adding 2mL of deionized water, shaking up, performing ultrasonic treatment for 50min under the condition of water bath at 40 ℃, centrifuging the obtained water extract for 12min under the condition of 11000r/min, taking 1mL of supernatant into a 25mL volumetric flask, and complementing the volume to the scale with deionized water to obtain 74 parts of each ganoderma lucidum mycelium sample solution.
TABLE 1
Figure BDA0003083381990000051
Figure BDA0003083381990000061
Figure BDA0003083381990000071
Detecting DPPH free radical clearance, hydroxyl free radical clearance and ABTS free radical clearance of each ganoderma lucidum mycelium sample solution to respectively obtain S1、S2、S3The data are listed in table 2. Because the DPPH method, the hydroxyl radical method and the ABTS method relate to different antioxidant mechanisms, as can be seen from the following Table 2, the scavenging conditions of the same ganoderma lucidum mycelium material for different free radicals are different, and the weights calculated by the entropy weight method for different evaluation methods are sequentially as follows: 0.6087(DPPH method), 0.1397 (hydroxyl radical method) and 0.2516(ABTS method).
The entropy weight method calculates the weight as follows:
first, the antioxidant ability s of the water extract according to the formula (1)ijCarrying out dispersion standardization calculation to obtain yij；
Figure BDA0003083381990000072
Wherein s isijThe data (i: 1, 2, 3, …, 70; j: 1, 2, 3) for the j-th evaluation index of the extract liquid numbered i in Table 1 are, for example, DPPH radical scavenging rate (j: 1), hydroxyl radical scavenging rate (j: 2), ABTS radical scavenging rate (j: 3), yijIs s isijValues normalized by dispersion.
Then, the information entropy E is calculated according to equation (2)j；
Figure BDA0003083381990000081
Wherein the content of the first and second substances,
Figure BDA0003083381990000082
is the proportion of the clearance rate of free radicals of ganoderma lucidum mycelia with the serial number i under the j item antioxidant evaluation index value, and when p isijWhen 0, define
Figure BDA0003083381990000083
yijIs s in formula (1)ijObtaining a numerical value after dispersion standardization; 1nm is the natural logarithm of m (logarithm based on constant e), lnpijIs pijThe natural logarithm of (c). m is the number of types of the ganoderma lucidum mycelia in Table 1 (i.e., in this example, m is 70)
Finally, the weight w is calculated according to equation (3)j：
Figure BDA0003083381990000084
κ is the antioxidant evaluation index value (in this example, κ ═ 3); ejInformation entropy, w, of the j-th antioxidant assessment index valuejThe weight of the j-th antioxidant evaluation index value.
Calculated by the formula (3): w is a1＝0.6087、w2＝0.1397、w3＝0.2516。
In summary, the method for calculating the comprehensive antioxidant activity Z value in this example is as follows: z is S1×0.6087+S2×0.1397+S3X 0.2516, and the combined antioxidant activities Z are listed in Table 2.
TABLE 2
Figure BDA0003083381990000085
Figure BDA0003083381990000091
Figure BDA0003083381990000101
Analyzing the data in table 2, it can be seen that G0119, which has the highest hydroxyl radical clearance rate, has weaker DPPH radical clearance rate and ABTS radical clearance rate; g0149, which has the highest ABTS free radical scavenging rate, has poor hydroxyl radical scavenging effect although the DPPH free radical scavenging rate is also high; g0019 with the highest DPPH free radical scavenging rate has higher ABTS free radical scavenging activity and poorer hydroxyl free radical scavenging activity. Therefore, the optimal strain results obtained according to different evaluation methods are different.
According to weight calculation, the weight of the three free radical scavenging effects is DPPH free radical scavenging method > ABTS free radical scavenging method > hydroxyl free radical scavenging method from large to small, the weight of the hydroxyl free radical scavenging method is the minimum, and G0149 is the ganoderma lucidum mycelium with the best antioxidant activity according to comparison of comprehensive antioxidant activity Z. The experimental result is more comprehensive and objective.
Example 2
A multi-index analysis method for optimizing an extraction process of antioxidant active substances of ganoderma lucidum mycelia comprises the following operations:
the four-factor five-level star point design based on the single-factor test-the effect surface factor level table is shown in table 3:
TABLE 3
Figure BDA0003083381990000111
Preparing water extractive solution of Ganoderma mycelia according to the conditions listed in Table 4 below, measuring antioxidant activity of each extractive solution, measuring each solution for three times, and averaging to obtain S1、S2、S3。
As the star point design-effect surface experiment can only be fitted aiming at a single index, the entropy weight method is adopted to carry out weight calculation on the experiment results of different evaluation methods under each experiment condition, and the method sequentially comprises the following steps: 0.3054(DPPH method), 0.4030 (hydroxyl radical), 0.2916(ABTS method).
The entropy weight method calculates the weight as follows:
first, the antioxidant ability s of the water extract according to the formula (1)ijCarrying out dispersion standardization calculation to obtain yij；
Figure BDA0003083381990000112
Wherein s isijThe data (i: 1, 2, 3, …, 30; j: 1, 2, 3) for the j-th evaluation index of the extract liquid numbered i in Table 4 are, for example, DPPH radical scavenging rate (j: 1), hydroxyl radical scavenging rate (j: 2), ABTS radical scavenging rate (j: 3), yijIs s isijValues normalized by dispersion.
Then, the information entropy E is calculated according to equation (2)j；
Figure BDA0003083381990000121
Wherein the content of the first and second substances,
Figure BDA0003083381990000122
is the proportion of the clearance rate of free radicals of ganoderma lucidum mycelia with the serial number i under the j item antioxidant evaluation index value, and when p isijWhen 0, define
Figure BDA0003083381990000123
yijIs s in formula (1)ijObtaining a numerical value after dispersion standardization; 1nm is the natural logarithm of m (logarithm based on constant e), lnpijIs pijThe natural logarithm of (c). m is the number of types of the ganoderma lucidum mycelia in Table 1 (i.e., m is 30 in this example)
Finally, the weight w is calculated according to equation (3)j：
Figure BDA0003083381990000124
κ is the antioxidant evaluation index value (in this example, κ ═ 3); ejInformation entropy, w, of the j-th antioxidant assessment index valuejThe weight of the j-th antioxidant evaluation index value.
Calculated by the formula (3): w is a1＝0.3054、w2＝0.4030、w3＝0.2916。
In summary, the method for calculating the comprehensive antioxidant activity Z value in this example is as follows: z is S1×0.6087+S2×0.1397+S3X 0.2516, and the combined antioxidant activities Z are listed in Table 2.
The combined antioxidant activities Z are also listed in Table 4.
TABLE 4
Figure BDA0003083381990000125
Figure BDA0003083381990000131
Taking Z as an effect value, and optimizing the extraction process of the water-soluble antioxidant ingredients of the ganoderma lucidum mycelia by using Design-Expert, wherein the optimized process comprises the following steps: the liquid-solid ratio is 50mL/g, the ultrasonic time is 60min, the ultrasonic temperature is 50 ℃, the ultrasonic power is 156W, the predicted value of the comprehensive antioxidant activity Z under the process condition is 52.64%, and the experimental value obtained by the verification experiment is 52.55%, and the two are similar (RSD is less than 3%). The corresponding radical clearance rates are in order: 38.68% (DPPH radical clearance), 53.79% (hydroxyl radical clearance) and 65.37% (ABTS radical clearance).
The comprehensive antioxidant activity Z is taken as an effect value to obtain higher DPPH free radical clearance rate, hydroxyl free radical clearance rate and ABTS free radical clearance rate of the aqueous extract under the optimized extraction condition.

Claims (9)

1. A method for comprehensively evaluating the antioxidant activity of ganoderma lucidum mycelia is characterized by comprising the following steps: the method comprises the following steps:
(1) preparing water extract from ganoderma lucidum mycelia;
(2) respectively measuring various antioxidant evaluation index values of the water extract;
(3) weighting the antioxidant evaluation index value in the step (2) by an entropy weight method;
(4) and (3) calculating to obtain a comprehensive antioxidant activity Z value of the ganoderma lucidum mycelia according to the antioxidant evaluation index value in the step (2) and the weight obtained in the step (3).
2. The method according to claim 1, wherein in step (2), the antioxidant evaluation index value is DPPH radical clearance, hydroxyl radical clearance and/or ABTS radical clearance.
3. The method according to claim 1 or 2, wherein in the step (1), the aqueous extract is prepared under the conditions: precisely weighing 0.05-2 g of ganoderma lucidum mycelium solid, carrying out ultrasonic water extraction under the conditions of liquid-solid ratio of 20-70 mL/g, ultrasonic temperature of 30-60 ℃, ultrasonic time of 15-75 min and ultrasonic power of 72-180W, centrifuging the solution after extraction, and taking a proper amount of supernatant to prepare 1-2 mg/mL of ganoderma lucidum mycelium extracting solution.
4. The method according to claim 2, wherein the DPPH radical clearance is determined in step (2) by: placing 2mL of the water extract in a brown volumetric flask, adding 2mL of DPPH free radical working solution with the absorbance of 0.7 +/-0.01, reacting for 30min in a dark place, measuring the absorbance at the position of 517nm, and adjusting to zero by using 95% ethanol;
then, DPPH radical clearance S was calculated according to equation (1)1Repeating the steps for three times, and taking an average value:
Figure FDA0003083381980000011
wherein A isiIs the absorbance, A, of the water extract after reaction with the DPPH free radical working solutionjThe background absorbance of the aqueous extract, measured using 95% ethanol instead of the DPPH free radical working solution, A0The blank absorbance was measured by replacing the aqueous extract with deionized water.
5. The method according to claim 2, wherein the hydroxyl radical scavenging rate of the aqueous extract in step (2) is measured by: putting 1mL of the water extract into a test tube with a plug, and sequentially adding 1mL of 9mmol/L FeSO4And 8.8mmol/L H2O2Standing the solution at room temperature for 10min, transferring into 1mL of 9mmol/L salicylic acid-ethanol solution, mixing, keeping temperature in 37 deg.C water bath for 30min, centrifuging after the water extract and salicylic acid reaction system react sufficiently, and detecting absorbance A of supernatant at 510nmi' zeroing with deionized water;
then, the hydroxyl radical clearance S was calculated according to the formula (2)2Repeating the steps for three times, and taking an average value:
Figure FDA0003083381980000021
wherein A isj' is to replace H with deionized water2O2Measuring the background absorbance of the water extract, A0' is the blank absorbance measured after the deionized water is used for replacing the water extract;
the salicylic acid reaction system refers to the reaction system H2O2With FeSO4The hydroxyl radical generated by the reaction is captured by salicylic acid to generate 2, 3-dihydroxybenzoic acid which has obvious absorption at 510 nm.
6. The method according to claim 2, wherein in step (2), the ABTS free radical clearance of the aqueous extract is determined by: adding 1mL of the water extract into a brown volumetric flask, adding 4mL of ABTS free radical working solution with the absorbance near 0.7, shaking up, reacting for 6min in a dark place, and measuring the absorbance of the solution at 734 nm;
then, ABTS radical clearance S was calculated according to equation (3)3The experiment was repeated three times, and the mean value was taken:
Figure FDA0003083381980000022
wherein A isi"is the absorbance, A, of the water extract after reaction with the ABTS free radical working solutionj"is the background absorbance of the sample solution measured after replacing the ABTS free radical working solution with phosphate buffer solution, A0"is the blank absorbance measured after the deionized water is used to replace the water extract.
7. The method according to claim 1, wherein in step (3), the weight is given by:
first, the antioxidant ability s of the water extract according to the formula (4)ijCarrying out dispersion standardization calculation to obtain yij；
Figure FDA0003083381980000023
Wherein s isijThe j-th antioxidant evaluation index value of the i-th extracting solution is 1, 2, 3, …, m, j is 1, 2, 3, …, n; y isijIs s isijValues normalized by dispersion;
then, the information entropy E is calculated as equation (5)j；
Figure FDA0003083381980000024
Finally, the weight w is calculated according to equation (6)j：
Figure FDA0003083381980000025
Wherein the content of the first and second substances,
Figure FDA0003083381980000026
is the proportion of the result of the ith group of experimental data under the j-th antioxidant evaluation index value, and when p isijWhen 0, define
Figure FDA0003083381980000031
yijIs s in formula (4)ijObtaining a numerical value after dispersion standardization; kappa is the antioxidant evaluation index value; m is the number of experimental data sets; lnm, lnpijRespectively represent m and pijNatural logarithm of (E)jThe information entropy of the j-th antioxidant evaluation index value is shown, and wj is the weight of the j-th antioxidant evaluation index value.
8. The method according to claim 1 or 2, wherein in step (4), the comprehensive antioxidant activity Z value is calculated by: z is S1×w1+S2×w2+S3×w3(ii) a Wherein S is1Is DPPH radical clearance, S2Is hydroxy radical scavenging rate, S3For ABTS radical clearance, w1、w2、w3Are respectively S1、S2、S3The corresponding weight.
9. The method as claimed in claim 3 or 5, wherein the centrifugation conditions are centrifugation at 6000-12000rpm for 3-15 min.
CN202110575404.3A 2021-05-25 2021-05-25 Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia Pending CN113298381A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110575404.3A CN113298381A (en) 2021-05-25 2021-05-25 Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110575404.3A CN113298381A (en) 2021-05-25 2021-05-25 Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia

Publications (1)

Publication Number Publication Date
CN113298381A true CN113298381A (en) 2021-08-24

Family

ID=77325023

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110575404.3A Pending CN113298381A (en) 2021-05-25 2021-05-25 Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia

Country Status (1)

Country Link
CN (1) CN113298381A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116415800A (en) * 2023-06-12 2023-07-11 山东瑞芝生物科技股份有限公司 Lucid ganoderma processing management system based on data model

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116415800A (en) * 2023-06-12 2023-07-11 山东瑞芝生物科技股份有限公司 Lucid ganoderma processing management system based on data model

Similar Documents

Publication Publication Date Title
Zhang et al. Polysaccharide and crocin contents, and antioxidant activity of saffron from different origins
Chen Optimization of extraction of Tremella fuciformis polysaccharides and its antioxidant and antitumour activities in vitro
Zhuang et al. Antioxidant capacity changes of 3 cultivar Chinese pomegranate (Punica granatum L.) juices and corresponding wines
CN111454844B (en) Novel ganoderma lucidum strain, ganoderma lucidum polysaccharide prepared based on ganoderma lucidum strain and anti-aging cosmetic
Ma et al. Purification, characterization, antioxidant activity and anti-aging of exopolysaccharides by Flammulina velutipes SF-06
Jing et al. Optimization of the extraction of polysaccharides from tobacco waste and their biological activities
CN108530549B (en) Method for extracting polysaccharide from ganoderma lucidum mycelia
Obluchinskaya Comparative chemical composition of the Barents Sea brown algae
CN112868850A (en) Preparation method of lucid ganoderma sporocarp drinking tablet tea capable of improving polysaccharide dissolution rate
CN113298381A (en) Method for comprehensively evaluating antioxidant activity of ganoderma lucidum mycelia
CN113499287A (en) Potato fermentation product, skin external preparation containing potato fermentation product, and preparation method and application of potato fermentation product
CN109400741A (en) A kind of isolation and purification method of ganoderma spore polysaccharide
CN108192931B (en) Method for preparing black garlic polyphenol by extracting black garlic through compound enzyme method
CN103585497A (en) Method for extracting and detecting polyphenol in bamboo leaves
CN112691125B (en) Pharmaceutical composition for whitening or resisting aging, preparation method thereof and skin care product
CN106117381A (en) A kind of method utilizing pectase to extract Polysaccharide from Portulaca oleracea
CN109912726B (en) Phellinus linteus polysaccharide derivative, preparation method and application thereof
CN114533628B (en) Red tassel seed sorghum cereal fermentation product, external skin preparation containing same and preparation method and application thereof
CN116334146A (en) Black tea double yeast fermentation product, cosmetic, and preparation methods and applications thereof
Nahas et al. Myco-antioxidants: insights into the natural metabolic treasure and their biological effects
CN113651898A (en) Coprinus comatus mycelium polysaccharide and preparation method and application thereof
CN110698567B (en) Antioxidant polysaccharide extract in pleurotus ferulae as well as preparation method and application thereof
Chioza et al. A comparative study on chemical composition and pharmacological effects of Paecilomyces hepiali and wild Ophiocordyceps sinensis
CN114747423A (en) Inonotus obliquus culture method and culture product thereof
CN112618427A (en) Kiwi fruit fermented juice and application thereof for improving skin condition

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination