CN113173903B - Method for extracting procyanidine from buckwheat hulls - Google Patents
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Abstract
The invention relates to the technical field of procyanidine extraction, in particular to a method for extracting procyanidine from buckwheat hulls, which specifically comprises the following steps: the method comprises the following steps of taking buckwheat hulls as raw materials, taking an ethanol water solution as an extraction solvent, carrying out ultrasonic extraction under a heating condition, and centrifuging to obtain a proanthocyanidins extracting solution; by extracting the procyanidine with excellent antioxidant effect from the buckwheat hulls, the utilization value of the buckwheat hulls is improved, the buckwheat hulls become products with higher production added value, and theoretical support is provided for deep processing and utilization of the buckwheat hulls. Tests prove that the extraction amount of the procyanidine in the buckwheat hulls by the method is 35.22mg/g, the free radical clearance rate is as high as 94.64%, and the average polymerization degree is as low as 2.5.
Description
Technical Field
The invention relates to the technical field of procyanidine extraction, and particularly relates to a method for extracting procyanidine from buckwheat hulls.
Background
Procyanidine is a polyphenol compound which is polymerized by flavan-3-ol of different types and quantities through a carbon-carbon (C-C) bond, the substance can generate red anthocyanin under the condition of acid heating, is a natural antioxidant which is internationally acknowledged at present and is effective in eliminating free radicals in human bodies, and has higher medicinal values of antibiosis, antiphlogosis, tumor inhibition and the like along with the deep research.
Buckwheat belongs to dicotyledonous Polygonaceae plants, has high yield in China, is the second place in the world, is rich in resources and mainly comprises wild buckwheat and tartary buckwheat. Researches find that the black tartary buckwheat hulls are the parts with the highest content of polyphenol compounds such as procyanidine, however, the buckwheat hulls generated after the black tartary buckwheat grains are processed are not fully utilized, one part of the buckwheat hulls are made into a buckwheat pillow, and the rest buckwheat hulls are discarded as garbage. Therefore, the method for extracting the procyanidine from the buckwheat hulls is provided, and the method is especially necessary for fully and effectively utilizing agricultural and sideline product resources.
Disclosure of Invention
The invention aims to provide a method for extracting procyanidine from buckwheat hulls, which makes full use of buckwheat hull resources and improves the economic value of agricultural and sideline products.
One of the technical schemes of the invention is a method for extracting procyanidine from buckwheat hulls, which comprises the following steps: the method comprises the steps of taking buckwheat hulls as raw materials, taking an ethanol water solution as an extraction solvent, carrying out ultrasonic extraction under a heating condition, and centrifuging to obtain a proanthocyanidins extracting solution.
The extraction of the buckwheat hull procyanidin is carried out by taking water, ethanol and an ethanol aqueous solution as extraction solvents, and the extraction amount of the ethanol aqueous solution as the extraction solvent is the highest.
Further, the volume fraction of ethanol in the ethanol water solution is 50-60%, the heating temperature is 30-70 ℃, the ultrasonic condition is 100-: (10-30) mL.
The ultrasonic condition is helpful for dissolving procyanidin in solvent, and the ultrasonic wave has destructive effect on buckwheat hull cell wall, promotes procyanidin separation, increases procyanidin extraction amount, but the heat generated by over-power can destroy procyanidin structure, thereby reducing procyanidin extraction amount. When the extraction temperature is increased from 30 ℃ to 50 ℃, the extraction amount of procyanidin is increased and then starts to decrease, because the increase of the temperature can increase the molecular kinetic energy, promote the dissolution of procyanidin and improve the extraction amount of procyanidin. When the extraction temperature is increased continuously, the extraction amount of the procyanidin is reduced, probably because the procyanidin is easy to be oxidized or degraded due to the overhigh temperature. When the extraction time is increased from 20min to 30min, the extraction amount of the procyanidin is in an increasing trend along with the increase of the time, when the ultrasonic time is 30min to 50min, the extraction amount of the procyanidin is slightly increased, but the difference with other levels is not obvious, when the ultrasonic time is continuously increased, the extraction amount of the procyanidin is in a decreasing trend, probably because the structure of the procyanidin is damaged due to long-time ultrasonic mechanical vibration, and the extraction amount of the procyanidin is reduced. The extraction amount of the procyanidine is increased and then slowly reduced along with the increase of the feed-liquid ratio. When the solvent amount is small, the extraction of procyanidine is not sufficient, and when the material-liquid ratio reaches 1: at 20 g/mL, the extraction amount of procyanidin is the largest, and then the extraction amount of procyanidin is slowly reduced as the solvent amount increases, probably because the dissolved impurities are too much as the solvent increases, the dissolution of procyanidin is hindered, the extraction amount is reduced, the solvent is wasted, and the cost is increased.
Further, the buckwheat hulls are subjected to cellulose enzymolysis before procyanidin extraction.
Further, the method specifically comprises the following steps:
pretreatment: pulverizing buckwheat hulls to obtain buckwheat hull powder;
enzymolysis: placing the buckwheat hull powder into a buffer solution with the pH value of 4.5-6, preheating, adding cellulase for enzymolysis reaction, inactivating, and centrifuging the product to obtain a supernatant a and a substrate a;
alcohol extraction: transferring the substrate a into an ethanol water solution for heating ultrasonic extraction, and centrifuging the product to obtain a supernatant b and a substrate b; and combining the supernatant a and the supernatant b to obtain the procyanidin extracting solution.
Further, in the step of enzymolysis: the feed-liquid ratio of the buckwheat hull powder to the buffer solution is 1g: (10-15) mL, wherein the addition amount of the cellulase is 5-10% of the buckwheat hull by mass; preheating at 45-60 deg.C, performing enzymolysis at 45-60 deg.C, and performing enzymolysis for 30-60 min; the inactivation specifically comprises the following steps: heating at 90-100 deg.C for 5-15 min.
The method is characterized in that cellulose enzymolysis treatment is carried out before procyanidin is extracted, the cellulose in buckwheat hulls can be catalyzed by the cellulose to be hydrolyzed, cell walls are partially broken, the permeability of membranes is increased, substances in cells are dissolved out, enzyme deactivation operation is carried out after enzymolysis reaction is finished, and then centrifugation is carried out, so that procyanidin is protected from being damaged in the extraction process, and the biological activity of the extracted procyanidin is improved.
Further, before procyanidine extraction, the buckwheat hulls are fermented by a microbial agent, wherein the microbial agent comprises the following components in percentage by mass: 5-10 parts of yeast, 2-5 parts of paenibacillus polymyxa and 1-3 parts of bacillus subtilis.
Further, the method specifically comprises the following steps:
pretreatment: pulverizing buckwheat hulls to obtain buckwheat hull powder;
fermentation: mixing buckwheat hull powder and a microbial agent according to a mass ratio of 100: (5-10), after uniformly mixing, regulating the water content to 40-60%, carrying out sealed solid state fermentation for 1-2d at the temperature of 30-40 ℃, and drying and crushing a product to obtain a fermentation substrate;
alcohol extraction: and transferring the fermentation substrate into an ethanol water solution for heating and ultrasonic extraction, and centrifuging the product to obtain procyanidine extracting solution and precipitate.
Anthocyanin usually forms stable molecular compound with protein, polysaccharide and the like in a plant body through hydrogen bonds, which is a key factor influencing the extraction efficiency of procyanidine, and before extraction, the anthocyanin can fully decompose macromolecular compounds such as pectin and the like in plant cell tissues through microbial fermentation treatment and pectinase which is a metabolite generated by fermentation of Paenibacillus polymyxa, so that cell walls are damaged, and the extraction mass transfer resistance is reduced; the enzyme such as alpha-amylase, protease, lipase, cellulase and the like is synthesized by combining the bacillus subtilis thallus, the cell structure of the buckwheat hulls is fully damaged, the substance form in the buckwheat hulls is changed, the hydrogen bond stability of anthocyanin, protein and polysaccharide molecular compounds is reduced, and the polymerization degree of procyanidine is reduced. The ethanol component is contained in the yeast fermentation, so that the stable release of the procyanidin component released based on the fermentation of the paenibacillus polymyxa and the bacillus subtilis can be ensured, and the rapid release of the procyanidin in the subsequent extraction process is facilitated.
Further, a pulse electric field is applied in the extraction process;
further, the pulsed electric field conditions: the electric field intensity is 10-20kv/cm, the discharge frequency is 10-15 times, and the time of externally applying a pulse electric field is 5-10 min.
The pulse electric field can influence the stability of the high polymeric proanthocyanidins, so that the high polymeric proanthocyanidins are decomposed into oligomeric proanthocyanidins, the extraction amount of the proanthocyanidins is increased, and the biological activity of the proanthocyanidins is higher.
According to the second technical scheme, the procyanidine is extracted by the method for extracting procyanidine from buckwheat hulls.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides an extraction method of procyanidine in buckwheat hulls, which improves the utilization value of the buckwheat hulls by extracting procyanidine with excellent antioxidant effect from the buckwheat hulls, enables the buckwheat hulls to become products with higher production added value, and provides theoretical support for deep processing and utilization of the buckwheat hulls. Tests prove that the extraction amount of the procyanidine in the buckwheat hulls by the method is 35.22mg/g, the free radical clearance rate is as high as 94.64%, and the average polymerization degree is as low as 2.5.
Drawings
FIG. 1 is a standard curve of procyanidin content in an example of the invention;
FIG. 2 is a standard curve of the degree of polymerization of procyanidins according to the examples of the invention.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
In the following examples of the present invention, black tartary buckwheat hulls were used as a raw material of buckwheat hulls, the buckwheat hulls were crushed by a hammer mill and sieved through a 0.3mm sieve, and the buckwheat hulls were sealed with a vinyl bag and stored at 4 ℃ with the main material components shown in table 1;
TABLE 1
Composition (I) | Moisture content | Crude fat | Protein | Carbohydrate compound |
Content (%) | 12.41 | 3.53 | 11.25 | 32.87 |
In the following embodiments of the present invention, the method for measuring the content of procyanidine comprises:
vanillin-hydrochloric acid method: accurately preparing 1mg/mL catechin standard mother liquor, and diluting the mother liquor to obtain standard solutions with concentrations of 0.4mg/mL, 0.3mg/mL, 0.2mg/mL, 0.1mg/mL and 0.05 mg/mL. Respectively adding 1mL of standard solution into 3mL of methanol solution containing 4% vanillin, shaking, adding 1.5mL of concentrated hydrochloric acid, reacting at 25 deg.C in the dark for 15min, measuring light absorption value at 500nm, and using methanol as blank group instead of sample. Specifying a procyanidine mass concentration standard curve as shown in figure 1;
evaporating procyanidin extractive solution under reduced pressure to dryness, adding methanol to desired volume, placing 1mL into a test tube with a stopper, and detecting absorbance at 500nm by the above method. And (5) obtaining the mass concentration (mg/mL) of the procyanidine by contrasting with the standard curve, and calculating the yield of the procyanidine according to the following formula.
In the formula: c is the mass concentration of procyanidine in the solution to be tested, mg/mL; n is the dilution multiple of the extracting solution; v is the total volume of the extracting solution, mL; m is the mass of the sample powder, g.
In the following embodiments of the present invention, the method for measuring the degree of polymerization of procyanidine comprises:
modified vanillin-hydrochloric acid method: accurately preparing 1 mu mol/mL catechin standard substance acetic acid mother liquor, and diluting the mother liquor to obtain standard solutions with the concentrations of 0.03 mu mol/mL, 0.06 mu mol/mL, 0.09 mu mol/mL, 0.1 mu mol/mL and 0.15 mu mol/mL.
Adding 1mL sample into 5mL 4% hydrochloric acid 0.5% vanillin acetic acid solution, reacting at 20 deg.C in dark for 20min, measuring absorbance at 500nm, and using acetic acid as sample blank.
Preparing a procyanidine molar concentration standard curve shown in figure 2;
the procyanidin extract was evaporated to dryness under reduced pressure and the volume was determined with acetic acid, 1mL was taken in a stoppered tube and the absorbance at 500nm was measured by the method described above. The molar concentration (mu mol/mL) of the procyanidin is obtained by comparing with a standard curve, and the average polymerization degree of the procyanidin is calculated according to the following formula.
Wherein M represents the mass (mg) of procyanidin, n represents the amount (mmoL) of procyanidin, and M represents the molar mass (270g/moL) of procyanidin monomer (catechin).
Example 1
Weighing 1g of black tartary buckwheat hull powder into a 50mL test tube with a plug, adding 60% ethanol solution according to a certain material-liquid ratio (g/mL), performing temperature and ultrasonic assisted extraction, centrifuging at 4000r/min for 10min, and taking supernatant to determine the content of procyanidine.
The method adopts a single-factor experiment to investigate the influence of the material-liquid ratio, the temperature and the ultrasonic condition on the extraction amount of the procyanidine, and specifically comprises the following steps:
(1) the extraction temperature is fixed at 50 ℃, the extraction time is 30min, the material-liquid ratio is 1:20(g: ml), and the ultrasonic power is respectively 100W, 200W, 300W, 400W and 500W;
the results show that: when the ultrasonic power is increased from 100W to 500W, the extraction amount of procyanidin is increased and then decreased. When the ultrasonic power reaches 300W, the extraction amount of the procyanidine reaches the maximum value and is obviously higher than that of other levels. When the ultrasonic power is increased, the damage effect of the ultrasonic waves on the cell walls of the black tartary buckwheat hulls is enhanced, the separation of the procyanidine is promoted, the extraction amount of the procyanidine is increased, but the structure of the procyanidine is damaged by heat generated by the excessive power, so that the extraction amount of the procyanidine is reduced.
(2) The extraction time is 30min, the material-liquid ratio is 1:20(g: ml), the ultrasonic power is 300W, and the extraction temperature is 30 ℃, 40 ℃, 50 ℃, 60 ℃ and 70 ℃ respectively;
the results show that: when the extraction temperature is increased from 30 ℃ to 50 ℃, the extraction amount of procyanidin is increased and then starts to decrease, because the increase of the temperature can increase the molecular kinetic energy, promote the dissolution of procyanidin and improve the extraction amount of procyanidin. When the extraction temperature is increased continuously, the extraction amount of the procyanidin is reduced, probably because the procyanidin is easy to be oxidized or degraded due to the overhigh temperature.
(3) Fixing the ratio of material to liquid (g: ml) at 1:20, ultrasonic power of 300W, extraction temperature of 50 deg.C, and extraction time of 20min, 30min, 40min, 50min, and 60min respectively;
the results show that: when the extraction time is increased from 20min to 30min, the extraction amount of the procyanidin is in an increasing trend along with the increase of the time, when the ultrasonic time is 30min to 50min, the extraction amount of the procyanidin is slightly increased, but the difference with other levels is not obvious, when the ultrasonic time is continuously increased, the extraction amount of the procyanidin is in a decreasing trend, probably because the structure of the procyanidin is damaged due to long-time ultrasonic mechanical vibration, and the extraction amount of the procyanidin is reduced.
(4) The ultrasonic power is 300W, the extraction temperature is 50 ℃, the extraction time is 30min, and the material-liquid ratio is 1:10, 1:15, 1:20, 1:25 and 1:30 respectively.
The results show that: the extraction amount of the procyanidine is increased and then slowly reduced along with the increase of the feed-liquid ratio. The method indicates that when the solvent amount is small, the extraction of the procyanidine is not sufficient, and when the material-liquid ratio reaches 1: at 20 g/mL, the extraction amount of procyanidin is the largest, and then the extraction amount of procyanidin is slowly reduced due to the increase of solvent amount, probably because the dissolved impurities are too much along with the increase of solvent, the dissolution of procyanidin is hindered, the extraction rate is reduced, the waste of solvent is caused, and the cost is increased.
By combining the above contents, analyzing the response surface optimization result and combining the optimal procyanidine extraction process parameters and verification, the optimal process parameters for extracting procyanidine from black tartary buckwheat are obtained as follows: the ultrasonic power 287W, the ultrasonic temperature 46 ℃, the ultrasonic time 33min and the material-liquid ratio 1:18(g: ml) are carried out under the condition, and the extraction amount of the procyanidine is 25.16 mg/g.
Example 2
(1) Placing buckwheat hull powder into phosphoric acid buffer solution with pH value of 5.5 (material-liquid ratio is 1g:10mL), preheating at 55 deg.C, adding cellulase (5% of buckwheat hull mass) for enzymolysis at 55 deg.C for 40min, inactivating at 95 deg.C for 15min, centrifuging to obtain supernatant a and substrate a;
(2) alcohol extraction: substrate a is prepared according to the feed-liquid ratio of 1g: transferring 18mL of the mixture into an ethanol water solution with the volume fraction of 60%, heating to 287W at 46 ℃, performing ultrasonic extraction for 33min, and centrifuging the product to obtain a supernatant b and a substrate b; mixing the supernatant a and the supernatant b to obtain procyanidin extract; detection shows that the extraction amount of the procyanidine is 32.05 mg/g.
(3) One of the following conditions is changed through a single-factor experiment, the pH values of phosphoric acid buffer solutions are respectively 4.5, 5 and 6, the material-liquid ratio is 1g:15mL, the enzymolysis reaction temperatures are respectively 45 ℃, 50 ℃ and 60 ℃, and the result shows that the extraction amount of the procyanidine is in the range of 28.68-30.85 mg/g.
Example 3
The difference from example 2 is that the cellulase inactivation step was omitted, specifically:
(1) placing buckwheat hull powder into phosphoric acid buffer solution with pH value of 5.5 (material-liquid ratio is 1g:10mL), preheating at 55 deg.C, adding cellulase (5% of buckwheat hull mass) for enzymolysis at 55 deg.C for 40min, and centrifuging to obtain supernatant a and substrate a;
(2) alcohol extraction: substrate a is prepared according to the feed-liquid ratio of 1g: transferring 18mL of the mixture into an ethanol water solution with the volume fraction of 60%, heating to 287W at 46 ℃, performing ultrasonic extraction for 33min, and centrifuging the product to obtain a supernatant b and a substrate b; mixing the supernatant a and the supernatant b to obtain procyanidin extract; detection shows that the extraction amount of the procyanidine is 26.75 mg/g.
Example 4
(1) Mixing commercially available saccharomycetes, paenibacillus polymyxa and bacillus subtilis according to the mass ratio of 5:4:3 to obtain a microbial agent;
(2) uniformly mixing the buckwheat hull powder and the microbial agent according to the mass ratio of 100:8, adding water to adjust the water content to 50%, fermenting for 2d in a sealed environment at 35 ℃, and drying and crushing the product at 50 ℃ to obtain a fermentation substrate;
(3) fermentation substrate is prepared according to the feed liquid ratio of 1g: transferring 18mL of the mixture into an ethanol water solution with the volume fraction of 60%, heating to 287W at 46 ℃, performing ultrasonic extraction for 33min, and centrifuging the product to obtain a supernatant b and a substrate b; mixing the supernatant a and the supernatant b to obtain procyanidin extract; detection shows that the extraction amount of the procyanidine is 33.46 mg/g;
(4) one of the following conditions is changed through a single-factor experiment, wherein the mixing mass ratio of the yeast, the paenibacillus polymyxa and the bacillus subtilis is 10:2:1, 5:5:1, 8:4:3 and 5:4:2, and the mixing mass ratio of the buckwheat hull powder and the microbial agent is 100:5 and 100: 10, the results show that the procyanidin extraction was in the range of 30.15-32.49 mg/g.
Example 5
(1) Mixing commercially available saccharomycetes, paenibacillus polymyxa and bacillus subtilis according to the mass ratio of 5:4:3 to obtain a microbial agent;
(2) uniformly mixing the buckwheat hull powder and the microbial agent according to the mass ratio of 100:8, adding water to adjust the water content to 50%, fermenting for 2d in a sealed environment at 35 ℃, and drying and crushing the product at 50 ℃ to obtain a fermentation substrate;
(3) fermentation substrate is prepared according to the feed liquid ratio of 1g: transferring 18mL of the mixture into an ethanol aqueous solution with the volume fraction of 60%, heating to 287W at 46 ℃, performing ultrasonic extraction for 33min, applying a pulse electric field (the electric field intensity is 15kv/cm, the discharge frequency is 15 times) 8min before the ultrasonic hydrolysis is finished, and centrifuging the product to obtain a supernatant b and a substrate b; mixing the supernatant a and the supernatant b to obtain procyanidin extract; detection shows that the extraction amount of procyanidine is 35.22 mg/g.
The radical scavenging rate of the proanthocyanidin extract obtained under the optimum conditions (ultrasonic power 287W, ultrasonic temperature 46 ℃, ultrasonic time 33min, material-to-liquid ratio 1:18(g: ml)) in example 1, the proanthocyanidin extract prepared in step (2) in example 2, the proanthocyanidin extract prepared in step (2) in example 3, the proanthocyanidin extract prepared in step (3) in example 4, and the proanthocyanidin extract prepared in step (3) in example 5 was measured by DPPH method, and the results are shown in table 2;
the average polymerization degree of procyanidin in the best conditions (ultrasonic power 287W, ultrasonic temperature 46 ℃, ultrasonic time 33min, material to liquid ratio 1:18(g: ml)) of example 1, the procyanidin extracted solution prepared in step (2) of example 2, the procyanidin extracted solution prepared in step (2) of example 3, the procyanidin extracted solution prepared in step (3) of example 4, and the procyanidin extracted solution prepared in step (3) of example 5 was measured by ultraviolet spectrophotometry, and the results are shown in table 2;
TABLE 2
Clearance of free radicals,% | Average degree of polymerization | |
Example 1 | 75.25 | 5.4 |
Example 2 | 84.74 | 4.5 |
Example 3 | 81.45 | 4.2 |
Example 4 | 85.28 | 3.5 |
Example 5 | 94.64 | 2.5 |
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included therein.
Claims (4)
1. A method for extracting procyanidine from buckwheat hulls is characterized by comprising the following steps: the method comprises the following steps of taking buckwheat hulls as raw materials, taking an ethanol water solution as an extraction solvent, carrying out ultrasonic extraction under a heating condition, and centrifuging to obtain a proanthocyanidins extracting solution;
before procyanidine extraction, the buckwheat hulls are fermented by a microbial agent, wherein the microbial agent comprises the following strains in parts by weight: 5-10 parts of yeast, 2-5 parts of paenibacillus polymyxa and 1-3 parts of bacillus subtilis.
2. The method for extracting procyanidins from buckwheat hulls as claimed in claim 1, which comprises the following steps:
pretreatment: pulverizing buckwheat hulls to obtain buckwheat hull powder;
fermentation: mixing buckwheat hull powder and a microbial agent according to a mass ratio of 100: (5-10), after uniformly mixing, regulating the water content to 40-60%, carrying out sealed solid state fermentation for 1-2d at the temperature of 30-40 ℃, and drying and crushing a product to obtain a fermentation substrate;
alcohol extraction: and transferring the fermentation substrate into an ethanol water solution for heating and ultrasonic extraction, and centrifuging the product to obtain procyanidine extracting solution and precipitate.
3. The method for extracting procyanidins from buckwheat hulls as claimed in claim 2, wherein a pulsed electric field is applied during the extraction process.
4. The method for extracting procyanidins from buckwheat hulls as claimed in claim 3, wherein the pulsed electric field conditions are as follows: the electric field intensity is 10-20kv/cm, the discharge frequency is 10-15 times, and the time of externally applying a pulse electric field is 5-10 min.
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Application Number | Priority Date | Filing Date | Title |
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CN202110666063.0A CN113173903B (en) | 2021-06-16 | 2021-06-16 | Method for extracting procyanidine from buckwheat hulls |
PCT/CN2022/091140 WO2022262456A1 (en) | 2021-06-16 | 2022-05-06 | Method for extracting proanthocyanidins from buckwheat hulls |
LU503471A LU503471B1 (en) | 2021-06-16 | 2022-05-06 | Extraction method of proanthocyanidins from buckwheat fulls |
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