LU503471B1 - Extraction method of proanthocyanidins from buckwheat fulls - Google Patents
Extraction method of proanthocyanidins from buckwheat fulls Download PDFInfo
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- LU503471B1 LU503471B1 LU503471A LU503471A LU503471B1 LU 503471 B1 LU503471 B1 LU 503471B1 LU 503471 A LU503471 A LU 503471A LU 503471 A LU503471 A LU 503471A LU 503471 B1 LU503471 B1 LU 503471B1
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0096—Purification; Precipitation; Filtration
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
- C12P17/162—Heterorings having oxygen atoms as the only ring heteroatoms, e.g. Lasalocid
Abstract
The invention relates to the technical field of proanthocyanidins extraction, in particular to an extraction method of proanthocyanidins from buckwheat fulls, which specifically comprises the following steps: taking buckwheat hulls as raw materials, using ethanol-water solution as extraction solvent, performing ultrasonic extraction under heating conditions, and centrifuging to obtain proanthocyanidins extraction solution. By extracting proanthocyanidins with excellent antioxidant effect from buckwheat hulls, the invention improves the utilization value of buckwheat hulls, makes buckwheat hulls become products with higher production added value, and provides theoretical support for the deep processing and utilization of buckwheat hulls. Experiments prove that the extraction amount of proanthocyanidins in buckwheat hulls is as high as 35.22mg/g, the free radical scavenging rate is as high as 94.64%, and the average polymerization degree is as low as 2.5.
Description
DESCRIPTION LU503471
EXTRACTION METHOD OF PROANTHOCYANIDINS FROM BUCKWHEAT
FULLS
The invention relates to the technical field of proanthocyanidins extraction, in particular to an extraction method of proanthocyanidins from buckwheat fulls.
Proanthocyanidins are a kind of polyphenol compounds polymerized by different kinds and quantities of flavan -3- alcohols through carbon-carbon (C-C) bonds. These substances can produce red proanthocyanidins under acidic heating conditions, and are internationally recognized as effective natural antioxidants for scavenging free radicals in human bodies. With the deepening of research, it is also found that they have high medicinal values such as antibacterial, anti-inflammatory and tumor inhibition.
Buckwheat belongs to dicotyledonous Polygonaceae, with high yield in China, ranking second in the world and abundant resources, which are mainly divided into
Fagopyrum esculentum and Fagopyrum tataricum. It is found that black bitter buckwheat hull is the highest content of polyphenol compounds such as proanthocyanidins.
However, at present, buckwheat hulls produced after black bitter buckwheat grain processing are not fully utilized, and some of them are made into buckwheat pillows, while the rest is discarded as garbage. Therefore, it is particularly necessary to provide an extraction method of proanthocyanidins from buckwheat fulls and make full and effective use of agricultural and sideline products.
SUMMARY LU503471
The invention aims to provide an extraction method of proanthocyanidins from buckwheat hulls, which makes full use of buckwheat hulls and improves the economic value of agricultural and sideline products.
One of the technical schemes of the invention is an extraction method of proanthocyanidins from buckwheat hulls, comprising: taking buckwheat hulls as raw materials and ethanol-water solution as extraction solvent, performing ultrasonic extraction under heating conditions, and centrifuging to obtain proanthocyanidins extract.
Water, ethanol and ethanol-water solution were used as extraction solvents to extract proanthocyanidins from buckwheat hull. It was found that ethanol-water solution was the highest extraction solvent.
Furthermore, the volume fraction of ethanol in the ethanol-water solution is 50-60%, the heating temperature is 30-70°C, the ultrasonic condition is 100-500W, the extraction time is 30-70min, and the ratio of buckwheat hull to ethanol-water solution is 1g: (10-30) ml.
Ultrasonic conditions are conducive to the dissolution of proanthocyanidins in solvents, and at the same time, ultrasonic can destroy the cell wall of buckwheat hull, promote the separation of proanthocyanidins and increase the extraction amount of proanthocyanidins. However, the heat generated by excessive power will destroy the structure of proanthocyanidins, thus reducing the extraction amount of proanthocyanidins. When the extraction temperature increased from 30°C to 50°C, the extraction amount of proanthocyanidins increased, and then began to decrease, because the increase of temperature would increase the kinetic energy of molecules, promote the dissolution of proanthocyanidins and increase the extraction amount of proanthocyanidins. When the extraction temperature continues to rise, the extraction amount of proanthocyanidins decreases, which may be due to the oxidation or degradation of proanthocyanidins at too high a temperature. When the extraction time increased from 20min to 30min, the extraction amount of proanthocyanidins increased with the increase of time. When the ultrasonic time was 30min to 50min, the extractidry503471 amount of proanthocyanidins increased slightly, but there was no obvious difference with other levels. When the ultrasonic time continued to increase, the extraction amount of proanthocyanidins decreased, which may be due to the structural damage of proanthocyanidins caused by long-term ultrasonic mechanical vibration, thus reducing the extraction amount of proanthocyanidins. The extraction amount of proanthocyanidins increased at first and then decreased slowly with the increase of the ratio of material to liquid. When the solvent amount is small, the extraction of proanthocyanidins is not sufficient. When the ratio of material to liquid reaches 1: 20 (g/ml), the extraction amount of proanthocyanidins is the largest, and then the extraction amount of proanthocyanidins decreases slowly with the increase of solvent amount. It may be that with the increase of solvent, too many impurities dissolve, which hinders the dissolution of proanthocyanidins, reduces the extraction amount, causes solvent waste, and increases the cost.
Further, the buckwheat hull is subjected to cellulase hydrolysis before extracting proanthocyanidins.
Further, the method specifically comprises the following steps:
Pretreatment: crushing buckwheat hulls to obtain buckwheat hull powder;
Enzymolysis: placing buckwheat hull powder in a buffer solution with a pH value of 4.5-6, preheating, adding cellulase for enzymolysis, inactivating, and centrifuging the product to obtain supernatant a and substrate a;
Ethanol extraction: transferring substrate a into ethanol-water solution for heating and ultrasonic extraction, and centrifuging the product to obtain supernatant b and substrate b; combining supernatant a and supernatant b to obtain proanthocyanidin extract.
Further, in the enzymolysis step, the ratio of buckwheat hull powder to buffer solution is 1g: (10-15) ml, and the addition amount of cellulase is 5-10% of the mass of buckwheat hull; preheating at 45-60°C, enzymolysis at 45-60°C for 30-60min; Inactivation includes heating at 90-100°C for 5-15min.
Before the extraction of proanthocyanidins, cellulase can catalyze the hydrolysis of cellulose in buckwheat hull, break the cell wall, increase the permeability of the membrane, and promote the dissolution of intracellular substances. After tH&J503471 enzymolysis, it is helpful to protect proanthocyanidins from being damaged in the extraction process and improve the biological activity of the extracted proanthocyanidins.
Further, before extracting proanthocyanidins, the buckwheat hull is fermented by microbial agents, and the microbial agents include 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: crushing buckwheat hulls to obtain buckwheat hull powder;
Fermentation: after the buckwheat hull powder and microbial agent are evenly mixed according to the mass ratio of 100: (5-10), the water content is adjusted to 40-60%, and the sealed solid-state fermentation is carried out at 30-40°C for 1-2 days, and the product is dried and crushed to obtain the fermentation substrate;
Ethanol extraction: the fermentation substrate is transferred into ethanol-water solution for heating and ultrasonic extraction, and the product is isolated to obtain proanthocyanidin extract and precipitate.
Proanthocyanidins usually form stable molecular complexes with protein and polysaccharides by hydrogen bonds in plants, which is the key factor affecting the extraction efficiency of proanthocyanidins. Before extraction, pectinase, a metabolite produced by Paenibacillus polymyxa fermentation, can fully decompose macromolecular compounds such as pectin in plant cell tissues, destroy cell walls and reduce the resistance of extraction mass transfer. Combining with Bacillus subtilis to synthesize a-amylase, protease, lipase, cellulase and other enzymes, fully destroy the structure of buckwheat hull cells, changing the material morphology, reducing the hydrogen bond stability of proanthocyanidins, protein and polysaccharide molecular complex, and reducing the polymerization degree of proanthocyanidins. Yeast contains ethanol, which can ensure the stable release of proanthocyanidins released by Paenibacillus polymyxa and Bacillus subtilis, thus contributing to the rapid release of proanthocyanidins in the subsequent extraction process.
Further, a pulsed electric field is applied in the extraction process;
Further, the pulsed electric field conditions are: electric field intensity of 10-20kv/crhU503471 discharge times of 10-15 times, and time of applying pulsed electric field for 5-10min.
Pulsed electric field will affect the stability of high-molecular-weight proanthocyanidins, which decomposes high-molecular-weight proanthocyanidins into low-molecular-weight proanthocyanidins, thus increasing the extraction amount of proanthocyanidins and enhancing their biological activity.
The second technical scheme of the invention is that the proanthocyanidins extracted by the method for extracting proanthocyanidins from buckwheat hulls.
Compared with the prior art, the invention has the advantages that:
The invention provides an extraction method of proanthocyanidins from buckwheat hulls. By extracting proanthocyanidins with excellent antioxidant effect from buckwheat hulls, the utilization value of buckwheat hulls is improved, so that buckwheat hulls become products with higher production added value, and theoretical support is provided for the deep processing and utilization of buckwheat hulls. Experiments prove that the extraction amount of proanthocyanidins in buckwheat hulls is as high as 35.22mg/g, the free radical scavenging rate is as high as 94.64%, and the average polymerization degree is as low as 2.5.
Fig. 1 is a standard curve of proanthocyanidins content in an embodiment of the present invention;
Fig. 2 is a standard curve of the polymerization degree of proanthocyanidins in an embodiment of the present invention.
DESCRIPTION OF THE INVENTION LU503471
Now, various exemplary embodiments of the present invention will be described in detail. This detailed description should not be taken as a limitation of the present invention, but should be understood as a more detailed description of some aspects, characteristics and embodiments of the present invention.
It should be understood that the terms mentioned in the present invention are only used to describe specific embodiments, and are not used to limit the present invention.
In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Every smaller range between any stated value or the intermediate value within the stated range and any other stated value or the intermediate value within the stated range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.
Unless otherwise stated, all technical and scientific terms used herein have the same meanings commonly understood by those of ordinary skill in the field to which this invention relates. Although the present invention only describes preferred methods and materials, 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 to disclose and describe the methods and/or materials related to the documents. In case of conflict with any incorporated documents, the contents of this specification shall prevail.
Without departing from the scope or spirit of the present invention, it is obvious to those skilled in the art that many modifications and changes can be made to the specific embodiments of the present specification. Other embodiments obtained from the description of the present invention will be obvious to the skilled person. The specification and embodiment of this application are only exemplary.
As used in this paper, the terms "comprising", "including", "having" and "containing" are all open terms, meaning including but not limited to.
In the following examples of the invention, black bitter buckwheat hulls are used d44J503471 raw materials of buckwheat hulls, which are crushed by a hammer mill, screened by a 0.3mm sieve, sealed by an ethylene bag and stored at 4°C. The main material components are shown in Table 1.
Table 1
In the following examples of the present invention, the content of proanthocyanidins is measured by:
Vanillin-hydrochloric acid method: accurately prepare 1mg/ML mother liquor of catechin standard, and dilute the mother liquor to obtain standard solutions with concentrations of 0.4mg/mL, 0.3mg/mL, 0.2mg/mL, 0.1mg/ML and 0.05mg/mL. Add 3mL of methanol solution containing 4% Vanillin to 1ml of standard solution, shake well, then add 1.5mL of concentrated hydrochloric acid, and react at 25°C in the dark for 15min.
Measure the absorbance at 500 nm, and replace the sample with methanol as the blank group. Specify the standard curve of proanthocyanidins mass concentration, as shown in
Figure 1;
Evaporate the proanthocyanidins extract under reduced pressure to dryness, fix the volume with methanol, put 1 mL into a test tube with a plug, and detect its absorbance at 500nm by the above method. The mass concentration (mg /mL) of proanthocyanidins was obtained by reference curve, and the yield of proanthocyanidins was calculated according to the following formula.
Yield of proanthocyanidins (%) = x 100% , where C is the mass concentration of proanthocyanidins in the measured solution, mg /mL; N is the dilution multiple of the extractive solution; V is the total volume of the extract, ml; M is the mass of the sample powder, g.
In the following examples of the present invention, the method for measuring the polymerization degree of proanthocyanidins:
Improved Vanillin-hydrochloric acid method: accurately prepare 1umol /mL acetic acid mother liquor of catechin standard, and dilute the mother liquor to obtain standard solutions with concentrations of 0.03umol/mL, 0.06 jumol/ML, 0.09 pmol/mL, 011503471 pmol/mL and 0.15 umol/mL.
Add 5 mL of 0.5% Vanillin-acetic acid solution containing 4% hydrochloric acid to 1 ml of sample, react at 20°C in the dark for 20 min, measure the absorbance at 500 nm, and take acetic acid instead of the sample as the blank group.
Establish the standard curve of proanthocyanidins molar concentration, as shown in
Figure 2;
Evaporate the proanthocyanidins extract under reduced pressure to dryness, fix the volume with acetic acid, put 1 mL into a test tube with a plug, and detect its absorbance at 500nm by the above method. The molar concentration of proanthocyanidins (umol/mL) was obtained by reference curve, and the average polymerization degree of proanthocyanidins was calculated according to the following formula.
Average degree of polymerization (mDP)= — ,
In the formula, m represents the mass of proanthocyanidins (mg), n represents the amount of proanthocyanidins (mmoL), and m represents the molar mass of proanthocyanidins (catechins) (270 g/MOL).
Example 1
Weigh 1 g of black bitter buckwheat hull powder into a 50 mL test tube with a plug, add 60% ethanol solution according to a certain ratio of material to liquid (g/ml), extract by temperature and ultrasound, centrifuge at 4000 r/min for 10 min, and take the supernatant to determine the content of proanthocyanidins.
The single factor experiment was used to investigate the effects of solid-liquid ratio, temperature and ultrasonic conditions on the extraction of proanthocyanidins, as follows: (1) the extraction temperature is 50°C, the extraction time is 30 min, the ratio of solid to liquid is 1:20(g:ml), and the ultrasonic power is 100W, 200W, 300W, 400W and 500W respectively;
The results showed that when the ultrasonic power increased from 100W to 500W, the extraction amount of proanthocyanidins increased at first and then decreased. When the ultrasonic power reached 300W the extraction amount of proanthocyanidins reached the maximum value, which was significantly higher than that of other levels. This is because with the increase of ultrasonic power, the damage of ultrasonic to the cell wall bE503471 buckwheat hull is enhanced, which promotes the separation of proanthocyanidins and increases the extraction amount of proanthocyanidins. However, the heat generated by excessive power will destroy the structure of proanthocyanidins, thus reducing the extraction amount of proanthocyanidins. (2) Fixed extraction time 30min, solid-liquid ratio of 1:20(g:ml), ultrasonic power of 300W, extraction temperatures of 30°C, 40°C, 50°C, 60°C and 70°C respectively;
The results showed that when the extraction temperature increased from 30°C to 50°C, the extraction amount of proanthocyanidins increased, and then began to decrease, because the increase of temperature would increase the kinetic energy of molecules, promote the dissolution of proanthocyanidins and increase the extraction amount of proanthocyanidins. When the extraction temperature continues to rise, the extraction amount of proanthocyanidins decreases, which may be due to the oxidation or degradation of proanthocyanidins at too high a temperature. (3) The solid-liquid ratio is 1:20(g:ml), the ultrasonic power is 300W, the extraction temperature is 50°C, and the extraction time is 20min, 30min, 40min, 50min and 60min, respectively;
The results showed that when the extraction time increased from 20 min to 30min, the extraction amount of proanthocyanidins increased with the increase of time. When the ultrasonic time was from 30 min to 50 min, the extraction amount of proanthocyanidins increased slightly, but there was no obvious difference with other levels. When the ultrasonic time continued to increase, the extraction amount of proanthocyanidins decreased, which may be due to the structural damage of proanthocyanidins caused by long-term ultrasonic mechanical vibration, thus reducing the extraction amount of proanthocyanidins. (4) The ultrasonic power was 300W, the extraction temperature was 50°C, the extraction time was 30min, and the ratio of solid to liquid was 1:10, 1:15, 1:20, 1:25 and 1:30, respectively.
The results showed that the extraction amount of proanthocyanidins increased at first and then decreased slowly with the increase of the ratio of material to liquid. It shows that the extraction of proanthocyanidins is not sufficient when the solvent amount l$/503471 small. When the ratio of material to liquid reaches 1: 20 (g/ml), the extraction amount of proanthocyanidins is the largest, and then the extraction amount of proanthocyanidins decreases slowly with the increase of solvent amount, which may be due to the excessive impurities dissolved with the increase of solvent, which hinders the dissolution of proanthocyanidins, reduces the extraction rate, and causes solvent waste and increases the cost.
Combined with the above contents, the response surface optimization results were used to analyze and verify the optimum extraction process parameters of proanthocyanidins. The optimum extraction process parameters of proanthocyanidins from black tartary buckwheat were as follows: ultrasonic power 287W, ultrasonic temperature 46°C, ultrasonic time 33min, and solid-liquid ratio 1:18(g:ml). Under these conditions, three parallel experiments were conducted, and the extraction amount of proanthocyanidins was 25.16 mg/g.
Example 2 (1) Buckwheat hull powder is put in a phosphate buffer solution with a pH value of 5.5 (the ratio of material to liquid is 1g:10mL), preheated at 55°C, then added with cellulase (5% of the weight of buckwheat hull) for enzymolysis at 55°C for 40min, inactivated at 95°C for 15min, and the product is centrifuged to obtain supernatant a and substrate a; (2) Ethanol extraction: substrate a is transferred into 60% ethanol-water solution according to the ratio of material to liquid 1g: 18ml, and then heated at 46°C for 287W for ultrasonic extraction for 33min, and the product is centrifuged to obtain supernatant b and substrate b; combining supernatant a and supernatant b to obtain proanthocyanidin extract; the extraction amount of proanthocyanidins was 32.05mg/g; (3) One of the following conditions was changed by single factor experiment: pH value of phosphate buffer solution was 4.5, 5 and 6, the ratio of solid to liquid was 19:15mL, and the enzymolysis temperature was 45°C, 50°C and 60°C respectively. The results showed that the extraction amount of proanthocyanidins was in the range of 28.68-30.85mg/g.
Example 3 LU503471
As in Example 2, the difference is that the inactivation step of cellulase is omitted, specifically: (1) buckwheat hull powder is placed in a phosphate buffer solution with a pH value of 5.5 (the ratio of material to liquid is 1g:10mL), preheated at 55°C, and then added with cellulase (5% of the weight of buckwheat hull) for enzymolysis at 55°C for 40min, and the product is centrifuged to obtain supernatant a and substrate a; (2) ethanol extraction: substrate a is transferred into 60% ethanol-water solution according to the ratio of material to liquid 1g: 18ml, and then heated at 46°C for 287W for ultrasonic extraction for 33min, and the product is centrifuged to obtain supernatant b and substrate b; combining supernatant a and supernatant b to obtain proanthocyanidin extract; the extraction amount of proanthocyanidins was 26.75mg/g after detection.
Example 4 (1) commercial yeast, Paenibacillus polymyxa and Bacillus subtilis are mixed according to the mass ratio of 5:4:3 to obtain microbial agent; (2) after buckwheat hull powder and microbial agent are evenly mixed according to the mass ratio of 100:8, water is added to adjust the water content to 50%, and the mixture is transferred to a sealed environment at 35°C for fermentation for 2 days, and the product is dried and crushed at 50°C to obtain fermentation substrate; (3) according to the ratio of material to liquid 1g:18ml, the fermentation substrate was transferred into 60% ethanol-water solution, heated at 46°C for 287W for ultrasonic extraction for 33min, and the product was isolated to obtain supernatant b and substrate b; combining supernatant a and supernatant b to obtain proanthocyanidin extract; The extraction amount of proanthocyanidins was 33.46 mg/g; (4) one of the following conditions was changed by single factor experiment: the mass ratio of yeast, Paenibacillus polymyxa and Bacillus subtilis was 10:2:1, 5:5:1, 8:4:3, 5:4:2, and the mass ratio of buckwheat hull powder and microbial agent was 100:5, 100: 10. The results showed that the extraction amount of proanthocyanidins was in the range of 30.15-32.49 mg/g.
Example 5
(1) commercial yeast, Paenibacillus polymyxa and Bacillus subtilis are mixe&d/503471 according to the mass ratio of 5:4:3 to obtain microbial agent; (2) after buckwheat hull powder and microbial agent are evenly mixed according to the mass ratio of 100:8, water is added to adjust the water content to 50%, and the mixture is transferred to a sealed environment at 35°C for fermentation for 2 days, and the product is dried and crushed at 50°C to obtain fermentation substrate; (3) according to the ratio of material to liquid 1g: 18ml, the fermentation substrate was transferred into 60% ethanol-water solution, and then heated at 46°C for 287W for ultrasonic extraction for 8min. Eight minutes before the end of ultrasonic hydrolysis, a pulsed electric field (electric field intensity 15 kv/cm, discharge frequency 15 times) was applied, and the product was centrifuged to obtain supernatant b and substrate b; combining supernatant a and supernatant b to obtain proanthocyanidin extract; the extraction amount of proanthocyanidins was 35.22mg/g.
DPPH method was used to determine the free radical scavenging rate of the proanthocyanidin extract obtained under the best conditions in Example 1 (ultrasonic power 287W, ultrasonic temperature 46 °C, ultrasonic time 33 min, material to liquid ratio 1:18 (g: ml), the proanthocyanidin extract prepared in (2) of Example 2, the proanthocyanidin extract prepared in (2) of Example 3, the proanthocyanidin extract prepared in (3) of Example 4, and the proanthocyanidin extract prepared in (3) of
Example 5. See Table 2 for the results;
The average degree of polymerization degree of proanthocyanidins in the best conditions of Example 1 (Ultrasonic power 28/W, ultrasonic temperature 46 °C, ultrasonic time 33min, material liquid ratio 1:18 (g: ml)), the proanthocyanidin extract prepared in (2) of Example 2, the proanthocyanidin extract prepared in (2) of Example 3, the proanthocyanidin extract prepared in (3) of Example 4, and the proanthocyanidin extract prepared in (3) of Example 5 were determined by ultraviolet spectrophotometry.
See Table 2 for the results;
Table 2
Free radical scavenging | Average degree of
I ial rl
The above is only a preferred embodiment of the present invention, and it is not intended to limit the present invention. Any modification, equivalent substitution and improvement made within the spirit and principle of the present invention should be included in the scope of protection of the present invention.
Claims (10)
1. An extraction method of proanthocyanidins from buckwheat hulls, characterized by comprising: using buckwheat hulls as raw materials and ethanol-water solution as extraction solvent, performing ultrasonic extraction under heating conditions, and centrifuging to obtain proanthocyanidins extract.
2. The extraction method of proanthocyanidins from buckwheat hulls according to claim 1, characterized in that the volume fraction of ethanol in the ethanol-water solution is 50-60%, the heating temperature is 30-70°C, the ultrasonic condition is 100-500W, the extraction time is 30-70min, and the ratio of buckwheat hulls to ethanol-water solution is 1g: (10-30) ml.
3. The extraction method of proanthocyanidins from buckwheat hulls according to claim 1, characterized in that the buckwheat hull is subjected to cellulase hydrolysis before extracting proanthocyanidins.
4. The extraction method of proanthocyanidins from buckwheat hulls according to claim 3, characterized by comprising pretreatment: crushing buckwheat hulls to obtain buckwheat hull powder; enzymolysis: placing buckwheat hull powder in a buffer solution with a pH value of
4.5-6, preheating, adding cellulase for enzymolysis, inactivating, and centrifuging the product to obtain supernatant a and substrate a; ethanol extraction: transferring substrate a into ethanol-water solution for heating and ultrasonic extraction, and centrifuging the product to obtain supernatant b and substrate b; combining supernatant a and supernatant b to obtain proanthocyanidin extract.
5. The extraction method of proanthocyanidins from buckwheat hulls according tà/503471 claim 4, characterized in that in the enzymolysis, the material-liquid ratio of buckwheat hull powder to buffer solution is 1g: (10-15) ml, and the addition amount of cellulase is 5-10% of the mass of buckwheat hull; preheating at 45-60°C, enzymolysis at 45-60°C for 30-60min; inactivation at 90-100°C for 5-15min.
6. The extraction method of proanthocyanidins from buckwheat hulls according to claim 1, characterized in that before extracting proanthocyanidins, buckwheat hulls are fermented by microbial agents, and the microbial agents include 5-10 parts of yeast, 2-5 parts of Paenibacillus polymyxa and 1-3 parts of Bacillus subtilis.
7. The extraction method of proanthocyanidins from buckwheat hulls according to claim 6, characterized by comprsing pretreatment: crushing buckwheat hulls to obtain buckwheat hull powder; fermentation: after mixing the buckwheat hull powder and microbial agent evenly according to the mass ratio of 100: (5-10), adjusting the water content to 40-60%, and carrying out the sealed solid-state fermentation at 30-40°C for 1-2 days, and drying and crushing the product to obtain the fermentation substrate; ethanol extraction: transferring the fermentation substrate into ethanol-water solution for heating and ultrasonic extraction, and isolating the product to obtain proanthocyanidin extract and precipitate.
8. The extraction method of proanthocyanidins from buckwheat hulls according to claim 7, characterized in that in the extraction process, a pulsed electric field is applied.
9. The extraction method of proanthocyanidins from buckwheat hulls according to claim 8, characterized in that the pulsed electric field conditions are: the electric field intensity is 10-20kv/cm, the discharge times are 10-15 times, and the applied pulsed electric field time is 5-10min.
10. A proanthocyanidin extracted from buckwheat hulls according to the extractidrv503471 method of proanthocyanidin according to any one of claims 1-9.
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| CN113173903B (en) * | 2021-06-16 | 2022-03-25 | 河南工业大学 | Method for extracting procyanidine from buckwheat hulls |
| CN116210830A (en) * | 2023-02-08 | 2023-06-06 | 西北农林科技大学 | Blood sugar-reducing compound anthocyanin beverage and preparation method thereof |
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| CN103992300B (en) * | 2014-05-30 | 2015-12-02 | 云南朱提苦荞生物科技开发有限公司 | A kind of method extracting pycnogenols from tartary buckwheat bran |
| CN106187977A (en) * | 2016-07-06 | 2016-12-07 | 丘利锦 | A kind of method extracting Garcinia mangostana shell procyanidins |
| CN113173903B (en) * | 2021-06-16 | 2022-03-25 | 河南工业大学 | Method for extracting procyanidine from buckwheat hulls |
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| Publication number | Publication date |
|---|---|
| CN113173903B (en) | 2022-03-25 |
| WO2022262456A1 (en) | 2022-12-22 |
| CN113173903A (en) | 2021-07-27 |
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