CN113307893A - Preparation and purification method of high-purity grifola frondosa polysaccharide extract - Google Patents
Preparation and purification method of high-purity grifola frondosa polysaccharide extract Download PDFInfo
- Publication number
- CN113307893A CN113307893A CN202110782672.2A CN202110782672A CN113307893A CN 113307893 A CN113307893 A CN 113307893A CN 202110782672 A CN202110782672 A CN 202110782672A CN 113307893 A CN113307893 A CN 113307893A
- Authority
- CN
- China
- Prior art keywords
- grifola frondosa
- purity
- polysaccharide
- membrane
- ultrafiltration
- 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.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Abstract
The invention relates to a preparation and purification method of a high-purity grifola frondosa polysaccharide extract, which comprises the following steps of (1) adding water into a grifola frondosa sporophore raw material, extracting at a high temperature, and filtering to obtain an extracting solution; (2) performing ultrafiltration concentration on the grifola frondosa extract by using a molecular sieve interception membrane; (3) purifying the ultrafiltration concentrated solution again by using ion exchange chromatography resin to obtain high-purity grifola frondosa polysaccharide extracting solution; (4) and drying the high-purity grifola frondosa polysaccharide extracting solution to obtain the grifola frondosa polysaccharide. The invention has the advantages that the purity of the prepared grifola frondosa polysaccharide is higher than that of the polysaccharide prepared by the traditional process; effectively preventing active ingredients in the grifola frondosa extracting solution from being damaged; the process is simple and convenient, other reagents such as ethanol and the like are not needed, and the production efficiency is improved; is more suitable for industrialization, effectively solves the problems of environmental protection and safety in industrial production, and has good economic benefit and social benefit.
Description
Technical Field
The invention relates to the technical field of extraction and separation of edible fungus grifola frondosa polysaccharide, and particularly relates to a preparation and purification method of a high-purity grifola frondosa polysaccharide extract.
Background
Grifola frondosa (Grifola frondosa) belongs to the kingdom of fungi, Basidiomycota, Basidiomycetes, Polyporales, Polyporaceae, Polyporus, also known as Polyporus lactiflora, Castanea mollissima, Foliaceae, Pleurotus ostreatus, and is called "Maitake" in Japan, and belongs to a precious fungus for both food and medicine. The chemical components of the grifola frondosa mainly comprise polysaccharide, protein, lipid and the like, wherein the main active component is grifola frondosa polysaccharide which is a fungal polysaccharide mainly composed of beta-1, 6-and beta-1, 3-glucosidic bonds and has obvious physiological activity, and the grifola frondosa polysaccharide is mainly alkali-soluble water extract. Studies have shown that grifolan has multiple active functions, such as: improving immunity, resisting cancer, resisting virus, resisting inflammation, lowering blood sugar, lowering blood pressure, etc. The foreign scientists extract a substance grifola frondosa polysaccharide which can effectively resist cancers from grifola frondosa, and researches show that the anticancer activity of the polysaccharide is 16 times of that of ganoderma lucidum. Meanwhile, the grifola frondosa can obviously improve the treatment effect and reduce the toxic and side effects of radiotherapy and chemotherapy through the synergistic action of the grifola frondosa and the chemotherapy, and the medicinal value of the grifola frondosa is immeasurable. The separation and extraction methods of the grifola frondosa polysaccharide are more, and the methods comprise hot water, acid and alkali extraction methods, complex enzyme and ultrasonic extraction, and each method has advantages and disadvantages. Meanwhile, deproteinization of grifola frondosa crude polysaccharide mainly comprises the following steps: the macroporous adsorption resin method, the trichloroacetic acid-n-butanol method and the Sevage method are mainly used, besides the above commonly used protein removal methods, the methods used in the literature include a tannic acid method, an isoelectric point method, a hydrochloric acid method, a sodium chloride method, a calcium chloride method and the like, and the method used also needs to select a proper method and various combinations according to the properties of a sample and the types of proteins. At present, the method is commonly used as a complex enzyme-microwave extraction method, and the enzymolysis extraction method has the advantages of high polysaccharide extraction rate, mild reaction conditions and small damage to polysaccharide structures, but is not suitable for large-batch industrial production because the price of the enzyme is high and the production cost is relatively high. The ultrasonic extraction method and the microwave extraction method can improve the extraction rate of the polysaccharide and shorten the extraction time, but sometimes cause the components of the polysaccharide extract to be more complex, and bring certain difficulty to the subsequent separation and purification process.
Disclosure of Invention
In view of the above disadvantages, the present invention aims to provide a method for preparing and purifying a high-purity grifola frondosa polysaccharide extract, which can greatly simplify the preparation process of grifola frondosa polysaccharide, and can extract high-purity polysaccharide, thereby realizing industrialized extraction and application.
In order to achieve the above object, the present invention provides the following technical solutions:
(1) adding water 10-40 times the weight of Grifola frondosa fruiting body into the fruiting body, extracting at 80-105 deg.C for 1-4 times, each for 1-4 hr, filtering the extractive solution with 80-200 mesh sieve, and mixing the extractive solutions;
(2) ultrafiltering the Grifola frondosa extractive solution with molecular sieve cut-off membrane, with cut-off molecular weight of 10-80kD, working pressure of 0.3-1Mpa, temperature of 25-50 deg.C, stopping ultrafiltration when the volume of the permeate is 40-70% of the total volume, collecting the cut-off concentrated solution,
(3) purifying the ultrafiltration concentrated solution again by using ion exchange chromatography resin, concentrating and purifying the obtained intercepted concentrated solution by using macroporous adsorption resin at the flow rate of 1-4BV/h and 3-10 times of cylinder accumulated water, wherein the elution flow rate is 0.5-5BV/h, then adjusting the pH value range to 6.5-8, and performing deproteinization treatment by using a cation and anion exchange resin series resin column; the mass ratio of the anion exchange resin to the cation resin is 1:1-5, so as to obtain a high-purity grifola frondosa polysaccharide extracting solution;
(4) drying the obtained high-purity grifola frondosa polysaccharide extract, controlling the moisture content of finished product powder to be less than 15%, and obtaining dry powder, namely the grifola frondosa polysaccharide finished product.
Preferably, the drying of the ultrafiltration retentate comprises one or more of spray drying, freeze drying, vacuum drying or atmospheric drying.
Preferably, the microfiltration membrane material is one or more of a ceramic membrane, an organic polymer membrane and a metal membrane.
Preferably, the ultrafiltration membrane has a cut-off molecular weight range of 10-80KDa, and the ultrafiltration membrane material is one or more of a ceramic membrane, an organic polymer membrane and a metal membrane.
Preferably, the ion exchange resin comprises one or more of anion exchange resin, cation exchange resin and nonionic exchange resin
The Grifola frondosa polysaccharide obtained by the above extraction method has the effects of improving immunity, resisting cancer, virus, inflammation, blood sugar and blood pressure, etc.
Compared with the prior art, the invention has the following beneficial effects:
(1) according to the preparation method provided by the invention, after extraction is finished, a molecular weight ultrafiltration technology is adopted for extraction and concentration, and then ion exchange adsorption combination is carried out, so that the preparation method has the advantages of simplicity in operation, low energy consumption, short production period and capability of using a filter membrane for multiple times and preventing active ingredients in the grifola frondosa extracting solution from being damaged, and the result shows that the purity of the prepared grifola frondosa polysaccharide is far higher than that of the grifola frondosa polysaccharide prepared by the traditional process.
(2) The operation process does not involve ethanol, so that the production cost is saved, and the convenience and safety of production operation are improved.
(3) The method is suitable for industrialization, effectively solves the problems of environmental protection and safety in industrial production, and has good economic benefit and social benefit.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example 1:
(1) adding water 20 times the weight of the raw materials into the Grifola frondosa fruiting body raw material, extracting at 100 deg.C for 4 times, each time for 2 hr, filtering the extractive solution with 200 mesh sieve, and mixing the extractive solutions;
(2) ultrafiltering the Grifola frondosa extractive solution with molecular sieve cut-off membrane, with cut-off molecular weight of 40kD, working pressure of 1Mpa, temperature of 40 deg.C, stopping ultrafiltration when the volume of the permeate is 50% of the total volume, and collecting the cut-off concentrated solution;
(3) purifying the ultrafiltration concentrated solution again by using ion exchange chromatography resin, concentrating and purifying the obtained intercepted concentrated solution by using 10 times of column accumulated water at the flow rate of 4BV/h by using macroporous adsorption resin, wherein the elution flow rate is 2BV/h, then adjusting the pH value range to 7, and treating by using a cation and anion exchange resin series resin column; the mass ratio of the anion exchange resin to the cation resin is 1:1, so as to obtain a high-purity grifola frondosa polysaccharide extracting solution;
(4) spray drying the obtained high-purity grifola frondosa polysaccharide extract, and controlling the water content of finished product powder to be less than 15%, wherein the obtained dry powder is the grifola frondosa polysaccharide finished product.
Example 2:
(1) adding water 40 times the weight of the raw materials into the Grifola frondosa fruiting body raw material, extracting at 100 deg.C for 2 times, each time for 1 hr, filtering the extractive solution with 200 mesh sieve, and mixing the extractive solutions;
(2) ultrafiltering the Grifola frondosa extractive solution with molecular sieve cut-off membrane, with cut-off molecular weight of 10kD, working pressure of 0.5Mpa, temperature of 40 deg.C, stopping ultrafiltration when the volume of the permeate is 75% of the total volume, and collecting the cut-off concentrated solution;
(3) purifying the ultrafiltration concentrated solution again by using ion exchange chromatography resin, concentrating and purifying the obtained intercepted concentrated solution by using macroporous adsorption resin at the flow rate of 4BV/h and 5 times of column accumulated water, wherein the elution flow rate is 1BV/h, then adjusting the pH value range to 7, and treating by using anion and cation exchange resin to connect a resin column in series; the mass ratio of the anion exchange resin to the cation resin is 1:5, so as to obtain a high-purity grifola frondosa polysaccharide extracting solution;
(4) spray drying the obtained high-purity grifola frondosa polysaccharide extract, and controlling the water content of finished product powder to be less than 20%, wherein the obtained dry powder is the grifola frondosa polysaccharide finished product.
Example 3:
(1) adding water 40 times the weight of the raw materials into the Grifola frondosa fruiting body raw material, extracting at 100 deg.C for 1 time, each time for 4 hr, filtering the extractive solution with 200 mesh sieve, and mixing the extractive solutions;
(2) ultrafiltering the Grifola frondosa extractive solution with molecular sieve cut-off membrane, with cut-off molecular weight of 20kD, working pressure of 1Mpa, temperature of 25 deg.C, stopping ultrafiltration when the volume of the permeate is 75% of the total volume, and collecting the cut-off concentrated solution;
(3) purifying the ultrafiltration concentrated solution again by using ion exchange chromatography resin, concentrating and purifying the obtained intercepted concentrated solution by using macroporous adsorption resin at the flow rate of 2BV/h and 5 times of column accumulated water, wherein the elution flow rate is 4BV/h, then adjusting the pH value range to 6.5, and treating by using anion and cation exchange resin to connect a resin column in series; the mass ratio of the anion exchange resin to the cation resin is 1:4, so that a high-purity grifola frondosa polysaccharide extracting solution is obtained;
(4) spray drying the obtained high-purity grifola frondosa polysaccharide extract, and controlling the water content of finished product powder to be less than 15%, wherein the obtained dry powder is the grifola frondosa polysaccharide finished product.
Comparing the content of the grifola frondosa polysaccharide extracted by the method with the content of the grifola frondosa polysaccharide obtained by extracting the grifola frondosa polysaccharide with high temperature water, the content of the grifola frondosa polysaccharide extracted by the method is 83.27 percent on average, and the content of the grifola frondosa polysaccharide extracted by extracting the grifola frondosa polysaccharide with high temperature water is only 26.12 percent on average.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (6)
1. A method for preparing and purifying a high-purity grifola frondosa polysaccharide extract is characterized by comprising the following steps:
(1) adding water 10-40 times the weight of Grifola frondosa fruiting body into the fruiting body, extracting at 80-105 deg.C for 1-4 times, each for 1-4 hr, filtering the extractive solution with 80-200 mesh sieve, and mixing the extractive solutions;
(2) ultrafiltering the Grifola frondosa extractive solution with molecular sieve cut-off membrane, with cut-off molecular weight of 10-80kD, working pressure of 0.3-1Mpa, temperature of 25-50 deg.C, stopping ultrafiltration when the volume of the permeate is 40-70% of the total volume, and collecting the cut-off concentrated solution;
(3) purifying the ultrafiltration concentrated solution again by using ion exchange chromatography resin, concentrating and purifying the obtained intercepted concentrated solution by using macroporous adsorption resin at the flow rate of 1-4BV/h and 3-10 times of cylinder accumulated water, wherein the elution flow rate is 0.5-5BV/h, then adjusting the pH value range to 6.5-8, and performing deproteinization treatment by using a cation and anion exchange resin series resin column; the mass ratio of the anion exchange resin to the cation resin is 1:1-5, so as to obtain a high-purity grifola frondosa polysaccharide extracting solution;
(4) drying the obtained high-purity grifola frondosa polysaccharide extract, controlling the moisture content of finished product powder to be less than 15%, and obtaining dry powder, namely the grifola frondosa polysaccharide finished product.
2. The extraction process as claimed in claim 1, wherein the drying of the ultrafiltration retentate comprises one or more of spray drying, freeze drying, vacuum drying or atmospheric drying.
3. The extraction method according to claim 1, wherein the microfiltration membrane material is one or more of a ceramic membrane, an organic polymer membrane and a metal membrane.
4. The extraction method according to claim 1, wherein the ultrafiltration membrane has a cut-off molecular weight of 10-80KDa, and the ultrafiltration membrane material is one or more of a ceramic membrane, an organic polymer membrane and a metal membrane.
5. The extraction method according to claim 1, wherein the ion exchange resin comprises one or more of anion exchange resin, cation exchange resin and nonionic exchange resin.
6. The use of the grifolan obtained by the extraction method according to claim 1 for enhancing immunity, resisting cancer, resisting virus, resisting inflammation, lowering blood sugar and lowering blood pressure.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110782672.2A CN113307893A (en) | 2021-07-12 | 2021-07-12 | Preparation and purification method of high-purity grifola frondosa polysaccharide extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110782672.2A CN113307893A (en) | 2021-07-12 | 2021-07-12 | Preparation and purification method of high-purity grifola frondosa polysaccharide extract |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113307893A true CN113307893A (en) | 2021-08-27 |
Family
ID=77381463
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110782672.2A Withdrawn CN113307893A (en) | 2021-07-12 | 2021-07-12 | Preparation and purification method of high-purity grifola frondosa polysaccharide extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113307893A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114149514A (en) * | 2022-01-05 | 2022-03-08 | 江苏华骏生物科技有限公司 | Extraction method and application of maitake mushroom extract |
-
2021
- 2021-07-12 CN CN202110782672.2A patent/CN113307893A/en not_active Withdrawn
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114149514A (en) * | 2022-01-05 | 2022-03-08 | 江苏华骏生物科技有限公司 | Extraction method and application of maitake mushroom extract |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101736053A (en) | Technique for extracting Grifola frondosa water-soluble polysaccharide | |
| CN106381319A (en) | High-efficiency extraction and separation method of grape seed proanthocyanidin oligomer | |
| CN104017098B (en) | A kind of method from mushroom waste mushroom stick extracting directly mushroom active polysaccharide | |
| CA1084488A (en) | Protein-bound polysaccharides | |
| WO2022077853A1 (en) | Method for extracting grifola frondosa active polysaccharide, and extracted active polysaccharide and use thereof | |
| WO2015103974A1 (en) | Method for extracting and purifying l-ergothioneine | |
| CN111228317A (en) | Preparation method and application of inonotus obliquus extract | |
| CN110916198A (en) | Method for simultaneously preparing pectic polysaccharide and viscous glycoprotein by using okra fermented wine lees | |
| CN106146652A (en) | A kind of method for extraction and purification of middle phycocyanin of delivering vegetables | |
| CN107722132B (en) | Method for co-producing algal polysaccharide and algal protein from algae | |
| CN113307893A (en) | Preparation and purification method of high-purity grifola frondosa polysaccharide extract | |
| CN104886595A (en) | Method for extracting flavonoids from peanut hulls with aqueous enzymatic method and alcohol and water solvent assisted by microwave | |
| CN106866787B (en) | Mushroom sobering-up peptide and preparation method and application thereof | |
| CA1072033A (en) | Method of producing nitrogen-containing polysaccharides | |
| CN104277129A (en) | Method for extracting grifola frondosa polysaccharides | |
| CN110882285A (en) | Efficient preparation method of active substances in phellinus igniarius | |
| CN108265092B (en) | Mushroom oligosaccharide with excellent antioxidant activity and preparation method thereof | |
| CN106467561A (en) | A kind of sharp separation, the method for purification Radix Ginseng arginine glycoside | |
| CN105367681A (en) | Edible and medicinal fungal polysaccharide low-temperature normal-pressure plasma extraction method | |
| CN1045472C (en) | Extracting polyse protein from | |
| CN114560959B (en) | Preparation method of mushroom extract | |
| CN104387486A (en) | High-yield preparation method of coriolus versicolor polysaccharide | |
| CN104262500A (en) | Novel Dictyophora rubrovolvata polysaccharide with immunocompetence, and preparation method and application thereof | |
| CN110194810A (en) | A kind of grading purification method for the external glue polysaccharide of water shield improving hypoglycemic activity | |
| CN107236054B (en) | Preparation method and application of low-molecular-weight ascophyllan |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20210827 |
|
| WW01 | Invention patent application withdrawn after publication |