CN103333268A - Method for preparing lycium ruthenicum polysaccharide - Google Patents
Method for preparing lycium ruthenicum polysaccharide Download PDFInfo
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- CN103333268A CN103333268A CN2013103084377A CN201310308437A CN103333268A CN 103333268 A CN103333268 A CN 103333268A CN 2013103084377 A CN2013103084377 A CN 2013103084377A CN 201310308437 A CN201310308437 A CN 201310308437A CN 103333268 A CN103333268 A CN 103333268A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
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- 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
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- 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
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- C—CHEMISTRY; METALLURGY
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- C—CHEMISTRY; METALLURGY
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
Abstract
The invention relates to a method for extracting and preparing lycium ruthenicum polysaccharide, which solves the problems of the existing method that the raw material cannot be sufficiently utilized, the cost is high, the yield of polysaccharide is low and the activity is poor. The method comprises the following steps of in the preparation process, adopting an ultra-fine crushing technique, crushing the lycium ruthenicum residues with pigment extracted, then degreasing to obtain the degreased lycium ruthenicum powder, enzymatically decomposing the degreased powder, adopting a freezing and thawing technique to repetitively freeze and thaw for 3 to 7 times, and finally carrying out filtering, alcohol deposition, deproteinization treatment, freeze-drying and the like, so as to obtain the lycium ruthenicum polysaccharide finished product with anti-oxidative activity. The method has the advantage that the lycium ruthenicum residues with pigment extracted are used as the raw material, and the ultra-fine crushing technique and the low-temperature freezing and thawing technique are adopted to prepare the lycium ruthenicum polysaccharide, so the yield is high, the activity is high, and the cost is low.
Description
Technical field:
The invention belongs to black fruit lyceum deep processing field, relevant with the extracting method of vegetable polysaccharides, especially be that the method for black fruit lyceum polysaccharide of feedstock production anti-oxidant activity is relevant to extract black fruit lyceum residue behind the pigment.
Technical background:
Black fruit lyceum is the mature fruit of Solanaceae (Solanaceae) Lycium (Lycium) plant black fruit lyceum.Black fruit lyceum covers " Qiao Nuoying-Ha Er mug(unit of measure) ", Tibetan medicine name " other agate " by name.The black fruit lyceum flavor is sweet, property flat, multiple nutritional components such as rich in proteins, lycium barbarum polysaccharide, amino acid, VITAMIN, mineral substance, trace element.Wherein contained lycium barbarum polysaccharide have regulate immunizing power, remove free radical, suppress tumor growth, delay senility, the effect of reducing blood-fat, hypoglycemic and antifatigue, have bigger value of exploiting and utilizing and market outlook.
Lycium barbarum polysaccharide is a kind of of vegetable polysaccharides, and the compound that vegetable polysaccharides is made up of with α one or β one glycosidic link many identical or different monose is prevalent in the natural plant body, comprises starch, Mierocrystalline cellulose, saccharan, pectin etc.Because the wide material sources of vegetable polysaccharides, the molecule of vegetable polysaccharides not of the same race constitutes and molecular weight has nothing in common with each other, and its corresponding biological activity and effect also are not quite similar.Lycium barbarum polysaccharide is mainly derived from fruits such as Lycium plant matrimony vine, black fruit lyceum.
The conventional extracting method of vegetable polysaccharides comprises that water extraction and alcohol precipitation method, microwave extraction method, soda acid decompose extraction method etc., comprise new technology and novel methods such as ultrasonic-assisted extraction, ultrafiltration process, supercritical extraction method in addition at present.Ordinary method need be passed through pyroprocessing mostly, be unfavorable for protecting active polysaccharide, and the new technology that developed recently gets up is higher to equipment requirements, and this has increased polysaccharide undoubtedly and has extracted the cost of preparation.
Chinese patent CN101029088 discloses a kind of preparation method of lycium barbarum polysaccharide, and it is raw material that this patent adopts medlar fresh fruit, utilizes the column chromatographic isolation and purification lycium barbarum polysaccharide.Whole technology comprises operations such as immersion, fragmentation, extraction, enzymolysis, column chromatography, alcohol precipitation, dealcoholysis, drying, utilize polygalacturonase to handle extracting solution, improved purity of polysaccharide, but selected enzyme is comparatively single, be broken for conventional broken, fail to improve polysaccharide yield and anti-oxidant activity, raw material is underused.
Summary of the invention:
The purpose of this invention is to provide that a kind of technology is simple, easy to operate, cost is low, polysaccharide extract rate is high, the polysaccharide anti-oxidant activity is strong, the preparation method of the sufficient black fruit lyceum polysaccharide of black fruit lyceum utilization.
The present invention is achieved by the following technical solutions:
The preparation method of black fruit lyceum polysaccharide, adopting the black fruit lyceum waste residue that has extracted pigment is raw material, its preparation process is as follows:
(1) the black fruit lyceum residue that will extract pigment is pulverized with the superfine comminution at low temperature machine dry below 55 ℃, and the particle after the pulverizing has 95% above particle by 300 order medicines sieve;
(2) the black fruit lyceum dried powder of getting after sieving places the extraction vessel, adds 10-15 times of amount petroleum ether extractions 1-3 times, and each 1-2 hours, cross the leaching filter residue, drying makes the sherwood oil volatilization clean, obtains defatted seed flour;
(3) cellulase, polygalacturonase being dissolved in pH is in 4-6 buffered soln, is mixed with mass percentage concentration and is 1.2%-3.0% cellulase, polygalacturonase mixing solutions, and described damping fluid is citric acid, Trisodium Citrate cocktail buffer;
(4) defatted seed flour in the step (2) is placed extractor, add 8-15 times of amount cellulases, polygalacturonase mixing solutions, extracting at constant temperature under 45-65 ℃ of conditions (the too high enzyme of temperature can lose activity) 10-50min gets mixture;
(5) mixture in the step (4) is placed cryogenic refrigerator, take out behind-80 ℃ of quick freezing 4-8min and at room temperature thaw, so freeze-thaw and circulate repeatedly 3-7 times;
(6) mixture of handling through multigelation in the filtration step (5) is got filtrate;
(7) filtrate in the evaporation step (6), temperature is controlled at 55-65 ℃, is concentrated into filtrate volume 8-12% and gets concentrated solution, slowly adds 3-7 times of amount dehydrated alcohols in concentrated solution, and constantly stirs, and then at 3-6 ℃, leaves standstill 8-16 hours, gets precipitation solution;
(8) precipitation solution in the centrifugation step (7), collecting precipitation, and with absolute ethanol washing 2-5 times, freeze-drying namely gets the black fruit lyceum polysaccharide crude then;
(9) employing polymeric amide chromatogram column technique or Sevag method are sloughed the protein in the black fruit lyceum Crude polysaccharides, are concentrated to mass percent 7-9% then, and vacuum lyophilization obtains the black fruit lyceum polysaccharide.
Micronizing can be processed into micron even nano level micro mist to starting material.Superfine communication technique can greatly the destruction cellularstructure, even break cell walls, make to be present in intracellular polysaccharide component and to come out, can improve polysaccharide extract rate.Simultaneously, micronizing also can change the molecular weight, glycosidic link ratio of polysaccharide molecule in the extract etc., and then changes water-soluble, the biological characteristics etc. of polyoses extract.
Vegetable polysaccharides is present in the plant cell wall mostly, cell walls mainly is made of materials such as Mierocrystalline cellulose, hemicellulose, pectin, select for use cellulase, polygalacturonase that Chinese medicinal materials is carried out pre-treatment, can decompose cell wall constituent, destroy cell wall structure, produce partial collapse, dissolving, loose, reduced when extracting the resistance from cell walls, intercellular substance, can accelerate the speed of effective constituent stripping cell, improve extraction efficiency.
Can be because the cell internal and external temperature changes the uneven huge temperature difference that produces in the low temperature frozen-thaw process, the thermal shock effect that this temperature difference is brought can effectively destroy cellularstructure, and then make the abundant stripping of intracellular polysaccharide component, can effectively improve the polysaccharide leaching yield, and low temperature freeze thawing leaching process carries out all the time at a lower temperature, has effectively reduced the loss of polysaccharide active components.
The present invention adopts superfine communication technique that material is carried out pre-treatment, greatly reduces the extract granularity, has increased itself and the contact area of extracting solvent, has improved the dissolution rate of polysaccharide.The present invention also is aided with the plurality of enzymes treatment technology, impels the decomposition of pectin, Mierocrystalline cellulose etc., makes the extraction of polysaccharide more thorough, and purity is higher.More crucial is that the present invention finally uses the method for low temperature freeze thawing to destroy cellularstructure, accelerates the stripping of polysaccharide component in the cell, effectively improves active polysaccharide and yield.
Advantage of the present invention is as follows:
1, the present invention is raw material with the black fruit lyceum residue that extracted pigment, has taken full advantage of black fruit lyceum, makes its value maximization, has avoided unnecessary waste.For the comprehensive development and utilization black fruit lyceum provides feasible method.
2, the present invention adopts the super-fine powder technology, has improved extraction efficiency, has reduced the molecular weight that extracts polysaccharide, makes the polysaccharide anti-oxidant activity of preparation higher.Compare with common pulverizing extraction, under identical extraction conditions, extract after the micronizing, the yield of polysaccharide increases about 27%.The polysaccharide that the polysaccharide that extracts after the micronizing is compared common pulverizing extraction is high by 18.3% to the clearance rate of DPPH free radical
3, cellulase, polygalacturonase are dissolved in pH is in 4-6 the buffered soln in the present invention, be mixed with concentration and be 1.5% cellulase, polygalacturonase mixing solutions, cellulase and polygalacturonase react all needs the PH that suits, optimum pH is between 4-6, use this damping fluid that reliable and stable PH environment can be provided, be conducive to enzyme performance maximum effect, impel the decomposition of pectin, Mierocrystalline cellulose etc., make the extraction of polysaccharide more thorough, purity is higher.
4, the present invention has also adopted the low temperature freeze-thaw technology, utilizes to freeze-thermal shock effect that produces in the course of defrosting destroys cellularstructure, and then reaches and increase polysaccharide yield and active purpose.
Embodiment:
Embodiment 1:
1. will extract the black fruit lyceum residue drying (below 55 ℃) of pigment, and pulverize with the superfine comminution at low temperature machine, the particle after the pulverizing has 95% above particle by 300 order medicines sieve;
2. the black fruit lyceum dried powder of getting after sieving places the extraction vessel, adds 10 times of amount petroleum ether extractions 3 times, and each 1 hour, cross the leaching filter residue, drying makes the sherwood oil volatilization clean, obtains defatted seed flour;
3. cellulase, polygalacturonase being dissolved in pH is in 4 buffered soln, is mixed with mass percentage concentration and is 2.0% cellulase, polygalacturonase mixing solutions, and described damping fluid is citric acid, Trisodium Citrate cocktail buffer;
4. the defatted seed flour in the step (2) is placed extractor, add 8 times of amount cellulases, polygalacturonase mixing solutions, extracting at constant temperature 10min under 45 ℃ of conditions gets mixture;
5. the mixture in the step (4) is placed to take out behind cryogenic refrigerator (-80 take the photograph ℃) the quick freezing 6min under 25 ℃ of room temperatures and thaw, so freeze-thaw and circulate 3 times repeatedly;
6. the mixture of handling through multigelation in the filtration step (5) is got filtrate;
7. the filtrate in the rotary evaporation step (6), temperature is controlled at 55 ℃, be concentrated into filtrate volume 8% concentrated solution, in concentrated solution, slowly add 3 times of amount dehydrated alcohols, and constantly stir, then at 5 ℃, leave standstill 8 hours alcohol precipitations;
8. the precipitation solution in the centrifugation step (7), collecting precipitation, and with absolute ethanol washing 2 times, freeze-drying namely gets the black fruit lyceum polysaccharide crude then;
9. employing polymeric amide chromatogram column technique or Sevag method are sloughed the protein in the black fruit lyceum Crude polysaccharides, and being concentrated to mass percent then is 7%, and vacuum lyophilization obtains the black fruit lyceum polysaccharide.
Embodiment 2:
1. will extract the black fruit lyceum residue drying (below 55 ℃) of pigment, and pulverize with the superfine comminution at low temperature machine, the particle after the pulverizing has 95% above particle by 300 order medicines sieve;
2. the black fruit lyceum dried powder of getting after sieving places the extraction vessel, adds 15 times of amount petroleum ether extractions 3 times, and each 2 hours, cross the leaching filter residue, drying makes the sherwood oil volatilization clean, obtains defatted seed flour;
3. cellulase, polygalacturonase being dissolved in pH is in 6 buffered soln, is mixed with mass percentage concentration and is 1.5% cellulase, polygalacturonase mixing solutions, and described damping fluid is citric acid, Trisodium Citrate cocktail buffer;
4. the defatted seed flour in the step (2) is placed extractor, add 15 times of amount cellulases, polygalacturonase mixing solutions, extracting at constant temperature 50min under 65 ℃ of conditions gets mixture;
5. the mixture in the step (4) is placed to take out behind cryogenic refrigerator (-80 take the photograph ℃) the quick freezing 6min under 27 ℃ of room temperatures and thaw, so freeze-thaw and circulate 7 times repeatedly;
6. the mixture of handling through multigelation in the filtration step (5) is got filtrate;
7. the filtrate in the rotary evaporation step (6), temperature is controlled at 65 ℃, be concentrated into filtrate volume 12% concentrated solution, in concentrated solution, slowly add 7 times of amount dehydrated alcohols, and constantly stir, then at 4 ℃, leave standstill 16 hours alcohol precipitations;
8. the precipitation solution in the centrifugation step (7), collecting precipitation, and with absolute ethanol washing 5 times, freeze-drying namely gets the black fruit lyceum polysaccharide crude then;
9. employing polymeric amide chromatogram column technique or Sevag method are sloughed the protein in the black fruit lyceum Crude polysaccharides, and being concentrated to mass percent then is 9%, and vacuum lyophilization obtains the black fruit lyceum polysaccharide.
Embodiment 3:
1. will extract the black fruit lyceum residue drying (below 55 ℃) of pigment, and pulverize with the superfine comminution at low temperature machine, the particle after the pulverizing has 95% above particle by 300 order medicines sieve;
2. the black fruit lyceum dried powder of getting after sieving places the extraction vessel, adds 12 times of amount petroleum ether extractions 2 times, and each 1.5 hours, cross the leaching filter residue, drying makes the sherwood oil volatilization clean, obtains defatted seed flour;
3. cellulase, polygalacturonase being dissolved in pH is in 5.5 buffered soln, is mixed with mass percentage concentration and is 2.8% cellulase, polygalacturonase mixing solutions, and described damping fluid is citric acid, Trisodium Citrate cocktail buffer;
4. the defatted seed flour in the step (2) is placed extractor, add 10 times of amount cellulases, polygalacturonase mixing solutions, extracting at constant temperature 40min under 55 ℃ of conditions;
5. the mixture in the step (4) is placed to take out behind cryogenic refrigerator (80 take the photograph ℃) the quick freezing 6min under 27 ℃ of room temperatures and thaw, so freeze-thaw and circulate 4 times repeatedly;
6. the material of handling through multigelation in the filtration step (5) is got filtrate;
7. the filtrate in the rotary evaporation step (6), temperature is controlled at 60 ℃, be concentrated into filtrate volume 10% concentrated solution, in concentrated solution, slowly add 4 times of amount dehydrated alcohols, and constantly stir, then at 3 ℃, leave standstill 12 hours alcohol precipitations;
8. the precipitation solution in the centrifugation step (7), collecting precipitation, and with absolute ethanol washing 4 times, freeze-drying namely gets the black fruit lyceum polysaccharide crude then;
9. employing polymeric amide chromatogram column technique or Sevag method are sloughed the protein in the black fruit lyceum Crude polysaccharides, and being concentrated to mass percent then is 8%, and vacuum lyophilization obtains the black fruit lyceum polysaccharide.
Experimental example 1 Different Extraction Method is to the influence of black fruit lyceum polysaccharide extract rate
Get the 10g black fruit lyceum respectively and adopt water extraction and alcohol precipitation method, microwave extraction method, enzyme extraction method, micronizing method and the inventive method to extract respectively, measure polysaccharide yield separately then, as following table:
Table 1 Different Extraction Method is extracted the yield of black fruit lyceum polysaccharide
| Extracting method | Water extract-alcohol precipitation | Microwave extraction | Two enzyme extraction | Micronizing is extracted | The present invention |
| The polysaccharide yield | 2.89% | 3.05% | 3.15% | 3.66% | 4.12% |
The black fruit lyceum lycium barbarum polysaccharide that experimental example 2 Different Extraction Method are extracted is to the removing effect of DPPH
The black fruit lyceum lycium barbarum polysaccharide that table 2 Different Extraction Method is extracted is to the clearance rate of DPPH
| Extracting method | Water extract-alcohol precipitation | Microwave extraction | Two enzyme extraction | Micronizing is extracted | The present invention |
| Clearance rate | 37.5% | 41.3% | 45.6% | 51.2% | 72.1% |
Table 3 citric acid-sodium citrate buffer solution (0.1mol/L) configuration proportion
Remarks: citric acid C 6 H8 O7H 2 O, relative molecular mass=210.14; 0.1 mol/L solution is 21.01 g/L.
Trisodium Citrate Na3C 6H 5 O72H 2O, relative molecular mass=294.12; 0.1 mol/L solution is 29.41 g/L.
Claims (1)
1. the preparation method of black fruit lyceum polysaccharide, it is characterized in that adopting the black fruit lyceum waste residue that has extracted pigment is raw material, its preparation process is as follows:
(1) the black fruit lyceum residue that will extract pigment is pulverized with the superfine comminution at low temperature machine dry below 55 ℃, and the particle after the pulverizing has 95% above particle by 300 order medicines sieve,
(2) the black fruit lyceum dried powder of getting after sieving places the extraction vessel, adds 10-15 times of amount petroleum ether extractions 1-3 times, and each 1-2 hours, cross the leaching filter residue, drying makes the sherwood oil volatilization clean, obtains defatted seed flour,
(3) cellulase, polygalacturonase being dissolved in pH is in 4-6 buffered soln, is mixed with mass percentage concentration and is 1.2%-3.0% cellulase, polygalacturonase mixing solutions, and described damping fluid is citric acid, Trisodium Citrate cocktail buffer,
(4) defatted seed flour in the step (2) is placed extractor, add 8-15 times of amount cellulases, polygalacturonase mixing solutions, extracting at constant temperature 10-50min gets mixture under 45-65 ℃ of conditions,
(5) mixture in the step (4) is placed cryogenic refrigerator, takes out behind-80 ℃ of quick freezing 4-8min and at room temperature thaw, so freeze-thaw and circulate repeatedly 3-7 times,
(6) mixture of handling through multigelation in the filtration step (5) is got filtrate,
(7) filtrate in the rotary evaporation step (6), temperature is controlled at 55-65 ℃, is concentrated into filtrate volume 8-12% and gets concentrated solution, in concentrated solution, slowly add 3-7 times of amount dehydrated alcohols, and constantly stir, then at 3-6 ℃, left standstill 8-16 hours, and got precipitation solution
(8) precipitation solution in the centrifugation step (7), collecting precipitation, and with absolute ethanol washing 2-5 times, freeze-drying namely gets the black fruit lyceum polysaccharide crude then,
(9) employing polymeric amide chromatogram column technique or Sevag method are sloughed the protein in the black fruit lyceum Crude polysaccharides, are concentrated to mass percent 7-9% then, and vacuum lyophilization obtains the black fruit lyceum polysaccharide.
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| CN201310308437.7A CN103333268B (en) | 2013-07-23 | 2013-07-23 | The preparation method of lycium ruthenicum polysaccharide |
| PCT/CN2014/077363 WO2015010497A1 (en) | 2013-07-23 | 2014-05-13 | Method for preparing lycium ruthenicum polysaccharide |
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| WO2015010497A1 (en) * | 2013-07-23 | 2015-01-29 | 四川圣湖生物科技有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN105211867A (en) * | 2015-10-28 | 2016-01-06 | 青海尤尼克生物技术有限公司 | A kind of black fruit lyceum compound fruit ferment and preparation method thereof |
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| WO2015010497A1 (en) * | 2013-07-23 | 2015-01-29 | 四川圣湖生物科技有限公司 | Method for preparing lycium ruthenicum polysaccharide |
| CN104222889A (en) * | 2014-09-10 | 2014-12-24 | 中国科学院过程工程研究所 | Wolfberry soluble dietary fiber and preparation method thereof |
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| CN109503729A (en) * | 2018-12-12 | 2019-03-22 | 湖南医药学院 | A method of extracting polysaccharide |
| CN110305231A (en) * | 2019-06-24 | 2019-10-08 | 温州广立生物医药科技有限公司 | A kind of the extraction preparation method and its extract equipment of lycium ruthenicum polysaccharide |
| CN110256599A (en) * | 2019-07-18 | 2019-09-20 | 成都师范学院 | A kind of enzyme assisted extraction lycium ruthenicum polysaccharide process |
| CN111925457A (en) * | 2020-07-24 | 2020-11-13 | 哈工大机器人南昌智能制造研究院 | Method for preparing high-purity lycium ruthenicum polysaccharide |
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| WO2015010497A1 (en) | 2015-01-29 |
| CN103333268B (en) | 2016-01-27 |
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