CN103059156A - Method for extracting tea polysaccharide from tea seed cake - Google Patents
Method for extracting tea polysaccharide from tea seed cake Download PDFInfo
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Abstract
The invention discloses a method for extracting tea polysaccharide from tea seed cake. The invention relates to the technical field of agricultural product deep processing. The extraction process comprises the steps such as pretreatment, mixed enzyme preparation, enzyme processing, pressing, primary drying, eluting, precipitating, alkali dissolving, centrifugation, column I processing, column II processing, sedimentation, dissolving, secondary drying, and the like. With the method, tea polysaccharide can be extracted from tea seed cake, and extraction purity is high. With the method, tea polysaccharide can be extracted from tea seed cake, and can be extracted from tea seeds or tea leaves. Also, tea polysaccharide can be extracted from other plants comprising tea polysaccharide.
Description
Technical field
The present invention relates to deep-processing technical field of agricultural products, particularly a kind of method by the withered middle extraction tea polysaccharide of tea.
Technical background
Tea polysaccharide (Tea Polysaccharide) is a kind of macromolecular compound (molecular weight 40,000-100,000) of a kind of similar ganoderan and panaxan, it is a kind of acid glycoprotein, and be combined with a large amount of mineral elements, be called the Tea Polysaccharides mixture, referred to as Tea Polysaccharides or tea polysaccharide.Its protein part mainly is comprised of about 20 kinds of common amino acid, and the part of sugar is mainly by pectinose, wood sugar, Fucose, glucose, semi-lactosi etc., and mineral element is mainly by calcium, magnesium, iron, manganese etc. and a small amount of trace element, such as compositions such as rare earth elements.Have the effects such as hypoglycemic, reducing blood-fat, strengthening immunity, hypotensive, reducing heart rate, increase coronary flow, anticoagulation, antithrombotic and hypoxia tolerance, the discovered in recent years tea polysaccharide also has the effect for the treatment of diabetes.Launch the research of polysaccharide is begun from 20 century 70s, develop very fast.Because the diversified bioactive functions of polysaccharide and at functional food be widely used clinically becomes increasingly active the explo iting and researching of polysaccharide Biological resources, becomes the study hotspot of natural drug, biological chemistry, life science.Process in advance: the tea polysaccharide main source is the dried tea of finished product.By tealeaves is pulverized, at a certain temperature, use first acidity, neutrality or alkalescent water lixiviate, by precipitate and separate or column chromatography for separation, prepare crude product again.At present, the preparation of tea polysaccharide mainly is by extracting in tealeaves or the tea seed oil cake, and method mainly contains following three kinds:
The one, organic solvent precipitation method: the solubleness of tea polysaccharide in organic solvent is less, can be with tea polysaccharide and other composition initial gross separations by the method for organic solvent deposit; The organic solvent of using mainly contains ethanol, acetone, ethyl acetate etc.
The 2nd, quaternary salt deposit method: under acidity, neutrality or weak basic condition, some quaternary ammonium salts such as Cetyltrimethyl Ammonium Bromide (CTAB), hexadecyl pyridine hydrochloride (CPC) can form water-fast salt with tea polysaccharide, with this tea polysaccharide and other component separating, the recycling dialysis method obtains than sterling.
The 3rd, resin method: tea polysaccharide is a kind of acidic polysaccharose, and acidic polysaccharose generally can both be adsorbed by associated resin, such as the DEAE-fibre resin.Then tea polysaccharide and other compositions can be separated by different elutriants.
Chinese patent (number of patent application is 201010259131.3) " combined extraction method of tea saponin, tea polysaccharide in camellia seed meal or the tea-seed dregs of rice ", take degreasing tea seed grouts, tea-seed grouts as raw material, the integrated extraction technique of research tea saponin, tea polysaccharide; Its extracting method is: camellia seed meal or the tea-seed dregs of rice are pulverized and are obtained thick tea saponin and tea seed polysaccharide by water extract-alcohol precipitation or alcohol extracting water extraction, clear liquid after the water extraction or after the alcohol extracting is through the macroporous adsorption resin chromatography column purification, the resin absorption thing is first with alkali, the ethanol elution that increases gradually with volumetric concentration again, collect ethanol eluate, can obtain highly purified tea saponin; Present method technique is simple, and production cost is low, not only can prepare highly purified tea saponin, also can obtain simultaneously another active result tea polysaccharide.And for example Chinese patent (number of patent application is 200710191396.2) " extracts the method for tea polysaccharide " from camellia seed meal, in the ratio of 10kg than 100-200 premium on currency with camellia seed meal, in water, soak, then heat and stir simultaneously, filter while hot, liquid after filtering is continued heating, make liquid concentration to 1/4 volume, the yeast that adds in concentrated solution remains under the 25-32 ℃ of temperature, the vibration fermentation, after fermentation time 0.5-3 days, stop vibration, make its precipitation, the ethanol of getting 3 times of supernatant liquid adding clear liquid volumes stirs, then put into refrigerator freezing, refrigerating fulid is added water make its dissolving layering and precipitating, get supernatant liquid, with the spray-dried tea polysaccharide that gets of supernatant liquid; This invention has solved the method for extracting at present tea polysaccharide from camellia seed meal, complex process, and production unit is many, the deficiency of cost height.
Technology contents
The object of the present invention is to provide a kind of can be from tea be withered the method for extraction tea polysaccharide, have the higher characteristics of dna purity.
For realizing its purpose, the technical solution used in the present invention is a kind of method by the withered middle extraction tea polysaccharide of tea of invention, and its leaching process is as follows:
A, pre-treatment: select witheredly without mould bad tea, being ground into granularity is the withered particle of tea of 10-35mm, then, drops in the fermenting container, adds the clear water that accounts for the withered particle weight of tea 30-50% again, stirs evenly, and becomes the withered slurry of tea, for subsequent use;
B, mixed enzyme preparation:
Raw material with following weight part:
Saccharifying enzyme 5-10 hydrolysising protease 4-8 flour 5-10
Cellulase 3-5 wheat-flour 10-15 polyphenoloxidase 3-5
Mix, become mixed enzyme, for subsequent use;
Described saccharifying enzyme, hydrolysising protease, cellulase and polyphenoloxidase are respectively α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme, BA13ZXDBM hydrolysising protease, GFDS-23 cellulase and E0145 polyphenoloxidase;
C, enzyme are processed: in the withered slurry of tea for subsequent use, add the mixed enzyme that accounts for the withered particle weight of tea 6-10%, heat to 20-40 ℃, stir evenly, and encapsulation process 4-6 days, become enzyme and process the withered slurry of tea, for subsequent use;
D, squeezing: process in the withered slurry of tea to enzyme for subsequent use, add the 40-65 ℃ of warm water that accounts for the withered particle weight of tea 10-30%, stir evenly, at normal temperatures, squeeze processing, collect the liquid of squeezing out, become the enzyme treatment solution, for subsequent use;
E, primary drying: enzyme treatment solution for subsequent use is heated to 70-100 ℃, and insulation was processed 6-10 hour, added edible ethanol again, made the volumetric concentration of enzyme treatment solution alcohol reach 50%, then, 4-12 ℃ of refrigeration 10-15 hour, filtered, and collected mixing liquid; Again with mixing liquid at 40-90 ℃ of vacuum concentration, concentration time is 2-3.5 hour, gets concentrated solution; Spray-dried again, get dry powder, for subsequent use;
F, wash-out: add in the dry powder for subsequent use account for dry powder weight 1-2 doubly, volumetric concentration is 95% polar organic solvent, leaves standstill after stirring evenly 2-4 hour, filters, and isolates polar organic solvent, so process twice; Then, again to recrement add account for recrement weight 1-2 doubly, volumetric concentration is the low polar organic solvent that (perhaps is neat solvent) 95% or more, leaves standstill after stirring evenly 1-2 hour, filtration is so processed three times, the collection and treatment slag is for subsequent use;
Described polar organic solvent is a kind of in ethanol, methyl alcohol, ethyl acetate, the propyl carbinol or two kinds or multiple, and when being two or more, the proportioning of each composition is single-candidate or other ratio;
Described low polar organic solvent is normal hexane or sherwood oil or both mixtures, and when being two kinds, both proportionings are single-candidate or other ratio;
G, precipitation: add in the processing slag for subsequent use and account for that to process slag weight 20-40%, volumetric concentration be 95% methyl alcohol, add again to account for the saturated lead sulfate aqueous solution of processing slag weight 5-12%, processed 8-12 hour; And then adding accounts for processing slag weight 3-5%, concentration is the ferric sulfate aqueous solution of 10-25%, processes 0.8-1.2 hour, filters, and isolates liquid, and collecting precipitation is for subsequent use;
H, alkali are molten: in precipitation for subsequent use, add precipitation weight 0.5-1.5 doubly, concentration is the weak caustic solution of 1.5-2.4%, dissolving 20-40min; Then, add again the 60-85 ℃ of warm water that 3-5 doubly precipitates weight, soak and extract, filter, collect the aqueous solution, for subsequent use;
Described weak caustic solution is sodium soap or sodium bicarbonate or sodium carbonate solution;
I, centrifugal: with the aqueous solution for subsequent use through 5000-10000 turn/supercentrifuge of min is centrifugal, gets centrifugate, and is for subsequent use;
J, mistake post I: through the mixing column filtration treatment, coutroi velocity is 50-70L/h with centrifugate for subsequent use, collects filtrate, and is for subsequent use;
K, mistake post II: filtrate for subsequent use is crossed post through polar macroporous resin process, coutroi velocity is 15-25L/h, collects post liquid, and is for subsequent use;
The model of described polar macroporous resin is D301;
L, sediment: for subsequent use crossed post liquid at 70-100 ℃ vacuum concentration to 1/3, became the post concentrated solution, then add gradually the organic solvent miscible with water, till when the concentration of the miscible organic solvent of transference water reaches 70-95%, mix thoroughly, add again volume and be post concentrated solution 1-2 doubly, with water-insoluble organic solvent, stir process 20-36 hour, leave standstill, layering, collect lower floor's insolubles, for subsequent use;
The miscible organic solvent of described and water is ethanol or methyl alcohol;
Described and water-insoluble organic solvent is ethyl acetate or normal hexane or sherwood oil;
M, dissolving: insolubles for subsequent use with 1-3 normal-temperature water dissolving doubly, is filtered, collect lysate, for subsequent use;
N, redrying: with lysate for subsequent use, spraying drying gets the tea polysaccharide finished product;
Mixing column in the J step is two kinds or multiple mixture in neutral gac or quartz sand or diatomite or the polymeric adsorbent.
The technical scheme one of its optimization: described mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 5 hydrolysising proteases 8 flour 5
Cellulase 3 wheat-flours 15 polyphenoloxidase 5.
The technical scheme two of its optimization: described mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 10 hydrolysising proteases 4 flour 10
Cellulase 5 wheat-flours 10 polyphenoloxidase 3.
The technical scheme three of its optimization: described mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 8 hydrolysising proteases 6 flour 8
Cellulase 4 wheat-flours 12 polyphenoloxidase 4.
Method by the withered middle extraction tea polysaccharide of tea of the present invention, to adopt the tea squeezed behind the camellia seed oil withered as raw material, adopt pre-treatment, mixed enzyme preparation, enzyme processing, squeezing, primary drying, wash-out, precipitation, alkali molten, centrifugal, cross post I, steps such as post II, sediment, dissolving, redrying excessively, contained tea polysaccharide during tea is withered has been proposed effectively, and removed impurity in the tea polysaccharide finished product, thereby the tea polysaccharide content in its finished product reaches more than 95%, and purity is higher.Simultaneously, by the extraction of tea polysaccharide, enlarge the withered Application Areas of tea, improved the value of the withered utilization of tea.
Below table 1 be the tea polysaccharide extracted according to the inventive method and the detection data contrast of the major ingredient content of the tea polysaccharide that adopts ordinary method to extract.
The detection synopsis of the major ingredient content of table 1 tea polysaccharide
Data can be found out in the top table, and according to the tea polysaccharide that the inventive method is extracted, the content of its various effective ingredients far surpasses the tea polysaccharide that ordinary method is extracted.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated.Following explanation is the mode that exemplifies, but protection scope of the present invention is not limited to this.
Method by the withered middle extraction tea polysaccharide of tea of the present invention, used mixed enzyme are to be mixed by the raw material with following weight part:
Saccharifying enzyme 5-10 hydrolysising protease 4-8 flour 5-10
Cellulase 3-5 wheat-flour 10-15 polyphenoloxidase 3-5.
Embodiment one
Leaching process is as follows:
A, pre-treatment: select witheredly without mould bad tea, being ground into granularity is the withered particle of tea of 10-35mm, then, drops in the fermenting container, adds the clear water that accounts for the withered particle weight 40% of tea again, stirs evenly, and becomes the withered slurry of tea, for subsequent use;
B, mixed enzyme preparation:
Raw material with following weight part:
Saccharifying enzyme 5 hydrolysising proteases 8 flour 5
Cellulase 3 wheat-flours 15 polyphenoloxidase 5
Mix, become mixed enzyme, for subsequent use;
Used saccharifying enzyme, hydrolysising protease, cellulase and polyphenoloxidase are respectively α-Isosorbide-5-Nitrae-glucose hydrolysis enzyme, BA13ZXDBM hydrolysising protease, GFDS-23 cellulase and E0145 polyphenoloxidase;
C, enzyme are processed: in the withered slurry of tea for subsequent use, add the mixed enzyme that accounts for the withered particle weight 8% of tea, heat to 30 ℃, stir evenly, encapsulation process 5 days becomes enzyme and processes the withered slurry of tea, and is for subsequent use;
D, squeezing: process in the withered slurry of tea to enzyme for subsequent use, add 50 ℃ of warm water that account for the withered particle weight 20% of tea, stir evenly, at normal temperatures, squeeze processing, collect the liquid of squeezing out, become the enzyme treatment solution, for subsequent use;
E, primary drying: enzyme treatment solution for subsequent use is heated to 85 ℃, and insulation was processed 8 hours, added edible ethanol again, made the volumetric concentration of enzyme treatment solution alcohol reach 50%, then, 6 ℃ of refrigerations 12 hours, filtered, and collected mixing liquid; Again with mixing liquid at 60 ℃ of vacuum concentration, concentration time is 3 hours, gets concentrated solution (be concentrated into 1/6 or density be 1.2); Spray-dried again, get dry powder, for subsequent use;
F, wash-out: add in the dry powder for subsequent use account for dry powder weight 1-2 doubly, volumetric concentration is 95% polar organic solvent, leaves standstill after stirring evenly 2-4 hour, filters, and isolates polar organic solvent, so process twice; Then, again to recrement add account for recrement weight 1-2 doubly, volumetric concentration is the low polar organic solvent that (perhaps is neat solvent) 95% or more, leaves standstill after stirring evenly 1-2 hour, filtration is so processed three times, the collection and treatment slag is for subsequent use;
Add that to account for 1.5 times of dry powder weight, volumetric concentration be 95% ethanol in the dry powder for subsequent use, left standstill after stirring evenly 3 hours, filter, isolate ethanol, so process twice; Then, add to recrement again that to account for 1.5 times of recrement weight, volumetric concentration be 95% normal hexane, left standstill after stirring evenly 1-2 hour, filter, so process three times, the collection and treatment slag, for subsequent use;
G, precipitation: add in the processing slag for subsequent use and account for that to process slag weight 30%, volumetric concentration be 95% methyl alcohol, add again to account for the saturated lead sulfate aqueous solution of processing slag weight 8%, processed 10 hours; And then add and to account for that to process slag weight 4%, concentration be 20% ferric sulfate aqueous solution, to process 1 hour, liquid is isolated in filtration, and collecting precipitation is for subsequent use;
H, alkali are molten: in precipitation for subsequent use, add 1 times of weight of precipitation, concentration and be 2.0% sodium hydrogen carbonate solution, dissolving 30min; Then, add again 70 ℃ of warm water of 4 times of precipitation weight, soak and extract, filter, collect the aqueous solution, for subsequent use;
I, centrifugal: with the aqueous solution for subsequent use through 5000-10000 turn/supercentrifuge of min is centrifugal, gets centrifugate, and is for subsequent use;
J, mistake post I: through mixing column (adopting the mixture of neutral gac and quartz sand) filtration treatment, coutroi velocity is 60L/h with centrifugate for subsequent use, collects filtrate, and is for subsequent use; Used post adopts stainless steel column, long 8m, aperture 0.8m;
K, mistake post II: filtrate for subsequent use is crossed post through polar macroporous resin (model is D301) process, coutroi velocity is 20L/h, collects post liquid, and is for subsequent use;
L, sediment: for subsequent use crossed post liquid at 85 ℃ of vacuum concentration to 1/3, became the post concentrated solution, then add gradually ethanol, till when allowing the concentration of ethanol reach 80%, mix thoroughly, adding volume was the ethyl acetate of 1.5 times of post concentrated solutions again, stir process 28 hours, leave standstill, layering, collect lower floor's insolubles, for subsequent use;
M, dissolving: with insolubles for subsequent use normal-temperature water dissolving with 2 times, filter, collect lysate, for subsequent use;
N, redrying: with lysate for subsequent use, spraying drying gets the tea polysaccharide finished product, and tea polysaccharide content wherein reaches 85.1%.
Embodiment two
The present embodiment and embodiment one are basic identical, and its difference is:
Mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 10 hydrolysising proteases 4 flour 10
Cellulase 5 wheat-flours 10 polyphenoloxidase 3;
Used mixing column is the mixture of neutral gac and quartz sand.
The tea polysaccharide finished product that extracts, its content are 85.2%.
Embodiment three
The present embodiment and embodiment one are basic identical, and its difference is:
Mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 8 hydrolysising proteases 6 flour 8
Cellulase 4 wheat-flours 12 polyphenoloxidase 4;
Used mixing column is the mixture of diatomite and polymeric adsorbent.
The tea polysaccharide finished product that extracts, its content are 85.3%.
Method by the withered middle extraction tea polysaccharide of tea of the present invention both can be used for extraction tea polysaccharide from tea is withered, also can be used for extracting tea polysaccharide from tea-seed or tealeaves, also can be used for extracting tea polysaccharide from other contains the plant of tea polysaccharide.
Claims (4)
1. method by the withered middle extraction tea polysaccharide of tea is characterized in that leaching process is as follows:
A, pre-treatment: select witheredly without mould bad tea, being ground into granularity is the withered particle of tea of 10-35mm, then, drops in the fermenting container, adds the clear water that accounts for the withered particle weight of tea 30-50% again, stirs evenly, and becomes the withered slurry of tea, for subsequent use;
B, mixed enzyme preparation:
Raw material with following weight part:
Saccharifying enzyme 5-10 hydrolysising protease 4-8 flour 5-10
Cellulase 3-5 wheat-flour 10-15 polyphenoloxidase 3-5
Mix, become mixed enzyme, for subsequent use;
C, enzyme are processed: in the withered slurry of tea for subsequent use, add the mixed enzyme that accounts for the withered particle weight of tea 6-10%, heat to 20-40 ℃, stir evenly, and encapsulation process 4-6 days, become enzyme and process the withered slurry of tea, for subsequent use;
D, squeezing: process in the withered slurry of tea to enzyme for subsequent use, add the 40-65 ℃ of warm water that accounts for the withered particle weight of tea 10-30%, stir evenly, at normal temperatures, squeeze processing, collect the liquid of squeezing out, become the enzyme treatment solution, for subsequent use;
E, primary drying: enzyme treatment solution for subsequent use is heated to 70-100 ℃, and insulation was processed 6-10 hour, added edible ethanol again, made the volumetric concentration of enzyme treatment solution alcohol reach 50%, then, 4-12 ℃ of refrigeration 10-15 hour, filtered, and collected mixing liquid; Again with mixing liquid at 40-90 ℃ of vacuum concentration, concentration time is 2-3.5 hour, gets concentrated solution; Spray-dried again, get dry powder, for subsequent use;
F, wash-out: add in the dry powder for subsequent use account for dry powder weight 1-2 doubly, volumetric concentration is 95% polar organic solvent, leaves standstill after stirring evenly 2-4 hour, filters, and isolates polar organic solvent, so process twice; Then, again to recrement add account for recrement weight 1-2 doubly, volumetric concentration is the low polar organic solvent that (perhaps is neat solvent) 95% or more, leaves standstill after stirring evenly 1-2 hour, filtration is so processed three times, the collection and treatment slag is for subsequent use;
G, precipitation: add in the processing slag for subsequent use and account for that to process slag weight 20-40%, volumetric concentration be 95% methyl alcohol, add again to account for the saturated lead sulfate aqueous solution of processing slag weight 5-12%, processed 8-12 hour; And then adding accounts for processing slag weight 3-5%, concentration is the ferric sulfate aqueous solution of 10-25%, processes 0.8-1.2 hour, filters, and isolates liquid, and collecting precipitation is for subsequent use;
H, alkali are molten: in precipitation for subsequent use, add precipitation weight 0.5-1.5 doubly, concentration is the weak caustic solution of 1.5-2.4%, dissolving 20-40min; Then, add again the 60-85 ℃ of warm water that 3-5 doubly precipitates weight, soak and extract, filter, collect the aqueous solution, for subsequent use;
I, centrifugal: with the aqueous solution for subsequent use through 5000-10000 turn/supercentrifuge of min is centrifugal, gets centrifugate, and is for subsequent use;
J, mistake post I: through the mixing column filtration treatment, coutroi velocity is 50-70L/h with centrifugate for subsequent use, collects filtrate, and is for subsequent use;
K, mistake post II: filtrate for subsequent use is crossed post through polar macroporous resin process, coutroi velocity is 15-25L/h, collects post liquid, and is for subsequent use;
L, sediment: for subsequent use crossed post liquid at 70-100 ℃ vacuum concentration to 1/3, became the post concentrated solution, then add gradually the organic solvent miscible with water, till when the concentration of the miscible organic solvent of transference water reaches 70-95%, mix thoroughly, add again volume and be post concentrated solution 1-2 doubly, with water-insoluble organic solvent, stir process 20-36 hour, leave standstill, layering, collect lower floor's insolubles, for subsequent use;
M, dissolving: insolubles for subsequent use with 1-3 normal-temperature water dissolving doubly, is filtered, collect lysate, for subsequent use;
N, redrying: with lysate for subsequent use, spraying drying gets the tea polysaccharide finished product;
Mixing column in the J step is two kinds or multiple mixture in neutral gac or quartz sand or diatomite or the polymeric adsorbent.
2. described method by the withered middle extraction tea polysaccharide of tea according to claim 1 is characterized in that described mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 5 hydrolysising proteases 8 flour 5
Cellulase 3 wheat-flours 15 polyphenoloxidase 5.
3. described method by the withered middle extraction tea polysaccharide of tea according to claim 1 is characterized in that described mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 10 hydrolysising proteases 4 flour 10
Cellulase 5 wheat-flours 10 polyphenoloxidase 3.
4. described method by the withered middle extraction tea polysaccharide of tea according to claim 1 is characterized in that described mixed enzyme is to be mixed by the raw material with following weight part:
Saccharifying enzyme 8 hydrolysising proteases 6 flour 8
Cellulase 4 wheat-flours 12 polyphenoloxidase 4.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107349240A (en) * | 2017-06-23 | 2017-11-17 | 百色学院 | A kind of extracting method of mangrove bark polyphenol |
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| CN101942355A (en) * | 2010-08-20 | 2011-01-12 | 上海师范大学 | Comprehensive extraction method for extracting tea seed oil, tea saponin and tea seed polysaccharide from tea seeds or camellia seeds |
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| JPH01304101A (en) * | 1988-06-01 | 1989-12-07 | Riide Chem Kk | Preparation of conjugated polysaccharide |
| KR100602841B1 (en) * | 2005-02-28 | 2006-07-19 | 고려대학교 산학협력단 | Acidic polysaccharide with cell-binding inhibition activity isolated from green tea and its composition for preventing and treating gastrointestinal diseases |
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| CN107349240A (en) * | 2017-06-23 | 2017-11-17 | 百色学院 | A kind of extracting method of mangrove bark polyphenol |
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