CN103059156B - Method for extracting tea polysaccharide from tea seed cake - Google Patents
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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 in tea is withered the method for extraction tea polysaccharide.
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 Tea Polysaccharides mixture, referred to as Tea Polysaccharides or tea polysaccharide.Its protein part is mainly comprised of approximately 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, 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, discovered in recent years tea polysaccharide also has the effect for the treatment of diabetes.From 20 century 70s, launch the research of polysaccharide to begin, develop very fast.Due to the diversified bioactive functions of polysaccharide and at functional food be widely used clinically, the explo iting and researching of polysaccharide Biological resources is become increasingly active, become the study hotspot of natural drug, biological chemistry, life science.Process in advance: tea polysaccharide main source is the dry tea of finished product.By tealeaves is pulverized, at a certain temperature, first use acidity, neutrality or alkalescent water lixiviate, then by precipitate and separate or column chromatography for separation, prepare crude product.At present, the preparation of tea polysaccharide is mainly to extract in tealeaves or 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 by 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 can form water-fast salt with tea polysaccharide as Cetyltrimethyl Ammonium Bromide (CTAB), hexadecyl pyridine hydrochloride (CPC), with this tea polysaccharide and other component separating, recycling dialysis method obtains compared with sterling.
The 3rd, resin method: tea polysaccharide is a kind of acidic polysaccharose, and acidic polysaccharose generally can be adsorbed by associated resin, as DEAE-fibre resin.Then by different elutriants, tea polysaccharide and other compositions can be separated.
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 ", that to take degreasing tea seed grouts, tea-seed grouts be 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 obtained thick tea saponin and tea seed polysaccharide by water extract-alcohol precipitation or alcohol extracting water extraction, clear liquid after water extraction or after alcohol extracting is through macroporous adsorption resin chromatography column purification, resin absorption thing is first with alkali, the ethanol elution that increases gradually by 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 another active result tea polysaccharide simultaneously.Chinese patent (number of patent application is 200710191396.2) " extracting the method for tea polysaccharide from camellia seed meal " and for example, be by camellia seed meal in 10kg than the ratio of 100-200 premium on currency, in water, soak, then heat and stir simultaneously, filter while hot, liquid after filtering is continued to heating, make liquid concentration to 1/4 volume, the yeast adding in concentrated solution, remain at 25-32 ℃ of temperature, vibration fermentation, after fermentation time 0.5-3 days, stop vibration, make its precipitation, getting supernatant liquid adds the ethanol of 3 times of clear liquid volumes to stir, then put into refrigerator freezing, refrigerating fulid is added to water makes it dissolve layering and precipitating, get supernatant liquid, by the spray-dried tea polysaccharide that obtains of supernatant liquid, this invention has solved the method for extracting tea polysaccharide at present 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 advantages that DNA purity is higher.
For realizing its object, the technical solution used in the present invention be invention a kind of in tea is withered the method for extraction tea polysaccharide, 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, drop in fermenting container, then add the clear water that accounts for the withered particle weight of tea 30-50%, stirs evenly, and becomes the withered slurry of tea, standby;
B, mixed enzyme preparation:
By the raw material of 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, standby;
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 standby tea, add the mixed enzyme that accounts for the withered particle weight of tea 6-10%, heat to 20-40 ℃, stir evenly, encapsulation process 4-6 days, becomes enzyme and process the withered slurry of tea, standby;
D, squeezing: to standby enzyme, process in the withered slurry of tea, 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 enzyme treatment solution, standby;
E, primary drying: standby enzyme treatment solution is heated to 70-100 ℃, and 6-10 hour is processed in insulation, then adds edible ethanol, makes the volumetric concentration of enzyme treatment solution alcohol reach 50%, then, at 4-12 ℃ of refrigeration 10-15 hour, filter, and collects mixing liquid; Again by mixing liquid at 40-90 ℃ of vacuum concentration, concentration time is 2-3.5 hour, obtains concentrated solution; Spray-dried again, obtain dry powder, standby;
F, wash-out: to add in standby dry powder account for dry powder weight 1-2 doubly, the volumetric concentration polar organic solvent that is 95%, stir evenly rear standing 2-4 hour, filter, isolate polar organic solvent, so process twice; Then, then to recrement add account for recrement weight 1-2 doubly, volumetric concentration is the low polar organic solvent of more than 95% (or being neat solvent), stirs evenly rear standing 1-2 hour, filtration, so processes three times, collection and treatment slag is standby;
Described polar organic solvent is a kind of in ethanol, methyl alcohol, ethyl acetate, propyl carbinol or two or more, 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: to adding in standby processing slag to account for, process the methyl alcohol that slag weight 20-40%, volumetric concentration are 95%, then add and account for the saturated lead sulfate aqueous solution of processing slag weight 5-12%, process 8-12 hour; And then add to account for and process the ferric sulfate aqueous solution that slag weight 3-5%, concentration are 10-25%, process 0.8-1.2 hour, filter, isolate liquid, collecting precipitation, standby;
H, alkali are molten: in standby precipitation, add precipitation weight 0.5-1.5 doubly, the concentration weak caustic solution that is 1.5-2.4%, dissolve 20-40min; Then, then add 3-5 doubly to precipitate 60-85 ℃ of warm water of weight, soak and extract, filter, collect the aqueous solution, standby;
Described weak caustic solution is sodium soap or sodium bicarbonate or sodium carbonate solution;
I, centrifugal: the standby aqueous solution is centrifugal through the supercentrifuge of turn/min of 5000-10000, obtain centrifugate, standby;
J, mistake post I: by standby centrifugate, through mixing column filtration treatment, coutroi velocity is 50-70L/h, collects filtrate, standby;
K, mistake post II: standby filtrate is crossed to post through polar macroporous resin and process, coutroi velocity is 15-25L/h, collects post liquid, standby;
The model of described polar macroporous resin is D301;
L, sediment: standby crossed to post liquid at 70-100 ℃ vacuum concentration to 1/3, became 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, then add volume be post concentrated solution 1-2 doubly, with water-insoluble organic solvent, stir process 20-36 hour, standing, layering, collect lower floor's insolubles, standby;
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: standby insolubles is dissolved by 1-3 normal-temperature water doubly, filter, collect lysate, standby;
N, redrying: by standby lysate, spraying is dry, obtains tea polysaccharide finished product;
Mixing column in J step is two or more mixture in neutral gac or quartz sand or diatomite or polymeric adsorbent.
The technical scheme one of its optimization: described mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 5 hydrolysising protease 8 flour 5
Cellulase 3 wheat-flour 15 polyphenoloxidase 5.
The technical scheme two of its optimization: described mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 10 hydrolysising protease 4 flour 10
Cellulase 5 wheat-flour 10 polyphenoloxidase 3.
The technical scheme three of its optimization: described mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 8 hydrolysising protease 6 flour 8
Cellulase 4 wheat-flour 12 polyphenoloxidase 4.
Of the present invention in tea is withered the method for extraction tea polysaccharide, to adopt the tea squeezing after 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, the step such as post II, sediment, dissolving, redrying excessively, contained tea polysaccharide during tea is withered has been proposed effectively, and removed the impurity in tea polysaccharide finished product, thereby the tea polysaccharide content in its finished product reaches more than 95%, purity is higher.Meanwhile, by the extraction of tea polysaccharide, expand the withered Application Areas of tea, improved the value of the withered utilization of tea.
Table 1 is below that the tea polysaccharide of extracting according to the inventive method contrasts with the detection data 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
From data in table above, can find out, the tea polysaccharide of extracting according to the inventive method, 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.Explanation is below the mode exemplifying, but protection scope of the present invention is not limited to this.
Of the present invention in tea is withered the method for extraction tea polysaccharide, mixed enzyme used is by the raw material of following weight part is mixed:
Saccharifying enzyme 5-10 hydrolysising protease 4-8 flour 5-10
Cellulase 3-5 wheat-flour 10-15 polyphenoloxidase 3-5.
Embodiment mono-
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, drop in fermenting container, then add the clear water that accounts for the withered particle weight 40% of tea, stirs evenly, and becomes the withered slurry of tea, standby;
B, mixed enzyme preparation:
By the raw material of following weight part:
Saccharifying enzyme 5 hydrolysising protease 8 flour 5
Cellulase 3 wheat-flour 15 polyphenoloxidase 5
Mix, become mixed enzyme, standby;
Saccharifying enzyme used, 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 standby tea, 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 process the withered slurry of tea, standby;
D, squeezing: to standby enzyme, process in the withered slurry of tea, 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 enzyme treatment solution, standby;
E, primary drying: standby enzyme treatment solution is heated to 85 ℃, and insulation is processed 8 hours, then adds edible ethanol, makes the volumetric concentration of enzyme treatment solution alcohol reach 50%, then, 6 ℃ of refrigerations 12 hours, filter, and collects mixing liquid; Again by mixing liquid at 60 ℃ of vacuum concentration, concentration time is 3 hours, obtains concentrated solution (be concentrated into 1/6 or density be 1.2); Spray-dried again, obtain dry powder, standby;
F, wash-out: to add in standby dry powder account for dry powder weight 1-2 doubly, the volumetric concentration polar organic solvent that is 95%, stir evenly rear standing 2-4 hour, filter, isolate polar organic solvent, so process twice; Then, then to recrement add account for recrement weight 1-2 doubly, volumetric concentration is the low polar organic solvent of more than 95% (or being neat solvent), stirs evenly rear standing 1-2 hour, filtration, so processes three times, collection and treatment slag is standby;
To adding in standby dry powder, account for the ethanol that 1.5 times of dry powder weight, volumetric concentration are 95%, stir evenly latter standing 3 hours, filter, isolate ethanol, so process twice; Then, then add and account for the normal hexane that 1.5 times of recrement weight, volumetric concentration are 95% to recrement, stir evenly rear standing 1-2 hour, filter, so process three times, collection and treatment slag, standby;
G, precipitation: to adding in standby processing slag to account for, process the methyl alcohol that slag weight 30%, volumetric concentration are 95%, then add and account for the saturated lead sulfate aqueous solution of processing slag weight 8%, process 10 hours; And then add to account for and process the ferric sulfate aqueous solution that slag weight 4%, concentration are 20%, to process 1 hour, filtration, isolates liquid, and collecting precipitation is standby;
H, alkali are molten: in standby precipitation, add the sodium hydrogen carbonate solution that 1 times of weight of precipitation, concentration are 2.0%, dissolve 30min; Then, then add 4 times of 70 ℃ of warm water that precipitate weight, soak and extract, filter, collect the aqueous solution, standby;
I, centrifugal: the standby aqueous solution is centrifugal through the supercentrifuge of turn/min of 5000-10000, obtain centrifugate, standby;
J, mistake post I: by standby centrifugate, through mixing column (adopting the mixture of neutral gac and quartz sand) filtration treatment, coutroi velocity is 60L/h, collects filtrate, standby; Post used adopts stainless steel column, long 8m, aperture 0.8m;
K, mistake post II: standby filtrate is crossed to post through polar macroporous resin (model is D301) and process, coutroi velocity is 20L/h, collects post liquid, standby;
L, sediment: standby crossed to post liquid at 85 ℃ of vacuum concentration to 1/3, became post concentrated solution, then add gradually ethanol, till while allowing the concentration of ethanol reach 80%, mix thoroughly, then to add volume be the ethyl acetate of 1.5 times of post concentrated solutions, stir process 28 hours, standing, layering, collect lower floor's insolubles, standby;
M, dissolving: standby insolubles is dissolved by the normal-temperature water of 2 times, filter, collect lysate, standby;
N, redrying: by standby lysate, spraying is dry, obtains tea polysaccharide finished product, and tea polysaccharide content wherein reaches 85.1%.
Embodiment bis-
The present embodiment and embodiment mono-are basic identical, and its difference is:
Mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 10 hydrolysising protease 4 flour 10
Cellulase 5 wheat-flour 10 polyphenoloxidase 3;
Mixing column used is the mixture of neutral gac and quartz sand.
The tea polysaccharide finished product extracting, its content is 85.2%.
Embodiment tri-
The present embodiment and embodiment mono-are basic identical, and its difference is:
Mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 8 hydrolysising protease 6 flour 8
Cellulase 4 wheat-flour 12 polyphenoloxidase 4;
Mixing column used is the mixture of diatomite and polymeric adsorbent.
The tea polysaccharide finished product extracting, its content is 85.3%.
Of the present invention in tea is withered the method for extraction tea polysaccharide, 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, the plant that also can be used for containing tea polysaccharide from other, extract tea polysaccharide.
Claims (4)
1. a method for extraction tea polysaccharide in tea is withered, 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, drop in fermenting container, then add the clear water that accounts for the withered particle weight of tea 30-50%, stirs evenly, and becomes the withered slurry of tea, standby;
B, mixed enzyme preparation:
By the raw material of 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, standby;
C, enzyme are processed: in the withered slurry of standby tea, add the mixed enzyme that accounts for the withered particle weight of tea 6-10%, heat to 20-40 ℃, stir evenly, encapsulation process 4-6 days, becomes enzyme and process the withered slurry of tea, standby;
D, squeezing: to standby enzyme, process in the withered slurry of tea, 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 enzyme treatment solution, standby;
E, primary drying: standby enzyme treatment solution is heated to 70-100 ℃, and 6-10 hour is processed in insulation, then adds edible ethanol, makes the volumetric concentration of enzyme treatment solution alcohol reach 50%, then, at 4-12 ℃ of refrigeration 10-15 hour, filter, and collects mixing liquid; Again by mixing liquid at 40-90 ℃ of vacuum concentration, concentration time is 2-3.5 hour, obtains concentrated solution; Spray-dried again, obtain dry powder, standby;
F, wash-out: to add in standby dry powder account for dry powder weight 1-2 doubly, the volumetric concentration polar organic solvent that is 95%, stir evenly rear standing 2-4 hour, filter, isolate polar organic solvent, so process twice; Then, then to recrement add account for recrement weight 1-2 doubly, volumetric concentration is more than 95% normal hexane or sherwood oil or both mixtures, stirs evenly rear standing 1-2 hour, filters, so process three times, collection and treatment slag, standby;
G, precipitation: to adding in standby processing slag to account for, process the methyl alcohol that slag weight 20-40%, volumetric concentration are 95%, then add and account for the saturated lead sulfate aqueous solution of processing slag weight 5-12%, process 8-12 hour; And then add to account for and process the ferric sulfate aqueous solution that slag weight 3-5%, concentration are 10-25%, process 0.8-1.2 hour, filter, isolate liquid, collecting precipitation, standby;
H, alkali are molten: in standby precipitation, add precipitation weight 0.5-1.5 doubly, the concentration weak caustic solution that is 1.5-2.4%, dissolve 20-40min; Then, then add 3-5 doubly to precipitate 60-85 ℃ of warm water of weight, soak and extract, filter, collect the aqueous solution, standby;
I, centrifugal: the standby aqueous solution is centrifugal through the supercentrifuge of turn/min of 5000-10000, obtain centrifugate, standby;
J, mistake post I: by standby centrifugate, through mixing column filtration treatment, coutroi velocity is 50-70L/h, collects filtrate, standby;
K, mistake post II: standby filtrate is crossed to post through polar macroporous resin and process, coutroi velocity is 15-25L/h, collects post liquid, standby;
L, sediment: standby crossed to post liquid at 70-100 ℃ vacuum concentration to 1/3, became 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, then add volume be post concentrated solution 1-2 doubly, with water-insoluble organic solvent, stir process 20-36 hour, standing, layering, collect lower floor's insolubles, standby;
M, dissolving: standby insolubles is dissolved by 1-3 normal-temperature water doubly, filter, collect lysate, standby;
N, redrying: by standby lysate, spraying is dry, obtains tea polysaccharide finished product;
Mixing column in J step is two or more mixture in neutral gac or quartz sand or diatomite or polymeric adsorbent.
2. the method for extraction tea polysaccharide in tea is withered according to claim 1, is characterized in that described mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 5 hydrolysising protease 8 flour 5
Cellulase 3 wheat-flour 15 polyphenoloxidase 5.
3. the method for extraction tea polysaccharide in tea is withered according to claim 1, is characterized in that described mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 10 hydrolysising protease 4 flour 10
Cellulase 5 wheat-flour 10 polyphenoloxidase 3.
4. the method for extraction tea polysaccharide in tea is withered according to claim 1, is characterized in that described mixed enzyme is by the raw material of following weight part is mixed:
Saccharifying enzyme 8 hydrolysising protease 6 flour 8
Cellulase 4 wheat-flour 12 polyphenoloxidase 4.
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| CN101016344A (en) * | 2007-02-13 | 2007-08-15 | 上海师范大学 | Process of preparing high purity tea polysaccharide |
| CN101508690A (en) * | 2009-03-17 | 2009-08-19 | 林峰 | Novel methods for simultaneously extracting tea polyphenol, tea polysaccharide and caffeinum from tea |
| CN101544998A (en) * | 2008-07-01 | 2009-09-30 | 南昌大学 | Separation and purification preparation method and antineoplastic activity of tea polysaccharide |
| CN101575628A (en) * | 2008-07-01 | 2009-11-11 | 南昌大学 | Method for preparing tea polysaccharide through separation and purification and structural identification thereof |
| 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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|---|---|---|---|---|
| CN101016344A (en) * | 2007-02-13 | 2007-08-15 | 上海师范大学 | Process of preparing high purity tea polysaccharide |
| CN101544998A (en) * | 2008-07-01 | 2009-09-30 | 南昌大学 | Separation and purification preparation method and antineoplastic activity of tea polysaccharide |
| CN101575628A (en) * | 2008-07-01 | 2009-11-11 | 南昌大学 | Method for preparing tea polysaccharide through separation and purification and structural identification thereof |
| CN101508690A (en) * | 2009-03-17 | 2009-08-19 | 林峰 | Novel methods for simultaneously extracting tea polyphenol, tea polysaccharide and caffeinum from tea |
| 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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| JP平1-304101A 1989.12.07 |
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