CN1144847A - Method for extracting asponin composition from traditional Chinese medicinal material with enzymic processing - Google Patents
Method for extracting asponin composition from traditional Chinese medicinal material with enzymic processing Download PDFInfo
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- CN1144847A CN1144847A CN96115151A CN96115151A CN1144847A CN 1144847 A CN1144847 A CN 1144847A CN 96115151 A CN96115151 A CN 96115151A CN 96115151 A CN96115151 A CN 96115151A CN 1144847 A CN1144847 A CN 1144847A
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Abstract
An enzyme process for extracting saponin from Chinese-medicinal materials includes such steps as immersing the Chinese-medicinal materials in water, mixing endoglycosidases or the enzymes not containing exoglycosidase components with the immersed liquid, reaction, immersing the materials to be extracted in water, alcohol or methanol to extract saponin, and features high extracting rate of saponin (20-150%), no damage to saponin structure, and also extracting other soluble matters with higher extracting rate.
Description
The present invention relates to a kind of method of extracting saponin composition in the Chinese medicinal materials, particularly adopt enzyme to handle the method for extracting saponin composition in the Chinese medicinal materials.
Genseng is universally acknowledged rare medicinal herbs, and it mainly contains effective constituent is ginsenoside, kind surplus the ginsenoside kind of having known has 30.Be mixed with spirit of ginseng, panax ginseng extractum, Radix Ginseng tea (aqueous, solid shape) after people's effective ingredient～ginsenoside leaching extraction often, also have and nourishing drinks such as other drug blended health tea, drinks genseng.
Prior art is for the extraction of ginsenoside, all is to adopt lixiviation process, that is: water, alcohol or other organic solvent genseng after to fragmentation soaks, thereby leaches, extraction ginsenoside composition.As disclosed water, ethanol lixiviation process in " Chinese Pharmaceutical Journal " (27 (5), 291～293,1992), be in the ginseng powder, to add 40% ethanol to leach, filter; Adding boiling water in the slag again leaches, filters.Slag can repeat to extract, and filtrate merges, and concentrates, and adds ethanol reconcentration, ether defatting, n-butanol extraction, and the pressure reducing and steaming propyl carbinol can get saponin.And for example the Shibata method of Japan's special permission communique (91109,1978) announcement also is with alcohol genseng to be leached, extract saponin, ether defatting, n-butanol extraction, and the pressure reducing and steaming propyl carbinol can get saponin.
Above-mentioned lixiviation process exists a disadvantage: no matter adopt what solvent and what condition to leach, 20～60% above saponin effective ingredients will still remain in by among the ginseng residue after leaching.In other words, prior art is too low to the extraction yield of saponin, has wasted the resource of genseng one class rare medicinal herbs.
The object of the invention is: overcome the deficiency of prior art, provide a kind of to being that the Chinese medicinal materials that mainly contains effective constituent carries out the method that enzyme is handled its saponin composition of extraction with saponin, be intended to increase substantially extraction yield.
Why low to the saponin extraction yield lixiviation process of prior art is, this is that be good for glucosides with other polysaccharide or biomacromolecule at the sugared position of a big chunk saponin or ester is strong combines because contain in the genseng, saponin part is adsorbed on polysaccharide such as genseng fiber, pectin, starch and other biomacromolecule securely, thereby can not extract this part saponin composition with water, alcohol or the organic solvent immersion of pure physics.Hundreds of kinds such as Radix Codonopsis, balloonflower root, the tuber of dwarf lilyturf, the root of straight ladybell, Huang Ling are the Chinese medicinal materials that mainly contains effective constituent with saponin, and are all similar to above-mentioned genseng situation.
Basic thought of the present invention is at above-mentioned principle, utilize the endoglycosidase class and other enzyme that does not contain exoglycosidase that do not destroy saponin to handle Chinese medicinal materialss such as genseng: the one, cutting saponin sugar and other polysaccharide institute bonded valence bond, strong etc. as glucosides, saponin is come off; The 2nd, the biomacromolecule of hydrolysis absorption saponin makes it to come off.And then thereby the row leaching increases substantially the saponin extraction yield.
Implementation of the present invention is: enzyme is handled the method for extracting saponin composition in the Chinese medicinal materials, it is characterized in that:
(1), extract is soaked,
(2), get endoglycosidase class or other and do not contain the enzyme of exoglycosidase composition and extract water logging liquid-phase mixing, reaction,
(3), make leaching agent, adopt the leaching method step that the extract through above-mentioned enzyme treatment step is leached, extracts the saponin composition with water, ethanol or methyl alcohol.
With above-mentioned scheme implementation when of the present invention, after can and soaking water with extract and together heating, be cooled to the temperature that will use enzyme suitable, add corresponding enzyme again.Selected enzyme can be following wherein one or more combination:
(1), amylases: α-Dian Fenmei, beta-amylase, diastatic enzyme,
(2) Mierocrystalline cellulose restriction endonuclease,
(3), the pectin enzyme,
(4), mould Pseudomonas and basidiomycetous liquid koji or solid koji leach liquor enzymes such as Aspergillus, Rhizopus, Mucor, red colouring agent for food, also used as a Chinese medicine, the mixed enzyme of the leach liquor of natural song, the enzyme that the plant seed bud leaches.
In addition, when enforcement is of the present invention, if contain the exoglycosidase class composition of hydrolysis saponin, remove its enzymic activity in the selected enzyme with heating or inhibiting method.
Advantage of the present invention is:
1, the present invention not only is applicable to whole plant materialss of all kinds genseng in the Panax (Panax genus), is applicable to that also all are Chinese medicinal materials (as Radix Codonopsis, balloonflower root, the tuber of dwarf lilyturf, the root of straight ladybell, Huang Ling, Caulis Bauhihiae Championii, the gynostemma pentaphylla) plant that mainly contains effective constituent (as soybean etc.) with saponin.
2, implement the present invention, can make the saponin extraction yield improve more than 20～150% than prior art lixiviation process.
3, adopt the present invention that genseng and other Chinese medicinal materials are carried out the enzyme processing, the extraction that not only improves effective ingredients such as saponin has also improved the extraction yield to other soluble substance (as polysaccharide, polypeptide) simultaneously.
4, adopt the saponin that the present invention extracted, measure, do not find the considerable change (can referring to accompanying drawing) of its composition, illustrate that the enzyme processing does not destroy its saponin structure with silica gel G thin layer chromatography or high pressure liquid chromatographic analysis method.
Fig. 1 extracts the high pressure liquid chromatography figure of saponin with the methyl alcohol lixiviation process for prior art.
Fig. 2 extracts the high pressure liquid chromatography figure of saponin with the ethanol lixiviation process for prior art.
Fig. 3 extracts the high pressure liquid chromatography figure of saponin with Glycosylase processing～ethanol lixiviation process for the present invention.
Fig. 4 extracts the high pressure liquid chromatography figure of saponin with the bent leach liquor enzyme of mould processing～ethanol lixiviation process for the present invention.
More than each figure: instrument, Shimadzu; Chromatographic column, φ 0.9 * 29cm carbohydrate post; Detector, RI; Moving phase, second cyanogen: water=80: 20; Flow velocity, 2ml/ minute; Annotate auspicious amount, 20 μ l.
Below in conjunction with embodiment and accompanying drawing the present invention is done further narration:
Embodiment 1:
Add 200～1000ml water or 0.01M in 40 purpose ginseng powder, 100 grams, the acetate buffer solution of PH5.5 (water addition ratio can be 1.5～200 times), handled 20 minutes for 100 ℃, be cooled to 60 ℃, add the Mierocrystalline cellulose restriction endonuclease, handled two hours at 40～60 ℃, add 95% ethanol, 350～850ml, centrifuging gets filtrate; Add 40% ethanol 500ml in the slag again, leached 2 hours at 80 ℃, centrifuging, filtrate merges (leaching can repeat several times), be concentrated into 1/10 left and right sides volume, add with volume ether defatting (ether defatting repeats 3～6 times), water intaking layer 50ml water-saturated n-butanol extraction saponin ten times in the debris, combining extraction liquid, pressure reducing and steaming propyl carbinol can get the ginsenoside of the above beige of 7～11 grams.
From the identical ginseng powder of 100 grams who handles without enzyme, get the saponin of 4～6 gram same purity only with above-mentioned same procedure.Its ginseng residue handles through enzyme can get 1～3 gram saponin.
Obtaining ginsenoside from aforesaid method, is that standard, Vanillin colour developing colorimetry (Zou Chunjie, Xu Jingda: herbal medicine, 13 (8), 19～23,1982) are measured its content with Re, and error is no more than 1%; Recycle silicon glue G thin layer chromatography 82) measure its content, error is no more than 1%; Recycle silicon glue G thin layer chromatography (Zou Chunjie, Xu Jingda: herbal medicine, 13 (8), 19～23,1982) or high pressure liquid chromatographic analysis method (Nagasawa T.et al:Chem.Pharm.Bull., 28,2059-2064,1980.) measure its each saponin, compare with Fig. 1,2 referring to Fig. 3, do not find the considerable change of its composition, illustrate and do not destroy its saponin structure, method is very successful.
Embodiment 2:
Add 200～2000ml water among 40 purposes, the 100 gram ginseng powders, handled 20 minutes for 100 ℃, be cooled to 50 ℃, add 5～300,000 unit amylase or 1～100,000 unit diastatic enzyme again, stirring reaction 2 hours; Add 95% ethanol to 40% (V/V), 80 ℃ are incubated 2 hours, centrifugal collection leach liquor, its slag leach several times with 40% ethanol again, merge leach liquor and are concentrated into 1/10 volume, use 80ml ether defatting 5～7 times at every turn, collect water with 100ml water-saturated n-butanol extraction 7 times, collect propyl carbinol phase evaporated under reduced pressure, residue is dissolved in 80ml methyl alcohol, filtration again, methanol solution is dripped in the 1000ml acetone, and collecting precipitation, oven dry promptly get 6～9 gram ginsenoside powder of beige.
Handle, can only get as stated above 3～4 gram ginsenoside powder without enzyme.Its ginseng residue extracts saponin and can get about 1～3 gram again through enzyme processing as stated above.
Embodiment 3:
100 grams, 40 purpose ginseng powders add water 400～2000ml, handled 20 minutes for 100 ℃, be cooled to 45～55 ℃, add again the mould meet codex alimentarius (as aspergillus oryzae, head mold ...) nutrient solution or mould solid culture leach liquor enzyme (the redox enzymes activity that does not contain beta-glucosidase, α-xylosidase, α-arabinofuranosidase/xylosidase and α-rhamnosidase and ginsenoside source) handled 1.5～6 hours, again with above-mentioned ethanol or methyl alcohol method extraction saponin.Improved 30～150% than the saponin extraction yield of handling without enzyme liquid.
Obtain ginsenoside from aforesaid method, with Re is standard, Vanillin colour developing colorimetry (Zou Chunjie, Xu Jingda: herbal medicine, 13 (8), 19～23,1982) measure its content, error is no more than 1%, recycle silicon glue G thin layer laminate method (Zou Chunjie, Xu Jingda: herbal medicine, 13 (8), 19～23,1982) or high pressure lipuid chromatography (HPLC) (Nagasawa T.et al:Chem.Pharm.Bull., 28,2059-2064,1980) measure its saponin, referring to Fig. 4 and Fig. 1,2 relatively, do not find the considerable change of its composition, illustrate and do not destroy its saponin structure, method is very successful.
Embodiment 4:
100 gram ginseng powders handle genseng or other Chinese medicine as stated above, not only improve the extraction yield of saponin greatly, and what make whole genseng becomes soluble substance more than 40～60%, handle than enzyme not and improve 100～250%.
Embodiment 5:
Replace genseng with balloonflower root, press the method for embodiment 1 and extract saponin, the kikyosaponin extracted amount that enzyme is handled is to be equivalent to not handle 120～200% of kikyosaponin extracted amount with enzyme; To cross the slag of saponin be raw material not handle Radix Platycodonis extract with enzyme, handles, extracts saponin through enzyme again, and extracted amount can reach 20～100% of former extracted amount again.Extract saponin as stated above with Radix Codonopsis, the tuber of dwarf lilyturf, the root of straight ladybell, Huang Ling, Caulis Bauhihiae Championii, gynostemma pentaphylla, soybean etc., the untreated saponin extraction yield of handling through enzyme of ratio has improved 30～100%.
Claims (4)
1, enzyme is handled the method for extracting saponin composition in the Chinese medicinal materials, it is characterized in that:
(1), extract is soaked,
(2), get endoglycosidase class or other and do not contain the enzyme of exoglycosidase composition and extract water logging liquid-phase mixing, reaction,
(3), make leaching agent, adopt the leaching method step that the extract through above-mentioned enzyme treatment step is leached, extracts the saponin composition with water, ethanol or methyl alcohol.
2, extraction saponin method according to claim 1 is characterized in that: with extract and after soaking water and together heating, be cooled to the temperature that will use enzyme suitable, add corresponding enzyme again.
3, extraction saponin method according to claim 1 and 2, it is characterized in that: selected enzyme can be following wherein one or more combination:
(1), amylases: α-Dian Fenmei, beta-amylase, diastatic enzyme,
(2) Mierocrystalline cellulose restriction endonuclease,
(3), the pectin enzyme,
(4), mould Pseudomonas and basidiomycetous liquid koji or solid koji leach liquor enzymes such as Aspergillus, Rhizopus, Mucor, red colouring agent for food, also used as a Chinese medicine, the mixed enzyme of the leach liquor of natural song, the enzyme that the plant seed bud leaches.
4, according to the described method of claim 3, it is characterized in that: if contain the exoglycosidase class composition of hydrolysis saponin, remove its enzymic activity in the selected enzyme with heating or inhibiting method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN96115151A CN1048283C (en) | 1996-03-12 | 1996-03-12 | Method for extracting asponin composition from traditional Chinese medicinal material with enzymic processing |
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| Application Number | Priority Date | Filing Date | Title |
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| CN96115151A CN1048283C (en) | 1996-03-12 | 1996-03-12 | Method for extracting asponin composition from traditional Chinese medicinal material with enzymic processing |
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| CN1144847A true CN1144847A (en) | 1997-03-12 |
| CN1048283C CN1048283C (en) | 2000-01-12 |
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| CN96115151A Expired - Fee Related CN1048283C (en) | 1996-03-12 | 1996-03-12 | Method for extracting asponin composition from traditional Chinese medicinal material with enzymic processing |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002053722A1 (en) * | 2000-12-29 | 2002-07-11 | Fengxie Jin | Ginsenoside glycosidase which hydrolyzes ginsenoside glycosyl and the use thereof |
| CN1105781C (en) * | 1999-03-17 | 2003-04-16 | 金凤燮 | Method for preparing rare ginsengoside using enzymatic method to modify ginsenoside glycoside |
| CN104523789A (en) * | 2014-12-19 | 2015-04-22 | 吉林玉参医药科技有限公司 | Method for preparing brown ginseng extract with high antitumor activity and application of method |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE1492130B1 (en) * | 1963-05-30 | 1971-08-26 | Shionogi & Co | Process for the production of saponin tetraglycosides from leaves of digitalis plants |
| CN1033278A (en) * | 1987-11-24 | 1989-06-07 | 白求恩医科大学 | Technology for producing ginsenoside re monomer |
| CN1089210A (en) * | 1992-03-21 | 1994-07-13 | 武冈县蜂窝煤机制造厂 | Horizontal hydraulic briquette forming machine |
| CN1080291A (en) * | 1992-06-26 | 1994-01-05 | 郴州科技实业公司 | From tea waste, extract the method for refined saponin |
| JPH06287554A (en) * | 1993-04-01 | 1994-10-11 | Aomori Pref Gov | Production of oxidation-resistant substance from soybean meal or lees of bean curd |
-
1996
- 1996-03-12 CN CN96115151A patent/CN1048283C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1105781C (en) * | 1999-03-17 | 2003-04-16 | 金凤燮 | Method for preparing rare ginsengoside using enzymatic method to modify ginsenoside glycoside |
| WO2002053722A1 (en) * | 2000-12-29 | 2002-07-11 | Fengxie Jin | Ginsenoside glycosidase which hydrolyzes ginsenoside glycosyl and the use thereof |
| US7759101B2 (en) | 2000-12-29 | 2010-07-20 | Fengxie Jin | Ginsenoside glycosidases hydrolyzing ginsenoside sugar moieties and uses thereof |
| CN104523789A (en) * | 2014-12-19 | 2015-04-22 | 吉林玉参医药科技有限公司 | Method for preparing brown ginseng extract with high antitumor activity and application of method |
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| CN1048283C (en) | 2000-01-12 |
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