CN101181052B - Method for preparing coriolan - Google Patents
Method for preparing coriolan Download PDFInfo
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- CN101181052B CN101181052B CN2007100324759A CN200710032475A CN101181052B CN 101181052 B CN101181052 B CN 101181052B CN 2007100324759 A CN2007100324759 A CN 2007100324759A CN 200710032475 A CN200710032475 A CN 200710032475A CN 101181052 B CN101181052 B CN 101181052B
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Abstract
The invention relates to a novel preparation method of coriolus versicolor polysaccharide (CVPS), which is characterized in that the mycelium of coriolus versicolor is immerged in hot water based on a mass ratio of 1:4, extraction and filtration are made repeatedly for twice, then water extract is combined and concentrated to one third to one fifth of the original volume; ethanol is added and kept static for sedimentation and then filtered; the sediment is dried to prepare crude amylase to be dissolved with hot water and then added with chloroform and n-butyl alcohol and surged fiercely till jello is generated; centrifugal separation is carried out to gather supernatant fluid; pH is adjusted to be equal to 9.0 by NaOH, then H2O2 is dripped into the water bath with the temperature of 50 DEG C and the pH is kept between 8.0 to 9.0; the water bath lasts for 4h and the reaction stops and ethanol is added slowly; after that, the supernatant fluid is put into a refrigerator for the night and sediment is gathered and dried on next day to get product. The invention can obviously promote the yield of the CVPS, remarkably shorten operation time, reduce the consumption of energy and ensure that the CVPS with relatively higher activity for batch production with less production cost is produced, thus resulting in the commercialization of the CVPS on a larger industrialization degree.
Description
Technical field
The present invention relates to a kind of preparation method of new coriolan.
Background technology
Rainbow conk (Coriolus versicolor) is the Chinese herbal medicine resource of China's preciousness, in the prepared from coriolus versicolor mycelium and external all can produce coriolan (Coriolus versciolor polysaccharides, CVP), this is a kind of proteinaceous Polysaccharide-Peptide complex, and main component is with β (1,4), β (1,3) or β (1,4), β (1,6) is main glucan complex, on β (1 is arranged, 3) or β (1,6) branch of combination, polyoses content 30%～60% is in conjunction with protein 15 %.
Coriolan energy immune cell activated, improve body's immunity, obviously improve the intensity of body inflammatory reaction, have antitumor, antiulcer, reducing blood lipid, multiple biologically active such as protect the liver, function with antibacterium, mould and virus, can stimulate macrophage to transform, increase the interleukin 1 in the blood, the effect of interferon gamma and TNF, can suppress cancer cell generates, be safe, the human or animal's that biologically active is strong immunopotentiator more can be used as the additive of healthy food.
The method of coriolan extraction at present mainly is divided into water extract method, sour formulation, alkaline extraction etc. by the difference of extracting solvent, compare difference with water extract method little though sour formulation and alkaline extraction extract the highest yield of coriolan, but contaminated environment seriously, and the flooding rule has stronger advantage at aspects such as production technology, production cost, Product Safety and environmental protection.
The method that present water extract method often adopts heating to boil manyzoned polypore sporophore is extracted coriolan, this process need 5～10h only, even the longer time, the leaching liquor large usage quantity, sometimes up to 30 times, in leaching process, will consume a large amount of energy, and the yield of coriolan has only about 7.81%.
Summary of the invention
The objective of the invention is to develop a kind of new preparation method that can obtain the coriolan of high yield high-load.
We obtain thick polysaccharide with precipitation with alcohol again by prepared from coriolus versicolor mycelium is immersed lixiviate at least twice in 4 times of hot water, remove albumen again, decolouring, drying, the yield of the coriolan that obtains are at least more than 8%, polyoses content after the purification is more than 40%, thereby realized purpose of the present invention.
The preparation method of a kind of coriolan of the present invention comprises the steps:
(1) prepared from coriolus versicolor mycelium was pressed mass ratio 1: 4, immersion has added mass fraction 2.0%Na
2CO
390 ℃ of hot water in, lixiviate 1～2h, lixiviate is filtered 2～3 times repeatedly, merges leaching liquor then and is concentrated into 1/5～1/3 of original volume;
(2) adding volume in the concentrated extracting solution that obtains of step (1) is the ethanol of 3～4 times of concentrated extracting solutions, filters after staticly settling 24h, and 25 ℃ of oven dry of precipitation part obtain thick polysaccharide;
(3) the thick polysaccharide that obtains of step (2) adds 1/5 chloroform of its aqueous solution volume and the n-butanol of chloroform volume 1/5 with 70 ℃ of hot water dissolvings, and thermal agitation is to generating jello, and supernatant is collected in centrifugation, removes sub-cloud albuminate and chloroform;
(4) supernatant that step (3) is obtained is modulated to the aqueous solution that contains thick polysaccharide 5%, is transferred to pH=9.0 with NaOH, and in 50 ℃ of water-baths, splashing into volume is 15% of thick polysaccharide solution, and mass fraction is 15% H
2O
2Solution, keeping pH is 8.0～9.0, continues cessation reaction behind the water-bath 4h, slowly adds mass fraction 95% ethanol, puts refrigerator overnight, next day collecting precipitation, drying obtains product.
The described prepared from coriolus versicolor mycelium of step (1) can obtain by filtering rainbow conk mycelia zymotic fluid, preferably immerses 2h in the hot water, adds Na in the described hot water
2CO
3, can prevent the polysaccharide degraded.
The described duration of oscillation of step (3) is 20～30min preferably, and described centrifugation condition is rotating speed 4000r/min preferably, continues 15min.
The described drying of step (4) preferably adopts vacuum freeze drying.
Preparation method of the present invention can significantly improve the yield of coriolan, and obviously shorten the operating time, reduce the consumption of the energy, under less production cost, can guarantee to produce in enormous quantities the active higher coriolan of acquisition, thereby make coriolan on bigger industrialization degree, realize commercialization.
Description of drawings
Fig. 1: the ultra-violet absorption spectrum of prepared from coriolus versicolor mycelium extract.
Fig. 2: glucose content calibration curve
The specific embodiment
Embodiment 1:
Take by weighing prepared from coriolus versicolor mycelium net weight 122.13g (oven-dried weight 16.22g), pulverize behind the adding 125mL pure water, add adding water to 350mL, add 7gNa then
2CO
3, behind 90 ℃ of water-bath 2h, lixiviate is filtered 2 times repeatedly, gets leaching liquor then and is concentrated into 1/5 of original volume.95% ethanol that the concentrate 200mL that obtains is added 3 times of volumes precipitates, and filters after leaving standstill 24h, and the precipitation partially desiccated obtains thick polysaccharide.Add 70 ℃ of hot water 200mL and dissolve thick polysaccharide, add the 40mL chloroform, the 8mL n-butanol, thermal agitation 20min generates jello, 4000r/min centrifugal filtration 15min on centrifuge, collect supernatant, remove sub-cloud albuminate and upper strata chloroform, obtain solution 130mL.Regulate pH value to 9.0 with NaOH, add 15%H
2O
220mL, and then 50 ℃ of water-bath 4h, and slowly add 95% ethanol 150mL.4 ℃ of refrigerator overnight, next day 4000r/min, 20min centrifugal filtration, collecting precipitation, vacuum drying obtains white powder 1.432g, yield 8.83%.
Embodiment 2: the ultra-violet absorption spectrum of the product that embodiment 1 obtains is measured
Ultraviolet scanner HITACHI (Japan) carries out the UV scanning of 190～360nm to sample aqueous solution, and to determine whether the being protein combination polysaccharide, measurement result is seen accompanying drawing 1, and coriolan is a kind of proteinaceous Polysaccharide-Peptide complex.Do not have absorption from the graph as can be seen at the 260nm place, a shoulder shape peak is arranged about 270-280nm, this is the characteristic absorption of protein combination polysaccharide.
Embodiment 3: the coriolan assay of the product that embodiment 1 obtains
Accurately take by weighing standard specimen glucose 20mg in the 500mL volumetric flask, add water to scale, draw 0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6 respectively, 1.8mL, mend to 2.0mL with water respectively, add 5% phenol 1.0mL and concentrated sulfuric acid 5.0mL then, leave standstill 10min, shake up, 30 ℃ of water-bath 20min.The back is in 490nm photometry density value (OD), and 2.0mL water is blank.Abscissa is a concentration of glucose, and ordinate is the OD value, paints canonical plotting (seeing accompanying drawing 2), carries out linear recurrence, gets regression equation: y=0.0146x+0.0103, R
2=0.9984.Draw sample liquid 1.0ml, operating procedure is the same, and triplicate is surveyed the OD value and is respectively 0.2983,0.2901,0.2956, gets its mean value, looks into calibration curve.The correction coefficient of coriolan and glucose content is 2.5, and as calculated, the back polyoses content of purifying is 48.69%.
Claims (2)
1. the preparation method of a coriolan comprises the steps:
(1) prepared from coriolus versicolor mycelium was pressed mass ratio 1: 4, immersion has added mass fraction 2.0%Na
2CO
390 ℃ of hot water in, lixiviate 1～2h, lixiviate is filtered 2～3 times repeatedly, merges leaching liquor then and is concentrated into 1/5～1/3 of original volume;
(2) adding volume in the concentrated extracting solution that obtains of step (1) is the ethanol of 3～4 times of concentrated extracting solutions, filters after staticly settling 24h, and 25 ℃ of oven dry of precipitation part obtain thick polysaccharide;
(3) the thick polysaccharide that obtains of step (2) adds 1/5 chloroform of its aqueous solution volume and the n-butanol of chloroform volume 1/5 with 70 ℃ of hot water dissolvings, and thermal agitation is to generating jello, and supernatant is collected in centrifugation, removes sub-cloud albuminate and chloroform;
(4) supernatant that step (3) is obtained is modulated to the aqueous solution that contains thick polysaccharide 5%, is transferred to pH=9.0 with NaOH, and in 50 ℃ of water-baths, splashing into volume is 15% of thick polysaccharide solution, and mass fraction is 15% H
2O
2Solution, keeping pH is 8.0～9.0, continues cessation reaction behind the water-bath 4h, slowly adds mass fraction 95% ethanol, puts refrigerator overnight, next day collecting precipitation, drying obtains product.
2. the preparation method of a kind of coriolan according to claim 1, the time that it is characterized in that lixiviate in step (1) hot water is 2h, the described duration of oscillation of step (3) is 20～30min, described centrifugation condition is rotating speed 4000r/min, continue 15min, the described dry vacuum freeze drying that adopts of step (4).
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| CN2007100324759A CN101181052B (en) | 2007-12-14 | 2007-12-14 | Method for preparing coriolan |
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| CN2007100324759A CN101181052B (en) | 2007-12-14 | 2007-12-14 | Method for preparing coriolan |
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| CN101181052B true CN101181052B (en) | 2010-12-01 |
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102078600B (en) * | 2010-12-24 | 2014-03-26 | 区士衡 | Anti-cancer compound ganoderma composition, application thereof and pharmaceutical composition containing same |
| CN103211066A (en) * | 2013-05-10 | 2013-07-24 | 北京绿源求证科技发展有限责任公司 | Food-grade health tea granules capable of improving sleeping |
| CN105325821A (en) * | 2014-06-27 | 2016-02-17 | 北大方正集团有限公司 | Healthy functional beverage and preparation method thereof |
| CN104387486A (en) * | 2014-11-11 | 2015-03-04 | 罗宇 | High-yield preparation method of coriolus versicolor polysaccharide |
| CN104382929A (en) * | 2014-11-25 | 2015-03-04 | 高秀媛 | Medicine for removing blood garbage and vivotoxin and preparation method thereof |
| CN104382000A (en) * | 2014-12-04 | 2015-03-04 | 李振铭 | Coriolus versicolor, tremella aurantialba and poria cocos polysaccharide and preparation process thereof |
| CN104473284A (en) * | 2014-12-31 | 2015-04-01 | 华南师范大学 | Coriolus versicolor polysaccharide functional drink and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1047805A (en) * | 1990-04-20 | 1990-12-19 | 长春制药厂 | A kind of polysaccharide-k extracting new process |
| US6087335A (en) * | 1992-11-30 | 2000-07-11 | Yang; Mable M. P. | RNase-CV (Coriolus versicolor) |
| US20070104727A1 (en) * | 2005-11-08 | 2007-05-10 | Vita Green Health Products Co., Ltd | Herbal powder extracts and methods of preparing and using the same |
-
2007
- 2007-12-14 CN CN2007100324759A patent/CN101181052B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1047805A (en) * | 1990-04-20 | 1990-12-19 | 长春制药厂 | A kind of polysaccharide-k extracting new process |
| US6087335A (en) * | 1992-11-30 | 2000-07-11 | Yang; Mable M. P. | RNase-CV (Coriolus versicolor) |
| US20070104727A1 (en) * | 2005-11-08 | 2007-05-10 | Vita Green Health Products Co., Ltd | Herbal powder extracts and methods of preparing and using the same |
Non-Patent Citations (2)
| Title |
|---|
| 张志军等.灵芝多糖提取———水浸提条件的研究.《天津农学院学报》.2005,第12 卷(第1期),12-15. * |
| 田光辉.香菇多糖提取工艺的优化.《延安大学学报(自然科学版)》.2002,第21卷(第1期),46-47. * |
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