CN105061628B - The extracting method of coriolan - Google Patents
The extracting method of coriolan Download PDFInfo
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- CN105061628B CN105061628B CN201510542537.5A CN201510542537A CN105061628B CN 105061628 B CN105061628 B CN 105061628B CN 201510542537 A CN201510542537 A CN 201510542537A CN 105061628 B CN105061628 B CN 105061628B
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Abstract
The invention discloses a kind of extracting method of coriolan, comprise the following steps：1) rainbow conk ethanol is extracted, takes filter residue standby；2) by filter residue water extraction, it is standby to collect filtrate；3) by filtrate concentrated extract, ethanol precipitation, centrifuging to precipitate；4) by precipitation, absolute ethyl alcohol, acetone wash respectively successively, precipitation after washing is dissolved with distilled water, cross the hollow-fibre membrane that molecular cut off is 1,000,000, collect concentrate, then concentrate is crossed into the hollow-fibre membrane that molecular cut off is 1,500,000, permeate centrifugation is collected, filtering, takes filtrate standby；5) filtrate is taken to be spray-dried, as coriolan.The coriolan purity of the present invention is high, and color and luster is whiter, and antitumor activity is strong.
Description
Technical field
The invention belongs to technical field of biological extraction, and in particular to the extracting method of coriolan.
Background technology
Coriolan is one of important chemical composition of rainbow conk, has antitumor, Immune-enhancing effect and liver-protective effect.
However, coriolan, because extraction process falls behind, impurity content is high, and color and luster is in orange grey, and moreover, conventional method is extracted
Inhibition rate of tumor cell to coriolan is only 30% or so.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of extracting method of coriolan, the rainbow conk that this method obtains is more
Sugared purity is high, and active anticancer is good.
Technical scheme provided by the invention is the extracting method of coriolan, is comprised the following steps：
1) rainbow conk is crushed, extracted 2~3 times with 6~12 times of 80~95v% of weight alcohol refluxs, flow back 2~3h every time,
Filtering, takes filter residue standby；
2) the water refluxing extraction 2~3 times by filter residue with 6~12 times of weight, 2~3h is extracted every time, filtering, it is standby to collect filtrate
With；
3) medicinal extract that relative density at 60 DEG C is 1.1~1.3 is concentrated the filtrate to, adds ethanol precipitation, centrifuging to precipitate；
4) precipitation is washed respectively with the absolute ethyl alcohol of 2~4 times of weight, acetone successively, by the precipitation after washing with 2~4
The dissolving of times distilled water, the hollow-fibre membrane that molecular cut off is 1,000,000 is crossed, collects concentrate, concentrate is then crossed into retention point
Son amount is 1,500,000 hollow-fibre membrane, collects permeate centrifugation, filtering, takes filtrate standby；
5) filtrate is taken to be spray-dried, as coriolan.
In step 3), ethanol to the ethanol content that 90~95v% is added into medicinal extract is 80~85v%.
In step 4), after the precipitation after washing is dissolved in distilled water, the hollow-fibre membrane that molecular cut off is 1,000,000 is first crossed,
So polysaccharide and oligosaccharide of the molecular weight less than 1,000,000, monose and pigment are as permeate flows out, the concentrate mistake of collection
Molecular cut off is that the polysaccharide of 1,500,000 hollow-fibre membrane, macro-molecular protein and molecular weight higher than 1,500,000 is trapped, and is received
Collect permeate.
The present invention crosses hollow-fibre membrane twice, the removal of impurity and pigment not only can be farthest gone, to ensure that finished product is pure
Degree and color and luster, and the coriolan molecular weight that extraction obtains, between 1,000,000~1,500,000, its antitumor activity is stronger.
Embodiment
The present invention is further elaborated for specific examples below, but not as a limitation of the invention.
Embodiment 1
1) rainbow conk is crushed, extracted 2 times with 6 times of weight 80v% alcohol refluxs, flow back 2h every time, filtering, takes filter residue standby
With；
2) the water refluxing extraction 2 times by filter residue with 6 times of weight, extracts 2h every time, filters, and it is standby to collect filtrate；
3) medicinal extract that relative density at 60 DEG C is 1.1 is concentrated the filtrate to, addition 90v% ethanol is to ethanol into medicinal extract
Content precipitates for 80v%, centrifugation, is precipitated；
4) precipitation is washed respectively with the absolute ethyl alcohol of 2 times of weight, acetone successively, the precipitation after washing is distilled with 2 times
Water dissolves, and crosses the hollow-fibre membrane that molecular cut off is 1,000,000, collects concentrate, concentrate then is crossed into molecular cut off is
1500000 hollow-fibre membrane, permeate centrifugation is collected, filtering, takes filtrate standby；
5) filtrate is taken to be spray-dried, as coriolan.It is 93.6% with phend-sulphuric acid measure polyoses content, color
For white.
Control group
1) rainbow conk is crushed, extracted 2 times with 6 times of weight 80v% alcohol refluxs, flow back 2h every time, filtering, takes filter residue standby
With；
2) the water refluxing extraction 2 times by filter residue with 6 times of weight, extracts 2h every time, filters, and it is standby to collect filtrate；
3) medicinal extract that relative density at 60 DEG C is 1.1 is concentrated the filtrate to, addition 90v% ethanol is to ethanol into medicinal extract
Content precipitates for 80v%, centrifugation, is precipitated；
4) precipitation is washed respectively with the absolute ethyl alcohol of 2 times of weight, acetone successively, the precipitation after washing is distilled with 2 times
Water dissolves, upper D303 types macroreticular resin, is eluted with water to efflux water white transparency, collects efflux, concentrates, spray drying, i.e.,
For coriolan.It is 85.3% with phend-sulphuric acid measure polyoses content, color is orange grey.
Embodiment 2
1) rainbow conk is crushed, extracted 3 times with 12 times of weight 95v% alcohol refluxs, flow back 3h every time, filtering, takes filter residue standby
With；
2) the water refluxing extraction 3 times by filter residue with 12 times of weight, extracts 3h every time, filters, and it is standby to collect filtrate；
3) medicinal extract that relative density at 60 DEG C is 1.3 is concentrated the filtrate to, addition 95v% ethanol is to ethanol into medicinal extract
Content precipitates for 85v%, centrifugation, is precipitated；
4) precipitation is washed respectively with the absolute ethyl alcohol of 4 times of weight, acetone successively, the precipitation after washing is distilled with 4 times
Water dissolves, and crosses the hollow-fibre membrane that molecular cut off is 1,000,000, collects concentrate, concentrate then is crossed into molecular cut off is
1500000 hollow-fibre membrane, permeate centrifugation is collected, filtering, takes filtrate standby；
5) filtrate is taken to be spray-dried, as coriolan.It is 93.8% with phend-sulphuric acid measure polyoses content, color
For white.
Embodiment 3
1) rainbow conk is crushed, extracted 2 times with 10 times of weight 90v% alcohol refluxs, flow back 2h every time, filtering, takes filter residue standby
With；
2) the water refluxing extraction 2 times by filter residue with 10 times of weight, extracts 2h every time, filters, and it is standby to collect filtrate；
3) medicinal extract that relative density at 60 DEG C is 1.2 is concentrated the filtrate to, addition 95v% ethanol is to ethanol into medicinal extract
Content precipitates for 85v%, centrifugation, is precipitated；
4) precipitation is washed respectively with the absolute ethyl alcohol of 3 times of weight, acetone successively, the precipitation after washing is distilled with 3 times
Water dissolves, and crosses the hollow-fibre membrane that molecular cut off is 1,000,000, collects concentrate, concentrate then is crossed into molecular cut off is
1500000 hollow-fibre membrane, permeate centrifugation is collected, filtering, takes filtrate standby；
5) filtrate is taken to be spray-dried, as coriolan.It is 94.1% with phend-sulphuric acid measure polyoses content, color
For white.
Coriolan for the checking present invention has stronger immunologic enhancement, now carries out following zoopery.
1st, anti-Lewis lung cancer activity is tested
Cleaning grade C57BL/6 mouse 30,18~20g of mouse weight are taken, are randomly divided into 3 groups, every group 10, be respectively real
Test group, negative control group and positive controls.
Take eugonic Mice Bearing Lewis Lung Cancer tumour cell that homogenate about 1~2 × 10 is made in conventional manner7cfu/ml
Cancer cell suspension, after the right armpit subcutaneous vaccination 0.2ml cancer cell suspensions of every mouse, 24h, each group starts daily mouth
Clothes are administered once, and the coriolan of embodiment 1, negative control group administration physiological saline, positive controls administration is administered in experimental group
The coriolan of reference examples 1, dosage see the table below 1, successive administration 10 days.Next day puts to death all animals, interception foot after drug withdrawal
Toe is weighed and calculates tumor control rate.Inhibition rate of tumor growth is calculated by following equation, inhibiting rate (%)=(negative control group knurl
Weight-experimental group/positive controls knurl weight)/negative control group knurl weight × 100.
Table 1
| Dosage (mg/kg) | Animal increases weight (g) | Knurl weight (g) | Inhibiting rate (%) | |
| Negative control group | 1ml | 3.0 | 2.80±0.33 | |
| Positive controls | 2000 | 2.9 | 1.79±0.07 | 36.07* |
| Experimental group | 2000 | 1.8 | 1.06±0.16 | 62.14** |
Note：Compared with negative control group, * p ＜ 0.05, * * p ＜ 0.01.
As seen from the above table, the coriolan extracted using conventional method is reached to the inhibiting rate of Mice Bearing Lewis Lung Cancer
36.07%, and the coriolan that the present invention extracts is up to 62.14% to the inhibiting rate of Mice Bearing Lewis Lung Cancer, hence it is evident that better than routine
Coriolan.
2nd, anti-C-26 colon cancer reactives experiment
Cleaning grade C57BL/6 mouse 30,18~20g of mouse weight are taken, are randomly divided into 3 groups, every group 10, be respectively real
Test group, negative control group and positive controls.
Take eugonic mouse C-26 colon cancer tumours cell that homogenate about 1~2 × 10 is made in conventional manner7cfu/ml
Cancer cell suspension, after the right armpit subcutaneous vaccination 0.2ml cancer cell suspensions of every mouse, 24h, each group starts daily mouth
Clothes are administered once, and the coriolan of embodiment 1, negative control group administration physiological saline, positive controls administration is administered in experimental group
The coriolan of reference examples 1, dosage see the table below 2, successive administration 10 days.Next day puts to death all animals, interception foot after drug withdrawal
Toe is weighed and calculates tumor control rate.Inhibition rate of tumor growth is calculated by following equation, inhibiting rate (%)=(negative control group knurl
Weight-experimental group/positive controls knurl weight)/negative control group knurl weight × 100.
Table 2
| Dosage (mg/kg) | Animal increases weight (g) | Knurl weight (g) | Inhibiting rate (%) | |
| Negative control group | 1ml | 3.8 | 2.99±0.26 | |
| Positive controls | 2000 | 3.8 | 1.92±0.23 | 35.79** |
| Experimental group | 2000 | 1.2 | 1.19±0.06 | 60.20** |
Note：Compared with negative control group, * p ＜ 0.05, * * p ＜ 0.01.
As seen from the above table, the coriolan extracted using conventional method is reached to the inhibiting rate of mouse C-26 colon cancers
35.79%, and the coriolan that the present invention extracts is up to 60.20% to the inhibiting rate of Mice Bearing Lewis Lung Cancer, hence it is evident that better than routine
Coriolan.
Claims (2)
1. the extracting method of coriolan, it is characterised in that：Comprise the following steps：
1) rainbow conk is crushed, extracted 2~3 times with 6~12 times of 80~95v% of weight alcohol refluxs, flow back 2~3h every time, filtering,
Take filter residue standby；
2) the water refluxing extraction 2~3 times by filter residue with 6~12 times of weight, 2~3h is extracted every time, filtering, it is standby to collect filtrate；
3) medicinal extract that relative density at 60 DEG C is 1.1~1.3 is concentrated the filtrate to, adds ethanol precipitation, centrifuging to precipitate；
4) precipitation is washed respectively with the absolute ethyl alcohol of 2~4 times of weight, acetone successively, the precipitation after washing is steamed with 2~4 times
Distilled water dissolves, and crosses the hollow-fibre membrane that molecular cut off is 1,000,000, collects concentrate, concentrate then is crossed into molecular cut off
For 1,500,000 hollow-fibre membrane, permeate centrifugation is collected, filtering, takes filtrate standby；
5) filtrate is taken to be spray-dried, as coriolan.
2. the extracting method of coriolan according to claim 1, it is characterised in that：In step 3), added into medicinal extract
90~95v% ethanol to ethanol content is 80~85v%.
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| CN109106731B (en) * | 2018-11-13 | 2021-07-20 | 广东省微生物研究所（广东省微生物分析检测中心） | Preparation method of coriolus versicolor extract and application of coriolus versicolor extract in reducing uric acid |
| CN114805626B (en) * | 2022-05-10 | 2023-02-03 | 湖南中药谷集团研究院有限公司 | Polysaccharide with anticancer activity, its preparation method and application in preparing anticancer drugs |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651076A (en) * | 2004-12-29 | 2005-08-10 | 江苏华源药业有限公司 | Method of extracting rainbow conk saccharide peptide for injection in rainbow conk saccharided peptide crude product obtained from submerged fermentation |
| CN102911279A (en) * | 2012-10-23 | 2013-02-06 | 江苏神华药业有限公司 | Polysaccharide-peptide and liposome thereof |
| CN104231100A (en) * | 2014-09-04 | 2014-12-24 | 中山安荞生物科技有限公司 | Method for extracting coriolus versicolor polysaccharides |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651076A (en) * | 2004-12-29 | 2005-08-10 | 江苏华源药业有限公司 | Method of extracting rainbow conk saccharide peptide for injection in rainbow conk saccharided peptide crude product obtained from submerged fermentation |
| CN102911279A (en) * | 2012-10-23 | 2013-02-06 | 江苏神华药业有限公司 | Polysaccharide-peptide and liposome thereof |
| CN104231100A (en) * | 2014-09-04 | 2014-12-24 | 中山安荞生物科技有限公司 | Method for extracting coriolus versicolor polysaccharides |
Non-Patent Citations (1)
| Title |
|---|
| 云芝多糖分离、纯化、药理活性及其作用机制研究;孙晓波;《中国优秀博硕士学位论文全文数据库（博士） 医药卫生科技辑》;20041215(第4期);E057-15 * |
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Effective date of registration: 20200426 Address after: No. 181, erbatou, taimuyang village, Qinyu Town, Fuding City, Ningde City, Fujian Province Patentee after: Deng Weixiao Address before: Yingbin Road, Xingan County Guilin 541300 the Guangxi Zhuang Autonomous Region (Guilin Lingqu chemical factory warehouse) Patentee before: GUILIN APLANAT BOTANICAL RESOURCES Co.,Ltd. |
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Effective date of registration: 20200512 Address after: 057750 No.2 Chaoyang Road, Guantao Economic Development Zone, Handan City, Hebei Province Patentee after: Hebei Yiwei daily necessities Co., Ltd Address before: No. 181, erbatou, taimuyang village, Qinyu Town, Fuding City, Ningde City, Fujian Province Patentee before: Deng Weixiao |
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