CN100480276C - Process of preparing high purity astragalus polysaccharide - Google Patents
Process of preparing high purity astragalus polysaccharide Download PDFInfo
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- CN100480276C CN100480276C CNB200710013890XA CN200710013890A CN100480276C CN 100480276 C CN100480276 C CN 100480276C CN B200710013890X A CNB200710013890X A CN B200710013890XA CN 200710013890 A CN200710013890 A CN 200710013890A CN 100480276 C CN100480276 C CN 100480276C
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Abstract
The process of preparing high purity astragalus polysaccharide includes the following steps: crushing astragalus root and constant temperature microwave extracting in distilled water at 50-75 deg.c, centrifuging; taking supernatant and decolorizing with active carbon to obtain solution of astragalus polysaccharide; concentrating and adding alcohol via stirring; centrifuging and dissolving the precipitate in distilled water; filtering the gel and chromatography; detecting the collected liquid at 280 nm wavelength and collecting eluted liquid at the absorption peak; and vacuum freeze drying to obtain the astragalus polysaccharide product. The present invention has astragalus polysaccharide product purity over 90 %, relaxed preparing process, low cost and other advantages.
Description
Technical field
The present invention relates to a kind of preparation method of vegetable polysaccharides, relate in particular to a kind of method for preparing high purity astragalus polysaccharide of from Chinese medicine astragalus, separating.
Background technology
The Radix Astragali (Radix Astragali) is called the continuous Radix Astragali, and English MilkvetchRoot is the root of leguminous plants Radix Astagali Astragalus membranaceus Bge.var.mongholicus (Bge.) Hsiao.The Radix Astragali has invigorating QI to consolidate the body surface resistance, expelling pus and toxin by strengthening QI, diuresis, the effect of expelling pus and promoting granulation; Clinical be usually used in the deficiency of vital energy weak, rush down illnesss such as prolapse of the anus, spontaneous perspiration, oedema, sub-official's prolapsus, chronic nephritis proteinuria, diabetes, open sore part disunion of a specified duration for a long time.
In recent decades, it is found that the polysaccharide that extracts has extremely important and special physiologically active from plant.These polysaccharide have participated in the comings and goings of cell in the life science, have diversified biological function, and wherein astragalus polysaccharides is exactly study more deeply in these polysaccharide a kind of.
Astragalus polysaccharides is one of effective constituent of Chinese medicine astragalus, is separated obtaining by Chinese medicine astragalus, and modern scientific research confirms: astragalus polysaccharides has raise immunity, protection cardiovascular systems, two-ways regulation blood sugar and multiple pharmacological effect such as hypotensive.Further experimental data shows, astragalus polysaccharides can stimulate splenocyte propagation, improves LAK cell and NK cytoactive, corrects the immunologic hypofunction of caused by cyclophosphamide; Other has experiment confirm, and it can improve the body substance metabolism, corrects the Acute Myocardial Ischemia in Rats that posterior pituitary causes, the ST section is raise, and the T ripple is inverted and is alleviated etc.The prompting astragalus polysaccharides has pharmaceutical value, can be widely used in that to correct cancer patients's chemotherapeutic immunity power low, diseases such as control diabetic subject's cardiovascular pathological changes and treatment gastritis.This class number of patients is huge day by day in China, but the effectively preventing means are few at present, and therefore, exploitation of astragalus polysaccharides preparation and products thereof and application are to clinical very urgent.
Though relating to the report of astragalus polysaccharides extraction and separation method also has, but the astragalus polysaccharides product that it obtains is comparatively rough, purity is not high, can't satisfy the formulation preparation requirement that improves day by day at present, Given this, found a kind of task of top priority that the method for preparing high purity astragalus polysaccharide should be astragalus polysaccharides research that from Chinese medicine astragalus, separates.
Summary of the invention
At the deficiencies in the prior art, the problem to be solved in the present invention provides a kind of method for preparing high purity astragalus polysaccharide of separating from Chinese medicine astragalus.
The method for preparing high purity astragalus polysaccharide of the present invention, form by following step:
(1) astragalus membranaceus powder being broken to particle diameter is 200～300 μ m, with water by quality ratio, the amount by 1: 6～10 is dipped in the distilled water, 50～75 ℃ are carried out the constant temperature microwave extraction, extraction conditions: microwave power 250W, 11～15 minutes time, repeat 2～3 times;
(2) with 3000～5000rpm centrifugal (1) described extraction mixed solution 12～16min, get supernatant liquor,, get astragalus polysaccharides aqueous extract solution with activated carbon decolorizing;
(3) concentrating astragalus polysaccharides aqueous extract solution to proportion is 1.15～1.25;
(4) to concentrate the amount of 6～8 times of back astragalus polysaccharides aqueous extract liquor capacities, in astragalus polysaccharides aqueous extract solution, add 70%～95% ethanol, stir 20～36min;
(5) with the described liquid 12～16min that mixes of 3000～5000rpm centrifugal (4), get precipitation and with distill water-soluble rare be 1.05～1.15 solution to proportion;
(6) choosing internal diameter and aspect ratio is 1: 30～50 chromatography column; Add dextrane gel in post, add-on is 65%～85% of a column volume;
(7) solution of gel permeation chromatography step (5), elutriant is a distilled water, flow velocity is 0.3～0.6ml/min, is that 280nm detects collection liquid with the wavelength, selects the absorption peak elutriant to collect, and promptly gets extraction of Astragalus Polysaccharides in Astragalus liquid;
(8) conventional vacuum lyophilization extraction of Astragalus Polysaccharides in Astragalus liquid gets high purity astragalus polysaccharide.
In the above-mentioned method for preparing high purity astragalus polysaccharide, preferred 60～65 ℃ of the described temperature of step (1).
In the above-mentioned method for preparing high purity astragalus polysaccharide, step (2) or the preferred 4000rpm of (5) described centrifugal speed, the preferred 15min of centrifugation time.
In the above-mentioned method for preparing high purity astragalus polysaccharide, the internal diameter of the described chromatography column of step (6) and aspect ratio preferred 1: 35～45.
In the above-mentioned method for preparing high purity astragalus polysaccharide, the described dextrane gel amount with molecular sieve effect that adds in post of step (6) is preferably 70%～80% of column volume.
In the above-mentioned method for preparing high purity astragalus polysaccharide, the described dextrane gel of step (6) is selected SephadexG-150 or SephadexG-200 for use.
Wherein, the described dextrane gel of step (6) SephadexG-150 most preferably.
In the above-mentioned method for preparing high purity astragalus polysaccharide, the preferred 0.4ml/min of the described flow velocity of step (7).
The method for preparing high purity astragalus polysaccharide of utilizing the present invention to propose, can prepare purity and reach astragalus polysaccharides more than 93%, solve astragalus polysaccharides at all and separated difficulty, purity is low, the unsettled deficiency of quality, and method equipment of the present invention and environmental requirement are low, method is easy, cost is low, and is practical flexibly, is beneficial to popularization, development and application.
The astragalus polysaccharides that adopts separation purification method of the present invention to prepare will have very wide prospect in clinical application and protective foods exploitation.
Embodiment
Embodiment 1:
(1) astragalus membranaceus powder being broken to particle diameter is 200～300 μ m, with water by quality ratio, be dipped in the distilled water by 1: 8 amount, 65 ℃ are carried out the constant temperature microwave extraction, extraction conditions: microwave power 250W, 13 minutes time, repeat 3 times;
(2) with 4000rpm centrifugal (1) described extraction mixed solution 15min, get supernatant liquor,, get astragalus polysaccharides aqueous extract solution with activated carbon decolorizing;
(3) concentrating astragalus polysaccharides aqueous extract solution to proportion is 1.15～1.20;
(4) to concentrate the amount of 7 times of back astragalus polysaccharides aqueous extract liquor capacities, in astragalus polysaccharides aqueous extract solution, add 85% ethanol, stir 26min;
(5) with the described liquid 15min that mixes of 4000rpm centrifugal (4), get precipitation and with distill water-soluble rare be 1.05～1.10 solution to proportion;
(6) choosing internal diameter and aspect ratio is 1: 40 chromatography column; Add the SephadexG-150 dextrane gel in post, add-on is 75% of a column volume;
(7) solution of gel permeation chromatography step (5), elutriant is a distilled water, flow velocity is 0.4ml/min, is that 280nm detects collection liquid with the wavelength, selects the absorption peak elutriant to collect, and promptly gets extraction of Astragalus Polysaccharides in Astragalus liquid;
(8) conventional vacuum lyophilization extraction of Astragalus Polysaccharides in Astragalus liquid gets high purity astragalus polysaccharide.
Embodiment 2:
(1) astragalus membranaceus powder being broken to particle diameter is 200～300 μ m, with water by quality ratio, be dipped in the distilled water by 1: 6 amount, 75 ℃ are carried out the constant temperature microwave extraction, extraction conditions: microwave power 250W, 11 minutes time, repeat 2 times;
(2) with 3000rpm centrifugal (1) described extraction mixed solution 16min, get supernatant liquor,, get astragalus polysaccharides aqueous extract solution with activated carbon decolorizing;
(3) concentrating astragalus polysaccharides aqueous extract solution to proportion is 1.15～1.2;
(4) to concentrate the amount of 6 times of back astragalus polysaccharides aqueous extract liquor capacities, in astragalus polysaccharides aqueous extract solution, add 70% ethanol, stir 20min;
(5) with the described liquid 16min that mixes of 3000rpm centrifugal (4), get precipitation and with distill water-soluble rare be 1.05～1.1 solution to proportion;
(6) choosing internal diameter and aspect ratio is 1: 30 chromatography column; Add the SephadexG-200 dextrane gel in post, add-on is 65% of a column volume;
(7) solution of gel permeation chromatography step (5), elutriant is a distilled water, flow velocity is 0.6ml/min, is that 280nm detects collection liquid with the wavelength, selects the absorption peak elutriant to collect, and promptly gets extraction of Astragalus Polysaccharides in Astragalus liquid;
(8) conventional vacuum lyophilization extraction of Astragalus Polysaccharides in Astragalus liquid gets high purity astragalus polysaccharide.
Embodiment 3:
(1) astragalus membranaceus powder being broken to particle diameter is 200～300 μ m, with water by quality ratio, be dipped in the distilled water by 1: 10 amount, 50 ℃ are carried out the constant temperature microwave extraction, extraction conditions: microwave power 250W, 15 minutes time, repeat 3 times;
(2) with 5000rpm centrifugal (1) described extraction mixed solution 12min, get supernatant liquor,, get astragalus polysaccharides aqueous extract solution with activated carbon decolorizing;
(3) concentrating astragalus polysaccharides aqueous extract solution to proportion is 1.2～1.25;
(4) to concentrate the amount of 6～8 times of back astragalus polysaccharides aqueous extract liquor capacities, in astragalus polysaccharides aqueous extract solution, add 95% ethanol, stir 36min;
(5) with the described liquid 12min that mixes of 5000rpm centrifugal (4), get precipitation and with distill water-soluble rare be 1.1～1.15 solution to proportion;
(6) choosing internal diameter and aspect ratio is 1: 50 chromatography column; Add the SephadexG-150 dextrane gel in post, add-on is 85% of a column volume;
(7) solution of gel permeation chromatography step (5), elutriant is a distilled water, flow velocity is 0.3ml/min, is that 280nm detects collection liquid with the wavelength, selects the absorption peak elutriant to collect, and promptly gets extraction of Astragalus Polysaccharides in Astragalus liquid;
(8) conventional vacuum lyophilization extraction of Astragalus Polysaccharides in Astragalus liquid gets high purity astragalus polysaccharide.
The Microwave Extraction Equipment that relates among the present invention is WTS03-1 type (the outstanding full microwave equipment company limited in Nanjing), perhaps MAS-I type (the normal pressure microwave-assisted synthesizes/the extractive reaction instrument, Xinyi Microwave Chemistry Tech Co., Ltd.).
" SephadexG-150, the SephadexG-200 " that relates among the present invention is pharmacia company product.The company of China sale by agent pharmacia product has: " Beijing Baeyer enlightening biotech company, the clear wooden Room 910, No. 1 building, magnificent garden (100080) of No. 4 water in North Second Street, Zhongguancun, Haidian District, Beijing City; Phone (TEL): 010-82642396; Fax (FAX): 010-82642395; Or Beijing nation decides Tyke biotech company, the address: No. 48, Fuxing Road, Haidian District, Beijing City, postcode: 100039; Phone: 86-010-68287358; Fax: 010-68287361.”
Claims (8)
1. method for preparing high purity astragalus polysaccharide, form by following step:
(1) astragalus membranaceus powder being broken to particle diameter is 200～300 μ m, with water by quality ratio, be dipped in the distilled water by the amount of 1:6～10,50～75 ℃ are carried out the constant temperature microwave extraction, extraction conditions: microwave power 250W, 11～15 minutes time, repeat 2～3 times;
(2) with 3000～5000rpm centrifugal (1) described extraction mixed solution 12～16min, get supernatant liquor,, get astragalus polysaccharides aqueous extract solution with activated carbon decolorizing;
(3) concentrating astragalus polysaccharides aqueous extract solution to proportion is 1.15～1.25;
(4) to concentrate the amount of 6～8 times of back astragalus polysaccharides aqueous extract liquor capacities, in astragalus polysaccharides aqueous extract solution, add 70%～95% ethanol, stir 20～36min;
(5) with the described liquid 12～16min that mixes of 3000～5000rpm centrifugal (4), get precipitation and with distill water-soluble rare be 1.05～1.15 solution to proportion;
(6) choose the chromatography column that internal diameter and aspect ratio are 1:30～50; Add dextrane gel in post, add-on is 65%～85% of a column volume;
(7) solution of gel permeation chromatography step (5), elutriant is a distilled water, flow velocity is 0.3～0.6ml/min, is that 280nm detects collection liquid with the wavelength, selects the absorption peak elutriant to collect, and promptly gets extraction of Astragalus Polysaccharides in Astragalus liquid;
(8) conventional vacuum lyophilization extraction of Astragalus Polysaccharides in Astragalus liquid gets high purity astragalus polysaccharide.
2. prepare the method for high purity astragalus polysaccharide according to claim 1, it is characterized in that, the described temperature of step (1) is 60～65 ℃.
3. prepare the method for high purity astragalus polysaccharide according to claim 1, it is characterized in that, step (2) or (5) described centrifugal speed are 4000rpm, and centrifugation time is 15min.
4. prepare the method for high purity astragalus polysaccharide according to claim 1, it is characterized in that, the internal diameter and the aspect ratio of the described chromatography column of step (6) are 1:35～45.
5. prepare the method for high purity astragalus polysaccharide according to claim 1, it is characterized in that, the described dextrane gel amount with molecular sieve effect that adds in post of step (6) is 70%～80% of a column volume.
6. prepare the method for high purity astragalus polysaccharide according to claim 1, it is characterized in that, the described dextrane gel of step (6) is selected SephadexG-150 or SephadexG-200 for use.
7. as preparing the method for high purity astragalus polysaccharide as described in the claim 6, it is characterized in that the described dextrane gel of step (6) is selected SephadexG-150 for use.
8. prepare the method for high purity astragalus polysaccharide according to claim 1, it is characterized in that, the described flow velocity of step (7) is 0.4ml/min.
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| CNB200710013890XA CN100480276C (en) | 2007-03-29 | 2007-03-29 | Process of preparing high purity astragalus polysaccharide |
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| CN100480276C true CN100480276C (en) | 2009-04-22 |
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102949462B (en) * | 2011-08-30 | 2014-09-17 | 河南金尔康动物药业有限公司 | Veterinary Shuanghuanglian preparation and preparation method for same |
| CN102993321B (en) * | 2012-09-27 | 2015-06-03 | 河南牧翔动物药业有限公司 | Preparation method of astragalus polysaccharide |
| CN103073650B (en) * | 2012-11-26 | 2015-01-07 | 兰州大学 | Red stilbene polysaccharide 4 and effective component preparation and application |
| CN103524632B (en) * | 2013-03-21 | 2016-06-08 | 王喜军 | The quick reparation technology that a kind of astragalus polysaccharides produces |
| CN104208662A (en) * | 2013-09-30 | 2014-12-17 | 郑州后羿制药有限公司 | Compound injection for treating poultry viral diseases and preparation method thereof |
| CN105440147A (en) * | 2015-12-14 | 2016-03-30 | 陈燕妮 | Process for producing astragalus polysaccharide by using microwave technology |
| KR102206714B1 (en) * | 2016-11-10 | 2021-01-25 | 피토헬스 코포레이션 | Method for enhancing effect of immunotherapy for cancer |
| CN107334808B (en) * | 2017-08-23 | 2020-09-04 | 齐齐哈尔医学院 | Astragalus extract preparation with antibacterial activity |
| CN108084288A (en) * | 2017-12-26 | 2018-05-29 | 浦江县美泽生物科技有限公司 | efficient astragalus polyose extracting method |
| CN109091708A (en) * | 2018-08-15 | 2018-12-28 | 杭州市萧山区中医院 | Collagen-fibroin-chitosan/astragalus polyose engineering skin three-dimensional rack |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1854158A (en) * | 2005-04-25 | 2006-11-01 | 蒋来高 | Extraction of astragalus mongholicus polysaccharose by super-critical fluid technology |
| CN1880342A (en) * | 2005-06-17 | 2006-12-20 | 上海医药工业研究院 | Process for extracting polysaccharide from astragalus root using microwave |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1854158A (en) * | 2005-04-25 | 2006-11-01 | 蒋来高 | Extraction of astragalus mongholicus polysaccharose by super-critical fluid technology |
| CN1880342A (en) * | 2005-06-17 | 2006-12-20 | 上海医药工业研究院 | Process for extracting polysaccharide from astragalus root using microwave |
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