CN1199651C - Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease - Google Patents
Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease Download PDFInfo
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- CN1199651C CN1199651C CNB031133304A CN03113330A CN1199651C CN 1199651 C CN1199651 C CN 1199651C CN B031133304 A CNB031133304 A CN B031133304A CN 03113330 A CN03113330 A CN 03113330A CN 1199651 C CN1199651 C CN 1199651C
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- radix astragali
- total saponins
- ethanol
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Abstract
The present invention relates to astragalus root total saponin having the function of protecting myocardial damage. The preparation method of the astragalus root saponin comprises the following steps: dissolving the extract of astragalus root alcohol with water, adsorbing the extract of astragalus root alcohol with a macroreticular resin column, respectively eluting the extract of astragalus root alcohol with water and alcohol containing water, combining 70% to 98% of alcohol eluent, and drying the alcohol eluent to obtain extractum; dissolving the extractum with methanol and adding diatomite to stir, with the proportion of the diatomite to the extractumof 3 to 1:1; carrying out hot reflux extraction with anhydrous alcohol and evaporating the extracted liquid to obtain a saponin extractive. The content of total saponin reaches 85% to 90%, and the transferring rate of the total saponin is from 80% to 85%.
Description
Technical field
The present invention obtains to have the highly purified Radix Astragali total saponins of protection myocardial damage effect by the improvement of preparation method.
Background technology
The Radix Astragali (Radix astragali) is the dry root of leguminous plant Radix Astagali Astragalus membranaceus (Fisch.) Bge.var mongholicus (Bge.) Hsiao or Radix Astragali Astragalus membranaceus (Fisch.) Bge., property is sweet, warm, return lung, spleen channel, invigorating QI to consolidate the body surface resistance, the diuresis poison holding, evacuation of pus, expelling pus and promoting granulation.Modern chemistry studies have shown that compositions such as containing Saponin, polysaccharide, flavone, aminoacid, phenoloid and trace element in the Milkvetch Root.Astragaloside tool heart tonifying function can be used for treating heart failure, and the recovery of cardiac function behind the acute myocardial infarction and the removing tool of oxygen-derived free radicals are obviously acted on.Especially in recent years the Radix Astragali is used for the treatment of the research of viral myocarditis; find that the Radix Astragali can suppress duplicating of Coxsackie virus; viral infection myocardial cell abnormal electrical activity there is reverse effect; through comparative study the protective effect of Radix Astragali heterogeneity to the viral infection myocardial cell injury, confirmed that the main effective ingredient of the Radix Astragali is a Radix Astragali total saponins.But astragaloside content is lower, and root contains a large amount of glucides, and extraction separation is difficulty relatively.
Existing extraction and separation process has: water extraction, alcohol extracting method, resin adsorption method, n-butanol extraction and supercritical CO
2Method.For removing sucrose, the monosaccharide in the medical material, and useful cold-maceration (referring to: Sugarless active constituent-enriched extractive and goods thereof, CN1171950A); Decoction and alcohol sedimentation technique (referring to: a kind of process for refining astragalus injection, CN1273116A).
Summary of the invention
The technical problem to be solved in the present invention is by improving the extraction separation method of existing Radix Astragali effective site astragaloside, obtaining total Saponin that content height, purity improve, guaranteeing the stable of curative effect, and can have clearer and more definite quality control index.
For addressing the above problem, the invention provides following technical solution:
A kind of astragaloside is characterized in that available following method obtains:
1) get the ethanol extraction extractum of the Radix Astragali, use water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, the Radix Astragali total saponins crude product;
2) or, get the ethanol extraction extractum of the Radix Astragali, use water dissolution, earlier with the saturated n-butanol extraction of NaOH of 0.1N, concentrated extract is to the extractum shape, the reuse water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution, merging 70% and 70%-98% ethanol elution get the Radix Astragali total saponins crude product;
3) or, get the ethanol extraction extractum of the Radix Astragali, use dissolve with methanol, add kieselguhr and mix thoroughly, extract with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness, the Radix Astragali total saponins crude product.
Described astragaloside is characterized in that:
With 1) in obtain the Radix Astragali total saponins crude product, use water dissolution, go up macroporous resin column once more, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, Radix Astragali total saponins;
Perhaps, with 1) in obtain the Radix Astragali total saponins crude product, use dissolve with methanol, add kieselguhr and mix thoroughly, extract with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness, Radix Astragali total saponins;
Perhaps, with 2) in obtain the Radix Astragali total saponins crude product, use dissolve with methanol, add kieselguhr and mix thoroughly, extract with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness, Radix Astragali total saponins;
Perhaps, with 3) in obtain the Radix Astragali total saponins crude product, use water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, Radix Astragali total saponins;
Perhaps, with 3) in obtain the Radix Astragali total saponins crude product, use water dissolution, earlier with the saturated n-butanol extraction of alkaline solution, as the saturated n-butanol extraction of NaOH with 0.1N, concentrated extract is to the extractum shape, the reuse water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution, merge the 70%-98% ethanol elution, get Radix Astragali total saponins.
The model of big pore resin column of the present invention is D101, AB-8; Ethanol extraction extractum and diatomaceous weight ratio are 1: 1-3; Ethanol extraction extractum and diatomaceous weight ratio are for being 1: 2 preferably.
Be used to prevent and treat the pharmaceutical composition of cardiovascular disease, wherein contain in the claim 1 or 2 for the treatment of effective dose each astragaloside and pharmaceutically acceptable carrier.
The present invention removes glucides a large amount of in the Radix Astragali by selected macroporous resin column chromatography and certain elution requirement, has significantly improved the content of total Saponin.If macroporous resin column on twice can extremely significantly improve the total saponin content of gained extractum.The common impurity that influence the quality of the pharmaceutical preparations such as pigment for Chinese medicine, general activated carbon commonly used is removed, but showing, experimental data of the present invention removes the yield that impurity such as pigment often greatly reduce astragaloside with activated carbon, and the present invention takes kieselguhr to mix thoroughly, with dehydrated alcohol hot reflux absorption method, the effect of removing impurity such as pigment is very obvious, and the yield of astragaloside is also high.The extracting mode of total Saponin, extraction solvent, solubilization dosage, return time, extraction time are to form through orthogonal Design Research in the technology of the present invention, have guaranteed resulting Radix Astragali total saponins content height, and the rate of transform is greater than 80%.The present invention compared with prior art has the total saponin content height, cost is low, organic solvent-free is residual and advantage such as production process environmentally safe.
Astragaloside of the present invention has the effect of significant protection myocardial damage.For medicine, the purity and the curative effect of its treatment component are closely related.The present invention is by improving the extraction separation method of existing Radix Astragali effective site astragaloside, obtain total saponin content greater than 80% astragaloside, the raising of purity, guaranteed in the stable curative effect of preventing and treating aspect the cardiovascular disease, astragaloside and contain the steady quality of the pharmaceutical composition of astragaloside, quality control index is clear and definite.
The optimised process of research Radix Astragali total saponins (content is greater than 80%), this prepared product have the effect of protection myocardial damage.
Its research step and detailed data are as follows:
1. medical material
Select the dry root of Radix Astagali Astragalus membranaceus (Fisch.) Bge.varmongholicus (Bge.) Hsiao or Radix Astragali Astragalus membranaceus (Fisch.) Bge. for use.
2. the influence of total Saponin extracting mode:
Get and be ground into 40 purpose Milkvetch Root powder 50g, the accurate title, decide, and extracts by following method respectively:
(1) water extract-alcohol precipitation: the material 50g that gets it filled, adding distil water 200ml, heating extraction 3 times, each 1.5h, water liquid is centrifugal, supernatant merges, and is concentrated in right amount, adds 95% ethanol precipitation, leaves standstill 12h, and is centrifugal, gets supernatant and merges, concentrate extractum.
(2) 50% alcohol extracts: the material 50g that gets it filled, add 50% ethanol 200ml, 90 ℃ of reflux 3 times, each 1.5h, merge extractive liquid,, concentrate extractum.
(3) 70% alcohol extracts: the material 50g that gets it filled, add 70% ethanol 200ml, 90 ℃ of reflux 3 times, each 1.5h, merge extractive liquid,, concentrate extractum.
(4) 95% ethanol extracts: the material 50g that gets it filled, add 95% ethanol 200ml, 90 ℃ of reflux 3 times, each 1.5h, merge extractive liquid,, concentrate extractum.
Experimental results show that: yield of extract is 50% alcohol extract＞70% alcohol extract＞95% alcohol extract＞water extract-alcohol precipitation.
3, the separation of Radix Astragali total saponins, purification, concentrated and drying process
3.1 isolation and purification technology
Get Radix Astagali or Radix Astragali medical material,, extract 3 times, add 10 times of amounts of 50-95% ethanol at every turn, extract 1.5h/ time with the 50-95% alcohol heating reflux earlier with pulverizing medicinal materials.Merge extractive liquid, gets extractum A.
With an amount of dissolved in distilled water, last macroporous resin column respectively with distilled water, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 98% ethanol elution, merges the 70%-98% ethanol elution with extractum A, and evaporate to dryness gets extractum B.
With extractum B with dissolve with methanol, adding kieselguhr mixes thoroughly, kieselguhr and extractum ratio are 3-1: 1, with 2: 1 for well, extract 3 times gained extracting solution evaporate to dryness with the dehydrated alcohol hot reflux, get extract I, calculate with astragaloside, this extract contains total Saponin and reaches 85%-90%, and total Saponin rate of transform is at 80%-85%.
If with this extract I with dissolved in distilled water, last macroporous resin column, respectively with distilled water, 20%, 30%, 40%, 50%, 60%, 70%, 80% ethanol elution, merge 50%-80% ethanol stream part, evaporate to dryness gets extract II, calculates with astragaloside, this extract contains total Saponin and reaches more than 98%, and total Saponin rate of transform is at 75%-80%.
3.2 concentrate and drying process
With 70%-98% ethanol elution decompression and solvent recovery, vacuum drying promptly gets total Saponin position.
Radix Astragali total saponins of the present invention is to cultivating the protective effect of myocardial cell
1, Radix Astragali total saponins is to cultivating the influence of myocardial cell calcium overload
(1) Radix Astragali total saponins is to the influence of myocardial cell free calcium
Radix Astragali total saponins is intervened cell separately to free calcium ions ([Ca in the born of the same parents
2+]
i) there is not influence, but do the time spent simultaneously with isoproterenol, Radix Astragali total saponins can suppress [the Ca that isoproterenol causes
2+]
iIncrease.With isoproterenol relatively, Radix Astragali total saponins from the low concentration to the high concentration (10,100,200mgml
-1), reduce [Ca respectively
2+]
i39%, 40%, 51% (P＜0.05).
(2) Radix Astragali total saponins is to myocardial cell sarcoplasmic reticulum calcium effects of load
Radix Astragali total saponins is intervened cell does not separately have influence to sarcoplasmic reticulum calcium load (Sarcoplasmic reticulum Calcium load), isoproterenol can make sarcoplasmic reticulum calcium load increase by 145% (P＜0.01), do the time spent simultaneously with isoproterenol, Radix Astragali total saponins can suppress the increase of the sarcoplasmic reticulum calcium load that isoproterenol causes, from the low concentration to the high concentration, reduce by 11% (P＞0.05) respectively, 35% (P＜0.05), 51% (P＜0.05).
(3) Radix Astragali total saponins is to myocardial cell sarcoplasmic reticulum Ca
2+The active influence of-ATPase
Radix Astragali total saponins is intervened cell separately can make sarcoplasmic reticulum Ca
2+-ATPase is active to be increased, but does the time spent simultaneously with isoproterenol, and Radix Astragali total saponins can suppress the sarcoplasmic reticulum Ca that isoproterenol causes
2+The active increase of-ATPase can suppress 34% (P＜0.05) in the high concentration group.
2, Radix Astragali total saponins is to cultivating the detection of myocardial cell injury index (Troponin I)
Radix Astragali total saponins acts on the cultivation myocardial cell, detects myocardial cell damage criterion---Troponin I (cTnI) in the supernatant, finds to compare the cTnI no change with the normal control group, shows that Radix Astragali total saponins is to the myocardial cell not damaged.
The specific embodiment
Select in following 2 kinds of Milkvetch Roots that the Pharmacopoeia of the People's Republic of China (version in 2000) records wherein a kind of for use, that is: the dry root of Radix Astagali Astragalus membranaceus (Fisch.) Bge.var mongholicus (Bge.) Hsiao or Radix Astragali Astragalus membranaceus (Fisch.) Bge..The properties and characteristics of medical material, physicochemical identification feature meet the Pharmacopoeia of the People's Republic of China (version was an one in 2000) " Radix Astragali " item content down.
Embodiment 1
Take by weighing Milkvetch Root 20kg, pulverize, cross 20 mesh sieves, add 50% ethanol of 8 times of amounts, 80 ℃ are extracted 3 times, each .5 hour.Merge extractive liquid, gets extractum A.With extractum A with an amount of dissolved in distilled water, with the saturated n-butanol extraction of 0.1N NaOH 3 times, combining extraction liquid, get extractum B, with extractum B with dissolved in distilled water, last macroporous resin column, respectively with distilled water,, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 98% ethanol elution, merge the 70%-98% ethanol elution, evaporate to dryness gets extract I, calculates with astragaloside, this extract contains total Saponin and reaches 75.6%, and total Saponin rate of transform is 80.0%.
Embodiment 2
Take by weighing Milkvetch Root 20kg, pulverize, cross 20 mesh sieves, add 95% ethanol of 10 times of amounts, 90 ℃ are extracted each 1.5 hours 3 times.Merge extractive liquid, gets extractum A.With extractum A with an amount of dissolved in distilled water, with the saturated n-butanol extraction of 0.1N NaOH 3 times, combining extraction liquid, get extractum B, with dissolved in distilled water, last macroporous resin column is respectively with distilled water, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 98% ethanol elution with extractum B, merge the 70%-98% ethanol elution, evaporate to dryness gets extractum C.Extractum C with dissolve with methanol, is added kieselguhr and mixes thoroughly, and kieselguhr and extractum ratio are 2: 1, extract 3 times with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness gets extract I, calculate with astragaloside, this extract contains total Saponin and reaches 85.2%, and total Saponin rate of transform is 72.5%.
Embodiment 3
Take by weighing Milkvetch Root 20kg, pulverize, cross 20 mesh sieves, add 70% ethanol of 10 times of amounts, 80 ℃ are extracted each 1.5 hours 3 times.Merge extractive liquid, gets extractum.With an amount of dissolved in distilled water, last macroporous resin column respectively with distilled water, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 98% ethanol elution, merges the 70%-98% ethanol elution with this extractum, and evaporate to dryness gets extractum B.Extractum B with dissolve with methanol, is added kieselguhr and mixes thoroughly, and kieselguhr and extractum ratio are 2: 1, extract 3 times with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness gets extract I, calculate with astragaloside, this extract contains total Saponin and reaches 83.4%, and total Saponin rate of transform is 81.2%.
Embodiment 4
Take by weighing Milkvetch Root 20kg, pulverize, cross 20 mesh sieves, add 50% ethanol of 12 times of amounts, 90 ℃ are extracted each 1.5 hours 3 times.Merge extractive liquid,, get extractum, with this extractum with an amount of dissolved in distilled water, last macroporous resin column respectively with distilled water, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 98% ethanol elution, merges the 40%-98% ethanol elution, evaporate to dryness gets extract I, calculate with astragaloside, this extract contains total Saponin and reaches 74.8%, and total Saponin rate of transform is 85.1%.
Claims (5)
1, a kind of astragaloside, available following method obtains:
1) get the ethanol extraction extractum of the Radix Astragali, use water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, evaporate to dryness, the Radix Astragali total saponins crude product;
2) or, get the ethanol extraction extractum of the Radix Astragali, use water dissolution, earlier with the saturated n-butanol extraction of alkaline solution, concentrated extract is to the extractum shape, reuse water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution, merge the 70%-98% ethanol elution, evaporate to dryness gets the Radix Astragali total saponins crude product;
3) or, get the ethanol extraction extractum of the Radix Astragali, use dissolve with methanol, add kieselguhr and mix thoroughly, extract with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness, the Radix Astragali total saponins crude product;
With the astragaloside crude product that obtains, handle through following method:
With 1) in obtain the Radix Astragali total saponins crude product, use water dissolution, go up macroporous resin column once more, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, evaporate to dryness, Radix Astragali total saponins;
Perhaps, with 1) in obtain the Radix Astragali total saponins crude product, use dissolve with methanol, add kieselguhr and mix thoroughly, extract with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness, Radix Astragali total saponins;
Perhaps, with 2) in obtain the Radix Astragali total saponins crude product, use dissolve with methanol, add kieselguhr and mix thoroughly, extract with the dehydrated alcohol hot reflux, gained extracting solution evaporate to dryness, Radix Astragali total saponins;
Perhaps, with 3) in obtain the Radix Astragali total saponins crude product, use water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, evaporate to dryness, Radix Astragali total saponins;
Perhaps, with 3) in obtain the Radix Astragali total saponins crude product, use water dissolution, with the saturated n-butanol extraction of NaOH of 0.1N, concentrated extract is to the extractum shape earlier, the reuse water dissolution, last macroporous resin column, water and aquiferous ethanol gradient elution merge the 70%-98% ethanol elution, evaporate to dryness gets Radix Astragali total saponins.
2, according to the described astragaloside of claim 1, it is characterized in that: the model of macroporous resin column is D101, AB-8; The alkaline solution that the n-butyl alcohol that is used to extract is saturated is the NaOH of 0.1N.
3, according to the described astragaloside of claim 1, it is characterized in that: ethanol extraction extractum and diatomaceous weight ratio are 1: 1-3.
4, according to the described astragaloside of claim 3, it is characterized in that: ethanol extraction extractum and diatomaceous weight ratio are for being 1: 2.
5, be used to prevent and treat the pharmaceutical composition of cardiovascular disease, wherein contain the astragaloside and the pharmaceutically acceptable carrier of the claim 1 for the treatment of effective dose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031133304A CN1199651C (en) | 2003-04-29 | 2003-04-29 | Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031133304A CN1199651C (en) | 2003-04-29 | 2003-04-29 | Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1444949A CN1444949A (en) | 2003-10-01 |
| CN1199651C true CN1199651C (en) | 2005-05-04 |
Family
ID=27814704
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB031133304A Expired - Fee Related CN1199651C (en) | 2003-04-29 | 2003-04-29 | Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100434093C (en) * | 2004-02-11 | 2008-11-19 | 吴芳 | 'Kidney-Benefiting' glucose injection and its preparing process |
| CN100459987C (en) * | 2004-02-23 | 2009-02-11 | 江苏省药物研究所 | Effective part of astragalus, preparing method and application in pharmacy |
| CN1785202B (en) * | 2004-12-06 | 2010-12-01 | 天津天士力制药股份有限公司 | Medicinal composition for treating cardiovascular disease and its preparation method |
| CN100441203C (en) * | 2005-02-24 | 2008-12-10 | 四川科伦药业股份有限公司 | Method for preparing astragalus root saponin |
| CN102133262A (en) * | 2011-03-13 | 2011-07-27 | 浙江大学 | Preparation method and application of Astragalus membranaceus active constituent with effect of myocardium protection |
-
2003
- 2003-04-29 CN CNB031133304A patent/CN1199651C/en not_active Expired - Fee Related
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