CN100441203C - Method for preparing astragalus root saponin - Google Patents
Method for preparing astragalus root saponin Download PDFInfo
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- CN100441203C CN100441203C CNB2005100204029A CN200510020402A CN100441203C CN 100441203 C CN100441203 C CN 100441203C CN B2005100204029 A CNB2005100204029 A CN B2005100204029A CN 200510020402 A CN200510020402 A CN 200510020402A CN 100441203 C CN100441203 C CN 100441203C
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Abstract
The present invention relates to a method for preparing astragaloside for injections. After astragalus root is extracted by alcohol containing water by backflow, an extracting solution of the ethanol is removed, and the alcohol is used for precipitation and filtration; the alcohol is removed, and water is added for cryoprecipitation; filter liquor is absorbed by adsorptive resin and is eluted by the alcohol containing water; after being concentrated and regulated to alkalin, the eluent is extracted by n-butanol saturated by water; finally, the extraction liquid without the n-butanol is precipitated and separated by acetone or ether to obtain an astragaloside precipitate for injections. The method can thoroughly remove large amount of phenol and acidic substances in the astragalus root and effectively ensures the qualification and the stability of various indexes of the injection prepared from astragaloside products for injections, such as clarity, color, tannin, resin, oxalate, etc.
Description
Technical field
What the present invention relates to is the preparation method of the Radix Astragali saponin used for injection.
Background technology
The Radix Astragali is traditional simply qi-invigorating herb, and existing pharmacological research shows that main effective ingredient Radix Astragali saponin wherein can obviously improve the myocardial contractility of myocardial infarction dog; increase coronary flow; dwindle myocardial infarction area, alleviate myocardial damage, cardiac function is had protective effect.There are some researches show that the main effective ingredient in the Radix Astragali is a Radix Astragali saponin, but content is lower, particularly the content of astragaloside composition in medical material wherein only is about 0.04%, adopts present traditional extraction process to make comparatively difficulty of its enriching and purifying.
Publication number is the extracting method that report has pair Radix Astragali saponin in the Chinese patent literature of CN1557329A, what adopt is the aqueous extract of producing the raw material crude drug Radix Astragali earlier in the conventional decocting mode of boiling, filtrate is 0.02～0.03 μ m through macroporous resin commonly used as average pore size, specific surface area 〉=400m
2The macroporous adsorbent resin of models such as the D-201 of/g, AB-8, DM-301 carries out adsorption treatment, adsorption and enrichment saponin constituent wherein.After water and the remove impurity of aquiferous ethanol solution washing, alcoholic solution desorbing eluting with 60%～95%, remove the eluent water dissolution behind the ethanol, with n-butanol extraction Radix Astragali total saponins wherein, and with sodium hydroxide/potassium, sodium carbonate, the soda lye wash n-butanol extraction of pH such as saturated limewater or ammonia 〉=10 is molten, remove reuse acetone backflow defat behind the n-butyl alcohol, can obtain the Radix Astragali saponin of Astragaloside content 〉=20% (weight) after the separation, and can be by " pharmaceutical analysis magazine " 1999,19 (6): 403～405 and " Chinese Pharmaceutical Journal " 2002,37 (4): 298～300 report, with high performance liquid chromatography and evaporative light scattering detector, promptly the HPLC-ELSD method is measured the Astragaloside content in the resulting Radix Astragali saponin extract.The result of the test demonstration, simple low with the rate of transform of water extraction Radix Astragali saponin, and the gained Radix Astragali saponin all can not meet the requirement of injectable drug on many indexs such as clarity, tannin.
Summary of the invention
At above-mentioned situation, the present invention will provide a kind of preparation method of Radix Astragali saponin of injection, make it can remove a large amount of phenol, acidic materials and tannin etc., can keep the stable of Radix Astragali saponin again, satisfy the required performance indications of using for injection of Radix Astragali saponin, guarantee the safety of its use.
The preparation method of astragalus root saponin of the present invention is to be 40%～95% with medicine material Radix Astragali volumetric concentration, is more preferably the aquiferous ethanol reflux, extract, with 60%～80%.Result of the test shows that it is low excessively to extract used concentration of alcohol, and the water-solubility impurity in the extract is too much; Then can increase the oil-soluble impurities in the extract when concentration of alcohol is too high again too much.Compare with the water extraction mode of routine, extract Radix Astragali saponin with 40～95% ethanol circulating refluxs, the rate of transform of Radix Astragali saponin is higher, and the rate of transform can reach more than 92%, and only be 71% with the rate of transform of water extraction mode merely, the two exists notable difference (P＜0.01).
Reflux extracting liquid is removed ethanol, adds alcohol volume content to 40%～95% of ethanol to mixed solution again, is more preferably to reach 70%～90% to ethanol content and precipitate and filter.Result of the test shows, filtrate is adjusted to pH 6.0～11.0 after, can remove tannin effectively, the scope that wherein preferably is adjusted to pH 8.0～10.0 is carried out sedimentation and filtration.After filtrate is adjusted to pH 6.0～8.0, it by crude drug/water weight ratio 0.5～3/1 amount, wherein being more preferably by crude drug/water is after 1～2/1 amount adds water and mixes, keeping uncongealable cryogenic conditions, wherein preferably to precipitate after-filtration under 0 ℃～10 ℃ conditions.Filtrate is used macroporous adsorbent resin commonly used in the pharmaceutical manufacturing, as average pore size is 0.02～0.03 μ m, the macroporous adsorbent resin of the D-101 commonly used of specific surface area 〉=400m2/g, Diaion HP-20, D-201, models such as AB-8, DM-301 carries out adsorption treatment, and reuse concentration is 50%～90% aquiferous ethanol desorption eluting and collects.Result of the test shows that used concentration of alcohol is low excessively during eluting, can be mixed with too much water-solubility impurity in the eluent; The too high amount that then may increase oil-soluble impurities in the eluent of concentration of alcohol.With after removing desorption concentrated solution behind the ethanol and regulating pH to 9.0～13.0, with through water saturated n-butanol extraction, extracting solution is removed at least a precipitate and separate precipitate that carries out in the acetone and ether behind the n-butyl alcohol, is the said Radix Astragali saponin of using for injection of the present invention.
Experiment shows, in the above-mentioned preparation method, handle and be adjusted to the extracting solution of nearly neutral pH 6.0～8.0 through ethanol precipitation, and further add water and mix and be incorporated in precipitation process under the low temperature, the clarity of astragalus root saponin solution all is important and necessary when guaranteeing to use.Result of the test shows, before adding water and mixing the meta-acid of pH regulator or partially alkali all can produce significantly adverse effect to the content of astragaloside.And no longer relatively poor through the sample clarity of water precipitation remove impurity processing, the sample clarity that the remove impurity of process water precipitation is handled then can meet the standard-required of injection.
Getting and be equivalent to the concentrated solution of 800g medical material through ethanol precipitation, is that different proportion such as 1/1,1.5/1,2/1 and 2.5/1 adds water to carry out the result of the test that cold preservation staticly settles as shown in table 1 by Milkvetch Root/water respectively.
By table 1 result as seen: the sample clarity after the aqueous precipitation remove impurity is handled all can meet the standard-required of injection, change the water yield what treatment effect is not seen appreciable impact, but in operation, different influences is arranged then.From the water precipitating liquid rate of filtration, medical material/water is 1/1 and 1.5/1 o'clock, and the rate of filtration is very fast, and when medical material/water is 2.0/1 and 2.5/1, filters then relatively difficulty.The adding water yield when therefore carrying out the water precipitation processing, generally the ratio with medical material/water is 1～2/1, is good with 1～1.5/1 o'clock particularly.
When the extracting solution continuation after water precipitation is handled is handled with macroporous resin adsorption, change the kind and/or the model of adsorbent resin, as the evidence of selecting at multiple nonpolar macroporous adsorption resins such as above-mentioned D-101, Diaion HP-20, D-201, AB-8, DM-301 and/or low pole adsorbent resin macroporous adsorbent resin, it all can use, and the quality of products obtained therefrom and stability etc. can both meet the requirements.Further, when carrying out adsorption treatment with macroporous resin, before washing the desorption eluting with 50%～90% aquiferous ethanol, if can be colourless substantially with 20%～50% aquiferous ethanol washing adsorbent resin earlier to cleaning mixture, then to the impurity in the further minimizing collection eluted product, it is useful improving product purity and quality.Test shows, tentatively washs concentration of alcohol that impurity uses and crosses and lowly can make the impurity level in extract obtained many; The too high yield that then can influence Radix Astragali saponin in the extract of concentration of alcohol.
A large amount of and the main impurity component that exists in the Milkvetch Root is approaching phenol of polarity and Radix Astragali saponin and acidic materials, removes relatively difficulty.Therefore in the above-mentioned preparation method, before with the water saturation n-butanol extraction, extracting solution after desorption concentrated is regulated pH to alkalescence with alkali commonly used or alkali compounds, can make phenol and acid impurities salify, be insoluble in n-butyl alcohol and be removed, thereby all be important for quality that guarantees final products and stability.Said alkali or alkali compounds generally can use hydroxide, carbonate or the bicarbonate of monovalence metal commonly used, Chang Yong sodium hydroxide, sodium carbonate, sodium bicarbonate the most, or alkali or alkali compounds such as ammonia.This goes on foot preferred pH value range of accommodation is 10.0～12.0, and then uses water saturated n-butanol extraction.Test shows that the kind that changes used alkali when regulating pH there is no substantial influence to the result.Extracting solution after handling through absorption with macroporous adsorbent resin respectively with using the water saturation n-butanol extraction complete respectively behind the different alkali adjusting pH, is tested the retention rate of Radix Astragali saponin and astragaloside.Result of the test is as shown in table 2.
The result of table 2 shows that the extracting solution before the extraction is handled with different alkali or alkali compounds, and the retention rate that extracts the back Radix Astragali saponin is not had significant difference.With relative more weak ammonia and the NaHCO of basicity
3During solution-treated, the Astragaloside content in the extract is on the low side relatively, and with after 0.5% and 1% the NaOH solution-treated, the retention rate of astragaloside obviously improves in the extract.Use 1%Na
2CO
3During solution-treated, can improve the content of astragaloside, and the color of alkaline mixed solution is also more shallow after handling.But it is strong always favourable that to be not meaning alkalescence cross for this, may have adverse effect to extract obtained dissolubility by force because alkalescence is crossed.Therefore can preferentially select Na for use
2CO
3Regulate pH.Directly use the test of water saturation n-butanol extraction to show without above-mentioned alkali treatment to the concentrated solution before extracting, the Radix Astragali saponin solution of gained all is difficult to reach the requirement of the technical specification and the regulation of relevant Chinese medicine injection at clarity and color, tannin, resin, oxalates etc., can not be as the injection raw material.With without alkali treatment directly the sample that obtains of extraction the Cavia porcellus test then idol anaphylaxis is arranged, react and all has allergy with the Cavia porcellus test that resulting sample carries out after alkali treatment before the extraction.Simultaneously, the investigation of stability test shows that also with the injection that alkali-treated raw material is made, its safety all met the requirements at least in 2 years.Therefore, carry out alkali treatment before the extraction and positive effect is arranged guaranteeing product aspect such as safety and stability when being used for injection.
Be appreciated that thus preparation method of the present invention at first adopts aquiferous ethanol to extract Radix Astragali saponin, can obtain the higher Radix Astragali saponin rate of transform; By repeatedly Ethanol Treatment, alkali treatment and precipitation process to extracting solution, the a large amount of phenol and the acidic materials that exist in the Milkvetch Root are removed comparatively completely, guaranteed the clarity of astragalus root saponin product and standard and the requirement that every indexs such as color, tannin, resin, oxalates and stability all can reach injection effectively.Steps such as particularly multi-form alkali treatment and water at low temperature precipitation remove impurity guarantee especially and improve the important measures that the Radix Astragali saponin product reaches injecting drug use standard and requirement.
The water precipitating comparative test result of the different water consumptions of table 1
The comparative test result of the alkali treatment condition that table 2 is different
Below example by the specific embodiment again foregoing of the present invention is described in further detail.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.Do not breaking away under the above-mentioned technological thought situation of the present invention, various replacements or change according to ordinary skill knowledge and customary means are made include within the scope of the invention.
The specific embodiment
Embodiment 1
Get Milkvetch Root 20kg, section adds volumetric concentration and is 8 times of reflux, extract, of aquiferous ethanol of 70%.Extracting solution is removed ethanol, adds ethanol to the alcohol volume content to 65% of mixed solution again and precipitates and filter, and filtrate is adjusted to the pH9.0 postprecipitation and filters.Filtrate is adjusted to about pH7.0, is after 1.5/1 amount adds water and mixes, to precipitate after-filtration under 5 ℃ of low temperature by crude drug/water.Filtrate is with macroporous adsorbent resin D-101 absorption, is eluted to the almost colourless remove impurity of eluent with the aquiferous ethanol of 30% concentration, again with the aquiferous ethanol desorption eluting of 75% concentration and collect.With after removing desorption concentrated solution behind the ethanol and regulating pH to 10.0, to use through water saturated n-butanol extraction, extracting solution precipitates with acetone-ether (1: 1) after removing n-butyl alcohol, the Radix Astragali saponin that sediment separate out is used for the confession injection.Press the injection standard test, the gained Radix Astragali saponin meets the injection requirement fully through full inspection.
Embodiment 2
Get Milkvetch Root 50kg, section adds volumetric concentration and is 8 times of reflux, extract, of aquiferous ethanol of 80%, and extracting solution is removed ethanol, adds ethanol to the alcohol volume content to 50% of mixed solution again and precipitates and filters, and filtrate is adjusted to the filtration of pH9.0 postprecipitation.Filtrate is adjusted to pH 7～8, is that 1.5/1 amount adds water and is mixed in precipitation after-filtration under 5 ℃ of low temperature by crude drug/water.Filtrate with macroporous adsorbent resin D-201 absorption after, it is almost colourless to be eluted to eluent with the aquiferous ethanol of 30% concentration earlier, again with the aquiferous ethanol desorption eluting and the collection of 80% concentration.With after removing desorption concentrated solution behind the ethanol and regulating pH to 11.0, to use through water saturated n-butanol extraction, extracting solution precipitates with acetone-ether (1: 2) after removing n-butyl alcohol, the Radix Astragali saponin that sediment separate out is used for the confession injection.The gained Radix Astragali saponin meets the injection requirement through full inspection.
Embodiment 3
Get Milkvetch Root 30kg, section adds volumetric concentration and is 10 times of reflux, extract, of aquiferous ethanol of 50%, and extracting solution is removed ethanol, adds ethanol to the alcohol volume content to 85% of mixed solution again and precipitates and filters, and filtrate is adjusted to the filtration of pH 9.0 postprecipitations.Filtrate is adjusted to pH 6.5～7.5, is that 1.8/1 amount adds water and is mixed in precipitation after-filtration under 5 ℃ of low temperature by crude drug/water.Filtrate with macroporous adsorbent resin D-101 absorption after, it is almost colourless to be eluted to eluent with the aquiferous ethanol of 30% concentration earlier, again with the aquiferous ethanol desorption eluting and the collection of 75% concentration.With after removing desorption concentrated solution behind the ethanol and regulating pH to 12.0, use through water saturated n-butanol extraction.Extracting solution precipitates with acetone-ether (2: 1) after removing n-butyl alcohol, the Radix Astragali saponin of sediment separate out for using for injection.The gained Radix Astragali saponin meets the injection requirement through full inspection.
Embodiment 4
Get Milkvetch Root 20kg, section adds volumetric concentration and is 8 times of reflux, extract, of aquiferous ethanol of 70%.Extracting solution is removed ethanol, adds ethanol to the alcohol volume content to 50% of mixed solution again and precipitates and filter, and filtrate is adjusted to the pH9.0 postprecipitation and filters.Filtrate is adjusted to about pH7.0, is after 1.5/1 amount adds water and mixes, to precipitate after-filtration under 5 ℃ of low temperature by crude drug/water.Filtrate is with macroporous adsorbent resin D-101 absorption, is eluted to the almost colourless remove impurity of eluent with the aquiferous ethanol of 30% concentration, again with the aquiferous ethanol desorption eluting of 75% concentration and collect.With after removing desorption concentrated solution behind the ethanol and regulating pH to 10.0, to use through water saturated n-butanol extraction, extracting solution precipitates with acetone after removing n-butyl alcohol, the Radix Astragali saponin of sediment separate out for using for injection.The gained Radix Astragali saponin meets the injection requirement through full inspection.
Embodiment 5
Get Milkvetch Root 50kg, section adds volumetric concentration and is 8 times of reflux, extract, of aquiferous ethanol of 60%, and extracting solution is removed ethanol, adds ethanol to the alcohol volume content to 60% of mixed solution again and precipitates and filters, and filtrate is adjusted to the filtration of pH9.0 postprecipitation.Filtrate is adjusted to pH 7～8, is that 1.5/1 amount adds water and is mixed in precipitation after-filtration under 5 ℃ of low temperature by crude drug/water.Filtrate with macroporous adsorbent resin D-201 absorption after, it is almost colourless to be eluted to eluent with the aquiferous ethanol of 30% concentration earlier, again with the aquiferous ethanol desorption eluting and the collection of 80% concentration.With after removing desorption concentrated solution behind the ethanol and regulating pH to 11.0, to use through water saturated n-butanol extraction, extracting solution precipitates with ether after removing n-butyl alcohol, the Radix Astragali saponin of sediment separate out for using for injection.The gained Radix Astragali saponin meets the injection requirement through full inspection.
Claims (8)
1. the preparation method of astragalus root saponin, it is characterized in that with medicine material Radix Astragali volumetric concentration being 40%～95% aquiferous ethanol reflux, extract,, extracting solution is removed ethanol, again adding ethanol to alcohol volume content to 40%～95% of mixed solution precipitates and filters, filtrate is adjusted to pH6.0～11.0 postprecipitations and filters, after filtrate is adjusted to pH6.0～8.0, by crude drug/water weight ratio is that 1～2.5/1 amount adds water and mixes, keeping precipitating after-filtration under the uncongealable cryogenic conditions, filtrate is used absorption with macroporous adsorbent resin, aquiferous ethanol desorption eluting and collection with 50%～90% concentration, with after removing desorption concentrated solution behind the ethanol and regulating pH to 9.0～13.0, with through water saturated n-butanol extraction, extracting solution is removed behind the n-butyl alcohol with at least a precipitation the in acetone and the ether, the Radix Astragali saponin of sediment separate out for using for injection.
2. the preparation method of astragalus root saponin as claimed in claim 1 is characterized in that the concentration of the aquiferous ethanol that said Radix Astragali raw material reflux, extract, is used is 60%～80%.
3. the preparation method of astragalus root saponin as claimed in claim 1, it is characterized in that adding ethanol again by the said reflux extracting liquid of removing behind the ethanol, to carry out sedimentary mixed solution ethanol volumetric concentration be 70%～90%.
4. the preparation method of astragalus root saponin as claimed in claim 1 is characterized in that the said reflux extracting liquid of removing behind the ethanol adds after ethanol precipitates and filter again, is adjusted to pH8.0～10.0 sedimentation and filtrations with filtrate.
5. the preparation method of astragalus root saponin as claimed in claim 1, it is characterized in that extracting solution with macroporous resin adsorption after, colourless to washing liquid with the aquiferous ethanol washing adsorbent resin of 20%～50% concentration earlier, the aquiferous ethanol of reuse 50%～90% concentration is washed the desorption eluting.
6. the preparation method of astragalus root saponin as claimed in claim 1 is characterized in that the desorption concentrated solution of collecting and removing behind the ethanol is regulated pH10.0～12.0 with hydroxide, carbonate or the bicarbonate of monovalence metal or ammonia.
7. the preparation method of astragalus root saponin as claimed in claim 6 is characterized in that said monovalent metallic ion is sodium or potassium.
8. as the preparation method of the described astragalus root saponin of one of claim 1 to 7, it is characterized in that said filtrate is adjusted to pH6.0～8.0 after, be that 1～1.5/1 amount adds water and mixes by crude drug/water weight ratio.
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1844913B (en) * | 2006-03-22 | 2010-04-21 | 四川科伦药业股份有限公司 | Quality detection method for fingerprint spectrum of radix astragali saponin injection |
| CN101084982B (en) * | 2006-06-08 | 2011-09-21 | 天津天士力之骄药业有限公司 | Method for preparing total saponins of astragalus |
| CN101439079B (en) * | 2007-11-22 | 2012-04-18 | 天津天士力制药股份有限公司 | Preparation method of Chinese medicine granule containing total saponins in astragalus |
| CN101380353B (en) * | 2008-10-20 | 2012-02-01 | 东莞广州中医药大学中医药数理工程研究院 | Pharmaceutical composition for preventing and curing 2-type diabetes and preparation method thereof |
| CN102101880B (en) * | 2009-12-17 | 2013-06-12 | 北京大学 | Preparation method and application of radix astragali saponin |
| CN102241685B (en) * | 2010-05-11 | 2013-11-06 | 四川科伦药业股份有限公司 | Separation method of monomeric compound from astragaloside for injection |
| CN101912565B (en) * | 2010-05-25 | 2012-06-27 | 河南中医学院 | Turmeric and astragalus capsules for treating diabetes and incipient diabetic nephropathy |
| CN102232962B (en) * | 2011-07-25 | 2013-03-06 | 苏州大学 | Composition containing astragaloside active components as well as preparation method and application thereof |
| CN105560173B (en) * | 2015-12-29 | 2019-02-05 | 江苏九旭药业有限公司 | A kind of high stability astragalus injection and preparation method thereof |
| CN107334808B (en) * | 2017-08-23 | 2020-09-04 | 齐齐哈尔医学院 | Astragalus extract preparation with antibacterial activity |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1444949A (en) * | 2003-04-29 | 2003-10-01 | 中国药科大学 | Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease |
| CN1557329A (en) * | 2004-01-18 | 2004-12-29 | 成都厚发科技开发有限公司 | Medicine composition for treating cardiovascular and cerebrovascular disease and its prepararing method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1444949A (en) * | 2003-04-29 | 2003-10-01 | 中国药科大学 | Astragalus root valid part of Chinese herbal medicine for treating cardiovascular disease |
| CN1557329A (en) * | 2004-01-18 | 2004-12-29 | 成都厚发科技开发有限公司 | Medicine composition for treating cardiovascular and cerebrovascular disease and its prepararing method |
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Assignee: Liaoning Minkang Pharmaceutical Co.,Ltd. Assignor: Sichuan Kelun Pharmaceutical Co., Ltd. Contract fulfillment period: 2009.9.1 to 2019.8.31 contract change Contract record no.: 2009510000150 Denomination of invention: Method for preparing astragalus root saponin Granted publication date: 20081210 License type: Exclusive license Record date: 20091111 |
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