CN102558259A - Method for extracting purified complanatuside from milk vetch seed - Google Patents
Method for extracting purified complanatuside from milk vetch seed Download PDFInfo
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- CN102558259A CN102558259A CN2010106039204A CN201010603920A CN102558259A CN 102558259 A CN102558259 A CN 102558259A CN 2010106039204 A CN2010106039204 A CN 2010106039204A CN 201010603920 A CN201010603920 A CN 201010603920A CN 102558259 A CN102558259 A CN 102558259A
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- complanatoside
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- astragali complanati
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Abstract
The invention discloses a method for extracting purified complanatuside from milk vetch seed. The method comprises the following steps of: extracting milk vetch seed with alkaline water or an alkaline diluted alcohol solution; filtering; adding an extracting solution into an ultrafiltration membrane system; collecting a permeate; adjusting the pH of the permeate to 3-5; dissolving precipitates with hot water; cooling; removing fat-soluble impurities with petroleum ether; adding a precipitating agent into a water layer for clarifying; and adsorbing a liquid with macroporous resin, separating and purifying with a medium-pressure silica gel column and recrystallizing with chloroform-methanol to obtain a high-purity product, wherein the purity of the obtained product can be over 98 percent. In the method, a membrane separating technology is adopted, so that the purity of complanatuside is increased, the damages of resin are reduced, and the service life of resin is prolonged; and moreover, the production period of the medium-pressure silica gel column is short; and most edible ethanol is used in extracting and purifying processes, so that pollution is low.
Description
Technical field:
The invention belongs to the Natural Medicine Chemistry technical field, be specifically related to a kind of method of from Semen Astragali Complanati, extracting the purifying complanatoside.
Background technology:
Semen Astragali Complanati derives from the dry mature seed of leguminous plants astragatus complanatus Astragalus complanatus R.Br..Sweet, temperature.Return liver, kidney channel.Have and tonify the kidney and support yang, the controlling nocturnal emission with astringent drugs urine that contracts, the effect that nourishes the liver to improve visual acuity.Be used for lumbago due to renal deficiency, the seminal emission premature ejaculation, enuresis frequent micturition, dizzy under the gonorrhoea band, order is secretly dim-sighted.There is the bibliographical information Semen Astragali Complanati all influential, and also has the effect of antipyretic-antalgic, calmness, cold-resistant, antifatigue, anti-inflammatory and fat-reducing liver-protecting hemorheology, immunologic function, blood pressure and cerebral blood flow (CBF).The contained chemical ingredients of Semen Astragali Complanati mainly contains amino acid, flavonoid, triterpenes, lipid acid and trace element etc.Wherein flavones is one of main effective constituent of Semen Astragali Complanati, has many pharmacologically actives.Pharmacological research shows that semen astragali complanati general flavone has tangible reduction serum cholesterol, triglyceride, alleviates the effect of hepatic steatosis shape, also can reduce lipoprotein-cholesterol level.Recent study thinks that semen astragali complanati general flavone can protect the rat vitals to avoid sport injury, obviously improves the motor capacity of rat, suppresses hepatic fibrosis and forms, and antitumor E4-63 has tangible hypotensive effect to SHR and RHR-83 in addition.Complanatoside A is the principal character property composition of Semen Astragali Complanati flavones, and the content in Semen Astragali Complanati is higher, also is that Semen Astragali Complanati protects the liver one of main effective constituent of anti-hepatic fibrosis.
Wu Wenqian etc. have introduced the preparation method of complanatoside A reference substance in document " preparation and the evaluation of complanatoside A reference substance in the Semen Astragali Complanati ", this method is got Semen Astragali Complanati meal petroleum ether degreasing, 80% extraction using alcohol; ZTC-5 flavones absorption with macroporous adsorbent resin, 50% ethanol elution is through polyamide column chromatography; Water and different concentration ethanol gradient elution; Go up silicagel column again, use vinyl acetic monomer: the methyl alcohol gradient elution, vinyl acetic monomer: complanatoside A crystallization is separated out at methyl alcohol (10: 2) wash-out position; Methyl alcohol carries out repeatedly recrystallization, gets faint yellow needle crystal.This method is through three column chromatographies, and the production cycle is long, loses greatlyyer, and the recovery is low, is difficult to enlarge produce.
Application number is the preparation method that 200910057073.3 Chinese patent has been introduced a kind of complanatoside A, and this method adopts 80% extraction using alcohol, ethyl acetate extraction, water-saturated n-butanol extraction, uses macroporous resin adsorption then; 10-30% methyl alcohol, 50-70% methanol-eluted fractions, 50-70% ethanol elution partially concd is to doing dissolve with methanol; Last anti-phase C18 post; The 30-50% methanol-eluted fractions is concentrated into dried, recrystallization and promptly gets, and purity is more than 98%.This method is used a large amount of organic solvents and deleterious methanol solution, and dissolvent residual is bigger.
Summary of the invention:
The object of the present invention is to provide a kind of method of from Semen Astragali Complanati, extracting the purifying complanatoside, especially adopt membrane filtration technology, macroporous resin adsorption, medium pressure column chromatography method to prepare the method for high purity complanatoside.
The objective of the invention is to realize through following technical scheme:
A kind of method of from Semen Astragali Complanati, extracting the purifying complanatoside is characterized in that may further comprise the steps: get Semen Astragali Complanati and extract with alkaline water or alkaline dilute alcohol solution, filter, extracting solution adds ultrafiltration membrane system; Collect and see through liquid, add acid again after concentrating through liquid and transfer pH to 3-5, deposition adds the hot water dissolving, puts cold; With petroleum ether extraction 2-4 time, weeding of grease solubility impurity, water layer adds the precipitation agent clarification; Liquid is through macroporous resin adsorption, the 40-70% ethanol elution, and elutriant is condensed into medicinal extract; Add the silica gel medium pressure column separating purification, the ethyl acetate-ethanol gradient elution, chloroform-methanol (3: 1) recrystallization promptly gets complanatoside.
Described alkaline water is ammonia soln, sodium hydroxide solution, sodium hydrogen carbonate solution, liming, and alcohol is ethanol in the described alkaline dilute alcohol solution, and concentration is 40-60%, and the liquid material is than being 8-20: 1.
Described process for extracting is heating reflux method, supersound extraction, diacolation extraction, microwave extraction or decocting method.
Described ultra-filtration membrane is the tubular type composite hyperfiltration membrane of molecular weight cut-off 4000-10000.
The 4-8 that described sherwood oil add-on is a precipitation volume doubly.
Described precipitation agent is ZTC finings, CTS Chinese medicine finings, remove in the acetyl chitosan any, and add-on is the 0.6%-2.2% of liquid.
Described macroporous resin is a kind of among DS401, HZ818, LS-303, the ADS-21, and the eluent consumption is a 4-8 times of column volume.
Described silica gel medium pressure post column length is 50-300cm, and column diameter is 5-100cm, and filling gel order number is the 100-300 order, and post is pressed and is 0.1-4MPa.
Described ethyl acetate-ethanol mixed solvent ratio is 20: 1-2: 1.
Beneficial effect of the present invention: production cost is low, with short production cycle, product purity is high, the agents useful for same hypotoxicity, and industrial continuous production is convenient in recyclable utilization.
Embodiment:
Embodiment 1:
Get Semen Astragali Complanati raw material 5kg and drop in the small-sized extractor, add 8 times of amount 2% sodium hydroxide solution diacolations and be extracted into the percolate color and shoal, merge percolate, add the tubular type composite hyperfiltration membrane system ultrafiltration of molecular weight cut-off 6000; See through liquid and be condensed into and add hydrochloric acid behind the medicinal extract and transfer pH to 3.5, leach deposition, the dissolving of throw out reflux is put cold; With 5 times of volume petroleum ether extractions 2 times, water layer adds 2.2%CTS Chinese medicine finings, leaves standstill; Centrifugal, HZ818 macroporous adsorptive resins on the clear liquid is earlier with the washing decon; With 50% ethanol elution, collect elutriant again, be condensed into medicinal extract.With 100-200 order silica gel 4kg with the ETHYLE ACETATE suspendible after, evenly add column length 200cm, in the stainless steel chromatography post of internal diameter 20cm, drive away bubble; Pressure-controlling is disperseed 200g medicinal extract at 0.3MPa with ETHYLE ACETATE, use the pump sample introduction, and last appearance is back with ethyl acetate-ethanol 20: 1-2: 1 gradient elution; Fractional Collections, TLC tracking monitor, the flow point of merging high-content complanatoside; Reclaim under reduced pressure reagent, chloroform-methanol (3: 1) recrystallization leaches crystallization; Drying gets complanatoside 2.5g, purity 98.4%.The eluent that reclaims can be done to reuse next time after regeneration.
Embodiment 2:
Get Semen Astragali Complanati raw material 5kg and drop in the small-sized extractor, add 15 times of amount 3% lime aqueous solution supersound extraction 4 times, each 1 hour, united extraction liquid; The tubular type composite hyperfiltration membrane system ultrafiltration that adds molecular weight cut-off 4000 sees through liquid and is condensed into and adds hydrochloric acid behind the medicinal extract and transfer pH to 3.0, leaches deposition, the dissolving of throw out reflux; Put coldly, with 4 times of volume petroleum ether extractions 3 times, water layer adds 1.5% and removes the acetyl chitosan, leaves standstill; Centrifugal, ADS-21 macroporous adsorptive resins on the clear liquid is earlier with the washing decon; With 70% ethanol elution, collect elutriant again, be condensed into medicinal extract.With 200-280 order silica gel 6kg with the ETHYLE ACETATE suspendible after, evenly add column length 100cm, in the stainless steel chromatography post of internal diameter 5cm, drive away bubble, pressure-controlling is at 0.8MPa; 200g medicinal extract is disperseed with ETHYLE ACETATE, use the pump sample introduction, last appearance is back with ethyl acetate-ethanol 20: 1-2: 1 gradient elution, Fractional Collections; The TLC tracking monitor, the flow point of merging high-content complanatoside, reclaim under reduced pressure reagent, chloroform-methanol (3: 1) recrystallization; Leach crystallization, drying gets complanatoside 2.3g, purity 98.7%.The eluent that reclaims can be done to reuse next time after regeneration.
Embodiment 3:
Get Semen Astragali Complanati raw material 10kg and drop in the small-sized extractor, add 20 times of amount 5% ammoniacal liquor alcoholic solutions (determining alcohol is 50%) and decoct, filter united extraction liquid; The tubular type composite hyperfiltration membrane system ultrafiltration that adds molecular weight cut-off 5000 sees through liquid and is condensed into and adds hydrochloric acid behind the medicinal extract and transfer pH to 4.0, leaches deposition, the dissolving of throw out reflux; Put coldly, with 8 times of volume petroleum ether extractions 2 times, water layer adds 1.2%CTS Chinese medicine finings, leaves standstill; Centrifugal, LS-303 macroporous adsorptive resins on the clear liquid is earlier with the washing decon; With 40% ethanol elution, collect elutriant again, be condensed into medicinal extract.With 150-250 order silica gel 10kg with the ETHYLE ACETATE suspendible after, evenly add column length 250cm, in the stainless steel chromatography post of internal diameter 50cm, drive away bubble; Pressure-controlling is disperseed 400g medicinal extract at 2.3MPa with ETHYLE ACETATE, use the pump sample introduction, and last appearance is back with ethyl acetate-ethanol 20: 1-2: 1 gradient elution; Fractional Collections, TLC tracking monitor, the flow point of merging high-content complanatoside; Reclaim under reduced pressure reagent, chloroform-methanol (3: 1) recrystallization leaches crystallization; Drying gets complanatoside 5.0g, purity 98.2%.The eluent that reclaims can be done to reuse next time after regeneration.
Embodiment 4:
Get Semen Astragali Complanati raw material 10kg and drop in the small-sized extractor, add 16 times of amount 3% sodium hydrogencarbonate alcoholic solution (determining alcohol is 40%) microwave extraction 3 times, each 45 minutes, united extraction liquid; The tubular type composite hyperfiltration membrane system ultrafiltration that adds molecular weight cut-off 8000 sees through liquid and is condensed into and adds hydrochloric acid behind the medicinal extract and transfer pH to 4.5, leaches deposition, the dissolving of throw out reflux; Put coldly, with 6 times of volume petroleum ether extractions 3 times, water layer adds the 0.6%ZTC1+1 finings, leaves standstill; Centrifugal, DS401 macroporous adsorptive resins on the clear liquid is earlier with the washing decon; With 60% ethanol elution, collect elutriant again, be condensed into medicinal extract.With 300-400 order silica gel 8kg with the ETHYLE ACETATE suspendible after, evenly add column length 300cm, in the stainless steel chromatography post of internal diameter 80cm, drive away bubble; Pressure-controlling is disperseed 400g medicinal extract at 3.6MPa with ETHYLE ACETATE, use the pump sample introduction, and last appearance is back with ethyl acetate-ethanol 20: 1-2: 1 gradient elution; Fractional Collections, TLC tracking monitor, the flow point of merging high-content complanatoside; Reclaim under reduced pressure reagent, chloroform-methanol (3: 1) recrystallization leaches crystallization; Drying gets complanatoside 4.9g, purity 98.5%.The eluent that reclaims can be done to reuse next time after regeneration.
Claims (9)
1. a method of from Semen Astragali Complanati, extracting the purifying complanatoside is characterized in that may further comprise the steps: get Semen Astragali Complanati and extract with alkaline water or alkaline dilute alcohol solution, filter, extracting solution adding ultrafiltration membrane system; Collect and see through liquid, add acid again after concentrating through liquid and transfer pH to 3-5, deposition adds the hot water dissolving, puts cold; With petroleum ether extraction 2-4 time, weeding of grease solubility impurity, water layer adds the precipitation agent clarification; Liquid is through macroporous resin adsorption, the 40-70% ethanol elution, and elutriant is condensed into medicinal extract; Add the silica gel medium pressure column separating purification, the ethyl acetate-ethanol gradient elution, chloroform-methanol (3: 1) recrystallization promptly gets complanatoside.
2. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1; It is characterized in that; Described alkaline water is ammonia soln, sodium hydroxide solution, sodium hydrogen carbonate solution, liming; Alcohol is ethanol in the described alkaline dilute alcohol solution, and concentration is 40-60%, and the liquid material is than being 8-20: 1.
3. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that described process for extracting is heating reflux method, supersound extraction, diacolation extraction, microwave extraction or decocting method.
4. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that described ultra-filtration membrane is the tubular type composite hyperfiltration membrane of molecular weight cut-off 4000-10000.
5. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that, the 4-8 that described sherwood oil add-on is a precipitation volume doubly.
6. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that, described precipitation agent is ZTC finings, CTS Chinese medicine finings, remove in the acetyl chitosan any, and add-on is the 0.6%-2.2% of liquid.
7. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that, described macroporous resin is a kind of among DS401, HZ818, LS-303, the ADS-21, and the eluent consumption is a 4-8 times of column volume.
8. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that described silica gel medium pressure post column length is 50-300cm, and column diameter is 5-100cm, and filling gel order number is the 100-300 order, and post is pressed and is 0.1-4MPa.
9. the method for from Semen Astragali Complanati, extracting the purifying complanatoside as claimed in claim 1 is characterized in that described ethyl acetate-ethanol mixed solvent ratio is 20: 1-2: 1.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010106039204A CN102558259A (en) | 2010-12-24 | 2010-12-24 | Method for extracting purified complanatuside from milk vetch seed |
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| CN2010106039204A CN102558259A (en) | 2010-12-24 | 2010-12-24 | Method for extracting purified complanatuside from milk vetch seed |
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| CN102558259A true CN102558259A (en) | 2012-07-11 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015021632A1 (en) * | 2013-08-15 | 2015-02-19 | 山西振东制药股份有限公司 | Astragalus membranaceus seed extract and use thereof for preparing animal feed and foods |
| CN110960571A (en) * | 2018-09-29 | 2020-04-07 | 上海中医药大学附属龙华医院 | Preparation method of semen astragali complanati total flavonoids |
| CN113368122A (en) * | 2021-07-02 | 2021-09-10 | 五邑大学 | Application of complanatoside in preparing medicine for treating skin inflammation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1544011A (en) * | 2003-11-26 | 2004-11-10 | 苏州中药研究所 | Application of flatstem milkvetch seed and its extract in the preparation of anti hepatic fibrosis medicine and anticancer medicine |
| US20090252796A1 (en) * | 2008-04-07 | 2009-10-08 | Mazed Mohammad A | Nutritional supplement for the prevention of cardiovascular disease, alzheimer's disease, diabetes, and regulation and reduction of blood sugar and insulin resistance |
| CN101863939A (en) * | 2009-04-15 | 2010-10-20 | 上海诗丹德生物技术有限公司 | Method for preparing complanatoside A |
-
2010
- 2010-12-24 CN CN2010106039204A patent/CN102558259A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1544011A (en) * | 2003-11-26 | 2004-11-10 | 苏州中药研究所 | Application of flatstem milkvetch seed and its extract in the preparation of anti hepatic fibrosis medicine and anticancer medicine |
| US20090252796A1 (en) * | 2008-04-07 | 2009-10-08 | Mazed Mohammad A | Nutritional supplement for the prevention of cardiovascular disease, alzheimer's disease, diabetes, and regulation and reduction of blood sugar and insulin resistance |
| CN101863939A (en) * | 2009-04-15 | 2010-10-20 | 上海诗丹德生物技术有限公司 | Method for preparing complanatoside A |
Non-Patent Citations (2)
| Title |
|---|
| 张建军，等: "HPLC 测定沙苑子中沙苑子苷A的含量", 《中国中药杂志》 * |
| 缪勇，等: "《中草药植物提取与深加工新技术实用手册》", 31 December 2004, 天津电子出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015021632A1 (en) * | 2013-08-15 | 2015-02-19 | 山西振东制药股份有限公司 | Astragalus membranaceus seed extract and use thereof for preparing animal feed and foods |
| CN110960571A (en) * | 2018-09-29 | 2020-04-07 | 上海中医药大学附属龙华医院 | Preparation method of semen astragali complanati total flavonoids |
| CN113368122A (en) * | 2021-07-02 | 2021-09-10 | 五邑大学 | Application of complanatoside in preparing medicine for treating skin inflammation |
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Application publication date: 20120711 |