CN1470241A - Astragalus root methyl-glycoside composition and preparation method - Google Patents
Astragalus root methyl-glycoside composition and preparation method Download PDFInfo
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Abstract
The invention refers to astragaloside drug combination and the preparing method, the active component in the drug combination comes from triterpenoid saponis compound of Radix astragali, and the effective component in the compound is astragaloside IV. The method firstly uses the solvents like water to extract Radix astragali, and then makes separation and purification to obtain the coarse astragaloside, then makes hydrolyzation and recrystallization to obtain high-content astragaloside IV composition.
Description
Technical field
The present invention relates to a kind of astragalus root methyl-glycoside composition and preparation method thereof.
Background technology
The Radix Astragali is the dry root of leguminous plant Radix Astagali or the film folder Radix Astragali, and it is weak to be mainly used in the deficiency of vital energy clinically, anorexia and loose stool, sinking of QI of middle-JIAO, chronic diarrhea proctoptosis, the metrorrhagia of having blood in stool, the exterior deficiency spontaneous perspiration, deficiency of vital energy edema, carbuncle is difficult bursts, burst for a long time and do not hold back, blood deficiency fistula Huang, interior-heat is quenched one's thirst, chronic nephritis proteinuria, diabetes, cardiovascular and cerebrovascular disease, diseases such as antiviral, its main active are the triterpenoid saponin compounds based on astragaloside.
At present, the method for preparing Radix Astragali total saponins is, with Radix Astragali ethanol extraction, extracting solution concentrates that to reclaim ethanol complete, and residue adds the water suspendible, uses n-butanol extraction then, divide and get n-butyl alcohol liquid evaporate to dryness, Radix Astragali total saponins.Zhi Bei Radix Astragali total saponins by this method, its contained astragaloside has only 10-20%.In order to obtain high-load Radix Astragali total saponins, normally rough Radix Astragali total saponins is gone up silica gel or alumina column again, through solvent gradient elution, collect contain astragaloside the eluent evaporate to dryness promptly.Compare with the present invention, traditional method exists significantly not enough, and low as content, solvent consumption is big, complex process, and the cycle is long, and the cost height is not suitable for suitability for industrialized production.
Because the content of astragaloside is very low in the Radix Astragali, about 0.1%, be difficult to obtain high-load astragaloside compositions usually with common method.Have only by part Saponin hydrolysis in the Radix Astragali is converted into astragaloside, can obtain high-load astragaloside compositions.
Summary of the invention
The objective of the invention is to overcome the deficiency of traditional handicraft, saved the n-butyl alcohol organic solvent, avoided the chromatographic column gradient elution, a kind of efficient, simple method is provided and has obtained high-load astragaloside compositions.
Another object of the present invention is to provide a kind of high-load astragaloside preparation of compositions method by means such as extraction, separation, hydrolysis conversions.
High-load astragaloside compositions reduces impurity, and it is big not only to have improved the dosage that Chinese patent medicine often has, easy moisture absorption, and also in treatment cardio-cerebrovascular diseases drug effect remarkable, the side effect of avoiding some impurity to cause.
The rough Radix Astragali total saponins that obtains from the Radix Astragali comprises acetyl astragaloside, different astragaloside, astragaloside etc., and their basic structure general formula is shown in following (I) formula, and the structural formula of astragaloside is shown in following (II) formula.
In the formula: R
1, R
2, R
3Be Ac or H
All R
1, R
2, R
3The position has A
CThe chemical compound of base, alkali can both make its ester linkage breaking slough acyl group, so the chemical compound of formula (I), can obtain the astragaloside of structural formula for (II) through basic hydrolysis.
Therefore, the technical solution used in the present invention is:
Get rough Radix Astragali total saponins, with the aqueous alkali dissolving fully, through temperature 60-100 ℃ heating in water bath 0.5-3 hour, make its hydrolysis, as seen white precipitate is separated out, the natural cooling after-filtration, and drying gets white precipitate, be astragaloside content greater than 50%, especially greater than 75% astragaloside compositions.In order further to make high-load astragaloside compositions, with above-mentioned white precipitate with the dissolving of methanol reflux after, natural cooling or add the water recrystallization obtains astragaloside greater than 90% compositions.
According to the embodiment of comparative optimization of the present invention, the method for extracting astragalus root methyl-glycoside composition comprises the steps: that (1) get Radix Astragali raw material, uses water as the solvent heating and refluxing extraction; (2) directly go up macroporous resin column behind the extracting liquid filtering, after upper prop was finished, water was rinsed well, used the ethanol elution of 70-95% instead, did not have Radix Astragali total saponins in the eluent (usually in astragaloside) to detecting; (3) eluent is evaporated to dried cream at 60 ℃-75 ℃, gets rough Radix Astragali total saponins; (4) rough Radix Astragali total saponins is fully dissolved with aqueous alkali, through temperature 60-100 ℃ heating in water bath 0.5-3 hour, make its hydrolysis, visible white precipitate is separated out, the natural cooling after-filtration, and drying gets white precipitate.
In above-mentioned (1) step, extract solvent and can use methanol, ethanol or n-butyl alcohol instead, consider that from the production cost angle preferentially use water as solvent to extract, extracting method can be percolation, backflow or other feasible modes of operation.Complete for what guarantee to extract, the consumption of water can suitably increase, and the 10-20 water doubly with raw material weight carries out the hot reflux extraction usually.
For helping the separation of effective ingredient, aqueous extract need be dispersed on the appropriate carriers, be to use HPD-100,300,400,500,600 in the technical scheme for example of the present invention, or D101,1300, AB-8 macroporous resin (not getting rid of other feasible carrier mass), consider preferred HPD-100 macroporous resin from applied sample amount, adsorption rate, desorption rate.After upper prop is finished, clean with flushing with clean water, use ethanol elution then.The optimized technical scheme according to the present invention, with the macroporous resin dress post of handling well in advance, specification and material to used chromatographic column do not have specific (special) requirements, can be selected according to demand of practical production, and operational approach filling routinely, dress post amount be no less than usually post high 2/3, preferably select for use diameter and post height ratio 1: 5-1: 6 post.Alcoholic acid elution speed is preferably the 1000-1500 ml/min.
The eluent of collecting at decompression recycling ethanol below 75 ℃ to dried cream, preferred 65-70 ℃.Get dry extract is further made with extra care through the Saponin basic hydrolysis and is obtained described astragalus root methyl-glycoside composition.The used alkali of the present invention is sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, ammonia, considers from the cost angle, preferred sodium hydroxide.
According to technical scheme of the present invention, with the pharmaceutical preparation that this pharmaceutical composition is made a definite form, said preparation also has solid or liquid excipient except that active component.It is characterized in that: the tablet, capsule, drop pill, soft capsule, powder pin or the large and small injection that contain above-mentioned astragaloside.
The present invention by scientific and feasible method from the Radix Astragali through series of steps such as extraction, separation, hydrolysis conversions, particularly basic hydrolysis transforms this committed step, make astragaloside content greater than 50%, especially greater than 75% astragaloside compositions, meet the raw material standard of Chinese medicine two classes of People's Republic of China's " law of medicine management " regulation, and further said composition and pharmaceutically acceptable carrier are mixed and made into various available preparations.
The invention provides a kind of content height, definite ingredients, quality controllable, drug effect is definite, side effect is little astragaloside composition and method of making the same, said composition is obvious in effects such as treatment chronic nephritis proteinuria, diabetes, cardiovascular and cerebrovascular disease.Preparation technology of the present invention produces checking by many batches, proves that its repeatability, stability are all good, and the yield height of raw material is fit to suitability for industrialized production.
The specific embodiment
Among the embodiment, used extraction water is a tap water, and used ethanol is pharmaceutical grade, and sodium hydroxide, potassium hydroxide, potassium carbonate are technical grade.The Milkvetch Root place of production is Yantai, Shandong; Macroporous resin is available from the precious grace chemical industry company limited in Cangzhou, Hebei and Chemical Plant of Nankai Univ.; The detection of Radix Astragali total saponins is normally in astragaloside, and the method that thin layer chromatography detects astragaloside is being known in the art, and does not limit at this, will not describe in detail.
The assay of astragaloside adopts high performance liquid chromatography (HPLC) external standard method in the Radix Astragali total saponins, and liquid-phase condition is:
Mobile phase: second cyanogen: water=36: 64
Detector: evaporative light scattering detector
Flow velocity: 0.8L/ minute
Chromatographic column: C18 post, 250cm * 4.6mm, particle diameter 5 μ m
Column temperature: 30 ℃
1, standard curve
Accurately take by weighing astragaloside standard substance 10.0mg, be transferred to the 10mL volumetric flask and be settled to scale with dissolve with methanol, therefrom precision measures 0.6,0.8,1.0,1.4,1.8,2.5mL, place respectively the 10mL volumetric flask with methanol constant volume to scale, the respectively therefrom accurate 20 μ L of absorption inject high performance liquid chromatograph, make spectrogram, with concentration is abscissa, and peak area is a vertical coordinate, but the drawing standard curve:
The range of linearity: 60-250 μ g/mL
Regression equation: Y=1.0019+2.0206X
Correlation coefficient: r=0.9991
2, the mensuration of astragaloside in the Radix Astragali total saponins
Accurately take by weighing Radix Astragali total saponins 20.0mg, be transferred to the 100mL volumetric flask and be settled to scale with dissolve with methanol, as need testing solution.The therefrom accurate 20 μ L of absorption inject high performance liquid chromatograph, obtain the content that peak area substitution regression equation is calculated astragaloside.
Embodiment one:
Get the thin slice that exsiccant Radix Astragali raw material is cut into 1-3mm, the hot reflux extraction pot of packing into, with the water logging bubble 1h of 10 times of amounts, heating and refluxing extraction 4h emits extracting solution then, filters, and collects filtrate.
Get the HPD-100 macroporous resin of 1/2 medical material amount, water wet method dress post, alcohol flushing to eluent add water and do not produce muddy ending, and use water then instead and wash residual to there not being ethanol.Above-mentioned filtrate is passed through macroporous resin column, after upper prop is finished, water is rinsed well, use 75% ethanol elution after a while instead, elution speed is 1000 ml/min, collects eluent, at any time detect, to eluent, do not contain Radix Astragali total saponins composition (usually in astragaloside), eluent is evaporated under 70 ℃ dried, the rough Radix Astragali total saponins of pale brown color.
With the dissolving of 1% sodium hydroxide solution, the heating in water bath 3h through 60 ℃ of temperature makes its hydrolysis with above-mentioned rough Radix Astragali total saponins, as seen white precipitate is separated out, the natural cooling after-filtration, through 80 ℃ of dry white precipitates that get, the content that high performance liquid chromatogram is measured astragaloside in this white precipitate is 80%.The methanol hot reflux that this white precipitate is added 30 times of amounts is dissolved, and takes advantage of heat filtering, and the filtrate natural cooling is separated out white needle, and drying is measured the wherein content 95% of astragaloside.
Embodiment two:
Get the thin slice that exsiccant Radix Astragali crude drug is cut into 1-3mm, the hot reflux extraction pot of packing into, with the water logging bubble 1h of 15 times of amounts, heating and refluxing extraction 4h emits extracting solution then, filters, and collects filtrate.
Get the HPD-300 macroporous resin of 1/2 medical material amount, ethanol wet method dress post, alcohol flushing to eluent add water and do not produce muddy ending, and use water then instead and wash residual to there not being ethanol.Above-mentioned filtrate is passed through macroporous resin column, after upper prop is finished, water is rinsed well, use 80% ethanol elution after a while instead, elution speed is 1200 ml/min, collects eluent, at any time detect, to eluent, do not contain astragaloside composition (usually in astragaloside), eluent is evaporated under 65 ℃ dried, the rough Radix Astragali total saponins of pale brown color.
With the dissolving of 3% sodium hydroxide solution, the heating in water bath 2h through 70 ℃ of temperature makes its hydrolysis with above-mentioned rough Radix Astragali total saponins, as seen white precipitate is separated out, the natural cooling after-filtration, through 80 ℃ of dry white precipitates that get, the content that high performance liquid chromatogram is measured astragaloside in this white precipitate is 75%.
Embodiment three:
Get the thin slice that exsiccant Radix Astragali crude drug is cut into 1-3mm, the hot reflux extraction pot of packing into, with the water logging bubble 1h of 15 times of amounts, heating and refluxing extraction 4h emits extracting solution then, filters, and collects filtrate.
Get the HPD-400 macroporous resin of 2/3 medical material amount, ethanol wet method dress post, alcohol flushing to eluent add water and do not produce muddy ending, and use water then instead and wash residual to there not being ethanol.Above-mentioned filtrate is passed through macroporous resin column, after upper prop is finished, water is rinsed well, use 90% ethanol elution after a while instead, elution speed is 1500 ml/min, collects eluent, at any time detect, to eluent, do not contain astragaloside composition (usually in astragaloside), eluent is evaporated under 75 ℃ dried, the rough Radix Astragali total saponins of pale brown color.
With the dissolving of 5% sodium hydroxide solution, the heating in water bath 2h through 80 ℃ of temperature makes its hydrolysis with above-mentioned rough Radix Astragali total saponins, as seen white precipitate is separated out, the natural cooling after-filtration, through 80 ℃ of dry white precipitates that get, the content that high performance liquid chromatogram is measured astragaloside in this white precipitate is 76%.
Embodiment four:
Get the thin slice that exsiccant Radix Astragali crude drug is cut into 1-3mm, the hot reflux extraction pot of packing into, with the water logging bubble 1h of 20 times of amounts, heating and refluxing extraction 4h emits extracting solution then, filters, and collects filtrate.
The AB-8 macroporous resin of the material amount of getting it filled, ethanol wet method dress post, alcohol flushing to eluent add water and do not produce muddy ending, and use water then instead and wash residual to there not being ethanol.Above-mentioned filtrate is passed through macroporous resin column, after upper prop is finished, water is rinsed well, use 95% ethanol elution after a while instead, elution speed is 1200 ml/min, collects eluent, at any time detect, to eluent, do not contain astragaloside composition (usually in astragaloside), eluent is evaporated under 75 ℃ dried, the rough Radix Astragali total saponins of pale brown color.
With the dissolving of 7% sodium hydroxide solution, the heating in water bath 1.5h through 90 ℃ of temperature makes its hydrolysis with above-mentioned rough Radix Astragali total saponins, as seen white precipitate is separated out, the natural cooling after-filtration, through 80 ℃ of dry white precipitates that get, the content that high performance liquid chromatogram is measured astragaloside in this white precipitate is 77%.
Embodiment five:
Get the thin slice that exsiccant Radix Astragali crude drug is cut into 1-3mm, the hot reflux extraction pot of packing into, with the water logging bubble 1h of 15 times of amounts, heating and refluxing extraction 4h emits extracting solution then, filters, and collects filtrate.
The D-101 macroporous resin of the material amount of getting it filled, ethanol wet method dress post, alcohol flushing to eluent add water and do not produce muddy ending, and use water then instead and wash residual to there not being ethanol.Above-mentioned filtrate is passed through macroporous resin column, after upper prop is finished, water is rinsed well, use 85% ethanol elution after a while instead, elution speed is 1200 ml/min, collects eluent, at any time detect, to eluent, do not contain astragaloside composition (usually in astragaloside), eluent is evaporated under 70 ℃ dried, the rough Radix Astragali total saponins of pale brown color.
With the dissolving of 3% potassium hydroxide solution, the heating in water bath 1h through 100 ℃ of temperature makes its hydrolysis with above-mentioned rough Radix Astragali total saponins, as seen white precipitate is separated out, the natural cooling after-filtration, through 80 ℃ of dry white precipitates that get, the content that high performance liquid chromatogram is measured astragaloside in this white precipitate is 75%.
Embodiment six:
Get the thin slice that exsiccant Radix Astragali crude drug is cut into 1-3mm, the hot reflux extraction pot of packing into, with the water logging bubble 1h of 15 times of amounts, heating and refluxing extraction 4h emits extracting solution then, filters, and collects filtrate.
1300 macroporous resins of the material amount of getting it filled, ethanol wet method dress post, alcohol flushing to eluent add water and do not produce muddy ending, and use water then instead and wash residual to there not being ethanol.Above-mentioned filtrate is passed through macroporous resin column, after upper prop is finished, water is rinsed well, use 90% ethanol elution after a while instead, elution speed is 1200 ml/min, collects eluent, at any time detect, to eluent, do not contain astragaloside composition (usually in astragaloside), eluent is evaporated under 60 ℃ dried, the rough Radix Astragali total saponins of pale brown color.
With the dissolving of 5% solution of potassium carbonate, the heating in water bath 1h through 100 ℃ of temperature makes its hydrolysis with above-mentioned rough Radix Astragali total saponins, as seen white precipitate is separated out, the natural cooling after-filtration, through 80 ℃ of dry white precipitates that get, the content that high performance liquid chromatogram is measured astragaloside in this white precipitate is 75%.
Embodiment seven:
Astragaloside compositions 10g
Sodium carboxymethyl cellulose 5g
PEG6000 80g
Sodium lauryl sulphate 5g
Above adjuvant is pulverized mixing in 150 ℃ of heat fused, make it to become molten condition, drip then and make drop pill.
Embodiment eight:
Get astragaloside compositions 1000mg and be dissolved in the 20mL glacial acetic acid, add water and other additives then, through the decarburization filtration sterilization, with the 250mL transfusion that aseptic manipulation is made 5mg/100mL, cryopreservation (less than 15 ℃).
Claims (10)
1, a kind of astragalus root methyl-glycoside composition mainly contains from the Chinese medicine astragalus dry root and forms through extraction, separation, purified astragaloside, and it is characterized in that: in the said composition, the content of astragaloside is more than 50%, especially greater than 75%.
2, according to the described astragalus root methyl-glycoside composition of claim 1, it is characterized in that: the tablet, capsule, drop pill, soft capsule, powder pin or the large and small injection that contain above-mentioned astragaloside.
3, according to the described astragalus root methyl-glycoside composition of claim 1, the application in preparation treatment chronic nephritis proteinuria, diabetes, cardiovascular and cerebrovascular vessel medicine.
4, a kind of method for preparing astragalus root methyl-glycoside composition from the Radix Astragali is a raw material with the Radix Astragali, uses water extraction, and prepares the astragaloside compositions through separation, purification, hydrolysis, it is characterized in that comprising the steps:
(1) gets Radix Astragali raw material, use water as the solvent heating and refluxing extraction;
(2) directly go up macroporous resin column behind the extracting liquid filtering, after upper prop was finished, water was rinsed well, used the ethanol elution of 70-95% instead, did not have Radix Astragali total saponins in the eluent (usually in astragaloside) to detecting;
(3) eluent is evaporated to dried cream at 60 ℃-75 ℃, gets rough Radix Astragali total saponins;
(4) rough Radix Astragali total saponins is fully dissolved with aqueous alkali, through temperature 60-100 ℃ heating in water bath 0.5-3 hour, make its hydrolysis, visible white precipitate is separated out, the natural cooling after-filtration, and drying gets white precipitate.
5, method according to claim 4 is characterized in that used macroporous resin is HPD-100,300,400,500,600, or D101,1300, AB-8.
6, method according to claim 4 is characterized in that alcoholic acid elution speed is the 1000-1500 ml/min.
7, method according to claim 4 is characterized in that used alkali is industrial bases, as sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, ammonia.
8, method according to claim 4, it is characterized in that it comprises that also the white depositions that will obtain is through further purified process, described further making with extra care comprises: after white precipitate is dissolved with the methanol reflux, natural cooling or add the water recrystallization obtains astragaloside content in the compositions more than 90%.
9, according to the described astragaloside preparation method of composition of one of claim 4 to 7, it is characterized in that: in the described astragaloside compositions, the content of astragaloside is more than 50%, especially greater than 75%.
10, according to the described astragaloside preparation method of composition of one of claim 4 to 7, it is characterized in that: described astragaloside compositions, can mix with pharmaceutically acceptable carrier.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005084689A1 (en) * | 2004-02-23 | 2005-09-15 | Hutchison Medipharma Enterprises Limited | Radix astragli extract and production method and use thereof |
| CN100333731C (en) * | 2005-09-08 | 2007-08-29 | 南京医科大学 | Application of astragaloside in preparing medication for treating peripheral nervous lesion of diabetes |
| CN1569884B (en) * | 2004-04-29 | 2010-11-10 | 南京医科大学 | Method for preparing astragaloside and its use in preparation of drug for preventing and treating diabetic nephropathy |
| CN1939330B (en) * | 2005-09-13 | 2010-11-10 | 山东绿叶天然药物研究开发有限公司 | Medical usage of milkvetch-root methcoside |
| CN1854148B (en) * | 2005-02-06 | 2011-05-11 | 成都地奥九泓制药厂 | Astragalus total heteroside extract and its production |
| EP2548880A3 (en) * | 2003-06-23 | 2013-06-19 | Geron Corporation | Compositions for increasing telomerase activity |
| CN107365344A (en) * | 2017-07-21 | 2017-11-21 | 安徽龙津生物科技有限公司 | A kind of method of the extraction purification Astragaloside IV from astragalus root |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101225424B (en) | 2007-09-13 | 2013-05-29 | 天津药物研究院 | Single-glucopyranoside of cyclomembranousol, preparation method, medicament combination and uses thereof |
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2002
- 2002-07-22 CN CN 02137982 patent/CN1189176C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2548880A3 (en) * | 2003-06-23 | 2013-06-19 | Geron Corporation | Compositions for increasing telomerase activity |
| WO2005084689A1 (en) * | 2004-02-23 | 2005-09-15 | Hutchison Medipharma Enterprises Limited | Radix astragli extract and production method and use thereof |
| CN1569884B (en) * | 2004-04-29 | 2010-11-10 | 南京医科大学 | Method for preparing astragaloside and its use in preparation of drug for preventing and treating diabetic nephropathy |
| CN1854148B (en) * | 2005-02-06 | 2011-05-11 | 成都地奥九泓制药厂 | Astragalus total heteroside extract and its production |
| CN100333731C (en) * | 2005-09-08 | 2007-08-29 | 南京医科大学 | Application of astragaloside in preparing medication for treating peripheral nervous lesion of diabetes |
| CN1939330B (en) * | 2005-09-13 | 2010-11-10 | 山东绿叶天然药物研究开发有限公司 | Medical usage of milkvetch-root methcoside |
| CN107365344A (en) * | 2017-07-21 | 2017-11-21 | 安徽龙津生物科技有限公司 | A kind of method of the extraction purification Astragaloside IV from astragalus root |
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| CN1189176C (en) | 2005-02-16 |
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Effective date of registration: 20160322 Address after: 223001 No. 9 HANKOOK North Road, Jiangsu, Huaian Patentee after: Jiangsu Chiatai Qingjiang Pharmaceutical Co., Ltd. Address before: 222006 Sinpo City, Lianyungang Province, North Road, No. 8, No. Patentee before: Chia Tai Tianqing Pharmaceutical Group Co., Ltd. |
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Granted publication date: 20050216 Termination date: 20170722 |