CN102772462A - Method for extracting and separating total saponins of panax ginseng from american ginseng - Google Patents
Method for extracting and separating total saponins of panax ginseng from american ginseng Download PDFInfo
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Abstract
The invention discloses a method for extracting and separating total saponins of panax ginseng from american ginseng. The method employs a series of high-efficiency extraction, separation and purification technical measures of alcohol-backflow extracting, water precipitating, purifying by a macroporous adsorption resin, decolorizing with an ion exchange resin, etc. The obtained total saponins of panax ginseng are white powder and comprise ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd, wherein the total purity of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 is more than 74.0%. The production process is simple and practical, has no pollution, and is suitable for large-scale production.
Description
Technical field
The present invention relates to a kind of from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins.
Background technology
Radix Panacis Quinquefolii (Panax quinquefolium L.) is Araliaceae (Araliaceae) Panax (Panax) herbaceos perennial, originates in eastern united states and Canada.Have another name called U.S.'s Radix Ginseng, Radix Panacis Quinquefolii, Radix Panacis Quinquefolii and Guangzhou ginseng.Beginning is stated from supplementary Amplifications of the Compendium of Materia Medica in China, the medicinal history in existing more than 200 year.Have effects such as lung benefiting the moon, clear asthenic fire, promoting the production of body fluid to quench thirst, control chronic cough of deficiency lung, lose blood, dry throat and mouth, deficiency-heat be tired tired etc." Records of Tradition Chinese and Western Medicine in Combination " carries: " Radix Panacis Quinquefolii, cool in nature and mend, all desires do not receive the temperature compensation person of the Radix Ginseng with Radix Ginseng, all can this generation it." just because of Radix Panacis Quinquefolii has the benefit property of Radix Ginseng, and there is not the dry property of Radix Ginseng, be described as famous and precious tonic and well-known.
Contain saponins, polyacetylene class, volatile oil, polysaccharide and amino acids in the Radix Panacis Quinquefolii, wherein main active component is a saponins compound.Big quantity research shows that because the difference of structure, various ginsenosides have its unique pharmacologically active; Thereby pharmacologically active is multiformity: the ginsenoside has resisting fatigue; Slow down aging is regulated the central nervous system, human body immunity improving power; Improve the cardiovascular and cerebrovascular vessel blood supply insufficiency, suppress effects such as growth of tumour cell.
The extraction of some American ginseng total saponins, the patent of invention of purification are disclosed in recent years.One Chinese patent application 03127613.X discloses a kind of method of from Fructus Panacis Quinquefolii, extracting, making with extra care Radix Ginseng total saponins; With Fructus Panacis Quinquefolii carry out that water is carried, macroporous adsorbent resin remove impurity, decolouring, drying; Though the method is simple, color is dark, yield and purity are not high.One Chinese patent application number 03127632.6 discloses a kind of production technology from Radix Panacis Quinquefolii and the refining total saponins of stem and leaf of Radix Panacis Quinquefolii; Radix Panacis Quinquefolii is crossed stem and leaf of Radix Panacis Quinquefolii pulverize and to be coarse powder, water is carried, precipitate with ethanol, alkali deposited, macroporous adsorbent resin remove impurity, activated carbon decolorizing, lyophilizing.The purity of the method gained total saponins is higher, but complex steps, yield are low.One Chinese patent application numbers 200610014229.6 discloses a kind of method for preparing of American ginseng total saponins, and Radix Panacis Quinquefolii is added solvent, extract after transferring alkali, and the remove impurity of macroporous type anion exchange resin, decolouring, the method possibly make some ginsenoside's structure change.One Chinese patent application numbers 200610109414.3 discloses a kind of method for preparing of from Radix Panacis Quinquefolii or stem and leaf of Radix Panacis Quinquefolii, extracting total saponins; Circulated in countercurrent supersound extraction, precipitate with ethanol, macroporous resin column remove impurity, decolorizing column decolouring, spray drying etc., the method step is more loaded down with trivial details.
Summary of the invention
The present invention provide a kind of from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins, this method comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 6-12 at every turn and doubly measure the 30%-70% alcoholic solution, heating and refluxing extraction 2-3 time, each 1h-3h soaks 60min-90min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:2-2:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 3 ~ 5 times, shake up, place refrigerator 12h ~ 24h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:2-1.6:1 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:3-1:13, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 4.5-9 times of column volume/hour, saturated 1h-3h; Carry out eluting with the distilled water of 5 times of-10 times of column volumes and remove sugar, elution flow rate be 4.5-12 times of column volume/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 4.5-12 times of column volume/hour; Alcoholic solution with 5-15 times of column volume 70% carries out eluting, elution flow rate be 4.5-12 times of column volume/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 3:1-1:1 takes by weighing weak-base anion-exchange resin; The blade diameter length ratio of packing into is the chromatographic column of 1:3-1:6; Use the acid-alkali treatment weak-base anion-exchange resin, finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 3 times of-5 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The present invention's method of extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii is preferably and comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 6 times of amount 70% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 3h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:2, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 5 times, shake up, place refrigerator 12h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:2 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:13, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 4.5 times of column volumes/hour, saturated 3h; Carry out eluting with the distilled water of 5 times of column volumes and remove sugar, elution flow rate be 12 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 4.5 times of column volumes/hour; Alcoholic solution with 15 times of column volumes 70% carries out eluting, elution flow rate be 4.5 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 1:1 takes by weighing weak-base anion-exchange resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:3, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 5 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The method of the present invention's extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii also is preferably and comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 12 times of amount 30% alcoholic solution at every turn, heating and refluxing extraction 2 times, each 1h soaks 90min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 2:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 3 times, shake up, place refrigerator 24h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1.6:1 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:3, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 9 times of column volumes/hour, saturated 1h; Carry out eluting with the distilled water of 10 times of column volumes and remove sugar, elution flow rate be 4.5 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 12 times of column volumes/hour; Alcoholic solution with 5 times of column volumes 70% carries out eluting, elution flow rate be 12 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 2:1 takes by weighing weak-base anion-exchange resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:6, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 3 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The method of the present invention's extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii also is preferably and comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 9 times of amount 50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 2h soaks 75min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 4 times, shake up, place refrigerator 18h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:1.5 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:8, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 6 times of column volumes/hour, saturated 2h; Carry out eluting with the distilled water of 7.5 times of column volumes and remove sugar, elution flow rate be 6 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 6 times of column volumes/hour; Alcoholic solution with 10 times of column volumes 70% carries out eluting, elution flow rate be 6 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 3:2 takes by weighing weak-base anion-exchange resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:4.5, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 4 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
Employed macroporous adsorbent resin is preferably a kind of in HPD100, HPD100A, HPD300, HPD450, AB-8 or the D101 resin in the method for the present invention's extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii, and the weak-base ion-exchange resin that uses is 330 resins.
The method of the present invention's extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii further is preferably and comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 10 times of amount 50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 1h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 5 times, shake up, place refrigerator 12h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:1 takes by weighing the AB-8 macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:6, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 4.5 times of column volumes/hour, saturated 1h; Carry out eluting with the distilled water of 10 times of column volumes and remove sugar, elution flow rate be 12 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 12 times of column volumes/hour; Alcoholic solution with 5 times of column volumes 70% carries out eluting, elution flow rate be 12 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 3:1 takes by weighing 330 weak-base anion-exchange resins, and the blade diameter length ratio of packing into is the chromatographic column of 1:6, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 3 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The Latin formal name used at school of the raw materials of traditional Chinese medicinal materials Radix Panacis Quinquefolii in the method for the present invention's extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii: PANACIS QUINQUEFOLII RADIX is the dry root of Araliaceae Radix Panacis Quinquefolii Panax quique folium L..
Radix Panacis Quinquefolii coarse powder of the present invention is meant " coarse powder " stipulated in the Chinese Pharmacopoeia note on the use, that is: referring to can be all through No. two sieve.Can be no more than 40% powder through No. four sieves but be mixed with.The volume of the solvent in the leaching process of the present invention is to calculate according to the weight of medical material; As use 70% ethanol of 6 times of amounts; Be meant the per kilogram medical material, add following 6 liter 70% ethanol of room temperature, i.e. w/v; The envelope-bulk to weight ratio of spissated medicinal liquid and medical material also is meant the per kilogram medical material, and what rise, such as: " be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:2 " is that the per kilogram medical material is concentrated into 2L.
Adopt the method for the present invention's extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii; Continuous parameters according to embodiment 1 is produced five batches of Radix Ginseng total saponinss; HPLC analyzing total saponin comprises ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, Ginsenoside Rc, ginsenoside Rd, and wherein total purity of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 is more than 74.0%.Detailed results is seen table 1
Continuous five crowdes of purity and the rate of transform results that produce astragaloside of table 1
| Lot number | Ginsenoside Rg1's content % | Ginsenoside Re's content % | Ginsenoside Rb1's content % | Rate of transform % |
| 071101 | 2.22 | 19.02 | 53.95 | 100.0 |
| 071112 | 2.12 | 19.29 | 52.69 | 99.1 |
| 071124 | 2.26 | 19.27 | 53.09 | 100.0 |
| 071203 | 2.24 | 19.14 | 53.73 | 99.8 |
| 071219 | 2.28 | 19.20 | 53.28 | 99.4 |
This shows, process for purification of the present invention, process stabilizing is reliable, is easy to suitability for industrialized production, and total purity of the ginsenoside Rg1 who produces, ginsenoside Re and ginsenoside Rb1 is more than 74.0%.And have the following advantages: this production technology is simple, and is pollution-free, is suitable for big production; And the purity of resulting Radix Ginseng total saponins and yield are all higher.
The specific embodiment
Following embodiment is used for illustrating the method for the present invention from Radix Panacis Quinquefolii extraction separation Radix Ginseng total saponins, but it can not constitute any restriction to scope of the present invention.
Embodiment 1:
A, extraction: take by weighing Radix Panacis Quinquefolii coarse powder 10kg, add the 60L70% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 3h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and standardize solution 20L gets concentrated solution;
B, water precipitating: the concentrated solution thin up to 100L, is shaken up, place refrigerator 12h, filter, discard filtering residue, get extracting solution;
C, purification for the first time: take by weighing HPD100 macroporous resin 6.25kg, the diameter of packing into is 9.4cm, highly be 122cm, and volume is the chromatographic column of 8.5L, with 95% ethanol and distilled water processing totally; With appearance on the extracting solution, the absorption flow velocity is 38L/h, saturated 3h; Distilled water with 42.5L carries out eluting except that sugar, and elution flow rate is 102L/h; With the remove impurity of 60L20% alcoholic solution, elution flow rate is 38L/h; Alcoholic solution with 127L70% carries out eluting, and elution flow rate is 38L/h, collects 70% ethanol elution, gets refined solution A;
D, purification for the second time: take by weighing 330 weak-base anion-exchange resin 10kg, the diameter of packing into is 18.2cm, highly is 54.6cm, and volume is the chromatographic column of 14.2L, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, and flow velocity is 71L, collects effluent; Flow through the weak anion resin post with 71L 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The result: ginsenoside Rg1's content is 2.15%, ginsenoside Re's content be 19.03% and ginsenoside Rb1's content be 53.77%, changeing interest rate is 99.6%.
Embodiment 2:
A, extraction: take by weighing Radix Panacis Quinquefolii coarse powder 15Kg, add the 180L30% alcoholic solution at every turn, heating and refluxing extraction 2 times, each 1h soaks 90min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, is settled to 30L, gets concentrated solution;
B, water precipitating: the concentrated solution thin up to 90L, is shaken up, place refrigerator 24h, filter, discard filtering residue, get extracting solution;
C, purification for the first time: take by weighing HPD100A macroporous adsorbent resin 15kg, the diameter of packing into is 20cm, highly be 60cm, and volume is the chromatographic column of 19L, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 9 times of column volumes of 171L/h/hour, saturated 1h; Distilled water with 190L carries out eluting except that sugar, and elution flow rate is 86L/h; With the remove impurity of 133L20% alcoholic solution, elution flow rate is 228L/h; Alcoholic solution with 190L70% carries out eluting, and elution flow rate is 95L/h, collects 70% ethanol elution, gets refined solution A;
D, purification for the second time: take by weighing D900 weak-base anion-exchange resin 7.5kg, the diameter of packing into is 13cm, highly is 80cm, and volume is the chromatographic column of 11L, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, and flow velocity is 55L/h, collects effluent; Flow through the weak anion resin post with the 33L70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The result: ginsenoside Rg1's content is 2.20%, ginsenoside Re's content be 19.25% and ginsenoside Rb1's content be 53.12%, changeing interest rate is 99.7%.
Embodiment 3:
A, extraction: take by weighing Radix Panacis Quinquefolii coarse powder 20kg, add the 180L50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 2h soaks 75min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, is settled to 20L, gets concentrated solution;
B, water precipitating: the concentrated solution thin up to 80L, is shaken up, place refrigerator 18h, filter, discard filtering residue, get extracting solution;
C, purification for the first time: take by weighing HPD300 macroporous adsorbent resin 13Kg, the diameter of packing into is 14cm, highly be 112cm, and volume is the chromatographic column of 17L, with 95% ethanol and distilled water processing totally; With appearance on the extracting solution, the absorption flow velocity is 102L/h, saturated 2h; Distilled water with 127.5L carries out eluting except that sugar, and elution flow rate is 102L/h; With the remove impurity of 119L20% alcoholic solution, elution flow rate is 102L/h; Alcoholic solution with 170L70% carries out eluting, and elution flow rate is 102L/h, collects 70% ethanol elution, gets refined solution A;
D, purification for the second time: take by weighing 335 weak-base anion-exchange resin 13kg, the diameter of packing into is 17.5cm, highly is 80cm, and volume is the chromatographic column of 19L, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, and flow velocity is 95L/h, collects effluent; Flow through the weak anion resin post with the 76L70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The result: ginsenoside Rg1's content is 2.17%, ginsenoside Re's content be 19.23% and ginsenoside Rb1's content be 53.06%, changeing interest rate is 99.0%.
Embodiment 4:
A, extraction: take by weighing Radix Panacis Quinquefolii coarse powder 10kg, add the 100L50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 1h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, is settled to 10L, gets concentrated solution;
B, water precipitating: the concentrated solution thin up to 50L, is shaken up, place refrigerator 12h, filter, discard filtering residue, get extracting solution;
C, purification for the first time: take by weighing AB-8 macroporous adsorbent resin 10kg, the diameter of packing into is 15cm, highly be 85cm, and volume is the chromatographic column of 15L, with 95% ethanol and distilled water processing totally; With appearance on the extracting solution, the absorption flow velocity is 67.5L/h, saturated 1h; Distilled water with 150L carries out eluting except that sugar, and elution flow rate is 180L/h; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate is 180L/h; Alcoholic solution with 75L70% carries out eluting, and elution flow rate is 180L/h, collects 70% ethanol elution, gets refined solution A;
D, purification for the second time: take by weighing 330 weak-base anion-exchange resin 3.3kg, the diameter of packing into is 10cm, highly is 65cm, and volume is the chromatographic column of 5L, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, and flow velocity is 25L/h, collects effluent; Flow through the weak anion resin post with the 15L70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The result: ginsenoside Rg1's content is 2.29%, ginsenoside Re's content be 19.32% and ginsenoside Rb1's content be 53.54%, changeing interest rate is 100.0%.
Embodiment 5:
A, extraction: take by weighing Radix Panacis Quinquefolii coarse powder 10kg, add the 60L70% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 3h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and standardize solution 20L gets concentrated solution;
B, water precipitating: the concentrated solution thin up to 100L, is shaken up, place refrigerator 12h, filter, discard filtering residue, get extracting solution;
C, purification for the first time: take by weighing HPD450 macroporous resin 6.25kg, the diameter of packing into is 9.4cm, highly be 122cm, and volume is the chromatographic column of 8.5L, with 95% ethanol and distilled water processing totally; With appearance on the extracting solution, the absorption flow velocity is 38L/h, saturated 3h; Distilled water with 42.5L carries out eluting except that sugar, and elution flow rate is 102L/h; With the remove impurity of 60L20% alcoholic solution, elution flow rate is 38L/h; Alcoholic solution with 127L70% carries out eluting, and elution flow rate is 38L/h, collects 70% ethanol elution, gets refined solution A;
D, purification for the second time: take by weighing D900 weak-base anion-exchange resin 10kg, the diameter of packing into is 18.2cm, highly is 54.6cm, and volume is the chromatographic column of 14.2L, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, and flow velocity is 71L, collects effluent; Flow through the weak anion resin post with 71L 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The result: ginsenoside Rg1's content is 2.37%, ginsenoside Re's content be 18.81% and ginsenoside Rb1's content be 52.91%, changeing interest rate is 99.87%.
Embodiment 6:
A, extraction: take by weighing Radix Panacis Quinquefolii coarse powder 10kg, add the 100L50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 1h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, is settled to 10L, gets concentrated solution;
B, water precipitating: the concentrated solution thin up to 50L, is shaken up, place refrigerator 12h, filter, discard filtering residue, get extracting solution;
C, purification for the first time: take by weighing D101 macroporous adsorbent resin 10kg, the diameter of packing into is 15cm, highly be 85cm, and volume is the chromatographic column of 15L, with 95% ethanol and distilled water processing totally; With appearance on the extracting solution, the absorption flow velocity is 67.5L/h, saturated 1h; Distilled water with 150L carries out eluting except that sugar, and elution flow rate is 180L/h; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate is 180L/h; Alcoholic solution with 180L70% carries out eluting, and elution flow rate is 180L/h, collects 70% ethanol elution, gets refined solution A;
D, purification for the second time: take by weighing 330 weak-base anion-exchange resin 3.3kg, the diameter of packing into is 10cm, highly is 65cm, and volume is the chromatographic column of 5L, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, and flow velocity is 25L/h, collects effluent; Flow through the weak anion resin post with the 15L70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
The result: ginsenoside Rg1's content is 2.48%, ginsenoside Re's content be 19.85% and ginsenoside Rb1's content be 54.68%, changeing interest rate is 99.95%.
Claims (6)
1. the method for an extraction separation Radix Ginseng total saponins from Radix Panacis Quinquefolii, this method comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 6-12 at every turn and doubly measure the 30%-70% alcoholic solution, heating and refluxing extraction 2-3 time, each 1h-3h soaks 60min-90min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:2-2:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 3 ~ 5 times, shake up, place refrigerator 12h ~ 24h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:2-1.6:1 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:3-1:13, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 4.5-9 times of column volume/hour, saturated 1h-3h; Carry out eluting with the distilled water of 5 times of-10 times of column volumes and remove sugar, elution flow rate be 4.5-12 times of column volume/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 4.5-12 times of column volume/hour; Alcoholic solution with 5-15 times of column volume 70% carries out eluting, elution flow rate be 4.5-12 times of column volume/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 3:1-1:1 takes by weighing weak-base anion-exchange resin; The blade diameter length ratio of packing into is the chromatographic column of 1:3-1:6; Use the acid-alkali treatment weak-base anion-exchange resin, finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 3 times of-5 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
2. according to claim 1 from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins, it is characterized in that this method comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 6 times of amount 70% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 3h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:2, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 5 times, shake up, place refrigerator 12h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:2 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:13, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 4.5 times of column volumes/hour, saturated 3h; Carry out eluting with the distilled water of 5 times of column volumes and remove sugar, elution flow rate be 12 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 4.5 times of column volumes/hour; Alcoholic solution with 15 times of column volumes 70% carries out eluting, elution flow rate be 4.5 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the amount of resin weight ratio is that 1:1 takes by weighing weak-base anion-exchange resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:3, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 5 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
3. according to claim 1 from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins, it is characterized in that this method comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 12 times of amount 30% alcoholic solution at every turn, heating and refluxing extraction 2 times, each 1h soaks 90min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 2:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 3 times, shake up, place refrigerator 24h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1.6:1 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:3, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 9 times of column volumes/hour, saturated 1h; Carry out eluting with the distilled water of 10 times of column volumes and remove sugar, elution flow rate be 4.5 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 12 times of column volumes/hour; Alcoholic solution with 5 times of column volumes 70% carries out eluting, elution flow rate be 12 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 2:1 takes by weighing weak-base anion-exchange resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:6, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 3 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
4. according to claim 1 from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins, it is characterized in that this method comprises the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 9 times of amount 50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 2h soaks 75min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 4 times, shake up, place refrigerator 18h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:1.5 takes by weighing macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:8, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 6 times of column volumes/hour, saturated 2h; Carry out eluting with the distilled water of 7.5 times of column volumes and remove sugar, elution flow rate be 6 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 6 times of column volumes/hour; Alcoholic solution with 10 times of column volumes 70% carries out eluting, elution flow rate be 6 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 3:2 takes by weighing weak-base anion-exchange resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:4.5, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 4 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
According to claim 1-4 each described from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins; It is characterized in that said macroporous adsorbent resin is a kind of in HPD100, HPD100A, HPD300, HPD450, AB-8 or the D101 resin, said weak-base ion-exchange resin is 330 resins.
6. according to claim 5 from Radix Panacis Quinquefolii the method for extraction separation Radix Ginseng total saponins, it is characterized in that comprising the steps:
A, extraction: take by weighing the Radix Panacis Quinquefolii coarse powder, add 10 times of amount 50% alcoholic solution at every turn, heating and refluxing extraction 3 times, each 1h soaks 60min first, filters; Filtrating merges, and being evaporated to does not have the alcohol flavor, and be settled to medicinal liquid: the medical material envelope-bulk to weight ratio is 1:1, gets concentrated solution;
B, water precipitating: with concentrated solution thin up to 5 times, shake up, place refrigerator 12h, filter, discard filtering residue, extracting solution;
C, purification for the first time: according to the medical material amount: the weight ratio of amount of resin is that 1:1 takes by weighing the AB-8 macroporous adsorbent resin, and the blade diameter length ratio of packing into is the chromatographic column of 1:6, with 95% ethanol and distilled water processing totally; With on the extracting solution appearance, absorption flow velocity be 4.5 times of column volumes/hour, saturated 1h; Carry out eluting with the distilled water of 10 times of column volumes and remove sugar, elution flow rate be 12 times of column volumes/hour; With the 20% alcoholic solution remove impurity of 7 times of column volumes, elution flow rate be 12 times of column volumes/hour; Alcoholic solution with 5 times of column volumes 70% carries out eluting, elution flow rate be 12 times of column volumes/hour, collect 70% ethanol elution, refined solution A;
D, purification for the second time: according to the medical material amount: the weight ratio of amount of resin is that 3:1 takes by weighing 330 weak-base anion-exchange resins, and the blade diameter length ratio of packing into is the chromatographic column of 1:6, uses the acid-alkali treatment weak-base anion-exchange resin, and finally using distilled water flushing to pH value is 7; Refined solution A is flow through the weak-base anion-exchange resin bed, flow velocity be 5 times of column volumes/hour, collect effluent; Flow through the weak anion resin post with 3 times of column volume 70% alcoholic solution, collect effluent; Two-part effluent merging is concentrated into nothing alcohol flavor, gets refined solution B;
E, lyophilizing:, promptly get Radix Ginseng total saponins with refined solution B lyophilizing.
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