CN103554209B - Method for preparing ginsenoside Rg1 from pseudo-ginseng - Google Patents
Method for preparing ginsenoside Rg1 from pseudo-ginseng Download PDFInfo
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- CN103554209B CN103554209B CN201310572043.2A CN201310572043A CN103554209B CN 103554209 B CN103554209 B CN 103554209B CN 201310572043 A CN201310572043 A CN 201310572043A CN 103554209 B CN103554209 B CN 103554209B
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- pseudo
- ginseng
- ginsenoside
- methyl alcohol
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Abstract
The invention provides a method for preparing ginsenoside Rg1 from pseudo-ginseng. The method is characterized by comprising the specific steps of cleaning, slicing, drying and grinding fresh pseudo-ginseng root to obtain pseudo-ginseng powder; heating, refluxing and extracting the pseudo-ginseng powder through an organic solvent, and combining extracting solutions; implementing rotary evaporation to the combined extracting solution and recovering the solvent to obtain a pseudo-ginseng extract; dissolving the pseudo-ginseng extract through an organic solvent, mixing a sample through chromatography silica gel, applying onto a silica gel column through a dry method after the organic solvent completely volatizes, implementing gradient elution through methylene dichloride and methanol, collecting an elution part containing the ginsenoside Rg1, and recovering the solvent to obtain a crude product containing the Rg1; dissolving the crude product containing the Rg1 through acetic ether, cooling to separate out a crystal, and filtering to obtain a Rg1 pure product. The method is simple and convenient in process, simple to operate, low in requirement on apparatus and equipment, and can prepare the Rg1 pure product; and the method, which is few in processing step and high in product yield, is suitable for industrial production.
Description
Technical field
The present invention relates to a kind of novel method preparing ginsenoside Rg1 from pseudo-ginseng, particularly relate to the preparation method of a kind of extraction and isolation and purified ginsenoside Rg1 from pseudo-ginseng, belong to natural active product preparation method technical field.
Background technology
Pseudo-ginseng is the dry root of panax araliaceae plant panax notoginseng (Burk) F.H.chen, is China's rare traditional Chinese medicine, and this product has the hemostasis of the loose stasis of blood, subduing swelling and relieving pain.For spitting of blood, spit blood, bleeding from five sense organs or subcutaneous tissue, has blood in stool, uterine bleeding, traumatic hemorrhage, chest ventral spine pain, treating swelling and pain by traumatic injury.Be mainly used in now the treatment of the heart, cerebrovascular system disease.Complicated component in pseudo-ginseng, nearly hundred kinds of compositions have been separated from his root, leaf, carpopodium, bud, rhizome, mainly contain saponins, volatile oil, amino acids, flavonoid, sterols, carbohydrate, organic acid, CYCLIC DIPEPTIDES, inorganic acids, but its principle active component is arasaponin.
In recent years domestic and international research shows, the biological activity of monomer is most important, because the existence of other compositions can be got rid of, be easy to definite pharmacological action and the analysis mechanisms of illustrating him, be easy to find active stronger compound by the structure activity study of various of monomer saponin(e, in current pseudo-ginseng, monomeric compound focuses mostly in the higher composition of Rg1, Rb1, Re, Ra equal size, now prove that ginsenoside Rg1 is effective for senile dementia treatment, Re and Rg2 anti-arrhythmia is comparatively strong, and antitumor Rh2 and Rg3 is stronger.Different just because of monomer saponin effect, have and need to make monomer saponin preparation.The method finding Extraction and isolation monomer from medicinal material is even more important, and a kind of simple process, method simple to operate are the targets of separating monomer.
Ginsenoside Rg1: molecular formula: C
42h
720
14, molecular weight 801.03,
Structural formula is:
Ginsenoside Rg1 has anti-ageing, anti-oxidant, raising immunizing power, and the functions such as memory, have pharmacological action widely to cardiovascular and cerebrovascular, neural system, immunity system.Ginsenoside Rg1 is panaxitriol structure, is first to find from ginseng, and in ginseng, content is lower, but content is higher in pseudo-ginseng.Separation and purification sterling in plant, one is complex process, and two is that component content is few, needs reasonable and simple and direct technique, purifying compounds can be made to become feasible.About the method preparing ginsenoside Rg1 has multiple, some alcohol reflux, upper silica gel or alumina column are separated, some ultrasonic wave refluxing extraction, and extract acid-base precipitation is separated, then are separated with column chromatography silica gel or alumina column.What also have obtains total saponins with D101 macroporous resin, and recycle silicon glue post is separated and obtains sterling, some patent applied fors, but above technical process is long, complicated operation, obtains sterling Rg1 more difficult.
Summary of the invention
The object of this invention is to provide a kind of simple process, simple to operation, product yield is high, proves process stabilizing, from pseudo-ginseng, effectively prepare the novel method of ginsenoside Rg1 through tens of experiments.
In order to achieve the above object, the invention provides a kind of method preparing ginsenoside Rg1 from pseudo-ginseng, it is characterized in that, concrete steps are:
The first step: fresh Radix Notoginseng is cleaned, cuts into slices, dries, is ground into Radix Notoginseng powder;
Second step: the Radix Notoginseng powder the first step obtained is placed in round-bottomed flask or refluxing extraction device, with organic solvent heating and refluxing extraction at least one times, united extraction liquid;
3rd step: the extracting solution rotary evaporation recycling design obtained by second step, obtains Radix Notoginseng extract;
4th step: the Radix Notoginseng extract organic solvent dissolution that the 3rd step is obtained, sample is mixed with silica gel for chromatography, after organic solvent volatilization is clean, silicagel column in dry method, through methylene dichloride and methanol elution gradient, thin-layer chromatography chromatogram detects wash-out process, collects the elution fraction containing ginsenoside Rg1, after recycling design, obtain the crude product containing Rg1;
5th step: by obtain containing the crude product of Rg1, dissolve with vinyl acetic monomer, when being cooled to 5 DEG C-10 DEG C, crystallize out, filters, obtains Rg1 sterling.
Preferably, the organic solvent in described second step is methyl alcohol, and the volume ratio of Radix Notoginseng powder and organic solvent is 1:2-3, and the time of each reflux is 1-3 hour, and the number of times of heating and refluxing extraction is 4-5 time.
Preferably, the temperature of the rotary evaporation in described 3rd step is 0-50 DEG C, pressure is 0.07-0.1Mpa.
Preferably, organic solvent in described 4th step is methyl alcohol, the weightmeasurement ratio of Radix Notoginseng extract and methyl alcohol is 20g:200ml (w/v), Radix Notoginseng extract is 20g:1200g (w/w) with the weight ratio of mixing sample silica gel, and the methylene dichloride of wash-out and the volume ratio of methyl alcohol are 7:1-1:1.
Preferably, measure content by high performance liquid chromatography (HPLC) in described 5th step, actual conditions is: C18250 × 4.6mm chromatographic column, is that acetonitrile and the water of 19.5:80.5 is moving phase with volume ratio, flow velocity 1.0ml/min, determined wavelength is 210nm.
Preferably, in described 4th step, silica gel used is 200-300 order column chromatography silica gel.
Preferably, the testing conditions of the thin-layer chromatography chromatogram in described 4th step is: adopt GF254 plate, developping agent is the mixed solution of trichloromethane, vinyl acetic monomer, first alcohol and water, the volume ratio of described trichloromethane, vinyl acetic monomer, first alcohol and water is 15:40:22:10, with ginsenoside Rg1's reference substance for contrast, upright ascending development, develop the color with 10% ethanol solution of sulfuric acid spraying or develop the color with iodine vapor, 105 DEG C of heating 10 minutes, violet spot is shown during 10% ethanol solution of sulfuric acid, during iodine vapor colour developing, it is yellow spotting.
In preparation method of the present invention, the yield of ginsenoside Rg1 is 0.8% (in dry medicinal material), and purity is 98%, and the ginsenoside Rg1 of gained can be used for pharmaceutical industries and other purposes.
Compared with prior art, the invention has the beneficial effects as follows:
Present invention process is easy, simple to operate, and instrument and equipment requires low, and one time solvent refluxing extracts, and extract is once gone up silica gel column chromatography and is separated, and can obtain Rg1 sterling; Because processing step is few, product yield is high, is applicable to suitability for industrialized production.
Embodiment
For making the present invention become apparent, hereby with preferred embodiment, be described in detail below.
Embodiment
By fresh Radix Notoginseng 5.5kg, clean, section, dry, put in pulverizer and be ground into Radix Notoginseng powder, about 5kg, above-mentioned Radix Notoginseng powder is placed in refluxing extraction device, adds chemical pure methyl alcohol, the volume ratio of Radix Notoginseng powder and methyl alcohol is 1:2.3, refluxing extraction four times, each extraction 3 hours, each extracting liquid filtering extracting gained, united extraction liquid, 50 DEG C, pressure uses Rotary Evaporators decompression and solvent recovery under being the condition of 0.08Mpa, obtain extract medicinal extract 20g, extraction yield is 0.4%.By extract medicinal extract 200ml dissolve with methanol, sample is mixed with 1200g200-300 order column chromatography silica gel, after heating in water bath makes methyl alcohol volatilization dry, load diameter of phi 10cm, be added with in the chromatographic column of 3kg blank 200-300 order column chromatography silica gel in advance, first use methylene dichloride: methyl alcohol (7:1V/V) wash-out, progressively increase the ratio of methyl alcohol in elutriant again, use methylene dichloride successively: methyl alcohol (7:2V/V), methylene dichloride: methyl alcohol (7:3V/V), methylene dichloride: methyl alcohol (7:4V/V), methylene dichloride: methyl alcohol (7:5V/V), methylene dichloride: the elution of methyl alcohol (7:6V/V), the elutriant of often kind of ratio uses after a certain non-master colour band in chromatographic separation column bottom elutes, change a kind of new ratio elutriant again, until finally use methylene dichloride: the elution of methyl alcohol (1:1V/V), TLC thin layer chromatography detects wash-out process, (silica-gel plate GF254 plate, developping agent is trichloromethane, vinyl acetic monomer, the mixed solution of first alcohol and water, described trichloromethane, vinyl acetic monomer, the volume ratio of first alcohol and water is 15:40:22:10, with ginsenoside Rg1's reference substance for contrast, upright ascending development, 10% sulfuric acid ethanol spraying colour developing, 105 DEG C are heated 10 minutes, sample and reference substance show identical purple plague purpura point.Flow part reception by 250ml/, be eluted to 57-74 when flowing part, occur Rg1 spot, merge, 50 DEG C, pressure obtains Rg1 crude product with Rotary Evaporators decompression and solvent recovery under being the condition of 0.08Mpa, dissolve, under being placed on 8 DEG C of temperature with vinyl acetic monomer, crystallize out, filters, obtains Rg1 sterling.Product yield is 0.8%, and detecting purity through HPLC is 98%.The actual conditions of HPLC is: C18250 × 4.6mm chromatographic column, and be that acetonitrile and the water of 19.5:80.5 is moving phase with volume ratio, flow velocity 1.0ml/min, determined wavelength is 210nm.
The Structure identification of the ginsenoside Rg1 of gained: white powder, mp191-193 DEG C, [α] D
20+ 26.0
0, ultimate analysis C
42h
720
14.2H
20 theoretical value: C60.28, H9.09: experimental value C60.32, H9.06.FD-MS：839[M+K]
+，823[M+Na]
+，821[M+K-H
20]
+，805[M+Na-H
20]
+，677[M+K-162]
+，661[M+Na-162]
+，839[M+2K]
+.EI-MS：m／e
797,770,752,710,525,483,482,465,464,423,422,405,404,389,331,295,289,271,269,229,211,202,187,169,153,109 etc.
1hNMR δ (ppm): 0.95-1.18 (6CH
3) 1.59 Hes
1.99-2.10 (10 × OCOCH
3), 4.79,4.68 (each 1H, J are 7.5Hz), 5.13
deng.The POP data of compound and the POP data of ginsenoside Rg1 basically identical.
Claims (7)
1. from pseudo-ginseng, prepare a ginsenoside Rg1's method, it is characterized in that, concrete steps are:
The first step: fresh Radix Notoginseng is cleaned, cuts into slices, dries, is ground into Radix Notoginseng powder;
Second step: the Radix Notoginseng powder the first step obtained is placed in round-bottomed flask or refluxing extraction device, with organic solvent heating and refluxing extraction at least one times, united extraction liquid; Organic solvent in described second step is methyl alcohol, and the volume ratio of Radix Notoginseng powder and organic solvent is 1:2-3;
3rd step: the extracting solution rotary evaporation recycling design obtained by second step, obtains Radix Notoginseng extract;
4th step: the Radix Notoginseng extract organic solvent dissolution that the 3rd step is obtained, sample is mixed with silica gel for chromatography, silica gel used is 200-300 order column chromatography silica gel, after organic solvent volatilization is clean, silicagel column in dry method, through methylene dichloride and methanol elution gradient, first use methylene dichloride: methyl alcohol is 7:1(V/V in proportion) wash-out, progressively increase the ratio of methyl alcohol in elutriant again, use methylene dichloride successively: methyl alcohol is 7:2(V/V in proportion), methylene dichloride: methyl alcohol is 7:3(V/V in proportion), methylene dichloride: methyl alcohol is 7:4(V/V in proportion), methylene dichloride: methyl alcohol is 7:5(V/V in proportion), methylene dichloride: methyl alcohol is 7:6(V/V in proportion) elution, the elutriant of often kind of ratio uses after a certain non-master colour band in chromatographic separation column bottom elutes, change a kind of new ratio elutriant again, until finally use methylene dichloride: methyl alcohol is 1:1(V/V in proportion) elution, thin-layer chromatography chromatogram detects wash-out process, developping agent is the mixed solution of trichloromethane, vinyl acetic monomer, first alcohol and water, the volume ratio of described trichloromethane, vinyl acetic monomer, first alcohol and water is 15:40:22:10, collect the elution fraction containing ginsenoside Rg1, after recycling design, obtain the crude product containing Rg1,
5th step: by obtain containing the crude product of Rg1, dissolve with vinyl acetic monomer, when being cooled to 5 DEG C-10 DEG C, crystallize out, filters, obtains Rg1 sterling.
2. from pseudo-ginseng, prepare the method for ginsenoside Rg1 as claimed in claim 1, it is characterized in that, the time of each reflux in described second step is 1-3 hour, and the number of times of heating and refluxing extraction is 4-5 time.
3. from pseudo-ginseng, prepare as claimed in claim 1 the method for ginsenoside Rg1, it is characterized in that, the temperature of the rotary evaporation in described 3rd step is 0-50 DEG C, pressure is 0.07-0.1Mpa.
4. from pseudo-ginseng, prepare the method for ginsenoside Rg1 as claimed in claim 1, it is characterized in that, organic solvent in described 4th step is methyl alcohol, and the weightmeasurement ratio of Radix Notoginseng extract and methyl alcohol is 20g:200ml, and Radix Notoginseng extract is 20g:1200g with the weight ratio of mixing sample silica gel.
5. from pseudo-ginseng, prepare the method for ginsenoside Rg1 as claimed in claim 1, it is characterized in that, high effective liquid chromatography for measuring content is used in described 5th step, actual conditions is: C18 250 × 4.6mm chromatographic column, take volume ratio as acetonitrile and the water of 19.5:80.5 be moving phase, flow velocity 1.0ml/min, determined wavelength is 210nm.
6. from pseudo-ginseng, prepare the method for ginsenoside Rg1 as claimed in claim 1, it is characterized in that, in described 4th step, silica gel used is 200-300 order column chromatography silica gel.
7. from pseudo-ginseng, prepare the method for ginsenoside Rg1 as claimed in claim 1, it is characterized in that, the testing conditions of the thin-layer chromatography chromatogram in described 4th step is: adopt GF254 plate, with ginsenoside Rg1's reference substance for contrast, upright ascending development, develops the color with 10% ethanol solution of sulfuric acid spraying or develops the color with iodine vapor, 105 DEG C of heating 10 minutes, showing violet spot during 10% ethanol solution of sulfuric acid, during iodine vapor colour developing, is yellow spotting.
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| CN104693263B (en) * | 2015-04-01 | 2016-08-24 | 江苏省中医药研究院 | A kind of arasaponin compound with anti-tumor activity and preparation method and application |
| CN107595908A (en) * | 2017-09-26 | 2018-01-19 | 云南金七制药有限公司 | A kind of extracting method that notoginsenoside is extracted from fresh pseudo-ginseng |
| CN111072747A (en) * | 2019-12-26 | 2020-04-28 | 王传涛 | Ginsenoside and ultrasonic extraction method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102108091A (en) * | 2009-12-28 | 2011-06-29 | 云南本草精素生物科技有限公司 | Method for preparing two ginsenoside monomers |
| CN102453072A (en) * | 2010-10-26 | 2012-05-16 | 中国医学科学院药物研究所 | Preparation method of ginsenoside Rg1 |
| CN103232515A (en) * | 2013-05-15 | 2013-08-07 | 南京泽朗医药科技有限公司 | Cyclocarya paliurus glucoside I preparation method |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102108091A (en) * | 2009-12-28 | 2011-06-29 | 云南本草精素生物科技有限公司 | Method for preparing two ginsenoside monomers |
| CN102453072A (en) * | 2010-10-26 | 2012-05-16 | 中国医学科学院药物研究所 | Preparation method of ginsenoside Rg1 |
| CN103232515A (en) * | 2013-05-15 | 2013-08-07 | 南京泽朗医药科技有限公司 | Cyclocarya paliurus glucoside I preparation method |
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