CN106727745B - Method for preparing low-acid ginkgo leaf extract - Google Patents
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Abstract
The invention relates to a method for preparing a low-acid ginkgo leaf extract, which comprises the following steps: (1) mixing the crude extract of folium Ginkgo with polyamide resin; (2) supercritical CO2Fluid extraction; (3) adding intoSupercritical CO treatment with modifier2Extracting lactone; (4) gradient eluting lactone and flavone with polyamide resin adsorbed with folium Ginkgo crude extract; (5) concentrating the lactone extract and the eluent; (6) extracting lactone concentrated solution by ethyl acetate; (7) concentrating and drying the extract; (8) precipitating with ethanol to separate out lactone; (9) concentrating the flavone eluate; (10) vacuum drying; (11) mixing lactone and flavone. The preparation method has reasonable step design and simple process, and can effectively extract lactone and flavone while removing ginkgolic acid; overcomes the defects of the prior art that the process is complicated, and the ginkgolic acid can not be efficiently removed while the lactone and the flavone are extracted.
Description
Technical Field
The invention relates to a method for extracting and separating effective components of traditional Chinese medicines, in particular to a method for preparing a low-acid ginkgo leaf extract.
Background
Ginkgo biloba, also known as ginkgo leaf, is the leaf of ginkgo biloba of the genus Ginkgo of the family Ginkgoaceae. Gingko is a specific species in China, namely gymnospermum, deciduous tree, is one of the oldest middle-aged writings rare plant in China, is called as 'activating stone', and accounts for about 70% of the total amount of the world. Although ginkgo biloba has a long history as a medicine, the study on chemical components and pharmacological actions of ginkgo biloba by scholars at home and abroad has been comprehensively and systematically found since the 60 s of the 20 th century: ginkgo has more chemical components, and at present, more than 100 chemical components are found, wherein the chemical components mainly comprise flavonoid compounds, terpene lactones and the like.
Research shows that the ginkgo leaves contain more than 30 flavonoid compounds, are excellent oxygen radical scavengers and lipid antioxidants and have outstanding efficacy on treating and preventing cardiovascular and cerebrovascular diseases.
The ginkgolide terpene compounds in folium Ginkgo are not found in other plants so far, have unique physiological action and therapeutic action, are antagonists of platelet activating factor, and especially have good effects in preventing and treating asthma, lung anaphylaxis, anticoagulation and preventing cerebral thrombosis.
Ginkgolic acid is an organic acid contained in ginkgo leaves, is an alkyl acid or alkylphenol compound with 13-17 carbon atoms, is considered to have potential sensitization and mutation effects and strong cytotoxicity, and can cause severe anaphylactic reaction, gene mutation and nerve injury to cause adverse reactions such as nausea, heartburn, anaphylactic shock, allergic purpura, exfoliative dermatitis and the like. From the viewpoint of drug safety, the german Schwabe company proposed in the patent in 1991 that the content of ginkgolic acid compounds is required to be less than 10ppm, the german ministry of health proposed in 1997 that the content of ginkgolic acid should be less than 5ppm, and if the preparation can not reach the standard, the registration number and the right to sell thereof are forcibly cancelled, and the ginkgo biloba extract standard published in the chinese pharmacopoeia 2010 requires that the content of flavonol glycosides is more than 24%, the content of terpene lactones is more than 6%, the content of ginkgolic acid is less than 5ppm, and the water solubility is more than 5%. Therefore, the reduction of the ginkgolic acid amount while extracting the flavone and the lactone becomes a technical problem which is overcome firstly.
The production process of the low-acid ginkgo leaf extract has various processes, and the chemical structure and the material property of the ginkgoic acid can be studied. For example, ginkgolic acids can be removed by a method of forming a precipitate by the carboxyl group of ginkgolic acids and soluble lead salt, but the lead salt is extremely toxic, has strict requirements on product quality indexes and environmental discharge, and is difficult to form a precipitate with alkylphenol containing no carboxyl group, so the method is not applicable; the polarity of the ginkgoic acid is greatly weakened due to the existence of the ginkgoic acid long-chain alkyl group, the water solubility is poor, so the ginkgoic acid can be separated out by a method of cooling and settling an extracting solution, the invention patent CN102335209A discloses a preparation method and a product of an ultra-low acid water-soluble ginkgo leaf extract, the content of the ginkgoic acid is reduced by using a method of combining flocculation precipitation with refrigeration, but the method is rough, the selectivity is not too high, and the active ingredients are lost while the ginkgoic acid is reduced; the invention patent CN106038619A discloses a ginkgo leaf extract and a preparation method thereof, which adopts supercritical CO2The extraction and purification are organically combined with a macroporous resin method, and supercritical CO is firstly adopted2The extraction method is used for initial extraction, and then purification is carried out by a macroporous resin method, wherein although low-temperature cooling is mentioned to remove part of the ginkgo biloba acid, the strict control is not specially carried out on the content of the ginkgo biloba acid.
Disclosure of Invention
The invention aims to solve the technical problem of the prior art and provides a method for preparing a low-acid ginkgo leaf extract, which is simple in process, can effectively remove ginkgoic acid while efficiently extracting flavone and lactone.
The technical scheme adopted by the invention for solving the technical problems is as follows: a method for preparing a low-acid ginkgo leaf extract is characterized by comprising the following steps:
(1) mixing the ginkgo leaf crude extract with a carrier: dissolving the ginkgo leaf crude extract by using 50-55 ℃ absolute ethyl alcohol, filtering, mixing the filtrate with polyamide resin, wherein the mass ratio of the added polyamide resin to the ginkgo leaf crude extract is 1-3: 1;
(2) supercritical CO2And (3) extraction: putting the mixture obtained in the step (1) into an extraction kettle, sealing the extraction kettle, setting the extraction temperature to be 50-65 ℃, the pressure to be 25-40 MPa, and CO2The flow rate is 2-3L/min, the extraction time is 1-3 hours, and CO is introduced2Extracting, and collecting a product in the separation kettle;
(3) adding a modifier for extraction: introducing an ethanol modifier with the volume fraction of 20-28% for supercritical CO2Extracting for 1-3 hours, and collecting a product in the separation kettle;
(4) resin gradient elution: subjecting the mixture obtained in the step (3) to supercritical CO2Performing dry column packing on the extracted polyamide resin adsorbed with the ginkgo leaf crude extract, performing gradient elution by using an ethanol water solution, eluting by using an ethanol water solution with the volume fraction of 15-20%, eluting by using an ethanol water solution with the volume fraction of 72-78%, and collecting the eluent;
(5) concentration: concentrating the low-concentration eluted eluent obtained in the step (4) and the product in the separation kettle in the step (3) under reduced pressure, wherein the temperature of the concentration under reduced pressure is 50-60 ℃, and the pressure is 0.07-0.09 MPa;
(6) and (3) extraction: extracting the concentrated solution obtained in the step (5) with ethyl acetate for 3-4 times;
(7) concentrating and drying: concentrating the extract obtained in the step (6) under reduced pressure, evaporating the concentrate to dryness in a water bath, wherein the temperature of the concentration under reduced pressure is 50-60 ℃, and the pressure is 0.07-0.09 MPa;
(8) alcohol precipitation: adding a small amount of absolute ethyl alcohol into the dried product obtained in the step (7) for dissolving, and standing for 36-60 hours at the temperature of-15 to-25 ℃ to separate out flocculent precipitates;
(9) concentration: concentrating the high-concentration eluted eluent obtained in the step (4) under reduced pressure, wherein the temperature of the concentration under reduced pressure is 50-60 ℃, and the pressure is 0.07-0.09 MPa;
(10) and (3) vacuum drying: respectively carrying out vacuum drying on the precipitate obtained in the step (8) and the concentrated solution obtained in the step (9) at the temperature of 70-80 ℃ and under the pressure of 0.08-0.09 MPa to obtain target products of lactone and flavone;
(11) mixing: and (4) mixing the lactone obtained in the step (10) with flavone to obtain a target product.
The ginkgolic acid is 6-alkyl salicylic acid, which has polar groups such as aromatic hydroxyl and carboxyl in the structure, but the side chain at the 6 position is a long chain with 13-17 carbon atoms, so that the ginkgolic acid is fat-soluble, shows weak polarity, and can be dissolved in nonpolar supercritical CO2In (1). The main medicinal components of flavone and lactone in folium Ginkgo have high polarity, and supercritical CO can not be used without adding polar agent2And (4) extracting. So that supercritical CO is utilized2Not only can selectively extract ginkgolic acid in the ginkgo leaves, but also has no influence on effective components of flavone and lactone in the ginkgo leaves.
Supercritical CO with modifier2In the fluid, the solubility of flavone is higher than that of lactone under the same pressure, so that lactone is preferentially separated out after pressure reduction, while most of flavone is separated out during resin gradient elution, and supercritical CO is adopted2The fluid extraction is combined with the resin gradient elution to realize the refining and purification of the lactone and the flavone.
Preferably, in the step (1), the crude ginkgo biloba extract is dissolved by absolute ethyl alcohol at 52 ℃, and the mass ratio of the added polyamide resin to the crude ginkgo biloba extract is 1: 1.
preferably, the temperature of the extraction in the step (2) is 55 ℃, the pressure is 30MPa, and CO is2The flow rate was 2.5L/min and the extraction time was 1 hour.
Preferably, the modifier introduced in the step (3) is ethanol water with the volume fraction of 25%, the extraction time is 2.5 hours, the flavone is not substantially extracted under the condition, and the lactone is substantially enriched in the separation kettle.
Preferably, the gradient elution in the step (4) is that the elution is firstly carried out by using 17% ethanol water solution by volume fraction, and then is carried out by using 75% ethanol water solution by volume fraction. According to the principle of similarity and compatibility, lactone is eluted by low-concentration ethanol water solution, and flavone is eluted by high-concentration ethanol water solution.
Preferably, the temperature for concentration under reduced pressure in the step (5) and the step (9) is 55 ℃ and the pressure is 0.08 MPa.
Preferably, the concentrated solution in the step (6) is extracted three times with ethyl acetate.
Preferably, the temperature for concentration under reduced pressure in the step (7) is 55 ℃ and the pressure is 0.08 MPa.
Preferably, the dried product in the step (8) is dissolved in absolute ethanol and then left to stand at-18 ℃ for 48 hours.
Preferably, the temperature of the vacuum drying in the step (10) is 80 ℃ and the pressure is 0.09 MPa.
Compared with the prior art, the invention has the advantages that:
1. the invention adopts supercritical CO2The extraction and polyamide resin column-passing combination method adopts supercritical CO firstly under the condition of no adding modifier2Removing ginkgolic acid by extraction, adding modifier to primarily separate lactone and flavone, and supercritical CO extraction2Gradient elution is carried out on the extracted polyamide resin absorbed with the ginkgo leaf crude extract, and lactone and flavone are further separated and purified, the method effectively extracts the lactone and the flavone and simultaneously removes ginkgoic acid, and the ginkgoic acid content of the prepared ginkgo leaf extract is lower than 5 ppm;
the method for preparing the low-acid ginkgo leaf extract has reasonable step design and simple process, and overcomes the defects of complicated process and incapability of efficiently removing ginkgolic acid while extracting lactone and flavone in the prior art;
the low acid in the invention means that the ginkgolic acid content is lower than 5ppm, and the requirement of the standard of the ginkgo leaf extract in Chinese pharmacopoeia is met.
2. Selecting polyamide resin as carrier, mixing folium Ginkgo crude extract and polyamide resin, extracting in extraction tank, and subjecting to supercritical CO extraction2The extracted flavone is easier to process into a high-concentration product, and simultaneously, the flavone and the lactone are further separated.
3. The supercritical extraction steps are less, the sample treatment time is greatly shortened, the solvent is saved, and organic solvents except ethanol are not used in the process.
4. The supercritical extraction is carried out under high pressure, and Escherichia coli, etc. hardly survive, thereby improving the quality of folium Ginkgo extract and increasing the competitive power of the product.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example 1
Dissolving 100g of folium Ginkgo crude extract with 50 deg.C anhydrous ethanol, filtering, mixing the filtrate with polyamide resin (the weight ratio of folium Ginkgo crude extract to polyamide resin is 1:1), and adding into extraction kettle. Sealing the extraction kettle, setting the extraction temperature at 50 ℃, the pressure at 25MPa and CO2The flow rate is 2L/min; opening of CO2Steel cylinder, CO2Condensing the gas in a condenser, pumping the condensed gas into an extraction kettle, extracting for 3 hours, and performing supercritical CO extraction2The fluid carrying the extract flows out from the top of the kettle, and enters a separation kettle after decompression, the extract is separated out, and the product (ginkgolic acid) in the separation kettle is collected. Then introducing 20% ethanol modifier for supercritical CO2Extracting for 1 hour, collecting the product (more lactone and less flavone) in the separation kettle; then subjecting the above to supercritical CO2Loading the extracted polyamide resin adsorbed with folium Ginkgo crude extract into column by dry method, eluting lactone with 15% ethanol water solution until eluate is colorless, eluting flavone with 72% ethanol water solution, and collecting by stages. Mixing the 15% ethanol eluate and 20% ethanol water solution as modifier, and concentrating under reduced pressure to about 150ml at 50 deg.C and 0.07 MPa; 600ml of concentrated solutionExtracting with ethyl acetate for four times (200ml, 100ml), concentrating the extractive solution at 50 deg.C under 0.07MPa, evaporating to dryness in water bath, adding small amount of anhydrous ethanol for dissolving, standing at-15 deg.C for 60 hr to separate out a large amount of flocculent precipitate, vacuum filtering, washing off yellow precipitate with small amount of anhydrous ethanol, and oven drying at 70 deg.C under 0.08MPa to obtain loose white lactone with purity of 51%. Concentrating 72% ethanol eluate under reduced pressure at 50 deg.C and 0.07MPa, and oven drying at 70 deg.C and 0.08MPa to obtain flavone with purity of 30.3%. Mixing the obtained lactone and flavone to obtain a qualified product: 24.9 percent of flavone, 6.1 percent of lactone and less than 5ppm of ginkgolic acid.
Example 2
Dissolving 100g of folium Ginkgo crude extract with 52 deg.C anhydrous ethanol, filtering, mixing the filtrate with polyamide resin (the mass ratio of folium Ginkgo crude extract to polyamide resin is 1:2), and adding into extraction kettle. Sealing the extraction kettle, setting the extraction temperature at 55 ℃, the pressure at 30MPa and CO2The flow rate is 2.5L/min; opening of CO2Steel cylinder, CO2Condensing the gas in a condenser, pumping the condensed gas into an extraction kettle, extracting for 2 hours, and performing supercritical CO extraction2The fluid carrying the extract flows out from the top of the kettle, and enters a separation kettle after decompression, the extract is separated out, and the product (ginkgolic acid) in the separation kettle is collected. Then introducing 25% ethanol modifier for supercritical CO2Extracting for 2.5 hours, collecting the product (more lactone and less flavone) in the separation kettle; then subjecting the above to supercritical CO2Loading the extracted polyamide resin adsorbed with folium Ginkgo crude extract into column by dry method, eluting lactone with 17% ethanol water solution until eluate is colorless, eluting flavone with 75% ethanol water solution, and collecting by stages. Mixing the 17% ethanol eluate and 25% ethanol water solution as modifier, and concentrating under reduced pressure to about 175ml at 55 deg.C and 0.08 MPa; extracting the concentrated solution with 500ml ethyl acetate for three times (200ml, 100ml), concentrating the extractive solution at 55 deg.C under 0.08MPa, evaporating in water bath, dissolving with small amount of anhydrous ethanol, and standing at-18 deg.CPrecipitating a large amount of flocculent precipitate in 48 hr, vacuum filtering, washing off yellow precipitate with small amount of anhydrous ethanol, and oven drying at 75 deg.C under 0.09Mpa to obtain loose white lactone with purity of 60%. Concentrating 75% ethanol eluate under reduced pressure at 55 deg.C and 0.08MPa, and oven drying at 75 deg.C and 0.09MPa to obtain flavone with purity of 38.2%. Mixing the obtained lactone and flavone to obtain a qualified product: 35.9 percent of flavone, 7.3 percent of lactone and less than 5ppm of ginkgolic acid.
Example 3
Dissolving 100g of folium Ginkgo crude extract with 55 deg.C anhydrous ethanol, filtering, mixing the filtrate with polyamide resin (the weight ratio of folium Ginkgo crude extract to polyamide resin is 1:3), and adding into extraction kettle. Sealing the extraction kettle, setting the extraction temperature at 65 ℃, the pressure at 40MPa and CO2The flow rate is 3L/min; opening of CO2Steel cylinder, CO2Condensing the gas in a condenser, pumping the condensed gas into an extraction kettle, extracting for 1 hour, and performing supercritical CO extraction2The fluid carrying the extract flows out from the top of the kettle, and enters a separation kettle after decompression, the extract is separated out, and the product (ginkgolic acid) in the separation kettle is collected. Then introducing 28% ethanol modifier for supercritical CO2Extracting for 3 hours, collecting the product (more lactone and less flavone) in the separation kettle; then subjecting the above to supercritical CO2Loading the extracted polyamide resin adsorbed with folium Ginkgo crude extract into column by dry method, eluting lactone with 20% ethanol water solution until eluate is colorless, eluting flavone with 78% ethanol water solution, and collecting by stages. Mixing the 20% ethanol eluate and 28% ethanol water solution as modifier, and concentrating under reduced pressure to about 200ml at 60 deg.C under 0.09 MPa; extracting the concentrated solution with 500ml ethyl acetate for three times (200ml, 100ml), concentrating the extractive solution at 60 deg.C under 0.09MPa, evaporating to dryness in water bath, adding small amount of anhydrous ethanol for dissolving, standing at-25 deg.C for 36 hr to separate out a large amount of flocculent precipitate, vacuum filtering, washing off yellow precipitate with small amount of anhydrous ethanol, oven drying at 80 deg.C under 0.09MPa to obtain loose white lactone with purity of 54 percent. Concentrating 78% ethanol eluate under reduced pressure at 60 deg.C and 0.09MPa, and oven drying at 80 deg.C and 0.09MPa to obtain flavone with purity of 33.7%. Mixing the obtained lactone and flavone to obtain a qualified product: 26.8 percent of flavone, 6.9 percent of lactone and less than 5ppm of ginkgolic acid.
Example 4
100g of crude ginkgo leaf extract is put into an extraction kettle. Sealing the extraction kettle, setting the extraction temperature at 55 ℃, the pressure at 30MPa and CO2The flow rate is 2L/min; opening of CO2Steel cylinder, CO2Condensing the gas in a condenser, pumping the condensed gas into an extraction kettle, introducing an ethanol modifier, extracting for 1 hour, and performing supercritical CO extraction2The extract is carried out of the top of the kettle, and enters a separation kettle after being decompressed, the extract is separated out, and the product in the separation kettle is collected. Concentrating the collected solution under reduced pressure until no alcohol smell exists, wherein the temperature of the concentration under reduced pressure is 60 ℃, and the pressure is 0.09 MPa; drying the concentrated solution at 80 deg.C under 0.09Mpa to obtain the final product. The ginkgo biloba extracts extracted by the ethanol modifiers with different concentrations are shown in the following table:
as can be seen from the above table, under 30MPa, the separating effect of the modifier with different concentrations on flavone and lactone in the crude extract of ginkgo leaf is different, especially the supercritical CO modified by 25% ethanol2When extraction is carried out, the separation effect is best, the flavone is not extracted basically, and the lactone is greatly enriched in the separation kettle, and the content is as high as 10.43%, which is very beneficial to further purifying the flavone and the lactone.
Claims (9)
1. A method for preparing a low-acid ginkgo leaf extract is characterized by comprising the following steps:
(1) mixing the ginkgo leaf crude extract with a carrier: dissolving the ginkgo leaf crude extract by using 50-55 ℃ absolute ethyl alcohol, filtering, mixing the filtrate with polyamide resin, wherein the mass ratio of the added polyamide resin to the ginkgo leaf crude extract is 1-3: 1;
(2) supercritical CO2And (3) extraction: putting the mixture obtained in the step (1) into an extraction kettle, sealing the extraction kettle, setting the extraction temperature to be 50-65 ℃, the pressure to be 25-40 MPa, and CO2The flow rate is 2-3L/min, the extraction time is 1-3 hours, and CO is introduced2Extracting, and collecting a product in the separation kettle;
(3) adding a modifier for extraction: introducing 25% ethanol modifier for supercritical CO2Extracting for 2.5 hours, and collecting a product in the separation kettle;
(4) resin gradient elution: subjecting the mixture obtained in the step (3) to supercritical CO2Performing dry column packing on the extracted polyamide resin adsorbed with the ginkgo leaf crude extract, performing gradient elution by using an ethanol water solution, eluting by using an ethanol water solution with the volume fraction of 15-20%, eluting by using an ethanol water solution with the volume fraction of 72-78%, and collecting the eluent;
(5) concentration: concentrating the low-concentration eluted eluent obtained in the step (4) and the product in the separation kettle in the step (3) under reduced pressure, wherein the temperature of the concentration under reduced pressure is 50-60 ℃, and the pressure is 0.07-0.09 MPa;
(6) and (3) extraction: extracting the concentrated solution obtained in the step (5) with ethyl acetate for 3-4 times;
(7) concentrating and drying: concentrating the extract obtained in the step (6) under reduced pressure, evaporating the concentrate to dryness in a water bath, wherein the temperature of the concentration under reduced pressure is 50-60 ℃, and the pressure is 0.07-0.09 MPa;
(8) alcohol precipitation: adding a small amount of absolute ethyl alcohol into the dried product obtained in the step (7) for dissolving, and standing for 36-60 hours at the temperature of-15 to-25 ℃ to separate out flocculent precipitates;
(9) concentration: concentrating the high-concentration eluted eluent obtained in the step (4) under reduced pressure, wherein the temperature of the concentration under reduced pressure is 50-60 ℃, and the pressure is 0.07-0.09 MPa;
(10) and (3) vacuum drying: respectively carrying out vacuum drying on the precipitate obtained in the step (8) and the concentrated solution obtained in the step (9) at the temperature of 70-80 ℃ and under the pressure of 0.08-0.09 MPa to obtain target products of lactone and flavone;
(11) mixing: and (4) mixing the lactone obtained in the step (10) with flavone to obtain a target product.
2. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: dissolving the ginkgo leaf crude extract in the step (1) by using absolute ethyl alcohol at 52 ℃, wherein the mass ratio of the added polyamide resin to the ginkgo leaf crude extract is 1: 1.
3. the method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: the temperature of the extraction in the step (2) is 55 ℃, the pressure is 30MPa, and CO is2The flow rate was 2.5L/min and the extraction time was 1 hour.
4. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: and (4) eluting with gradient in the step (4), namely eluting with 17% ethanol water solution by volume fraction, and then eluting with 75% ethanol water solution by volume fraction.
5. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: the temperature for decompression concentration in the step (5) and the step (9) is 55 ℃, and the pressure is 0.08 MPa.
6. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: and (3) extracting the concentrated solution in the step (6) with ethyl acetate for three times.
7. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: the temperature of the reduced pressure concentration in the step (7) is 55 ℃, and the pressure is 0.08 MPa.
8. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: and (3) dissolving the dried product in the step (8) by using absolute ethyl alcohol, and standing for 48 hours at the temperature of-18 ℃.
9. The method for preparing a low acid ginkgo leaf extract according to claim 1, wherein: the temperature of vacuum drying in the step (10) is 80 ℃, and the pressure is 0.09 MPa.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1172669A (en) * | 1997-08-22 | 1998-02-11 | 孙云鹏 | Method for extracting and separating flavone and terlactone from ginkgo leaf by supercritical carbon dioxide |
| CN1172670A (en) * | 1997-08-22 | 1998-02-11 | 孙云鹏 | Method for extracting flavone and terlactone from ginkgo leaf by supercritical carbon dioxide |
| CN102416027A (en) * | 2011-12-01 | 2012-04-18 | 徐州医学院 | Gingkgo active ingredient extraction and separation method and use of extracted active matters in medicines for treating cardiovascular and cerebrovascular diseases |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1172669A (en) * | 1997-08-22 | 1998-02-11 | 孙云鹏 | Method for extracting and separating flavone and terlactone from ginkgo leaf by supercritical carbon dioxide |
| CN1172670A (en) * | 1997-08-22 | 1998-02-11 | 孙云鹏 | Method for extracting flavone and terlactone from ginkgo leaf by supercritical carbon dioxide |
| CN102416027A (en) * | 2011-12-01 | 2012-04-18 | 徐州医学院 | Gingkgo active ingredient extraction and separation method and use of extracted active matters in medicines for treating cardiovascular and cerebrovascular diseases |
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