CN102293791A - Production method for extracting effective components of gingko leaf - Google Patents
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a production method for extracting effective components of a gingko leaf, which comprises the following steps of: carrying out enzymolysis pretreatment, adding ethanol for reflowing and extracting once; adding glycoside transferase and corresponding saccharide ligand substances and then adding ethanol for reflowing and extracting once; recovering ethanol from the filtered extraction solution and degreasing; carrying out column chromatography and eluting with water and ethanol; and recovering ethanol from an eluent, condensing the eluent in an ointment shape and drying to obtain an extract. The gingko leaf is extracted by using the production method disclosed by the invention, the content of the effective components can be improved by 43-65 percent; and in addition, the production method has the advantages of simpleness in operation, environment friendliness, no toxicity and low production cost.
Description
Technical field
The present invention relates to a kind of production method of extracting effective components in plants, particularly a kind of production method of extracting the effective component of ginkgo leaf that can treat cardiovascular and cerebrovascular disease belongs to the Chinese medicine preparation field.
Background technology
Semen Ginkgo is the most ancient in the world Relict Plant, have the title of " living fossil " and " botanic panda ", and China is the native place of Semen Ginkgo, and ginkgo biloba content accounts for more than 70% of world's total amount.Fruit of ginkgo and leaf all have very high medical value and edibility.According to the Compendium of Material Medica record, " urine reducing ends nebulousurine to Semen Ginkgo for prepared food warming the lung and benefiting QI, relieving asthma and cough; Eat raw and fall expectorant, sterilization, parasite killing ".Folium Ginkgo extract is widely used in and improves cardio and vascular function at present.Semen Ginkgo is to be familiar with by the foreign study person the earliest and one of the Chinese herbal medicine extensively admitted, and its preparation is best-selling in the world plant amedica.
What content was the highest in the Folium Ginkgo is Flavonoid substances, and terpene lactones is the chemical compound of Semen Ginkgo its specific structure complexity, and effective ingredient such as flavonoid that extracts from Folium Ginkgo and terpene lactones have positive effect to treatment coronary heart disease, cerebral thrombosis cardiovascular and cerebrovascular disease.At present, the extracting method of ginkgetin compound has methods such as water extraction, organic solvent extraction, supercritical extraction.The water extraction cost is low, but extracts rate variance, and raw material employing rate is not high; Organic solvent extraction cost height, the impurity content height; Though supercritical fluid method extraction ratio height, but because of apparatus expensive, popularize restricted, the production technology that the CN101708195A patent discloses a kind of Semen Ginkgo extrac on May 19th, 2010, but this technology still exists the effective component of ginkgo leaf extraction ratio not high, defectives such as not environmental protection, and this technology is extracted, and the content of effective component of ginkgo leaf Flavonoid substances is the highest can only to reach 24.5%, therefore, be badly in need of a kind of production method that can effectively improve Flavonoid substances content in the Folium Ginkgo.
Summary of the invention
Shortcomings such as the extraction effective component of ginkgo leaf extraction ratio that prior art exists is low in order to overcome, not environmental protection the invention provides and a kind ofly can effectively improve the production method that effective component of ginkgo leaf extracts ratio.
For achieving the above object, the invention provides following technical scheme: a kind of production method of extracting effective component of ginkgo leaf is characterized in that may further comprise the steps:
(1) enzymolysis Folium Ginkgo: after the Folium Ginkgo raw material powder was broken into coarse powder, with the enzymolysis solution mix homogeneously, enzymolysis was 1～2 day under 30～60 ℃ temperature;
(2) extract effective component of ginkgo leaf: the ginkgo powder that enzymolysis is good adds ethanol, reflux, extract, is 1 time under 70～85 ℃ temperature, add transglucosidase and corresponding glycosyl ligand substance again, add ethanol then, reflux, extract, is 1 time under 50-60 ℃ temperature, filter, reclaim ethanol, defat after merging twice extracting solution; Reclaim ethanol, defat after merging twice extracting solution;
(3) go up column chromatography: with the extracting solution after the defat by macroporous adsorbent resin column chromatography, then successively the water eluting once, with ethanol elution at least three times, after the positive section of hydrochloric acid magnesium powder test eluent one-tenth, merge eluent, reclaim ethanol, be condensed into the paste after drying.
After the Folium Ginkgo raw material powder is broken into granule, increases material and enzymolysis solution and extracted the solvent contacted specific surface area, raw material fully is dissolved in the solvent, can improve extraction ratio.
Temperature has double influence to enzyme, thereby suitable temperature can make enzyme bring into play its maximum activity, also can keep simultaneously its stability preferably, the pair cell wall carries out cracking, causes the dissolution with solvents flavone compound when cell surface spreads, resistance reduces, help to the solvent main diffusion, improve the extraction ratio of Ginkgo total flavones, under 30～60 ℃ temperature, carry out enzymolysis, help in the stability that keeps enzyme, making enzyme bring into play maximum vigor, make extracting effect reach best; Extraction temperature is directly proportional with lixiviate speed, and temperature raises and can reduce the viscosity of alcohol solvent, thereby improves the extraction efficiency of ginkgetin; But the too high heat stability that might destroy flavone of temperature, the probability of impurity stripping also increases, and can reduce extracting effect on the contrary, therefore keeps suitable temperature just to become very important, and it is best extraction temperature that the present invention is controlled at 70～85 ℃ with temperature.
Further, the enzymolysis solution in the described step (1) is the mixture of water and enzyme, and the mass ratio of water and enzyme is 100: 1; The water of enzymolysis solution and the mass ratio of ginkgo leaf powder are 2: 1.
Preferably, described enzyme is the mixture of a kind of and transglucosidase in transglucosidase or cellulase, hemicellulase, pectase, the compound enzyme.
It is in the main cell wall that constitutes that flavone compound in the Folium Ginkgo and terpene lactones mainly are wrapped in the cellulose, independent ethanol extraction method can't make cell wall break fully, cause the stripping of effective component of ginkgo leaf to have suitable resistance, cause extraction effect to be subjected to very big influence.Folium Ginkgo is carried out the enzyme pretreatment, and enzyme can destroy the cell wall of Folium Ginkgo, make cell wall loose, break, thereby can reduce the stripping resistance of effective component of ginkgo leaf chemical compound, the effective ingredient of Folium Ginkgo is fully dissolved, shorten extraction time, strengthen the extraction effect of effective ingredient.Transglucosidase can make some ester dissolubility effective ingredient in the Folium Ginkgo be converted into the hydrophilic composition of band glycosyl and improve recovery rate.Under 30～60 ℃ temperature, Folium Ginkgo is carried out enzymolysis processing, can make the mixture of cellulase or hemicellulase or pectase or compound enzyme and transglucosidase keep good activity, reach the Expected Results of enzymolysis processing.Ginkgo leaf powder behind the enzymolysis is behind the process ethanol extraction, and with respect to the extracting solution that other method obtains, macromolecular substances is less relatively, be convenient to filter, make impurity minimizing in the resin hole, in the chromatography process, can not stop up the resin hole, make chromatography more abundant, impurity separates more thorough.
Further, described reflux, extract, technology, for the first time with 30～70% ethanol extraction 2 hours, the second time was with 30% ethanol extraction 1.5 hours; Ethanol and ginkgo leaf powder mass ratio are 5: 1 for the first time, are 3: 1 for the second time; The transglucosidase that adds and the mass ratio of corresponding glycosyl ligand substance and ginkgo leaf powder are 1～2: 100; Described corresponding glycosyl ligand substance is maltose or glucose.
Extract under 70 ℃-85 ℃ temperature with 30～70% ethanol for the first time, purpose is the abundant extraction that helps effective ingredient in the Folium Ginkgo, and be convenient to enzyme-added processing for the second time, use transglucosidase and glycosyl ligand substance that Folium Ginkgo is handled for the second time in the reflux, extract,, can improve the polarity of flavone aglycone, re-use 30% ethanol on this basis and carry out the extraction second time, further improve the yield of flavonoid glycoside.
Preferably, reflux, extract, adopts multi-function extractor that feed liquid is extracted in the described step (2), can further increase extraction ratio.
Further, described with ethanol respectively eluting be meant for three times for the first time with 30% ethanol, for the second time with 60% ethanol, distinguish eluting with 80% ethanol for the third time.
After extracting solution is crossed post, be eluted to the eluent clarification with purified water, to remove the macromolecular complex in the macroporous resin chromatographic column, removal of contamination, carry out three subgradient eluting with ethanol respectively again, the effective component of ginkgo leaf that is adsorbed on the resin column is fully separated with resin column, strengthened the purity of extracting.
Preferably, the macroporous adsorbent resin column chromatography in the described step (3) adopts ADX-16 type macroporous adsorptive resins, and this resin is U.S.'s ROHM AND HAAS production, and is strong to ginkgetin and lactone selectivity, can effectively reduce the content of ginkgoic acid.
Beneficial effect of the present invention is as follows:
1, with Folium Ginkgo enzymolysis pretreatment before lixiviate, make the abundant stripping of effective ingredient in the Folium Ginkgo, improved lixiviate effectiveness greatly;
2, the present invention adopts extracted many times, and the method for the ethanol three subgradient eluting of reuse different purity behind the pure water eluting helps complete stripping and abundant effective ingredient of resolving Folium Ginkgo, and the active constituent content of extraction is improved greatly;
3, enzymolysis pretreatment of the present invention, extracted many times, enzymolysis, chromatography, stepwise elution can both improve the extraction ratio of effective constituents of Folium Ginkgo respectively, the technical scheme of comprehensive each step, effectively the technology in each stage of control can make the content of active ingredient in the Folium Ginkgo extract improve 43%～65% at last;
4, simple to operate, the environment-protecting asepsis of production method of the present invention, production cost is low.
The specific embodiment
A kind of production method of extracting effective component of ginkgo leaf of the present invention may further comprise the steps:
(1) at first Folium Ginkgo raw material 1000 grams are ground into coarse powder, then 20 gram enzymes are mixed with 2000 gram water, stirring is made into enzymolysis solution, adds the ginkgo powder of pulverizing again, and enzymolysis is 1～2 day under 30～60 ℃ temperature;
(2) ethanol (concentration of alcohol that adds in the production process can be selected between 30～70%) of ginkgo leaf powder adding 5000 grams 30% that above-mentioned enzymolysis is good, under 70～85 ℃ temperature, use 2 hours after-filtration of multi-function extractor reflux, extract,, the maltose that adds 12ml Suhong475 transglucosidase and 80g/L again, with 3000 gram 30% ethanol, 1.5 hours after-filtration of reflux, extract, under 50-60 ℃ temperature, reclaim ethanol, defat after collecting twice extracting solution;
In the production process, the glycosyl ligand substance can also be used glucose; The mass ratio of transglucosidase and corresponding glycosyl ligand substance and ginkgo leaf powder is 1～2: 100, in the production according to the raw-material quality of Folium Ginkgo, corresponding transglucosidase and the glycosyl ligand substance of adding is 1～2: the quality of rational allocation transglucosidase and glycosyl ligand substance in 100 the scope;
(3) go up column chromatography: the extracting solution after the defat is passed through ADX-16 type macroporous adsorbent resin column chromatography, the water eluting is once successively then, reuse 30% ethanol, 60% ethanol, 80% ethanol is eluting respectively, after the positive section of hydrochloric acid magnesium powder test eluent one-tenth, merge three times eluent, reclaim ethanol, be condensed into the paste after drying.
Below be the concrete technological parameter synopsis of each embodiment:
Claims (7)
1. production method of extracting effective component of ginkgo leaf is characterized in that may further comprise the steps:
(1) enzymolysis pretreatment: after the Folium Ginkgo raw material powder was broken into coarse powder, with the enzymolysis solution mix homogeneously, enzymolysis was 1～2 day under 30～60 ℃ temperature;
(2) extract effective component of ginkgo leaf: the ginkgo powder that enzymolysis is good adds ethanol, reflux, extract, is 1 time under 70～85 ℃ temperature, add transglucosidase and corresponding glycosyl ligand substance again, add ethanol then, reflux, extract, is 1 time under 50～60 ℃ temperature, filter, reclaim ethanol, defat after merging twice extracting solution;
(3) go up column chromatography: with the extracting solution after the defat by macroporous adsorbent resin column chromatography, then successively the water eluting once, with ethanol elution at least three times, after the positive section of hydrochloric acid magnesium powder test eluent one-tenth, merge eluent, reclaim ethanol, be condensed into the paste after drying.
2. a kind of production method of extracting effective component of ginkgo leaf according to claim 1 is characterized in that the enzymolysis solution in the described step (1) is the mixture of water and enzyme, and the mass ratio of water and enzyme is 100: 1; The water of described enzymolysis solution and the mass ratio of ginkgo leaf powder are 2: 1.
3. a kind of production method of extracting effective component of ginkgo leaf according to claim 2 is characterized in that, described enzyme is the mixture of a kind of and transglucosidase in transglucosidase or cellulase, hemicellulase, pectase, the compound enzyme.
4. a kind of production method of extracting effective component of ginkgo leaf according to claim 1 is characterized in that, the reflux, extract, technology in the described step (2), and for the first time with 30～70% ethanol extraction 2 hours, the second time was with 30% ethanol extraction 1.5 hours; Described ethanol and ginkgo leaf powder mass ratio are 5: 1 for the first time, are 3: 1 for the second time; The mass ratio of the transglucosidase of described adding and corresponding glucide and ginkgo leaf powder is 1～2: 100; Described corresponding glycosyl ligand substance is maltose or glucose.
5. a kind of production method of extracting effective component of ginkgo leaf according to claim 1 is characterized in that, reflux, extract, adopts multi-function extractor that feed liquid is extracted in the described step (2).
6. a kind of production method of extracting effective component of ginkgo leaf according to claim 1, it is characterized in that, in the described step (3) with ethanol respectively eluting be meant for three times for the first time with 30% ethanol, for the second time with 60% ethanol, distinguish eluting with 80% ethanol for the third time.
7. a kind of production method of extracting effective component of ginkgo leaf according to claim 1 is characterized in that, the macroporous adsorbent resin column chromatography in the described step (3) adopts ADX-16 type macroporous adsorptive resins.
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| CN103638071A (en) * | 2013-12-23 | 2014-03-19 | 湖南科技学院 | Efficient production method of gingko medicinal extract |
| CN107661361A (en) * | 2017-11-17 | 2018-02-06 | 浙江工业大学上虞研究院有限公司 | A kind of process for super-critical extracting of ginko leaves flavone class compound |
| CN107669713A (en) * | 2017-11-17 | 2018-02-09 | 浙江工业大学上虞研究院有限公司 | A kind of extraction process of ginko leaves flavone class compound |
| CN108379371A (en) * | 2018-02-06 | 2018-08-10 | 芜湖市诺康生物科技有限公司 | A kind of ginkgo leaf Radix Salviae Miltiorrhizae Fish oil soft capsule and preparation method thereof |
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| CN103638071A (en) * | 2013-12-23 | 2014-03-19 | 湖南科技学院 | Efficient production method of gingko medicinal extract |
| CN107661361A (en) * | 2017-11-17 | 2018-02-06 | 浙江工业大学上虞研究院有限公司 | A kind of process for super-critical extracting of ginko leaves flavone class compound |
| CN107669713A (en) * | 2017-11-17 | 2018-02-09 | 浙江工业大学上虞研究院有限公司 | A kind of extraction process of ginko leaves flavone class compound |
| CN108379371A (en) * | 2018-02-06 | 2018-08-10 | 芜湖市诺康生物科技有限公司 | A kind of ginkgo leaf Radix Salviae Miltiorrhizae Fish oil soft capsule and preparation method thereof |
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| CN102293791B (en) | 2013-06-12 |
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