CN102464697A - Method for combined preparation of high purity ursolic acid and high purity oleanolic acid from Ligustrum lucidum - Google Patents
Method for combined preparation of high purity ursolic acid and high purity oleanolic acid from Ligustrum lucidum Download PDFInfo
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- CN102464697A CN102464697A CN201010539933XA CN201010539933A CN102464697A CN 102464697 A CN102464697 A CN 102464697A CN 201010539933X A CN201010539933X A CN 201010539933XA CN 201010539933 A CN201010539933 A CN 201010539933A CN 102464697 A CN102464697 A CN 102464697A
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Abstract
The invention which discloses a method for the combined preparation of high purity ursolic acid and high purity oleanolic acid from Ligustrum lucidum belongs to the technical field of natural medicines, wherein the method comprises processes for extracting and purifying ursolic acid and oleanolic acid from Ligustrum lucidum leaves or Ligustrum lucidum seeds. The method adopted in the invention allows the cheap and easily available Ligustrum lucidum leaves or Ligustrum lucidum seeds which are treated as a raw material to be extracted by refluxing with ethanol. The treatment for "three wastes" has the advantages of small capacity and easiness, ethanol can be recycled, and waste residue residual after the extraction can be used as a fertilizer, so the method basically generates no pollution to environment, and the cost of the obtained ursolic acid is low. In addition, the method of the invention is simple and fast. The obtained ursolic acid is a raw material of medicines for blood fat reduction and cardiovascular disease treatment; and the obtained oleanolic acid is an auxiliary hepatopathy medicine recorded by a nonprescription medicine directory in China. The method is suitable for the industrialization large-scale production.
Description
Technical field
The invention belongs to natural medicine technical field, be specifically related to a kind of also method of its main active component ursolic acid of purifying, Oleanolic Acid of from glossy privet, extracting.
Technical background
Glossy privet is an Oleaceae ligustrum plant, and formal name used at school is Ligustrum Lucidum Ait, and it is the four seasons evergreen big shrubs or dungarunga, and the whole world is had an appointment 50 kinds; Just there are 38 kinds in China, are distributed widely in East China, south China; Southwest and Central China, plant resources is abundant, and distributed pole is wide.
Along with going deep into of pharmacological research, find that the ligustrum plant has multiple physiologically active, but also also insufficient to the Chemical Composition research of ligustrum plant.For a long time, the position that we are studied to the ligustrum plant utilizes not thorough, is not used as medicine basically, wastes very big.
(Ursolic acid UA), has another name called ursonic acid to ursolic acid, and urson, α-amyrin are a kind of slightly acidic pentacyclic triterpenoids.Very wide in distributed in nature, be the functional component of multiple natural product.The pure article of ursolic acid are white, needle-shaped crystals (crystallization in the ethanol), and bitter, its basic framework are the five rings parent nucleus of polyoxy pinane.Chemical name (3 β)-3-Hydroxyurs-12-en-28-oic acid, molecular formula C30 H48O3, relative molecular mass 456.68; Fusing point 285-291 ℃; Be soluble in dioxane, pyridine, dissolve in methyl alcohol, ethanol, butanols, butanone, slightly be dissolved in acetone; Be slightly soluble in benzene, chloroform, ether, water insoluble and sherwood oil.UA is wider in distributed in nature, exists as in plants such as Sambucus chinensis, Loquat Leaf, black bearberry, Glossy Privet Fruit, hawthorn, psyllium, Spica Prunellae, Spreading Hedyotis Herb, existing or be combined into glucoside with sugar with free form.Ursolic acid has multiple biological activity, and Japan and other countries is applied to it in food as natural antioxidants.
Oleanolic Acid is the isomers of ursolic acid, claims oleanolic acid again, has heat-clearing, anti-inflammatory, cardiac stimulant, hypoglycemic, and reducing blood-fat, inhibition transformation reactions, cholagogic and removing jaundice isoreactivity are for protecting the liver good medicine.
Ursolic acid and Oleanolic Acid extensively are present in the leaf and fruit of specific section genus; Different starting material separation and purification obtains higher ursolic acid of purity or Oleanolic Acid; The main means such as column chromatography or macroporous resin enrichment that adopt will obtain the purity high product to physico-chemical property extremely similar ursolic acid and Oleanolic Acid respectively, and difficulty is still bigger; Although column chromatography can separate ursolic acid and approximate impurity well; But this method applied sample amount is little, and yield is low, is unfavorable for large-scale industrial production.Eluent need be used organic solvents such as sherwood oil, benzene, chloroform, methyl alcohol, ETHYLE ACETATE simultaneously, and cost is big, is unfavorable for production safety, also environment is worked the mischief.All contain a certain amount of ursolic acid and Oleanolic Acid in Leaf of Glossy Privet and the Glossy Privet Fruit.It is main that the process for extracting of having reported mainly combines certain separation means with the organic solution lixiviate; For example CN1358733 obtains the ursolic acid in the Leaf of Glossy Privet with the method that alcohol reflux, activated carbon decolorizing, drying etc. combine; Though it is easier to be somebody's turn to do invention, can only obtain the ursolic acid bullion.
Summary of the invention:
Technical problem to be solved by this invention is to overcome the weak point of aforesaid method, and low, the environmental friendliness of a kind of cost of research and design is convenient to the simple and easy method for preparing high-purity ursolic acid and Oleanolic Acid of suitability for industrialized production.Mainly be to utilize ursolic acid and Oleanolic Acid in water, not to dissolve, different PH solubility property different characteristic design in ethanol.Yield is high, and product is pure.Product is through Spectrum Analysis, the conformance to standard of its result and ursolic acid and Oleanolic Acid.
Technical scheme provided by the invention is: a kind of from Leaf of Glossy Privet or Glossy Privet Fruit the method for extraction and separating and purifying ursolic acid, Oleanolic Acid.Said glossy privet comprises Leaf of Glossy Privet or Glossy Privet Fruit, and extraction and separation purification method comprise the following steps: A) glossy privet blade or Glossy Privet Fruit are boiled with buck, filtered while hot, alkali compound is used hot wash, removes content of wax impurity.With the dregs of a decoction that obtain is extracting solution with the pure water, in the boiling water water-bath, extracts, and filters, and removes water-soluble impurities such as carbohydrate, tannin.With the filter residue that filtration obtains, be solvent with the ethanolic soln, the refluxing extraction after-filtration obtains ethanol extract in water-bath; B) ethanol extract is used activated carbon decolorizing, the filtering gac is regulated about ethanol extract pH value to 11; After leaving standstill impurity screening filtrate decompression is concentrated into small volume (concentrating the 1/3-1/4 of front volume), has a large amount of insolubless to separate out, filter; Leaching thing is the Oleanolic Acid bullion, and filtrating is regulated the pH value to 3-4, adds isopyknic pure water; Place 4 ℃ to spend the night, leach insolubles, obtain the ursolic acid bullion; C) the Oleanolic Acid bullion is washed with pure water, insolubles is used anhydrous alcohol solution, and regulating PH is 11, adds activated carbon decolorizing, places and filters; Filtrating PH transfers to 1, separates out throw out, and throw out is boiled with buck, filters, and repeats 2-3 time; Insolubles is used boiling pure water, and filtered while hot must be washed thing, will wash thing and use anhydrous alcohol solution, activated carbon decolorizing once more; Filter, filtrating PH transfers to 1, separates out throw out, and the zero(ppm) water that boils washing can obtain highly purified Oleanolic Acid.D) the ursolic acid bullion is washed with the 1%NaOH aqueous solution, then washed, use absolute ethyl alcohol recrystallization twice then, use methanol mixing solutions or acetate ethanol mixing solutions recrystallization at last, promptly obtain highly purified ursolic acid with pure water.
The present invention adopts aforesaid method, is that raw material is used alcohol reflux with Nuzhen, an ancient nationality in China, ancestors of the Manchus, who founded the Jin Dynasty's leaf cheap and easy to get or Glossy Privet Fruit." three wastes " treatment capacity is little and easy, and the recyclable usefulness again of ethanol is extracted the last waste residue in back and can be used as fertilizer.Thereby the present invention do not produce pollution basically to environment, and the ursolic acid cost that is obtained is lower.In addition, method of the present invention is easy, quick.The ursolic acid that obtains is reducing blood-fat, the medical material of treatment cardiovascular diseases, and Oleanolic Acid is the hepatopathy adjuvant drug that national nonprescription drugs catalogue is recorded.Be fit to batch production scale operation.
Embodiment:
Embodiment 1:
Get Leaf of Glossy Privet dry product 1Kg, add 5% aqueous sodium hydroxide solution 8L, boiled 30 minutes; The filtering alkaline solution is used the hot wash dregs of a decoction while hot, residue is joined boiled in the 10L water 1.5 hours; The water cooking liquid that inclines, residue divides refluxing extraction 3 times with 95% ethanol, and each amount of ethanol is 6L; Merge alcohol extract, add the gac of alcohol extract volume 0.3%, stir decolouring 1 hour down at 50-60 ℃; The filtering gac, it is about 11 that the decolouring alcohol extract uses the 1mol/L aqueous sodium hydroxide solution to transfer to PH, leaves standstill 12 hours after-filtration, removes deposition, filtrate decompression is concentrated into about 1/3 (5L) that do not concentrate front volume; Have a large amount of insolubless to separate out, this insolubles is the Oleanolic Acid bullion, filters, and filtrating is regulated PH between 3-4 with the 6mol/L aqueous hydrochloric acid.In filtrating, add 5L water, place 4 ℃ and spend the night, have a large amount of depositions to separate out.Filter, precipitate is the ursolic acid bullion.
The purifying of Oleanolic Acid:
The above-mentioned Oleanolic Acid bullion 10.4g that obtains is washed with an amount of pure water; Water-insoluble is used the 175ml anhydrous alcohol solution, regulates PH to 11 with 10% aqueous sodium hydroxide solution, adds the 3g gac; Placed 50-60 ℃ of stirring in water bath 15 minutes; The filtering gac, filtrating is regulated PH to 1 with the 6mol/L aqueous hydrochloric acid, separates out throw out 9.6g.Throw out is joined 60ml, boiled in 5% aqueous sodium hydroxide solution 30 minutes, filter, insolubles is boiled the pure water washing with 60ml; Filtered while hot must be washed thing 8.1g, will wash thing and use the 80ml anhydrous alcohol solution, adds the 0.8g gac; Placed 50-60 ℃ of stirring in water bath 15 minutes, and filtered, filtrating is regulated PH to 1 with the 6mol/L aqueous hydrochloric acid; Separate out Oleanolic Acid, filter, the zero(ppm) water that boils washing obtains Oleanolic Acid 4.7g.Product is through Spectrum Analysis, its result and Oleanolic Acid conformance to standard.
The purifying of ursolic acid:
The above-mentioned ursolic acid bullion 42.8g that obtains is washed with an amount of 1% sodium hydroxide solution; Then with an amount of pure water washing; With insolubles 37.9g with an amount of absolute ethyl alcohol recrystallization twice, use again the 200ml volume ratio be 1: 1 the crystallization of methanol mixing solutions once, finally obtain ursolic acid 19.3g.Product is through Spectrum Analysis, its result and ursolic acid conformance to standard.
Embodiment 2:
Get Glossy Privet Fruit dry product 1Kg, add 5% aqueous sodium hydroxide solution 8L, boiled 30 minutes; The filtering alkaline solution is used the hot wash dregs of a decoction while hot, residue is joined boiled in the 10L water 2 hours; The water cooking liquid that inclines, residue divides refluxing extraction 3 times with 90% ethanol, and each amount of ethanol is 6L; Merge alcohol extract, add the gac of alcohol extract volume 0.3%, stir decolouring 1 hour down at 50-60 ℃; The filtering gac, it is about 11 that the decolouring alcohol extract uses the 1mol/L aqueous sodium hydroxide solution to transfer to PH, leaves standstill 12 hours after-filtration, removes deposition, filtrate decompression is concentrated into about 1/3 (5L) that do not concentrate front volume; Have a large amount of insolubless to separate out, this insolubles is the Oleanolic Acid bullion, filters, and filtrating is regulated PH between 3-4 with the 6mol/L aqueous hydrochloric acid.In filtrating, add 5L water, place 4 ℃ and spend the night, have a large amount of depositions to separate out.Filter, precipitate is the ursolic acid bullion.
The purifying of Oleanolic Acid:
The above-mentioned Oleanolic Acid bullion 17.9g that obtains is washed with an amount of pure water; Water-insoluble is used the 285ml anhydrous alcohol solution, regulates PH to 11 with 10% aqueous sodium hydroxide solution, adds the 5.7g gac; Placed 50-60 ℃ of stirring in water bath 15 minutes; The filtering gac, filtrating is regulated PH to 1 with the 6mol/L aqueous hydrochloric acid, separates out throw out 15.4g.Throw out is joined 90ml, boiled in 5% aqueous sodium hydroxide solution 30 minutes, filter, insolubles is boiled the pure water washing with 60ml; Filtered while hot must be washed thing 13.1g, will wash thing and use the 120ml anhydrous alcohol solution, adds the 1.2g gac; Placed 50-60 ℃ of stirring in water bath 15 minutes, and filtered, filtrating is regulated PH to 1 with the 6mol/L aqueous hydrochloric acid; Separate out Oleanolic Acid, filter, the zero(ppm) water that boils washing obtains Oleanolic Acid 8.7g.Product is through Spectrum Analysis, its result and Oleanolic Acid conformance to standard.
The purifying of ursolic acid:
The above-mentioned ursolic acid bullion 62g that obtains is washed with an amount of 1% sodium hydroxide solution; Then with an amount of pure water washing; With insolubles 50.7g with an amount of absolute ethyl alcohol recrystallization twice, use again the 200ml volume ratio be 1: 1 the crystallization of acetate ethanol mixing solutions once, finally obtain ursolic acid 29.7g.Product is through Spectrum Analysis, its result and ursolic acid conformance to standard.
Claims (6)
1. the method for coproduce high-purity ursolic acid and Oleanolic Acid from a glossy privet, said glossy privet comprises Leaf of Glossy Privet or Glossy Privet Fruit, this method comprises the following steps, it is characterized in that:
A) glossy privet blade or Glossy Privet Fruit are boiled with buck, filtered while hot, alkali compound is used hot wash, removes content of wax impurity.With the dregs of a decoction that obtain is extracting solution with the pure water, in the boiling water water-bath, extracts, filter, and with the filter residue that filtration obtains, be solvent with the ethanolic soln, the refluxing extraction after-filtration obtains ethanol extract in water-bath;
B) ethanol extract is used activated carbon decolorizing, the filtering gac is regulated ethanol extract pH value to alkalescence; After leaving standstill impurity screening filtrate decompression is concentrated into small volume, filters, leaching thing is the Oleanolic Acid bullion; Filtrating is regulated the pH value to acid, adds isopyknic pure water, places 4 ℃ to spend the night; Leach insolubles, obtain the ursolic acid bullion;
C) the Oleanolic Acid bullion is washed with pure water, insolubles is used anhydrous alcohol solution, and regulating PH is alkalescence, adds activated carbon decolorizing, places and filters; Filtrating transfers to acidity, separates out throw out, and throw out is boiled with buck, filters, and repeats 2-3 time; Insolubles is used boiling pure water, and filtered while hot must be washed thing, will wash thing and use anhydrous alcohol solution, activated carbon decolorizing once more; Filter, filtrating transfers to acidity, separates out throw out, and the zero(ppm) water that boils washing can obtain highly purified Oleanolic Acid;
D) the ursolic acid bullion is washed with 1%NaOH, then washed, use absolute ethyl alcohol recrystallization twice then, use methanol mixing solutions or acetate ethanol mixing solutions recrystallization at last, promptly obtain highly purified ursolic acid with pure water.
2. method according to claim 1; It is characterized in that: buck is 5% sodium hydroxide or potassium hydroxide aqueous solution steps A); The add-on of buck (mL) is 6-8 times with the liquid-solid ratio of Leaf of Glossy Privet that adds or Glossy Privet Fruit quality (g) during removal of impurities, and boiling time was controlled at 20-30 minute;
The add-on of pure water (mL) was 8-12 times with the liquid-solid ratio of Leaf of Glossy Privet or Glossy Privet Fruit quality (g) when water was carried, and extraction time is 1-2 hour in the boiling water water-bath.
The concentration of ethanolic soln is more than 80% during alcohol extracting, and add-on (mL) is 10-15 times with the liquid-solid ratio of Leaf of Glossy Privet or Glossy Privet Fruit quality (g), and each extraction time is 1.5-2 hour, extracts 2-3 time.
3. method according to claim 1 is characterized in that: used accent alkali reagent is 1mol/L sodium hydroxide or potassium hydroxide aqueous solution step B), and PH should be controlled in the 9-11 scope;
Used acid adjustment reagent is the 6mol/L aqueous hydrochloric acid, and PH should be controlled in the 3-4 scope;
Filtrate decompression concentrates volume and should be in the 1/3-1/4 scope that concentrates front volume.
4. method according to claim 1 is characterized in that: used accent alkali reagent is 10% sodium hydroxide or potassium hydroxide aqueous solution among the step C, and PH should be controlled at 11;
Used acid adjustment reagent is the 6mol/L aqueous hydrochloric acid, and PH should be controlled at 1;
The used buck that boils is 5% sodium hydroxide or potassium hydroxide aqueous solution, and boiling time should be controlled at 15-30 minute, and the liquid-solid ratio of institute's consumption (ml) and throw out (g) is 6-8 times.
5. method according to claim 1 is characterized in that: used buck washing lotion is 1% sodium hydroxide or potassium hydroxide aqueous solution among the step D.The methyl alcohol that recrystallization adopted and the volume ratio of alcohol mixed solution are 1: 1, and the volume ratio of ethanol that recrystallization adopted and acetic acid mixing solutions is 1: 1.
6. method according to claim 1, decolouring alcohol extract adding liquid-solid ratio is the 0.1-1.0% gac in each step, whip attachment 15-60 minute, repeat 2-3 time, bleaching temperature is 50-60 ℃.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101768201A (en) * | 2010-02-08 | 2010-07-07 | 苏州宝泽堂医药科技有限公司 | Preparation method of oleanolic acid |
| CN101775060A (en) * | 2010-02-08 | 2010-07-14 | 苏州宝泽堂医药科技有限公司 | Method for separating and purifying ursolic acid from glossy privet leaves |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101768201A (en) * | 2010-02-08 | 2010-07-07 | 苏州宝泽堂医药科技有限公司 | Preparation method of oleanolic acid |
| CN101775060A (en) * | 2010-02-08 | 2010-07-14 | 苏州宝泽堂医药科技有限公司 | Method for separating and purifying ursolic acid from glossy privet leaves |
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Application publication date: 20120523 |