CN102827241A - Method for extracting betulinic acid from persimmon leaves - Google Patents
Method for extracting betulinic acid from persimmon leaves Download PDFInfo
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- CN102827241A CN102827241A CN 201210350319 CN201210350319A CN102827241A CN 102827241 A CN102827241 A CN 102827241A CN 201210350319 CN201210350319 CN 201210350319 CN 201210350319 A CN201210350319 A CN 201210350319A CN 102827241 A CN102827241 A CN 102827241A
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- betulinic acid
- extracting
- persimmon leaf
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- enzymolysis
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Abstract
The invention discloses a method for extracting betulinic acid from persimmon leaves. The method comprises the steps that dry persimmon leaves are taken to be ground, added with water to mix and wet, added with bio-enzyme for natural enzymolysis for 4-7 days, the enzymolysis raw material is added with 70-90% ethanol and extracted under refluxing for 1-3 times, the extract is concentrated under reduced pressure until the alcohol concentration is 50-60%, and stood for crystallization, crystal substances are dissolved with 5-7 times of amount of alkaline ethanol solution, undissolved substances are filtered out, filtrate is regulated through hydrochloric acid or sulfuric acid solution to be neutral and centrifuged, crude extract is dissolved with ethanol for recrystallizing, and the betulinic acid is obtained after drying. By adopting the method for extracting the betulinic acid, the raw material resources are abundant, the method is simple to operate, and the industrialization production is benefited.
Description
Technical field
The invention belongs to biological technical field, particularly relate to a kind of method of from the persimmon leaf, extracting Betulinic acid.
Background technology
Betulinic acid is betulinic ncid again, and CAS number is 472-15-1, and molecular formula is C
30H
48O
3, molecular weight is 456.71, molecular structural formula is:
Betulinic acid is a pentacyclic triterpenoid, and water insoluble and sherwood oil is slightly soluble in methyl alcohol, ethanol, acetone, is soluble in THF, pyridine.Betulinic acid can optionally kill human MC and not kill and wound healthy cell, and the HIV-1 type is infected also has restraining effect.Recent in addition research shows that Betulinic acid also has restraining effect to malignant cells such as brain tumor, PNET, white blood disease.
The present main source of Betulinic acid has three kinds of approach: first from natural phant extraction separation obtain; Second is precursor with the trochol, obtains Betulinic acid through organic synthesis; The 3rd, be precursor with the trochol, generate Betulinic acid through microbial transformation.The back all is that raw material extracts with the Cortex Betulae Pendulae for two kinds, because the Cortex Betulae Pendulae raw material is limited, output is restricted always.Discover and contain more Betulinic acid in the persimmon leaf, generally at 0.3-0.5%, and China persimmon distributes extensively, plant morely, and the persimmon leaf generally acts as a fuel and fertilizer is handled, and is raw material extraction Betulinic acid with the persimmon leaf, the shortage that can solve the Cortex Betulae Pendulae raw material.
The technology of existing extraction Betulinic acid is more.(application number: 03131059.1) " a kind of method of extracting Betulinic acid " disclosed method is for being that raw material pulverizing is a meal with the Spina Date Seed like patent; Remove degrease with sherwood oil; Extract with methylene chloride reflux again; Reclaim solvent, the thick paste of gained through complexing, separate steps such as complexing, recrystallization, post separation, obtain the different purity Betulinic acid.(application number: disclosed method is to be raw material with the Fructus Camptothecae Acuminatae to patent 200710031330.7) " from Fructus Camptothecae Acuminatae, to extract effective constituents such as Betulinic acid with cleaning method ", utilizes the method for polyamide column chromatography and ethanol elution for another example.
As stated, the existing Betulinic acid method of producing is comparatively loaded down with trivial details, and reagent type is more, and especially macroporous resin and polyamide resin are handled and be difficult to operation, and eluting solvent is more.
Summary of the invention
The objective of the invention is to overcome the defective and the deficiency of prior art, a kind of method of from the persimmon leaf, extracting Betulinic acid is provided.This method is simple to operate, and raw material is easy to get.
The objective of the invention is to realize through following technical scheme:
Get dry persimmon leaf and pulverize, water is mixed wet, adds the enzyme natural enzymolysis 4-7 days; The ethanolic soln refluxing extraction of enzymolysis raw material adding 70-90% 1-3 time, extracting solution is evaporated to determining alcohol 50-60%, places crystallization; Crystallisate is doubly measured the alkaline ethanol solution dissolving with 5-7, leaches insolubles, regulates neutral with hydrochloric acid or sulphuric acid soln; Centrifugal, crude extract is used the dissolve with ethanol recrystallization again, is drying to obtain Betulinic acid.
A kind of or arbitrary combination in the optional cellulase of described enzyme, glycase and the polygalacturonase, consumption are the 1-3 ‰ of material quantity.
Described alkaline ethanol solution is the 70-90% ethanolic soln of pH10-14, optional sodium hydroxide of alkali or Pottasium Hydroxide.
Advantage of the present invention is:
1. raw material resources are abundant, and cost is low;
2. method adopts enzymolysis process degraded raw material, can shorten extracting cycle, improves yield, has also reduced the impurity stripping quantity;
3. whole technological operation is simple, is easy to suitability for industrialized production.
Embodiment:
To combine embodiment to further specify the present invention below.
Embodiment 1:
Get dry persimmon leaf and pulverize, claim that 50kg adds 10kg water and mixes wetly, add cellulase 150g; Mix thoroughly, natural enzymolysis 4 days, the enzymolysis raw material drops into extractor; The ethanolic soln refluxing extraction of adding 250kg70% 3 times is evaporated to determining alcohol about 55% behind the extracting liquid filtering, emitted crystallization 10 hours; Centrifugal, get crystallisate 800g.Get the 70% dissolve with ethanol solution crystallisate of 4kgpH13, fully stir, leach insolubles, regulate neutrality with hydrochloric acid soln, centrifugal, get crude extract 327g.Crude extract is used 90% alcohol reflux dissolving-recrystallization 3 times again, and drying gets white crystal Betulinic acid 83g, detects content 96.7% through performance liquid.
Embodiment 2:
Get dry persimmon leaf and pulverize, claim that 50kg adds 10kg water and mixes wetly, add cellulase and glycase 150g; Mix thoroughly; Natural enzymolysis 7 days, the enzymolysis raw material drops into extractor, the ethanolic soln refluxing extraction of adding 350kg90% 2 times; Be evaporated to determining alcohol about 60% behind the extracting liquid filtering and emitted crystallization 8 hours, the centrifugal crystallisate 721g that gets.Get the 90% dissolve with ethanol solution crystallisate of 4kgpH10, fully stir, leach insolubles, regulate neutrality with sulphuric acid soln, centrifugal, get crude extract 285g.Crude extract is used 95% alcohol reflux dissolving-recrystallization 2 times again, and drying gets white crystal Betulinic acid 91g, detects content 97.3% through performance liquid.
Embodiment 3:
Get dry persimmon leaf and pulverize, claim that 50kg adds 10kg water and mixes wetly, add polygalacturonase and amylase 8 0g; Mix thoroughly, natural enzymolysis 7 days, the enzymolysis raw material drops into extractor; The ethanolic soln refluxing extraction of adding 300kg80% 2 times is evaporated to determining alcohol about 55% behind the extracting liquid filtering, emitted crystallization 12 hours; Centrifugal, get crystallisate 815g.Get the 85% dissolve with ethanol solution crystallisate of 5kgpH10, fully stir, leach insolubles, regulate neutrality with sulphuric acid soln, centrifugal, get crude extract 272g.Crude extract is used 95% alcohol reflux dissolving-recrystallization 3 times again, and drying gets white crystal Betulinic acid 75g, detects content 95.1% through performance liquid.
Claims (3)
1. method of from the persimmon leaf, extracting Betulinic acid, it is characterized in that: get dry persimmon leaf and pulverize, water is mixed wet, adds the enzyme natural enzymolysis 4-7 days; The ethanolic soln refluxing extraction of enzymolysis raw material adding 70-90% 1-3 time, extracting solution is evaporated to determining alcohol 50-60%, places crystallization; Crystallisate is doubly measured the alkaline ethanol solution dissolving with 5-7, leaches insolubles, and filtrating is regulated neutral with hydrochloric acid or sulphuric acid soln; Centrifugal, crude extract is used the dissolve with ethanol recrystallization again, is drying to obtain Betulinic acid.
2. the method for from the persimmon leaf, extracting Betulinic acid according to claim 1 is characterized in that a kind of or arbitrary combination in the optional cellulase of described enzyme, glycase and the polygalacturonase, and consumption is the 1-3 ‰ of material quantity.
3. the method for from the persimmon leaf, extracting Betulinic acid according to claim 1 is characterized in that described alkaline ethanol solution is the 70-90% ethanolic soln of pH10-14, optional sodium hydroxide of alkali or Pottasium Hydroxide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210350319 CN102827241A (en) | 2012-09-20 | 2012-09-20 | Method for extracting betulinic acid from persimmon leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210350319 CN102827241A (en) | 2012-09-20 | 2012-09-20 | Method for extracting betulinic acid from persimmon leaves |
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|---|---|
| CN102827241A true CN102827241A (en) | 2012-12-19 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 201210350319 Pending CN102827241A (en) | 2012-09-20 | 2012-09-20 | Method for extracting betulinic acid from persimmon leaves |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109666058A (en) * | 2018-12-25 | 2019-04-23 | 东南大学 | A kind of method that betulic acid extraction is isolated |
| CN112521441A (en) * | 2020-12-29 | 2021-03-19 | 江西省科学院应用化学研究所 | Method for separating and purifying betulinic acid from lotus seedpod shells |
-
2012
- 2012-09-20 CN CN 201210350319 patent/CN102827241A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109666058A (en) * | 2018-12-25 | 2019-04-23 | 东南大学 | A kind of method that betulic acid extraction is isolated |
| CN112521441A (en) * | 2020-12-29 | 2021-03-19 | 江西省科学院应用化学研究所 | Method for separating and purifying betulinic acid from lotus seedpod shells |
| CN112521441B (en) * | 2020-12-29 | 2021-09-28 | 江西省科学院应用化学研究所 | Method for separating and purifying betulinic acid from lotus seedpod shells |
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| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20121219 |