CN103450135A - Method for extracting and purifying Broussoflavonol A - Google Patents
Method for extracting and purifying Broussoflavonol A Download PDFInfo
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- CN103450135A CN103450135A CN2013103499977A CN201310349997A CN103450135A CN 103450135 A CN103450135 A CN 103450135A CN 2013103499977 A CN2013103499977 A CN 2013103499977A CN 201310349997 A CN201310349997 A CN 201310349997A CN 103450135 A CN103450135 A CN 103450135A
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- acetone
- broussoflavone
- alcohol
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- extraction
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a method for extracting and purifying Broussoflavonol A, which comprises the following steps: (1) drying and pulverizing broussonetia papyrifera roots and barks used as raw materials, and extracting oil and fat substances in the broussonetia papyrifera roots and barks by using a supercritical carbon dioxide technology; (2) adding an acetone solution into the residue, and performing ultrasonic recycling extraction; (3) concentrating the extracting solution, loading onto a macroporous resin column, eluting with water and a 40-60% ethanol solution, then eluting with a 85-95% ethanol solution, and collecting the 85-95% ethanol eluate; (4) concentrating the eluate, loading onto a silica gel short wide column, collecting the eluate by taking an ethyl acetate-acetone mixed solution as an eluting agent, concentrating, and standing for crystallization; and (5) separating out crystals, recrystallizing with acetone-n-hexane mixed solvent, filtering out the crystals, and performing freeze-drying to obtain the Broussoflavonol A. The method disclosed by the invention is simple to operate and can realize large-scale production.
Description
Technical field
The invention belongs to natural medicine field, particularly, the present invention relates to a kind of method of extracting purifying broussoflavone alcohol A.
Background technology
Broussoflavone alcohol A(Broussoflavonol A), be a kind of flavonols compound, molecular formula: C
25h
30o
6, molecular weight: 426.51, yellow amorphous powder (acetone-hexane), mp238-240 ℃.Mainly be present in the root skin of Moraceae (Moraceae) paper mulberry.
Modern pharmacological research shows, broussoflavone alcohol A can suppress lipid oxidation, in the mouse brain homogenate, suppresses by Fe
2+the thiobarbital acid reaction caused generates, its IC
50be 2.1 μ M; Suppress rat smooth muscle cells (VSMC) propagation, making stimulates [3H]-thymidine and DNA fusion percentage to reduce to by following three things: foetal calf serum (FCS, 10%), 39.5%; ADP (10 μ M), 67%; 5-HT (10 μ M), 19%; Platelet aggregation-against, the platelet aggregation of inhibition arachidonic acid-induction, its IC
50be 86.7 μ M.
By literature search, there is no the extracting and purifying method report that is applicable to high purity broussoflavone alcohol A.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of method that is applicable to the high extraction purifying broussoflavone alcohol A of large-scale production, product purity.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
(1) take paper mulberry root skin is raw material, carries out drying, pulverizing, adopts the oil substances in super critical CO 2 technology extraction paper mulberry root skin;
(2) residue add 5-10 doubly (V/W) 75-95% acetone soln carry out ultrasonic circulating and extract 1-3 time, 1-2h at every turn, united extraction liquid;
(3) the concentrated rear upper macroporous resin column of extracting solution, first water and 40-60% ethanolic soln wash-out, then use 85-95% ethanolic soln wash-out, collect the 85-95% ethanol eluate;
(4) elutriant is concentrated, upper silica gel short wide column, and the post blade diameter length ratio is 1:4-5, the ethyl acetate-acetone mixing solutions of take is eluent, collects elutriant, concentrated, places crystallization;
(5) separate crystallization, acetone-normal hexane mixed solvent recrystallization, leach crystallization, and lyophilize obtains broussoflavone alcohol A.
In described step (1), the supercritical carbon dioxide extraction temperature is 35-50 ℃, and pressure is 20-32MPa, and extraction time is 0.5-1h.
The optional HPD100 of macroporous resin, ADS-17, D101 or HZ801 type in described step (3).
Ethyl acetate in described step (4)-acetone mixed liquor volume is than being 1:5-15.
In described step (5), acetone-normal hexane mixed solvent volume ratio is 1-1.5:1.
In sum, there is following advantage in the present invention:
(1) adopt supercritical carbon dioxide extraction method to remove oil substances, fast, efficiently.
(2) method that adopts ultrasonic circulating to extract, extraction efficiency is high.
(3) carry out crystallization after employing macroporous resin and silica gel short wide column chromatography,, thus the broussoflavone alcohol A of content more than 98% obtained, and cost is low, good separating effect.
(4) agents useful for same ethyl acetate of the present invention, acetone etc., chloroform, methyl alcohol that alternative common column chromatography adopts, reduce toxicity and pollution.
Below in conjunction with embodiment, the present invention is further elaborated, but the scope of protection of present invention is not limited to the following example.
Embodiment
Embodiment 1:
Get paper mulberry root skin meal 500g, adopt the oil substances in super critical CO 2 technology extraction paper mulberry root skin, extraction temperature is 35 ℃, pressure is 20MPa, extraction time is 1h, residue adds 10 times of (V/W) 90% acetone solns and carries out ultrasonic circulating extraction 1 time, each 1h, united extraction liquid, upper HPD100 type macroporous resin column after concentrated, first water and 40% ethanolic soln wash-out, use again 85% ethanolic soln wash-out, collect 85% ethanol eluate, concentrated, with 100-200 purpose silica gel, by weight 1:1, mix thoroughly, dry in the shade, upper silica gel short wide column, the post blade diameter length ratio is 1:5, ethyl acetate-acetone the mixing solutions of take is eluent, press successively 1:5, 1:10, the 1:15 volume ratio is carried out wash-out, collect the elutriant containing broussoflavone alcohol A, reclaim reagent, gained broussoflavone alcohol A concentrated solution is placed in to crystallizer and within standing 20 hours, makes its crystallization, filter, separate crystallization, adopt acetone-normal hexane (1.5:1) mixed solvent recrystallization, leach crystallization, lyophilize obtains broussoflavone alcohol A1.1A, purity is 97.4%.
Embodiment 2:
Get paper mulberry root skin meal 500g, adopt the oil substances in super critical CO 2 technology extraction paper mulberry root skin, extraction temperature is 40 ℃, pressure is 25MPa, extraction time is 0.5h, residue adds 7 times of (V/W) 80% acetone solns and carries out ultrasonic circulating extraction 2 times, each 1h, united extraction liquid, upper HZ801 type macroporous resin column after concentrated, first water and 45% ethanolic soln wash-out, use again 90% ethanolic soln wash-out, collect 90% ethanol eluate, concentrated, with 100-200 purpose silica gel, by weight 1:1, mix thoroughly, dry in the shade, upper silica gel short wide column, the post blade diameter length ratio is 1:4, ethyl acetate-acetone the mixing solutions of take is eluent, press successively 1:5, 1:9, the 1:13 volume ratio is carried out wash-out, collect the elutriant containing broussoflavone alcohol A, reclaim reagent, gained broussoflavone alcohol A concentrated solution is placed in to crystallizer and within standing 24 hours, makes its crystallization, filter, separate crystallization, adopt acetone-normal hexane (1:1) mixed solvent recrystallization, leach crystallization, lyophilize obtains broussoflavone alcohol A1.3A, purity is 98.2%.
Embodiment 3:
Get paper mulberry root skin meal 500A, adopt the oil substances in super critical CO 2 technology extraction paper mulberry root skin, extraction temperature is 45 ℃, pressure is 28MPa, extraction time is 0.5h, residue adds 10 times of (V/W) 90% acetone solns and carries out ultrasonic circulating extraction 1 time, each 2h, united extraction liquid, upper D101 type macroporous resin column after concentrated, first water and 60% ethanolic soln wash-out, use again 85% ethanolic soln wash-out, collect 85% ethanol eluate, concentrated, with 100-200 purpose silica gel, by weight 1:1, mix thoroughly, dry in the shade, upper silica gel short wide column, the post blade diameter length ratio is 1:5, ethyl acetate-acetone the mixing solutions of take is eluent, press successively 1:5, 1:10, the 1:15 volume ratio is carried out wash-out, collect the elutriant containing broussoflavone alcohol A, reclaim reagent, gained broussoflavone alcohol A concentrated solution is placed in to crystallizer and within standing 12 hours, makes its crystallization, filter, separate crystallization, adopt acetone-normal hexane (1.5:1) mixed solvent recrystallization, leach crystallization, lyophilize obtains broussoflavone alcohol A1.0A, purity is 98.6%.
Embodiment 4:
Get paper mulberry root skin meal 1kA, adopt the oil substances in super critical CO 2 technology extraction paper mulberry root skin, extraction temperature is 50 ℃, pressure is 32MPa, extraction time is 0.5h, residue adds 6 times of (V/W) 70% acetone solns and carries out ultrasonic circulating extraction 3 times, each 1h, united extraction liquid, upper ADS-17 type macroporous resin column after concentrated, first water and 55% ethanolic soln wash-out, use again 95% ethanolic soln wash-out, collect 95% ethanol eluate, concentrated, with 100-200 purpose silica gel, by weight 1:1, mix thoroughly, dry in the shade, upper silica gel short wide column, the post blade diameter length ratio is 1:3, ethyl acetate-acetone the mixing solutions of take is eluent, press successively 1:6, 1:11, the 1:14 volume ratio is carried out wash-out, collect the elutriant containing broussoflavone alcohol A, reclaim reagent, gained broussoflavone alcohol A concentrated solution is placed in to crystallizer and within standing 18 hours, makes its crystallization, filter, separate crystallization, adopt acetone-normal hexane (1:1) mixed solvent recrystallization, leach crystallization, lyophilize obtains broussoflavone alcohol A2.0A, purity is 98.9%.
Embodiment 5:
Get paper mulberry root skin meal 5kA, adopt the oil substances in super critical CO 2 technology extraction paper mulberry root skin, extraction temperature is 38 ℃, pressure is 30MPa, extraction time is 0.5h, residue adds 10 times of (V/W) 80% acetone solns and carries out ultrasonic circulating extraction 3 times, each 2h, united extraction liquid, upper HPD100 type macroporous resin column after concentrated, first water and 50% ethanolic soln wash-out, use again 90% ethanolic soln wash-out, collect 90% ethanol eluate, concentrated, with 100-200 purpose silica gel, by weight 1:1, mix thoroughly, dry in the shade, upper silica gel short wide column, the post blade diameter length ratio is 1:5, ethyl acetate-acetone the mixing solutions of take is eluent, press successively 1:5, 1:11, the 1:15 volume ratio is carried out wash-out, collect the elutriant containing broussoflavone alcohol A, reclaim reagent, gained broussoflavone alcohol A concentrated solution is placed in to crystallizer and within standing 12 hours, makes its crystallization, filter, separate crystallization, adopt acetone-normal hexane (1:1) mixed solvent recrystallization, leach crystallization, lyophilize obtains broussoflavone alcohol A8.3A, purity is 99.1%.
Claims (5)
1. a method of extracting purifying broussoflavone alcohol A is characterized in that being undertaken by following processing step:
(1) take paper mulberry root skin is raw material, carries out drying, pulverizing, adopts the oil substances in super critical CO 2 technology extraction paper mulberry root skin;
(2) residue add 5-10 doubly (V/W) 75-95% acetone soln carry out ultrasonic circulating and extract 1-3 time, 1-2h at every turn, united extraction liquid;
(3) the concentrated rear upper macroporous resin column of extracting solution, first water and 40-60% ethanolic soln wash-out, then use 85-95% ethanolic soln wash-out, collect the 85-95% ethanol eluate;
(4) elutriant is concentrated, upper silica gel short wide column, and the post blade diameter length ratio is 1:4-5, the ethyl acetate-acetone mixing solutions of take is eluent, collects elutriant, concentrated, places crystallization;
(5) separate crystallization, acetone-normal hexane mixed solvent recrystallization, leach crystallization, and lyophilize obtains broussoflavone alcohol A.
2. a kind of method of extracting purifying broussoflavone alcohol A as claimed in claim 1, is characterized in that, in described step (1), the supercritical carbon dioxide extraction temperature is 35-50 ℃, and pressure is 20-32MPa, and extraction time is 0.5-1h.
3. a kind of method of extracting purifying broussoflavone alcohol A as claimed in claim 1, is characterized in that the optional HPD100 of macroporous resin, ADS-17, D101 or HZ801 type in described step (3).
4. a kind of method of extracting purifying broussoflavone alcohol A as claimed in claim 1, is characterized in that, ethyl acetate in described step (4)-acetone mixed liquor volume is than being 1:5-15.
5. a kind of method of extracting purifying broussoflavone alcohol A as claimed in claim 1, is characterized in that, in described step (5), acetone-normal hexane mixed solvent volume ratio is 1-1.5:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN2013103499977A CN103450135A (en) | 2013-08-13 | 2013-08-13 | Method for extracting and purifying Broussoflavonol A |
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|---|---|---|---|
| CN2013103499977A CN103450135A (en) | 2013-08-13 | 2013-08-13 | Method for extracting and purifying Broussoflavonol A |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105198945A (en) * | 2015-10-30 | 2015-12-30 | 河南中医学院 | Novel broussonetia papyrifera lactone B with antitumor effect as well as preparation method and application thereof |
| EP3120712A1 (en) | 2015-07-22 | 2017-01-25 | Evonik Degussa GmbH | Method for improved extraction of juniper berries, rose hips, sea buckthorn berries, sorbus |
| US10144904B2 (en) | 2015-12-04 | 2018-12-04 | Evonik Degussa Gmbh | Process for extraction of aroma chemicals from fat-containing and/or aqueous liquid phases |
-
2013
- 2013-08-13 CN CN2013103499977A patent/CN103450135A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3120712A1 (en) | 2015-07-22 | 2017-01-25 | Evonik Degussa GmbH | Method for improved extraction of juniper berries, rose hips, sea buckthorn berries, sorbus |
| CN105198945A (en) * | 2015-10-30 | 2015-12-30 | 河南中医学院 | Novel broussonetia papyrifera lactone B with antitumor effect as well as preparation method and application thereof |
| US10144904B2 (en) | 2015-12-04 | 2018-12-04 | Evonik Degussa Gmbh | Process for extraction of aroma chemicals from fat-containing and/or aqueous liquid phases |
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Application publication date: 20131218 |