CN103254210A - Method for preparing castalagin - Google Patents
Method for preparing castalagin Download PDFInfo
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- CN103254210A CN103254210A CN2013101959478A CN201310195947A CN103254210A CN 103254210 A CN103254210 A CN 103254210A CN 2013101959478 A CN2013101959478 A CN 2013101959478A CN 201310195947 A CN201310195947 A CN 201310195947A CN 103254210 A CN103254210 A CN 103254210A
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- castalagin
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- extraction
- extract
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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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Abstract
The invention discloses a method for preparing castalagin. According to the method, anogeissus barks are used as raw materials. The technical flow of the method mainly comprises four steps of: (1) extracting fat-type substances from anogeissus bark medicinal materials by supercritical carbon dioxide fluid, and performing ultrasonic extraction to obtain an extract; (2) concentrating the extract at a low temperature, adding gelatine solution into the extract, performing centrifugation, dissolving precipitates by acetone aqueous solution, and filtering out gel to obtain preliminarily purified liquid; (3) purifying the preliminarily purified liquid by macroporous resin, eluting the preliminarily purified liquid by water and ethanol water, and concentrating the eluant; and (4) carrying out preparative liquid chromatography separation on the concentrated liquid to obtain castalagin, thus obtaining the castalagin. The preparation method is executed at the temperature being less than 50 DEG C; the chemical and structural changes of active components caused by overhigh temperature in the conventional thermal backflow extraction process are avoided; therefore, by the method, castalagin with higher purity can be obtained; castalagin content of a product is over 95 percent; the method is short in extraction period, safe and suitable for industrial production; and the method lays a foundation for deep research on the pharmacological activity and pharmaceutic preparations of tannin type components.
Description
Technical field
The present invention relates to a kind of preparation method of active ingredient of Chinese herbs, specifically a kind of method that from the green wooden stem follicarpium of elm is real, prepares castalagin.
Background technology
Castalagin (Castalagin) is from the green wooden mutation of Combretum Racemosum (Combretaceae) elm
Anogeissus acuminata(Roxb.ex DC.) Guill.﹠amp; Pers.var.
LanceolataThe isolated a kind of tannin compounds of the stem of Wall.ex C.B.Clarke is off-white powder, and molecular formula is C
19H
18O
4, molecular weight is 934.34, structural formula is
Pharmacological research shows that castalagin has antihypertensive active, and quiet notes this product reduces the blood pressure of spontaneous hypertensive rat, and this is owing to suppress due to orthosympathetic activity and the direct vasodilation; Having cytotoxic activity, is 0.79 μ g/ml to the ED50 of melanoma RPMI-7951.
At present, domestic research to castalagin is less, still is in the starting stage.Therefore, work out a kind of method for preparing castalagin and be very important, have very important significance for the comprehensive utilization tool that enlarges medicine source and the green wooden stem skin of elm.
Summary of the invention
Technical problem to be solved by this invention provides a kind of simple to operate, the castalagin preparation method that product purity is higher.
For solving the problems of the technologies described above, the present invention adopts following technical proposal:
A kind of preparation method of castalagin is characterized in that may further comprise the steps:
(1) oil substances in the green wooden stem skin medicinal material of supercritical carbon dioxide extraction elm, residue obtain extracting solution after with ethanol ultrasonic extraction;
(2) extracting solution concentrates, and adds gelatin solution, and centrifugal, throw out dissolves with aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid;
(3) go up macroporous resin purification, water and aqueous ethanolic solution wash-out, elutriant concentrates;
(4) concentrated solution separates through preparative liquid chromatography, and lyophilize namely gets castalagin.
The supercritical extraction temperature is 35-50 ℃ described in the step (1), and pressure is 25-30MPa, and the time is 30-60min.
Ethanol described in the step (1) is the ethanolic soln solution of 60-80%, and consumption is that 8-10 doubly measures (V/W), and ultrasonic frequency is 45-65KHz, extracts 2-3 time, each 0.5-1h.
Gelatin solution concentration is 4.5-6% described in the step (2), and aqueous acetone solution concentration is 50-70%.
The optional HPD-400 of macroporous resin or ADS-17 type described in the step (3), eluent is 70% aqueous ethanolic solution.
Preparative liquid chromatography described in the step (4) separates, and is moving phase with the methanol-water, and the methyl alcohol volume percent is 68%, and flow velocity is 25-50ml/min.
In sum, there is following advantage in the present invention: the method for this process using ultrasonic extraction is extracted castalagin, has improved extraction efficiency; The ethanolic soln of 60-80% has reduced the stripping of polysaccharide as extracting solvent; Adopt macroporous resin to separate castalagin, cost is low, good separating effect; The preparative liquid chromatography separation and purification, it is simple to have production process, is easy to realize advantages such as industrialization.
Below in conjunction with embodiment the present invention is further elaborated, but the scope of protection of present invention is not limited to following embodiment.
Embodiment:
Embodiment 1:
The green wooden stem skin of elm is pulverized, and gets 1kg, puts into supercritical extraction unit, be 35 ℃ in temperature, pressure is to extract 30min under the 25MPa, and the ethanolic soln that residue adds 8L80% carries out supersound extraction, ultrasonic frequency is 45KHz, extracts 3 times, each 1h, filter united extraction liquid, cryoconcentration, adding concentration is 3% gelatin solution, is stirred to no longer to produce precipitation, centrifugal, throw out dissolves with 50% aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid, last 1LHPD-400 macroporous resin column after absorption is finished, is used earlier the 4L water elution, use 4L70% ethanolic soln wash-out again, the control flow velocity is 3.5ml/min, collects the elutriant that is rich in castalagin, concentrates, concentrated solution is added methyl alcohol be mixed with settled solution, separate with preparative high performance liquid chromatography, preparative column is column packing with C18, is moving phase with 68% methanol aqueous solution, the control flow velocity is 25ml/min, on-line ultraviolet detects, and the detection wavelength is 229nm, collects the target flow point, lyophilize namely gets castalagin 1.68g, and content is 96.4%.
Embodiment 2:
The green wooden stem skin of elm is pulverized, and gets 1kg, puts into supercritical extraction unit, be 45 ℃ in temperature, pressure is to extract 50min under the 30MPa, and the ethanolic soln that residue adds 10L70% carries out supersound extraction, ultrasonic frequency is 65KHz, extracts 2 times, each 1h, filter united extraction liquid, cryoconcentration, adding concentration is 6% gelatin solution, is stirred to no longer to produce precipitation, centrifugal, throw out dissolves with 70% aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid, last 1LHPD-400 macroporous resin column after absorption is finished, is used earlier the 5L water elution, use 5L70% ethanolic soln wash-out again, the control flow velocity is 4.5ml/min, collects the elutriant that is rich in castalagin, concentrates, concentrated solution is added methyl alcohol be mixed with settled solution, separate with preparative high performance liquid chromatography, preparative column is column packing with C18, is moving phase with 68% methanol aqueous solution, the control flow velocity is 35ml/min, on-line ultraviolet detects, and the detection wavelength is 229nm, collects the target flow point, lyophilize namely gets castalagin 1.97g, and content is 95.1%.
Embodiment 3:
The green wooden stem skin of elm is pulverized, and gets 1kg, puts into supercritical extraction unit, be 40 ℃ in temperature, pressure is to extract 30min under the 30MPa, and the ethanolic soln that residue adds 10L70% carries out supersound extraction, ultrasonic frequency is 50KHz, extracts 2 times, each 0.5h, filter united extraction liquid, cryoconcentration, adding concentration is 5% gelatin solution, is stirred to no longer to produce precipitation, centrifugal, throw out dissolves with 70% aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid, last 1LADS-17 macroporous resin column after absorption is finished, is used earlier the 3L water elution, use 6L70% ethanolic soln wash-out again, the control flow velocity is 3.5ml/min, collects the elutriant that is rich in castalagin, concentrates, concentrated solution is added methyl alcohol be mixed with settled solution, separate with preparative high performance liquid chromatography, preparative column is column packing with C18, is moving phase with 68% methanol aqueous solution, the control flow velocity is 50ml/min, on-line ultraviolet detects, and the detection wavelength is 229nm, collects the target flow point, lyophilize namely gets castalagin 1.42g, and content is 97.5%.
Embodiment 4:
The green wooden stem skin of elm is pulverized, and gets 1kg, puts into supercritical extraction unit, be 50 ℃ in temperature, pressure is to extract 50min under the 30MPa, and the ethanolic soln that residue adds 9L80% carries out supersound extraction, ultrasonic frequency is 55KHz, extracts 3 times, each 0.5h, filter united extraction liquid, cryoconcentration, adding concentration is 5% gelatin solution, is stirred to no longer to produce precipitation, centrifugal, throw out dissolves with 60% aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid, last 1LHPD-400 macroporous resin column after absorption is finished, is used earlier the 6L water elution, use 3L70% ethanolic soln wash-out again, the control flow velocity is 4ml/min, collects the elutriant that is rich in castalagin, concentrates, concentrated solution is added methyl alcohol be mixed with settled solution, separate with preparative high performance liquid chromatography, preparative column is column packing with C18, is moving phase with 68% methanol aqueous solution, the control flow velocity is 45ml/min, on-line ultraviolet detects, and the detection wavelength is 229nm, collects the target flow point, lyophilize namely gets castalagin 1.60g, and content is 96.2%.
Embodiment 5:
The green wooden stem skin of elm is pulverized, and gets 1kg, puts into supercritical extraction unit, be 50 ℃ in temperature, pressure is to extract 60min under the 28MPa, and the ethanolic soln that residue adds 8L75% carries out supersound extraction, ultrasonic frequency is 60KHz, extracts 2 times, each 1h, filter united extraction liquid, cryoconcentration, adding concentration is 6% gelatin solution, is stirred to no longer to produce precipitation, centrifugal, throw out dissolves with 70% aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid, last 1LHPD-400 macroporous resin column after absorption is finished, is used earlier the 3L water elution, use 4L70% ethanolic soln wash-out again, the control flow velocity is 2.5ml/min, collects the elutriant that is rich in castalagin, concentrates, concentrated solution is added methyl alcohol be mixed with settled solution, separate with preparative high performance liquid chromatography, preparative column is column packing with C18, is moving phase with 68% methanol aqueous solution, the control flow velocity is 40ml/min, on-line ultraviolet detects, and the detection wavelength is 229nm, collects the target flow point, lyophilize namely gets castalagin 1.87g, and content is 95.4%.
Claims (6)
1. the preparation method of a castalagin is characterized in that may further comprise the steps:
(1) oil substances in the green wooden stem skin medicinal material of supercritical carbon dioxide extraction elm, residue obtain extracting solution after with ethanol ultrasonic extraction;
(2) extracting solution concentrates, and adds gelatin solution, and centrifugal, throw out dissolves with aqueous acetone solution, and the filtering gelatin obtains preliminary purification liquid;
(3) go up macroporous resin purification, water and aqueous ethanolic solution wash-out, elutriant concentrates;
(4) concentrated solution separates through preparative liquid chromatography, and lyophilize namely gets castalagin.
2. preparation method as claimed in claim 1 is characterized in that the supercritical extraction temperature is 35-50 ℃ described in the step (1), and pressure is 25-30MPa, and the time is 30-60min.
3. preparation method as claimed in claim 1 is characterized in that ethanol described in the step (1) is the ethanolic soln solution of 60-80%, and consumption is that 8-10 doubly measures (V/W), and ultrasonic frequency is 45-65KHz, extracts 2-3 time, each 0.5-1h.
4. preparation method as claimed in claim 1 is characterized in that gelatin solution concentration is 4.5-6% described in the step (2), and aqueous acetone solution concentration is 50-70%.
5. preparation method as claimed in claim 1 is characterized in that the optional HPD-400 of macroporous resin or ADS-17 type described in the step (3), and eluent is 70% aqueous ethanolic solution.
6. preparation method as claimed in claim 1 is characterized in that preparative liquid chromatography described in the step (4) separates, and is moving phase with the methanol-water, and the methyl alcohol volume percent is 68%, and flow velocity is 25-50ml/min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013101959478A CN103254210A (en) | 2013-05-24 | 2013-05-24 | Method for preparing castalagin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013101959478A CN103254210A (en) | 2013-05-24 | 2013-05-24 | Method for preparing castalagin |
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| CN103254210A true CN103254210A (en) | 2013-08-21 |
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|---|---|---|---|
| CN2013101959478A Pending CN103254210A (en) | 2013-05-24 | 2013-05-24 | Method for preparing castalagin |
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| CN (1) | CN103254210A (en) |
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2013
- 2013-05-24 CN CN2013101959478A patent/CN103254210A/en active Pending
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Application publication date: 20130821 |