CN102942456B - Process for extracting oxyresveratrol from mulberry twigs - Google Patents
Process for extracting oxyresveratrol from mulberry twigs Download PDFInfo
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- CN102942456B CN102942456B CN201210487502.2A CN201210487502A CN102942456B CN 102942456 B CN102942456 B CN 102942456B CN 201210487502 A CN201210487502 A CN 201210487502A CN 102942456 B CN102942456 B CN 102942456B
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- elution
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- oxyresveratrol
- resvertrol
- oxidation
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Abstract
The invention relates to a process for extracting oxyresveratrol from mulberry twigs which serve as raw materials. The process comprises the steps of (1) extracting mulberry twigs by using an alcohol-water solution to obtain crude extracts; (2) conducting degreasing treatment on crude extracts to obtain semi-finished products; (3) dissolving semi-finished products with an alcohol-water solution to produce a loading solution with the turbidity lower than 0.5 degree, using a macroporous resin D101 separation column, conducting wet method loading, standing till resin reaches adsorption equilibrium, conducting gradient elution or isocratic sectional elution with the alcohol-water solution serving as an elution solution, collecting the elution solution with the oxyresveratrol mass fraction larger than 40%, combining, concentrating, and drying to obtain crude products; and (4) repeating purification in Step (3) for crude products, collecting the elution solution with the oxyresveratrol mass fraction larger than or equal to 96.97%, combining, concentrating, and drying to obtain the oxyresveratrol. By the aid of the process, the oxyresveratrol can be extracted from mulberry twigs efficiently, and the process is simple to operate, low in cost and applicable to large-scale production.
Description
Technical field
A kind of method of extracting oxidation resvertrol from ramulus mori of the present invention.
Background technology
Oxidation resvertrol is a kind of iridoid glucoside; the biological activity with multiple beneficial: tyrosinase inhibitory activity, fruit and vegetables corrosion protection anti-microbial activity, anti-herpes simplex virus are active as having, anti-varicella zoster virus activity and AIDS virus resisting isoreactivity also has the effects such as anti-inflammation detumescence, anti-oxidant, anti-apoptotic and neuroprotective simultaneously.
Ramulus mori is the spray of moraceae plants mulberry, and ramulus mori accounts for 64% left and right that amount of dry matter is produced in mulberry field per year, is that in silkworm and mulberry resource, biomass occupies the material that ratio is the highest.Yet in traditional silkworm and mulberry are produced, small part is as fuel, most of mulberry branch goes out of use, and not only its resource value does not obtain fine exploitation, has also polluted environment.
The oxidation resvertrol (be generally 0.01 ‰～1 ‰, only a few kind can reach 1 ‰～4 ‰) known, ramulus mori contains different content with the difference in the place of production.Provide the method for high efficiency extraction oxidation resvertrol from ramulus mori to there is important economy and environment benefit.
Summary of the invention
Technical problem to be solved by this invention be to provide a kind of from ramulus mori the method for high efficiency extraction oxidation resvertrol.
For solving the problems of the technologies described above, the present invention takes following technical scheme:
From ramulus mori, extract a method for oxidation resvertrol, it take ramulus mori as raw material, comprises the following steps of carrying out successively:
(1), with the alcohol solution that alcohol volume content is 65wt% ~ 75wt%, ramulus mori is extracted, concentrated extracting solution, dry, obtain crude extract;
(2), with sherwood oil, step (1) gained crude extract is carried out to skimming treatment, use afterwards ethyl acetate repeatedly to extract, combined ethyl acetate extraction liquid, vacuum decompression reclaims solvent, obtains flow-like material, and 40 ℃ of following vacuum-dryings obtain work in-process;
(3), step (2) gained work in-process are dissolved with ethanol-water solution, make turbidity lower than the loading solution of 0.5 °, adopt macroporous resin D101 separator column, after wet method loading, standing, reach adsorption equilibrium to resin, take ethanol-water solution as elutriant, carry out gradient elution or etc. degree stepwise elution, collect the elutriant that oxidation resvertrol massfraction is greater than 40%, merge, concentrated, dry, obtain crude product;
(4) purification, to step (3) gained crude product repeating step (3), collects oxidation resvertrol massfraction and is more than or equal to 96.97% elutriant, merges, concentrated, dry, obtains described oxidation resvertrol.
Preferably, the described ramulus mori as raw material is dry Mulberry Twig.Size of powder particles can be for example 30 ~ 100 orders.
Preferably, in step (1), alcohol can be that methyl alcohol, ethanol etc. are usually used in the alcohol of plant extract, but is preferably ethanol.Described extraction can be carried out at normal temperatures.
Preferably, in step (2), described vacuum drying temperature is 25 ℃ ~ 30 ℃.
According to a concrete and preferred aspect of the present invention, in step (3), carry out gradient elution.
According to another concrete and preferred aspect of the present invention, in step (4), carry out gradient elution, and be to be more than or equal to 96.97% elutriant merging by the oxidation resvertrol massfraction of collection, concentrated, dry.
According to another concrete and preferred aspect of the present invention, in step (3) and (4), stepwise elution such as degree of grade, 10% the concentration difference of take is a spacer, the elution volume of every section is 2 ~ 3 column volumes, eluent flow rate be 1 ~ 2 column volume/hour.
In step (3), described standingly at room temperature carry out, the standing time is 0.5 ~ 1h.
According to the present invention, in step (1), the consumption of alcohol solution is generally 10 ~ 15 times (V/G) of ramulus mori.
Due to the enforcement of above technical scheme, the present invention compared with prior art tool has the following advantages:
(1) preparation process is simple, easy handling;
(2) with low cost, be applicable to scale operation, there is application potential.
Accompanying drawing explanation
Fig. 1 is the HPLC spectrogram of embodiment 1 gained oxidation resvertrol.
Embodiment
Below in conjunction with specific embodiment, the present invention will be further described in detail, but the invention is not restricted to following examples.
The content assaying method that is oxidized resvertrol in following examples all adopts LC-MS.Instrument: Shimadzu LCMS-2010 HPLC-MS instrument.Comprise 2 LC-10ADvp pumps, FCV-10ALvp quarternary low pressure gradient stream valve, DGU-14AM line vacuum de-aerator, CTO-10Avp column oven, 7725i sampling valve (Rheodyne, USA), SPD-M10Avp diode-array detector, mass spectrum is LCMS-2010 single-stage quadrupole, electron spray ionisation (ESI) ion source, and data acquisition and processing (DAP) is completed by LCMS solution 2.02 softwares.Chromatographic column is Thermo Hypersil-Keystone (Bellefonte, PA, USA) C18 (5 μ m, 150 * 2.1 mm), 40 ℃ of column temperatures.Moving phase is used front through 0.22 μ m membrane filtration ultrasonic degas, and flow velocity is 0.2 mL min-1.Electron spray ionisation (ESI), ion polarity: positive ion (+); Full ion scan (SCAN), sweep limit: 50-1000 (m/z); Probe (probe) temperature: 250 ℃ (ESI); Bent type Desolventizing apparatus (CDL) temperature: 250 ℃ (ESI+); 200 ℃ of heat block (block) temperature; Bent type Desolventizing apparatus (CDL) voltage :-45 V(ESI+); Probe (probe) voltage: 4.5 kV(ESI+); Detect voltage 1.5 kV; Q-array voltage: DC 0V, RF 150V; Atomizing gas flow velocity 2.5 L min-1(ESI).Uv scan scope is 200 to 300 nm.
Embodiment 1
From ramulus mori, extract a method for oxidation resvertrol, it comprises the following step carrying out successively:
(1), ramulus mori (Hunan, the old branch of Huang Lutou, oxidation resvertrol content 1.1 ‰) (moisture content 10%) dried at shady and cool place, take the powder 500kg pulverizing and sieving after (40 order), 70% lixiviate of ethanol-water solution room temperature 3 (4000L * 1,2500L * 2,12hr * 3, every 2hr solution circulated/stirring 10min), merge 3 times ethanol extract, 55 ℃ of vacuum-concentrcteds are to 100L fluid.
(2), by step (1) gained 100L fluid with after petroleum ether degreasing, 3 times (100L * 3) of ethyl acetate extraction, merge three times acetic acid ethyl acetate extract, vacuum decompression reclaims solvent, obtain flow-like material, vacuum-drying at 30 ℃, obtains pressed powder 4.7 kg, and surveying its oxidation resvertrol content is 11.3%.
(3), step (2) gained pressed powder 4.7 kg are added to 30% aqueous ethanolic solution of 35 ℃, make the loading solution of clarification or slightly muddy (turbidity is lower than 0.5 °), adopting macroporous resin D101 is separating medium, after wet method loading, place for some time (approximately 0.5 ~ 1 hour), so that resin reaches adsorption equilibrium to chemical composition; Take ethanol-water solution as elutriant, collect the elutriant that oxidation resvertrol massfraction is greater than 40%, merge, concentrated, dry, obtain the pulverous crude product of 810g.
(4), to step (3) crude product, the purification of repeating step (3), collecting oxidation resvertrol massfraction is to be more than or equal to 96.97% elutriant, merge, concentrated, vacuum-drying 72 hours, must be oxidized 510 grams, resvertrol crystal, through HPLC, detect (going out peak figure referring to Fig. 1), oxidation resvertrol content is 96.97%.
Embodiment 2
A kind of method of extracting oxidation resvertrol from ramulus mori, it is substantially with embodiment 1, different is, step (3) and (4) wash-out adopt the degree stepwise elution such as is, 10% the concentration difference of take is a spacer, the elution volume of every section is 2 column volumes, eluent flow rate be 1 column volume/hour.Finally obtain being oxidized 510 grams, resvertrol crystal, through HPLC, detect, oxidation resvertrol content is 96.97%.
Above the present invention is described in detail; its object is to allow the personage who is familiar with this art can understand content of the present invention and be implemented; can not limit the scope of the invention with this; the equivalence that all spirit according to the present invention are done changes or modifies, and all should be encompassed in protection scope of the present invention.
Claims (5)
1. from ramulus mori, extract a method for oxidation resvertrol, it is characterized in that: take ramulus mori as raw material, comprise the following steps of carrying out successively:
(1), with the alcohol solution that alcohol volume content is 65%～75%, ramulus mori is extracted, concentrated extracting solution, dry, obtain crude extract, described alcohol is ethanol, described extraction is carried out at normal temperatures;
(2), with sherwood oil, step (1) gained crude extract is carried out to skimming treatment, use afterwards ethyl acetate repeatedly to extract, combined ethyl acetate extraction liquid, vacuum decompression reclaims solvent, obtains flow-like material, and 40 ℃ of following vacuum-dryings obtain work in-process;
(3), step (2) gained work in-process are dissolved with ethanol-water solution, make turbidity lower than the loading solution of 0.5 °, adopt macroporous resin D101 separator column, after wet method loading, standing, reach adsorption equilibrium to resin, take ethanol-water solution as elutriant, carry out gradient elution or etc. degree stepwise elution, collect the elutriant that oxidation resvertrol massfraction is greater than 40%, merge, concentrated, dry, obtain crude product, described standingly at room temperature carry out, the standing time is 0.5～1h;
(4) purification, to step (3) gained crude product repeating step (3), collects oxidation resvertrol massfraction and is more than or equal to 96.97% elutriant, merges, concentrated, dry, obtains described oxidation resvertrol.
2. method according to claim 1, is characterized in that: the described ramulus mori as raw material is dry Mulberry Twig.
3. method according to claim 1, is characterized in that: in step (2), described vacuum drying temperature is 25 ℃～30 ℃.
4. method according to claim 1, is characterized in that: in step (3), carry out gradient elution.
5. method according to claim 1, it is characterized in that: in step (3) and (4), stepwise elution such as degree of grade, 10% the concentration difference of take is a spacer, the elution volume of every section is 2～3 column volumes, eluent flow rate be 1～2 column volume/hour.
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| CN201210487502.2A CN102942456B (en) | 2012-11-26 | 2012-11-26 | Process for extracting oxyresveratrol from mulberry twigs |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060216253A1 (en) * | 2005-03-23 | 2006-09-28 | Time And Cross, Inc. | Whitening cosmetics containing morus alba extracts |
| CN101591680A (en) * | 2009-06-29 | 2009-12-02 | 西南大学 | The extracting method of oxidized resveratrol |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060216253A1 (en) * | 2005-03-23 | 2006-09-28 | Time And Cross, Inc. | Whitening cosmetics containing morus alba extracts |
| CN101591680A (en) * | 2009-06-29 | 2009-12-02 | 西南大学 | The extracting method of oxidized resveratrol |
Non-Patent Citations (2)
| Title |
|---|
| 张作法.桑枝活性成分分离纯化及其药理作用的研究.《中国优秀博士论文电子期刊网》.2011, |
| 桑枝活性成分分离纯化及其药理作用的研究;张作法;《中国优秀博士论文电子期刊网》;20110415;第100页第3段，第107页第2段，第115页最后一段至第116页第1段 * |
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