CN104016958B - A kind of preparation method of high-purity epigallocatechin-3-gallate - Google Patents
A kind of preparation method of high-purity epigallocatechin-3-gallate Download PDFInfo
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- CN104016958B CN104016958B CN201410205827.6A CN201410205827A CN104016958B CN 104016958 B CN104016958 B CN 104016958B CN 201410205827 A CN201410205827 A CN 201410205827A CN 104016958 B CN104016958 B CN 104016958B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
Abstract
The present invention relates to a kind of preparation method of easy and simple to handle, high-purity epigallocatechin-3-gallate that production cost is low.The method is made up of the following step: get tea-polyphenol, is dissolved in water, and filters, get filtrate, add mixing solutions abstraction impurity removal containing ethyl acetate isopyknic with it, water intaking layer liquid, concentrated, join on nonpolarity macroporous adsorptive resins chromatographic column and adsorb, ethanolic soln wash-out, collect elutriant, reclaim ethanol and concentrate, joining on polar macroporous adsorbent resin column and adsorb, ethanolic soln wash-out, collects elutriant, reclaims ethanol and concentrates, lyophilize, to obtain final product.Adopt preparation table nutgall catechin gallic acid ester of the present invention, product purity is high, production efficiency is high.
Description
Technical field
The present invention relates to tea technology field, especially tea leaf extract technical field.
Background technology
NVP-XAA 723 (hereinafter referred to as EGCG) is a kind of composition extracted from Chinese green tea, it is the main activity of green tea and water-soluble components, it is the component that in catechin, content is the highest, account for the 9%-13% of green tea gross weight, because have special stereochemical structure, EGCG has very strong anti-oxidant activity, anti-oxidant activity is at least ascorbic more than 100 times, it is 25 times of vitamin-E, can Cell protection and DNA undermined, this infringement is believed and cancer, heart disease is relevant with other major diseases, these effects of EGCG ascribe their removing (anti-oxidant) ability to oxyradical to.
EGCG act as important role in anticancer and cardiovascular disorder.In addition, it is also used as the reversal agent of multi-drug resistance of the tumor, can improve cancer cells to the susceptibility of chemotherapy and the toxicity alleviated heart.
EGCG can make antioxygen, antibacterial, fresh-keeping, deodorant in foodstuffs industry; Daily chemical products is done the preservative of specific function, skin-protecting agent.
The physico-chemical property of EGCG is as follows:
CASNO.：989-51-5
Molecular formula: C22H18O11
Molecular weight: 458.38
Structural formula:
EGCG is one of main component in commodity tea-polyphenol, and tea-polyphenol is with tealeaves (CamelliaSinensis.L) for raw material, through the polyphenolic compound based on catechin extracted.
In prior art, the extraction purification of EGCG mainly adopts the methods such as organic solvent extraction and silicagel column separation, and cost is high, seriously polluted, is unfavorable for large production operation.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of preparation method of high-purity epigallocatechin-3-gallate, and the method is easy and simple to handle, environmental friendliness, production efficiency are high, is applicable to the large manufacture of industry.
In order to solve the problems of the technologies described above, the present invention proposes following technical proposal:
Get tea-polyphenol, be dissolved in water, filter, get filtrate, add mixing solutions abstraction impurity removal containing ethyl acetate isopyknic with it, water intaking layer liquid, concentrated, join on nonpolarity macroporous adsorptive resins chromatographic column and adsorb, be that 5%-30% ethanolic soln carries out wash-out by volume percent, collect the elutriant of 3-8 times amount column volume, reclaim ethanol and concentrated, join on polar macroporous adsorbent resin column and adsorb, be that 10%-50% ethanolic soln carries out wash-out by volume percent, collect the elutriant of 3-8 times amount column volume, reclaim ethanol and concentrated, lyophilize, to obtain final product.
Described tea-polyphenol is take tealeaves as raw material, through the polyphenolic compound based on catechin extracted, wherein containing NVP-XAA 723.
Tea-polyphenol can adopt following method to prepare: get green tea, and the ratio of 1:7-10 adds water and carries out temperature leaching in mass ratio, and soaking temperature 90 DEG C, soaks 3 times, each 40 minutes, filtration, merging filtrate, concentrate drying and get final product.
The described mixing solutions containing ethyl acetate is selected from ethyl acetate-chloroform mixing solutions that volume ratio is 1:2-5, volume ratio is the ethyl acetate-dichloromethane of 1:2-5 or one in the ethyl acetate-light petrol mixing solutions of volume ratio 1:2-5.
Described nonpolar macroporous adsorption resin is selected from one or more in XAD-1, XAD-4, HP-20, D4006, D101 type macroporous adsorbent resin.
Described polar macroporous adsorption resin is selected from one or more in XAD-9, XAD-10, NAK-9, DA201, AB-8 type macroporous adsorbent resin.
In described nonpolar macroporous adsorption resin and load solution, the mass ratio of contained solid is 10-40:1.
In described polar macroporous adsorption resin and load solution, the mass ratio of contained solid is 10-40:1.
The pH value of described nonpolar macroporous adsorption resin wash-out ethanolic soln is 3-7.
The pH value of described polar macroporous adsorption resin wash-out ethanolic soln is 3-7.
The present invention is with common tea polyphenol for raw material, and utilize series connection macroporous resin adsorption column chromatography technology, using aqueous ethanolic solution as eluting solvent, the EGCG in tea-polyphenol is enriched to more than 98%, yield is high, speed is fast, can be used for factory and prepares EGCG on a large scale.The invention solves the defect that EGCG production efficiency is low, cost is high in the past.
Macroporous resin during this law is bright can be reused and regenerate, and the widespread use for EGCG provides a kind of more practical preparation method.
In order to better set forth technical scheme of the present invention, below in conjunction with embodiment, the present invention is further illustrated, but protection domain of the presently claimed invention is not limited to the following example.
Embodiment
Embodiment 1
Get commercially available tea-polyphenol (EGCG content is denoted as 30%) 100Kg, be dissolved in water, filter, get filtrate, add ethyl acetate-chloroform mixing solutions isopyknic with it (ethyl acetate: chloroform=1:2) abstraction impurity removal, water intaking layer liquid, concentrated, join on XAD-1 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 10:1), be that 5% ethanolic soln (pH value is 3) carries out wash-out by volume percent, collect the elutriant of 3 times amount column volumes, reclaim ethanol and concentrate, join on XAD-9 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 10:1), be that 10% ethanolic soln (pH value is 3) carries out wash-out by volume percent, collect the elutriant of 3 times amount column volumes, reclaim ethanol and concentrate, lyophilize, obtain EGCG25.6Kg.
With reference to method one " detection-HPLC method of Catechin in Tea class " in GB/T8313-2008 standard, adopt external standard direct quantitative, the purity recording EGCG is 98.3%.
Embodiment 2
Get commercially available tea-polyphenol (EGCG content is denoted as 40%) 100Kg, be dissolved in water, filter, get filtrate, add ethyl acetate-chloroform mixing solutions isopyknic with it (ethyl acetate: chloroform=1:5) abstraction impurity removal, water intaking layer liquid, concentrated, join on XAD-4 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 40:1), be that 30% ethanolic soln (pH value is 7) carries out wash-out by volume percent, collect the elutriant of 8 times amount column volumes, reclaim ethanol and concentrate, join on XAD-10 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 40:1), be that 50% ethanolic soln (pH value is 7) carries out wash-out by volume percent, collect the elutriant of 8 times amount column volumes, reclaim ethanol and concentrate, lyophilize, obtain EGCG36.2Kg.
With reference to method one " detection-HPLC method of Catechin in Tea class " in GB/T8313-2008 standard, adopt external standard direct quantitative, the purity recording EGCG is 98.2%.
Embodiment 3
Get commercially available tea-polyphenol (EGCG content is denoted as 30%) 100Kg, be dissolved in water, filter, get filtrate, add ethyl acetate-chloroform mixing solutions isopyknic with it (ethyl acetate: chloroform=1:3) abstraction impurity removal, water intaking layer liquid, concentrated, join on HP-20 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 20:1), be that 20% ethanolic soln (pH value is 5) carries out wash-out by volume percent, collect the elutriant of 5 times amount column volumes, reclaim ethanol and concentrate, join on NAK-9 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 20:1), be that 40% ethanolic soln (pH value is 5) carries out wash-out by volume percent, collect the elutriant of 5 times amount column volumes, reclaim ethanol and concentrate, lyophilize, obtain EGCG26.7Kg.
With reference to method one " detection-HPLC method of Catechin in Tea class " in GB/T8313-2008 standard, adopt external standard direct quantitative, the purity recording EGCG is 98.1%.
Embodiment 4
Get tea-polyphenol (preparing by embodiment 6) 100Kg, be dissolved in water, filter, get filtrate, add ethyl acetate-dichloromethane mixing solutions isopyknic with it (ethyl acetate: methylene dichloride=1:2) abstraction impurity removal, water intaking layer liquid, concentrated, join on D4006 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 20:1), be that 20% ethanolic soln (pH value is 4) carries out wash-out by volume percent, collect the elutriant of 5 times amount column volumes, reclaim ethanol and concentrate, join on DA201 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 30:1), be that 40% ethanolic soln (pH value is 3) carries out wash-out by volume percent, collect the elutriant of 5 times amount column volumes, reclaim ethanol and concentrate, lyophilize, obtain EGCG17.8Kg.
With reference to method one " detection-HPLC method of Catechin in Tea class " in GB/T8313-2008 standard, adopt external standard direct quantitative, the purity recording EGCG is 98.7%.
Embodiment 5
Get tea-polyphenol (preparing by embodiment 8) 100Kg, be dissolved in water, filter, get filtrate, add ethyl acetate-light petrol mixing solutions isopyknic with it (ethyl acetate: sherwood oil=1:2) abstraction impurity removal, water intaking layer liquid, concentrated, join on D101 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 20:1), be that 10% ethanolic soln (pH value is 4) carries out wash-out by volume percent, collect the elutriant of 5 times amount column volumes, reclaim ethanol and concentrate, join on AB-8 type macroporous adsorptive resins and adsorb (contained by macroporous adsorbent resin and load solution, the mass ratio of solid is 30:1), be that 50% ethanolic soln (pH value is 3) carries out wash-out by volume percent, collect the elutriant of 5 times amount column volumes, reclaim ethanol and concentrate, lyophilize, obtain EGCG16.4Kg.
With reference to method one " detection-HPLC method of Catechin in Tea class " in GB/T8313-2008 standard, adopt external standard direct quantitative, the purity recording EGCG is 99.5%.
Embodiment 6
The preparation of tea-polyphenol
Get green tea 10Kg, in mass ratio the ratio of 1:7 add water carry out temperature leaching, soaking temperature 90 DEG C, soaks 3 times, each 40 minutes, filtration, merging filtrate, namely concentrate drying obtains tea-polyphenol 2.4Kg.
Embodiment 7
The preparation of tea-polyphenol
Get green tea 10Kg, in mass ratio the ratio of 1:10 add water carry out temperature leaching, soaking temperature 90 DEG C, soaks 3 times, each 40 minutes, filtration, merging filtrate, namely concentrate drying obtains tea-polyphenol 2.5Kg.
Embodiment 8
The preparation of tea-polyphenol
Get green tea 10Kg, in mass ratio the ratio of 1:8 add water carry out temperature leaching, soaking temperature 90 DEG C, soaks 3 times, each 40 minutes, filtration, merging filtrate, namely concentrate drying obtains tea-polyphenol 2.4Kg.
Claims (6)
1. a preparation method for high-purity epigallocatechin-3-gallate, is characterized in that described method is made up of the following step:
Get tea-polyphenol, be dissolved in water, filter, get filtrate, add mixing solutions abstraction impurity removal containing ethyl acetate isopyknic with it, water intaking layer liquid, concentrated, join on nonpolarity macroporous adsorptive resins chromatographic column and adsorb, be that 5%-30% ethanolic soln carries out wash-out by volume percent, collect the elutriant of 3-8 times amount column volume, reclaim ethanol and concentrated, join on polar macroporous adsorbent resin column and adsorb, be that 10%-50% ethanolic soln carries out wash-out by volume percent, collect the elutriant of 3-8 times amount column volume, reclaim ethanol and concentrated, lyophilize, to obtain final product;
The described mixing solutions containing ethyl acetate is selected from ethyl acetate-chloroform mixing solutions that volume ratio is 1:2-5, volume ratio is the ethyl acetate-dichloromethane of 1:2-5 or one in the ethyl acetate-light petrol mixing solutions of volume ratio 1:2-5;
The pH value of described nonpolar macroporous adsorption resin wash-out ethanolic soln is 3-7;
The pH value of described polar macroporous adsorption resin wash-out ethanolic soln is 3-7.
2. the preparation method of a kind of high-purity epigallocatechin-3-gallate according to claim 1, it is characterized in that, described tea-polyphenol is take tealeaves as raw material, through the polyphenolic compound based on catechin extracted, wherein containing NVP-XAA 723.
3. the preparation method of a kind of high-purity epigallocatechin-3-gallate according to claim 1, it is characterized in that, described nonpolar macroporous adsorption resin is selected from one or more in XAD-1, XAD-4, HP-20, D4006, D101 type macroporous adsorbent resin.
4. the preparation method of a kind of high-purity epigallocatechin-3-gallate according to claim 1, it is characterized in that, described polar macroporous adsorption resin is selected from one or more in XAD-9, XAD-10, NAK-9, DA201, AB-8 type macroporous adsorbent resin.
5. the preparation method of a kind of high-purity epigallocatechin-3-gallate according to claim 1, is characterized in that, in described nonpolar macroporous adsorption resin and load solution, the mass ratio of contained solid is 10-40:1.
6. the preparation method of a kind of high-purity epigallocatechin-3-gallate according to claim 1, is characterized in that, in described polar macroporous adsorption resin and load solution, the mass ratio of contained solid is 10-40:1.
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Citations (4)
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| CN1286252A (en) * | 1999-08-16 | 2001-03-07 | 弗·哈夫曼-拉罗切有限公司 | Process for preparing epigallocatechin gallate |
| CN1421426A (en) * | 2002-10-30 | 2003-06-04 | 湖北省化学研究院 | New process of preparing tea polyphenol with high catechin content and low caffine content |
| CN101074224A (en) * | 2007-04-13 | 2007-11-21 | 桂林莱茵生物科技股份有限公司 | Production of high-content EGCG |
| CN101643466A (en) * | 2009-06-02 | 2010-02-10 | 江苏天晟药业有限公司 | Epigallo-catechin gallate (EGCG) with high purity and preparation method thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1286252A (en) * | 1999-08-16 | 2001-03-07 | 弗·哈夫曼-拉罗切有限公司 | Process for preparing epigallocatechin gallate |
| CN1421426A (en) * | 2002-10-30 | 2003-06-04 | 湖北省化学研究院 | New process of preparing tea polyphenol with high catechin content and low caffine content |
| CN101074224A (en) * | 2007-04-13 | 2007-11-21 | 桂林莱茵生物科技股份有限公司 | Production of high-content EGCG |
| CN101643466A (en) * | 2009-06-02 | 2010-02-10 | 江苏天晟药业有限公司 | Epigallo-catechin gallate (EGCG) with high purity and preparation method thereof |
Non-Patent Citations (2)
| Title |
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| 儿茶素富集和EGCG单体纯化新工艺研究;张星海等;《茶叶》;20021231;第28卷(第3期);第136-137页 * |
| 大孔吸附树脂法分离纯化表没食子儿茶素没食子酸酯(EGCG)的研究;庄俊钰等;《现代食品科技》;20101231;第26卷(第11期);第1204-1206、1249页 * |
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