CN106632207B - A kind of preparation method of high-purity GCG - Google Patents
A kind of preparation method of high-purity GCG Download PDFInfo
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- CN106632207B CN106632207B CN201611198080.1A CN201611198080A CN106632207B CN 106632207 B CN106632207 B CN 106632207B CN 201611198080 A CN201611198080 A CN 201611198080A CN 106632207 B CN106632207 B CN 106632207B
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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
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
Abstract
The invention discloses the preparation methods of high-purity GCG a kind of, include the following steps: (1) using EGCG as raw material, and addition solvent is anti-to obtain reaction solution A；(2) by reaction solution A concentration to aqueous solution, ethyl acetate is added and is extracted, solution B is obtained；(3) solution B is taken, is added gradually in the chromatographic column equipped with macroporous absorbent resin, carries out gradient elution, choose the eluent containing target component, be concentrated into no alcohol taste, obtain concentrate C；(4) take concentrate C, n-butanol be added and is extracted, butanol solution is merged, is concentrated into medicinal extract, water is added and is dissolved to clearly, let cool crystallization, filtering drying to get high-purity GCG powder.The present invention is by after change of configuration occurs for the reaction conversion under hot conditions, by solvent extraction and macroporous adsorptive resin column technique, being prepared the GCG of high-purity using EGCG as raw material.
Description
Technical field
The present invention relates to field of medicine and chemical technology, and in particular to a kind of preparation method of high-purity GCG.
Background technique
Epigallo-catechin gallate (EGCG) (EGCG) is one of the major catechin in tealeaves, as natural anti-oxidation
Agent and free radical scavenger have apparent anti-aging, anti-mutation, cancer-resisting, prevention cardiovascular disease, radiation protection etc. a variety of
Biological activity is widely used in food processing, medicines and health protection and filed of daily-use chemical industry.However, due to its superpower oxidation and
Architectural characteristic, EGCG are also highly prone to the influence of environmental factor, such as easily aoxidize, are extremely easy in decomposition in alkali, in high temperature in air
Condition stability inferior difference, etc..
Nutgall catechin gallic acid ester (GCG) is a kind of isomer of EGCG, is deposited among natural green tea
It is considerably less measuring, it is difficult directly to directly obtain the very high GCG of purity from green tea, and then limit its application progress of research.
It can be appreciated that at present for the research of GCG or seldom, overwhelming majority research is concentrated on based on its isomers EGCG.
It is found in EGCG research process, is handled, EGCG can be effectively converted by some specific conditions
GCG, then by a series of purification techniques, the GCG sample of purity is high is finally obtained, and tentatively discovery is with shape existing for GCG configuration
Under formula, stability is good, is more advantageous to the exploitation and application of the kind.
Summary of the invention
Goal of the invention: in view of the problems of the existing technology, the present invention provides the preparation method of high-purity GCG a kind of, leads to
The available high-purity GCG of the preparation method is crossed, technique is simple and direct, and it is easy to operate, it is fully able to be satisfied with industrialization production requirements.
Technical solution: to achieve the goals above, a kind of preparation method of high-purity GCG as described in the present invention, including
Following steps:
(1) using EGCG as raw material, after solvent dissolution is added, it is transferred to thermostatic mixer, controls temperature in 60-100 DEG C
It obtains reaction solution A within reaction 4-6 days, lets cool spare；
(2) reaction solution A is let cool in 60-100 DEG C of concentration to aqueous solution, ethyl acetate is added and is extracted, merges
Aqueous continues concentration at 60-100 DEG C and removes remaining ethyl acetate, obtains solution B, let cool spare；
(3) solution B is taken, is added gradually in the chromatographic column equipped with macroporous absorbent resin, successively with water and different volumes point
Several ethyl alcohol carries out gradient elution, and 2-10 column volume of each gradient elution simultaneously receives each section eluent respectively, and selection contains
The eluent of target component is concentrated into no alcohol taste, obtains concentrate C, spare；
(4) concentrate C is taken, n-butanol is added and is extracted, merges butanol solution, is concentrated into medicinal extract at 60-100 DEG C,
Be added water be dissolved at 60-100 DEG C clearly, let cool it is complete to crystallization, filtering drying and crushing to get high-purity GCG powder.
Preferably, HPLC purity >=50% of step (1) described EGCG.
Preferably, step (1) solvent is selected from least one of purified water, methanol, ethyl alcohol or acetic acid, raw material with
The w/v of solvent is 1:(5-20) g/ml.
Preferably, step (2) the addition ethyl acetate carries out 3-5 extraction, the ethyl acetate being added every time with instead
Answering liquid A volume ratio is 0.2-2.
Preferably, step (2) solution B and raw material volume weight ratio are (5-20): 1ml/g.
Preferably, step (3) resin model be HPD-100, HPD-200, HPD-400, HPD-826 at least
It is a kind of.
Preferably, the ethyl alcohol of step (3) the different volumes score is 5-20% ethyl alcohol, 30-40% ethyl alcohol and 50-
90% ethyl alcohol.
Preferably, step (4) the addition n-butanol carries out 3-5 extraction, the n-butanol and concentrate C being added every time
Volume ratio is 0.2-2.
The envelope-bulk to weight ratio of step (4) water and medicinal extract is (0.5-10): 1ml/g.
The utility model has the advantages that compared with prior art, the present invention has the advantage that
1, the present invention is handled EGCG raw material by thermostatic mixer, and simple and safe operation, reaction temperature is moderate, no
Only avoid causes EGCG Partial digestion to be other non-targeted ingredients because temperature is excessively high, and also assures filling for configuration conversion
Divide property.
2, the present invention takes full advantage of the catechins such as EGCG, EC, ECG ingredient and GCG and dissolves in ethyl acetate and water
Property difference extractant ratio is adjusted by ethyl acetate abstraction technique, and control target component concentration, can be by major part
Catechin impurity component efficiently separate ethyl acetate layer, and GCG is retained in aqueous solution, thus reach efficiently separate it is pure
The purpose of change.
3, the present invention also applies macroreticular resin technology and purifies to target component, does not remove only most colors
Element further increases target component purity also by different gradients.
4, method according to the present invention can amplify in equivalent manner, and technique is simple and direct, easy to operate, be fully able to
It is satisfied with industrialization production requirements.
Specific embodiment
The invention will be further described with reference to embodiments.
Embodiment 1
(1) weighing the EGCG powder 50g that HPLC purity is 55% is raw material, after 10% methanol 1000ml dissolution is added, is turned
It moves on in 80 DEG C of thermostatic mixer water-baths, coreaction 5 days, obtains reaction solution A, let cool spare；
(2) ethyl acetate is added by reaction solution A in 70 DEG C of concentrations to no alcohol taste aqueous solution, after letting cool and carries out 3 extractions,
The ethyl acetate and reaction solution A volume ratio being added every time are 0.5, merge aqueous, continue concentration at 70 DEG C and remove remaining acetic acid
Ethyl ester, mending purified water makes final solution total volume 500ml, obtains solution B, lets cool spare；
(3) solution B is taken, (resin model HPD- in the chromatographic column equipped with 1000g macroporous absorbent resin is added gradually to
100 are mixed with HPD-400 by quality 1:1), gradient elution successively is carried out with water, 20% ethyl alcohol, 40% ethyl alcohol and 60% ethyl alcohol,
4 column volumes of each gradient elution simultaneously receive each section eluent respectively, choose the eluent containing target component, are concentrated into nothing
Alcohol taste obtains concentrate C, spare；
(4) concentrate C is taken, n-butanol is added and carries out 3 extractions, the n-butanol being added every time is with concentrate C volume ratio
0.2, merge butanol solution, is concentrated into medicinal extract at 80 DEG C, adds water to be dissolved at 80 DEG C clearly, (the envelope-bulk to weight ratio of water and medicinal extract
For 2ml/g), let cool complete to crystallization, filtering drying simultaneously crushes to get white powder 6.1g, is identified, is determined as GCG, passes through
HPLC detection, purity 92.3%.
Embodiment 2
(1) weighing the EGCG powder 50g that HPLC purity is 60% is raw material, after 10% ethyl alcohol 250ml dissolution is added, transfer
Into 60 DEG C of thermostatic mixer water-baths, coreaction 4 days, reaction solution A is obtained, is let cool spare；
(2) ethyl acetate is added by reaction solution A in 60 DEG C of concentrations to no alcohol taste aqueous solution, after letting cool and carries out 5 extractions
It takes, the ethyl acetate being added every time and reaction solution A volume ratio are 0.2, merge aqueous, continue concentration at 60 DEG C and remove remaining second
Acetoacetic ester, mending purified water makes final solution total volume 250ml, obtains solution B, lets cool spare；
(3) solution B is taken, (resin model HPD- in the chromatographic column equipped with 1000g macroporous absorbent resin is added gradually to
200) gradient elution successively, is carried out with water, 5% ethyl alcohol, 30% ethyl alcohol and 50% ethyl alcohol, 2 column volumes of each gradient elution are simultaneously
Each section eluent is received respectively, is chosen the eluent containing target component, is concentrated into no alcohol taste, obtains concentrate C, it is spare；
(4) concentrate C is taken, n-butanol is added and carries out 5 extractions, the n-butanol being added every time is with concentrate C volume ratio
0.5, merge butanol solution, is concentrated into medicinal extract at 60 DEG C, adds water to be dissolved at 60 DEG C clearly, (the envelope-bulk to weight ratio of water and medicinal extract
For 10ml/g), let cool complete to crystallization, filtering drying simultaneously crushes to get white powder 5.4g, is identified, is determined as GCG, passes through
HPLC detection, purity 94.5%.
Embodiment 3
(1) weighing the EGCG powder 50g that HPLC purity is 58% is raw material, after 10% acetic acid 500ml dissolution is added, transfer
Into 100 DEG C of thermostatic mixer water-baths, coreaction 6 days, reaction solution A is obtained, is let cool spare；
(2) ethyl acetate is added by reaction solution A in 100 DEG C of concentrations to aqueous solution, after letting cool and carries out 4 extractions, every time
The ethyl acetate and reaction solution A volume ratio of addition are 2, merge aqueous, continue concentration at 100 DEG C and remove remaining ethyl acetate,
Mending purified water makes final solution total volume 1000ml, obtains solution B, lets cool spare；
(3) solution B is taken, (resin model HPD- in the chromatographic column equipped with 1000g macroporous absorbent resin is added gradually to
826) gradient elution successively, is carried out with water, 20% ethyl alcohol, 40% ethyl alcohol and 90% ethyl alcohol, 2 column volumes of each gradient elution are simultaneously
Each section eluent is received respectively, is chosen the eluent containing target component, is concentrated into no alcohol taste, obtains concentrate C, it is spare；
(4) concentrate C is taken, n-butanol is added and carries out 4 extractions, the n-butanol being added every time is with concentrate C volume ratio
2, merge butanol solution, is concentrated into medicinal extract at 100 DEG C, adds water to be dissolved at 100 DEG C clearly, (the envelope-bulk to weight ratio of water and medicinal extract
For 0.5ml/g), let cool complete to crystallization, filtering drying simultaneously crushes to get white powder 7.2g, is identified, is determined as GCG, passes through
HPLC detection, purity 90.3%.
Claims (1)
1. a kind of preparation method of high-purity GCG, which comprises the steps of:
(1) using EGCG as raw material, after solvent dissolution is added, it is transferred to thermostatic mixer, control temperature is reacted in 60-100 DEG C
It obtains reaction solution A within 4-6 days, lets cool spare；
(2) reaction solution A is let cool in 60-100 DEG C of concentration to aqueous solution, ethyl acetate is added and is extracted, water is merged
Liquid continues concentration at 60-100 DEG C and removes remaining ethyl acetate, obtains solution B, let cool spare；
(3) solution B is taken, is added gradually in the chromatographic column equipped with macroporous absorbent resin, successively with water and different volumes score
Ethyl alcohol carries out gradient elution, and 2-10 column volume of each gradient elution simultaneously receives each section eluent respectively, and selection contains target
The eluent of ingredient is concentrated into no alcohol taste, obtains concentrate C, spare；
(4) concentrate C is taken, n-butanol is added and is extracted, merges butanol solution, medicinal extract is concentrated at 60-100 DEG C, is added
Water is dissolved to clearly at 60-100 DEG C, let cool it is complete to crystallization, filtering drying and crush to get high-purity GCG powder；
HPLC purity >=50% of step (1) described EGCG；Step (1) solvent is selected from purified water, with methanol, ethyl alcohol or vinegar
The w/v of the mixed solution of any one in three kinds of acid, raw material and solvent is 1:(5-20) g/ml；Step (2) is described to be added
Enter ethyl acetate and carry out 3-5 extraction, the ethyl acetate being added every time and reaction solution A volume ratio are 0.2-2；Step (2) is described
Solution B and raw material volume weight ratio are (5-20): 1 ml/g；Step (3) resin model be HPD-100, HPD-200,
At least one of HPD-400, HPD-826；The ethyl alcohol of step (3) the different volumes score is 5-20% ethyl alcohol, 30-40% second
Pure and mild 50-90% ethyl alcohol；Step (4) the addition n-butanol carries out 3-5 extraction, the n-butanol and concentrate C body being added every time
Product is than being 0.2-2；The envelope-bulk to weight ratio of step (4) water and medicinal extract is (0.5-10): 1 ml/g.
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| CN201611198080.1A CN106632207B (en) | 2016-12-22 | 2016-12-22 | A kind of preparation method of high-purity GCG |
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| CN201611198080.1A CN106632207B (en) | 2016-12-22 | 2016-12-22 | A kind of preparation method of high-purity GCG |
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| CN106632207B true CN106632207B (en) | 2019-04-16 |
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| CN112979604A (en) * | 2019-12-18 | 2021-06-18 | 江苏天晟药业股份有限公司 | Preparation method of gallocatechin gallate |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101053358A (en) * | 2006-04-14 | 2007-10-17 | 株式会社伊藤园 | Bottled green tea beverage |
| CN102120738A (en) * | 2010-01-11 | 2011-07-13 | 温尧林 | Preparation method of nonepicatechin monomer |
| CN105601606A (en) * | 2016-03-08 | 2016-05-25 | 浙江省计量科学研究院 | Method for preparing high-purity gallnut catechin gallate (GCG) |
-
2016
- 2016-12-22 CN CN201611198080.1A patent/CN106632207B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101053358A (en) * | 2006-04-14 | 2007-10-17 | 株式会社伊藤园 | Bottled green tea beverage |
| CN102120738A (en) * | 2010-01-11 | 2011-07-13 | 温尧林 | Preparation method of nonepicatechin monomer |
| CN105601606A (en) * | 2016-03-08 | 2016-05-25 | 浙江省计量科学研究院 | Method for preparing high-purity gallnut catechin gallate (GCG) |
Non-Patent Citations (3)
| Title |
|---|
| 分离制备儿茶素及EGCG的研究进展;李慧星 等，;《河北农业科学》;20081231;第12卷(第6期);第92页 左栏倒数第1段 |
| 大孔吸附树脂对酯型儿茶素吸附性能的研究;钟世安 等，;《离子交换与吸附》;20070831;第23卷(第5期);第392页 倒数第2段，摘要 |
| 高速逆流色谱分离同分异构体;陈理 等，;《色谱》;20061130;第24卷(第6期);第570-573页 |
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