CN107176941A - A kind of method that epicatechin is extracted from Testa sojae atricolor - Google Patents
A kind of method that epicatechin is extracted from Testa sojae atricolor Download PDFInfo
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- CN107176941A CN107176941A CN201710589919.2A CN201710589919A CN107176941A CN 107176941 A CN107176941 A CN 107176941A CN 201710589919 A CN201710589919 A CN 201710589919A CN 107176941 A CN107176941 A CN 107176941A
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- eluent
- sojae atricolor
- epicatechin
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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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- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a kind of method that epicatechin is extracted from Testa sojae atricolor, it is to be filtered after Testa sojae atricolor is extracted, filtrate decompression is concentrated, concentrated aqueous solution is obtained.By Sepabeads SP700 resin columns on concentrated aqueous solution, eluted successively with the ethanol solution of pure water and 60%, collected volume concentration is 60% ethanol solution.By the posts of Sephadex LH 20 on concentrated aqueous solution, eluted successively with volumetric concentration for 25%, 50%, 60%, 70%, 80%, 90% methanol solution, collected volume concentration is used as eluent I for 25% whole eluents, collected volume concentration having at 280nm corresponding to 50% methanol solution absorbs and the eluent without absorption is used as eluent II at 360nm, merge eluent I and eluent II is concentrated under reduced pressure, the resulting epicatechin aqueous solution is dried, and obtains Testa sojae atricolor epicatechin.Detect Testa sojae atricolor epicatechin purity up to 95% with efficient liquid phase.
Description
Technical field
The present invention relates to effective ingredients in plant extraction, a kind of preparation high-purity epicatechin from Testa sojae atricolor is particularly belonged to
Method.
Background technology
Epicatechin (- epicatechin) is OPC (proanthocyanidin) one kind, is 3- hydroxyl flavane
01 derivatives, the polyatomic phenol material with C6-C3-C6 structures, its basic structure is α-phenyl benzopyrane, i.e., in two benzene
It is connected between ring with three carbochains, one of carbon atom connects into pyranoid ring with the oxygen atom on A rings.Now it has proven convenient that youngster
Theine class material has a variety of effects beneficial to health, including removes free radical, anti-oxidant, anticancer, anti-inflammatory, antibacterial, anti-purple
Outside line, prevention of cardiovascular disease etc..Therefore, catechin is considered as the natural production of the ideal of development function food and plant sex medicine
Thing.[Peng,Han,etal.Journal of Functional Foods(2017),32,296-312.]
Black soya bean is also known as horsebean, winter bean, self-sown beans, and most of black soya bean seeds are smaller, are commonly called as Semen glycines sojae.How flat the seed of black soya bean is
Oval, the oblate, kidney shape, oblong, round shaped grain are less.The black soya bean germ plasm resource of China is extremely abundant, according to statistics, and the whole nation is collected altogether
More than 22000 parts of soybean is preserved, wherein black soya bean cultivar just there are more than 2800 kinds, accounts for the 13.2% of whole Soybean Germplasms, extensively
It is general to be distributed in 27 provinces, a cities, it is especially more with Shanxi, Hebei, Shaanxi cultivation, wherein with Beijing early soybean, Huanghuai Summer Sowing Soybean in China most
To be common.So far, the specific chemical composition report of Testa sojae atricolor, in terms of being concentrated mainly on anthocyanidin, and for OPC
Research it is less.[Jin Hwan Lee,etal.Food Chemistry.2009,112(1),226-231.].For with black soya bean
Skin is not reported for the method that raw material prepares epicatechin.
The content of the invention
The invention aims to provide a kind of method of quick, the high efficiency extraction epicatechin from Testa sojae atricolor.
In order to solve the above-mentioned technical problem, the present invention provides a kind of method that epicatechin is extracted from Testa sojae atricolor, including
Following steps：
Testa sojae atricolor is taken, is crushed, 40-60 mesh sieves are crossed；It is put into 50-70% aqueous acetone solutions, adjusts in 1g/10-50mL ratios
PH2.0 is saved, 3-5h is extracted, extract solution is filtered with filter paper, filtrate is obtained；Filtrate decompression is concentrated, crude extract is obtained；Crude extract wet method
Upper prop, applied sample amount 40-60mg crude extracts/g resins；Be eluted to the outflow of no turbid solution with distilled water first, Liu Su≤4mL/min and
PH stable is in 6-8, then is eluted with the ethanol water of eluant, eluent 60% colourless and pH is in 5-7 to efflux, and eluent rotation is steamed
Hair, is freeze-dried slightly to get sample；Sephadex LH-20 posts in the thick wet method that gets sample, successively with 25%, 50%, 60%, 70%,
80%th, 90% methanol aqueous solution is eluted, and consumption is three times column volume, and flow velocity is 50d/min；Collect 25% methanol-water
Solution eluent as eluent I, collect 50% methanol aqueous solution latter half eluent (have absorption at 280nm and
Eluent part without absorption at 360nm) it is used as eluent II；Merge eluent I and eluent II is concentrated, reclaim first
Alcohol, obtains Testa sojae atricolor epicatechin.
The ratio of the Testa sojae atricolor and aqueous acetone solution is preferably 1g/10-40mL.
Described pH adjusting agent is preferably acetic acid.
Described extraction time is preferably 3h.
The method that the present invention extracts epicatechin from Testa sojae atricolor, has the following advantages that：
1. the present invention is by raw material of Testa sojae atricolor, separation prepares former catechin, can not only increase the added value of black soya bean industry,
Economic benefit is brought, resource can also be made to be fully used.
2. the present invention uses Sephadex LH-20 to prepare epicatechin, efficiency high for column packing separation, and can keep it
Original structure and activity, resulting epicatechin purity are high, and solvent is repeatable to be utilized, it is to avoid pollution and waste.
Embodiment
Embodiment 1 extracts epicatechin from Testa sojae atricolor：
By the blue or green benevolence Testa sojae atricolor of the black skin in 10g Shanxi Province, crush, cross 60 mesh sieves.It is put into 1g/10mL ratios in 70% acetone,
With vinegar acid for adjusting pH 2.0,3h is extracted, extract solution is filtered with filter paper, filter residue is repeated into aforesaid operations, extract 2 times (i.e. with filter
Slag represents Testa sojae atricolor and operated as above), merge 3 gained filtrates, filtrate decompression is concentrated, crude extract is obtained.Applied sample amount 50mg
Crude extract/g resins, column type 2.5cm × 50cm, wet method upper prop.Eluted and flowed out up to without turbid solution with distilled water first, Liu Su≤
4mL/min and pH stable is 7 or so, then with the ethanol elution of eluant, eluent 60% to the colourless pH of efflux in 6 or so, eluent rotation
Turn evaporation, be freeze-dried slightly to get sample.From volume be 1.5cm × 30cm Sephadex LH-20 cylinders, wet method upper prop, according to
Secondary to use 25%, 50%, 60%, 70%, 80%, 90% methanol solution is eluted, and consumption respectively (divides for three times column volume
Wei 180ml)；Flow velocity is 50d/min, and it is eluent I to collect the whole eluents of 25% methanol, collects 50% methanol aqueous solution
Latter half eluent (have at 280nm absorption and at 360nm the eluent part without absorption) is as eluent II.Merge
Eluent I and eluent II are concentrated, and are reclaimed methanol, are obtained Testa sojae atricolor epicatechin 0.39g.Above-mentioned Testa sojae atricolor epicatechin warp
High performance liquid chromatography detects that purity is ＞ 95%.
Embodiment 2 extracts epicatechin from Testa sojae atricolor：
By the blue or green benevolence Testa sojae atricolor of 50g Shandong Province pearlite layer, selected impurity elimination crushed 40 mesh sieves after processing.Put in 1g/40mL ratios
Enter in 70% acetone, with vinegar acid for adjusting pH 2.0, extract 3h, extract solution is filtered with filter paper, filter residue is repeated into aforesaid operations, soak
Carry 2 times (representing Testa sojae atricolor with filter residue to be operated as above), merge 3 gained filtrates, filtrate decompression is concentrated, obtains and slightly carries
Thing.Applied sample amount 50mg crude extracts/g resins, column type 4cm × 50cm, wet method upper prop.Eluted first with distilled water until without turbid solution
Outflow, Liu Su≤4mL/min and pH stable is 7 or so, then it is left 6 with the ethanol elution of eluant, eluent 60% to the colourless pH of efflux
The right side, eluent rotary evaporation is freeze-dried slightly to get sample.It is wet from the Sephadex LH-20 cylinders that volume is 3cm × 30cm
Method upper prop, is eluted with 25%, 50%, 60%, 70%, 80%, 90% methanol solution successively, and consumption is respectively three times post
Volume (being respectively 360ml)；Flow velocity is 60d/min, and it is eluent I to collect the whole eluents of 25% methanol, collects 50%
Methanol aqueous solution latter half eluent (have at 280nm absorption and at 360nm the eluent part without absorption) is as washing
De- liquid II.Merge eluent I and eluent II is concentrated, reclaim methanol, obtain Testa sojae atricolor epicatechin 1.26g.Above-mentioned black soya bean
Epidermis catechin detects that purity is ＞ 95% through high performance liquid chromatography.
Embodiment 3 extracts epicatechin from Testa sojae atricolor：
By the blue or green benevolence Testa sojae atricolor of the black skin in 10g Shanxi Province, selected impurity elimination crushed 60 mesh sieves after processing.Put in 1g/10mL ratios
Enter in 50% acetone, with vinegar acid for adjusting pH 2.0, extract 3h, extract solution is filtered with filter paper, filter residue is repeated into aforesaid operations, soak
Carry 2 times (representing Testa sojae atricolor with filter residue to be operated as above), merge 3 gained filtrates, filtrate decompression is concentrated, obtains and slightly carries
Thing.Applied sample amount 50mg crude extracts/g resins, column type 2.5cm × 50cm, wet method upper prop.Eluted first with distilled water until without muddiness
Liquid flows out, Liu Su≤4mL/min and pH stable is 7 or so, then with the ethanol elution of eluant, eluent 60% to the colourless pH of efflux 6
Left and right, eluent rotary evaporation is freeze-dried slightly to get sample.From the Sephadex LH-20 posts that volume is 1.5cm × 30cm
Body, wet method upper prop is eluted, consumption is respectively with 25%, 50%, 60%, 70%, 80%, 90% methanol solution successively
Three times column volume (being respectively 180ml)；Flow velocity is 50d/min, and it is eluent I to collect the whole eluents of 25% methanol, is received
Collect 50% methanol aqueous solution latter half eluent (have at 280nm absorption and at 360nm the eluent part without absorption)
It is used as eluent II.Merge eluent I and eluent II is concentrated, reclaim methanol, obtain Testa sojae atricolor epicatechin 0.48g.On
State Testa sojae atricolor epicatechin to detect through high performance liquid chromatography, purity is ＞ 92%.
Embodiment 4 extracts epicatechin from Testa sojae atricolor：
By the blue or green benevolence Testa sojae atricolor of 50g Shandong Province pearlite layer, selected impurity elimination crushed 40 mesh sieves after processing.Put in 1g/40mL ratios
Enter in 70% acetone, pH2.0 is adjusted with acetic acid, extract 3h, extract solution is filtered with filter paper, filter residue is repeated into aforesaid operations, soak
Carry 2 times (representing Testa sojae atricolor with filter residue to be operated as above), merge 3 gained filtrates, filtrate decompression is concentrated, obtains and slightly carries
Thing.Applied sample amount 50mg crude extracts/g resins, column type 4cm × 50cm, wet method upper prop.Eluted first with distilled water until without turbid solution
Outflow, Liu Su≤4mL/min and pH stable is 7 or so, then it is left 6 with the ethanol elution of eluant, eluent 50% to the colourless pH of efflux
The right side, eluent rotary evaporation is freeze-dried slightly to get sample.It is wet from the Sephadex LH-20 cylinders that volume is 3cm × 30cm
Method upper prop, is eluted with 25%, 50%, 60%, 70%, 80%, 90% methanol solution successively, and consumption is respectively three times post
Volume (being respectively 360ml)；Flow velocity is 60d/min, and it is eluent I to collect the whole eluents of 25% methanol, collects 50%
Methanol aqueous solution latter half eluent (have at 280nm absorption and at 360nm the eluent part without absorption) is as washing
De- liquid II.Merge eluent I and eluent II is concentrated, reclaim methanol, obtain Testa sojae atricolor epicatechin 1.88g.Above-mentioned black soya bean
Epidermis catechin detects that purity is ＞ 90% through high performance liquid chromatography.
It is described above, only it is the preferable embodiment in part of the present invention, any formal limit not is done to the present invention
System.
Claims (4)
1. a kind of method that epicatechin is extracted from Testa sojae atricolor, it is characterised in that comprise the following steps：Testa sojae atricolor is taken, is crushed,
Cross 40-60 mesh sieves；It is put into 1g/10-50mL ratios in 50-70% aqueous acetone solutions, adjusts pH2.0, is extracted 3-5h, will extract
Liquid is filtered with filter paper, obtains filtrate；Filtrate decompression is concentrated, crude extract is obtained；Crude extract wet method upper prop, applied sample amount 40-60mg is slightly carried
Thing/g resins；First it is eluted to the outflow of no turbid solution with distilled water, Liu Su≤4mL/min and pH stable is in 6-8, then use eluant, eluent
60% ethanol water elutes colourless and pH is in 5-7 to efflux, and eluent rotary evaporation is freeze-dried slightly to get sample；Slightly carry
Sephadex LH-20 posts in sample wet method, are washed with 25%, 50%, 60%, 70%, 80%, 90% methanol aqueous solution successively
De-, consumption is three times column volume, and flow velocity is 50d/min；25% methanol aqueous solution eluent is collected as eluent I, is received
Collect 50% methanol aqueous solution latter half eluent as eluent II；Merge eluent I and eluent II is concentrated, reclaim
Methanol, obtains Testa sojae atricolor epicatechin.
2. a kind of method that epicatechin is extracted from Testa sojae atricolor as claimed in claim 1, it is characterised in that the Testa sojae atricolor
Ratio with aqueous acetone solution is 1g/10-40mL.
3. according to the method that epicatechin is extracted in a kind of Testa sojae atricolor described in claim 1, it is characterised in that described pH is adjusted
Section agent is acetic acid.
4. according to the method that epicatechin is extracted in a kind of Testa sojae atricolor described in claim 1, it is characterised in that described extraction
Time is 3h.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710589919.2A CN107176941A (en) | 2017-07-19 | 2017-07-19 | A kind of method that epicatechin is extracted from Testa sojae atricolor |
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| CN201710589919.2A CN107176941A (en) | 2017-07-19 | 2017-07-19 | A kind of method that epicatechin is extracted from Testa sojae atricolor |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20220043660A (en) | 2020-09-29 | 2022-04-05 | (주)아모레퍼시픽 | New soybean variety comprising high content of epicatechin |
-
2017
- 2017-07-19 CN CN201710589919.2A patent/CN107176941A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 刘占云等: "HPLC法测定不同产地黑豆皮中表儿茶素的含量", 《食品研究与开发》 * |
| 张翠等: "黑豆种皮的酚酸类成分研究", 《中草药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20220043660A (en) | 2020-09-29 | 2022-04-05 | (주)아모레퍼시픽 | New soybean variety comprising high content of epicatechin |
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Application publication date: 20170919 |