CN105586378A - Preparation method for assisting in extracting stibene glucoside through high-yield enzymic method - Google Patents
Preparation method for assisting in extracting stibene glucoside through high-yield enzymic method Download PDFInfo
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- CN105586378A CN105586378A CN201610144375.4A CN201610144375A CN105586378A CN 105586378 A CN105586378 A CN 105586378A CN 201610144375 A CN201610144375 A CN 201610144375A CN 105586378 A CN105586378 A CN 105586378A
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
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Abstract
The invention discloses a preparation method for assisting in extracting stibene glucoside through a high-yield enzymic method, and belongs to the technical field of preparation of stibene glucoside. Polygonum multiflorum is adopted as a raw material, high-pressure homogenization is firstly carried out, a composite enzyme agent is used for enzymolysis to increase the extraction rate, rotary steaming is carried out for concentration after microwave alcohol extraction, finally drying and grinding are carried out, and therefore the preparation method for assisting in extracting stibene glucoside through the high-yield enzymic method is obtained. The method is easy and convenient to operate, it is unnecessary to add a large amount of organic solution in the extracting process, organic matter residues are avoided, the operating environment is mild, and the problems of stibene glucoside oxidation and glucoside chain breakage caused by high temperature are avoided. Finally, the stibene glucoside extraction rate reaches 95% or above, the purity reaches 99% or above, and the preparation method is suitable for large-scale production application.
Description
Technical field
The preparation method who the invention discloses a kind of high yield enzyme process assisted extraction Stibene-glucoside, belongs to Stibene-glucoside preparing technical field.
Background technology
Stibene-glucoside claims again 2,3,5,4'-tetrahydroxyl diphenylethylene-2-O-BETA-D-glucoside, belongs to Polyhydroxystibene, and Stibene-glucoside is distinctive bioactive ingredients in fleece-flower root medicinal material, is the specificity index component of fleece-flower root medicinal material, soluble in water, methyl alcohol, ethanol. Pharmacological research shows, Stibene-glucoside have oxidation and removing free radicals, anti-ageing, reduce cholesterol, protect the liver and neuroprotection, can develop treatment senile dementia, reducing blood lipid and antiatherogenic novel drugs.
The existing method of preparing Stibene-glucoside that separates from the fleece-flower root: first adopting Diluted Alcohol is to extract solvent fleece-flower root medicinal material is carried out to refluxing extraction, then use ethyl acetate to extract extract, again ethyl acetate extract layer is carried out to silica gel column chromatography and SephadexLH-20 column chromatography for separation successively, finally prepare content and be 85% Stibene-glucoside. The major defect of the method is: 1. adopt high temperature reflux to extract Stibene-glucoside, and easily oxidized and glycosidic bond fracture of Stibene-glucoside under hot conditions, this extracting method damages Stibene-glucoside; 2. adopt normal pressure column chromatography separating for several times, loaded down with trivial details, the consuming time length of process, solvent consumption is large, has improved preparation cost; 3. component overstand and produce dead absorption in normal pressure pillar, sample losses is large, and the rate of recovery is lower.
In addition, at present artificial synthetic Stibene-glucoside has difficulty, and cost costliness, the Stibene-glucoside performance synthesizing is stable not, and purity is not high, at present from plant, extracting Stibene-glucoside is a kind of common method, but major part is extracted and also all adopted a large amount of organic solvents to extract from plant, not only complex operation is expensive, and the purity of the Stibene-glucoside making is still not high, and there is the problem of organic substance residues.
Therefore, make a kind of high-purity, the Stibene-glucoside of organic solvent-free is necessary thing.
Summary of the invention
The technical problem that the present invention mainly solves: for current conventional method extracting from plant in the process of Stibene-glucoside, all often at high temperature to carry out, make Stibene-glucoside easily oxidized and cause glycosidic linkage fracture, in addition in the time extracting, also adopt a large amount of organic solvents, not only complex operation, expensive, residual organic solvent also pollutes the environment, make the lower present situation of Stibene-glucoside purity making, provide a kind of taking the fleece-flower root as raw material, first through high-pressure homogeneous recycling complex enzyme agent enzymolysis to improve recovery rate, again by after microwave alcohol extracting, concentrated by rotary evaporation, final drying is ground, thereby obtain high yield enzyme process assisted extraction Stibene-glucoside. the method is easy and simple to handle, extract process in without add a large amount of organic solution, avoid organic substance residues, and operating environment gentleness, avoid high temperature to cause the problem of Stibene-glucoside oxidation and glycosidic linkage fracture, finally make Stibene-glucoside extraction rate reached more than 95%, purity, up to more than 99%, is applicable to large-scale production and application.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
(1) take after 1～2kg fleece-flower root is cleaned and dry and put into airslide disintegrating mill, with 3m3/ min air consumption and 0.70～0.80MPa air pressure are pulverized, to obtain fleece-flower root powder is that 1:2 pours in distilled water by solid-to-liquid ratio, be placed on magnetic stirrer, be uniformly mixed 10～20min with 200～300r/min rotating speed, obtain fleece-flower root powder suspension;
(2) fleece-flower root powder suspension obtained above is moved into high pressure homogenizer, carry out high-pressure homogeneous processing 5～8min with 2.0～3.0L/h flow velocity and 100～110MPa pressure, the fleece-flower root mixed liquor measuring after 1.0～1.2L homogeneous packs in the ceramic enzymatic vessel that volume is 2L, add 0.2～0.5% complex enzyme agent of enzymolysis substrate gross mass, be that 0.5mol/L acetum regulates pH to 4.0～5.0 by concentration, be placed in insulating box in 35～45 DEG C of lower seal enzymolysis 60～80min;
(3) enzymolysis complete after isolated by filtration remove filter residue, obtain enzymolysis liquid, enzymolysis liquid is packed in pipe type ultrafiltration device, utilize the CAM of 100,000 molecular weight as milipore filter, and carry out ultrafiltration under the condition of the driving pressure of 0.1～0.2MPa and 20～30 DEG C, collect filtrate;
(4) by the filtrate after above-mentioned ultrafiltration and absolute ethyl alcohol by volume for 1:1 mixes, put into Microwave Extraction instrument, with 200～250W power heating to 40～50 DEG C, insulation microwave alcohol extracting 1～2h, after alcohol extracting completes, mixed liquor is put into rotary evaporator, be heated to 45～55 DEG C, revolve steaming recovery absolute ethyl alcohol concentration until mixeding liquid volume becomes 1/8～1/6 of original volume, obtain concentrate;
(5) concentrate obtained above is put into vacuum freeze-drying machine, lower the temperature until condenser temperature reaches-40～-30 DEG C with 1 DEG C/min speed, freeze drying 30～40min, puts into agate mortar after being dried again and pulverizes and cross 200 mesh standard sieves, packaging, obtains Stibene-glucoside.
Complex enzyme agent described in step (2) is to be made for 2:1 is composite in mass ratio by cellulase and neutral proteinase.
Application process of the present invention is: the Stibene-glucoside powder that the present invention is made is made talan glycosides capsule, 1～3g Stibene-glucoside powder is wherein housed in each capsule, when edible, be equipped with 250～350mL warm water delivery service, take after the meal once every day three, each serving with 1～3, long-term use 10～15 days, record body's cholesterol and reduce, resistance is improved, and antidotal effect is remarkable.
The invention has the beneficial effects as follows: the method is easy and simple to handle, extract process in without add a large amount of organic solution, avoid organic substance residues, and operating environment gentleness, avoid high temperature to cause the problem of Stibene-glucoside oxidation and glycosidic linkage fracture, finally make Stibene-glucoside extraction rate reached more than 95%, purity, up to more than 99%, is applicable to large-scale production and application.
Detailed description of the invention
First take after 1～2kg fleece-flower root is cleaned and dry and put into airslide disintegrating mill, with 3m3/ min air consumption and 0.70～0.80MPa air pressure are pulverized, to obtain fleece-flower root powder is that 1:2 pours in distilled water by solid-to-liquid ratio, be placed on magnetic stirrer, be uniformly mixed 10～20min with 200～300r/min rotating speed, obtain fleece-flower root powder suspension; Then fleece-flower root powder suspension obtained above is moved into high pressure homogenizer, carry out high-pressure homogeneous processing 5～8min with 2.0～3.0L/h flow velocity and 100～110MPa pressure, the fleece-flower root mixed liquor measuring after 1.0～1.2L homogeneous packs in the ceramic enzymatic vessel that volume is 2L, add 0.2～0.5% complex enzyme agent of enzymolysis substrate gross mass, be that 0.5mol/L acetum regulates pH to 4.0～5.0 by concentration, be placed in insulating box in 35～45 DEG C of lower seal enzymolysis 60～80min; After enzymolysis completes, isolated by filtration is removed filter residue, obtain enzymolysis liquid, enzymolysis liquid is packed in pipe type ultrafiltration device, utilize the CAM of 100,000 molecular weight as milipore filter, and carry out ultrafiltration under the condition of the driving pressure of 0.1～0.2MPa and 20～30 DEG C, collect filtrate; Next by the filtrate after above-mentioned ultrafiltration and absolute ethyl alcohol by volume for 1:1 mixes, put into Microwave Extraction instrument, with 200～250W power heating to 40～50 DEG C, insulation microwave alcohol extracting 1～2h, after alcohol extracting completes, mixed liquor is put into rotary evaporator, be heated to 45～55 DEG C, revolve steaming recovery absolute ethyl alcohol concentration until mixeding liquid volume becomes 1/8～1/6 of original volume, obtain concentrate; Finally concentrate obtained above is put into vacuum freeze-drying machine, lower the temperature until condenser temperature reaches-40～-30 DEG C with 1 DEG C/min speed, freeze drying 30～40min, puts into agate mortar after being dried again and pulverizes and cross 200 mesh standard sieves, packaging, obtains Stibene-glucoside.
Example 1
First take after the 1kg fleece-flower root is cleaned and dry and put into airslide disintegrating mill, with 3m3/ min air consumption and 0.70MPa air pressure are pulverized, and will obtain fleece-flower root powder is that 1:2 pours in distilled water by solid-to-liquid ratio, is placed on magnetic stirrer, is uniformly mixed 10min with 200r/min rotating speed, obtains fleece-flower root powder suspension; Then fleece-flower root powder suspension obtained above is moved into high pressure homogenizer, carry out high-pressure homogeneous processing 5min with 2.0L/h flow velocity and 100MPa pressure, the fleece-flower root mixed liquor measuring after 1.0L homogeneous packs in the ceramic enzymatic vessel that volume is 2L, add the 0.2% complex enzyme agent of enzymolysis substrate gross mass, be that 0.5mol/L acetum regulates pH to 4.0 by concentration, be placed in insulating box in 35 DEG C of lower seal enzymolysis 60min; After enzymolysis completes, isolated by filtration is removed filter residue, obtains enzymolysis liquid, and enzymolysis liquid is packed in pipe type ultrafiltration device, utilizes the CAM of 100,000 molecular weight as milipore filter, and carries out ultrafiltration under the condition of the driving pressure of 0.1MPa and 20 DEG C, collects filtrate; Next by the filtrate after above-mentioned ultrafiltration and absolute ethyl alcohol by volume for 1:1 mixes, put into Microwave Extraction instrument, with 200W power heating to 40 DEG C, insulation microwave alcohol extracting 1h, after alcohol extracting completes, mixed liquor is put into rotary evaporator, be heated to 45 DEG C, revolve steaming recovery absolute ethyl alcohol concentration until mixeding liquid volume becomes 1/8 of original volume, obtain concentrate; Finally concentrate obtained above is put into vacuum freeze-drying machine, lower the temperature until condenser temperature reaches-40 DEG C with 1 DEG C/min speed, freeze drying 30min, puts into agate mortar after being dried again and pulverizes and cross 200 mesh standard sieves, packaging, obtains Stibene-glucoside.
This case method is unique novel, when use, the Stibene-glucoside powder that the present invention is made is made talan glycosides capsule, and 1g Stibene-glucoside powder is wherein housed in each capsule, when edible, be equipped with 250mL warm water delivery service, take after the meal once every day three, each serving with 1, long-term use 10 days, record body's cholesterol and reduce, resistance is improved, and antidotal effect is remarkable.
Example 2
First take after the 1.5kg fleece-flower root is cleaned and dry and put into airslide disintegrating mill, with 3m3/ min air consumption and 0.75MPa air pressure are pulverized, and will obtain fleece-flower root powder is that 1:2 pours in distilled water by solid-to-liquid ratio, is placed on magnetic stirrer, is uniformly mixed 15min with 250r/min rotating speed, obtains fleece-flower root powder suspension; Then fleece-flower root powder suspension obtained above is moved into high pressure homogenizer, carry out high-pressure homogeneous processing 7min with 2.5L/h flow velocity and 105MPa pressure, the fleece-flower root mixed liquor measuring after 1.1L homogeneous packs in the ceramic enzymatic vessel that volume is 2L, add the 0.35% complex enzyme agent of enzymolysis substrate gross mass, be that 0.5mol/L acetum regulates pH to 4.5 by concentration, be placed in insulating box in 40 DEG C of lower seal enzymolysis 70min; After enzymolysis completes, isolated by filtration is removed filter residue, obtains enzymolysis liquid, and enzymolysis liquid is packed in pipe type ultrafiltration device, utilizes the CAM of 100,000 molecular weight as milipore filter, and carries out ultrafiltration under the condition of the driving pressure of 0.15MPa and 25 DEG C, collects filtrate; Next by the filtrate after above-mentioned ultrafiltration and absolute ethyl alcohol by volume for 1:1 mixes, put into Microwave Extraction instrument, with 230W power heating to 45 DEG C, insulation microwave alcohol extracting 1.5h, after alcohol extracting completes, mixed liquor is put into rotary evaporator, be heated to 50 DEG C, revolve steaming recovery absolute ethyl alcohol concentration until mixeding liquid volume becomes 7/48 of original volume, obtain concentrate; Finally concentrate obtained above is put into vacuum freeze-drying machine, lower the temperature until condenser temperature reaches-35 DEG C with 1 DEG C/min speed, freeze drying 35min, puts into agate mortar after being dried again and pulverizes and cross 200 mesh standard sieves, packaging, obtains Stibene-glucoside.
This case method is unique novel, when use, the Stibene-glucoside powder that the present invention is made is made talan glycosides capsule, and 2g Stibene-glucoside powder is wherein housed in each capsule, when edible, be equipped with 300mL warm water delivery service, take after the meal once every day three, each serving with 2, long-term use 13 days, record body's cholesterol and reduce, resistance is improved, and antidotal effect is remarkable.
Example 3
First take after the 2kg fleece-flower root is cleaned and dry and put into airslide disintegrating mill, with 3m3/ min air consumption and 0.80MPa air pressure are pulverized, and will obtain fleece-flower root powder is that 1:2 pours in distilled water by solid-to-liquid ratio, is placed on magnetic stirrer, is uniformly mixed 20min with 300r/min rotating speed, obtains fleece-flower root powder suspension; Then fleece-flower root powder suspension obtained above is moved into high pressure homogenizer, carry out high-pressure homogeneous processing 8min with 3.0L/h flow velocity and 110MPa pressure, the fleece-flower root mixed liquor measuring after 1.2L homogeneous packs in the ceramic enzymatic vessel that volume is 2L, add the 0.5% complex enzyme agent of enzymolysis substrate gross mass, be that 0.5mol/L acetum regulates pH to 5.0 by concentration, be placed in insulating box in 45 DEG C of lower seal enzymolysis 80min; After enzymolysis completes, isolated by filtration is removed filter residue, obtains enzymolysis liquid, and enzymolysis liquid is packed in pipe type ultrafiltration device, utilizes the CAM of 100,000 molecular weight as milipore filter, and carries out ultrafiltration under the condition of the driving pressure of 0.2MPa and 30 DEG C, collects filtrate; Next by the filtrate after above-mentioned ultrafiltration and absolute ethyl alcohol by volume for 1:1 mixes, put into Microwave Extraction instrument, with 250W power heating to 50 DEG C, insulation microwave alcohol extracting 2h, after alcohol extracting completes, mixed liquor is put into rotary evaporator, be heated to 55 DEG C, revolve steaming recovery absolute ethyl alcohol concentration until mixeding liquid volume becomes 1/6 of original volume, obtain concentrate; Finally concentrate obtained above is put into vacuum freeze-drying machine, lower the temperature until condenser temperature reaches-30 DEG C with 1 DEG C/min speed, freeze drying 40min, puts into agate mortar after being dried again and pulverizes and cross 200 mesh standard sieves, packaging, obtains Stibene-glucoside.
This case method is unique novel, when use, the Stibene-glucoside powder that the present invention is made is made talan glycosides capsule, and 3g Stibene-glucoside powder is wherein housed in each capsule, when edible, be equipped with 350mL warm water delivery service, take after the meal once every day three, each serving with 3, long-term use 15 days, record body's cholesterol and reduce, resistance is improved, and antidotal effect is remarkable.
Claims (2)
1. a preparation method for high yield enzyme process assisted extraction Stibene-glucoside, is characterized in that concrete preparation process is:
(1) take after 1～2kg fleece-flower root is cleaned and dry and put into airslide disintegrating mill, with 3m3/ min air consumption and 0.70～0.80MPa air pressure are pulverized, to obtain fleece-flower root powder is that 1:2 pours in distilled water by solid-to-liquid ratio, be placed on magnetic stirrer, be uniformly mixed 10～20min with 200～300r/min rotating speed, obtain fleece-flower root powder suspension;
(2) fleece-flower root powder suspension obtained above is moved into high pressure homogenizer, carry out high-pressure homogeneous processing 5～8min with 2.0～3.0L/h flow velocity and 100～110MPa pressure, the fleece-flower root mixed liquor measuring after 1.0～1.2L homogeneous packs in the ceramic enzymatic vessel that volume is 2L, add 0.2～0.5% complex enzyme agent of enzymolysis substrate gross mass, be that 0.5mol/L acetum regulates pH to 4.0～5.0 by concentration, be placed in insulating box in 35～45 DEG C of lower seal enzymolysis 60～80min;
(3) enzymolysis complete after isolated by filtration remove filter residue, obtain enzymolysis liquid, enzymolysis liquid is packed in pipe type ultrafiltration device, utilize the CAM of 100,000 molecular weight as milipore filter, and carry out ultrafiltration under the condition of the driving pressure of 0.1～0.2MPa and 20～30 DEG C, collect filtrate;
(4) by the filtrate after above-mentioned ultrafiltration and absolute ethyl alcohol by volume for 1:1 mixes, put into Microwave Extraction instrument, with 200～250W power heating to 40～50 DEG C, insulation microwave alcohol extracting 1～2h, after alcohol extracting completes, mixed liquor is put into rotary evaporator, be heated to 45～55 DEG C, revolve steaming recovery absolute ethyl alcohol concentration until mixeding liquid volume becomes 1/8～1/6 of original volume, obtain concentrate;
(5) concentrate obtained above is put into vacuum freeze-drying machine, lower the temperature until condenser temperature reaches-40～-30 DEG C with 1 DEG C/min speed, freeze drying 30～40min, puts into agate mortar after being dried again and pulverizes and cross 200 mesh standard sieves, packaging, obtains Stibene-glucoside.
2. the preparation method of a kind of high yield enzyme process assisted extraction Stibene-glucoside according to claim 1, is characterized in that: the complex enzyme agent described in step (2) is to be made for 2:1 is composite in mass ratio by cellulase and neutral proteinase.
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| CN201610144375.4A CN105586378A (en) | 2016-03-15 | 2016-03-15 | Preparation method for assisting in extracting stibene glucoside through high-yield enzymic method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108208021A (en) * | 2018-03-02 | 2018-06-29 | 东莞市炫墨生物科技有限公司 | Biological bactericide and its purposes in daily chemical product |
| CN108245455A (en) * | 2018-03-02 | 2018-07-06 | 东莞市炫墨生物科技有限公司 | Loss preventing plant sends out agent and its purposes in daily chemical product |
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2016
- 2016-03-15 CN CN201610144375.4A patent/CN105586378A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108208021A (en) * | 2018-03-02 | 2018-06-29 | 东莞市炫墨生物科技有限公司 | Biological bactericide and its purposes in daily chemical product |
| CN108245455A (en) * | 2018-03-02 | 2018-07-06 | 东莞市炫墨生物科技有限公司 | Loss preventing plant sends out agent and its purposes in daily chemical product |
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