CN101082055A - Method for extracting flavonoids substance from liquorice slag by employing complex enzyme process - Google Patents
Method for extracting flavonoids substance from liquorice slag by employing complex enzyme process Download PDFInfo
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Abstract
The present invention discloses synthase process of extracting flavones from waste licorice residue. Waste licorice residue as material is treated through alcohol extraction, concentration, macroporous resin adsorption, organic solvent elution, collecting flavones, concentrating to recover solvent and drying to obtain licorice flavones product with licorice flavones content over 20 % and low impurity content. The process is simple and high in efficiency, and may have the available licorice processing apparatus utilized to saving investment.
Description
Technical field
The present invention relates to the method that a kind of licorice slag extracts Flavonoid substances, be specifically related to a kind of method of utilizing combined-enzyme method to extract Flavonoid substances from licorice slag.
Background technology
Radix Glycyrrhizae (Glycyrrhiza) is pulse family (Leguminosae) Papillionoideae (PapiliantaeTaub.) glycyrrhiza genus, is the extremely wide Chinese medicinal materials of a kind of application, have the title of " ten sides, nine grass ".Chinese scholars has been carried out many researchs to chemical ingredients and the pharmacological action of Radix Glycyrrhizae, thinks that its main effective constituent is Potenlini and flavonoid compound.Up to the present, very deep to the research of Potenlini both at home and abroad, now formed large-scale production.And the research of licoflavone is mainly concentrated in chemical ingredients and the pharmacological action, less to its extraction research.So in the production and exploitation of effective liquorice, flavones is discarded with the dregs of a decoction as submember, causes the significant wastage of resource.Along with further investigation to Radix Glycyrrhizae, people progressively recognize the importance of extracting these two kinds of materials from Radix Glycyrrhizae, and have done preliminary study, but these researchs are to utilize conventional extracting method mostly, time is long, cost is high, efficient is low, and these are all restricting the development of flavones suitability for industrialized production.
Licoflavone has good effect on industries such as medicine, healthcare products, makeup, foodstuff additive, the domestic and international market demand increases day by day.Many enterprise investments are planned to build licoflavone factory, but present most of Radix Glycyrrhizae processing enterprise is tradition processing, mostly be the thick products production of raw material level, scale and benefit are low, production level is poor, enterprise's extensive operation, and roughing production added value is low, main products is single, and product is limited to Radix Glycyrrhizae based articles such as Radix Glycyrrhizae section, licorice root ointment, Licorice, Potenlini powder more.Radix Glycyrrhizae is as a kind of important medicine resource, and too simple raw material processing be a kind of serious waste to resource, and important useful licoflavone class material composition abandons together along with licorice slag, all is a kind of heavy tremendous loss to enterprise, country beyond doubt.Face the big tide of 21 century natural product extract drugs, pharmacological research, the deep development of medicinal plant, application are the inevitable requirements of each herbal medicine processing enterprise existence.
Radix Glycyrrhizae is as the secondary natural crude drugs species of special-protection-by-the-State.Protection Radix Glycyrrhizae resource, rational exploitation and utilization, not only significant to existing fragile ecology, and the Sustainable development of national economy had certain promoter action.Licoflavone is with a wide range of applications at aspects such as medicine, healthcare products, makeup, foodstuff additive as the stronger composition of a class biological activity, and its market requirement is powerful day by day.Develop the research of extracting Flavonoid substances in extracting from glycyrrhiza residue, help the scientific utilization of resource, value added to improving the product pair, increasing peasant income etc. have great importance.
Traditional flavones extracting method has water seaoning, soda acid extraction, alcohol extracting method etc., and it utilizes the simple physics method to extract flavones in the Radix Glycyrrhizae, and efficient is low and easily cause environmental pollution.And utilize microwave, ultrasonic-assisted extraction method, supercritical CO recently
2Novel physical apparatus such as collection formulation etc. extracts the method for flavones, is subjected to extensive concern because of extraction efficiency is higher, but owing to have in production application that one-time investment is excessive, operation is loaded down with trivial details, makes it be subjected to a certain degree restriction in industrial application.
In the face of problem and the shortcoming that above-mentioned the whole bag of tricks exists, the various countries scientific worker begins to consider to utilize enzyme process assisted extraction Flavonoid substances.Utilize total flavones research in the Howthorn Leaf of Enzymatic Extraction as people such as Wang Xiao, Li Linbo; The Enzymatic Extraction technical study of Wang Hui, the good ginkgolic flavone glycoside of Liu Jia finds that it carried out the enzyme pre-treatment to raw material before the pure flooding of routine, can improve the total flavones yield greatly, and is simple.But above research is only set about from pre-treatment, lacks the follow-up research work that matches, and still for slightly carrying product, product impurity is more for the flavones product that obtains; And being added on when improving yield of enzyme also must increase albumen foreign matter content in the product, and then reduced the grade of product.
More in order to solve impurity in products, the insufficient problem of purifying process.People utilize methods such as absorbent charcoal method, silica gel column chromatography, Amberlyst process to carry out the extraction purifying of flavones product.But purifying is all low because of adsorption rate, yield is low or the regeneration difficulty owing to absorbent charcoal method, silica gel chromatography extract, and is eliminated day by day.And macroporous adsorbent resin does not contain the reticulated structure of ion-exchange group as a class that grew up in 20th century, has the good adsorption performance, and desorption is easy, and physical strength is good, can use repeatedly and advantages such as fluid resistance is little.Macroporous adsorbent resin had applied research more widely at aspects such as the extraction of Natural Medicine Chemistry composition, separation and purification, preparation process reforms in recent years.As the new technology of an extraction separation purifying, obtained very big attention.
Those skilled in the art know, and the Flavonoid substances in the glycyrrhiza residue mostly is crosslinked or is adsorbed on the cell walls, and traditional treatment process is difficult for opening this structure, or destroy licoflavone easily under condition extremely.How to make full use of efficient, the environmental protection characteristics of Enzymatic Extraction, utilize the advantage of macroporous resin adsorption purifying simultaneously, overcome the former because of the albumen impurity that adds enzyme and produce and slightly carry other impurity in the product, and the licoricon superior product quality of gained, the extraction yield height is significant, and comprehensive like this technical scheme is demanded in market urgently.
Summary of the invention
Mostly be crosslinked or be adsorbed on the cell walls at the Flavonoid substances in the present both at home and abroad glycyrrhiza residue, traditional treatment process is difficult for opening this structure, or destroys the present situation of licoflavone under condition extremely easily.The present invention makes full use of efficient, the environmental protection characteristics of Enzymatic Extraction, utilize the advantage of macroporous resin adsorption purifying simultaneously, setting up a whole set of utilizes combined-enzyme method to extract the method for Flavonoid substances from licorice slag, overcome prior art because of the albumen impurity that adds enzyme and produce and slightly carry other impurity in the product, the licoricon superior product quality that obtains, the extraction yield height.
The invention provides a kind of method of utilizing combined-enzyme method to extract Flavonoid substances from licorice slag.The concrete grammar step is as follows:
1. preparation glycyrrhiza residue extracting solution, by with chloroform: formic acid: methyl alcohol=5: 0.5: 1 is developing agent, is developer with 8%AlCl3, carries out the silica gel thin sheet chromatographic analysis, determines licoflavone content;
With neutral cellulase and polygalacturonase prozyme under the phosphoric acid buffer of 5mMol/L, pH6.0,55 ℃ of effect 2h handle the glycyrrhiza residue extracting solution, wherein, the suitableeest enzyme of neutral cellulase is lived and is PH4.5-6.3, optimum temperuture is 55 ℃, and addition is 50U/ml, and the suitableeest enzyme of polygalacturonase is lived and is PH4.0-6.1, optimum temperuture is 57 ℃, and addition is 100U/ml;
3. extracting solution is added dehydrated alcohol, adopting solid-liquid ratio is 1: 14, is mixed with 80% ethanolic soln, boils 1h, and extraction time is 40min, carries out alcohol extracting, concentrates to reclaim solvent;
4. the employing rate of expansion is 1.67 SP825 resin, and its adsorption rate is 91.04 (mg/g), the glycyrrhiza residue extracting solution after absorption concentrates under the pH4.0-8.0 condition;
5. select 1: 15 chromatography column of aspect ratio for use, by ethanol and water alternate repetition wash-out, adopting ethyl acetate is best eluting solvent, and consumption is 2-3 a times of resin volume, under 10-50 ℃ static conditions, elution flow rate is 10ml/h, crosses the concentration 2% of post liquid, and liquid volume added is a column volume 1/3, the application of sample flow velocity is 20ml/h, alternately wash-out is 2-3 time, keeps resin isolation to use preceding state, collects the total flavones material simultaneously.
It is raw material with the licorice slag that the present invention specifically provides a kind of, by the processing of prozyme system, through ethanol-extracted, will concentrate back extracting solution absorption with macroporous adsorbent resin again, the method for collecting total flavones through the organic solution wash-out.
1, the extracting solution that glycyrrhiza residue is handled gained has carried out the thin plate silica gel column chromatography, by determining the major ingredient of extracting solution with the check analysis of standard specimen.
The present invention can pass through with chloroform: formic acid: methyl alcohol was developing agent according to 5: 0.5: 1, and the silica gel thin sheet chromatography is a developer with 8%AlCl3, and utilizing Flavonoid substances and AlCl3 to act on has special green fluorescence under the UV-light, determined that licoflavone exists.
2, prozyme pretreatment system reaction conditions determines.Determine the suitableeest enzyme of neutral cellulase PH4.5-6.3 alive, 55 ℃ of optimum temperutures, the suitableeest enzyme of polygalacturonase PH4.0-6.1 alive, 57 ℃ of optimum temperutures, the neutral cellulase addition is 50U/ml, and polygalacturonase 100U/ml is under the phosphoric acid buffer of 5mMol/L, pH6.0,55 ℃ of effect 2h, obvious to the licoflavone extraction effect.
3, the pretreated alcohol extracting system of prozyme determines.The parameter of alcohol extracting is the enzyme pretreatment fluid to be added dehydrated alcohol be mixed with 80% ethanolic soln, and solid-liquid ratio is 1: 14, and boiling the alcohol extracting time is 40min, crosses the elimination filter residue, utilizes colorimetric method for determining licoflavone extracted amount.(with the rutin is standard reference material, adopts spectrophotometry 510nm to measure OD, and the drawing standard curve gets equation: Y=73.00A-0.03003, and the unit of Y is mg/L)
4, select the SP825 of Mitsubishi chemical company for use, determined the parameter of resin.Resin is 1.67 through pretreated rate of expansion, adsorption rate is 91.04 (mg/g), at pH4.0-8.0, by ethanol and water alternate repetition wash-out, can remove the residue in the resin, the eluting solvent consumption is 2-3 a times of resin volume, and alternately wash-out is 2-3 time, finally, use preceding state before keeping separating with behind the water elution.Regeneration of resin is used the repeated treatments of methyl alcohol, acetone or soda acid, water, can recover the adsorptive power of resin.
5, the selection of elution requirement and macroporous resin column chromatography condition determines.1: 15 chromatography column of aspect ratio has been selected in the present invention's design for use, determines the concentration 2% of post liquid, and liquid volume added is a column volume 1/3, and flow velocity is 20ml/h.With the ethyl acetate is eluent, and the control elution flow rate is 10ml/h, elute effect the best.
By above technological invention, can reach following technique effect:
This method realizes, with the waste residue behind the crude extracts such as Radix Glycyrrhizae or Radix Glycyrrhizae tradition processing extraction Radix Glycyrrhizae extractum is raw material, after handling, the prozyme system carries out ethanol-extracted, concentrate the back absorption with macroporous adsorbent resin, collect total flavones through the organic solution wash-out, its gained Radix Glycyrrhizae total flavones content reaches more than 20%, and foreign matter content is low, and concentrating the recovery solvent can use repeatedly.
Description of drawings
1. utilize spectrophotometer scanning quantitative analysis to measure total flavonoids substance in the glycyrrhiza residue.
2. the effect of different methods pre-treatment licoflavone class material, the concrete implication of numeral is among the figure, and 1 be burning water, and 2 be alcohol extracting method, and 3 is ultrasonic pretreatment, and 4 is cellulose treatment, and 5 is the polygalacturonase processing, and 6 is the highly basic processing.
3. the influence of different concns polygalacturonase addition to extracting.
4. the prozyme system is handled the proportion relation of Flavonoid substances.
5. the influence of enzyme pretreatment temperature to extracting.
6. the influence of enzyme pre-treatment pH to extracting.
7. the influence of enzyme pretreatment time to extracting.
8. the influence of different feed liquid comparison extraction.
9. the influence of different ethanol concentration to extracting.
10. with the ethyl acetate eluent, to the influence of PH to desorb.
11. with the ethyl acetate is eluent, to the influence of temperature to desorb.
12. extract concentration is to the influence of chromatography column absorption.
13. application of sample speed is to the influence of chromatography column absorption.
14. determining of elution flow rate.
Embodiment
Below, describe the present invention in detail by reference example, still, what those having ordinary skill in the art will appreciate that is that the present invention is not limited to following embodiment.In addition, in following explanation, if no special instructions, then % all refers to weight percent.
Embodiment one: qualitative, the quantitative assay of Flavonoid substances in the glycyrrhiza residue
Qualitative minute property: with chloroform: formic acid: methyl alcohol=5: 0.5: 1 is developing agent, and the silica gel thin sheet chromatography is a developer with 8%AlCl3, and utilizing Flavonoid substances and AlCl3 to act on has special green fluorescence under the UV-light, determine that licoflavone exists.
Quantitative analysis: with the rutin is standard substance, the an amount of rutin that draws the 1ml80% dissolve with ethanol adds in the volumetric flask of 10ml, adding 0.4ml 5%NaNO2 shakes and all places 6Min, adding 0.4ml 10%Al (NO3) shakes and all places 6Min, add 4ml 4%NaOH again, adding 80% ethanol is settled to 10ml and shakes and all place 15Min, scan with scanning spectrophotometer, as seen its 350,505nm respectively has a peak, wherein 505 places are the peculiar absorption peak of Flavonoid substances, therefore determine that the optimum determining wavelength is 505nm, referring to accompanying drawing 1.
Embodiment two: single enzyme and prozyme system are handled the simultaneous test of licorice slag extracting solution reaction conditions
The existing technical grade generally the suitableeest enzyme of enzyme-cellulose enzyme PH4.5-7.8 alive, 65 ℃ of the highest operative temperatures, the suitableeest enzyme of polygalacturonase PH4.0-7.3 alive, 63 ℃.The suitableeest enzyme of the neutral cellulase of selecting for use Ningxia jade of the He family Bioisystech Co., Ltd to produce PH4.5-6.3 alive, 55 ℃ of optimum temperutures, the suitableeest enzyme of polygalacturonase PH4.0-6.1 alive, 57 ℃, and there is higher lignoenzyme enzyme to live in its cellulase, meet the present invention's needs, therefore determine cellulase, polygalacturonase.The present invention simultaneously handles in above neutral cellulase and the polygalacturonase parameter selected for use according to selecting for use different manufacturers neutral cellulase and polygalacturonase to handle, and the effect of the licoflavone of its extraction is identical.
After selecting the diverse ways pre-treatment for use, other condition is identical, behind the effect 1h, extracting solution is added dehydrated alcohol be mixed with 80% ethanolic soln, measuring the licoflavone extracted amount learns, select for use prozyme to handle glycyrrhiza residue, remarkable for Flavonoid substances effect crosslinked in effective extraction glycyrrhiza residue or that be adsorbed on the cell walls.It is good than other treatment process to add cellulose treatment and polygalacturonase treatment effect as can be seen by accompanying drawing 2 in above the whole bag of tricks, and wherein the polygalacturonase effect is more outstanding, therefore selects for use cellulose treatment and polygalacturonase as pretreatment process.
Embodiment three: the prozyme system is handled determining of glycyrrhiza residue extracting solution condition
Adopt the suitableeest enzyme of neutral cellulase PH4.5-6.3 alive, 55 ℃ of optimum temperutures, the suitableeest enzyme of polygalacturonase PH4.0-6.1 alive, 57 ℃, consider the influence of PH to the licoflavone structural stability, consider the influence of salt concn simultaneously to the back extraction separation.Therefore the preferential selective action condition of the present invention is under the phosphoric acid buffer of 5mMol/L, pH6.0,55 ℃ of processing.
The effect that is added cellulase and polygalacturonase processing by accompanying drawing 2 as can be seen in above the whole bag of tricks is good than other treatment process, and wherein the polygalacturonase effect is more outstanding, therefore selects for use cellulose treatment and polygalacturonase as pretreatment process.
Almost reach maximum value by different concns polygalacturonase addition to extracting influence as can be seen when addition is 100U/ml, extracting, when continuing to increase concentration, it is little to extract influence, and it is 100U/ml that the consideration cost is selected the polygalacturonase addition for use, referring to accompanying drawing 3.
With the polygalacturonase addition is 100U/ml, adds the enzyme of different ratios respectively, the different time sampling.Add dehydrated alcohol and make 80% ethanolic soln, boil 1h, measure the licoflavone extracted amount in sample ligand.
Can see, extracted amount reaches maximum value substantially when being 0.5: 1 when cellulase and polygalacturonase add, though continue the addition extracted amount of increased fiber element certain increase is arranged, but DeGrain, consideration cost problem was determined to select for use 0.5: 1, be that the cellulase addition is 50U/ml, polygalacturonase 100U/ml is referring to accompanying drawing 4.
Show that by the repeated multiple times experiment the present invention determines that finally the neutral cellulase addition is 50U/ml, polygalacturonase 100U/ml, under the phosphoric acid buffer of 5mMol/L, pH6.0,55 ℃ of effect 2h are referring to accompanying drawing 5, accompanying drawing 6, accompanying drawing 7.
Embodiment four: the determining of alcohol extracting condition
The present invention makes 80% ethanolic soln by adding dehydrated alcohol in sample ligand, boils 1h, measures the licoflavone extracted amount.
Experiment shows that the condition of glycyrrhiza residue alcohol extracting is that alcohol concn is 80%, adopts and determines that solid-liquid ratio is 1: 14, and extraction time is 40min, and the extraction yield of flavones can reach about 2.0% in the glycyrrhiza residue.Referring to accompanying drawing 8, accompanying drawing 9.
Embodiment five: the static conditions of macroporous resin is determined
1. the type selecting of macroporous resin: the SP825 that has selected Mitsubishi Chemical Ind for use.
2. the rate of expansion of resin: get 50 ℃, the resin 1.12g (2.4ml) of the oven dry of spending the night adds alcohol-pickledly, and its volume change is 4.0ml.Rate of expansion=4.0/2.4=1.67
3. the processing of resin:
Pre-treatment by ethanol and water alternate repetition wash-out, can be removed the residue in the resin, and the 2-3 that general eluting solvent consumption is the resin volume doubly replaces wash-out 2-3 time, finally with behind the water elution, uses preceding state before keeping separating.
Regeneration of resin is used the repeated treatments of methyl alcohol, acetone or soda acid, water, can recover the adsorptive power of resin
4. the adsorption rate of resin:
Take by weighing resin 0.4g, add the static absorption of licorice extract 5ml 20 hours, intermittently with hand moving.
Adsorption rate=Δ C * V/m=7.34 * 5/0.4=91.04 (mg/g)
| 1 | 2 | 3 | |
| C0(mg/ml) Ct(mg/ml) ΔC(mg/ml) | 11.65 4.31 7.34 | 11.62 4.23 7.39 | 11.59 4.24 7.34 |
Wherein: at the beginning of the C0-extracting solution, after the Ct-absorption, Δ C=C0-Ct
5. the influence of extract concentration to adsorbing:
Take by weighing resin 0.4g, add the static absorption of licorice extract (different extension rate) 5ml 20 hours, intermittently with hand moving.
| Concentration (mg/ml) | 11.5 | 9.2 | 6.9 | 4.6 | 2.3 | 0.057 |
| Adsorptive capacity (mg) | 31.5 | 21.2 | 18.5 | 10.2 | 4.3 | 1.2 |
As can be seen under static conditions, extracting solution flavones concentration high resin adsorptive capacity more is high more by table.
6. the influence of extracting solution pH to adsorbing:
Take by weighing resin 0.4g, add licorice extract 4ml (starting point concentration 12.6mg/ml) and regulate different pH, and add to 5ml with ethanol, static absorption 20 hours is intermittently with hand moving.
| pH | 4.0 | 5.0 | 6.0 | 7.0 | 8.0 |
| Adsorptive capacity (mg) | 24.5 | 24.7 | 25.0 | 25.1 | 24.9 |
By table as can be seen, extracting solution influences little to adsorption rate and the one-tenth irregular fluctuation at pH4.0-8.0.
Embodiment six: the determining of elution requirement
1. the selection of eluent:
Absorption is (adsorptive capacity is 26.7mg) as previously mentioned.With filter paper elimination supernatant liquor, use the different methyl alcohol of polarity, ethanol, acetone, ethyl acetate to make eluent respectively.Survey the wash-out yield.
| Solvent | Methyl alcohol | Ethanol | Acetone | Ethyl acetate |
| Elution amount (mg) eluting rate (%) | 10.27 38.5 | 10.81 40.5 | 14.5 53.4 | 17.59 65.3 |
Experiment finds that the ethyl acetate effect is best, and it is former because licoflavone is soluble in the less reagent of polarity, so easier wash-out.
2. with the ethyl acetate eluent, to the influence of PH and temperature to desorb
By accompanying drawing 10, accompanying drawing 11 as can be seen, elutriant pH is little to the wash-out influence.In the increase of 10-50 ℃ of following desorption efficiency along with temperature, efficient increases, but desorption effect obviously descends after having crossed 50 ℃, shows that the molecular motion of temperature rising at a certain temperature strengthens, and the solubleness increase of licoflavone helps desorb.And may be unfavorable for desorb because of variation and other condition influence of resin internal structure after reaching certain temperature.
3. the comparison of different resins performance
| Model | Equilibrium concentration (mg/l) | Adsorption rate (mg/g) | Desorption efficiency (%) |
| |
35 | 95.1 | 9 3 |
| ADS97 | 1020 | 70 | 78.2 |
| DM130 | 800 | 30.2 | 95 |
As can be seen from the above table, extract the macroporous resin of ADS97A, DM130 commonly used as Flavonoid substances and compare with SP825, its every performance is all low than the latter, thus this research to select SP825 for use be correct.
Embodiment seven: the determining of macroporous resin column chromatography condition
According to resin chromatography column claimed range 1: 5-1: between 20, consider adsorption volume, separating effect and practical application, 1: 15 chromatography column of blade diameter length ratio has been selected in design for use voluntarily.
Determine the influence of extract concentration coupled columns absorption.With the liquid volume added is column volume 1/3, and flow velocity is the 5ml/h application of sample.With 1%, 2%, 3%, 4% concentration application of sample, measure the concentration of its effluent liquid respectively, its result referring to accompanying drawing 12 as can be seen, along with the increase of sample concentration, it oozes dew point in advance.But the too small whole work efficiency that can not bring into play chromatography column of concentration, taking all factors into consideration selection is upper prop concentration with about 2%.
When adopting ethyl acetate to be eluent, respectively with 5,10,15,20ml/h measures the concentration of its effluent liquid and elution volume.
Determine the influence of application of sample speed coupled columns absorption.With 2% licoflavone solution is upper prop concentration, respectively with 5,10,15,20, the last sample speed of 25ml/h, measure the concentration of its effluent liquid.By accompanying drawing 13 as can be seen, it oozes dew point in advance along with the increase of flow velocity, and its reason may be unfavorable for the absorption fully of resin for flow velocity increases, thereby causes oozing dew point in advance.But variation is slower relatively below 20ml/h, takes all factors into consideration and determines with 20ml/h to be application of sample speed.
To increase its elution volume bigger along with the suction speed of taking off as can be seen by accompanying drawing 14, and its concentration is on the low side, and this certainly will make troubles for the post-treatment of licoflavone.Take all factors into consideration and determine that elution flow rate is 10ml/h.
Claims (1)
1, the invention provides a kind of method of utilizing combined-enzyme method from licorice slag extraction Flavonoid substances, its method steps is as follows,
(1). preparation glycyrrhiza residue extracting solution, by with chloroform: formic acid: methyl alcohol=5: 0.5: 1 is developing agent, is developer with 8%AlCl3, carries out the silica gel thin sheet chromatographic analysis, determines licoflavone content;
(2). with neutral cellulase and polygalacturonase prozyme under the phosphoric acid buffer of 5mMol/L, pH6.0,55 ℃ of effect 2h handle the glycyrrhiza residue extracting solution, wherein, the suitableeest enzyme of neutral cellulase is lived and is PH4.5-6.3, optimum temperuture is 55 ℃, and addition is 50U/ml, and the suitableeest enzyme of polygalacturonase is lived and is PH4.0-6.1, optimum temperuture is 57 ℃, and addition is 100U/ml;
(3). extracting solution is added dehydrated alcohol, and adopting solid-liquid ratio is 1: 14, is mixed with 80% ethanolic soln, boils 1h, and extraction time is 40min, carries out alcohol extracting, concentrates to reclaim solvent;
(4). the employing rate of expansion is 1.67 SP825 resin, and its adsorption rate is 91.04 (mg/g), the glycyrrhiza residue extracting solution after absorption concentrates under the pH4.0-8.0 condition;
(5). select 1: 15 chromatography column of aspect ratio for use, by ethanol and water alternate repetition wash-out, adopting ethyl acetate is best eluting solvent, and consumption is 2-3 a times of resin volume, under 10-50 ℃ static conditions, elution flow rate is 10ml/h, crosses the concentration 2% of post liquid, and liquid volume added is a column volume 1/3, the application of sample flow velocity is 20ml/h, alternately wash-out is 2-3 time, keeps resin isolation to use preceding state, collects the total flavones material simultaneously.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012185A (en) * | 2016-01-28 | 2017-08-04 | 亿利资源集团有限公司 | The preparation method and composite microporous separator of radix glycyrrhizae cellulose |
| CN107964000A (en) * | 2017-11-28 | 2018-04-27 | 张夏洋 | The double assisted extraction beeswax flavones of enzyme-ultrasound |
| CN108244295A (en) * | 2018-03-23 | 2018-07-06 | 青海康普生物科技股份有限公司 | A kind of preparation method of sea-buckthorn Fu tea |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1060385C (en) * | 1997-08-08 | 2001-01-10 | 中国科学院新疆化学研究所 | Licorice total flavone extracting method |
| CN1203068C (en) * | 2003-07-11 | 2005-05-25 | 上海奥利实业有限公司 | Extraction and purification method of licoflavone |
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- 2006-05-31 CN CNB2006100849589A patent/CN100529095C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012185A (en) * | 2016-01-28 | 2017-08-04 | 亿利资源集团有限公司 | The preparation method and composite microporous separator of radix glycyrrhizae cellulose |
| CN107964000A (en) * | 2017-11-28 | 2018-04-27 | 张夏洋 | The double assisted extraction beeswax flavones of enzyme-ultrasound |
| CN108244295A (en) * | 2018-03-23 | 2018-07-06 | 青海康普生物科技股份有限公司 | A kind of preparation method of sea-buckthorn Fu tea |
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