CN102633895A - Extraction and preparation method by comprehensively utilizing liquorice - Google Patents
Extraction and preparation method by comprehensively utilizing liquorice Download PDFInfo
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Abstract
The invention discloses an extraction and preparation method by comprehensively utilizing liquorice and belongs to the technical field of traditional Chinese medicine. Isoliquiritigenin, glycyrrhizic acid and liquorice polysaccharide which have high additional values are separated and prepared by a polyamide-macroporous resin coupling technique. The extraction and preparation method has the advantages of simple technical route and green and safe production process, does not need to use toxic and organic solvents and is suitable for mass production, the comprehensive utilization degree of the liquorice is greatly improved, the traditional Chinese medicine resource is saved, and good social benefit and economic benefit are generated.
Description
Technical field
The invention belongs to technical field of traditional Chinese medicines, be specifically related to a kind of preparation method who from Radix Glycyrrhizae, extracts preparation licorice polysaccharide, isoliquiritigenin and Potenlini simultaneously.
Background technology
Radix Glycyrrhizae is root and the rhizome of pulse family (Leguminosae) Glycyrrhiza (Glycyrrhiza) plant; The whole world has 29 kind of 6 mutation; Wherein China produces 18 kinds and 3 mutation, mainly is distributed in North China (comprising the Inner Mongol, Hebei) and northwest (comprising Ningxia, Gansu, Xinjiang, Qinghai, Shaanxi).As a kind of medicinal and ecological vegetable, the Radix Glycyrrhizae resource is more and more valuable, and therefore the comprehensive utilization to Radix Glycyrrhizae seems particularly important.
Modern chemistry research shows that mainly contain flavonoid glycoside composition (liquirtin, isoliquiritin etc.) and Potenlini constituents in the Radix Glycyrrhizae, wherein the content with liquirtin (content 0.5-1.5% in the medicinal material) and Potenlini (content 1.5-5% in the medicinal material) is the highest again.Also contain the Flavone aglycone constituents that polarity is less, content is very low (like Liquiritigenin, isoliquiritigenin, licochalcone A etc.) in addition in the Radix Glycyrrhizae, and the polyose composition.
Existing research shows, Potenlini has anti-inflammatory, antiviral, anti-allergic and immunoregulatory effect, is paid close attention to by Chinese scholars always; And licoflavone also found to have pharmacologically active widely in recent years, and wherein isoliquiritigenin is very remarkable in aspect effects such as anti-oxidant, antitumor and arrhythmias, and its drug effect is better than liquirtin, isoliquiritin etc., and added value of product is higher; Licorice polysaccharide has immunocompetence, can be widely used in food service industry.
At present; Existing patent " method of a kind of extraction separation Potenlini, licoflavone and licorice polysaccharide " (ZL 200610018275.3); Disclose a kind of simultaneous extraction and separated the method for Potenlini, licoflavone and licorice polysaccharide; After adopting solvent method to extract, carry out the preparation of the multiple composition of Radix Glycyrrhizae through methods such as organic solvent extraction, alcohol precipitation, acid are heavy again, but because licoflavone glycosides and Potenlini are the middle polarity composition; When adopting propyl carbinol, ETHYLE ACETATE, Pentyl alcohol, chloroform or primary isoamyl alcohol extracting licoflavone; Can not well licoflavone glycosides and Potenlini be separated, thereby make that the licoflavone (the UV method measures about 50%) for preparing among the embodiment and the content of Potenlini (HPLC measures about 35%) all are not very high, and more than the use of poisonous organic solvent all can bring dissolvent residual and influence the quality of extract.
Patent " comprehensive extraction method of glycyrrhiza " (application number 200810187964.6) also discloses a kind of comprehensive extraction method of glycyrrhiza; Licorice medicinal materials adopts the Hydrocerol A of 8-12% to extract licorice polysaccharide successively; With the ammonia extraction Potenlini of pH7-8, again with the sodium hydroxide extraction licoflavone of pH11-12.But in fact, the polarity of licoflavone glycosides and Potenlini is approaching, and the aqueous solution of different pH is relatively poor to the specificity that all types of compositions of Radix Glycyrrhizae extract; Each position content that extracts preparation is lower; About 30% like glycyrrhizic acid content, licoflavone content is about 50%, licorice polysaccharide only 19%.
Patent " is extracted the method for purifying isoliquiritigenin " from Radix Glycyrrhizae (application number: 201110287837.5) with " from Radix Glycyrrhizae, extracting the method for isoliquiritigenin " (application number: 200810072931.7) also disclose the method that from Radix Glycyrrhizae extraction prepares isoliquiritigenin; But above two kinds of preparing methods; All adopt earlier the licorice extract acid hydrolysis; Alkali open ring, repurity separate the acquisition isoliquiritigenin.The same problem that these two kinds of schemes are brought is: 1. owing to contain licoflavone (being mainly liquirtin) and Potenlini constituents in the licorice extract; Thereby Potenlini also can be hydrolyzed the generation glycyrrhetinic acid in licoflavone acid hydrolysis process, and this makes the Potenlini composition be destroyed and can't utilize; 2. the glycyrrhetinic acid polarity that after acid hydrolysis, generates of Potenlini is lower; Very approaching with the polarity of isoliquiritigenin; Therefore; Often will use normal-phase chromatographies such as silica gel column chromatography at the separation and purification isoliquiritigenin, when increasing production cost greatly, the use of poisonous organic solvent brings influence also for the quality of isoliquiritigenin product.
Therefore, in the extraction preparation method of comprehensive utilization Radix Glycyrrhizae, it is vital how adopting simple and effective stripping technique to separate to contained licoflavone, Potenlini and polyose composition in the licorice medicinal materials.
Summary of the invention
Technical problem to be solved by this invention provides a kind of extraction preparation method who fully utilizes Radix Glycyrrhizae, and this operational path is green, simple, is suitable for large-scale production, and the content of prepared isoliquiritigenin, Potenlini and licorice polysaccharide is higher.
In order to solve the problems of the technologies described above, the technical scheme that the present invention adopts is following:
A kind of extraction preparation method who fully utilizes Radix Glycyrrhizae is characterized in that it comprises the steps:
A. extracting liquorice medicinal material through 0～50% extraction using alcohol, reclaim under reduced pressure extracting solution, adds water again and makes in the soup every milliliter to contain crude drug amount 0.1～0.2g, regulates soup pH to 6～7, successively through polymeric amide chromatographic column and macroporous resin chromatography;
B. licorice polysaccharide preparation: collect through the effluent behind the macroporous resin, specific density is 1.10～1.20 when being evaporated to 65 ℃, and adding ethanol to determining alcohol is 80%, places, and separates out deposition, filters, and drying promptly gets licorice polysaccharide;
C. isoliquiritigenin preparation: the polymeric amide chromatographic column with 20～40% ethanol elutions of 1～3 times of column volume, is collected ethanol eluate more earlier with 1～3 times of column volume deionized water wash-out, and decompression recycling ethanol is placed and separated out the liquirtin deposition, filters; The extracting liquorice glycosides is with 40～60% dissolve with ethanol, adds hydrochloric acid again and makes that concentration of hydrochloric acid is 0.5～2mol/L in the solution, and 80～100 ℃ of acid hydrolysiss 2 hours are put cold; Hydro-oxidation sodium is regulated pH to neutral, continues hydro-oxidation sodium again and makes that naoh concentration is 1～3mol/L in the solution, and 80～100 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 2～5 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying promptly gets isoliquiritigenin again;
D. Potenlini preparation: macroporous resin chromatography with 60～80% ethanol elutions of 2～6 times of column volumes, is collected ethanol eluate more earlier with 1～3 times of column volume deionized water wash-out; Decompression recycling ethanol, glycyrrhizic aqueous solution is regulated pH to 2～3 with hydrochloric acid, places; Separate out deposition; Filter, drying promptly gets Potenlini.
The extraction preparation method of above-described comprehensive utilization Radix Glycyrrhizae is characterized in that the specification of used polymeric amide is the 80-100 order, with the weight ratio of licorice medicinal materials be 2: 1～4; The model of big pore resin is AB-8 or HPD-400, with the weight ratio of licorice medicinal materials be 2: 1～4.
The extraction preparation method of above-described comprehensive utilization Radix Glycyrrhizae; It is characterized in that the isoliquiritigenin preparation method is: the polymeric amide chromatographic column is earlier with 2 times of column volume deionized water wash-outs, again with 30% ethanol elution of 2 times of column volumes; Collect ethanol eluate; Decompression recycling ethanol is placed and is separated out the liquirtin deposition, filters; The extracting liquorice glycosides is with 50% dissolve with ethanol, adds hydrochloric acid again and makes that concentration of hydrochloric acid is 1mol/L in the solution, and 90 ℃ of acid hydrolysiss 2 hours are put cold; Regulate pH to neutral, continue hydro-oxidation sodium again and make that naoh concentration is 2mol/L in the solution, 90 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 3 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying promptly gets isoliquiritigenin again.
The extraction preparation method of above-described comprehensive utilization Radix Glycyrrhizae is characterized in that the preparation method of Potenlini is: macroporous resin chromatography is earlier with 2 times of column volume deionized water wash-outs; With 70% ethanol elution of 4 times of column volumes, collect ethanol eluate, decompression recycling ethanol again; Soup is regulated pH to 2, places, and separates out deposition; Filter, drying promptly gets Potenlini.
The extraction preparation method of above-described comprehensive utilization Radix Glycyrrhizae, it is characterized in that the condition of said decompression recycling ethanol is: pressure is 0.07～0.12MPa, temperature is 60～80 ℃.
The extraction preparation method of above-described comprehensive utilization Radix Glycyrrhizae is characterized in that the weight ratio of polymeric amide and licorice medicinal materials is 1: 1; The weight ratio of macroporous resin and licorice medicinal materials is 1: 1.
Beneficial effect: the present invention compared with prior art has following advantage:
(1) characteristics of the present invention are to have taken into full account the contained chemical ingredients type feature of Radix Glycyrrhizae; The character of utilizing polymeric amide all can not adsorb licorice polysaccharide the characterization of adsorption and the resin of Potenlini constituents to the specificity of flavones absorption, macroporous resin; Set up Radix Glycyrrhizae comprehensive utilization extraction separation scheme; Avoided the use of poisonous organic solvent in the technological process, compared with prior art more simple, green and safety.
(2) preparation of isoliquiritigenin is that to adopt after polyamide resin separates content be that the liquirtin of 70-90% transforms among the present invention; Therefore isoliquiritigenin content after conversion is higher; Simplified the separation and purification process greatly; Can not only improve the yield of isoliquiritigenin, also guarantee the content (90-98%) of isoliquiritigenin simultaneously.
(3) polymeric amide of the present invention and macroporous resin are reusable, so cost is relatively low, is suitable for large-scale production, can improve the overall availability of Radix Glycyrrhizae greatly, save natural resources of Chinese medicinal materials, produce good social benefit and economic benefit.
In order to understand beneficial effect of the present invention better, below with polymeric amide and further explanation of macroporous resin adsorption experiment conduct.
A. polymeric amide is to the Staticadsorption experiment of liquirtin, Potenlini and carbohydrate content in the different pH licorice extracts
Purpose: the principle of polymeric amide absorption mainly is an adsorption by hydrogen bond, has specificity for the flavones ingredient absorption that contains phenolic hydroxyl group.For the adsorption conditions of flavones in the preferred licorice extract, the polymeric amide absorption of liquirtin, Potenlini and carbohydrate content in the different pH licorice extracts is investigated.
Material: liquorice beverage extracting solution (containing the crude drug amount is 0.2g/mL): extracting liquorice medicinal material 500g, add water 4L and extracted 2 hours, add water 3.5L again and extracted 2 hours, filter, merging filtrate is concentrated into and contains the soup that the crude drug amount is 0.2g/mL.Liquirtin content 1.5mg/mL wherein, glycyrrhizic acid content 6mg/mL, carbohydrate 7.2mg/mL.Polymeric amide (80-100 order, Chemical Reagent Co., Ltd., Sinopharm Group)
Method and result: take by weighing 9 parts of 5g polyamide resins, adding the pH value respectively is 5.0,5.5,6.0; 6.5,7.0,7.5,8.0; 8.5 9.0 liquorice beverage extracting solution 50mL puts in 25 ℃ of water bath chaders, adsorbs 12 hours; Soup after getting the preceding soup of absorption respectively and adsorbing, the HPLC method is measured liquirtin, Potenlini, sulfuric acid anthrone colorimetric method for determining carbohydrate content.Calculate its polymeric amide adsorption rate.
Content * 100% in the soup before adsorption rate (%)=(content in content in the preceding soup of absorption-absorption back soup)/absorption
Table 1 polymeric amide is to the absorption of liquirtin, Potenlini and polyose composition in the different pH licorice extracts
The result shows that different pH are bigger to the Adsorption Effect of liquirtin, when soup pH less than 7.5 the time, because the phenolic hydroxyl group in the liquirtin structure is not ionized, thereby polymeric amide adsorption rate higher (about 90%); And different pH also have remarkably influenced to Potenlini absorption, when the pH value greater than 6.0 the time, all ionizes of the carboxyl in the Potenlini, thereby polymeric amide is not basically to its absorption (less than 5%); Carbohydrate content polarity is very big, and pH is not obvious to its influence, under each pH value, all can not be adsorbed by polymeric amide.
Conclusion: when pH value 6.0-7.5, polymeric amide can be to liquirtin constituents specific adsorption in the Radix Glycyrrhizae soup, and Potenlini and polyose composition are not exerted an influence.
B. the desorption experiment of liquirtin on polymeric amide under the different alcohols concentration
Purpose: investigate the desorption situation of liquirtin on polyamide resin under the different alcohols concentration, preferred suitable eluting solvent.
Material: liquorice beverage extracting solution (containing the crude drug amount is 0.2g/mL), polymeric amide (80-100 order, Chemical Reagent Co., Ltd., Sinopharm Group), ethanol (food grade).
Method and result: take by weighing 6 parts of 5g polyamide resins, add the pH value respectively and be 7.0 liquorice beverage extracting solution 50mL, put in 25 ℃ of water bath chaders; Adsorbed 12 hours, the soup after getting the preceding soup of absorption respectively and adsorbing, the HPLC method is measured the adsorptive capacity (mg) to liquirtin; Filter respectively again,, put in 25 ℃ of water bath chaders adding the 50mL Different concentrations of alcohol aqueous solution in each polyamide resin; Desorption 12 hours is measured the desorption amount (mg) in the soup.Calculate its polymeric amide desorption rate.
Desorption rate (%)=liquirtin desorption amount/liquirtin adsorptive capacity * 100%
The desorption of liquirtin on polymeric amide under the table 2 different alcohols concentration
The result shows that the desorption rate of liquirtin differs greatly under the different alcohols concentration, when determining alcohol greater than 20% the time, can be with the desorption of most of liquirtin.
Conclusion: consider production cost, the eluting solvent of liquirtin constituents on polyamide column is chosen as 20-40% ethanol.
C. different macroporous resins are to the adsorption experiment of Potenlini in the Radix Glycyrrhizae soup and polyose composition
Purpose: investigate the absorption situation of different macroporous resins, preferred suitable macroporous resin to Radix Glycyrrhizae acids and carbohydrate content in the Radix Glycyrrhizae soup.
Material: polymeric amide absorption back liquorice beverage extracting solution (containing the crude drug amount is 0.2g/mL): extracting liquorice aqueous extract (containing the crude drug amount is 0.2g/mL) 1000mL; Regulate pH value to 7.0, filter, again through polymeric amide chromatographic column (polymeric amide dry weight 200g); Collect effluent; Do not contain liquirtin in the effluent, glycyrrhizic acid content 5.68mg/mL, polyose 6.98mg/mL.
Macroporous resin (Cangzhou Bon Adsorption Material Science and Technology Co., Ltd), type is following:
Table 3 macroporous resin type
Method and result: take by weighing all types of macroporous resin 5g; Totally 8 parts, add polymeric amide absorption back liquorice beverage extracting solution 50mL respectively, put in 25 ℃ of water bath chaders; Adsorbed 12 hours; Soup after getting the preceding soup of absorption respectively and adsorbing, the HPLC method is measured Potenlini, sulfuric acid anthrone colorimetric method for determining carbohydrate content.Calculate its adsorption rate.
The different macroporous resins of table 4 are to the absorption of Potenlini and carbohydrate content in the Radix Glycyrrhizae soup
The result shows that carbohydrate content polarity is very big, and all kinds of macroporous resins all do not produce specific adsorption; And different macroporous resins are bigger to the Adsorption Effect of Potenlini, and wherein best with the adsorption effect of HPD400 and AB-8, HPD600, HPD100HPD826, DM130, D101 take second place, and the ADS-7 adsorption rate is the poorest.
Conclusion: preferred HPD400 and AB-8 are as the Potenlini polymeric adsorbent.
D. the desorption experiment of Potenlini on macroporous resin under the different alcohols concentration
Purpose: investigate the desorption situation of Potenlini on HPD400 and AB-8 macroporous resin under the different alcohols concentration, preferred suitable eluting solvent.
Material: polymeric amide absorption back liquorice beverage extracting solution (containing the crude drug amount is 0.2g/mL), D101 and HPD400 macroporous resin (Cangzhou Bon Adsorption Material Science and Technology Co., Ltd), ethanol (food grade).
Method and result: take by weighing 5 parts of macroporous resins (HPD400 and AB-8), add polymeric amide absorption back liquorice beverage extracting solution 50mL respectively, put in 25 ℃ of water bath chaders; Adsorbed 12 hours, the soup after getting the preceding soup of absorption respectively and adsorbing, the HPLC method is measured the adsorptive capacity (mg) to liquirtin; Filter respectively again, in each macroporous resin, add the 50mL Different concentrations of alcohol aqueous solution, put in 25 ℃ of water bath chaders; Desorption 12 hours is measured the desorption amount (mg) in the soup.Calculate its desorption rate.
Desorption rate (%)=Potenlini desorption amount/Potenlini adsorptive capacity * 100%
The desorption of Potenlini on macroporous resin under the table 5 different alcohols concentration
The result shows that the desorption rate of Potenlini differs greatly under the different alcohols concentration, when determining alcohol greater than at 60-80% the time, the desorption best results of Potenlini.
Conclusion: the eluting solvent of Potenlini constituents on macroporous resin column is chosen as 60-80% ethanol.
Description of drawings
Fig. 1 is preparation technology's schema of the present invention
Embodiment
Below through the embodiment form; Foregoing of the present invention is remake further detailed description; But should this be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following instance, all technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Extracting liquorice 5kg, boiling twice adds 8 times of amounts that water is medicinal material weight for the first time; Decoct 2h, add 6 times of amounts that water is medicinal material weight for the second time, decoct 1h; Collecting decoction filters, and filtrating is evaporated to 2.5L (crude drug amount 0.2g/mL) under 0.07MPa, 80 ℃; Add ammoniacal liquor and regulate pH to 6.0, successively through 80-100 order polymeric amide (dry weight is 2.5kg) chromatographic column and AB-8 macroporous resin (dry weight is 2.5kg) chromatographic column;
Licorice polysaccharide preparation: collect effluent, specific density is 1.10 when being concentrated into 65 ℃, adds 95% (v/v) ethanol and makes and contain the alcohol amount and reach 80% (v/v); Stir, left standstill 12 hours, filter; Drying promptly gets licorice polysaccharide 206g, and content is 56.2% (sulfuric acid anthrone colourimetry);
The isoliquiritigenin preparation: the polymeric amide chromatographic column with 40% ethanol elution of 1 times of column volume, is collected ethanol eluate more earlier with 1 times of column volume deionized water wash-out; Decompression recycling ethanol was placed 24 hours, separated out the liquirtin deposition; Filter, get liquirtin bullion 38.3g, content is 78.3% (HPLC method); Get above liquirtin bullion with 40% ethanol 3000mL dissolving, add hydrochloric acid again and make that concentration of hydrochloric acid is 0.5mol/L in the solution, 100 ℃ of acid hydrolysiss 2 hours are put cold; Regulate pH to neutral, continue hydro-oxidation sodium again and make that naoh concentration is 1mol/L in the solution, 80 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 2 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying gets isoliquiritigenin 24.4g again, and content is 95.8% (HPLC method);
The Potenlini preparation: macroporous resin chromatography with 80% ethanol elution of 2 times of column volumes, is collected ethanol eluate more earlier with 1 times of column volume deionized water wash-out; Decompression recycling ethanol, soup is regulated pH to 2, places; Separate out deposition, filter drying; Promptly get Potenlini 169g, content is 77.1% (HPLC method).
Embodiment 2
Extracting liquorice 5kg, refluxing extraction twice adds 8 times of amount 50% ethanol for the first time; Refluxing extraction 2h adds 6 times of amount 50% ethanol for the second time, refluxing extraction 2h; United extraction liquid filters, and filtrating is at 0.12MPa, 60 ℃ of following decompression recycling ethanols; Add water to 2.5L (crude drug amount 0.2g/mL) again, add 10% sodium hydroxide solution and regulate pH to 7.0, successively through 80-100 order polymeric amide (dry weight is 10kg) chromatographic column and AB-8 macroporous resin (dry weight is 10kg) chromatographic column;
Licorice polysaccharide preparation: collect effluent, specific density is 1.15 when being concentrated into 65 ℃, adds 95% (v/v) ethanol and makes and contain the alcohol amount and reach 80% (v/v), stirs, and leaves standstill 24 hours, filters, and drying promptly gets licorice polysaccharide 91g, and content is 63.8% (sulfuric acid anthrone colourimetry);
The isoliquiritigenin preparation: the polymeric amide chromatographic column with 20% ethanol elution of 3 times of column volumes, is collected ethanol eluate more earlier with 3 times of column volume deionized water wash-outs; Decompression recycling ethanol was placed 24 hours, separated out the liquirtin deposition; Filter, get liquirtin bullion 31.3g, content is 87.5% (HPLC method); Get above liquirtin bullion with 40% ethanol 3000mL dissolving, add hydrochloric acid again and make that concentration of hydrochloric acid is 2mol/L in the solution, 90 ℃ of acid hydrolysiss 2 hours are put cold; Regulate pH to neutral, continue hydro-oxidation sodium again and make that naoh concentration is 3mol/L in the solution, 90 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 5 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying gets isoliquiritigenin 21.4g again, and content is 96.2% (HPLC method);
The Potenlini preparation: macroporous resin chromatography with 60% ethanol elution of 6 times of column volumes, is collected ethanol eluate more earlier with 3 times of column volume deionized water wash-outs; Decompression recycling ethanol, soup is regulated pH to 3, places; Separate out deposition, filter drying; Promptly get Potenlini 191g, content is 65.4% (HPLC method).
Embodiment 3
Extracting liquorice 5kg, boiling twice adds 8 times of amounts that water is medicinal material weight for the first time; Decoct 2h, add 6 times of amounts that water is medicinal material weight for the second time, decoct 1h; Collecting decoction filters, and filtrating is evaporated to 5L (crude drug amount 0.1g/mL) under 0.10MPa, 70 ℃; Add ammoniacal liquor and regulate pH to 6.0, successively through 80-100 order polymeric amide (dry weight is 5kg) chromatographic column and AB-8 macroporous resin (dry weight is 5kg) chromatographic column;
Licorice polysaccharide preparation: collect effluent, specific density is 1.15 when being concentrated into 65 ℃, adds 95% (v/v) ethanol and makes and contain the alcohol amount and reach 80% (v/v); Stir, left standstill 36 hours, filter; Drying promptly gets licorice polysaccharide 183g, and content is 78.6% (sulfuric acid anthrone colourimetry);
The isoliquiritigenin preparation: the polymeric amide chromatographic column with 30% ethanol elution of 2 times of column volumes, is collected ethanol eluate more earlier with 2 times of column volume deionized water wash-outs; Decompression recycling ethanol was placed 24 hours, separated out the liquirtin deposition; Filter, get liquirtin bullion 36.4g, content is 83.7% (HPLC method); Get above liquirtin bullion with 40% ethanol 3000mL dissolving, add hydrochloric acid again and make that concentration of hydrochloric acid is 1.0mol/L in the solution, 90 ℃ of acid hydrolysiss 2 hours are put cold; Regulate pH to neutral, continue hydro-oxidation sodium again and make that naoh concentration is 2mol/L in the solution, 90 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 3 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying gets isoliquiritigenin 28.1g again, and content is 98.5% (HPLC method);
The Potenlini preparation: macroporous resin chromatography with 70% ethanol elution of 4 times of column volumes, is collected ethanol eluate more earlier with 2 times of column volume deionized water wash-outs; Decompression recycling ethanol, soup is regulated pH to 2, places; Separate out deposition, filter drying; Promptly get Potenlini 152g, content is 83.4% (HPLC method).
Embodiment 4
Extracting liquorice 5kg, refluxing extraction twice adds 8 times of amount 20% ethanol for the first time; Refluxing extraction 2h adds 6 times of amount 20% ethanol for the second time, refluxing extraction 2h; United extraction liquid filters, and filtrating is at 0.08MPa, 70 ℃ of following decompression recycling ethanols; Add water to 2.5L (crude drug amount 0.2g/mL) again, add ammoniacal liquor and regulate pH to 7.0, successively through 80-100 order polymeric amide (dry weight is 5kg) chromatographic column and AB-8 macroporous resin (dry weight is 5kg) chromatographic column;
Licorice polysaccharide preparation: collect effluent, specific density is 1.20 when being concentrated into 65 ℃, adds 95% (v/v) ethanol and makes and contain the alcohol amount and reach 80% (v/v); Stir, left standstill 24 hours, filter; Drying promptly gets licorice polysaccharide 141g, and content is 61.9% (sulfuric acid anthrone colourimetry);
The isoliquiritigenin preparation: the polymeric amide chromatographic column with 30% ethanol elution of 2 times of column volumes, is collected ethanol eluate more earlier with 2 times of column volume deionized water wash-outs; Decompression recycling ethanol was placed 24 hours, separated out the liquirtin deposition; Filter, get liquirtin bullion 34.9g, content is 80.5% (HPLC method); Get above liquirtin bullion with 40% ethanol 3000mL dissolving, add hydrochloric acid again and make that concentration of hydrochloric acid is 1.5mol/L in the solution, 90 ℃ of acid hydrolysiss 2 hours are put cold; Regulate pH to neutral, continue hydro-oxidation sodium again and make that naoh concentration is 1mol/L in the solution, 90 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 4 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying gets isoliquiritigenin 23.8g again, and content is 96.1% (HPLC method);
The Potenlini preparation: macroporous resin chromatography with 60% ethanol elution of 3 times of column volumes, is collected ethanol eluate more earlier with 3 times of column volume deionized water wash-outs; Decompression recycling ethanol, soup is regulated pH to 3, places; Separate out deposition, filter drying; Promptly get Potenlini 187g, content is 80.6% (HPLC method).
Claims (6)
1. an extraction preparation method who fully utilizes Radix Glycyrrhizae is characterized in that it comprises the steps:
A. extracting liquorice medicinal material through 0～50% extraction using alcohol, reclaim under reduced pressure extracting solution, adds water again and makes in the soup every milliliter to contain crude drug amount 0.1～0.2g, regulates soup pH to 6～7, successively through polymeric amide chromatographic column and macroporous resin chromatography;
B. licorice polysaccharide preparation: collect through the effluent behind the macroporous resin, specific density is 1.10～1.20 when being evaporated to 65 ℃, and adding ethanol to determining alcohol is 80%, places, and separates out deposition, filters, and drying promptly gets licorice polysaccharide;
C. isoliquiritigenin preparation: the polymeric amide chromatographic column with 20～40% ethanol elutions of 1～3 times of column volume, is collected ethanol eluate more earlier with 1～3 times of column volume deionized water wash-out, and decompression recycling ethanol is placed and separated out the liquirtin deposition, filters; The extracting liquorice glycosides is with 40～60% dissolve with ethanol, adds hydrochloric acid again and makes that concentration of hydrochloric acid is 0.5～2mol/L in the solution, and 80～100 ℃ of acid hydrolysiss 2 hours are put cold; Hydro-oxidation sodium is regulated pH to neutral, continues hydro-oxidation sodium again and makes that naoh concentration is 1～3mol/L in the solution, and 80～100 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 2～5 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying promptly gets isoliquiritigenin again;
D. Potenlini preparation: macroporous resin chromatography with 60～80% ethanol elutions of 2～6 times of column volumes, is collected ethanol eluate more earlier with 1～3 times of column volume deionized water wash-out; Decompression recycling ethanol, glycyrrhizic aqueous solution is regulated pH to 2～3 with hydrochloric acid, places; Separate out deposition; Filter, drying promptly gets Potenlini.
2. the extraction preparation method of comprehensive utilization according to claim 1 Radix Glycyrrhizae is characterized in that the specification of used polymeric amide is the 80-100 order, with the weight ratio of licorice medicinal materials be 2: 1～4; The model of big pore resin is AB-8 or HPD-400, with the weight ratio of licorice medicinal materials be 2: 1～4.
3. the extraction preparation method of comprehensive utilization Radix Glycyrrhizae according to claim 1; It is characterized in that the isoliquiritigenin preparation method is: the polymeric amide chromatographic column is earlier with 2 times of column volume deionized water wash-outs, again with 30% ethanol elution of 2 times of column volumes; Collect ethanol eluate; Decompression recycling ethanol is placed and is separated out the liquirtin deposition, filters; The extracting liquorice glycosides is with 50% dissolve with ethanol, adds hydrochloric acid again and makes that concentration of hydrochloric acid is 1mol/L in the solution, and 90 ℃ of acid hydrolysiss 2 hours are put cold; Regulate pH to neutral, continue hydro-oxidation sodium again and make that naoh concentration is 2mol/L in the solution, 90 ℃ of heating alkali open rings 2 hours are put cold; Regulate pH to 3 with hydrochloric acid, place, separate out crystallization, filter; With 50% ethyl alcohol recrystallization, drying promptly gets isoliquiritigenin again.
4. the extraction preparation method of comprehensive utilization Radix Glycyrrhizae according to claim 1 is characterized in that the preparation method of Potenlini is: macroporous resin chromatography is earlier with 2 times of column volume deionized water wash-outs; With 70% ethanol elution of 4 times of column volumes, collect ethanol eluate, decompression recycling ethanol again; Soup is regulated pH to 2, places, and separates out deposition; Filter, drying promptly gets Potenlini.
5. the extraction preparation method of comprehensive utilization Radix Glycyrrhizae according to claim 1, it is characterized in that the condition of said decompression recycling ethanol is: pressure is 0.07～0.12MPa, temperature is 60～80 ℃.
6. the extraction preparation method of comprehensive utilization Radix Glycyrrhizae according to claim 2 is characterized in that the weight ratio of polymeric amide and licorice medicinal materials is 1: 1; The weight ratio of macroporous resin and licorice medicinal materials is 1: 1.
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| CN103599172A (en) * | 2013-10-09 | 2014-02-26 | （株）Ad生物技术 | Method of preparing liquorice extract |
| CN103833806A (en) * | 2012-11-22 | 2014-06-04 | 天津药物研究院 | Preparation method of traditional Chinese medicine chemical component |
| CN104861015A (en) * | 2015-01-16 | 2015-08-26 | 李玉山 | Synergistic clean extraction method of effective components in licorice root |
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| CN105031178A (en) * | 2015-08-03 | 2015-11-11 | 南京中医药大学 | Extracting refining method making efficient utilization of anemarrhena asphodeloides |
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| CN105669543A (en) * | 2015-12-31 | 2016-06-15 | 中国药科大学 | Isoliquiritigenin nicotinamide eutectic crystal and preparation method thereof |
| CN105777521A (en) * | 2016-05-11 | 2016-07-20 | 西安兴博凯生物科技有限责任公司 | Industrial separating and purifying method for licoflavone series monomer |
| CN110090469A (en) * | 2019-04-26 | 2019-08-06 | 青岛农业大学 | A method of extraction purification glycyrrhizin and enoxolone from Radix Glycyrrhizae |
| CN113666977A (en) * | 2021-08-17 | 2021-11-19 | 甘肃亚兰药业有限公司 | Production process for combined separation of multiple active ingredients of liquorice |
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