CN103860631A - Method for preparing liquorice active substances - Google Patents

Method for preparing liquorice active substances Download PDF

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CN103860631A
CN103860631A CN201410075183.3A CN201410075183A CN103860631A CN 103860631 A CN103860631 A CN 103860631A CN 201410075183 A CN201410075183 A CN 201410075183A CN 103860631 A CN103860631 A CN 103860631A
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ethanol
precipitate
supernatant
concentrated
obtains
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高彦祥
樊蕊
李铁军
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BAOTOU HUIXIN INDUSTRIAL Co Ltd
INNER MONGOLIA PUFANSHENG IOLOGICAL SCIENCE & TECHNOLOGY Co Ltd
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BAOTOU HUIXIN INDUSTRIAL Co Ltd
INNER MONGOLIA PUFANSHENG IOLOGICAL SCIENCE & TECHNOLOGY Co Ltd
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Abstract

The invention discloses a method for preparing liquorice active substances. The method comprises the following steps: pretreatment and hot water extraction treatment of raw materials and ultrasonic auxiliary extraction treatment, alcohol precipitation treatment, purification treatment, acid precipitation treatment, redissolution treatment, ammoniated treatment, acidification treatment, crystallization and purification of liquorice residues. The method can be used for fully utilizing liquorice source, and achieving the separation of different effective components during extraction to avoid the loss of other several active substances during single extraction; the production cost is low, the operation is simple, the production efficiency is high, and no environment pollution is produced; by utilizing the method, continuous production is achieved, and the products obtained according to the method have high quality; the method is easily popularized and utilized and provides a new approach for comprehensive utilization of liquorice; by utilizing the method, the comprehensive utilization value of liquorice is improved.

Description

A kind of preparation method of Radix Glycyrrhizae active substance
Technical field
The present invention relates to a kind of Chinese herbal medicine comprehensive utilization technique, specifically, relate to a kind of preparation method of Radix Glycyrrhizae active substance.
Background technology
Radix Glycyrrhizae (Glycyrrhizauralensis) another name Herba Hedyotis cantonensis root, Radix Glycyrrhizae, state are old, Herba Hedyotis cantonensis, sweet root etc., are one of conventional Chinese herbal medicine of China's tradition, are pulse family (Leguminosae) Glycyrrhiza (GlycyrrhizaLinn) herbaceos perennial.The flat sweet in the mouth of Radix Glycyrrhizae property, has effect of invigorating the spleen and replenishing QI, heat-clearing and toxic substances removing, expelling phlegm for arresting cough, relieving spasm to stop pain, coordinating the actions of various ingredients in a prescription, has extremely wide clinical value.In China, Radix Glycyrrhizae is the medical herbs simply of consumption maximum in 2000 plurality of herbal, have saying of " ten side's nine grass, without not Cheng Fang of grass ".Meanwhile, best a kind of natural sweetener that Radix Glycyrrhizae is still found up to now.Radix Glycyrrhizae is distributed widely between north latitude 30-55 °, and arid, the semi-arid areas of 40 ° of left and right of north latitude is referred to as " Radix Glycyrrhizae zonation ", the main place of production in the Central Asia, North America and Eastern Europe, especially take the Central Asia and Mediterranean be distribution center.Radix Glycyrrhizae is concentrated and is distributed in three northern areas of China (the each provinces and regions in northeast, North China and northwest), producing region centered by Xinjiang, the Inner Mongol, Ningxia and Gansu in China.
The composition of medicinal Radix Glycyrrhizae quality and its chemical composition, cumulative change have direct relation.Successively from glycyrrhiza genus, extract, separate, identified 200 number of chemical compositions, wherein most important and to have confirmed to have bioactive composition be mainly glycyrrhizic acid, triterpene saponin, flavonoid, Coumarins, polysaccharide, alkaloid, aminoacid etc.Pharmacological research shows, glycyrrhizic acid and enoxolone have removing toxic substances, antiinflammatory, analgesia, antineoplastic action.In recent years, Radix Glycyrrhizae is also for preventing and treating viral hepatitis, cancer and acquired immune deficiency syndrome (AIDS) etc.Glycyrrhizin has the integrated feature of sugared medicine.Except medical value, Radix Glycyrrhizae has important purposes in the industry such as food, beverage, Medicated cigarette, cosmetics.
China is Radix Glycyrrhizae big producing country, exports more than 30 countries and regions, and only Japanese annual requirement just reaches ten thousand tons more than of l.But the further investigation of biosynthesis, mechanism of action, refining processing and comprehensive utilization to the main effective ingredient glycyrrhizic acid of Radix Glycyrrhizae also lags behind the states such as Japan.Be subject to technical conditions and process technology limit, the purity of the glycyrrhizic acid inclusion compound of application traditional method for extracting is low, and price is in the international market low, and extraction efficiency is also low, insufficient to utilizing of Licorice, has affected the development of China's Radix Glycyrrhizae industry.
At present, licorice root products mostly is the primary extract of glycyrrhizic acid, licoflavone, polysaccharide, and little for the single step processing of Radix Glycyrrhizae, does not prepare the method for monoammonium glycyrrhizinate, licoflavone and Angelica Polysaccharide in existing documents and materials about single step.Chinese patent (CN102836202A) discloses the method for Radix Glycyrrhizae aerial parts comprehensive development and utilization, utilizes organic reagent to extract licorice extract, is obtaining crude flavonoid powder through concentrating.This patent document does not relate to the preparation of monoammonium glycyrrhizinate, and this method has the possibility of certain dissolvent residual, has potential safety problem.Chinese patent (CN101766680A) has been announced comprehensive extraction method of glycyrrhiza, extracts with different solvents through 3 times, obtains thick glycyrrhizic acid, Angelica Polysaccharide and licoflavone, and this step complicated and time consumption and product purity are low.
Summary of the invention
Technical problem solved by the invention is to provide a kind of preparation method of Radix Glycyrrhizae active substance, has realized deep processing and comprehensive utilization to Radix Glycyrrhizae, has expanded its range of application, has improved its added value.
Technical scheme is as follows:
A preparation method for Radix Glycyrrhizae active substance, comprising:
The pretreatment of raw material; Radix Glycyrrhizae is screened, removed various impurity, wash, be dried, pulverized 60 mesh sieves;
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I, and extraction temperature is 75-95 ℃, and solid-liquid ratio is 1:5-1:15, and extracting times is 1-3 time, and each extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue obtaining is for ultrasonic Assisted Extraction, obtain lixiviating solution II, ultrasonic power 300-500W, extraction temperature is 60-80 ℃, solid-liquid ratio is 1:10-1:20, and extracting times is 1-2 time, and each extraction time is 20-30min, lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing, coarse filtration liquid I is through centrifugal treating, rotating speed 4200r/min, centrifugal 10min, obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, being concentrated into soluble solid is 30-40Brix, obtain concentrated solution I, concentrated solution I adds 75-95% ethanol to make it ethanol final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 25-35Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75-95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; Final precipitate with ethanol precipitate I is used under room temperature to 60-90% washing with alcohol 1-3 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, and condenser temperature is-40 ℃~-80 ℃, and be 36-48h drying time, obtains Angelica Polysaccharide.
Further: also comprise the heavy step of processing, redissolving processing, ammonification processing, acidification and crystallization of acid, wherein,
Acid is heavy to be processed; Adopt vacuum evaporator concentrated to remove ethanol final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, vacuum is 0.08MPa, is concentrated into soluble solid for being concentrated into 30-35Brix, makes concentration of alcohol lower than 5%, obtain concentrated solution II, concentrated solution II is diluted and made soluble solid reach 15Brix with deionized water, use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 1.5-2.5, obtain the heavy liquid I of acid;
Redissolve and process; Heavy acid liquid I, in 4 ℃ of standing 12h, with rotating speed 4200r/min, is carried out to centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent I, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 30-60min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 7.5-9.0 with strong aqua ammonia, stir 1h, static layering, obtains phase I, lower phase I;
Acidification; Upper phase I is used below glacial acetic acid adjust pH to 4.0,70-90 ℃ of water bath heat preservation 30min, stand at low temperature 6-12h, obtains acidifying solution I;
Crystallization; Acidifying solution I adds dehydrated alcohol dilution, make concentration of alcohol reach 90%, stand at low temperature 4-8h, with the centrifugal 20min of 4200r/min, obtain supernatant II and precipitation after centrifugal, the precipitation after centrifugal is carried out vacuum lyophilization, vacuum is 0.014MPa, condenser temperature is-40 ℃~-60 ℃, and be 20-30h drying time, after precipitation is dry, obtains monoammonium glycyrrhizinate.
Further: also to comprise purification step, supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, being concentrated into dry doubling 50% ethanol redissolves, utilize macroporous resin to carry out purification, resin is selected XDA-8, LX20B, D101, AB-8 is wherein a kind of, and the 8-12h that vibrates under room temperature carries out static adsorption, the 8-12h that vibrates under 70-90% ethanol room temperature carries out desorbing, collection eluent carries out vacuum evaporator and concentrates to remove ethanol, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 30-35Brix, sprays dry.The dry inlet temperature 160-200 ℃ of spraying, outlet temperature 80-85 ℃, obtains licoflavone after being dried.
Further: hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 75 ℃, and solid-liquid ratio is 1:5, and extracting times is 1 time, and extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained for ultrasonic Assisted Extraction, obtains lixiviating solution II, ultrasonic Assisted Extraction condition: power 300W, extraction temperature are that 60 ℃, solid-liquid ratio are that 1:10, extracting times are that 1 time, extraction time are 20min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing, coarse filtration liquid I process centrifugal treating, rotating speed 4200r/min, centrifugal 10min, obtains clear liquor I, and clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 30Brix, obtain concentrated solution I, concentrated solution I adds 75% ethanol to make it ethanol final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 25Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 60% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%, obtains concentrated solution II, concentrated solution II is diluted and made soluble solid reach 15Brix with deionized water, use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 1.5, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent ethyl acetate, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 30min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 7.5 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 4.0, guarantee that acetic acid is excessive, 70 ℃ of water bath heat preservation 30min, stand at low temperature 6h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 4h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-40 ℃, and be 20h drying time, obtains monoammonium glycyrrhizinate after drying precipitate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, and are concentrated into dry doubling 50% ethanol and redissolve.Utilize XDA-8 macroporous resin to carry out purification, the 8h that vibrates under room temperature carries out static adsorption suction, the 8h that vibrates under eluent 70% ethanol room temperature carries out desorbing, it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry; 160 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
Further: hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I, and extraction temperature is 85 ℃, and solid-liquid ratio is 1:15, and extracting times is 3 times, and each extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition: power 500W, extraction temperature are that 70 ℃, solid-liquid ratio are that 1:10, extracting times are that 2 times, extraction time are 30min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing; Coarse filtration liquid I process centrifugal treating, with rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 85% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 70% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%; The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix; Use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 2.0, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent ethyl acetate, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 40min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 8.0 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.5,80 ℃ of water bath heat preservation 30min, stand at low temperature 8h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 6h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-50 ℃, and be 25h drying time; After drying precipitate, obtain monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, be concentrated into dry doubling 50% ethanol and redissolve, utilize macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, the 10h that vibrates under eluent 70% ethanol room temperature carries out desorbing, collection eluent carries out vacuum evaporator and concentrates to remove ethanol, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 30Brix, sprays dry.180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
Further: hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I, and extraction temperature is 85 ℃, and solid-liquid ratio is 1:10, and extracting times is 2 times, and each extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained, for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition, power 400W, extraction temperature is 70 ℃, and solid-liquid ratio is 1:20, and extracting times is 1 time, and extraction time is 30min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing; Coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 85% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 85% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 70% washing with alcohol 2 times, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%, and the concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix; Use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 2.5, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent dichloromethane, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 40min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 9.0 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.5,80 ℃ of water bath heat preservation 30min, stand at low temperature 8h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 6h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-50 ℃, and be 25h drying time, after precipitation is dry, obtains monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, vacuum concentration to dry doubling redissolves with 50% ethanol, utilize macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, and the 10h that vibrates under eluent 80% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry; 180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
Further: hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 85 ℃, and solid-liquid ratio is 1:15, and extracting times is 2 times, and each extraction time is 3h; Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition: power 400W, extraction temperature are that 80 ℃, solid-liquid ratio are that 1:15, extracting times are that 2 times, extraction time are 30min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing, coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min, obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 95% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitation is redissolved with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 80% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-60 ℃, be that 48h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Be concentrated into soluble solid to 35Brix, make concentration of alcohol lower than 5%, the concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix; Use volume ratio 1:1 aqueous sulfuric acid to stir and add, adjust pH to 2.0, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved with organic solvent water saturation n-butyl alcohol, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 60min, make it to dissolve completely, obtain organic solution I;
Ammonification processing; Organic liquor I is adjusted to pH to 8.5 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.0,80 ℃ of water bath heat preservation 30min, stand at low temperature 10h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 8h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-60 ℃, and be 30h drying time; After precipitation is dry, obtain monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, be concentrated into dry doubling 50% ethanol and redissolve, utilize macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, the 10h that vibrates under eluent 80% ethanol room temperature carries out desorbing, collection eluent carries out vacuum evaporator and concentrates to remove ethanol, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, sprays dry; 180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures, obtain licoflavone after being dried.
Further: hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I; Extraction temperature is 95 ℃, and solid-liquid ratio is 1:10, and extracting times is 3 times, and each extraction time is 3h; Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained, for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition, power 500W, extraction temperature is 80 ℃, and solid-liquid ratio is 1:15, and extracting times is 2 times, and extraction time is 30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing; Coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 40Brix, obtain concentrated solution I, concentrated solution I adds 95% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 35Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 90% washing with alcohol 3 times, collecting precipitation, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-80 ℃, be that 48h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Be concentrated into soluble solid 30Brix, make concentration of alcohol lower than 5%; The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix.Use volume ratio 1:1 aqueous sulfuric acid to stir and add, adjust diluent pH value to 1.5, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved with organic solvent water saturation n-butyl alcohol, according to solid-to-liquid ratio 1:3, stirring and dissolving precipitation I 50min, makes it to dissolve completely, obtains organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 8.5 with strong aqua ammonia, magnetic agitation 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.0,90 ℃ of water bath heat preservation 30min, stand at low temperature 12h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 8h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-60 ℃, and be 30h drying time; After drying precipitate, obtain monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, being concentrated into dry doubling 50% ethanol redissolves, utilize macroporous resin to carry out purification, the 12h that vibrates under room temperature carries out static adsorption suction, and the 12h that vibrates under eluent 90% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry; 200 ℃ of the dry inlet temperatures of spraying, 85 ℃ of outlet temperatures; After dry, obtain licoflavone.
Compared with prior art, the technology of the present invention effect comprises:
1, the present invention takes full advantage of Licorice, has realized the separation of different active components in the process of extracting, the loss of other several active substances while avoiding single extraction.And production cost is lower, it is simple and easy to operate, production efficiency is high, environmentally safe, can realizes serialization and produce, and the product quality obtaining is good, be easy to promote and utilize, for the comprehensive utilization of Radix Glycyrrhizae provides a new way, improved the comprehensive utilization value of Radix Glycyrrhizae.Radix Glycyrrhizae active substance can be used as functional factor, is applied to food, medicine processing, has realized deep processing and comprehensive utilization to Radix Glycyrrhizae, has expanded its range of application, has improved its added value.
2, social benefit and economic benefit:
(1) the main range of application of the present invention and market orientation: Radix Glycyrrhizae active substance can be used as functional factor, is applied to food, medicine processing, has realized deep processing and comprehensive utilization to Radix Glycyrrhizae, has expanded its range of application, has improved its added value.Licoflavone, the Angelica Polysaccharide that the present invention produces can be used as quality raw materials or the adjuvant of some functional food exploitations, be applied to food, medicine processing, be added in various food, health product and pharmaceutical products, main purpose is the physiological action such as antioxidation, raising immunity.Monoammonium glycyrrhizinate is natural sweetener, highly effective and safe, and be a kind of functional factor, hepatoprotective, removing toxic substances.So one aspect of the present invention increases product health care, improves on the other hand product quality and added value etc.
(2) application benefit of the present invention: considerable economic benefit and good social benefit.The present invention is directly benefited for enterprises such as the elementary processing of Radix Glycyrrhizae, carries out the deep processing of Radix Glycyrrhizae, provides reliable approach to the comprehensive utilization of Radix Glycyrrhizae.
(3) promotional value: Radix Glycyrrhizae is the vegetable drug in short supply in China and even the whole world always, Radix Glycyrrhizae has been realized artificial culture at present, and the present invention, take Artificial Cultivation Glycyrrhiza uralensis as raw material, produces active substance, to greatest extent Radix Glycyrrhizae is carried out to comprehensive exploitation, improved its value.
Accompanying drawing explanation
Fig. 1 is the process chart of a kind of preparation method of Radix Glycyrrhizae active substance in the present invention;
Fig. 2 is the canonical plotting (with glucose meter) of Angelica Polysaccharide in the present invention;
Fig. 3 a is the canonical plotting of glycyrrhizic acid in the present invention;
Fig. 3 b is the high-efficient liquid phase analysis result figure of monoammonium glycyrrhizinate standard substance in the present invention;
Fig. 3 c is the high-efficient liquid phase analysis result figure of monoammonium glycyrrhizinate product salt in the present invention;
Fig. 4 a is the UV scanning figure of crude flavonoid powder in the present invention;
Fig. 4 b is the canonical plotting (in rutin) of flavone in the present invention.
The specific embodiment
The present invention, take Radix Glycyrrhizae as raw material, obtains Radix Glycyrrhizae active substance (licoflavone, monoammonium glycyrrhizinate, Angelica Polysaccharide) by substep lixiviate, separation and purification, drying and other steps.
Below with reference to accompanying drawing and preferred embodiment, technical solution of the present invention is elaborated.
As shown in Figure 1, be the process chart of a kind of preparation method of Radix Glycyrrhizae active substance in the present invention.
A preparation method for Radix Glycyrrhizae active substance, step comprises:
Step 1: the pretreatment of raw material;
Radix Glycyrrhizae is screened, removed various impurity, then washed, remove dust, be dry, pulverize 60 mesh sieves.
Step 2: hot water lixiviate processing;
Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 75-95 ℃, and solid-liquid ratio is 1:5-1:15 (mass volume ratio w/v), and extracting times is 1-3 time, and each extraction time is 3h.Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use.
Step 3: the ultrasonic Assisted Extraction processing of glycyrrhiza residue;
The filtering residue being obtained by step 2, for ultrasonic Assisted Extraction, obtains lixiviating solution II.Ultrasonic power 300-500W, extraction temperature is 60-80 ℃, solid-liquid ratio is 1:10-1:20(mass volume ratio w/v), extracting times is 1-2 time, each extraction time is 20-30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards.
Step 4: precipitate with ethanol processing;
Coarse filtration liquid I is through centrifugal treating, rotating speed 4200r/min, centrifugal 10min, obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, being concentrated into soluble solid is 30-40Brix, obtain concentrated solution I, concentrated solution I adds 75-95% ethanol to make it ethanol final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(mass volume ratio w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 25-35Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75-95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5(mass volume ratio/v), 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I.
Step 5: purification process;
Final precipitate with ethanol precipitate I is used under room temperature to 60-90% washing with alcohol 1-3 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, and condenser temperature is-40 ℃~-80 ℃, and be 36-48h drying time, obtains Angelica Polysaccharide.
Step 6: acid is heavy to be processed;
Adopt vacuum evaporator concentrated to remove ethanol final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, vacuum is 0.08MPa, is concentrated into soluble solid for being concentrated into 30-35Brix, makes concentration of alcohol lower than 5%, obtain concentrated solution II, concentrated solution II is diluted and is made soluble solid reach 15Brix with deionized water, use 1:1(volume ratio v/v) aqueous sulfuric acid, stirs and adds, adjust diluent pH value to 1.5-2.5, obtain the heavy liquid I of acid.
Step 7: redissolve and process;
Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, precipitates organic solvent I (one in ethyl acetate, water-saturated n-butanol, dichloromethane) for I and redissolves, according to solid-to-liquid ratio 1:3(mass volume ratio w/v) stirring and dissolving precipitation I 30-60min, make it to dissolve completely, obtain organic liquor I.
Step 8: ammonification processing;
Organic liquor I is adjusted to pH to 7.5-9.0 with strong aqua ammonia, stir 1h, static layering, obtains phase I, lower phase I.
Step 9: acidification;
Upper phase I (water) is used to glacial acetic acid adjust pH to 4.0 following (guaranteeing that acetic acid is excessive), 70-90 ℃ of water bath heat preservation 30min, stand at low temperature 6-12h, obtains acidifying solution I.
Step 10: crystallization;
Acidifying solution I adds dehydrated alcohol dilution, make concentration of alcohol reach 90%, stand at low temperature 4-8h, with the centrifugal 20min of 4200r/min, obtain supernatant II and precipitation II after centrifugal, the precipitation II after centrifugal is carried out vacuum lyophilization, vacuum is 0.014MPa, condenser temperature is-40 ℃~-60 ℃, and be 20-30h drying time, after precipitation is dry, obtains monoammonium glycyrrhizinate.
Step 11: purification.
Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, being concentrated into dry doubling 50% ethanol redissolves, (resin is selected XDA-8 to utilize macroporous resin to carry out purification, XDB-8, LX20B, D101, AB-8 is wherein a kind of), the 8-12h that vibrates under room temperature carries out static adsorption, and the 8-12h that vibrates under 70-90% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30-35Brix, sprays dry.The dry inlet temperature 160-200 ℃ of spraying, outlet temperature 80-85 ℃, obtains licoflavone after being dried.
Product Validation:
1) checking of Angelica Polysaccharide
The checking of Angelica Polysaccharide and content assaying method adopt phenolsulfuric acid colorimetry (with glucose meter).
As shown in Figure 2, be the canonical plotting of Angelica Polysaccharide in the present invention, wherein, abscissa is glucose quality, and vertical coordinate is light absorption value, and by the drafting of standard curve, matching obtains normal equation: y=0.012x-0.006, R 2=0.999, in formula: Y represents absorbance, X represents that (g), R represents correlation coefficient to μ to glucose quality.By normal equation, calculate the content of glucose according to measured light absorption value in experiment, thereby obtain the content (with glucose meter) of polysaccharide.
2) Radix Glycyrrhizae crude flavonoid powder assay method
Radix Glycyrrhizae crude flavonoid powder is suitably diluted with percent by volume 50% ethanol-water solution, and in the analysis of ultraviolet-spectrophotometer, ultraviolet all band scanning result is as Fig. 4 a.Abscissa is scanning wavelength, and vertical coordinate is light absorption value, the demonstration of Fig. 4 a result, and respectively there is an absorption band in Radix Glycyrrhizae crude flavonoid powder, illustrates that it is flavone compound between 300~400nm and between 240~280nm, the method is the qualitative method of flavone.UV scanning can illustrate that this material has the characteristic absorption peak of flavone compound, illustrates that this material is flavone compound, is the qualitative method of flavone.
The assay method of Radix Glycyrrhizae crude flavonoid powder content uses sodium nitrite-sodium hydroxide colorimetry (in rutin), and gained standard curve is shown in Fig. 4 b.Abscissa is rutin concentration, and vertical coordinate is light absorption value.By the drafting of standard curve, matching obtains its standard curve equation: y=0.516x+0.025, R 2=0.999, in formula: Y represents absorbance, X represents rutin concentration (μ g/mL), and R represents correlation coefficient.By normal equation, calculate the content of rutin according to measured light absorption value in experiment, thereby obtain the content (in rutin) of flavone.
3) monoammonium glycyrrhizinate
Shown in Fig. 3 a, it is the canonical plotting of glycyrrhizic acid in the present invention; In the present invention, assay method for glycyrrhizic acid adopts high-efficient liquid phase technique, and wherein abscissa is the concentration of glycyrrhizic acid standard substance, and vertical coordinate is peak area, by the drafting of standard curve, obtains standard curve equation: y=5.926x+11.21, R by matching 2=0.999, in formula: Y represents peak area, X represents the concentration (μ g/mL) of glycyrrhizic acid standard substance, and R represents correlation coefficient.By normal equation, calculate the content of glycyrrhizic acid according to measured peak area in experiment, thereby obtain the content (in glycyrrhizic acid) of monoammonium glycyrrhizinate.
Shown in Fig. 3 b, be the high-efficient liquid phase analysis result figure of monoammonium glycyrrhizinate standard substance in the present invention; As shown in Figure 3 c, be the high-efficient liquid phase analysis result figure of monoammonium glycyrrhizinate product salt in the present invention, abscissa is the time, doing coordinate is peak area.The appearance time of the surveyed material that is designated as on figure, be respectively 6.04 and 5.95min, by the high-efficient liquid phase analysis result of monoammonium glycyrrhizinate product salt is compared and is therefore thought for same material, that is: monoammonium glycyrrhizinate with the high-efficient liquid phase analysis result of glycyrrhizic acid standard substance.
Embodiment 1
A preparation method for Radix Glycyrrhizae active substance, its production stage is:
(1) pretreatment of raw material: Radix Glycyrrhizae is screened, removed various impurity, then washed, remove dust, dry, shattered 60 mesh sieves.
(2) hot water lixiviate processing: Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 75 ℃, and solid-liquid ratio is 1:5(w/v), extracting times is 1 time, extraction time is 3h.Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use.
(3) the ultrasonic Assisted Extraction processing of glycyrrhiza residue: the filtering residue of step (2) gained, for ultrasonic Assisted Extraction, obtains lixiviating solution II, ultrasonic Assisted Extraction condition: power 300W, extraction temperature is 60 ℃, solid-liquid ratio is 1:10(w/v), extracting times is 1 time, extraction time is 20min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards.
(4) precipitate with ethanol processing: coarse filtration liquid I process centrifugal treating, rotating speed 4200r/min, centrifugal 10min, obtains clear liquor I, and clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 30Brix, obtain concentrated solution I, concentrated solution I adds 75% ethanol to make it ethanol final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 25Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I.
(5) purification process: by final precipitate with ethanol precipitate I under room temperature by 60% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time.
(6) acid is heavy processes: adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%, obtains concentrated solution II, concentrated solution II is diluted and is made soluble solid reach 15Brix with deionized water, use 1:1(v/v) aqueous sulfuric acid, stirs and adds, adjust diluent pH value to 1.5, obtain the heavy liquid I of acid.
(7) redissolve and process: acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent ethyl acetate, according to solid-to-liquid ratio 1:3(w/v), stirring and dissolving precipitation I 30min, makes it to dissolve completely, obtains organic liquor I.
(8) ammonification processing: organic solution I is adjusted to pH to 7.5 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I.
(9) acidification: use glacial acetic acid adjust pH to guarantee that to 4.0(acetic acid is excessive upper phase I), 70 ℃ of water bath heat preservation 30min, stand at low temperature 6h, obtains acidifying solution I.
(10) crystallization treatment: acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, stand at low temperature 4h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II.Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-40 ℃, and be 20h drying time, obtains monoammonium glycyrrhizinate after drying precipitate.
(11) purification process: supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, and are concentrated into dry doubling 50% ethanol and redissolve.Utilize XDA-8 macroporous resin to carry out purification, the 8h that vibrates under room temperature carries out static adsorption suction, the 8h that vibrates under eluent 70% ethanol room temperature carries out desorbing, it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, and vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix,, spray dry.160 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
Monoammonium glycyrrhizinate yield 1.23% prepared by this condition, purity is 58%; Licoflavone yield 2.23%, purity is 30%; Angelica Polysaccharide yield 11.27%, purity is 57.8%.
Embodiment 2
A preparation method for Radix Glycyrrhizae active substance, its production stage is:
(1) pretreatment of raw material: Radix Glycyrrhizae is screened, removed various impurity, then washed, remove dust, dry, shattered 60 mesh sieves.
(2) hot water lixiviate processing: Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 85 ℃, and solid-liquid ratio is 1:15 (w/v), and extracting times is 3 times, and each extraction time is 3h.Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use.
(3) the ultrasonic Assisted Extraction processing of glycyrrhiza residue: the filtering residue of step (2) gained is used for to ultrasonic Assisted Extraction, obtain lixiviating solution II, ultrasonic Assisted Extraction condition: power 500W, extraction temperature is 70 ℃, solid-liquid ratio is 1:10 (w/v), extracting times is 2 times, and extraction time is 30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards.
(4) precipitate with ethanol processing: coarse filtration liquid I process centrifugal treating, with rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 85% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I.
(5) purification process: by final precipitate with ethanol precipitate I under room temperature by 70% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time.
(6) acid is heavy processes: adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%.The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix.Use 1:1(v/v) aqueous sulfuric acid, stir and add, adjust diluent pH value to 2.0, obtain the heavy liquid I of acid.
(7) redissolve and process: acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent ethyl acetate, according to solid-to-liquid ratio 1:3(w/v), stirring and dissolving precipitation I 40min, makes it to dissolve completely, obtains organic liquor I.
(8) ammonification processing: organic solution I is adjusted to pH to 8.0 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I.
(9) acidification: use glacial acetic acid adjust pH to guarantee that to 3.5(acetic acid is excessive upper phase I), 80 ℃ of water bath heat preservation 30min, stand at low temperature 8h, obtains acidifying solution I.
(10) crystallization treatment: acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, stand at low temperature 6h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II.Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-50 ℃, and be 25h drying time; After drying precipitate, obtain monoammonium glycyrrhizinate.
(11) purification process: supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, and are concentrated into dry doubling 50% ethanol and redissolve, and utilize LX20B macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, the 10h that vibrates under eluent 70% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, and thickening temperature is 45 ℃, vacuum is 0.08MPa, being concentrated into soluble solid is 30Brix,, spray dry.180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
Monoammonium glycyrrhizinate yield 1.61% prepared by this condition, purity is 60.2%; Licoflavone yield 2.92%, purity is 31.8%; Angelica Polysaccharide yield 9.66%, purity is 61.3%.
Embodiment 3
A preparation method for Radix Glycyrrhizae active substance, its production stage is:
(1) pretreatment of raw material: Radix Glycyrrhizae is screened, removed various impurity, then washed, remove dust, dry, shattered 60 mesh sieves.
(2) hot water lixiviate processing: Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 85 ℃, and solid-liquid ratio is 1:10 (w/v), and extracting times is 2 times, and each extraction time is 3h.Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use.
(3) the ultrasonic Assisted Extraction processing of glycyrrhiza residue: the filtering residue of step (2) gained is used for to ultrasonic Assisted Extraction, obtain lixiviating solution II, ultrasonic Assisted Extraction condition, power 400W, extraction temperature is 70 ℃, solid-liquid ratio is 1:20 (w/v), and extracting times is 1 time, and extraction time is 30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards.
(4) precipitate with ethanol processing: coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 85% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 85% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I.
(5) purification process: by final precipitate with ethanol precipitate I under room temperature by 70% washing with alcohol 2 times, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time.
(6) acid is heavy processes: adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%.The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix.Use 1:1(volume ratio v/v) aqueous sulfuric acid, stir and add, adjust diluent pH value to 2.5, obtain the heavy liquid I of acid.
(7) redissolve and process: acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent dichloromethane, according to solid-to-liquid ratio 1:3(w/v), stirring and dissolving precipitation I 40min, makes it to dissolve completely, obtains organic liquor I.
(8) ammonification processing: organic solution I is adjusted to pH to 9.0 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I.
(9) acidification: use glacial acetic acid adjust pH to guarantee that to 3.5(acetic acid is excessive upper phase I), 80 ℃ of water bath heat preservation 30min, stand at low temperature 8h, obtains acidifying solution I.
(10) crystallization treatment: acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, stand at low temperature 6h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II.Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-50 ℃, and be 25h drying time, after precipitation is dry, obtains monoammonium glycyrrhizinate.
(11) purification process: supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, vacuum concentration to dry doubling redissolves with 50% ethanol, utilize D101 macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, and the 10h that vibrates under eluent 80% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry.180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
Monoammonium glycyrrhizinate yield 1.45% prepared by this condition, purity is 59.7%; Licoflavone yield 2.52%, purity is 28.8%; Angelica Polysaccharide yield 11.32%, purity is 61.3%.
Embodiment 4
A preparation method for Radix Glycyrrhizae active substance, its production stage is:
(1) pretreatment of raw material: Radix Glycyrrhizae is screened, removed various impurity, then washed, remove dust, dry, shattered 60 mesh sieves.
(2) hot water lixiviate processing: Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 85 ℃, and solid-liquid ratio is 1:15 (w/v), and extracting times is 2 times, and each extraction time is 3h.Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use.
(3) the ultrasonic Assisted Extraction processing of glycyrrhiza residue: the filtering residue of step (2) gained is used for to ultrasonic Assisted Extraction, obtain lixiviating solution II, ultrasonic Assisted Extraction condition, power 400W, extraction temperature is 80 ℃, solid-liquid ratio is 1:15 (w/v), and extracting times is 2 times, and extraction time is 30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards.
(4) precipitate with ethanol processing: coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 95% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitation is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I.
(5) purification process: by final precipitate with ethanol precipitate I under room temperature by 80% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-60 ℃, be that 48h obtains Angelica Polysaccharide drying time.
(6) acid is heavy processes: adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Be concentrated into soluble solid to 35Brix, make concentration of alcohol lower than 5%.The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix.Use 1:1(v/v) aqueous sulfuric acid, stir and add, adjust pH to 2.0, obtain the heavy liquid I of acid.
(7) redissolve and process: acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved with organic solvent water saturation n-butyl alcohol, according to solid-to-liquid ratio 1:3(w/v), stirring and dissolving precipitation I 60min, makes it to dissolve completely, obtains organic solution I.
(8) ammonification processing: organic liquor I is adjusted to pH to 8.5 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I.
(9) acidification: use glacial acetic acid adjust pH to guarantee that to 3.0(acetic acid is excessive upper phase I), 80 ℃ of water bath heat preservation 30min, stand at low temperature 10h, obtains acidifying solution I.
(10) crystallization treatment: acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, stand at low temperature 8h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II.Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-60 ℃, and be 30h drying time; After precipitation is dry, obtain monoammonium glycyrrhizinate.
(11) purification process: supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, be concentrated into dry doubling 50% ethanol and redissolve, utilize AB-8 macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, the 10h that vibrates under eluent 80% ethanol room temperature carries out desorbing, collection eluent carries out vacuum evaporator and concentrates to remove ethanol, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, sprays dry.180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures, obtain licoflavone after being dried.
Monoammonium glycyrrhizinate yield 1.80% prepared by this condition, purity is 61.0%; Licoflavone yield 2.72%, purity is 29.4%; Angelica Polysaccharide yield 9.89%, purity is 59.3%.
Embodiment 5
A preparation method for Radix Glycyrrhizae active substance, its production stage is:
(1) pretreatment of raw material: Radix Glycyrrhizae is screened, removed various impurity, then washed, remove dust, dry, shattered 60 mesh sieves.
(2) hot water lixiviate processing: Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 95 ℃, and solid-liquid ratio is 1:10 (w/v), and extracting times is 3 times, and each extraction time is 3h.Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use.
(3) the ultrasonic Assisted Extraction processing of glycyrrhiza residue: the filtering residue of step (2) gained is used for to ultrasonic Assisted Extraction, obtain lixiviating solution II, ultrasonic Assisted Extraction condition, power 500W, extraction temperature is 80 ℃, solid-liquid ratio is 1:15 (w/v), and extracting times is 2 times, and extraction time is 30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards.
(4) precipitate with ethanol processing: coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 40Brix, obtain concentrated solution I, concentrated solution I adds 95% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 35Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5(w/v) with 75% ethanol redissolve, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I.
(5) purification process: by final precipitate with ethanol precipitate I under room temperature by 90% washing with alcohol 3 times, collecting precipitation, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-80 ℃, be that 48h obtains Angelica Polysaccharide drying time.
(6) acid is heavy processes: adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Be concentrated into soluble solid 30Brix, make concentration of alcohol lower than 5%.The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix.Use 1:1(v/v) aqueous sulfuric acid, stir and add, adjust diluent pH value to 1.5, obtain the heavy liquid I of acid.
(7) redissolve and process: acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved with organic solvent water saturation n-butyl alcohol, according to solid-to-liquid ratio 1:3(w/v), stirring and dissolving precipitation I 50min, makes it to dissolve completely, obtains organic liquor I.
(8) ammonification processing: organic solution I is adjusted to pH to 8.5 with strong aqua ammonia, magnetic agitation 1h, stratification, obtains phase I, lower phase I.
(9) acidification: use glacial acetic acid adjust pH to guarantee that to 3.0(acetic acid is excessive upper phase I), 90 ℃ of water bath heat preservation 30min, stand at low temperature 12h, obtains acidifying solution I.
(10) crystallization treatment: acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, stand at low temperature 8h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II.Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-60 ℃, and be 30h drying time; After drying precipitate, obtain monoammonium glycyrrhizinate.
(11) purification process: supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, being concentrated into dry doubling 50% ethanol redissolves, utilize XDB-8 macroporous resin to carry out purification, the 12h that vibrates under room temperature carries out static adsorption suction, and the 12h that vibrates under eluent 90% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry.200 ℃ of the dry inlet temperatures of spraying, 85 ℃ of outlet temperatures; After dry, obtain licoflavone.
Monoammonium glycyrrhizinate yield 1.92% prepared by this condition, purity is 62.1%; Licoflavone yield 3.30%, purity is 31.2%; Angelica Polysaccharide yield 10.61%, purity is 60.4%.

Claims (8)

1. a preparation method for Radix Glycyrrhizae active substance, comprising:
The pretreatment of raw material; Radix Glycyrrhizae is screened, removed various impurity, wash, be dried, pulverized 60 mesh sieves;
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I, and extraction temperature is 75-95 ℃, and solid-liquid ratio is 1:5-1:15, and extracting times is 1-3 time, and each extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue obtaining is for ultrasonic Assisted Extraction, obtain lixiviating solution II, ultrasonic power 300-500W, extraction temperature is 60-80 ℃, solid-liquid ratio is 1:10-1:20, and extracting times is 1-2 time, and each extraction time is 20-30min, lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing, coarse filtration liquid I is through centrifugal treating, rotating speed 4200r/min, centrifugal 10min, obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, being concentrated into soluble solid is 30-40Brix, obtain concentrated solution I, concentrated solution I adds 75-95% ethanol to make it ethanol final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 25-35Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75-95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; Final precipitate with ethanol precipitate I is used under room temperature to 60-90% washing with alcohol 1-3 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, and condenser temperature is-40 ℃~-80 ℃, and be 36-48h drying time, obtains Angelica Polysaccharide.
2. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 1, is characterized in that: also comprise the heavy step of processing, redissolving processing, ammonification processing, acidification and crystallization of acid, wherein,
Acid is heavy to be processed; Adopt vacuum evaporator concentrated to remove ethanol final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, vacuum is 0.08MPa, is concentrated into soluble solid for being concentrated into 30-35Brix, makes concentration of alcohol lower than 5%, obtain concentrated solution II, concentrated solution II is diluted and made soluble solid reach 15Brix with deionized water, use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 1.5-2.5, obtain the heavy liquid I of acid;
Redissolve and process; Heavy acid liquid I, in 4 ℃ of standing 12h, with rotating speed 4200r/min, is carried out to centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent I, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 30-60min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 7.5-9.0 with strong aqua ammonia, stir 1h, static layering, obtains phase I, lower phase I;
Acidification; Upper phase I is used below glacial acetic acid adjust pH to 4.0,70-90 ℃ of water bath heat preservation 30min, stand at low temperature 6-12h, obtains acidifying solution I;
Crystallization; Acidifying solution I adds dehydrated alcohol dilution, make concentration of alcohol reach 90%, stand at low temperature 4-8h, with the centrifugal 20min of 4200r/min, obtain supernatant II and precipitation after centrifugal, the precipitation after centrifugal is carried out vacuum lyophilization, vacuum is 0.014MPa, condenser temperature is-40 ℃~-60 ℃, and be 20-30h drying time, after precipitation is dry, obtains monoammonium glycyrrhizinate.
3. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 2, it is characterized in that: also comprise purification step, supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, being concentrated into dry doubling 50% ethanol redissolves, utilize macroporous resin to carry out purification, resin is selected XDA-8, LX20B, D101, AB-8 is wherein a kind of, the 8-12h that vibrates under room temperature carries out static adsorption, the 8-12h that vibrates under 70-90% ethanol room temperature carries out desorbing, it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, being concentrated into soluble solid is 30-35Brix, spray dry.The dry inlet temperature 160-200 ℃ of spraying, outlet temperature 80-85 ℃, obtains licoflavone after being dried.
4. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 1, is characterized in that:
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 75 ℃, and solid-liquid ratio is 1:5, and extracting times is 1 time, and extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained for ultrasonic Assisted Extraction, obtains lixiviating solution II, ultrasonic Assisted Extraction condition: power 300W, extraction temperature are that 60 ℃, solid-liquid ratio are that 1:10, extracting times are that 1 time, extraction time are 20min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing, coarse filtration liquid I process centrifugal treating, rotating speed 4200r/min, centrifugal 10min, obtains clear liquor I, and clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 30Brix, obtain concentrated solution I, concentrated solution I adds 75% ethanol to make it ethanol final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 25Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 60% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%, obtains concentrated solution II, concentrated solution II is diluted and made soluble solid reach 15Brix with deionized water, use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 1.5, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent ethyl acetate, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 30min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 7.5 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 4.0, guarantee that acetic acid is excessive, 70 ℃ of water bath heat preservation 30min, stand at low temperature 6h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 4h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-40 ℃, and be 20h drying time, obtains monoammonium glycyrrhizinate after drying precipitate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, and are concentrated into dry doubling 50% ethanol and redissolve.Utilize XDA-8 macroporous resin to carry out purification, the 8h that vibrates under room temperature carries out static adsorption suction, the 8h that vibrates under eluent 70% ethanol room temperature carries out desorbing, it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry; 160 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
5. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 1, is characterized in that:
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I, and extraction temperature is 85 ℃, and solid-liquid ratio is 1:15, and extracting times is 3 times, and each extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition: power 500W, extraction temperature are that 70 ℃, solid-liquid ratio are that 1:10, extracting times are that 2 times, extraction time are 30min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing; Coarse filtration liquid I process centrifugal treating, with rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 85% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate redissolves with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 70% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%; The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix; Use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 2.0, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent ethyl acetate, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 40min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 8.0 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.5,80 ℃ of water bath heat preservation 30min, stand at low temperature 8h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 6h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-50 ℃, and be 25h drying time; After drying precipitate, obtain monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, be concentrated into dry doubling 50% ethanol and redissolve, utilize macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, the 10h that vibrates under eluent 70% ethanol room temperature carries out desorbing, collection eluent carries out vacuum evaporator and concentrates to remove ethanol, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 30Brix, sprays dry.180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
6. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 1, is characterized in that:
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I, and extraction temperature is 85 ℃, and solid-liquid ratio is 1:10, and extracting times is 2 times, and each extraction time is 3h, and lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained, for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition, power 400W, extraction temperature is 70 ℃, and solid-liquid ratio is 1:20, and extracting times is 1 time, and extraction time is 30min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing; Coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 85% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate is for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 85% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 70% washing with alcohol 2 times, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-40 ℃, be that 36h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Being concentrated into soluble solid is 30Brix, makes concentration of alcohol lower than 5%, and the concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix; Use volume ratio 1:1 aqueous sulfuric acid, stir and add, adjust diluent pH value to 2.5, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved by organic solvent dichloromethane, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 40min, make it to dissolve completely, obtain organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 9.0 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.5,80 ℃ of water bath heat preservation 30min, stand at low temperature 8h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 6h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-50 ℃, and be 25h drying time, after precipitation is dry, obtains monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, vacuum concentration to dry doubling redissolves with 50% ethanol, utilize macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, and the 10h that vibrates under eluent 80% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry; 180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures; After dry, obtain licoflavone.
7. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 1, is characterized in that:
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I.Extraction temperature is 85 ℃, and solid-liquid ratio is 1:15, and extracting times is 2 times, and each extraction time is 3h; Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition: power 400W, extraction temperature are that 80 ℃, solid-liquid ratio are that 1:15, extracting times are that 2 times, extraction time are 30min; Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing, coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min, obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, obtain concentrated solution I, concentrated solution I adds 95% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 30Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 95% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitation is redissolved with 75% ethanol according to solid-to-liquid ratio 1:5, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 80% washing with alcohol 1 time, collecting precipitation thing, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-60 ℃, be that 48h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Be concentrated into soluble solid to 35Brix, make concentration of alcohol lower than 5%, the concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix; Use volume ratio 1:1 aqueous sulfuric acid to stir and add, adjust pH to 2.0, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved with organic solvent water saturation n-butyl alcohol, according to solid-to-liquid ratio 1:3 stirring and dissolving precipitation I 60min, make it to dissolve completely, obtain organic solution I;
Ammonification processing; Organic liquor I is adjusted to pH to 8.5 with strong aqua ammonia, stir 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.0,80 ℃ of water bath heat preservation 30min, stand at low temperature 10h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 8h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-60 ℃, and be 30h drying time; After precipitation is dry, obtain monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, be concentrated into dry doubling 50% ethanol and redissolve, utilize macroporous resin to carry out purification, the 10h that vibrates under room temperature carries out static adsorption suction, the 10h that vibrates under eluent 80% ethanol room temperature carries out desorbing, collection eluent carries out vacuum evaporator and concentrates to remove ethanol, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 35Brix, sprays dry; 180 ℃ of the dry inlet temperatures of spraying, 80 ℃ of outlet temperatures, obtain licoflavone after being dried.
8. a kind of preparation method of Radix Glycyrrhizae active substance as claimed in claim 1, is characterized in that:
Hot water lixiviate processing; Radix Glycyrrhizae hot water lixiviate processing, obtains lixiviating solution I; Extraction temperature is 95 ℃, and solid-liquid ratio is 1:10, and extracting times is 3 times, and each extraction time is 3h; Lixiviating solution I obtains coarse filtration liquid I by 200 order filter-cloth filterings, and filtering residue is for subsequent use;
The ultrasonic Assisted Extraction processing of glycyrrhiza residue; The filtering residue of gained, for ultrasonic Assisted Extraction, is obtained to lixiviating solution II, ultrasonic Assisted Extraction condition, power 500W, extraction temperature is 80 ℃, and solid-liquid ratio is 1:15, and extracting times is 2 times, and extraction time is 30min.Lixiviating solution II obtains coarse filtration liquid II by 200 order filter-cloth filterings, and filtering residue discards;
Precipitate with ethanol processing; Coarse filtration liquid I is through centrifugal treating rotating speed 4200r/min, centrifugal 10min.Obtain clear liquor I, clear liquor I is concentrated through vacuum evaporator, and thickening temperature is 45 ℃, and vacuum is 0.08MPa, being concentrated into soluble solid is 40Brix, obtain concentrated solution I, concentrated solution I adds 95% ethanol to make it final concentration to reach 50%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the first time, precipitate for subsequent use, precipitate with ethanol supernatant is concentrated into 35Brix by vacuum evaporator for the first time, carry out precipitate with ethanol for the second time, add 75% ethanol to make it final concentration and reach 70%, room temperature leaves standstill 4h, with the centrifugal 10min of 4200r/min, precipitate is according to solid-to-liquid ratio 1:5, with 75% ethanol redissolution, 90 ℃ of backflow 30min, centrifugal filtration, supernatant is incorporated to precipitate with ethanol supernatant for the second time and obtains final precipitate with ethanol supernatant I, the precipitate of twice ethanol redissolution merges, for final precipitate with ethanol precipitate I,
Purification process; By final precipitate with ethanol precipitate I under room temperature by 90% washing with alcohol 3 times, collecting precipitation, adopts vacuum freeze-drying technique to carry out dried, condenser temperature is-80 ℃, be that 48h obtains Angelica Polysaccharide drying time;
Acid is heavy to be processed; Adopt vacuum evaporator concentrated final precipitate with ethanol supernatant I, thickening temperature is 45 ℃, and vacuum is 0.08MPa; Be concentrated into soluble solid 30Brix, make concentration of alcohol lower than 5%; The concentrated solution deionized water that removes ethanol is diluted and makes soluble solid reach 15Brix.Use volume ratio 1:1 aqueous sulfuric acid to stir and add, adjust diluent pH value to 1.5, obtain the heavy liquid I of acid;
Redissolve and process; Acid is sunk to liquid I in 4 ℃ of standing 12h, with rotating speed 4200r/min, carry out centrifugalize 20min, obtain supernatant I and precipitation I, supernatant I is for subsequent use, and precipitation I is redissolved with organic solvent water saturation n-butyl alcohol, according to solid-to-liquid ratio 1:3, stirring and dissolving precipitation I 50min, makes it to dissolve completely, obtains organic liquor I;
Ammonification processing; Organic solution I is adjusted to pH to 8.5 with strong aqua ammonia, magnetic agitation 1h, stratification, obtains phase I, lower phase I;
Acidification; Upper phase I is used to glacial acetic acid adjust pH to 3.0,90 ℃ of water bath heat preservation 30min, stand at low temperature 12h, obtains acidifying solution I;
Crystallization treatment; Acidifying solution I adds dehydrated alcohol dilution, makes concentration of alcohol reach 90%, and stand at low temperature 8h, with the centrifugal 20min of 4200r/min, obtains supernatant II and precipitation II; Precipitation II is carried out vacuum lyophilization, and vacuum is 0.014MPa, and condenser temperature is-60 ℃, and be 30h drying time; After drying precipitate, obtain monoammonium glycyrrhizinate;
Purification process; Supernatant I, coarse filtration liquid II, lower phase I and supernatant II merge, being concentrated into dry doubling 50% ethanol redissolves, utilize macroporous resin to carry out purification, the 12h that vibrates under room temperature carries out static adsorption suction, and the 12h that vibrates under eluent 90% ethanol room temperature carries out desorbing, and it is concentrated to remove ethanol that collection eluent carries out vacuum evaporator, thickening temperature is 45 ℃, vacuum is 0.08MPa, and being concentrated into soluble solid is 30Brix, sprays dry; 200 ℃ of the dry inlet temperatures of spraying, 85 ℃ of outlet temperatures; After dry, obtain licoflavone.
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CN104387439A (en) * 2014-10-31 2015-03-04 中国农业大学 Method for preparing glycyrrhizic acid derivatives by carrying out subcritical hydrolysis reaction
CN104387439B (en) * 2014-10-31 2017-02-01 中国农业大学 Method for preparing glycyrrhizic acid derivatives by carrying out subcritical hydrolysis reaction
CN104861015A (en) * 2015-01-16 2015-08-26 李玉山 Synergistic clean extraction method of effective components in licorice root
CN104983778A (en) * 2015-06-29 2015-10-21 宁夏中汇天颐生物科技有限公司 Method for continuously and comprehensively extracting liquorice ingredient with high pressure
CN104983778B (en) * 2015-06-29 2020-03-31 宁夏中汇天颐生物科技有限公司 Method for continuously and comprehensively extracting liquorice components under high pressure
CN106279345A (en) * 2016-08-15 2017-01-04 晨光生物科技集团股份有限公司 A kind of industrialized preparing process of thick glycyrrhizic acid
CN106478828A (en) * 2016-09-30 2017-03-08 中国农业大学 A kind of method having anti-oxidation function polysaccharide powder in ultrasound assisted extraction Abelmoschus esculentus
CN107495459A (en) * 2017-10-17 2017-12-22 江西中烟工业有限责任公司 The additive of harmful components in a kind of reduction smoke of tobacco
CN109666085A (en) * 2019-01-09 2019-04-23 广州市莱檬生物科技有限公司 A method of pectin from lemon peel is purified using alcohol precipitation
CN109666085B (en) * 2019-01-09 2020-03-24 广州市莱檬生物科技有限公司 Method for purifying lemon peel pectin by alcohol precipitation
CN111607014A (en) * 2020-06-09 2020-09-01 佛山科学技术学院 Preparation method of glycyrrhiza polysaccharide iron chelate
CN114478680A (en) * 2021-12-15 2022-05-13 贵州弘康药业有限公司 Licorice extraction process

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