CN110846354A - Preparation method of blackberry lily isoflavone aglycone - Google Patents
Preparation method of blackberry lily isoflavone aglycone Download PDFInfo
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- CN110846354A CN110846354A CN201911199028.1A CN201911199028A CN110846354A CN 110846354 A CN110846354 A CN 110846354A CN 201911199028 A CN201911199028 A CN 201911199028A CN 110846354 A CN110846354 A CN 110846354A
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- blackberry lily
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- fibrous root
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- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 title claims abstract description 36
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 title claims abstract description 36
- 235000008696 isoflavones Nutrition 0.000 title claims abstract description 36
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 title claims abstract description 35
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- 238000002360 preparation method Methods 0.000 title claims abstract description 16
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- OYUJPVCKGSEYDD-UHFFFAOYSA-N tectorigenin Natural products COc1c(O)cc2OCC(C(=O)c2c1O)c1ccc(O)cc1 OYUJPVCKGSEYDD-UHFFFAOYSA-N 0.000 description 4
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- AEDDIBAIWPIIBD-ZJKJAXBQSA-N mangiferin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C1=C(O)C=C(OC=2C(=CC(O)=C(O)C=2)C2=O)C2=C1O AEDDIBAIWPIIBD-ZJKJAXBQSA-N 0.000 description 2
- 208000026435 phlegm Diseases 0.000 description 2
- 150000008442 polyphenolic compounds Chemical class 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- FHFSSMDJUNVMNY-UHFFFAOYSA-N tectoridin Natural products COc1c(O)c2C(=O)C(=COc2cc1OC3OC(CO)C(O)C(O)C3O)c4cccc(O)c4 FHFSSMDJUNVMNY-UHFFFAOYSA-N 0.000 description 2
- CNOURESJATUGPN-UDEBZQQRSA-N tectoridin Chemical compound C1=C2OC=C(C=3C=CC(O)=CC=3)C(=O)C2=C(O)C(OC)=C1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O CNOURESJATUGPN-UDEBZQQRSA-N 0.000 description 2
- QDVIEIMMEUCFMW-QXYPORFMSA-N (3z,5e)-6-(3,4-dihydroxyphenyl)-4-hydroxyhexa-3,5-dien-2-one Chemical compound CC(=O)\C=C(/O)\C=C\C1=CC=C(O)C(O)=C1 QDVIEIMMEUCFMW-QXYPORFMSA-N 0.000 description 1
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 1
- 241000222501 Agaricaceae Species 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 241000123392 Hymenochaetaceae Species 0.000 description 1
- 241000123247 Inonotus Species 0.000 description 1
- 241000123249 Inonotus hispidus Species 0.000 description 1
- 102000003777 Interleukin-1 beta Human genes 0.000 description 1
- 108090000193 Interleukin-1 beta Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 241001113425 Iridaceae Species 0.000 description 1
- YWQSXCGKJDUYTL-UHFFFAOYSA-N Mangiferin Natural products CC(CCC=C(C)C)C1CC(C)C2C3CCC4C(C)(C)CCCC45CC35CCC12C YWQSXCGKJDUYTL-UHFFFAOYSA-N 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
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- 210000005064 dopaminergic neuron Anatomy 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- SGJNQVTUYXCBKH-HNQUOIGGSA-N hispidin Chemical compound O1C(=O)C=C(O)C=C1\C=C\C1=CC=C(O)C(O)=C1 SGJNQVTUYXCBKH-HNQUOIGGSA-N 0.000 description 1
- SGJNQVTUYXCBKH-UHFFFAOYSA-N hispidin Natural products O1C(=O)C=C(O)C=C1C=CC1=CC=C(O)C(O)=C1 SGJNQVTUYXCBKH-UHFFFAOYSA-N 0.000 description 1
- QDVIEIMMEUCFMW-UHFFFAOYSA-N hispolone Natural products CC(=O)C=C(O)C=CC1=CC=C(O)C(O)=C1 QDVIEIMMEUCFMW-UHFFFAOYSA-N 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229940043357 mangiferin Drugs 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
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- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
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- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
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- 108090000623 proteins and genes Proteins 0.000 description 1
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- 239000010409 thin film Substances 0.000 description 1
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- 238000013518 transcription Methods 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of traditional Chinese medicine preparation, and particularly discloses a preparation method of blackberry lily isoflavone aglycone. The preparation method of the blackberry lily isoflavone aglycone comprises the steps of carrying out alcohol removal treatment on an alcohol extract of blackberry lily fibrous roots, then adding the alcohol extract into a fermentation liquor of crude capillary fungus for enzymolysis, removing enzyme and thallus impurities in the enzymolysis liquor, and carrying out resin adsorption and purification to obtain the blackberry lily isoflavone aglycone. The method has the advantages of mild preparation conditions, simple method and low cost, and greatly improves the purity and yield of the blackberry lily isoflavone aglycone extracted and prepared from the blackberry lily fibrous root.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine preparation, in particular to a preparation method of blackberry lily isoflavone aglycone.
Background
Belamcanda chinensis is the dry rhizome of Iridaceae plant, is bitter in taste and cold in nature, enters liver and lung channels, has the effects of clearing heat and removing toxicity, relieving sore throat and eliminating phlegm, is mainly used for treating throat lung carbuncle, phlegm cough and asthma and the like, and is the essential medicine for treating pharyngitis and sore throat.
The research shows that the tectorigenin and other aglycone substances can obviously inhibit the transcription and translation levels of tumor immunocytes TNF- α, IL-1 β and IL-6, the tectorigenin and other aglycone substances can also prevent the accumulation of α -synuclein, slow down the degenerative change of dopaminergic neurons caused by hydroxypolyamide, enhance the food sensitivity and prolong the service life, the compound also has the treatment effect on potential gold-induced diseases, the activity of the isoflavone substances such as the tectorigenin, the wild tectorigenin and other isoflavone substances and the like does not waste a large amount of isoflavone resources in extraction, and the extraction cost is low, so that the extraction cost is low, and the extraction cost is low.
Disclosure of Invention
Aiming at the problems of low extraction rate and low product purity of the existing method for separating and extracting isoflavone aglycone from the fibrous root of the blackberry lily medicinal material, the invention provides a preparation method of the blackberry lily isoflavone aglycone.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
a method for preparing rhizoma Belamcandae isoflavone aglycone comprises removing alcohol from rhizoma Belamcandae fibrous root alcoholic extractive solution, adding into crude Fibrouromycetes fermentation broth for enzymolysis, removing enzyme and thallus impurities in the enzymolysis solution, and performing resin adsorption and purification to obtain rhizoma Belamcandae isoflavone aglycone.
Wherein, the crude capillary is medicinal fungus crude capillary [ Inonotus hispidus (Bull.: Fr.) P.Karst. ], the name Xanthochrous fuscus (Xanthochrous hispidus) is superior to Basidiomycota (Basidiomycota), Agaricaceae (Agaricamycetes), Hymenochaetales (Hymenochaetales), Inonochaetaceae (Hymenochaetaceae), and Microchaetaceae (Inonotus); the strain is mainly used for treating various cancers, arthritis, gout, diabetes and other diseases and various stomach diseases caused by dyspepsia and the like, and the crude capillary fungus mainly contains polyphenol, triterpene, sterol and derivatives thereof, nucleoside, polysaccharide and other active ingredients, wherein polyphenol compounds Hispidin and Hispolon have various pharmacological actions of resisting virus, resisting tumor, resisting oxidation and the like.
Compared with the prior art, in the preparation method of blackberry lily isoflavone aglycone, isoflavone glycoside substances extracted from blackberry lily fibrous root alcohol extract are hydrolyzed by using various degrading enzymes generated in the fermentation process of crude capillary, so that various isoflavone glycoside substances (mangiferin, tectoridin, wild tectoridin and the like) which are difficult to degrade and have low biological activity in the alcohol extract can be fully hydrolyzed and converted to form isoflavone aglycone, the yield of the isoflavone aglycone extracted and prepared from blackberry lily fibrous root is greatly increased, the alcohol extract is subjected to resin adsorption and purification after enzymolysis, high-purity isoflavone aglycone can be obtained, the preparation condition is mild, the preparation method is simple, the cost is low, and the method is suitable for industrial production.
Preferably, the blackberry lily fibrous root ethanol extract is obtained by crushing blackberry lily fibrous roots, adding the crushed blackberry lily fibrous roots into 60-80 vt% ethanol solution, and leaching at 50-70 ℃.
Preferably, the blackberry lily roots are crushed to the granularity of 20-40 meshes.
Preferably, the leaching process is carried out by two times, in the first leaching process, the mass ratio of the blackberry lily fibrous root to the ethanol solution is 1:12-14, and the leaching time is 15-30min at 1000r/min of 800-; in the second leaching process, the mass ratio of the blackberry lily fibrous root to the ethanol solution is 1:8-10, and the leaching is carried out for 15-30min at the speed of 1000r/min of 800-; and combining the two leaching solutions to obtain the alcoholic leaching solution of the blackberry lily fibrous roots.
The above alcohol extraction process can extract isoflavone glycoside contained in rhizoma Belamcandae.
Preferably, the dealcoholization treatment process is to filter the alcoholic extract of the dried fibrous roots and concentrate the alcoholic extract by a thin film evaporator under reduced pressure until the alcohol content is less than or equal to 5vt percent.
The extraction liquid is subjected to alcohol removal treatment, so that the influence of alcohol on the hydrolysis efficiency of enzyme in the subsequent enzymolysis process can be avoided.
Preferably, the fermentation liquid of the crude capillary is a homogenate enzyme liquid obtained by inoculating the crude capillary into a culture solution, culturing for 7-10 days at 28-30 ℃, and breaking the cell wall of the culture solution by using a homogenizer.
The wall breaking treatment can fully release intracellular enzymes in the thalli, increase the concentration and the variety of the enzymes and improve the enzymolysis efficiency.
Preferably, the addition volume of the fermentation liquid of the crude phellinus linteus is 5 to 10% of the volume of the concentrated liquid after the dealcoholization treatment.
Preferably, the enzymolysis process is as follows: performing enzymolysis at 40-55 deg.C and pH of 4-6 for 1-3 days.
Under the enzymolysis condition, the hydrolysis efficiency of the enzyme in the enzymolysis liquid to the isoflavone glycoside substances can be improved.
Preferably, the culture solution comprises 0.6-0.8% of malt extract, 0.07-0.08% of yeast extract, 0.8-1.2% of peptone and the balance of distilled water by mass ratio.
Preferably, the process for removing the enzyme and the bacterial impurities in the enzymatic hydrolysate comprises the following steps: and adding a flocculating agent into the enzymolysis liquid, precipitating for 2-3h, and carrying out centrifugal filtration to obtain a clear liquid.
The impurity removal method can fully remove enzyme, other macromolecular protein and thallus fragments in the enzymolysis liquid, and ensures the smooth proceeding of the subsequent resin adsorption treatment.
Preferably, the flocculant is one or more of chitosan, gelatin and A + B clarifying agent; the addition amount of the flocculating agent is 3-5% of the mass of the enzymolysis liquid.
Preferably, the resin is macroporous adsorption resin, and specifically one of HPD700, LX-17 and LK-17 macroporous resin can be selected.
Preferably, the resin adsorption purification process is as follows: diluting the enzymolysis solution by 1-1.5 times, adding into macroporous adsorption resin, controlling the adsorption flow rate at 1-1.5BV/h, eluting with 20-40 vt% ethanol solution to remove impurities, eluting with 60-80 vt% ethanol solution, concentrating and drying.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The following examples are provided to better illustrate the embodiments of the present invention.
Example 1
A preparation method of blackberry lily isoflavone aglycone comprises the following steps:
pretreatment of raw materials: crushing 1kg of blackberry lily fibrous roots into powder with the particle size of 20 meshes;
alcohol extraction: weighing the crushed blackberry lily fibrous root raw material, stirring and extracting twice by using 60% ethanol in a water bath at 50 ℃, wherein in the first extraction process, the blackberry lily fibrous root and an ethanol solution are extracted for 15min at a mass ratio of 1:12 and at a speed of 800 r/min; in the second leaching process, the mass ratio of the blackberry lily fibrous root subjected to the first leaching to the ethanol solution is 1:8, and the leaching is carried out for 15min at the speed of 800 r/min;
concentrating and removing alcohol: mixing and filtering the two alcohol extract solutions, and then carrying out reduced pressure concentration by using a film evaporator until the alcohol extract solution reaches an alcohol-free state;
enzymolysis: inoculating medicinal fungus crude capillary fungus into a culture solution, culturing for 7 days at 28 ℃, and performing wall breaking and homogenizing treatment on thalli in the culture solution by using a homogenizer to obtain a homogenized enzyme solution, wherein the culture solution comprises 0.6% of malt extract, 0.07% of yeast extract, 0.8% of peptone and the balance of distilled water according to the mass ratio; adding the concentrated alcohol extract solution after removing alcohol into the homogenate, wherein the volume of the concentrated solution is 5% of that of the alcohol extract solution, and performing enzymolysis for 1 day at 40 ℃ and pH 4;
precipitation and filtration: adding chitosan into the enzymolysis liquid, wherein the adding amount of the chitosan is 3% of the mass of the enzymolysis liquid, performing flocculation precipitation for 2 hours, and performing centrifugal filtration to obtain clear liquid;
resin adsorption and purification: diluting the clear liquid by 1 time, adding the diluted clear liquid into HPD700 macroporous adsorption resin, controlling the adsorption flow rate to be 1BV/h, after the adsorption is finished, eluting and removing impurities by using 20 vt% ethanol solution, eluting, vacuum-concentrating and drying by using 60 vt% ethanol solution to obtain 22g of isoflavone aglycone product;
the isoflavone aglycone content in the obtained isoflavone aglycone product was 77.1%.
Example 2
A preparation method of blackberry lily isoflavone aglycone comprises the following steps:
pretreatment of raw materials: crushing 1kg of blackberry lily fibrous roots into powder with the particle size of 30 meshes;
alcohol extraction: weighing the sheared or crushed blackberry lily fibrous root raw material, stirring and extracting the blackberry lily fibrous root raw material twice by using 70% ethanol in a water bath at the temperature of 60 ℃, wherein in the first extraction process, the blackberry lily fibrous root and ethanol solution are extracted for 20min at the mass ratio of 1:13 and 900 r/min; in the second leaching process, the mass ratio of the blackberry lily fibrous root subjected to the first leaching to the ethanol solution is 1:9, and the leaching is carried out for 20min at 900 r/min;
concentrating and removing alcohol: mixing and filtering the two alcohol extract solutions, and then carrying out reduced pressure concentration by using a film evaporator until the alcohol extract solution reaches an alcohol-free state;
enzymolysis: inoculating medicinal fungus crude capillary fungus into a culture solution, culturing for 8 days at 29 ℃, and performing wall breaking and homogenizing treatment on thalli in the culture solution by using a homogenizer to obtain a homogenized enzyme solution, wherein the culture solution comprises 0.7% of malt extract, 0.075% of yeast extract, 1% of peptone and the balance of distilled water in mass ratio; adding the concentrated alcohol extract solution after removing alcohol into the homogenate, wherein the volume of the concentrated solution is 8% of the volume of the alcohol extract solution, and performing enzymolysis for 2 days at 50 ℃ and pH 5;
precipitation and filtration: adding gelatin into the enzymolysis liquid, wherein the adding amount of the gelatin is 4% of the mass of the enzymolysis liquid, performing flocculation precipitation for 2.5h, and performing centrifugal filtration to obtain clear liquid;
resin adsorption and purification: diluting the clear liquid by 1 time, adding the diluted clear liquid into LX-17 macroporous adsorption resin, controlling the adsorption flow rate to be 1.2BV/h, eluting and removing impurities by using 30 vt% ethanol solution after adsorption is finished, eluting, vacuum concentrating and drying by using 70 vt% ethanol solution to obtain 21g of isoflavone aglycone product;
the isoflavone aglycone content in the obtained isoflavone aglycone product is 79.5 percent.
Example 3
A preparation method of blackberry lily isoflavone aglycone comprises the following steps:
pretreatment of raw materials: crushing 1kg of blackberry lily fibrous roots into powder with the particle size of 40 meshes;
alcohol extraction: weighing the sheared or crushed blackberry lily fibrous root raw material, stirring and extracting the blackberry lily fibrous root raw material twice by using 80% ethanol in a water bath at 70 ℃, wherein in the first extraction process, the weight ratio of blackberry lily fibrous root to ethanol solution is 1:14, and the extraction time is 30min at 1000 r/min; in the second leaching process, the mass ratio of the blackberry lily fibrous root subjected to the first leaching to the ethanol solution is 1:10, and the leaching is carried out for 30min at 1000 r/min;
concentrating and removing alcohol: mixing and filtering the two alcohol extract solutions, and then carrying out reduced pressure concentration by using a film evaporator until the alcohol extract solution reaches an alcohol-free state;
enzymolysis: inoculating medicinal fungus crude capillary fungus into a culture solution, culturing for 10 days at 30 ℃, and performing wall breaking and homogenizing treatment on thalli in the culture solution by using a homogenizer to obtain a homogenized enzyme solution, wherein the culture solution comprises 0.8% of malt extract, 0.08% of yeast extract, 1.2% of peptone and the balance of distilled water according to the mass ratio; adding the concentrated alcohol extract solution after removing alcohol into the homogenate, wherein the volume of the concentrated solution is 10 percent of the volume of the alcohol extract solution, and performing enzymolysis for 3 days at the temperature of 55 ℃ and the pH value of 6;
precipitation and filtration: adding an A + B clarifying agent into the enzymolysis liquid, wherein the addition amount of the A + B clarifying agent is 5% of the mass of the enzymolysis liquid, performing flocculation precipitation for 3 hours, and performing centrifugal filtration to obtain a clear liquid;
resin adsorption and purification: diluting the clear liquid by 1 time, adding the diluted clear liquid into LK-17 macroporous adsorption resin, controlling the adsorption flow rate to be 1.5BV/h, after the adsorption is finished, firstly eluting by using 40 vt% ethanol solution to remove impurities, then eluting by using 80 vt% ethanol solution, concentrating in vacuum and drying to obtain 24g isoflavone aglycone product;
the isoflavone aglycone content in the obtained isoflavone aglycone product is 80.2%.
Comparative example 1
The enzymolysis process in the embodiment 1 is removed, and the concentrated alcohol extract without alcohol is directly subjected to resin adsorption purification;
16g of isoflavone aglycone product with an isoflavone aglycone content of 53.1% was finally obtained.
Comparative example 2
β -glucosidase is used for replacing the fungus crude capillary fungus fermentation broth homogenate in the embodiment 1, and the alcohol-removed blackberry lily fibrous root alcohol extract is subjected to enzymolysis, wherein the specific enzymolysis method is that β -glucosidase is added into the alcohol-removed blackberry lily fibrous root alcohol extract, the adding amount of β -glucosidase is 4000U/g blackberry lily fibrous root raw material, and the enzymolysis is carried out for 1 day under the conditions of 40 ℃ and pH 5;
the other procedures were the same as in example 1 to obtain 18g of an isoflavone aglycone product having an isoflavone aglycone content of 61.8%.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A preparation method of blackberry lily isoflavone aglycone is characterized by comprising the following steps: removing alcohol from rhizoma Belamcandae fibrous root alcoholic extractive solution, adding into crude capillary fungus fermentation broth for enzymolysis, removing enzyme and thallus impurities in the enzymolysis solution, and performing resin adsorption and purification to obtain rhizoma Belamcandae isoflavone aglycone.
2. The method of claim 1, wherein: the alcoholic extract of the blackberry lily fibrous root is obtained by crushing the blackberry lily fibrous root, adding the crushed blackberry lily fibrous root into 60-80 vt% ethanol solution, and leaching at 50-70 ℃.
3. The method of claim 2, wherein: crushing the blackberry lily fibrous roots to 20-40 meshes of granularity; and/or
The leaching process is carried out by two times, in the first leaching process, the mass ratio of the blackberry lily fibrous root to the ethanol solution is 1:12-14, and the leaching time is 15-30min at the speed of 1000 r/min; in the second extraction process, the mass ratio of the blackberry lily fibrous root to the ethanol solution is 1:8-10, and the extraction is carried out for 15-30min at the speed of 1000r/min 800-.
4. The method of claim 1, wherein: the alcohol removing treatment process is to filter and evaporate and concentrate the alcohol extract until the alcohol content in the alcohol extract is less than or equal to 5vt percent.
5. The method of claim 1, wherein: the fermentation liquor of the crude capillary is homogenate enzyme liquid obtained by inoculating the crude capillary into a culture solution, culturing for 7-10 days at 28-30 ℃, and breaking the cell wall of the thallus in the culture solution; and/or
The adding volume of the fermentation liquor of the crude phellinus linteus is 5-10% of the volume of the concentrated solution after the alcohol removal treatment; and/or
The enzymolysis process comprises the following steps: performing enzymolysis at 40-55 deg.C and pH of 4-6 for 1-3 days.
6. The method of claim 5, wherein: the culture solution comprises 0.6-0.8% of malt extract, 0.07-0.08% of yeast extract, 0.8-1.2% of peptone and the balance of distilled water according to the mass ratio.
7. The method of claim 1, wherein: the process for removing enzyme and thallus impurities in the enzymolysis liquid comprises the following steps: and adding a flocculating agent into the enzymolysis liquid, precipitating for 2-3h, and carrying out centrifugal filtration to obtain a clear liquid.
8. The method of claim 7, wherein: the flocculant is one or a combination of chitosan, gelatin and A + B clarifying agent; the addition amount of the flocculating agent is 3-5% of the mass of the enzymolysis liquid.
9. The method of claim 1, wherein: the resin is macroporous adsorption resin.
10. The method of claim 9, wherein: the resin adsorption purification process comprises the following steps: diluting the enzymolysis solution by 1-1.5 times, adding into macroporous adsorption resin, controlling the adsorption flow rate at 1-1.5BV/h, eluting with 20-40 vt% ethanol solution to remove impurities, eluting with 60-80 vt% ethanol solution, concentrating and drying.
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