CN111560409A - Preparation method of icariine - Google Patents
Preparation method of icariine Download PDFInfo
- Publication number
- CN111560409A CN111560409A CN202010383284.2A CN202010383284A CN111560409A CN 111560409 A CN111560409 A CN 111560409A CN 202010383284 A CN202010383284 A CN 202010383284A CN 111560409 A CN111560409 A CN 111560409A
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- China
- Prior art keywords
- mass
- extraction
- enzymolysis
- icariin
- epimedium
- Prior art date
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- TZJALUIVHRYQQB-XLRXWWTNSA-N icariin Chemical compound C1=CC(OC)=CC=C1C1=C(O[C@H]2[C@@H]([C@H](O)[C@@H](O)[C@H](C)O2)O)C(=O)C2=C(O)C=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C(CC=C(C)C)=C2O1 TZJALUIVHRYQQB-XLRXWWTNSA-N 0.000 title claims abstract description 97
- TZJALUIVHRYQQB-UHFFFAOYSA-N icariine Natural products C1=CC(OC)=CC=C1C1=C(OC2C(C(O)C(O)C(C)O2)O)C(=O)C2=C(O)C=C(OC3C(C(O)C(O)C(CO)O3)O)C(CC=C(C)C)=C2O1 TZJALUIVHRYQQB-UHFFFAOYSA-N 0.000 title claims abstract description 97
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 83
- 239000002994 raw material Substances 0.000 claims abstract description 57
- 238000000034 method Methods 0.000 claims abstract description 48
- TZJALUIVHRYQQB-XFDQAQKOSA-N Icariin Natural products O(C)c1ccc(C2=C(O[C@H]3[C@@H](O)[C@H](O)[C@@H](O)[C@H](C)O3)C(=O)c3c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O4)c(C/C=C(\C)/C)c3O2)cc1 TZJALUIVHRYQQB-XFDQAQKOSA-N 0.000 claims abstract description 46
- 241000893536 Epimedium Species 0.000 claims abstract description 30
- 235000018905 epimedium Nutrition 0.000 claims abstract description 29
- 229930002875 chlorophyll Natural products 0.000 claims abstract description 24
- 235000019804 chlorophyll Nutrition 0.000 claims abstract description 24
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 claims abstract description 24
- 102000004190 Enzymes Human genes 0.000 claims abstract description 21
- 108090000790 Enzymes Proteins 0.000 claims abstract description 21
- 239000011347 resin Substances 0.000 claims abstract description 21
- 229920005989 resin Polymers 0.000 claims abstract description 21
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- 150000004676 glycans Chemical class 0.000 claims abstract description 20
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- 238000001179 sorption measurement Methods 0.000 claims abstract description 17
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 13
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 44
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 36
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 30
- 238000000926 separation method Methods 0.000 claims description 27
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 24
- 239000012452 mother liquor Substances 0.000 claims description 24
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 22
- 239000001110 calcium chloride Substances 0.000 claims description 22
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 22
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 22
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 22
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- 239000000284 extract Substances 0.000 claims description 19
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical group [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 18
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- 229910021529 ammonia Inorganic materials 0.000 claims description 18
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- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 claims description 12
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- 238000005374 membrane filtration Methods 0.000 abstract description 3
- 238000004065 wastewater treatment Methods 0.000 abstract description 2
- 150000002500 ions Chemical class 0.000 abstract 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 21
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 16
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- HWDGVJUIHRPKFR-UHFFFAOYSA-I copper;trisodium;18-(2-carboxylatoethyl)-20-(carboxylatomethyl)-12-ethenyl-7-ethyl-3,8,13,17-tetramethyl-17,18-dihydroporphyrin-21,23-diide-2-carboxylate Chemical compound [Na+].[Na+].[Na+].[Cu+2].N1=C(C(CC([O-])=O)=C2C(C(C)C(C=C3C(=C(C=C)C(=C4)[N-]3)C)=N2)CCC([O-])=O)C(=C([O-])[O-])C(C)=C1C=C1C(CC)=C(C)C4=N1 HWDGVJUIHRPKFR-UHFFFAOYSA-I 0.000 description 7
- 229940079841 sodium copper chlorophyllin Drugs 0.000 description 7
- 235000013758 sodium copper chlorophyllin Nutrition 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
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- 150000002338 glycosides Chemical class 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
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- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
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- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
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- NGMYNFJANBHLKA-SENBMHEBSA-N Icariside II Natural products O(C)c1ccc(C2=C(O[C@H]3[C@@H](O)[C@H](O)[C@@H](O)[C@H](C)O3)C(=O)c3c(O)cc(O)c(C/C=C(\C)/C)c3O2)cc1 NGMYNFJANBHLKA-SENBMHEBSA-N 0.000 description 1
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- 239000005642 Oleic acid Substances 0.000 description 1
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- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
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- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07G—COMPOUNDS OF UNKNOWN CONSTITUTION
- C07G5/00—Alkaloids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/06—Benzopyran radicals
- C07H17/065—Benzo[b]pyrans
- C07H17/07—Benzo[b]pyran-4-ones
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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Abstract
The invention relates to a preparation method of icariin. The method solves the problems of low yield, low recovery rate of effective components, high cost, complex process, large wastewater treatment capacity, environmental pollution, difficult industrial production and the like in the prior method. The method comprises the steps of supercritical extraction, enzymolysis, adsorption, resolution, ion resin purification and membrane filtration, wherein a complex enzyme enzymolysis process is adopted to convert high-grade glucoside in epimedium into icariin as much as possible, the content of the icariin in the raw materials is increased to the maximum extent, the extraction rate is improved, the recovery rate of effective components reaches more than 90%, and the active components in the epimedium raw materials are developed to the maximum extent through the supercritical extraction, the ion resin purification and the membrane filtration to obtain chlorophyll, epimedium polysaccharide and alkaloid, so that the production cost of a single product is greatly reduced, and the production efficiency is greatly improved.
Description
Technical Field
The invention relates to a preparation method of icariin.
Background
Herba Epimedii is perennial herb of Epimedium of berberidaceae, and the medicinal part is dry leaf. The Chinese medicine has more than 2000 years of medicinal history, and is mainly distributed in Shanxi, Gansu, Shanxi, Henan, Qinghai, Hubei, Sichuan and other areas of China. Herba Epimedii is a traditional Chinese medicine tonic, and can be used for treating sexual impotence, premature ejaculation, soreness of waist, leg pain, numbness of limbs, hemiplegia, neurasthenia, amnesia, tinnitus, and blurred vision. The herba Epimedii contains flavone and polysaccharide as main ingredients, and also contains alkaloid, lignanoid, anthraquinone, volatile oil, stearic acid, linolenic acid, oleic acid, etc.
The Chinese pharmacopoeia stipulates that the main component of epimedium is icariin, the content of the raw material is not less than 0.5 percent, and the icariin can effectively improve the cardiovascular system and adjust the internal secretion; can also obviously reduce bone loss and increase the bone density of osteoporosis rats.
In addition, the epimedium also contains rich chlorophyll, polysaccharide, alkaloid and other effective components. Chlorophyll, a major pigment of plants for photosynthesis, has various uses such as hematopoiesis, vitamin provision, detoxification, disease resistance and the like. However, chlorophyll is not stable, and is decomposed by light, acid, alkali, oxygen, an oxidizing agent and the like, so that the chlorophyll is mostly synthesized into sodium copper chlorophyllin, so that the stability and the water solubility are improved, and the application range is widened. The epimedium polysaccharide can control cell division and differentiation, regulate cell growth and senility, and has obvious functions of resisting tumor, resisting virus, reducing blood fat, strengthening immunity, etc. and excellent development foreground. The alkaloid has various biological effects, can tonify kidney yang, strengthen bones and muscles, dispel wind-damp, and has good application prospect.
Looking up the data and finding, icariin among the prior art mostly directly adopts organic reagent, chromatographic column separation or enzymolysis purification after obtaining, the technology is complicated, with high costs, and the yield is low, difficult industrial production, and polysaccharide and alkaloid that remain in the mother liquor are handled as the discarded object, cause the waste in a large number of resources to cause extremely pollution to the environment, brought very big work load for sewage treatment, increased manufacturing cost.
Patent CN1225470C discloses a method for extracting icariine from Epimedium plant, which adopts aqueous organic solvent extraction, column chromatography and recrystallization to obtain 98% icariine, and the recovery rate of effective components is about 88%. The method adopts macroporous column chromatography and then silica gel column separation for recrystallization, has complex process, long period, large organic solvent consumption and low recovery rate of effective components, leads to high production cost and is not a good choice for industrial production.
In patent CN1268759C, icariin or aglycone is obtained by enzymolysis. Fermenting to obtain enzymolysis solution, adding ammonium sulfate and ethanol extraction enzyme, icariin, acetic acid or phosphoric acid, and reacting. The method has strong theory and weak practice, the product after enzymolysis is complex, high content icariine is difficult to obtain after silica gel column chromatography, the yield is low, and the method is not suitable for industrial production.
Patent CN101845067B relates to a preparation method of high-content icariin. Firstly, dissolving 15-40% of epimedium extract by adopting high-concentration ethanol, dissolving residue by adopting low-concentration ethanol, standing for crystallization, and recrystallizing to obtain 98% of icariin. Although the process is simple, the higher glucoside in the raw materials is not converted into icariin, which causes a great deal of waste of the raw materials and further causes high cost.
Patent CN103910772B mentions a method for extracting icariin. Extracting with ethanol, acidifying, separating with macroporous resin, separating with simulated moving bed chromatography, crystallizing to obtain icariin, subjecting the mother liquor to enzymolysis with Tween 80 and glucosidase or cellulase, and separating and purifying with chromatographic column filled with D101 and AB-8 macroporous resin to obtain baohuoside I, II. Although this process provides a short cycle route, the higher glycosides in the starting materials are likewise not converted to icariins and no specific yield is given. The simulated moving bed chromatography is used for separation in the purification process, the equipment is hardly applied to industrial production, and the operation is complex, the cost is high, and the equipment is not suitable for industrial production.
Patent CN106831913A provides a preparation method of icariin. Extracting raw materials with ethanol, concentrating, extracting with ethyl acetate or n-butanol, washing with anhydrous methanol or anhydrous ethanol or 40-95% ethanol or 30-95% methanol, and crystallizing the residue to obtain icariine with purity of over 98%. The recovery rate of the effective components is only 89 percent, the recovery rate is low, and the high-grade glycoside is not converted, thereby causing the waste of raw materials.
Patent CN110343731A adopts cellulase to hydrolyze herba Epimedii extract, the addition amount is 0.1-1% of the raw material mass, the obtained product after hydrolysis is herba Epimedii flavone low glycoside, no icariine is obtained, but directly enzymolysis is carried out to obtain secondary glycoside. The conversion rate is 97%, and the content of the glucoside is 86%.
Patent CN110343731A relates to a process for obtaining icariine by an enzymolysis method. The raw material is epimedium wushanense, and icariin with the content of more than 98 percent can be obtained by ethanol extraction, rhamnosidase enzymolysis, ethyl acetate extraction, crystallization and recrystallization. The overall yield was 81.2%. Although the conversion is high, the overall yield is low.
Disclosure of Invention
The invention aims to provide a preparation method of icariin with low cost and high recovery rate, which solves the problems of low yield, low recovery rate of effective components, high cost, complex process, large wastewater treatment capacity, environmental pollution, no contribution to industrial production and the like in the method. The invention enriches polysaccharide and alkaloid simultaneously while separating and purifying icariin to obtain a series of products with commercial value, realizes the maximization of resource utilization, reduces energy consumption and sewage treatment, and obviously reduces production cost.
The technical scheme of the invention is to provide a preparation method of icariin, which comprises the following steps:
pulverizing herba Epimedii raw material, extracting herba Epimedii raw material with weakly alkaline organic solvent as entrainer by supercritical carbon dioxide extraction method to obtain extractive solution and extraction residue, wherein the extractive solution is chlorophyll;
step 2, extracting and performing enzymolysis;
step 2.1, cooking the extraction residue obtained in the step 1 with alkaline water, then adding activated carbon, mixing uniformly, adding ethanol, stirring and extracting at normal temperature, filtering, and concentrating the filtrate;
step 2.2, adding a mixed reagent of calcium chloride and magnesium sulfate into the filtrate obtained in the step 2.1, stirring, filtering, adding rhamnosidase and xylosidase into the filtrate for enzymolysis, wherein the enzymolysis temperature is 45-55 ℃, inactivating enzyme, and concentrating to obtain an extract after enzymolysis;
step 3, adsorption;
adding composite adsorbent with mass being 20% -30% of the mass of the extract into the extract after enzymolysis, adsorbing icariin, heating to 55-65 ℃, stirring, filtering, and collecting precipitate and mother liquor; the composite adsorbent consists of active carbon, bentonite and chitosan;
step 4, resolving;
adding isopropanol into the precipitate obtained in step 3, heating for dissolving, filtering, concentrating the filtrate, cooling for crystallizing, adding acetone, heating, rinsing, and drying to obtain icariine with content of more than 98%;
adsorbing the mother liquor obtained in the step 3 by using cationic resin, washing the mother liquor to be neutral by using water, and then eluting the mother liquor by using ammonia alcohol; adding clarifier into the adsorption effluent for clarification, adding protease for enzymolysis, filtering with ultrafiltration membrane, and collecting the concentrated solution to obtain herba Epimedii polysaccharide; concentrating the ammonia alcohol eluent to obtain epimedium total alkaloids.
Further, in the step 1, epimedium is epimedium wushanense; the organic solvent is acetone with pH of 8-8.5 and concentration of 70-90%, and the amount of acetone is 0.5-1.5% (ml/g) of herba Epimedii raw material.
Further, in the step 1, the pressure of the supercritical carbon dioxide extraction method is 38-45MPa, the extraction temperature is 45-50 ℃, the extraction time is 3-4h, the pressure of the first-stage separation kettle is 8-10MPa, the temperature is 40-45 ℃, the pressure of the second-stage separation kettle is 6-8MPa, and the temperature is 35-40 ℃.
Further, in order to increase the content of icariine in the raw materials to the maximum, in step 2.1, the alkaline water is ammonia water or ammonium hydroxide with the mass fraction of 5-8%; the mass of the active carbon is 0.4-0.6% of the mass of the extraction residue; concentrating the filtrate to 1.02-1.03;
in the step 2.2, the mass ratio of the calcium chloride to the magnesium sulfate is 2.8:1, and the addition amount is 0.01-0.02% of the mass of the extraction slag in the step 1; concentrating the specific gravity to 1.05-1.08 after enzyme deactivation;
the adding amount of the rhamnosidase is 7-9% of the mass of the extraction residue in the step 1, and the enzyme activity is 1600 u/ml; the addition amount of xylosidase is 0.5-1% of the extraction residue in step 1, and the enzyme activity is 500 u/ml.
Further, the adding amount of the rhamnosidase is 8% of the mass of the extraction residue in the step 1; the addition amount of xylosidase is 0.6% of the extraction residue in step 1, and the addition amount of calcium chloride and magnesium sulfate is 0.015% of the extraction residue in step 1.
Further, in order to fully adsorb icariin, in step 3, the composite adsorbent comprises 70-80% of activated carbon, 5-10% of bentonite and 15-20% of chitosan.
Furthermore, the composite adsorbent comprises 75% of activated carbon, 9% of bentonite and 16% of chitosan.
Further, in the step 4, the concentration of the isopropanol is 60-80%, and the dosage of the isopropanol is 8-10 times of the mass of the precipitate obtained in the step 3.
Further, in order to improve the recovery rate of the effective components, the dosage of the protease in the step 5 is 0.2-0.5% of the mass of the extraction residue in the step 1, the enzymolysis temperature is 40-50 ℃, the enzymolysis time is 1.5-2.5h, the enzymolysis pH is 6-7, and the cut-off molecular weight of the ultrafiltration membrane is 20-50 KD; the ammonia alcohol eluent is 50% ethanol with 5% ammonia water mass concentration, and the dosage is 3 BV.
The invention has the beneficial effects that:
1. the invention adopts a complex enzyme enzymolysis process to convert the high-grade glucoside in the epimedium into the icariine as much as possible, thereby increasing the content of the icariine in the raw materials to the maximum extent, reducing the cost, improving the extraction rate and leading the recovery rate of the effective components to reach more than 90 percent. And by adding a mixed reagent of calcium chloride and magnesium sulfate, the enzymolysis effect is greatly improved, the use amount of enzyme is reduced, a good foundation is laid for subsequent separation and purification, and the yield is further improved.
2. The invention abandons the traditional process of macroporous adsorption resin, develops a new way, adopts the composite adsorbent to adsorb the icariine, has good selectivity, can be well separated from water-soluble components, reduces the use of organic solvent and the treatment of acid and alkali, reduces the environmental pollution, ensures that the whole process is simple and easy to operate, avoids the dead adsorption of the icariine in column separation, and improves the yield. Meanwhile, the water-soluble components are adsorbed by adopting cationic resin, so that epimedium polysaccharide and alkaloid can be well separated, and the recovery rate of the effective components is improved.
3. The invention adopts the supercritical carbon dioxide extraction technology to separate the chlorophyll in the epimedium raw material, has mild conditions and avoids the degradation of the chlorophyll. Not only can obtain chlorophyll additional products, but also provides a good foundation for the separation, purification and identification of icariin.
4. The invention adopts the combined use of a plurality of technologies of supercritical extraction, enzymolysis, adsorption, desorption, ionic resin purification and membrane filtration, develops the active ingredients in the epimedium raw materials to the maximum extent, obtains chlorophyll, epimedium polysaccharide and alkaloid except the icariin, greatly reduces the production cost of a single product, and greatly improves the production efficiency.
Drawings
FIG. 1 is a process flow diagram of the method of the present invention;
FIG. 2 is an HPLC chromatogram of Epimedium wushanense raw material;
FIG. 3 is a diagram showing the enzymolysis effect of the mixed reagent of calcium chloride and magnesium sulfate;
FIG. 4 is a diagram showing the effect of enzymolysis by adding a mixed reagent of calcium chloride and magnesium sulfate;
FIG. 5 effect of temperature on enzyme activity;
FIG. 6 is a HPLC chromatogram of the mother liquor after adsorption by the composite adsorbent;
FIG. 7 is an HPLC chromatogram of icariin in example one of the present invention;
FIG. 8 is an HPLC chromatogram of icariin in example II of the present invention;
FIG. 9 is an HPLC chromatogram of icariin in example III of the present invention;
FIG. 10 is an HPLC chromatogram of icariin in example IV of the present invention;
FIG. 11 is an HPLC chromatogram of icariin in example V of the present invention;
FIG. 12 is an HPLC chromatogram of icariin in example six of the present invention.
Detailed Description
The invention is further described with reference to the following figures and specific embodiments.
Referring to fig. 1, the preparation method mainly comprises the processes of carbon dioxide supercritical extraction, enzymolysis, adsorption, resolution and preparation of polysaccharide and alkaloid, wherein in the process of carbon dioxide supercritical extraction, a weak alkaline organic solvent is used as an entrainer, and the separated extraction liquid is chlorophyll; calcium chloride and magnesium sulfate are added in the extraction and enzymolysis processes, so that the content of icariine in the raw materials is increased to the maximum extent, the cost for preparing the icariine is reduced, and the extraction yield is improved. And through experimental verification, specifically comparing fig. 3 with fig. 4, fig. 3 shows the enzymolysis effect of the mixed reagent of calcium chloride and magnesium sulfate which is not added, the content of icariine after enzymolysis is 14.52%, while fig. 4 shows the enzymolysis effect of the mixed reagent of calcium chloride and magnesium sulfate which is added, the content of icariine after enzymolysis is 27.92%, thus obtaining the icariine, and the mixed reagent of calcium chloride and magnesium sulfate which is added can greatly improve the enzymolysis effect, reduce the usage amount of enzyme, and lay a good foundation for subsequent separation and purification. And the influence of temperature on the enzyme activity was examined, as shown in FIG. 5, it can be seen that the enzyme activity was high at 45-55 ℃ and thus the following examples performed the enzymatic hydrolysis process in this temperature range. In the adsorption and desorption process, the icariine is adsorbed by the composite adsorbent, as shown in fig. 6, after the icariine is adsorbed by the composite adsorbent, the icariine in the mother liquor has no content basically, and the icariine is completely adsorbed without loss. The use of organic solvents and the treatment of acid and alkali are reduced, the environmental pollution is reduced, the whole process is simple and easy to operate, the epimedium polysaccharide and the alkaloid except the icariine can be obtained by simple operation, the production cost of a single product is greatly reduced, and the production efficiency is greatly improved.
Example one
The embodiment is realized by the following processes:
taking 100Kg of Wushan epimedium raw material, pulverizing to 80 mesh, and its HPLC spectrogram is shown in figure 2, which shows that the icariine content is 0.3%. Taking acetone with the mass of 1 percent of the mass of the raw material, the pH value of 8 and the concentration of 80 percent as an entrainer, adopting carbon dioxide supercritical extraction, wherein the extraction pressure is 38MPa, the extraction temperature is 45 ℃, the extraction time is 3h, the pressure of a separation kettle of a first stage is 8MPa, the temperature is 45 ℃, the pressure of a separation kettle of a second stage is 6MPa, the temperature is 40 ℃, and separating to obtain chlorophyll and extraction residues. Chlorophyll can be prepared into sodium copper chlorophyllin by conventional process.
The extraction residue is boiled for 1h by adopting ammonia water with the mass fraction of 5 percent, cooled at normal temperature, adding active carbon with the mass of 0.5% of the extraction residue, uniformly mixing, extracting for 2 times by adopting 40% ethanol with the mass of 5 times of the mass of the extraction residue under normal temperature stirring, each time for 1.5h, filtering, combining the two filtrates, concentrating the combined filtrate until the specific gravity is 1.02, adding a mixed reagent of calcium chloride and magnesium sulfate with the mass of 0.02% of the extraction residue (wherein the mass ratio of the calcium chloride to the magnesium sulfate is 2.8:1), stirring for 30min, filtering, adding rhamnosidase with the mass of 7% of the extraction residue and xylosidase with the mass of 0.5% into the filtrate for enzymolysis for 3h at 50 ℃, inactivating enzyme, concentrating until the specific gravity is 1.05, obtaining an extract after enzymolysis, wherein the content of icariine in the raw material is 1.71% after enzymolysis, and is increased by 4.7 times compared with the original content of the icariine in the raw material which is 0.3%; then adding composite adsorbent (wherein activated carbon accounts for 70%, bentonite accounts for 10%, and chitosan accounts for 20%) 20% of the mass of the extract, stirring at 55 deg.C for 1h, filtering, and collecting precipitate and mother liquor respectively.
Dissolving the precipitate with 8 times of 80% isopropanol by heating, filtering, concentrating the filtrate, cooling, crystallizing, adding hot acetone, rinsing, and drying to obtain 1.58Kg solid, and detecting by HPLC (high performance liquid chromatography) as shown in FIG. 7, wherein the icariine content is 98.3%. The yield is 1.58%, which is equivalent to the content of the icariine in the raw material being 1.55%, and compared with the content of the icariine in the raw material after enzymolysis being 1.71%, the recovery rate of the effective components is 90.6%.
The mother liquor is absorbed by 001 × 7 cation resin (the absorption amount is 12g crude drug/ml resin), washed to neutrality by water, and then eluted by 3BV ammonia alcohol (50% ethanol with 5% ammonia water mass concentration). Clarifying the adsorption effluent with clarifier, adding protease 45 deg.C (pH6-7) with a mass of 0.2% of the extraction residue, performing enzymolysis for 2 hr, filtering with ultrafiltration membrane with cut-off molecular weight of 20KD, and collecting the concentrated solution to obtain epimedium polysaccharide 8.35Kg with content of 82.6%. Concentrating the ammonia alcohol eluate to obtain herba Epimedii total alkaloids 0.82Kg with content of 67.3%.
Example two
The embodiment is realized by the following processes:
taking 100Kg of Wushan epimedium raw material (the icariin content is 0.3%), crushing the Wushan epimedium raw material into 80 meshes, taking acetone with the mass of 0.5 percent, the pH value of 8.5 and the concentration of 70 percent as an entrainer, adopting carbon dioxide supercritical extraction, extracting at the temperature of 50 ℃ and the extraction time of 4h under the pressure of 45MPa in a separation kettle of the I stage, at the temperature of 40 ℃ and 8MPa in a separation kettle of the II stage, at the temperature of 35 ℃, and separating to obtain chlorophyll and extraction residues. Chlorophyll can be prepared into sodium copper chlorophyllin by conventional process.
The obtained extraction residue is boiled by 8% ammonia water for 1h, cooled at normal temperature, added with 0.4% active carbon and mixed evenly, 40% ethanol with the mass 5 times of that of the extraction residue is adopted to stir and extract for 2 times at normal temperature, 1.5h each time, filtered, the two filtrates are combined, the combined filtrate is concentrated to the specific gravity of 1.03, a mixed reagent of calcium chloride and magnesium sulfate with the mass 0.01% of that of the extraction residue is added (wherein, the mass ratio of the calcium chloride to the magnesium sulfate is 2.8:1), stirred for 30min and filtered, rhamnosidase and 1% xylosidase with the mass 9% of that of the extraction residue are added into the filtrate, the enzymolysis is carried out for 3h at 45 ℃, the enzyme is deactivated, the concentration is carried out to the specific gravity of 1.06, an extract after the enzymolysis is obtained, the content of the raw material icariin the amount of 1.96% after the enzymolysis is increased by 5.5 times compared with the content of the original icariin the raw material of 0.3%; then adding composite adsorbent (wherein the mass of the activated carbon accounts for 80%, the mass of the bentonite accounts for 5%, and the mass of the chitosan accounts for 15%) with the mass of 30% of the mass of the extract, stirring for 1h at 65 ℃, filtering, and respectively collecting precipitate and mother liquor.
Dissolving the precipitate with 80% isopropanol 10 times the mass of the precipitate, filtering, concentrating the filtrate, cooling, crystallizing, adding hot acetone, rinsing, and drying to obtain 1.82Kg solid, and detecting by HPLC (high performance liquid chromatography), as shown in FIG. 8, wherein the icariine content is 98.2%. The yield is 1.82%, which is equivalent to 1.79% of the content of the icariine in the raw materials, and the recovery rate of the effective components is 91.3% compared with the 1.96% of the content of the icariine in the raw materials after enzymolysis.
The mother liquor is absorbed by 001 × 7 cation resin (the absorption amount is 12g crude drug/ml resin), washed to neutrality by water, and then eluted by 3BV ammonia alcohol (50% ethanol with 5% ammonia water mass concentration). Clarifying the adsorption effluent with clarifier, adding protease 40 deg.C (pH6-7) with a mass of 0.5% of the extraction residue for enzymolysis for 2.5 hr, filtering with ultrafiltration membrane with a molecular weight cut-off of 50KD, and collecting the concentrated solution to obtain herba Epimedii polysaccharide 7.95Kg with a content of 85.2%. Concentrating the ammonia alcohol eluate to obtain herba Epimedii total alkaloids 0.84Kg with content of 66.0%.
EXAMPLE III
100Kg of Wushan epimedium raw material (with icariin content of 0.3%) is taken and crushed into 80 meshes, acetone with the mass of 1.5%, pH 8 and concentration of 90% of the raw material is taken as an entrainer, carbon dioxide supercritical extraction is adopted, the extraction pressure is 40MPa, the extraction temperature is 48 ℃, the extraction time is 4h, the pressure of a separation kettle for I stage is 9MPa, the temperature is 42 ℃, the pressure of a separation kettle for II stage is 7MPa, the temperature is 38 ℃, and chlorophyll and extraction residues are obtained after separation. Chlorophyll can be prepared into sodium copper chlorophyllin by conventional process.
Cooking the obtained extraction residue for 1 hour by adopting 6% ammonium hydroxide by mass fraction, cooling at normal temperature, then adding 0.6% by mass of active carbon into the extraction residue, uniformly mixing, stirring and extracting for 2 times at normal temperature by adopting 40% ethanol with the mass being 5 times of the mass of the extraction residue, 1.5 hours each time, filtering, combining the two filtrates, concentrating the combined filtrate to the specific gravity of 1.02, adding 0.012% by mass of a mixed reagent of calcium chloride and magnesium sulfate (mass ratio is 2.8:1) into the extraction residue, stirring for 30 minutes, filtering, adding 8% by mass of rhamnosidase and 0.9% by mass of xylosidase into the filtrate, performing enzymolysis for 3 hours at 55 ℃, inactivating enzymes, and concentrating to the specific gravity of 1.08 to obtain an extract after enzymolysis; after enzymolysis, the content of icariine in the raw material is 1.88 percent, which is 5.3 times higher than the original content of icariine in the raw material of 0.3 percent; then adding composite adsorbent (72% of active carbon, 8% of bentonite and 20% of chitosan) with mass of 25% of the extract mass, stirring at 60 deg.C for 1h, filtering, and collecting precipitate and mother liquor respectively.
Dissolving the precipitate with 80% isopropanol 9 times the mass of the precipitate by heating, filtering, concentrating the filtrate, cooling for crystallization, rinsing with hot acetone, and drying to obtain 1.75Kg of solid, which is detected by HPLC (high performance liquid chromatography), as shown in FIG. 9, and has icariine content of 98.5%. The yield is 1.75 percent, which is equivalent to the content of the icariine in the raw material being 1.72 percent, and the recovery rate of the effective components is 91.5 percent compared with the icariine content in the raw material after enzymolysis which is 1.88 percent.
The mother liquor is absorbed by 001 × 7 cation resin (the absorption amount is 12g crude drug/ml resin), washed to neutrality by water, and then eluted by 3BV ammonia alcohol (50% ethanol with 5% ammonia water mass concentration). Clarifying the adsorption effluent with clarifier, adding protease 0.3% of the extraction residue by mass at 50 deg.C (pH6-7) for enzymolysis for 2h, filtering with ultrafiltration membrane with cut-off molecular weight of 30KD, and collecting the concentrated solution to obtain epimedium polysaccharide 8.34Kg with content of 80.9%. Concentrating the ammonia alcohol eluate to obtain herba Epimedii total alkaloids 0.80Kg with content of 68.3%.
Example four
100Kg of Wushan epimedium raw material (with icariin content of 0.3%) is taken and crushed into 80 meshes, acetone with the mass of 1%, pH 8 and concentration of 80% of the raw material is taken as an entrainer, carbon dioxide supercritical extraction is adopted, the extraction pressure is 40MPa, the extraction temperature is 46 ℃, the extraction time is 3h, the pressure of a separation kettle for I stage is 9MPa, the temperature is 43 ℃, the pressure of a separation kettle for II stage is 7MPa, the temperature is 36 ℃, and chlorophyll and extraction residues are obtained after separation. Chlorophyll can be prepared into sodium copper chlorophyllin by conventional process.
Cooking the obtained extraction residue for 1 hour by using 8% ammonium hydroxide by mass fraction, cooling at normal temperature, adding 0.5% by mass of active carbon into the extraction residue, uniformly mixing, stirring and extracting for 2 times at normal temperature by using 40% ethanol with the mass being 5 times of the mass of the extraction residue, 1.5 hours each time, filtering, combining the two filtrates, concentrating the combined filtrate to the specific gravity of 1.02, adding 0.015% by mass of a mixed reagent of calcium chloride and magnesium sulfate (mass ratio is 2.8:1), stirring for 30 minutes, filtering, adding rhamnosidase with the mass being 8% of the mass of the extraction residue and 0.6% of xylosidase into the filtrate, carrying out enzymolysis for 3 hours at 50 ℃, inactivating the enzymes, concentrating to the specific gravity of 1.05 to obtain an extract after enzymolysis, wherein the content of icariine in the raw material after the enzymolysis is 1.94%, and is increased by 5.5 times compared with the original content of the icariine in the raw material which is 0.3%; then adding composite adsorbent (wherein activated carbon accounts for 75%, bentonite accounts for 9%, and chitosan accounts for 16%) with liquid volume of 25%, stirring at 60 deg.C for 1 hr, filtering, and collecting precipitate and mother liquor respectively. Dissolving the precipitate with 10 times of 80% isopropanol under heating, filtering, concentrating the filtrate, cooling, crystallizing, rinsing with hot acetone, and drying to obtain 1.83Kg solid, which is detected by HPLC (high performance liquid chromatography) as shown in FIG. 10, and contains icariine 98.7%. The yield is 1.8 and 3 percent, the content of the icariine in the raw material is 1.83 percent, and the recovery rate of the effective components is 93.3 percent compared with the icariine content of the raw material after enzymolysis which is 1.94 percent.
The mother liquor is absorbed by 001 × 7 cation resin (the absorption amount is 12g crude drug/ml resin), washed to neutrality by water, and then eluted by 3BV ammonia alcohol (50% ethanol with 5% ammonia water mass concentration). Clarifying the adsorption effluent with clarifier, adding protease 45 deg.C (pH6-7) with a mass of 0.4% of the extraction residue, performing enzymolysis for 2 hr, filtering with ultrafiltration membrane with cut-off molecular weight of 40KD, and collecting the concentrated solution to obtain epimedium polysaccharide 8.04Kg with content of 85.2%. Concentrating the ammonia alcohol eluate to obtain herba Epimedii total alkaloids 0.82Kg with content of 70.2%.
EXAMPLE five
100Kg of Wushan epimedium raw material (with icariin content of 0.3%) is taken and crushed into 80 meshes, acetone with the mass of 1%, pH 8 and concentration of 80% of the raw material is taken as an entrainer, carbon dioxide supercritical extraction is adopted, the extraction pressure is 42MPa, the extraction temperature is 45 ℃, the extraction time is 3h, the separation kettle pressure of I stage is 10MPa, the extraction temperature is 40 ℃, the separation kettle pressure of II stage is 8MPa, the extraction temperature is 35 ℃, and chlorophyll and extraction residues are obtained after separation. Chlorophyll can be prepared into sodium copper chlorophyllin by conventional process.
The obtained extraction residue is boiled by 7% ammonia water for 1 hour, cooled at normal temperature, added with 0.5% active carbon by mass of the extraction residue and uniformly mixed, 40% ethanol with the mass 5 times of the mass of the extraction residue is adopted for stirring and extracting for 2 times at normal temperature, each time lasts for 1.5 hours, the two filtrates are combined, the combined filtrate is concentrated to the specific gravity of 1.02, a mixed reagent of calcium chloride and magnesium sulfate with the mass of 0.018% of the mass of the extraction residue is added (wherein, the mass ratio of the calcium chloride to the magnesium sulfate is 2.8:1), the mixture is stirred for 30min and filtered, rhamnosidase with the mass of 9% of the mass of the extraction residue and xylosidase with the mass of 0.7% are added into the filtrate for enzymolysis for 3 hours at 50 ℃, the enzymes are inactivated, the mixture is concentrated to the specific gravity of 1.05, an extract after enzymolysis is obtained, the content of the raw material is reduced to 1.89% after the enzymolysis, and is increased by 5.3 times compared with the original content of; then adding composite adsorbent (76% of active carbon, 6% of bentonite and 18% of chitosan) with liquid volume of 28%, stirring at 60 deg.C for 1h, filtering, and collecting precipitate and mother liquor respectively.
Dissolving the precipitate with 10 times of 80% isopropanol by heating, filtering, concentrating the filtrate, cooling for crystallization, rinsing with hot acetone, and drying to obtain 1.77Kg solid, which is detected by HPLC (high performance liquid chromatography), as shown in FIG. 11, and has icariine content of 98.3%. The yield is 1.77%, which is equivalent to 1.57% of the icariine content in the raw material, and the recovery rate of the effective components is 92.1% compared with the 1.89% of the icariine content in the raw material after enzymolysis.
The mother liquor is absorbed by 001 × 7 cation resin (the absorption amount is 12g crude drug/ml resin), washed to neutrality by water, and then eluted by 3BV ammonia alcohol (50% ethanol with 5% ammonia water mass concentration). Clarifying the adsorption effluent with clarifier, adding protease 50 deg.C (pH6-7) with a mass of 0.5% of the extract residue, performing enzymolysis for 1.5 hr, filtering with ultrafiltration membrane with cut-off molecular weight of 20KD, and collecting the concentrated solution to obtain epimedium polysaccharide 8.29Kg with a content of 80.4%. Concentrating the ammonia alcohol eluate to obtain herba Epimedii total alkaloids 0.80Kg with content of 68.0%.
EXAMPLE six
100Kg of Wushan epimedium raw material (with icariin content of 0.3%) is taken and crushed into 80 meshes, acetone with the mass of 1%, pH 8 and concentration of 80% of the raw material is taken as an entrainer, carbon dioxide supercritical extraction is adopted, the extraction pressure is 39MPa, the extraction temperature is 49 ℃, the extraction time is 4h, the pressure of a separation kettle for I stage is 8MPa, the temperature is 43 ℃, the pressure of a separation kettle for II stage is 7MPa, the temperature is 39 ℃, and chlorophyll and extraction residues are obtained after separation. Chlorophyll can be prepared into sodium copper chlorophyllin by conventional process.
The obtained extraction residue is steamed and boiled for 1 hour by adopting 5% ammonium hydroxide by mass fraction, cooled at normal temperature, added with active carbon with the mass of 0.5% of the extraction residue and uniformly mixed, stirred and extracted for 2 times at normal temperature by adopting 40% ethanol with the mass of 5 times of the mass of the extraction residue, and each time lasts for 1.5 hours, filtered, the two filtrates are combined, the combined filtrate is concentrated to the specific gravity of 1.02, added with a mixed reagent (the mass ratio is 2.8:1) of calcium chloride and magnesium sulfate with the mass of 0.01% of the mass of the extraction residue and stirred for 30 minutes, filtered, added with rhamnosidase and xylosidase with the mass of 7% of the raw materials for 3 hours at 50 ℃, inactivated and concentrated to the specific gravity of 1.05 to obtain an extract after enzymolysis, the content of the raw materials is reduced to 1.79% after the enzymolysis, and is increased by 5 times compared with the original content of the icariine in the raw materials of 0.3%; then adding composite adsorbent (containing 79% of active carbon, 7% of bentonite and 14% of chitosan) with mass of 28% of the extract, stirring at 60 deg.C for 1h, filtering, and collecting precipitate and mother liquor respectively.
Dissolving the precipitate with 8 times of 80% isopropanol by heating, filtering, concentrating the filtrate, cooling for crystallization, rinsing with hot acetone, and drying to obtain 1.65Kg solid, which is detected by HPLC (high performance liquid chromatography) as shown in FIG. 12, wherein the icariine content is 98.5%. The yield is 1.65%, which is equivalent to the content of the icariine in the raw material being 1.63%, and compared with the content of the icariine in the raw material after enzymolysis being 1.79%, the recovery rate of the effective components is 91.1%.
The mother liquor is absorbed by 001 × 7 cation resin (the absorption amount is 12g crude drug/ml resin), washed to neutrality by water, and then eluted by 3BV ammonia alcohol (50% ethanol with 5% ammonia water mass concentration). Clarifying the adsorption effluent with clarifier, adding protease 45 deg.C (pH6-7) with a mass of 0.2% of the extracted residue, performing enzymolysis for 2 hr, filtering with ultrafiltration membrane with a molecular weight cutoff of 50KD, and collecting the concentrated solution to obtain epimedium polysaccharide 8.03Kg with a content of 82.4%. Concentrating the ammonia alcohol eluate to obtain herba Epimedii total alkaloids 0.83Kg with content of 68.6%.
Claims (9)
1. A preparation method of icariin is characterized by comprising the following steps:
step 1, supercritical extraction;
pulverizing herba Epimedii raw material, extracting herba Epimedii raw material with weakly alkaline organic solvent as entrainer by supercritical carbon dioxide extraction method to obtain extractive solution and extraction residue, wherein the extractive solution is chlorophyll;
step 2, extracting and performing enzymolysis;
step 2.1, cooking the extraction residue obtained in the step 1 with alkaline water, then adding activated carbon, mixing uniformly, adding ethanol, stirring and extracting at normal temperature, filtering, and concentrating the filtrate;
step 2.2, adding a mixed reagent of calcium chloride and magnesium sulfate into the filtrate obtained in the step 2.1, stirring, filtering, adding rhamnosidase and xylosidase into the filtrate for enzymolysis, wherein the enzymolysis temperature is 45-55 ℃, inactivating enzyme, and concentrating to obtain an extract after enzymolysis;
step 3, adsorption;
adding composite adsorbent with mass being 20% -30% of the mass of the extract into the extract after enzymolysis, adsorbing icariin, heating to 55-65 ℃, stirring, filtering, and collecting precipitate and mother liquor; the composite adsorbent consists of active carbon, bentonite and chitosan;
step 4, resolving;
adding isopropanol into the precipitate obtained in step 3, heating for dissolving, filtering, concentrating the filtrate, cooling for crystallizing, rinsing with hot acetone, and drying to obtain icariine with content of more than 98%;
step 5, preparing polysaccharide and alkaloid;
adsorbing the mother liquor obtained in the step 3 by using cationic resin, washing the mother liquor to be neutral by using water, and then eluting the mother liquor by using ammonia alcohol; adding clarifier into the adsorption effluent for clarification, adding protease for enzymolysis, filtering with ultrafiltration membrane, and collecting the concentrated solution to obtain herba Epimedii polysaccharide; concentrating the ammonia alcohol eluent to obtain epimedium total alkaloids.
2. The method for preparing icariin according to claim 1, wherein:
in the step 1, epimedium is epimedium wushanense; the organic solvent is acetone with pH of 8-8.5 and concentration of 70-90%, and the amount of acetone is 0.5-1.5% (ml/g) of herba Epimedii raw material.
3. The method for preparing icariin according to claim 2, wherein: in the step 1, the pressure of the supercritical carbon dioxide extraction method is 38-45MPa, the extraction temperature is 45-50 ℃, the extraction time is 3-4h, the pressure of a first-stage separation kettle is 8-10MPa, the temperature is 40-45 ℃, the pressure of a second-stage separation kettle is 6-8MPa, and the temperature is 35-40 ℃.
4. The method for preparing icariin according to claim 3, wherein: in the step 2.1, the alkaline water is ammonia water or ammonium hydroxide with the mass fraction of 5-8%; the mass of the active carbon is 0.4-0.6% of the mass of the extraction residue; concentrating the filtrate to 1.02-1.03;
in the step 2.2, the mass ratio of the calcium chloride to the magnesium sulfate is 2.8:1, and the addition amount is 0.01-0.02% of the mass of the extraction slag in the step 1; concentrating the specific gravity to 1.05-1.08 after enzyme deactivation;
the adding amount of the rhamnosidase is 7-9% of the mass of the extraction residue in the step 1, and the enzyme activity is 1600 u/ml; the addition amount of xylosidase is 0.5-1% of the extraction residue in step 1, and the enzyme activity is 500 u/ml.
5. The method for preparing icariin according to claim 4, wherein: the addition amount of the rhamnosidase is 8% of the mass of the extraction residue in the step 1; the addition amount of xylosidase is 0.6 percent of the mass of the extraction residue in the step 1; the addition amount of calcium chloride and magnesium sulfate is 0.015 percent of the mass of the extraction slag in the step 1.
6. The method for preparing icariin according to claim 4, wherein: in the step 3, the composite adsorbent comprises 70-80% of active carbon, 5-10% of bentonite and 15-20% of chitosan.
7. The method for preparing icariin according to claim 6, wherein: the composite adsorbent comprises 75% of activated carbon, 9% of bentonite and 16% of chitosan.
8. The method for preparing icariin according to claim 6, wherein: in the step 4, the concentration of the isopropanol is 60-80%, and the dosage of the isopropanol is 8-10 times of the mass of the precipitate obtained in the step 3.
9. The method for preparing icariin according to claim 6, wherein: in the step 5, the dosage of the protease is 0.2-0.5% of the mass of the extraction residue in the step 1, the enzymolysis temperature is 40-50 ℃, the enzymolysis time is 1.5-2.5h, the enzymolysis pH is 6-7, and the cut-off molecular weight of an ultrafiltration membrane is 20-50 KD; the ammonia alcohol eluent is 50% ethanol with 5% ammonia water mass concentration, and the dosage is 3 BV.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1535976A (en) * | 2003-04-11 | 2004-10-13 | 中国科学院过程工程研究所 | Method for extracting icariine from epimedium plant |
| CN1724005A (en) * | 2005-07-18 | 2006-01-25 | 天津大学 | Method for extracting volatile oil and liposoluble alkaloid from plumula nelumbini |
| WO2008138243A1 (en) * | 2007-05-09 | 2008-11-20 | Beijing Shenogen Biomedical Co., Ltd | A preparation method of icaritin |
| CN101845067A (en) * | 2010-05-21 | 2010-09-29 | 甘肃亚兰特种药材饮片生产有限公司 | Preparation method of high-content icariin |
| CN105461766A (en) * | 2015-12-18 | 2016-04-06 | 陕西嘉禾生物科技股份有限公司 | Method for extracting salicin from white willow bark |
| CN109295121A (en) * | 2018-11-05 | 2019-02-01 | 重庆大学 | A kind of method that enzymatic isolation method prepares icariin |
-
2020
- 2020-05-08 CN CN202010383284.2A patent/CN111560409B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1535976A (en) * | 2003-04-11 | 2004-10-13 | 中国科学院过程工程研究所 | Method for extracting icariine from epimedium plant |
| CN1724005A (en) * | 2005-07-18 | 2006-01-25 | 天津大学 | Method for extracting volatile oil and liposoluble alkaloid from plumula nelumbini |
| WO2008138243A1 (en) * | 2007-05-09 | 2008-11-20 | Beijing Shenogen Biomedical Co., Ltd | A preparation method of icaritin |
| CN101845067A (en) * | 2010-05-21 | 2010-09-29 | 甘肃亚兰特种药材饮片生产有限公司 | Preparation method of high-content icariin |
| CN105461766A (en) * | 2015-12-18 | 2016-04-06 | 陕西嘉禾生物科技股份有限公司 | Method for extracting salicin from white willow bark |
| CN109295121A (en) * | 2018-11-05 | 2019-02-01 | 重庆大学 | A kind of method that enzymatic isolation method prepares icariin |
Non-Patent Citations (4)
| Title |
|---|
| YUNBIN LYU等: "Efficient bioconversion of epimedin C to icariin by a glycosidase from Aspergillus nidulans", 《BIORESOURCE TECHNOLOGY》 * |
| 娄方明等: "淫羊藿中淫羊藿苷分离纯化工艺研究", 《安徽农业科学》 * |
| 肖利萍 等: "改性膨润土吸附强化混凝处理微污染水源水", 《非金属矿》 * |
| 陈乃松等: "酶制剂体外酶解豆粕中抗营养因子的研究", 《大豆科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114249777A (en) * | 2021-11-19 | 2022-03-29 | 安徽金源药业有限公司 | Method for extracting icariin |
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