CN113980065A - Method for extracting icariin from epimedium herb - Google Patents
Method for extracting icariin from epimedium herb Download PDFInfo
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- CN113980065A CN113980065A CN202111317176.6A CN202111317176A CN113980065A CN 113980065 A CN113980065 A CN 113980065A CN 202111317176 A CN202111317176 A CN 202111317176A CN 113980065 A CN113980065 A CN 113980065A
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- icariin
- ethyl acetate
- epimedin
- filtering
- crude extract
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- 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
- C07H1/08—Separation; Purification from natural products
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- 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
Abstract
The invention provides an extraction method of icariin, which comprises the steps of soaking epimedium herb powder in 1.4-1.5 times of sodium carbonate aqueous solution with the pH value of 10-11 by weight for about half an hour, then introducing steam to stir for about 1 hour, and carrying out hot filtration; introducing water vapor into the solid slag for about 1 hour, stirring, carrying out hot filtration, and repeating the steps once; mixing filtrates, cooling in ice bath, adjusting pH to 3.5-4.2, standing for precipitation, filtering, collecting precipitate, and drying at low temperature; mixing the dried crude extract with C18 bonded silica gel, performing column chromatography, detecting with HPLC during elution, retaining the chromatogram of the reference for 10.5 min, and collecting the same components; the eluent is recovered by distillation and recrystallization is carried out twice. High-purity icariin and other flavonoids such as epimedin A, epimedin B, epimedin C and the like can be obtained, wherein the icariin accounts for 0.8 weight part, the content is over 98.4 percent, and the extraction rate is over 97.5 percent.
Description
Technical Field
The invention belongs to the technical field of extraction of plant effective components, and relates to a method for extracting icariin from epimedium herb.
Background
Icariin (also called icariin) has the activities of improving the organism immunity by regulating the balance of T cells, promoting the generation and activation of osteoblasts, inhibiting osteoporosis, protecting cardiac muscle cells, increasing the number of the cardiac muscle cells, improving endothelial dysfunction, repairing cardiovascular and cerebrovascular functions, regulating endocrine, resisting bacteria, diminishing inflammation, resisting oxidation, aging and tumors, enhancing the learning and memory ability, improving reproductive function and the like, and can also treat diseases such as Alzheimer's disease, Parkinson, cerebral ischemia, atherosclerosis, rheumatism and the like. Icariin is the main active ingredient of epimedium, and the epimedium is recorded in 2020 edition of the ministerial part of the Chinese pharmacopoeia to have functions and main indications: tonifying kidney yang, strengthening tendons and bones, dispelling wind-damp, being used for kidney yang deficiency, impotence and spermatorrhea, flaccidity of bones and muscles, rheumatic arthralgia and numbness and contracture, and indicating that icariin is a marker for evaluating the quality of epimedium medicinal materials and is also a basis for evaluating the quality and quality of extracts in the deep processing production of epimedium. The content of icariin in the whole herba epimedii is lower, usually 0.2-3.0%, and the content of icariin can be different according to different harvesting seasons, varieties, production places, application positions, processing modes and the like.
The existing icariin is prepared from epimedium medicinal materials serving as raw materials, ethanol with different concentrations serving as solvents is extracted by heating reflux, ultrasound, microwave, leakage and other methods to obtain crude extracts of epimedium, and the crude extracts are extracted, separated and refined to obtain high-purity icariin from the epimedium medicinal materials. The epimedium herb contains icariin,The content of epimedin A, epimedin B, epimedin C and other flavonoids is lower, and referring to the method for measuring the content of icariin in epimedium medicinal materials in the 2020 edition of the Chinese pharmacopoeia, the content of icariin in epimedium medicinal materials requires that the epimedin (C) is contained33H40O15) Not less than 0.40%; contains icariin (C) as total flavone33H40O15) Not less than 5.0% ". In the existing process for extracting and preparing high-purity icariin from epimedium medicinal materials, no matter alcohol solution reflux extraction or auxiliary extraction is used, extraction and separation are carried out, and in the process, all steps are complicated, the extraction is incomplete, the loss is large, the content and the extraction rate are unstable, the amount of used organic solvents is large, the types are multiple, the extraction process is complex, the time of the whole process is long, and the cost is high. The existing process for preparing icariin from epimedium medicinal materials is not suitable for extracting low-content raw materials any more.
Disclosure of Invention
The invention aims to provide an icariin extraction method which is high in extraction rate, stable in quality, low in production cost and environment-friendly.
The technical scheme for realizing the purpose is as follows:
a method for extracting icariin from epimedium herb is completed according to the following process steps,
(1) drying herba Epimedii in shade, pulverizing with pulverizer, and sieving with 80 mesh sieve;
(2) weighing 50 parts by weight of herba Epimedii whole plant powder, adding 1.4-1.5 times of sodium carbonate aqueous solution with pH of 10-11 in a distillation retort, soaking for 30-40 min, introducing water vapor, stirring for 50-60 min, and filtering while hot;
(3) introducing water vapor into the solid residue in a distillation tank for 50-60 min, stirring, and filtering;
(4) repeating the step (3) once;
(5) combining the filtrates from the step (2) to the step (4), cooling in an ice bath, adjusting the pH of the filtrate to 3.5-4.2 by using concentrated hydrochloric acid, standing for precipitation, filtering and collecting precipitates;
(6) extracting the mother liquor with ethyl acetate for 2 times, wherein the using amount of the ethyl acetate is 0.6-0.7 time of the volume of the mother liquor each time, recovering the ethyl acetate, combining the solid extracted with the ethyl acetate and the precipitate obtained in the step (5) to obtain a crude extract, and drying at low temperature;
(7) mixing the dried crude extract with C18 bonded silica gel, and mixing the mixture according to the weight ratio: the crude extract and C18 bonded silica gel are 1: 10-12, the wet method is used for column loading, the dry method is used for sample loading, the eluent with the ratio of ethanol to glacial acetic acid to water being 7: 0.5-0.6: 2.4-2.5 is used for elution, HPLC is used for detection, the retention time of the chromatogram of the icariin reference substance is 10.5 min, and the same components are intercepted;
(8) distilling to recover eluent, recrystallizing with ethyl acetate, and recrystallizing with ethanol.
The above steps are completed to obtain high-purity icariin and other flavonoids such as epimedin A, epimedin B, epimedin C and the like, wherein the icariin accounts for 0.8 part by weight, the content of the icariin exceeds 98.4 percent, and the extraction rate of the icariin exceeds 97.5 percent.
The invention has the beneficial effects that:
soaking herba Epimedii powder in sodium carbonate aqueous solution to destroy epidermal cell wall and cell membrane of herba Epimedii, introducing water vapor to soften and loosen cellulose of herba Epimedii, enhancing permeability, promoting full dissolution, and repeatedly extracting for 2 times to make extraction more complete; acidifying the extractive solution with concentrated hydrochloric acid, and extracting the filtrate with ethyl acetate for 2 times to make the extract fully transferred; the crude extract is bonded with a silica gel chromatographic column by using C18, so that the purity and the extraction rate of the icariin are improved, the content of the icariin can be controlled to be 98.4-99.1%, and the extraction rate is 97.5-98.6%. In the whole extraction process, the aqueous solution is mainly used as an extraction medium, so that a large amount of organic solvent is avoided, the extraction and separation processes are simplified, and the used organic solvent and waste water can be recycled.
The invention is different from the prior art in that:
(1) in the prior art, in the extraction process, organic solvents such as methanol, ethanol, acetone and the like or aqueous solutions of the organic solvents with different concentrations are generally used for extraction, other organic solvents such as isopropanol, n-butanol and the like are also used for extraction, and auxiliary extraction technologies such as ultrasound, microwave, high pressure, supercritical carbon dioxide extraction and the like are used for extraction, so that the equipment requirement is high, the working procedures and the operation are complex, and the extraction process is time-consuming and labor-consuming. The method for extracting the cold acid liquid precipitate by using the hot alkali liquid solves the problem of extracting icariin and other flavonoid substances from low-content resources, mainly uses a sodium carbonate aqueous solution as an extraction medium, avoids using a large amount of organic solvent, can recycle waste water, reduces the production cost, and realizes green production.
(2) In the prior art, column chromatography is mostly adopted in the purification and refining processes of icariin, the stationary phase of the column chromatography is commonly used macroporous adsorption resins with different compositions and specifications such as adsorption resin polystyrene, polyacrylamide, polymethacrylate, polyacrylamide and styrene copolymer, methacrylate and styrene copolymer and the like, single eluent or mixed eluent of organic solvents such as methanol, ethanol, acetone, ethyl acetate, n-butyl alcohol and the like is used as a mobile phase, and a plurality of times of column chromatography such as macroporous resin columns, gel columns, silica gel columns and the like are continuously used, so that the working strength is high, the purification time is long, the steps are complicated, and the cost is higher. The invention adopts an economic and simple purification method, uses C18 bonded silica gel chromatographic column, elutes the eluent with the ratio of ethanol, glacial acetic acid and water being 7: 0.5-0.6: 2.4-2.5, and then carries out ethyl acetate recrystallization and ethanol secondary recrystallization to obtain the high-purity icariin.
(3) The icariin extracted by the prior art has low yield and content and unstable product quality. The invention improves the purity and extraction rate of the icariin, the quality of the icariin product is stable and controllable, the content of the icariin can be controlled to be 98.4-99.1%, and the extraction rate is 97.5-98.6%.
Detailed Description
The technical solution of the present invention will be further described with reference to the following examples.
Example one:
drying herba Epimedii in shade, pulverizing with pulverizer, and sieving with 80 mesh sieve; weighing 40 kg of epimedium herb powder in a distillation tank, adding 1.5 times of sodium carbonate aqueous solution with pH of 10 to the weight of the herb powder, enabling the epimedium herb powder to completely and loosely float, soaking for 40 min, introducing water vapor and stirring for 60 min, filtering while hot, introducing water vapor to the solid residue in the distillation tank for 60 min, stirring and extracting for 2 times, filtering, combining filtrates, cooling the filtrate by an ice bath, acidifying with concentrated hydrochloric acid to adjust the pH to 4.2, standing for precipitation, filtering and collecting precipitates; extracting the mother liquor with ethyl acetate at volume ratio of 1: 0.7 for 2 times, recovering ethyl acetate, mixing the solid extracted with ethyl acetate and the precipitate to obtain crude extract, drying at low temperature, and weighing 3.28 kg. Mixing the crude extract with C18 bonded silica gel, and mixing the raw materials in parts by weight: the crude extract and C18 bonded silica gel are 1: 12, wet-process column loading is carried out, dry-process sample loading is carried out, elution is carried out by eluent with the volume ratio of ethanol, glacial acetic acid and water being 7: 0.6: 2.5, HPLC detection is carried out, the same component at 10.5 min of chromatogram retention time is intercepted, eluent is distilled and recovered, ethyl acetate is used for recrystallization, ethanol is used for secondary recrystallization, and other high-purity icariine and other flavonoid substances such as epimedin A, epimedin B, epimedin C and the like can be obtained, wherein the icariine content is 0.64 kg, the epimedin content is 98.4 percent, and the extraction rate is 97.8 percent.
Example two:
drying herba Epimedii in shade, pulverizing with pulverizer, and sieving with 80 mesh sieve; weighing 60 kg of epimedium herb powder in a distillation retort, adding 1.4 times of sodium carbonate aqueous solution with the pH value of 11 in the weight of the herb powder to ensure that the epimedium herb powder is completely and loosely floated, soaking for 30 min, introducing water vapor and stirring for 50 min, filtering while hot, introducing water vapor into the solid slag in the distillation retort for 50 min, stirring and extracting for 2 times, filtering, combining the filtrates, cooling the filtrate by an ice bath, acidifying with concentrated hydrochloric acid to adjust the pH value to 3.5, standing for precipitation, filtering and collecting precipitates; extracting the mother liquor with ethyl acetate at volume ratio of 1: 0.6 for 2 times, recovering ethyl acetate, mixing the solid extracted with ethyl acetate and the precipitate to obtain crude extract, drying at low temperature, and weighing 4.90 kg. Mixing the crude extract with C18 bonded silica gel, and mixing the raw materials in parts by weight: the crude extract and C18 bonded silica gel are 1: 10, the wet method is used for loading the column, the dry method is used for loading, the eluent with the volume ratio of ethanol, glacial acetic acid and water being 7: 0.5: 2.4 is used for elution, HPLC detection is used for detecting, the same component at the position with 10.5 min of chromatogram retention time is intercepted, the eluent is distilled and recovered, ethyl acetate is used for recrystallization, and ethanol is used for secondary recrystallization, and other high-purity icariine and other flavonoid substances such as epimedin A, epimedin B, epimedin C and the like can be obtained, wherein the icariine content is 0.95 kg, the epimedin content is 99.0 percent, and the extraction rate is 97.5 percent.
Example three:
drying herba Epimedii in shade, pulverizing with pulverizer, and sieving with 80 mesh sieve; weighing 80 kg of epimedium herb powder in a distillation retort, adding 1.48 times of sodium carbonate aqueous solution with the pH of 10.5 of the weight of the herb powder to ensure that the epimedium herb powder is completely and loosely floated, soaking for 35 min, introducing water vapor, stirring for 55 min, filtering while hot, introducing the water vapor to the solid slag in the distillation retort for 55 min, stirring and extracting for 2 times, filtering, combining the filtrates, cooling the filtrate through an ice bath, adjusting the pH to 4.0 by acidification of concentrated hydrochloric acid, standing for precipitation, filtering and collecting precipitates; extracting the mother liquor with ethyl acetate at volume ratio of 1: 0.6 for 2 times, recovering ethyl acetate, mixing the solid extracted with ethyl acetate and the precipitate to obtain crude extract, drying at low temperature, and weighing 6.52 kg. Mixing the crude extract with C18 bonded silica gel, and mixing the raw materials in parts by weight: the crude extract and C18 bonded silica gel are 1: 11, the wet method is used for loading the column, the dry method is used for loading, the eluent with the volume ratio of ethanol, glacial acetic acid and water being 7: 0.5: 2.5 is used for elution, HPLC detection is used for detecting, the same component at the position with 10.5 min of chromatogram retention time is intercepted, the eluent is distilled and recovered, ethyl acetate is used for recrystallization, and ethanol is used for secondary recrystallization, and other high-purity icariine and other flavonoid substances such as epimedin A, epimedin B, epimedin C and the like can be obtained, wherein the icariine content is 1.25 kg, the epimedin content is 98.9 percent, and the extraction rate exceeds 98.6 percent.
Example four:
drying herba Epimedii in shade, pulverizing with pulverizer, and sieving with 80 mesh sieve; weighing 90 kg of epimedium herb powder in a distillation retort, adding 1.45 times of sodium carbonate aqueous solution with the pH value of 10 in weight of the herb powder to ensure that the epimedium herb powder is completely and loosely floated, soaking for 40 min, introducing water vapor and stirring for 60 min, filtering while hot, introducing the water vapor into the solid slag in the distillation retort for 60 min, stirring and extracting for 2 times, filtering, combining filtrates, cooling the filtrate by an ice bath, acidifying with concentrated hydrochloric acid to adjust the pH value to 3.5, standing for precipitation, filtering and collecting precipitates; extracting the mother liquor with ethyl acetate at volume ratio of 1: 0.7 for 2 times, recovering ethyl acetate, mixing the solid extracted with ethyl acetate and the precipitate to obtain crude extract, drying at low temperature, and weighing 7.35 kg. Mixing the crude extract with C18 bonded silica gel, and mixing the raw materials in parts by weight: the crude extract and C18 bonded silica gel are 1: 12, wet-process column loading is carried out, dry-process sample loading is carried out, elution is carried out by eluent with the volume ratio of ethanol, glacial acetic acid and water being 7: 0.6: 2.4, HPLC detection is carried out, the same component at 10.5 min of chromatogram retention time is intercepted, eluent is distilled and recovered, ethyl acetate is used for recrystallization, and ethanol is used for secondary recrystallization, thus obtaining high-purity icariine and other flavonoid substances such as epimedin A, epimedin B, epimedin C and the like, wherein the icariine content is 1.41 kg, the epimedin content is 99.1%, and the extraction rate is 98.3%.
Claims (1)
1. A method for extracting icariin from epimedium herb is completed according to the following process steps,
(1) drying herba Epimedii in shade, pulverizing with pulverizer, and sieving with 80 mesh sieve;
(2) weighing 50 parts by weight of herba Epimedii whole plant powder, adding 1.4-1.5 times of sodium carbonate aqueous solution with pH of 10-11 in a distillation retort, soaking for 30-40 min, introducing water vapor, stirring for 50-60 min, and filtering while hot;
(3) introducing water vapor into the solid residue in a distillation tank for 50-60 min, stirring, and filtering;
(4) repeating the step (3) once;
(5) combining the filtrates from the step (2) to the step (4), cooling in an ice bath, adjusting the pH of the filtrate to 3.5-4.2 by using concentrated hydrochloric acid, standing for precipitation, filtering and collecting precipitates;
(6) extracting the mother liquor with ethyl acetate for 2 times, wherein the using amount of the ethyl acetate is 0.6-0.7 time of the volume of the mother liquor each time, recovering the ethyl acetate, combining the solid extracted with the ethyl acetate and the precipitate obtained in the step (5) to obtain a crude extract, and drying at low temperature;
(7) mixing the dried crude extract with C18 bonded silica gel, and mixing the mixture according to the weight ratio: the crude extract and C18 bonded silica gel are 1: 10-12, the wet method is used for column loading, the dry method is used for sample loading, the eluent with the ratio of ethanol to glacial acetic acid to water being 7: 0.5-0.6: 2.4-2.5 is used for elution, HPLC is used for detection, the retention time of the chromatogram of the icariin reference substance is 10.5 min, and the same components are intercepted;
(8) distilling to recover eluent, recrystallizing with ethyl acetate, and recrystallizing with ethanol.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2007031140A2 (en) * | 2005-07-15 | 2007-03-22 | Dr. Willmar Schwabe Gmbh & Co. Kg | Extracts from epimedium species method for production and use thereof |
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| CN101812100A (en) * | 2010-04-08 | 2010-08-25 | 苏州宝泽堂医药科技有限公司 | Method for preparing icariin |
| CN106589020A (en) * | 2016-12-31 | 2017-04-26 | 北京颐方生物科技有限公司 | Method for extracting icariin from epimedium |
| CN112266399A (en) * | 2020-09-28 | 2021-01-26 | 陕西天骄生物科技有限公司 | High-purity separation and extraction method of epimedium extract |
-
2021
- 2021-11-09 CN CN202111317176.6A patent/CN113980065A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007031140A2 (en) * | 2005-07-15 | 2007-03-22 | Dr. Willmar Schwabe Gmbh & Co. Kg | Extracts from epimedium species method for production and use thereof |
| CN101607964A (en) * | 2009-06-03 | 2009-12-23 | 贾晓斌 | A kind of preparation method of cycle epimedium aglucone |
| CN101812100A (en) * | 2010-04-08 | 2010-08-25 | 苏州宝泽堂医药科技有限公司 | Method for preparing icariin |
| CN106589020A (en) * | 2016-12-31 | 2017-04-26 | 北京颐方生物科技有限公司 | Method for extracting icariin from epimedium |
| CN112266399A (en) * | 2020-09-28 | 2021-01-26 | 陕西天骄生物科技有限公司 | High-purity separation and extraction method of epimedium extract |
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