CN102824394A - Method for synchronously extracting and separating icariin and icarisid II from herba epimedii - Google Patents
Method for synchronously extracting and separating icariin and icarisid II from herba epimedii Download PDFInfo
- Publication number
- CN102824394A CN102824394A CN2012103451028A CN201210345102A CN102824394A CN 102824394 A CN102824394 A CN 102824394A CN 2012103451028 A CN2012103451028 A CN 2012103451028A CN 201210345102 A CN201210345102 A CN 201210345102A CN 102824394 A CN102824394 A CN 102824394A
- Authority
- CN
- China
- Prior art keywords
- ethanol
- icariin
- eluent
- alcohol
- concentration
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a method for synchronously extracting and separating icariin and icarisid II from herba epimedii. The method obtains high-purity extract of icariin and icarisid II through macroporous adsorption resin gradient elution respectively. The invention also provides a method for extracting and separating icariin or icarisid II. The method provided by the invention not only realizes synchronous extraction and separation of icariin and icarisid II and simplifies the technological process, but also obtains a medicinal intermediate with relatively high purity, obviously increases the yield of monomer components, improves the utilization rate of medicinal materials and reduces the production cost; and meanwhile, without using organic solvents with relatively high toxicity, the method guarantees the safety of operators, realizes the green production and has great industrial application prospects.
Description
Technical field
The present invention relates to from Herba Epimedii the method for extraction separation icariin and icariside I I simultaneously.
Background technology
Herba Epimedii is a kind of traditional traditional tonic medicine, has multiple efficacies such as kidney-replenishing, bone and muscle strengthening, wind-damp dispelling.Because modern Natural Medicine Chemistry subject development is more and more to the research of the monomer reactivity composition in the Herba Epimedii.
Icariin is one of monomer of extraction separation from Herba Epimedii, for aspects such as improving cardiovascular system, endocrine regulation, enhancing immunity, gonadotropic Effect and antitumor, anti-liver poison positive curative effect effect is arranged all.Icariside I I is the new monomer of from Herba Epimedii, developing in recent years, improves at blood vessel inner skin cell function to be superior to icariin on; Aspect inhibition lipid peroxidation, TAC and reducing power, all has tangible oxidation resistance; Have the pharmacological action identical (strong bone, gonadal hormone appearance etc.) in addition equally with the icariin part.
Because the pharmacological action that icariin and icariside I I are outstanding, made it in treatment osteoporosis, immune system, cardiovascular and cerebrovascular vessel or anticancer, the medicine that gives protection against cancer or health product, occupy critical role, its Application and Development and research prospect are broader.
To the above-mentioned flavones ingredient of Herba Epimedii, have the report of related manufacturing processes at present.Like the patent No.: 03141371.4, in this patent the Herba Epimedii water extract-alcohol precipitation is received last D101 macroporous adsorbent resin; And carry out eluting with 85% ethanol; Finally obtain general flavone content at the Herba Epimedii extract more than 50%, wherein, icariin content can reach about 9%.But in the above-mentioned patent, in the extract, content Determination of Icariin is lower, if also need the further separation and purification to icariin, subsequent operation is more complicated, and cost is higher.Number of patent application: CN 200710133883.3, in this patent application, with Herba Epimedii with 70% ethanol extraction after; After dichloromethane takes off assorted, ethyl acetate extraction, go up the DM130 macroporous resin again, earlier with after aqueous slkali, the 20% alcohol flushing remove impurity; Again with 60% ethanol elution; After the drying, icariin content reaches 52% in the extract, and yield reaches 45.11%; This patent points out that also 55%-60% ethanol is best to the elute effect of icariin.This patent application method has effectively improved the purity of icariin, makes the later stage separation and purification more easy; But, in this method, need to adopt dichloromethane; It is a kind of toxic solvents, when high concentration, can cause damage to human liver, kidney and cerebral tissue, and; It has carcinogenesis to animal, is human potential carcinogen.For icariside I I; Mostly be at present to adopt the icariin hydrolysis is obtained; The report that does not have effective purification icariside I I from Herba Epimedii, the icariside I I though this mode can effectively be purified, this method is with the icariin hydrolysis; Can't obtain icariin simultaneously, be unfavorable for the utilization of herb resource.
Do not see at present and adopt same separation method direct separation to extract the relevant report of icariin and icariside I I.
Summary of the invention
The object of the present invention is to provide a kind of environmental safety good, from Herba Epimedii the method for extraction separation icariin and icariside I I simultaneously.Another object of the present invention is to provide the method for extraction separation icariin or icariside I I.
The invention provides a kind of from Herba Epimedii the method for extraction separation icariin and icariside I I simultaneously, it comprises following operating procedure:
(1) get epimedium herb, behind water extract-alcohol precipitation, with gained supernatant concentrating under reduced pressure, concentrated solution is subsequent use;
(2) get concentrated solution, go up nonpolar or Semi-polarity macroporous adsorptive resins, use the 20%-70%V/V ethanol elution successively, collecting concentration of alcohol respectively is that 40%-45%V/V eluent I and concentration of alcohol are the eluent II of 50%-70%V/V;
(3) with behind the eluent I recovery ethanol,, collect ethyl acetate layer, behind the recovery solvent, add the 50%-70%V/V dissolve with ethanol with water-ethyl acetate system extraction, filtration, filtrating cold preservation is left standstill, and separates out solid content, promptly gets the icariin bullion;
(4) with behind the eluent II recovery ethanol,, collect ethyl acetate layer, behind the recovery solvent, add the 50%-70%V/V dissolve with ethanol with water-ethyl acetate system extraction, filtration, filtrating cold preservation is left standstill, and separates out solid content, promptly gets icariside I I bullion.
Wherein, in the said water extract-alcohol precipitation process of step (1), concentration of alcohol is more than 60%V/V.
Further, in the said water extract-alcohol precipitation process of step (1), concentration of alcohol is 60%V/V.
Further, in the step (1), the concrete operations that said water is carried are following:
Get epimedium herb, decocte with water merges decocting liquid more than 2 times, and being evaporated to 60 ℃ of relative densities of measuring down is 1.1-1.2.
Wherein, in the step (2), said nonpolar macroporous adsorption resin is D101 type, AB-8 type or DM-130 type; Said Semi-polarity macroporous adsorbent resin is the DM301 type.
Further, the concrete operations of step (2) are following:
Get concentrated solution; Last DM301, DM-130 or AB-8 macroporous adsorptive resins; Use concentration to carry out eluting successively, and to collect eluent I and the concentration of alcohol that concentration of alcohol is 40%-45%V/V respectively be the eluent II of 50%-70%V/V as 20%V/V, 30%V/V, 40%V/V, 45%V/V, 50%V/V, 60%V/V, 70%V/V ethanol.
Further, in the step (2), the 20%V/V amount of ethanol is a 4-5 times of column volume; The 30%V/V amount of ethanol is a 3-5 times of column volume; The 40%V/V amount of ethanol is a 4-6 times of column volume; The 45%V/V amount of ethanol is a 3-5 times of column volume; The 50%V/V amount of ethanol is a 3-5 times of column volume; The 60%V/V amount of ethanol is a 4-5 times of column volume; The 70%V/V amount of ethanol is a 3-6 times of column volume.
Wherein, concentration of ethanol is 60%V/V described in step (3), (4), and the temperature of said cold preservation is 0 ~ 4 ℃.
Wherein, in the said icariin bullion, icariin content is 45%-50%W/W; In the said icariside I I bullion, icariside I I content is 58%-63%W/W.
Wherein, said Herba Epimedii is selected from Berberidaceae plant Herba Epimedii (Epimedium brevicornu Maxim.), Herba Epimedii (Epimedium koresnum Nakai.), arrow leaf Herba Epimedii (Epimedium sagittatum (Sieb.et Zucc.) Maxim.), pubescence Herba Epimedii (Epimedium pubescens Maxim.), (Epimedium wushanense T. (S.Ying) and nearly edge thereof are for using kind for Epimedium wushanense.
The present invention also provides a kind of method of extraction separation icariin, and it comprises following operating procedure:
(1) get epimedium herb, behind water extract-alcohol precipitation, with gained supernatant concentrating under reduced pressure, concentrated solution is subsequent use;
(2) get concentrated solution, go up nonpolar or Semi-polarity macroporous adsorptive resins, use the 20%-45%V/V ethanol elution successively, collecting concentration of alcohol respectively is the 40%-45%V/V eluent;
(3) eluent is reclaimed ethanol after, with the extraction of water-ethyl acetate system, collect ethyl acetate layer, reclaim solvent after, add the 60%V/V dissolve with ethanol, filter, filtrating cold preservation is left standstill, and separates out solid content, promptly gets the icariin bullion.
Further, the concrete operations of step (2) are following:
Get concentrated solution, last DM301, DM-130 or AB-8 macroporous adsorptive resins use concentration to carry out eluting as 20%V/V, 30%V/V, 40%V/V, 45%V/V ethanol successively, and collecting concentration of alcohol is the eluent of 40%-45%V/V.
The present invention also provides a kind of method of separating icariside I I, and it comprises following operating procedure:
(1) get epimedium herb, behind water extract-alcohol precipitation, with gained supernatant concentrating under reduced pressure, concentrated solution is subsequent use;
(2) get concentrated solution, go up nonpolar or Semi-polarity macroporous adsorptive resins, use 20%-45%V/V, 50%-70%V/V ethanol elution successively, collecting concentration of alcohol is the eluent of 50%-70%V/V;
(3) with eluent, behind the recovery ethanol,, collect ethyl acetate layer with water-ethyl acetate system extraction, behind the recovery solvent, add 60% dissolve with ethanol, to filter, the cold preservation of filtrating is left standstill, and separates out solid content, promptly gets icariside I I bullion.
Further, the concrete operations of step (2) are following:
Get concentrated solution, last DM301, DM-130 or AB-8 macroporous adsorptive resins use concentration to carry out eluting as 20%V/V, 30%V/V, 40%V/V, 45%V/V, 50%V/V, 60%V/V, 70%V/V ethanol successively, and collecting concentration of alcohol is the eluent of 50%-70%V/V.
Compared with prior art, the invention has the advantages that:
(1) the present invention can be under with a kind of separation method; Realize extraction separation simultaneously to icariin and two kinds of monomer components of icariside I I; Simplify the extraction separation flow process of two kinds of monomer components, made full use of the epimedium herb resource, significantly reduced production cost;
(2) in the extraction separation process of the present invention, do not use the bigger organic solvents of toxicity such as dichloromethane, avoided health hazard, reduced pollution environment to operator;
(3) the inventive method can obtain higher icariin bullion (purity 45%-50%) of purity and icariside I I bullion (purity 58%-63%) respectively, is that being further purified with medical applications of two kinds of monomer components provides high-load intermediate.Simultaneously, in the extraction separation method of the present invention, the yield of two kinds of monomer components obviously is superior to existing separation method all more than 80%.
Description of drawings
The liquid chromatogram of Fig. 1 icariin reference substance;
The liquid chromatogram of Fig. 2 icariside I I reference substance;
Fig. 3 icariin bullion liquid chromatogram;
The liquid chromatogram of Fig. 4 icariside I I bullion.
The specific embodiment
Used Herba Epimedii among the present invention; All with reference to " Chinese pharmacopoeia 2005 editions is selected from Berberidaceae plant Herba Epimedii (Epimedium brevicornu Maxim.), Herba Epimedii (Epimedium koresnum Nakai.), arrow leaf Herba Epimedii (Epimedium sagittatum (Sieb.et Zucc.) Maxim.), pubescence Herba Epimedii (Epimedium pubescens Maxim.), Epimedium wushanense (Epimedium wushanense T. (S.Ying).Its nearly edge is for also using with kind.
The separation method of embodiment 1 icariin of the present invention and icariside I I
1) gets Herba Epimedii (Epimedium koresnum Nakai.) medical material 500g simultaneously: measure icariin, moisture content.
2) with 1) described in medical material soak 15h with 17 times of water yields, decoct 2h, filter and obtain medicinal residues and filtrating respectively.
3) with 2) middle gained medicinal residues are with 10 times of water yields immersion 15h, and decoction 2h filters and obtains medicinal residues and filtrating for the second time respectively.
4) with 2) and 3) merging of middle gained filtrating, twice medicinal residues abandon.The filtrating that merges is left standstill 9h, get its supernatant at 55 ℃ of-70 ℃ of section vacuum rotary steams, be concentrated into 500ml, measuring relative density down at 60 ℃ is the 1.1-1.2 range section, obtains extractum.
5) with 4) in gained extractum to add 95% ethanol to mixed liquor concentration of alcohol be 60%, place 10h, centrifugal, get the supernatant distilling under reduced pressure to 500ml.
6) pretreatment obtains macroporous resin, and preprocess method is: with 450g DM301 resin, and 5% NaCl solution-treated supernatant, reuse distilled water cleaning many times, saturated 20h, subsequent use.
7) with 5) in the medicinal liquid of gained pack 6 into) in the macroporous resin column of gained, presaturation 22h obtains saturated macroporous resin again.
8) with 4 times of column volumes of 20% ethanol; 3 times of column volumes of 30% ethanol, 4 times of column volumes of 40% ethanol, 3 times of column volumes of 45% ethanol, 3 times of column volumes of 50% ethanol; 4 times of column volumes of 60% ethanol; 3 times of column volumes of 70% ethanol, 3 times of column volumes of 80% ethanol handle 7) the middle saturated macroporous resin of gained, obtain the variable concentrations eluent: 40% eluent, 45% eluent, 50% eluent, 60% eluent, 70% eluent.
9) with 8) in 40% eluent, 45% eluent, 50% eluent, 60% eluent, 70% eluent of gained carry out concentrating under reduced pressure respectively, reclaim ethanol and obtain the fluid extract of variable concentrations.
10) with 9) middle gained fluid extract heating, respectively get ethyl acetate 80ml, extract 10min respectively and discard water layer.Reclaim ethyl acetate.
11) will be through 10) extractum after handling adds each 30ml of 60% ethanol, and ultrasonic 21min filters, and filtering residue discards, filtrate for later use.
12) with 11) in gained filtrating spend the night 0 ~ 4 ℃ of cold preservation, deposition is separated out in crystallization.
13) with 12) in gained deposition merge; The merging of 40%-45% eluent obtains 46% icariin bullion (icariin 46%; Icariside I I 2.5%), the merging of 50%-70% eluent obtains 59% icariside I I bullion (icariin 0%, icariside I I 59%).
Gained finished product purity testing experimental technique is following:
Chromatographic condition: Dikma C
18Chromatographic column (4.5 * 150mm, 5 μ m); Mobile phase: acetonitrile (A)-water (B) gradient elution, 0 ~ 7min, 30%A, 7 ~ 8min, 30% ~ 45%A, 8 ~ 20min, 45%A; Flow velocity: 1.0ml/min; Column temperature: 30 ℃; Detect wavelength: 270nm.
The preparation of reference substance solution: get icariin respectively, icariside I I reference substance is an amount of, accurately claim surely, add methanol and process the solution that every 1ml contains 0.1mg approximately, promptly get.
The preparation of need testing solution: get the about 0.1g of finished product, the accurate title, decide, and puts in the tool plug measuring bottle, adds Diluted Alcohol 50ml dissolving, shakes up, and filters, and gets subsequent filtrate, promptly gets.
Algoscopy: get each 10 μ l of above-mentioned two kinds of solution respectively, inject chromatograph of liquid, measure, promptly get.
Measure the result and see Fig. 1-4.
The separation method of embodiment 2 icariin of the present invention and icariside I I
1) gets Herba Epimedii (Epimedium brevicornu Maxim.) medical material 500g simultaneously: measure icariin, moisture content.
2) with 1) described in medical material soak 15h with 19 times of water yields, decoct 2h, filter and obtain medicinal residues and filtrating respectively.
3) with 2) middle gained medicinal residues are with 11 times of water yields immersion 15h, and decoction 2h filters and obtains medicinal residues and filtrating for the second time respectively.
4) with 2) and 3) merging of middle gained filtrating, twice medicinal residues abandon.The filtrating that merges is left standstill 13h, get its supernatant at 55 ℃ of-70 ℃ of section vacuum rotary steams, be concentrated into 500ml, measuring relative density down at 60 ℃ is the 1.1-1.2 range section, obtains extractum.
5) with 4) in gained extractum to add 95% ethanol to mixed liquor concentration of alcohol be 60%, place 8h, centrifugal, get the supernatant distilling under reduced pressure to 500ml.
6) pretreatment obtains macroporous resin, and preprocess method is: with 450g AB-8 resin, and 5% NaCl solution-treated supernatant, reuse distilled water cleaning many times, saturated 18h, subsequent use.
7) with 5) in the medicinal liquid of gained pack 6 into) in the macroporous resin column of gained, presaturation 24h obtains saturated macroporous resin again.
8) with 5 times of column volumes of 20% ethanol, 5 times of column volumes of 30% ethanol, 6 times of column volumes of 40% ethanol; 4 times of column volumes of 45% ethanol; 5 times of column volumes of 50% ethanol, 4 times of column volumes of 60% ethanol, 6 times of column volumes of 70% ethanol; 4 times of column volumes of 80% ethanol handle 7) the middle saturated macroporous resin of gained, obtain the variable concentrations eluent: 40% eluent, 45% eluent, 50% eluent, 60% eluent, 70% eluent.
9) with 8) in 40% eluent, 45% eluent, 50% eluent, 60% eluent, 70% eluent of gained carry out concentrating under reduced pressure respectively, reclaim ethanol and obtain the fluid extract of variable concentrations.
10) with 9) middle gained fluid extract heating, respectively get ethyl acetate 100ml, extract 12min respectively and discard water layer.Reclaim ethyl acetate.
11) will be through 10) extractum after handling adds each 50ml of 60% ethanol, and ultrasonic 18min filters, and filtering residue discards, filtrate for later use.
12) with 11) in gained filtrating spend the night 0 ~ 4 ℃ of cold preservation, deposition is separated out in crystallization.
13) with 12) in gained deposition merge, the 40%-45% eluent merges and obtains 49% icariin bullion, the 50%-70% eluent merges and obtains 61% icariside I I bullion.Assay method such as embodiment 1.
The separation method of embodiment 3 icariin of the present invention and icariside I I
1) gets arrow leaf Herba Epimedii (Epimedium sagittatum (Sieb.et Zucc.) Maxim.) medical material 500g simultaneously: measure icariin, moisture content.
2) with 1) described in medical material soak 15h with 19 times of water yields, decoct 2h, filter and obtain medicinal residues and filtrating respectively.
3) with 2) middle gained medicinal residues are with 7 times of water yields immersion 15h, and decoction 2h filters and obtains medicinal residues and filtrating for the second time respectively.
4) with 2) and 3) merging of middle gained filtrating, twice medicinal residues abandon.The filtrating that merges is left standstill 11h, get its supernatant at 55 ℃ of-70 ℃ of section vacuum rotary steams, be concentrated into 500ml, measuring relative density down at 60 ℃ is the 1.1-1.2 range section, obtains extractum.
5) with 4) in gained extractum to add 95% ethanol to mixed liquor concentration of alcohol be 60%, place 6h, centrifugal, get the supernatant distilling under reduced pressure to 500ml.
6) pretreatment obtains macroporous resin, and preprocess method is: with 450g DM301 resin, and 5% NaCl solution-treated supernatant, reuse distilled water cleaning many times, saturated 22h, subsequent use.
7) with 5) in the medicinal liquid of gained pack 6 into) in the macroporous resin column of gained, presaturation 26h obtains saturated macroporous resin again.
8) with 6 times of column volumes of 20% ethanol, 7 times of column volumes of 30% ethanol, 5 times of column volumes of 40% ethanol; 5 times of column volumes of 45% ethanol; 7 times of column volumes of 50% ethanol, 6 times of column volumes of 60% ethanol, 6 times of column volumes of 70% ethanol; 4 times of column volumes of 80% ethanol handle 7) the middle saturated macroporous resin of gained, obtain the variable concentrations eluent: 40% eluent, 45% eluent, 50% eluent, 60% eluent, 70% eluent.
9) with 8) in 40% eluent, 45% eluent, 50% eluent, 60% eluent, 70% eluent of gained carry out concentrating under reduced pressure respectively, reclaim ethanol and obtain the fluid extract of variable concentrations.
10) with 9) middle gained fluid extract heating, respectively get ethyl acetate 120ml, extract 8min respectively and discard water layer.Reclaim ethyl acetate.
11) will be through 10) extractum after handling adds each 70ml of 60% ethanol, and ultrasonic 22min filters, and filtering residue discards, filtrate for later use.
12) with 11) in gained filtrating spend the night 0 ~ 4 ℃ of cold preservation, deposition is separated out in crystallization.
13) with 12) in gained deposition merge, the 40%-45% eluent merges and obtains 48% icariin bullion, the 50%-70% eluent merges and obtains 62% icariside I I bullion.Assay method such as embodiment 1.
Through measuring, in the extraction separation method of embodiment of the invention 1-3, the yield of icariin is about 80%, and the yield of icariside I I is about 85%.
The screening of embodiment 4 macroporous resin models
Macroporous resin preprocess method:, subsequent use according to handling with method among the embodiment 1.
The test of static adsorption/desorption: get each 1g of macroporous resin that handles well, place 50ml tool plug conical flask respectively, add the embodiment 1 step 5) gained solution of 20ml, leave standstill 24h, inclining the residual liquid after the absorption.Macroporous resin with water washing after, attach with the Different concentrations of alcohol solution stripping.Press assay method and measure 5) in the concentration of solution, residual liquid and stripping liquid, and be index with icariin and icariside I I total amount, press examination and calculate adsorption capacity and desorption efficiency, the result sees table 1.
Table 1
Can be known that by table 1 DM301, DM-130 and AB-8 are all better to the adsorptive value and the desorption efficiency of icariin and icariside I I total amount, resin is used in the separation that all can be used as icariin and icariside I I; And the D101 resin also can be used for the separation of mentioned component, but its adsorptive value is lower, is unfavorable for the saving of resource, and therefore, the present invention is with preferred DM301, DM-130 and AB-8 type macroporous adsorbent resin.
In sum, extraction separation when the present invention has not only realized icariin and icariside I I has been simplified technological process; Also obtained the higher pharmaceutical intermediate of purity; Significantly increase the yield of monomer component, improved the utilization rate of medical material, reduced production cost; Simultaneously, the inventive method is not used the bigger organic solvent of toxicity, has ensured safety of operators, has realized green production, has favorable industrial application prospect.
Claims (12)
1. the method for extraction separation icariin and icariside I I simultaneously from Herba Epimedii, it is characterized in that: it comprises following operating procedure:
(1) get epimedium herb, behind water extract-alcohol precipitation, with gained supernatant concentrating under reduced pressure, concentrated solution is subsequent use;
(2) get concentrated solution, go up nonpolar or Semi-polarity macroporous adsorptive resins, use the 20%-70%V/V ethanol elution successively, collecting concentration of alcohol respectively is that 40%-45%V/V eluent I and concentration of alcohol are the eluent II of 50%-70%V/V;
(3) with behind the eluent I recovery ethanol,, collect ethyl acetate layer, behind the recovery solvent, add the 50%-70%V/V dissolve with ethanol with water-ethyl acetate system extraction, filtration, filtrating cold preservation is left standstill, and separates out solid content, promptly gets the icariin bullion;
(4) with the eluent II, behind the recovery ethanol,, collect ethyl acetate layer with water-ethyl acetate system extraction, behind the recovery solvent, add the 50%-70%V/V dissolve with ethanol, filtration, filtrating cold preservation is left standstill, and separates out solid content, promptly gets icariside I I bullion.
2. method according to claim 1 is characterized in that: in the said water extract-alcohol precipitation process of step (1), concentration of alcohol is more than 60%V/V.
3. method according to claim 2 is characterized in that: in the said water extract-alcohol precipitation process of step (1), concentration of alcohol is 60%V/V.
4. according to any described method of claim 1-3, it is characterized in that: in the step (1), the concrete operations that said water is carried are following:
Get epimedium herb, decocte with water merges decocting liquid more than 2 times, and being evaporated to 60 ℃ of relative densities of measuring down is 1.1-1.2.
5. method according to claim 1 is characterized in that: in the step (2), said nonpolar macroporous adsorption resin is D101 type, AB-8 type or DM-130 type; Said Semi-polarity macroporous adsorbent resin is the DM301 type.
6. method according to claim 5 is characterized in that: the concrete operations of step (2) are following:
Get concentrated solution; Last DM301, DM-130 or AB-8 macroporous adsorptive resins; Use concentration to carry out eluting successively, and to collect eluent I and the concentration of alcohol that concentration of alcohol is 40%-45%V/V respectively be the eluent II of 50%-70%V/V as 20%V/V, 30%V/V, 40%V/V, 45%V/V, 50%V/V, 60%V/V, 70%V/V ethanol.
7. method according to claim 6 is characterized in that: in the step (2), the 20%V/V amount of ethanol is a 4-5 times of column volume; The 30%V/V amount of ethanol is a 3-5 times of column volume; The 40%V/V amount of ethanol is a 4-6 times of column volume; The 45%V/V amount of ethanol is a 3-5 times of column volume; The 50%V/V amount of ethanol is a 3-5 times of column volume; The 60%V/V amount of ethanol is a 4-5 times of column volume; The 70%V/V amount of ethanol is a 3-6 times of column volume.
8. method according to claim 1 is characterized in that: concentration of alcohol is 60%V/V described in step (3), (4), and the temperature of said cold preservation is 0 ~ 4 ℃.
9. according to any described method of claim 1-8, it is characterized in that: in the said icariin bullion, icariin content is 45%-50%W/W; In the said icariside I I bullion, icariside I I content is 58%-63%W/W.
10. according to any described method of claim 1-9, it is characterized in that: said Herba Epimedii is selected from Berberidaceae plant Herba Epimedii (Epimedium brevicornu Maxim.), Herba Epimedii (Epimedium koresnum Nakai.), arrow leaf Herba Epimedii (Epimedium sagittatum (Sieb.et Zucc.) Maxim.), pubescence Herba Epimedii (Epimedium pubescens Maxim.), (Epimedium wushanense T. (S.Ying) and nearly edge thereof are for using kind for Epimedium wushanense.
11. the method for an extraction separation icariin is characterized in that: it comprises following operating procedure:
(1) get epimedium herb, behind water extract-alcohol precipitation, with gained supernatant concentrating under reduced pressure, concentrated solution is subsequent use;
(2) get concentrated solution, go up nonpolar or Semi-polarity macroporous adsorptive resins, use the 20%-45%V/V ethanol elution successively, collecting concentration of alcohol is the 40%-45%V/V eluent;
(3) eluent is reclaimed ethanol after, with the extraction of water-ethyl acetate system, collect ethyl acetate layer, reclaim solvent after, add the 60%V/V dissolve with ethanol, filter, filtrating cold preservation is left standstill, and separates out solid content, promptly gets the icariin bullion.
12. the method for an extraction separation icariside I I is characterized in that: it comprises following operating procedure:
(1) get epimedium herb, behind water extract-alcohol precipitation, with gained supernatant concentrating under reduced pressure, concentrated solution is subsequent use;
(2) get concentrated solution, go up nonpolar or Semi-polarity macroporous adsorptive resins, use 20%-45%V/V, 50%-70%V/V ethanol elution successively, collecting concentration of alcohol is the eluent of 50%-70%V/V;
(3) with eluent, behind the recovery ethanol,, collect ethyl acetate layer with water-ethyl acetate system extraction, behind the recovery solvent, add the 60%V/V dissolve with ethanol, filtration, filtrating cold preservation is left standstill, and separates out solid content, promptly gets icariside I I bullion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210345102.8A CN102824394B (en) | 2012-09-18 | 2012-09-18 | Method for synchronously extracting and separating icariin and icarisid II from herba epimedii |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210345102.8A CN102824394B (en) | 2012-09-18 | 2012-09-18 | Method for synchronously extracting and separating icariin and icarisid II from herba epimedii |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102824394A true CN102824394A (en) | 2012-12-19 |
| CN102824394B CN102824394B (en) | 2014-06-04 |
Family
ID=47327840
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210345102.8A Expired - Fee Related CN102824394B (en) | 2012-09-18 | 2012-09-18 | Method for synchronously extracting and separating icariin and icarisid II from herba epimedii |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102824394B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104910216A (en) * | 2015-03-07 | 2015-09-16 | 宝鸡文理学院 | Separation method for obtaining a plurality of epimeddium flavones by preparative liquid chromatography |
| CN105079013A (en) * | 2014-04-23 | 2015-11-25 | 北京东方百奥医药开发有限公司 | Uses of icariside II in preparation of products for prevention and treatment of reproduction dysfunction |
| CN106148449A (en) * | 2015-03-24 | 2016-11-23 | 北京珅奥基医药科技有限公司 | A kind of preparation method of icariside I |
| CN106831913A (en) * | 2017-02-15 | 2017-06-13 | 鲁南制药集团股份有限公司 | A kind of preparation technology of icariin |
| 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 |
| CN114790222A (en) * | 2022-05-11 | 2022-07-26 | 遵义医科大学 | Flavonoid compound based on epimedium herb and preparation method thereof |
-
2012
- 2012-09-18 CN CN201210345102.8A patent/CN102824394B/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 张鑫: "炮制对淫羊藿黄酮类成分的影响", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 雷永涛等: "淫羊藿总黄酮的提取和分离纯化工艺研究", 《贵阳医学院学报》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105079013A (en) * | 2014-04-23 | 2015-11-25 | 北京东方百奥医药开发有限公司 | Uses of icariside II in preparation of products for prevention and treatment of reproduction dysfunction |
| CN105079013B (en) * | 2014-04-23 | 2019-08-30 | 苏州广奥医药开发有限公司 | Purposes of the icariside II in the product of preparation prevention and treatment reproductive dysfunction |
| CN104910216A (en) * | 2015-03-07 | 2015-09-16 | 宝鸡文理学院 | Separation method for obtaining a plurality of epimeddium flavones by preparative liquid chromatography |
| CN104910216B (en) * | 2015-03-07 | 2017-11-03 | 宝鸡文理学院 | It is a kind of with preparing liquid phase method while obtaining the separation method of a variety of epimedium flavones |
| CN106148449A (en) * | 2015-03-24 | 2016-11-23 | 北京珅奥基医药科技有限公司 | A kind of preparation method of icariside I |
| CN106148449B (en) * | 2015-03-24 | 2020-12-25 | 北京珅奥基医药科技有限公司 | Preparation method of icariside I |
| CN106831913A (en) * | 2017-02-15 | 2017-06-13 | 鲁南制药集团股份有限公司 | A kind of preparation technology of icariin |
| 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 |
| CN112266399B (en) * | 2020-09-28 | 2023-11-10 | 陕西天骄生物科技有限公司 | High-purity separation and extraction method of epimedium extract |
| CN114790222A (en) * | 2022-05-11 | 2022-07-26 | 遵义医科大学 | Flavonoid compound based on epimedium herb and preparation method thereof |
| CN114790222B (en) * | 2022-05-11 | 2024-03-01 | 遵义医科大学 | Flavonoids based on epimedium and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102824394B (en) | 2014-06-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102824394A (en) | Method for synchronously extracting and separating icariin and icarisid II from herba epimedii | |
| CN104610410B (en) | A kind of extraction process of ginsenoside Rd | |
| CN106588616B (en) | A kind of preparation method of curcumin | |
| CN104592341A (en) | Method for extracting asiaticoside and madecassoside from centella | |
| CN102924537B (en) | Method for preparing hyperoside and isoquercitrin simultaneously from dogbane leaves | |
| CN104910216B (en) | It is a kind of with preparing liquid phase method while obtaining the separation method of a variety of epimedium flavones | |
| CN104817445B (en) | A kind of isolated and purified physcione and method of rheum emodin from Rhizoma Polygoni Cuspidati | |
| CN104370895B (en) | A kind of preparation method of orientin and Lutonaretin | |
| CN102659903A (en) | Method for extracting and purifying Chinese thorowax root saponin a or Chinese thorowax root saponin d | |
| CN101658598B (en) | Method for extracting and enriching alisma total triterpenic ketone alcohol components from alisma | |
| CN104262231B (en) | From white thorn seed, extract the method that separates L-Trp | |
| CN102180934A (en) | Preparation method of tubeimoside I | |
| CN102250183B (en) | Method for preparing high-purity ginsenoside Re by using ginseng flower buds as raw materials | |
| CN104473981B (en) | A kind of isolation and purification method of high-purity autumn eggplant leaf flavonoids | |
| CN106946833A (en) | A kind of method that high-purity sinensetin is extracted from Mao Xu Cao | |
| CN105906687B (en) | A kind of method that a variety of tanshinone monomer components are isolated and purified from the red sage root | |
| CN103288898B (en) | Neomangiferin reference substance extracts method for purifying and separating | |
| CN105061212B (en) | A kind of preparation method of neochlorogenic acid | |
| CN103739648A (en) | Preparation method for mussaendoside U | |
| CN104725449A (en) | Paeoniflorin extracting method | |
| CN102911218B (en) | Method for synchronously separating liquiritin and liquiritin apioside from liquorice | |
| CN103232468A (en) | Method for extracting purified oridonin from rabdosia rubescens | |
| CN103319546B (en) | A kind of from glutinous rehmannia the method for Separation of Water threose | |
| CN102050843B (en) | Method for extracting alkaloid and catechin from tea | |
| CN103739649A (en) | Preparation method for mussaendoside G |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140604 Termination date: 20140918 |
|
| EXPY | Termination of patent right or utility model |