CN1225470C - Method for extracting icariine from epimedium plant - Google Patents
Method for extracting icariine from epimedium plant Download PDFInfo
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Abstract
The present invention relates to a method for extracting icariin from barrenwort. The method has the technical key that by using chromatographic technology under normal pressure conditions, icariin crude products leached into the barrenwort are separated and purified; finally, an icariin pure product is obtained by recrystallization. The method provided by the present invention can separate and purify the icariin with high efficiency, and accordingly the icariin pure product with high purity and high recovery rate is obtained. A low-toxic organic solvent is used to avoid the residue of toxic and harmful organic solvents. Production technology is simple and feasible and has short time consumption. Used resources can be reused, and production cost is greatly reduced.
Description
Technical field
The present invention relates to a kind of method of from the Herba Epimedii plant, extracting icariine.
Background technology
Herba Epimedii is the over-ground part of Berberidaceae (Berberidaceae) Epimedium (Epimedium) plant, because barrenwort has kind more than 20 in China, account for the over half of global resources, and many compositions wherein can be medicinal, so extensive studies has been carried out for the pharmaceutical use that fully utilizes barrenwort in the home and abroad in recent years.So far, isolated more than 20 kind of monomeric compound from this platymiscium, it is broadly divided into four types from structure, that is: flavonoid glycosides, lignanoids, alkaloids and other class.Icariine is that its molecular formula is C by a kind of 8-isoamylene radical chromocor alcohol glycosides compound of separating in the crude drug
33H
40O
15, yellow needle crystal, molecular weight 676.67, fusing point 225-230 ℃.Pharmacological research shows that the physiologically active of icariine mainly shows and improves cardio-cerebrovascular function, enhancing body immunizing power, the synthetic and aspects such as prevention and therapeutic dysfunction of promotion DNA.Secondly, icariine also has anti-ageing and antitumor action.
Usually, all be to adopt from barrenwort, to extract icariine.Xu Suixu discloses from Herba Epimedii the method for extracting icariine in its patent CN1283627, comprise adopting soaking, decoct the back residue obtained liquid, extracting solution is concentrated after macroporous resin adsorption, and ethanol elution is with steps such as activated carbon decolorizings.This method causes the rate of recovery low because multistep is operated, the cost height, and can not obtain highly purified icariine.Peng Shulin discloses a kind of extraction process of Herba Epimedii total flavones in its patent CN1294132, utilize absorption one wash-out principle, earlier icariine is adsorbed onto on the macroporous resin from its water extract, again with ethanol with its wash-out, obtain content greater than 50% icariine crude product.Because what this method adopted is traditional absorbing process, resolving power is difficult to guarantee, so the product purity that obtains is low, is not easy to further use.In sum, the main problem that exists is in existing extracting method: 1, traditional absorption-desorption technology of being to use of its key problem in technology, and product purity is low; 2, purification step is many, loses greatly in the middle of the purpose component, and the rate of recovery is low; 3, some method causes it residual in product owing to used poisonous and hazardous organic solvent such as chloroform.
Summary of the invention
The objective of the invention is to overcome the product purity that exists in the existing processing method and the rate of recovery is low, the defective of residual harmful substance in the product, thereby provide a kind of chromatographic technique that utilizes, use the organic solvent of low toxicity, the method of high efficiency separation purification icariine is to obtain the pure product of icariine of high purity and high-recovery.
The objective of the invention is to realize by following technical scheme:
The invention provides a kind of method of extracting icariine from barrenwort, its processing step is as follows:
1) lixiviate icariine from barrenwort: after the raw material leaf pulverized, with the aqueous solution (I) refluxing extraction of the organic solvent of 15～20 times of amounts of its weight; Filter then, with the aqueous solution (I) the washing filter residue several of organic solvent; Merging filtrate and washings, concentrating under reduced pressure obtains the icariine crude product; The content of organic solvent is 50～100v% in the aqueous solution of described organic solvent (I); Described organic solvent comprises methyl alcohol, ethanol, Virahol, propyl carbinol, acetone, methyl acetate, ethyl acetate;
2) just separate icariine: the polarity or the low-pole macro-reticular adsorbing resin of non-ionic type are placed the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; Resin after this activation is adorned post with settling methods; As moving phase, the balance chromatography column is gone up sample with the icariine crude product that step 1) obtains with the aqueous solution (II) the dissolving back of organic solvent with the aqueous solution (II) of organic solvent; The aqueous solution (III) wash-out with organic solvent; Use the ultraviolet monitoring elution process, detect, collect second elution peak at 270nm; The elutriant that concentrating under reduced pressure is collected obtains icariine separated product just; The content of organic solvent is 5～25v% in the aqueous solution of described organic solvent (II); The content of organic solvent is 45～70v% in the aqueous solution of described organic solvent (III); Described organic solvent comprises methyl alcohol, ethanol, Virahol and acetone;
3) the first separated product of purification icariine: will be the inverted medium of matrix with silica gel or be that the inverted medium of matrix is adorned post with settling methods with the polymkeric substance; As moving phase, the balance chromatography column is with step with the aqueous solution (IV) of organic solvent
2) icariine that obtains just separated product is gone up sample with the aqueous solution (IV) the dissolving back of organic solvent; With the aqueous solution (V) of organic solvent as elutriant, gradient elution; Use the ultraviolet monitoring elution process, detect, collect the 3rd elution peak at 270nm; The elutriant that concentrating under reduced pressure is collected obtains icariine separated product again; The content of organic solvent is 35～55v% in the aqueous solution of described organic solvent (IV); The content of organic solvent is 40～80v% in the aqueous solution of described organic solvent (V); Described organic solvent comprises methyl alcohol, ethanol and Virahol;
4) recrystallization: the icariine that step 3) is obtained with the methyl alcohol of 35～45v% or ethanol separated product again carries out recrystallization, obtains the pure product of icariine.
The barrenwort of described step 1) comprises Da Ye Herba Epimedii, lobus cardiacus Herba Epimedii, arrow leaf Herba Epimedii, pubescence Herba Epimedii, Herba Epimedii, Da Hua Herba Epimedii and Epimedium wushanense.
The number of times of the refluxing extraction in the described step 1) is 1～4 time; The time of described refluxing extraction is 0.5～2 hour.
Described step 2) resin in comprises polystyrene, polyacrylamide, polymethacrylate, acrylamide and cinnamic multipolymer, or methacrylic ester and cinnamic multipolymer.
In the described step 3) is that the inverted medium of matrix is commercially available C with silica gel
8And C
18Inverted medium; In the described step 3) is that the inverted medium of matrix comprises that with polystyrene, polyacrylic ester, styrene diethylene benzene copoly mer, alkyl-modified polystyrene alcohol be the inverted medium of matrix with the polymkeric substance.
The method of extracting icariine from barrenwort provided by the invention is different from traditional absorbing process, and it utilizes chromatographic technique, thereby has following advantage:
A) product can reach the very high purity and the rate of recovery;
B) avoided poisonous and hazardous organic solvent residual in the product;
C) production technique is simple and direct feasible, and is consuming time shorter, and used resource reproducible utilization, greatly reduces production cost.
Description of drawings
Fig. 1 is the HPLC collection of illustrative plates of the icariine crude product that uses extracting method provided by the invention and obtain in step 1);
Fig. 2 is the icariine that uses extracting method provided by the invention and obtain in the step 3) HPLC collection of illustrative plates of separated product again;
Wherein peak 2 is corresponding to icariine.
Embodiment
1) after the Da Ye Herba Epimedii grassland material leaf that 20g is contained icariine 0.56% is pulverized, puts into the 1000ml matrass, add 300ml 50v% aqueous ethanolic solution, refluxed 1 hour; Incline supernatant liquid after, add 300ml 50v% aqueous ethanolic solution again to resistates, refluxed again 1 hour; Filter, for several times with same aqueous ethanolic solution washing filter residue; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 4.03g, and the icariine content of measuring wherein through HPLC is 2.78wt%, and the rate of recovery is 100%;
2) NKA-9 resin (Chemical Plant of Nankai Univ. produces, and component is a polystyrene) is placed the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; With this chromatography column of 25v% aqueous ethanolic solution balance; The icariine crude product 4.0g that step 1) is obtained fully dissolves upward sample of back with 15ml 25v% ethanol, after the abundant drip washing of 25v% aqueous ethanolic solution, with 65v% aqueous ethanolic solution wash-out; UV270nm detects, and collects second elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 363.3mg just, measures wherein through HPLC that icariine content is 30wt%, and the rate of recovery is 98%;
3) with PolyspherPST 10 (MERCK company produce, component is a styrene diethylene benzene copoly mer) inverted medium with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 45v% aqueous ethanolic solution balance; With step 2) icariine that obtains just separated product 360mg fully dissolve the last sample in back with 8ml 45% aqueous ethanolic solution, with 50～80v% aqueous ethanolic solution gradient elution; UV270nm detects, and collects the 3rd elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 122.7mg again, measures wherein through HPLC that icariine content is 88wt%, and the rate of recovery is 100%;
4) the 122.7mg icariine that step 3) is obtained with the methanol aqueous solution of 40v% again separated product carry out recrystallization, obtain the pure product 100.2mg of icariine, purity is 97%, the rate of recovery is 90%.
Embodiment 2, be raw material, separate the pure product of icariine that the preparation of purifying contains icariine 97.5% with the over-ground part of the lobus cardiacus Herba Epimedii grass that contains icariine 0.44%
1) after the lobus cardiacus Herba Epimedii grassland material leaf that 10g is contained icariine 0.44% is pulverized, puts into the 500ml matrass,, extract, use ethyl acetate 200ml at every turn, refluxed common refluxing extraction 3 times 1.5 hours with ethyl acetate backflow as same quadrat method among the embodiment 1; Filter, for several times with ethyl acetate washing filter residue; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 1.7g, and the icariine content of measuring wherein through HPLC is 2.63wt%, and the rate of recovery is 100%;
2) X-5 resin (Chemical Plant of Nankai Univ. produces, and component is acrylamide and cinnamic multipolymer) is placed the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; With this chromatography column of 20v% aqueous ethanolic solution balance; The icariine crude product 1.5g that step 1) is obtained fully dissolves upward sample of back with 10ml 20v% ethanol, after the abundant drip washing of 20v% aqueous ethanolic solution, with 60v% aqueous ethanolic solution wash-out; UV270nm detects, and collects second elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 121.4mg just, measures wherein through HPLC that icariine content is 32wt%, and the rate of recovery is 98.5%;
3) with C
18Inverted medium is with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 35v% methanol aqueous solution balance; With step 2) icariine that obtains just separated product 120mg fully dissolve the back with 5ml 35% methanol aqueous solution and go up sample, after the abundant drip washing of 35v% methanol aqueous solution, with 40～80v% methanol aqueous solution gradient elution; UV270nm detects, and collects the 3rd elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 39.0mg again, measures wherein through HPLC that icariine content is 88.5wt%, and the rate of recovery is 98%;
4) the 42.5mg icariine that step 3) is obtained with the methanol aqueous solution of 45v% again separated product carry out recrystallization, obtain the pure product 35.5mg of icariine, purity is 97.5%, the rate of recovery is 92%.
Embodiment 3, be raw material, separate the pure product of icariine that the preparation of purifying contains icariine 98% with the over-ground part of the arrow leaf Herba Epimedii grass that contains icariine 0.52%
1) after the arrow leaf Herba Epimedii grassland material leaf that 30g is contained icariine 0.52% is pulverized, puts into the 1000ml matrass, add the methanol aqueous solution of 350ml 60v%, refluxed 2 hours, filter, wash the filter residue several with same methanol aqueous solution; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 5.74g, and the icariine content of measuring wherein through HPLC is 2.72wt%, and the rate of recovery is 100%;
2) with XAD-4 (Rohm ﹠amp; Hass company produces, and component is methacrylic ester and cinnamic multipolymer) place the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; With this chromatography column of 25v% isopropanol water solution balance; The icariine crude product 5.0g that step 1) is obtained fully dissolves upward sample of back with 20ml 25v% isopropanol water solution, after the abundant drip washing of 25v% isopropanol water solution, with 45v% isopropanol water solution wash-out; UV270nm detects, and collects second elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 444.3mg just, measures wherein through HPLC that icariine content is 31wt%, and the rate of recovery is 98.5%;
3) with Oligo R3 (PE Biosystems company produce, component is alkyl-modified polystyrene alcohol) inverted medium with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 40v% methanol aqueous solution balance; With step 2) icariine that obtains just separated product 400mg fully dissolve the back with 15ml 40% methanol aqueous solution and go up sample, after the abundant drip washing of 40v% methanol aqueous solution, with 45～70v% methanol aqueous solution gradient elution; UV270nm detects, and collects the 3rd elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 134.7mg again, measures wherein through HPLC that icariine content is 92wt%, and the rate of recovery is 100%;
4) the 134.7mg icariine that step 3) is obtained with the aqueous ethanolic solution of 40v% again separated product carry out recrystallization, obtain the pure product 113.2mg of icariine, purity is 98.5%, the rate of recovery is 90%.
Embodiment 4, be raw material, separate the pure product of icariine that the preparation of purifying contains icariine 97% with the over-ground part of the pubescence Herba Epimedii grass that contains icariine 0.44%
1) after the pubescence Herba Epimedii grassland material leaf that 20g is contained icariine 0.44% is pulverized, puts into the 500ml matrass,, use the methyl acetate refluxing extraction, use methyl acetate 200ml at every turn, refluxed common refluxing extraction 4 times 0.5 hour as same quadrat method among the embodiment 1; Filter, for several times with methyl acetate washing filter residue; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 3.52g, and the icariine content of measuring wherein through HPLC is 2.50wt%, and the rate of recovery is 100%;
2) with XAD-10 (Rohm ﹠amp; Hass company produces, and component is a polyacrylamide) place the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; With this chromatography column of 5v% methanol aqueous solution balance; The icariine crude product 2.4g that step 1) is obtained fully dissolves upward sample of back with 25ml 5v% methyl alcohol, after the abundant drip washing of 5v% methanol aqueous solution, with 35v% methanol aqueous solution wash-out; UV270nm detects, and collects second elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 181.8mg just, measures wherein through HPLC that icariine content is 32wt%, and the rate of recovery is 97%;
3) with C
8Inverted medium is with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 35v% methanol aqueous solution balance; With step 2) icariine that obtains just separated product 180.0mg fully dissolve the last sample in back with 8ml 35% methanol aqueous solution, with 40～70v% methanol aqueous solution gradient elution; UV270nm detects, and collects the 3rd elution peak; The concentrating under reduced pressure elutriant obtains icariine separated product 61.9mg again, measures wherein through HPLC that icariine content is 89wt%, and the rate of recovery is 95.6%;
4) the 61.9mg icariine that step 3) is obtained with the aqueous ethanolic solution of 35v% again separated product carry out recrystallization, obtain the pure product 52.5mg of icariine, purity is 97.5%, the rate of recovery is 92.9%.
1) after the Herba Epimedii grassland material leaf that 30g is contained icariine 0.48% is pulverized, puts into the 1000ml matrass, add 300ml 20v% n-butanol aqueous solution, refluxed 2 hours, filter, wash the filter residue several with same n-butanol aqueous solution; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 5.41g, and the icariine content of measuring wherein through HPLC is 2.62wt%, and the rate of recovery is 98.5%;
2) with XAD-7 (Rohm ﹠amp; Hass company produces, and component is a polymethacrylate) place the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; With this chromatography column of 25v% aqueous acetone solution balance; The icariine crude product 5.4g that step 1) is obtained fully dissolves upward sample of back with 20ml 25v% aqueous acetone solution, after the abundant drip washing of 25v% aqueous acetone solution, and 50v% aqueous acetone solution wash-out; UV270nm detects, and collects second elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 443.2mg just, measures wherein through HPLC that icariine content is 31wt%, and the rate of recovery is 97.1%;
3) with POROS R
2(PE Biosystems company produce, component is a polystyrene) inverted medium is with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 40v% aqueous ethanolic solution balance; With step 2) icariine that obtains just separated product 440mg fully dissolve the back with 8ml 40% aqueous ethanolic solution and go up sample, after the abundant drip washing of 40v% aqueous ethanolic solution, with 45～80v% aqueous ethanolic solution gradient elution; UV270nm detects, and collects the 3rd elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 146.7mg again, measures wherein through HPLC that icariine content is 93wt%, and the rate of recovery is 100%;
4) the 146.7mg icariine that step 3) is obtained with the aqueous ethanolic solution of 40v% again separated product carry out recrystallization, obtain the pure product 124.6mg of icariine, purity is 98.5%, the rate of recovery is 90%.
Embodiment 6, be raw material, separate the pure product of icariine that the preparation of purifying contains icariine 97.5% with the over-ground part of the Da Hua Herba Epimedii grass that contains icariine 0.47%
1) after 10g contains the Da Hua Herba Epimedii grassland material leaf pulverizing of Herba Epimedii glucoside 0.47%, put into the 500ml matrass, as same quadrat method among the embodiment 1, the aqueous acetone solution refluxing extraction with 150ml 50% refluxed 1 hour, and refluxing extraction is 2 times altogether; Filter, with the aqueous acetone solution washing filter residue several of same concentration; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 1.8g, and the icariine content of measuring wherein through HPLC is 2.58wt%, and the rate of recovery is 99%;
2) HP-30 (MIT produces, and component is a polystyrene) is placed the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; With this chromatography column of 20v% isopropanol water solution balance; The icariine crude product 1.8g that step 1) is obtained fully dissolves upward sample of back with 20ml 20v% Virahol, after the abundant drip washing of 20v% isopropanol water solution, with 45v% isopropanol water solution wash-out; UV270nm detects, and collects second elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 143.6mg just, measures wherein through HPLC that icariine content is 32wt%, and the rate of recovery is 99%;
3) with TSK gel Octadecyl-NPR (TOSOH company produce, component is a polyacrylic ester) medium with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 25v% isopropanol water solution balance; With step 2) icariine that obtains just separated product 140mg fully dissolve the back with 5ml 25v% isopropanol water solution and go up sample, with the abundant drip washing chromatography column of 25v% isopropanol water solution, with 30～60v% isopropanol water solution gradient elution; UV270nm detects, and collects the 3rd elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 49.3mg again, measures wherein through HPLC that icariine content is 89wt%, and the rate of recovery is 98%:
4) the 49.3mg icariine that step 3) is obtained with the aqueous ethanolic solution of 45v% again separated product carry out recrystallization, obtain the pure product 42.3mg of icariine, purity is 97.5%, the rate of recovery is 94%.
Embodiment 7, be raw material, separate the pure product of icariine that the preparation of purifying contains icariine 97.5% with the over-ground part of the Epimedium wushanense grass that contains icariine 0.51%
1) after the Epimedium wushanense grassland material leaf that 10g is contained icariine 0.51% is pulverized, put into the 500ml matrass, as same quadrat method among the embodiment 1, with 40ml 50v% isopropanol water solution refluxing extraction, refluxed 1 hour at every turn, refluxing extraction is 2 times altogether; Filter, with the isopropanol water solution washing filter residue several of same concentration; Merging filtrate and washings, concentrating under reduced pressure obtains icariine crude product 1.96g, and the icariine content of measuring wherein through HPLC is 2.60wt%, and the rate of recovery is 100%;
2) S-8 resin (Chemical Plant of Nankai Univ. produces, and component is a polystyrene) is placed the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; With the resin after this activation with pack into the chromatography column of 15 * 200mm of settling methods; This chromatography column of aqueous ethanolic solution balance with 35v%; The icariine crude product 1.9g that step 1) is obtained fully dissolves the back with the aqueous ethanolic solution of 20ml 35v% and goes up sample, with the abundant drip washing chromatography column of aqueous ethanolic solution of 35v%, with the aqueous ethanolic solution wash-out of 65v%; UV270nm detects, and collects second elution peak; The concentrating under reduced pressure elutriant obtains icariine separated product 163mg just, measures wherein through HPLC that icariine content is 30wt%, and the rate of recovery is 99%;
3) with C
18Inverted medium is with pack into the chromatography column of 16 * 100mm of settling methods, with this chromatography column of 35v% methanol aqueous solution balance; With step 2) icariine that obtains just separated product 160mg fully dissolve the back with 10ml 35% methanol aqueous solution and go up sample, after the abundant drip washing of 35v% methanol aqueous solution, with 40～80v% methanol aqueous solution gradient elution; UV270nm detects, and collects the 3rd elution peak; Elutriant that concentrating under reduced pressure is received obtains icariine separated product 50.8mg again, measures wherein through HPLC that icariine content is 85wt%, and the rate of recovery is 90%;
4) the 50.8mg icariine that step 3) is obtained with the methanol aqueous solution of 45v% again separated product carry out recrystallization, obtain the pure product 40.3mg of icariine, purity is 97.5%, the rate of recovery is 91%.
The organic solvent of using in the present embodiment recycles after can reclaiming once more.
Claims (5)
1. method of from barrenwort, extracting icariine, its processing step is as follows:
1) lixiviate icariine from barrenwort: after the raw material leaf pulverized, with the aqueous solution (I) refluxing extraction of the organic solvent of 15～20 times of amounts of its weight; Filter then, with the aqueous solution (I) the washing filter residue several of organic solvent; Merging filtrate and washings, concentrating under reduced pressure obtains the icariine crude product; The content of organic solvent is 50～100v% in the aqueous solution of described organic solvent (I); Described organic solvent comprises methyl alcohol, ethanol, Virahol, propyl carbinol, acetone, methyl acetate, ethyl acetate;
2) just separate icariine: the polarity or the low-pole macro-reticular adsorbing resin of non-ionic type are placed the ethanol soaked overnight, use the drip washing successively of ethanol, deionized water then, so that the resin activation; Resin after this activation is adorned post with settling methods; As moving phase, the balance chromatography column is gone up sample with the icariine crude product that step 1) obtains with the aqueous solution (II) the dissolving back of organic solvent with the aqueous solution (II) of organic solvent; The aqueous solution (III) wash-out with organic solvent; Use the ultraviolet monitoring elution process, detect, collect second elution peak at 270nm; The elutriant that concentrating under reduced pressure is collected obtains icariine separated product just; The content of organic solvent is 5～25v% in the aqueous solution of described organic solvent (II); The content of organic solvent is 45～70v% in the aqueous solution of described organic solvent (III); Described organic solvent comprises methyl alcohol, ethanol, Virahol and acetone;
3) the first separated product of purification icariine: will be the inverted medium of matrix with silica gel or be that the inverted medium of matrix is adorned post with settling methods with the polymkeric substance; As moving phase, the balance chromatography column is with step 2 with the aqueous solution (IV) of organic solvent) icariine that obtains just separated product go up sample with the aqueous solution (IV) the dissolving back of organic solvent; With the aqueous solution (V) of organic solvent as elutriant, gradient elution; Use the ultraviolet monitoring elution process, detect, collect the 3rd elution peak at 270nm; The elutriant that concentrating under reduced pressure is collected obtains icariine separated product again; The content of organic solvent is 35～55v% in the aqueous solution of described organic solvent (IV); The content of organic solvent is 40～80v% in the aqueous solution of described organic solvent (V); Described organic solvent comprises methyl alcohol, ethanol and Virahol;
4) recrystallization: the icariine that step 3) is obtained with the methyl alcohol of 35～45v% or ethanol separated product again carries out recrystallization, obtains the pure product of icariine.
2. by the described method of extracting icariine from barrenwort of claim 1, it is characterized in that: the barrenwort of described step 1) comprises Da Ye Herba Epimedii, lobus cardiacus Herba Epimedii, arrow leaf Herba Epimedii, pubescence Herba Epimedii, Herba Epimedii, Da Hua Herba Epimedii and Epimedium wushanense.
3. by the described method of extracting icariine from barrenwort of claim 1, it is characterized in that: the number of times of the refluxing extraction in the described step 1) is 1～4 time; The time of described each refluxing extraction is 0.5～2 hour.
4. by the described method of from barrenwort, extracting icariine of claim 1, it is characterized in that: the resin described step 2) comprises polystyrene, polyacrylamide, polymethacrylate, acrylamide and cinnamic multipolymer, or methacrylic ester and cinnamic multipolymer.
5. by the described method of extracting icariine from barrenwort of claim 1, it is characterized in that: in the described step 3) is that the inverted medium of matrix is commercially available C with silica gel
8And C
18Inverted medium; In the described step 3) is that the inverted medium of matrix comprises that with polystyrene, polyacrylic ester, styrene diethylene benzene copoly mer, alkyl-modified polystyrene alcohol be the inverted medium of matrix with the polymkeric substance.
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| WO2008089669A1 (en) * | 2007-01-23 | 2008-07-31 | Tongjitang Chinese Medicines Company | A novel combination of epimedium-derived-flavonoids-fractions for prevention of steroid-induced osteonecrosis |
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|---|---|---|---|---|
| CN1295224C (en) * | 2004-11-11 | 2007-01-17 | 中国药科大学 | Process for preparing epimedium total flavone for injection |
| CN101302548B (en) * | 2007-05-09 | 2011-04-13 | 北京珅奥基医药科技有限公司 | Preparation of icaritin |
| CN101607976B (en) * | 2008-06-19 | 2013-09-04 | 贵州省中国科学院天然产物化学重点实验室 | Method for preparing icariin |
| CN101845067B (en) * | 2010-05-21 | 2012-06-27 | 甘肃亚兰特种药材饮片生产有限公司 | Preparation method of high-content icariin |
| CN102807595A (en) * | 2011-06-02 | 2012-12-05 | 汉中天然谷生物科技有限公司 | Technique for preparing high-content (98 percent) icariin from Epimedium plant |
| CN103910772B (en) * | 2014-04-18 | 2015-11-18 | 成都合盛生物技术有限公司 | A kind of method simultaneously extracting icarin and the precious leaves of pulse plants glycosides I, II from Herba Epimedii |
| CN104059116A (en) * | 2014-04-30 | 2014-09-24 | 西安岳达植物科技有限公司 | Method for separating icariine from herba epimedii |
| CN111560409B (en) * | 2020-05-08 | 2023-04-28 | 陕西嘉禾生物科技股份有限公司 | Preparation method of icariin |
| CN112725396A (en) * | 2020-09-22 | 2021-04-30 | 北京岳达生物科技有限公司 | Method for preparing icariside II through enzyme catalysis |
| DE212021000189U1 (en) * | 2021-10-17 | 2022-01-19 | Nantong Taiying New Material Science and Technology Co., Ltd. | Enrichment tube for enriching the total flavonoids in Epimedium leaf extract |
-
2003
- 2003-04-11 CN CN 03121821 patent/CN1225470C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008089669A1 (en) * | 2007-01-23 | 2008-07-31 | Tongjitang Chinese Medicines Company | A novel combination of epimedium-derived-flavonoids-fractions for prevention of steroid-induced osteonecrosis |
| CN101641112B (en) * | 2007-01-23 | 2013-01-23 | 同济堂药业 | A novel combination of epimedium-derived-flavonoids-fractions for prevention of steroid-induced osteonecrosis |
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| Publication number | Publication date |
|---|---|
| CN1535976A (en) | 2004-10-13 |
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