CN105925635A - Production process of icariside II - Google Patents
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- CN105925635A CN105925635A CN201610289040.1A CN201610289040A CN105925635A CN 105925635 A CN105925635 A CN 105925635A CN 201610289040 A CN201610289040 A CN 201610289040A CN 105925635 A CN105925635 A CN 105925635A
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Abstract
The invention discloses a production process of icariside II, wherein the production process comprises the following steps: firstly, extracting an icariin monomer from herba epimedii; and then, enzymatically hydrolyzing the icariin monomer by virtue of glycosyl hydrolase which is high in catalytic activity, so that the icariside II is obtained. The production process disclosed by the invention has the advantages of being efficient and gentle in reaction, low in cost, high in product purity and applicable to industrial production.
Description
Technical field
The present invention relates to the production technology of a kind of medicine, particularly to the production of a kind of icariside II
Technique.
Background technology
Erection disturbance (ED) is male's common disease, adds up according to World Health Organization, and the whole world suffers from ED
Male account for 10%, in man, the ratio of ED patient then reaches more than 1/4.China 3 hundred million one-tenth
In year male, there are about ED patient people more than 100,000,000, how to be caused by psychological or organic reason, and
Increasingly rejuvenation, has had a strong impact on man and wife physically and mentally healthy.
Anti-ED medicine can significantly improve the erection problem of ED patient.According to statistics, anti-ED in 2012
First-line drug world market terminal scale reaches 4,500,000,000 dollars.Adult male ED patient populations is huge in China,
Market potential capacity is in tens billion of units.The development and application of these anti-ED medicines, creates good
Society and economic benefits.
Current anti-ED drug main to have three big core product--sldenafil, tadalafil, Vardenafils,
They broadly fall into 5 type phosphodiesterase (PDE5) inhibitor, can improve cGMP concentration under sexual stimulus
And strengthen erection function.But owing to these oral selectivity PDE5 inhibitor are disposable strong only
Lax corpus cavernosal smooth muscle and promote erection, cure the symptoms, not the disease, the pairs such as hypotension can be caused simultaneously
Effect, the particularly safety to there being cardiovascular patient is too controversial.Therefore, exploitation safety,
The anti-ED new drug for the treatment of both the principal and secondary aspects of a disease becomes a kind of inevitable requirement.
Herba Epimedii is Berberidaceae Epimedium herbaceous plant, and it has 2000 so far as medea application
History for many years.The effective ingredient icariin monomer tool of the clinical observation display Herba Epimedii of BJ Univ Hospital
There is significant anti-ED effect, blood pressure is had no significant effect simultaneously.Zoopery shows its toxicity very
Low (50% icariin is to rat maximum toleration > 5g/kg).Icariin metabolic derivative in vivo
Icariside II has more significant biological function.Molecular studies show, icariside II list
Body is not only the specific inhibitor of PDE5, has some biologys of the effect that sldenafil does not has simultaneously
Should, as promoted the regeneration of penile nerve, improve expression and the endothelium of cavernous nerves of penis nNOS
Cell eNOS, promotes testosterone cell proliferation etc..Therefore, icariside II is expected to become new
Medicine research preparation, for treating and preventing novel drugs and the health product of erection disturbance, overcomes Xi Dina
The non-drawback waiting medicine " to cure the symptoms, not the disease ", wide market.
Research about icariin and icariside II the most both at home and abroad is concentrated mainly on pharmacology and merit
Research in terms of Neng, except in addition to the effect in terms of anti-ED, at the aspect such as defying age, antioxidation
Function is found the most in succession, but the biological manufacture view about icariside II is studied less, with state
It is inside main, is still within the starting stage.External in default of the resource in terms of Chinese medicine, research is compared
Limited.
The Jin Fengxie seminar of Dalian Polytechnic University takes the lead in developing the icariin enzyme of a kind of fungus, can
Icariin is degraded into low glycosyl glycosides or aglycon, then utilizes macroporous adsorbent resin SP207 to separate
Each component, product purity can reach 99%, but there is the technology such as stirring due to fungus liquid fermentation difficult
Point, fails to realize industrialized production.For overcoming this difficult problem, this seminar is from the soil of cultivated ginseng
In filtered out the bacterial isolates Rhodanobacter sp.GS3054 that can hydrolyze icariin glycosyl, and
Have studied its fermentation condition and enzyme reaction parameter, the ability relatively fungal enzyme of hydrolysis glycosyl has promoted.By
This, the fungus and the antibacterial that possess hydrolysis glycosyl ability are the most all developed, but are dividing owing to lacking
, there is the problems such as hydrolysis efficiency is low, it is impossible to pass through genetic engineering in the understanding to glycosyl hydrolase of the sub-level
Hydrolytic enzyme expression in antibacterial or fungus and activity are greatly improved.Directly find highly active glycosyl
Hydrolytic enzyme, and it is directly used in biocatalytic reaction hydrolytic enzyme gradually by gene engineering method great expression
Become new study hotspot.
Summary of the invention
It is an object of the invention to provide the production technology of a kind of icariside II, there is reaction efficiently,
Gentleness, low cost, product purity is high, the advantage being suitable for industrialized production.
The technical solution adopted for the present invention to solve the technical problems is:
The production technology of a kind of icariside II, first extracts icariin monomer, then from Herba Epimedii
Glycosyl hydrolase enzymolysis icariin monomer is utilized to obtain icariside II.
As preferably, the processing step extracting icariin monomer from Herba Epimedii is as follows: after Herba Epimedii is pulverized,
Add mass concentration 45-85% alcohol heating reflux 1-4h and obtain Herba Epimedii crude extract；Herba Epimedii crude extract is used
Water and acetic acid ethyl dissolution are also layered, and aqueous phase is extracted with ethyl acetate 3-5 time, combined ethyl acetate phase,
Washing with saturated aqueous common salt, anhydrous sodium sulfate is dried, and is spin-dried for, and obtains dried object；Dried object mixing is molten
Agent is heated to reflux 1-4h, filters, and filtering residue is heated to reflux 1-4h with mixed solvent again, filters, and merges filter
Liquid, dried icariin monomer.
Present invention improves over icariin monomer extraction process, its advantage is: directly use mixed solvent
Repeated crystallization method, lower than traditional chromatographic column or macroporous adsorbent resin separation method cost (main
If solvent load is few, no-arbitrary pricing expense, equipment requirements low), with short production cycle, it is simple to the big rule of factory
Mould produces.
As preferably, Herba Epimedii is crushed to 20-80 mesh, and 100g Herba Epimedii ground product adds 300-800mL mass
Concentration 45-85% ethanol.
As preferably, described mixed solvent is water with methanol according to the mixture of the volume ratio of 1:2-3.
As preferably, described glycosyl hydrolase is prepared by the following method and obtains: first with coccus containing Fructus Vitis viniferae
Cultivate to produce glucosan on the solid medium of sugar, then with this glucosan preparation fluid medium with training
Supporting zymogenic bacteria, leach the thalline of zymogenic bacteria, thalline is dissolved in Acetic acid-sodium acetate buffer immersion 3-5h,
Centrifugal, take supernatant, after lyophilization, obtain the glycosyl hydrolase of high catalytic activity.
First cultivate to produce glucosan in the culture medium containing glucose with coccus specifically comprises the processes of:
Solid culture based formulas is: glucose 15g, peptone 10g, Semen Maydis pulp 35g, agar 15g,
H2O 1000mL, dipotassium hydrogen phosphate-potassium phosphate buffer regulation pH to 5.5～7.5；Temperature
30 DEG C, cultivate 24h.Solid medium after cultivation according to every gram of solid medium with 3～8mL
The amount of water soaks 3～5h, is centrifuged, discards solid, obtain glucosan clear liquid.The phosphoric acid hydrogen two used
Potassium-potassium phosphate buffer pH is 7.8-8.0.
Glucosan preparation fluid medium is to cultivate zymogenic bacteria specifically comprises the processes of:
Liquid culture based formulas is: glucosan clear liquid 15g, peptone 10g, H2O 1000mL, phosphorus
Acid hydrogen dipotassium-potassium phosphate buffer regulation pH 5.5～7.5；Cultivate 4～5 days at 30 DEG C, filter
Going out thalline, thalline is dissolved in the Acetic acid-sodium acetate buffer of pH value 5.5 and soaks 3-5h, and every gram of thalline makes
With the Acetic acid-sodium acetate buffer of 3-8mLpH value 5.5, centrifugal (rotating speed 3000-8000rmp), take
Supernatant, obtains the glycosyl hydrolase (powder glucanase) of high catalytic activity after lyophilization.
Can be screened by the above-mentioned special process of the present invention and obtain a kind of high catalytic activity, reaction condition temperature
The glucanase (glycosyl hydrolase of high catalytic activity) of sum.This kind of glucanase is for icariin
Selectivity is the highest, and transformation efficiency is close to 100%.
As preferably, described coccus is Rhodococcus fascians or bright string coccus.
As preferably, described zymogenic bacteria is that penicillium sp, aspergillosis, wheel be mould, aspergillus niger or bifidus.
As preferably, enzymolysis process is: 100mg icariin monomer is suspended in the phosphoric acid hydrogen of pH=5.5-7.0
Dipotassium-potassium phosphate buffer obtains cumulative volume 10-20mL reaction system, adds in reaction system
The solubilizing agent of reaction system volume 4-6% and glycosyl hydrolase, be stirred overnight, ethyl acetate extraction 3-5
Secondary, merge organic facies, washing, saturated aqueous common salt washs, and anhydrous sodium sulfate is dried, and concentrates rear pump or output pump
Drain to obtain product.
As preferably, described solubilizing agent is Tween 80, glycerol or dimethyl sulfoxide.
As preferably, described glycosyl hydrolase enzyme dosage is the 10-20% of icariin monomer weight.
The invention has the beneficial effects as follows:
There is the problems such as purity is relatively low, impurity content is higher, the present invention in current Extraction Technology of Icariin from Herba Epimedii
Optimize the extraction process of icariin, by the environmental protection reagent such as widely used ethanol extraction, fall
Low environmental pollution, and by increasing re-crystallization step, improve the purity of product, reach 98.9%.Excessive
The sheep leaves of pulse plants time glycosides II and the difference only one of which glycosyl of icariin, separate both materials and exist certain
The problem that difficulty and cost increase, the present invention utilizes the glycosyl hydrolase of high catalysis, in 5kg rank
In pilot scale, the conversion ratio of reaction can be reached more than 99%, simplify the purification step of icariside II
Suddenly.
Accompanying drawing explanation
Fig. 1 is the liquid chromatogram of product icariside II of the present invention.
Detailed description of the invention
Below by specific embodiment, technical scheme is described in further detail.
In the present invention, if not refering in particular to, the raw material used and equipment etc. are all commercially available or this
Field is commonly used.Method in following embodiment, if no special instructions, is the routine side of this area
Method.
Total embodiment:
A kind of production technology of icariside II:
(1) from Herba Epimedii, icariin monomer is first extracted: from Herba Epimedii, extract icariin monomer
Processing step is as follows: Herba Epimedii is crushed to 20-80 mesh, and 100g Herba Epimedii ground product adds 300-800mL
Mass concentration 45-85% ethanol, is heated to reflux 1-4h and obtains Herba Epimedii crude extract；Herba Epimedii crude extract is used
Water and acetic acid ethyl dissolution are also layered, and aqueous phase is extracted with ethyl acetate 3-5 time, combined ethyl acetate phase,
Washing with saturated aqueous common salt, anhydrous sodium sulfate is dried, and is spin-dried for, and obtains dried object；Dried object mixing is molten
Agent is heated to reflux 1-4h, filters, and filtering residue is heated to reflux 1-4h with mixed solvent again, filters, and merges filter
Liquid, dried icariin monomer.Mixed solvent is water with methanol according to volume ratio mixed of 1:2-3
Compound.
(2) the glycosyl hydrolase enzymolysis icariin monomer then utilizing high catalytic activity obtains icariside
II: 100mg icariin monomer is suspended in the dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffering of pH=5.5-7.0
Liquid obtains cumulative volume 10-20mL reaction system, in reaction system, adds reaction system volume 4-6%'s
Solubilizing agent (Tween 80, glycerol or dimethyl sulfoxide) and the glycosyl hydrolase of high catalytic activity are (described
The 10-20% that glycosyl hydrolase enzyme dosage is icariin monomer weight of high catalytic activity), it is stirred overnight,
Ethyl acetate extracts 3-5 time, merges organic facies, washing, and saturated aqueous common salt washs, anhydrous sodium sulfate
It is dried, concentrates rear pump or output pump and drain to obtain product.
The glycosyl hydrolase of described high catalytic activity is prepared by the following method and obtains: first with coccus (Rhodococcus fascians
Or bright string coccus) on the solid medium containing glucose, cultivate to produce glucosan, then use this Portugal
(penicillium sp, aspergillosis, wheel be mould, aspergillus niger or bifid breast to cultivate zymogenic bacteria for polysaccharide preparation fluid medium
Bacillus), leach the thalline of zymogenic bacteria, thalline is dissolved in Acetic acid-sodium acetate buffer immersion 3-5h, from
The heart, takes supernatant, obtains the glycosyl hydrolase of high catalytic activity after lyophilization.
The coccus, the zymogenic bacteria that use in the present invention are commercially available prod.
It is embodied as technique:
Icariin monomer is prepared from Herba Epimedii
Embodiment 1:
Herba Epimedii grass (40-60 mesh) 100g, 75% ethanol 400ml is heated to reflux 1h, obtains 15.5g excessive
Sheep leaves of pulse plants crude extract (HPLC Icariin content 10%), molten with 150ml water and 150ml ethyl acetate
Solving and be layered, aqueous phase extracts 3 times by 150ml ethyl acetate every time, combined ethyl acetate phase, with full
And brine It, anhydrous sodium sulfate is dried, and is spin-dried for obtaining 8.5g Tan solid.With water: methanol=1:2
Mixed solvent 100ml be heated to reflux 1h, filter, filtering residue again with 100ml mixed solvent reflux 1h,
Merge the mother solution of twice, be spin-dried for, dry to obtain 1.49g light yellow solid (HPLC Icariin content
98.5%), yield 96.1%.
Embodiment 2:
Herba Epimedii grass (20-40 mesh) 100g, 75% ethanol 500ml is heated to reflux 1h, obtains 14.5g excessive
Sheep leaves of pulse plants crude extract (HPLC Icariin content 10%), molten with 150ml water and 150ml ethyl acetate
Solving and be layered, aqueous phase extracts 3 times by 150ml ethyl acetate every time, combined ethyl acetate phase, with full
And brine It, anhydrous sodium sulfate is dried, and is spin-dried for obtaining 8.2g Tan solid.With water: methanol=1:2
Mixed solvent 100ml be heated to reflux 1h, filter, filtering residue again with 100ml mixed solvent reflux 1h,
Merge the mother solution of twice, be spin-dried for, dry to obtain 1.35g light yellow solid (HPLC Icariin content
98.3%), yield 93.1%.
Embodiment 3:
Herba Epimedii grass (60-80 mesh) 100g, 75% ethanol 600ml is heated to reflux 1h, obtains 15.2g excessive
Sheep leaves of pulse plants crude extract (HPLC Icariin content 10%), molten with 150ml water and 150ml ethyl acetate
Solving and be layered, aqueous phase extracts 3 times by 150ml ethyl acetate every time, combined ethyl acetate phase, with full
And brine It, anhydrous sodium sulfate is dried, and is spin-dried for obtaining 8.3g Tan solid.With water: methanol=1:2
Mixed solvent 100ml be heated to reflux 1h, filter, filtering residue again with 100ml mixed solvent reflux 1h,
Merge the mother solution of twice, be spin-dried for, dry to obtain 1.44g light yellow solid (HPLC Icariin content
98.5%), yield 94.7%.
The glycosyl hydrolase screening of high catalytic activity:
Embodiment 4:
First with rhodococcus erythropolis (commercially available, to be purchased from Chinese microorganism strain inquiry net) containing Fructus Vitis viniferae
Cultivating to produce glucosan on the solid medium of sugar, solid culture based formulas is: glucose 15g,
Peptone 10g, Semen Maydis pulp 35g, agar 15g, H2O 1000mL, dipotassium hydrogen phosphate-di(2-ethylhexyl)phosphate
Hydrogen potassium buffer regulation pH 6.0.Incubator 30 DEG C, cultivates 24h.Solid culture after cultivation
Base soaks 3～5h according to every gram of solid medium amount of 5mL water, with centrifuge, discards solid
Body, obtains glucosan clear liquid.Again with this glucosan preparation fluid medium to cultivate Penicillium griseofulvum (city
Sell, be purchased from Chinese microorganism strain inquiry net), liquid culture based formulas is: glucosan clear liquid 15g,
Peptone 10g, H2O 1000mL, dipotassium hydrogen phosphate-potassium phosphate buffer regulation pH6.0；?
Cultivating 4～5 days at 30 DEG C, leach thalline, thalline is dissolved in the Acetic acid-sodium acetate buffer of pH value 5.5
Soaking 3-5h, every gram of thalline uses the Acetic acid-sodium acetate buffer of 5mLpH value 5.5, is centrifuged and (turns
Speed 3000-8000rmp), take supernatant, after lyophilization, obtain the glycosyl hydrolase (powder of high catalytic activity
Powder glucanase).
Embodiment 5:
The present embodiment difference from Example 4 is: first with Rhodococcus ruber (commercially available, be purchased from China micro-
Biological inoculum inquiry net) cultivate to produce glucosan on the solid medium containing glucose.Use again
This glucosan preparation fluid medium with cultivate Aspergillus flavus (commercially available, be purchased from Chinese microorganism strain
Inquiry net).
The other the same as in Example 4.
Embodiment 6:
The present embodiment difference from Example 4 is: first with band Rhodococcus fascians (commercially available, be purchased from China
Microorganism fungus kind inquiry net) cultivate to produce glucosan on the solid medium containing glucose.Again
With this glucosan preparation fluid medium with cultivate penicillium (commercially available, be purchased from Chinese microorganism strain
Inquiry net).
The glycosyl hydrolase enzymolysis icariin monomer utilizing high catalytic activity obtains icariside II:
Embodiment 7:
100mg icariin is suspended in the dipotassium hydrogen phosphate-potassium phosphate buffer of pH=6.8 totally
Long-pending 10mL reaction system, adds the Tween 80 of reaction system volume 5%, 10mg Portugal in reaction system
Dextranase (glycosyl hydrolase of high catalytic activity, the i.e. enzyme of embodiment 4-6 screening), is stirred overnight.
Ethyl acetate 30mL extracts in three times, merges organic facies, and 10mL washes, and saturated aqueous common salt washs,
Anhydrous sodium sulfate is dried, and concentrates rear pump or output pump and drains to obtain solid 75.3mg, productivity 99.1%.HPLC is pure
Degree is 99.4%.
Embodiment 8:
100mg icariin is suspended in the dipotassium hydrogen phosphate-potassium phosphate buffer of pH=7.0 totally
Long-pending 10mL reaction system, adds the glycerol of reaction system volume 5%, 10mg Portugal in reaction system
Dextranase (glycosyl hydrolase of high catalytic activity, the i.e. enzyme of embodiment 4-6 screening), is stirred overnight.
Ethyl acetate 30mL extracts in three times, merges organic facies, and 10mL washes, and saturated aqueous common salt washs,
Anhydrous sodium sulfate is dried, and concentrates rear pump or output pump and drains to obtain solid 73.8mg, productivity 97.1%.HPLC is pure
Degree is 98.5%.
Embodiment 9:
100mg icariin is suspended in the dipotassium hydrogen phosphate-potassium phosphate buffer of pH=6.5 totally
Long-pending 10mL reaction system, adds the dimethyl sulfoxide of reaction system volume 5% in reaction system,
10mg glucanase (glycosyl hydrolase of high catalytic activity, the i.e. enzyme of embodiment 4-6 screening), stirs
Mix overnight.Ethyl acetate 30mL extracts in three times, merges organic facies, and 10mL washes, saturated common salt
Water washs, and anhydrous sodium sulfate is dried, and concentrates rear pump or output pump and drains to obtain solid 74.5mg, productivity 98.0%.
HPLC purity is 98.5%.
The product of the present invention confirms as Herba Epimedii after liquid chromatograph (Fig. 1) detection contrasts with standard substance
Glycosides II.
Embodiment described above is the one preferably scheme of the present invention, not appoints the present invention
What pro forma restriction, also has other on the premise of without departing from the technical scheme described in claim
Variant and remodeling.
Claims (10)
1. the production technology of an icariside II, it is characterised in that: from Herba Epimedii, first extract excessive sheep
Icariin monomer, then utilizes glycosyl hydrolase enzymolysis icariin monomer to obtain icariside II.
The production technology of a kind of icariside II the most according to claim 1, it is characterised in that: from
The processing step extracting icariin monomer in Herba Epimedii is as follows: after Herba Epimedii is pulverized, add mass concentration
45-85% alcohol heating reflux 1-4h obtains Herba Epimedii crude extract；Herba Epimedii crude extract water and ethyl acetate
Dissolving and be layered, aqueous phase is extracted with ethyl acetate 3-5 time, combined ethyl acetate phase, uses saturated common salt
Water washs, and anhydrous sodium sulfate is dried, and is spin-dried for, and obtains dried object；Dried object mixed solvent is heated to reflux
1-4h, filters, and filtering residue is heated to reflux 1-4h with mixed solvent again, filters, merging filtrate, after drying
Obtain icariin monomer.
The production technology of a kind of icariside II the most according to claim 2, it is characterised in that: excessive
The sheep leaves of pulse plants is crushed to 20-80 mesh, and 100g Herba Epimedii ground product adds 300-800mL mass concentration 45-85%
Ethanol.
The production technology of a kind of icariside II the most according to claim 2, it is characterised in that: institute
Stating mixed solvent is water with methanol according to the mixture of the volume ratio of 1:2-3.
The production technology of a kind of icariside II the most according to claim 1 and 2, it is characterised in that:
Described glycosyl hydrolase is prepared by the following method and obtains: first train at the solid containing glucose with coccus
Support and on base, cultivate to produce glucosan, then with this glucosan preparation fluid medium to cultivate zymogenic bacteria,
Leaching the thalline of zymogenic bacteria, thalline is dissolved in Acetic acid-sodium acetate buffer immersion 3-5h, centrifugal, takes
Clear liquid, obtains the glycosyl hydrolase of high catalytic activity after lyophilization.
The production technology of a kind of icariside II the most according to claim 5, it is characterised in that: institute
Stating coccus is Rhodococcus fascians or bright string coccus.
The production technology of a kind of icariside II the most according to claim 5, it is characterised in that: institute
State zymogenic bacteria be that penicillium sp, aspergillosis, wheel be mould, aspergillus niger or bifidus.
The production technology of a kind of icariside II the most according to claim 1, it is characterised in that: enzyme
Solution technique is: 100mg icariin monomer is suspended in the dipotassium hydrogen phosphate-biphosphate of pH=5.5-7.0
Potassium buffer obtains cumulative volume 10-20mL reaction system, in reaction system, adds reaction system volume
The solubilizing agent of 4-6% and glycosyl hydrolase, be stirred overnight, and ethyl acetate extracts 3-5 time, merges organic
Phase, washing, saturated aqueous common salt washs, and anhydrous sodium sulfate is dried, and concentrates rear pump or output pump and drains to obtain product.
The production technology of a kind of icariside II the most according to claim 1, it is characterised in that: institute
Stating solubilizing agent is Tween 80, glycerol or dimethyl sulfoxide.
The production technology of a kind of icariside II the most according to claim 1, it is characterised in that:
Described glycosyl hydrolase enzyme dosage is the 10-20% of icariin monomer weight.
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| CN108392487A (en) * | 2018-02-24 | 2018-08-14 | 苏州广奥医药开发有限公司 | A kind of application of icariside II or its pharmaceutical acceptable carrier in erectile dysfunction |
| CN110699263A (en) * | 2019-10-29 | 2020-01-17 | 浙江工业大学 | Aspergillus niger YH-6 and application thereof in improving content of icaritin in epimedium |
| CN112063672A (en) * | 2020-09-23 | 2020-12-11 | 浙大宁波理工学院 | Process for preparing icariside II by using exoglucanase |
| CN112080538A (en) * | 2020-09-21 | 2020-12-15 | 浙大宁波理工学院 | Method for preparing icariside II based on enzyme catalysis |
| CN112176008A (en) * | 2020-10-13 | 2021-01-05 | 烟台大学 | Enzymolysis method for efficiently and quickly preparing icariside II |
| 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 |
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| CN107964555B (en) * | 2018-01-05 | 2021-07-20 | 苏州广奥医药开发有限公司 | Method for biological enrichment and production of icariside II |
| CN108392487A (en) * | 2018-02-24 | 2018-08-14 | 苏州广奥医药开发有限公司 | A kind of application of icariside II or its pharmaceutical acceptable carrier in erectile dysfunction |
| CN108392487B (en) * | 2018-02-24 | 2023-09-01 | 北京东方百奥医药开发有限公司 | Application of icariside II or pharmaceutically acceptable carrier thereof in erectile dysfunction |
| CN110699263A (en) * | 2019-10-29 | 2020-01-17 | 浙江工业大学 | Aspergillus niger YH-6 and application thereof in improving content of icaritin in epimedium |
| CN110699263B (en) * | 2019-10-29 | 2021-05-11 | 浙江工业大学 | Aspergillus niger YH-6 and application thereof in improving content of icaritin in epimedium |
| CN112080538A (en) * | 2020-09-21 | 2020-12-15 | 浙大宁波理工学院 | Method for preparing icariside II based on enzyme catalysis |
| CN112080538B (en) * | 2020-09-21 | 2022-03-25 | 浙大宁波理工学院 | Method for preparing icariside II based on enzyme catalysis |
| 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 |
| CN112063672A (en) * | 2020-09-23 | 2020-12-11 | 浙大宁波理工学院 | Process for preparing icariside II by using exoglucanase |
| CN112176008A (en) * | 2020-10-13 | 2021-01-05 | 烟台大学 | Enzymolysis method for efficiently and quickly preparing icariside II |
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