CN106148449A - A kind of preparation method of icariside I - Google Patents
A kind of preparation method of icariside I Download PDFInfo
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- CN106148449A CN106148449A CN201510129279.8A CN201510129279A CN106148449A CN 106148449 A CN106148449 A CN 106148449A CN 201510129279 A CN201510129279 A CN 201510129279A CN 106148449 A CN106148449 A CN 106148449A
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- herba epimedii
- extract
- icariside
- enzyme digestion
- digestion reaction
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Abstract
The present invention provides the preparation method of a kind of icariside I, and the method comprises the following steps: first Herba Epimedii extract carries out under the effect of beta-glycosidase enzyme digestion reaction；Then enzyme digestion reaction product step a obtained filters；Filter cake ethyl alcohol recrystallization after filtering again twice, obtains described icariside I sterling.Icariside I in the present invention reaches 15% relative to Herba Epimedii extract yield, and the purity of icariside I reaches 99%.
Description
Technical field
The present invention relates to the preparation method of a kind of icariside I, belong to food, health product, change
Cosmetic and field of medicaments.
Background technology
Icariside I, has another name called Icariside I, is to extract to separate from Chinese crude drug Herba Epimedii
A kind of effective monomer arrived, shown in its structure such as following formula (I):
Icariside I, is the one in Herba Epimedii total flavones, has kidney-replenishing, bone and muscle strengthening, dispels
Effect of rheumatism.For kidney yang deficiency, impotence and seminal emission, muscles and bones flaccidity is soft, rheumatic arthralgia, numb inflexible
The symptoms such as contraction.Modern pharmacology finds that this type of material has an antitumor, antioxidation, anti-inflammation,
Protection cardio-cerebrovascular effect.
The preparation method of icariside I is disclosed in the patent document of CN101891782A.The party
Method is with Korean epimedium leaves as raw material, by alcohol reflux, adds concentrated hydrochloric acid mistake in crude extract
Filter, adds distilled water and separates out precipitation in filtrate, washing with alcohol precipitates, and is dried, and obtains Herba Epimedii
Glycosides I.Icariside I content in different Herba Epimedii kinds is the lowest, typically below 0.1%,
Main Flavonoids composition in major part epimedium herb is icariin.The method uses HCl treatment,
Its object is to the icariin in Herba Epimedii extract by concentrated hydrochloric acid hydrolysis is icariside
I.But icariin is under hydrochloric acid effect, and only small part is hydrolyzed to icariside I, the biggest portion
Branch thoroughly hydrolyzes and removes all glycosyls, generates 3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one.On the other hand, hydrochloric acid hydrolysis is used
Icariin, can destroy the isopentene group on flavone ring, causes hydrolysate impurities a lot.Therefore,
The method just can only can obtain icariside I sterling by chromatographic technique so that this technique is not suitable for
Industrialized production.
The extraction of icariside I is disclosed in the patent document of Publication No. CN101899077A
Method and application.In the method, first use alcohol reflux Folium Epimedii, then by extracting solution
Middle addition concentrated hydrochloric acid, heating and filtering, then separate out precipitation by filtrate adds distilled water, therefore pass through
The icariside I that the method obtains.But the shortcoming of the method lies also in and uses concentrated hydrochloric acid hydrolysis excessive
Sheep leaves of pulse plants extracting solution, can destroy the isopentene group on flavone ring, and the icariside I therefore obtained is miscellaneous
Matter is many, and purity is little.
It is thus desirable to a kind of method obtaining high-purity icariside I.
Summary of the invention
It is an object of the present invention to provide the preparation method of a kind of icariside I, by the party's legal system
The standby icariside I obtained has the advantage that conversion ratio is high, purity is high and impurity is few.
One aspect of the present invention provides the preparation method of a kind of icariside I, and the method includes following
Step:
First, by Herba Epimedii extract under the effect of beta-glycosidase, enzyme digestion reaction is carried out；
The most then enzyme digestion reaction product step a obtained filters；
Filter cake alcohol crystal after filtering the most again, obtains described icariside I.
Preferably, in the Herba Epimedii extract in described step a icariin mass content at least
15%.
Preferably, in described step a, it is 3.5-7.5 that Herba Epimedii extract is scattered in pH value
Buffer solution in, and enzyme digestion reaction temperature is 35-65 DEG C.
Preferably, described buffer solution is selected from the disodium hydrogen phosphate-phosphoric acid for pH value 5.0-6.0
Potassium dihydrogen, acetic acid-sodium acetate, phosphoric acid-sodium phosphate, citric acid-sodium citrate and disodium hydrogen phosphate-
One or more in citric acid solution.
Most preferably, described volume of buffer solution/L with the quality/Kg ratio of Herba Epimedii extract is
5-15:1。
Preferably, the enzyme digestion reaction time described in step a is 1-20 hour.
It is highly preferred that the described enzyme digestion reaction time is 10-20 hour.
Most preferably, the described enzyme digestion reaction time is 15-20 hour.
Preferably, the weight ratio of the Herba Epimedii extract in step a and beta-glycosidase is 1-50:1.
It is highly preferred that the weight ratio of Herba Epimedii extract and beta-glycosidase is 2-20:1 in step a.
Most preferably, in step a, the weight ratio of Herba Epimedii extract and beta-glycosidase is 5-10:1.
Preferably, in described step c, the filtration cakes torrefaction after first filtering, then carry out
Pulverize, then will pulverize after filter cake be scattered in ethanol backflow dissolve, filter, ethanol in filtrate
Volume/L is equivalent to pulverize 10-100 times of filter cake quality/Kg.
Preferably, in described step c, will crystal crude product, backflow in addition ethanol for the first time
Dissolving, filter, then filtrate cooling again crystallized, it is thick that the volume/L of refluxing ethanol is equivalent to crystallization
20-300 times of quality/Kg.
Preferably, in described step a, described Herba Epimedii extract is carried by alcohol solvent
Take Herba Epimedii Chinese crude drug to prepare.
Preferably, described alcohol solvent is ethanol water mixed solvent, and ethanol water mixed solvent extracts
The extract that Herba Epimedii Chinese crude drug obtains first passes around concentration, then passes through macroporous resin column absorption,
Carry out eluting with the ethanol water solvent that volumetric concentration is 30%-100% again, obtain for enzyme digestion reaction
Herba Epimedii extract.
The beneficial effects of the present invention is: 1. the present invention is with Herba Epimedii extract as raw material, will extract
Icariin in thing is optionally converted into icariside I, and conversion ratio reaches 70%, fully profit
With herb resource.2. the present invention uses the method for enzymolysis to remove the part glycosyl in icariin,
Reaction condition is gentle, and impurity is less, only just can get, by the mode of recrystallization, the excessive sheep that purity is 99%
The leaves of pulse plants time glycosides I, it is simple to industrialized great production.
Accompanying drawing explanation
Fig. 1 upper half figure represents the height of the total flavones in embodiment 1 step 1.1 ethanol extract
Effect liquid phase chromatogram figure.
Fig. 1 lower half figure represents total yellow in embodiment 1 step 1.2 ethanol extract after purification
The high-efficient liquid phase chromatogram of ketone.
Fig. 2 represents the high-efficient liquid phase chromatogram of icariside I sterling.
Fig. 3 is the mass spectrum [M-H] of icariside I sterling-。
Detailed description of the invention
Unless otherwise indicated, term " enzyme digestion reaction " herein refers to the water under enzyme effect
Solve reaction.
Unless otherwise indicated, " Herba Epimedii extract " of the terms refers to by ethanol water
The extract that solvent extraction Herba Epimedii obtains, containing the flavone including icariin in extract
Constituents and other compositions.
Unless otherwise indicated, term " bed volume " herein is filled out in referring to macroporous resin column
Fill the cumulative volume of thing.
Unless otherwise indicated, term " beta-glycosidase " herein be Japan Tian Ye company β-
Glycosidase, trade name Aromase H2.
Embodiment 1
1. the preparation of Herba Epimedii extract
1.1 extract
The present embodiment extracts by several times except stem Epimedium sagittatum leaf amounts to 400Kg, is rubbed broken, respectively
By 14 times, the ethanol water reflux, extract, of 10 times amount (v/w) 40% volumetric concentration 2 times, each 1
Hour.Reclaim ethanol to extracting solution without alcohol taste, detected in this extracting solution by high performance liquid chromatography
Total flavones, is shown in accompanying drawing 1 upper half figure.
1.2 purification by macroporous resin
Ethanol extraction step 1.1 obtained, concentrates after removing ethanol, uses purification by macroporous resin.
First with the distilled water eluting macroporous resin column of macroporous resin bed volume 8 times, the stream of distilled water eluting
Speed be 1 times of column volume/hour, after eluting terminates, discard eluent；
Macroporous resin column is washed with macroporous resin bed volume 6 times amount volumetric concentration 30% ethanol water
Bed, after eluting terminates, discards eluent；
With the macroporous resin column in the ethanol water solvent washing step 2 that volumetric concentration is 60%, ethanol water
Solvent volume is 4 times of bed volume, regulation ethanol water flow velocity be 1 times of bed volume/hour,
Collect eluent now.Reclaim ethanol and be concentrated into thick paste, being spray-dried, obtain Herba Epimedii and carry
Take thing and amount to 9.8Kg.Herba Epimedii extract after the purification by macroporous resin that above three-step approach obtains passes through
High performance liquid chromatography detects flavone component therein, sees Fig. 1 lower half figure.
Macroporous resin in the present embodiment is purchased from Hebei Bao En adsorbing material company limited, and model is
HPD-300 type.
High performance liquid chromatography (HPLC) condition: chromatographic column: Agela-C18 post；Flowing phase: second
Nitrile-water (wherein containing volume fraction is the trifluoroacetic acid of 0.0125%), gradient see table 1, washes
De-time 60min, flow velocity: 1.0mL/min, column temperature: 35 DEG C.UV-detector detection wavelength divides
Be not 254,272,320nm.
Table 1 solvent gradient elution table
Result shows: when at 3 minutes occur peak be solvent peak in addition to, 22 minutes, 24 minutes,
The peak that goes out of 46 minutes and 52 minutes is the peak of flavone component, the medicine detected by above method
The total flavones collection of illustrative plates of material, contrasts permissible with the collection of illustrative plates (HPLC-UV) of total flavones in Herba Epimedii extract
Finding out, after purification by macroporous resin, in Herba Epimedii extract, total flavones composition is effectively enriched with, its
In Herba Epimedii extract after purification relative to epimedium herb, wherein the mass yield of total flavones reaches
2.0～10.0%.
2. enzyme digestion reaction prepares icariside I
Weigh the above-mentioned Herba Epimedii extract 1.0Kg through purification by macroporous resin, join 15L
In 1.0mol/L pH=5.9 acetic acid-sodium acetate buffer solution, mixing speed is to make in the case of 100rpm
It dissolves.Until system temperature constant 60 DEG C of conditions time, add 0.1Kg beta-glycosidase stirring reaction.
After isothermal reaction 3 hours, stopping stirring, reactant liquor is cooled to 30 DEG C.
3. the purification of icariside I
After enzyme digestion reaction terminates, reactant liquor is centrifuged, and collects filter cake, and dried weight is 0.70Kg,
Add 35Kg ethanol after pulverizing, be heated to reflux stirring filtered while hot after 1 hour, filtrate slow cooling
After 25 DEG C, it is incubated 10 hours, filters, collect filter cake and be dried, obtaining icariside I once
Crystallized product 0.21Kg.16Kg ethanol is added after being pulverized by icariside I primary crystallization product,
Filter after being heated to reflux 1 hour, after filtrate slow cooling to 25 DEG C, be incubated 10 hours, filter, receive
Collection filter cake is also dried, and obtains icariside I recrystallized product and amounts to 0.12Kg, its liquid chromatograph and
As shown in Figures 2 and 3, Fig. 2 retention time at the peak of the local appearance of 6.037min is mass spectrum
The peak of icariside I, there is the peak that nucleocytoplasmic ratio is 529.4 in Fig. 3, represents [M-H]-, therefore lead to
Cross the icariside I purity showing to be obtained illustrated above by the inventive method higher.
Embodiment 2
1. enzyme digestion reaction prepares icariside I
Weigh commercial Herba Epimedii extract 1Kg, join 15L 0.8mol/L pH=6.0 phosphoric acid hydrogen two
In sodium-citric acid solution, mixing speed is to make it dissolve in the case of 100rpm.Treat system temperature
Spend constant when 55 DEG C of conditions, add the stirring reaction of 0.2Kg beta-glycosidase.Isothermal reaction 6 hours
After, stopping stirring, reactant liquor is cooled to 25～30 DEG C.
2. the purification of icariside I
Being centrifuged by above-mentioned reactant liquor, collect filter cake, dried weight is 0.86Kg, after pulverizing
Add 60.2Kg ethanol, be heated to reflux stirring filtered while hot after 1 hour, filtrate slow cooling to 25 DEG C
Rear insulation 10 hours, filters, and collects filter cake and is dried, and obtains icariside I primary crystallization and produces
Thing 0.26Kg.Add 20Kg ethanol after being pulverized by icariside I primary crystallization product, heat back
Filter after flowing 1 hour, after filtrate slow cooling to 25 DEG C, be incubated 10 hours, filter, collect filter cake
And be dried, obtain icariside I recrystallized product and amount to 0.15Kg.
Claims (10)
1. a preparation method for icariside I, the method comprises the following steps:
First, by Herba Epimedii extract under the effect of beta-glycosidase, enzyme digestion reaction is carried out；
The most then enzyme digestion reaction product step a obtained filters；
Filter cake alcohol crystal after filtering the most again, obtains described icariside I.
Method the most according to claim 1, it is characterised in that excessive sheep in described step a
Icariin mass content at least 15% in leaves of pulse plants extract.
Method the most according to claim 1, it is characterised in that in described step a,
Herba Epimedii extract is scattered in the buffer solution that pH value is 3.5-7.5, and enzyme digestion reaction temperature
For 35-65 DEG C；Preferably, described buffer solution selected from pH value 5.0-6.0 disodium hydrogen phosphate-
Potassium dihydrogen phosphate, acetic acid-sodium acetate, phosphoric acid-sodium phosphate, citric acid-sodium citrate and phosphoric acid hydrogen two
One or more in sodium-citric acid solution.
Method the most according to claim 3, it is characterised in that described buffer solution body
Quality/the Kg of long-pending/L and Herba Epimedii extract ratio is for 5-15:1.
Method the most according to claim 3, it is characterised in that the enzyme described in step a
The solution response time is 1-20 hour；Preferably, the described enzyme digestion reaction time is 10-20 hour；
Most preferably, the described enzyme digestion reaction time is 15-20 hour.
Method the most according to claim 1 and 2, it is characterised in that excessive in step a
The weight ratio of sheep leaves of pulse plants extract and beta-glycosidase is 1-50:1；It is highly preferred that excessive sheep in step a
The weight ratio of leaves of pulse plants extract and beta-glycosidase is 2-20:1；Most preferably, Herba Epimedii in step a
The weight ratio of extract and beta-glycosidase is 5-10:1.
Method the most according to claim 1, it is characterised in that in described step c,
First will filter after filtration cakes torrefaction, then pulverize, then will pulverize after filter cake be scattered in second
In alcohol, backflow is dissolved, and filters, and in filtrate, the volume/L of ethanol is equivalent to pulverize filter cake quality/Kg's
10-100 times.
Method the most according to claim 1, it is characterised in that in described step c,
By crystal crude product for the first time, add backflow in ethanol and dissolve, filtration, then filtrate cooling is tied again
Crystalline substance, the volume/L of refluxing ethanol is equivalent to 20-300 times of crystal crude product quality/Kg.
Method the most according to claim 1, it is characterised in that in described step a,
Described Herba Epimedii extract is prepared by ethonal extraction Herba Epimedii Chinese crude drug.
Method the most according to claim 9, it is characterised in that described alcohol solvent is
Ethanol water mixed solvent, the extract that ethanol water mixed solvent extraction Herba Epimedii Chinese crude drug obtains is first
Through concentrating, then pass through macroporous resin column absorption, then be the ethanol of 30%-100% by volumetric concentration
Aqueous solvent carries out eluting, obtains the Herba Epimedii extract for enzyme digestion reaction.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510129279.8A CN106148449B (en) | 2015-03-24 | 2015-03-24 | Preparation method of icariside I |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN114605483A (en) * | 2022-04-06 | 2022-06-10 | 贵州汇腾科技有限公司 | Preparation process of icariside I |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090170787A1 (en) * | 2005-11-30 | 2009-07-02 | Jun Seong Park | Cosmetic composition containing hydrolysates of icariin |
| CN101899077A (en) * | 2010-06-23 | 2010-12-01 | 吉林大学 | Extraction method and application of icariside I in Korean epimedium herb leaf |
| CN102311984A (en) * | 2011-07-05 | 2012-01-11 | 贾晓斌 | Method of preparing Baohuoside I from epimedium |
| CN102824394A (en) * | 2012-09-18 | 2012-12-19 | 西南民族大学 | Method for synchronously extracting and separating icariin and icarisid II from herba epimedii |
| CN103910772A (en) * | 2014-04-18 | 2014-07-09 | 成都合盛生物技术有限公司 | Method for simultaneously extracting icariin and baohuoside I and II from herba epimedii |
-
2015
- 2015-03-24 CN CN201510129279.8A patent/CN106148449B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090170787A1 (en) * | 2005-11-30 | 2009-07-02 | Jun Seong Park | Cosmetic composition containing hydrolysates of icariin |
| CN101899077A (en) * | 2010-06-23 | 2010-12-01 | 吉林大学 | Extraction method and application of icariside I in Korean epimedium herb leaf |
| CN102311984A (en) * | 2011-07-05 | 2012-01-11 | 贾晓斌 | Method of preparing Baohuoside I from epimedium |
| CN102824394A (en) * | 2012-09-18 | 2012-12-19 | 西南民族大学 | Method for synchronously extracting and separating icariin and icarisid II from herba epimedii |
| CN103910772A (en) * | 2014-04-18 | 2014-07-09 | 成都合盛生物技术有限公司 | Method for simultaneously extracting icariin and baohuoside I and II from herba epimedii |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN114605483A (en) * | 2022-04-06 | 2022-06-10 | 贵州汇腾科技有限公司 | Preparation process of icariside I |
| CN114605483B (en) * | 2022-04-06 | 2024-01-30 | 贵州汇腾科技有限公司 | Preparation process of icariside I |
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