CN110343731A - A method of Herba Epimedii low sugar glycosides component is prepared from Herba Epimedii extraction - Google Patents
A method of Herba Epimedii low sugar glycosides component is prepared from Herba Epimedii extraction Download PDFInfo
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- CN110343731A CN110343731A CN201910674431.9A CN201910674431A CN110343731A CN 110343731 A CN110343731 A CN 110343731A CN 201910674431 A CN201910674431 A CN 201910674431A CN 110343731 A CN110343731 A CN 110343731A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/06—Benzopyran radicals
- C07H17/065—Benzo[b]pyrans
- C07H17/07—Benzo[b]pyran-4-ones
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
Abstract
The present invention, which relates to, discloses a kind of method for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, by external zymolysis technique, enzymatic hydrolysis desugar processing is carried out to the epimedium flavone polysaccharide glycosides component that alcohol extracting Herba Epimedii medicine materical crude slice obtains, obtains Herba Epimedii low sugar glycosides component to convert.The present invention is a kind of enzymatic hydrolysis high conversion rate, easy to operate, environmental-friendly, economically viable method, low sugar glycosides compound can be converted by the enzymatic hydrolysis completely of polysaccharide glycoside substance such as Epimedin A, Epimedin B, epimedin C and icariin etc. in barren wort total chromocor, precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B, rhamnopyranosyl icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I, with stable composed structure ratio, to improve the content of Herba Epimedii low sugar glycosides component.
Description
Technical field
The present invention relates to a kind of methods for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, belong to compound and extract system
Standby technical field.
Background technique
Elderly patients are easy to get osteoporosis very much, and China enters aging society, the patient of osteoporosis gradually
Quantity also can be more and more.Osteoporosis refers to that body bone is destroyed and can bring serious complication.Disease patient's bone is tough
Property reduce, easily occur systemic fracture, the age is about 55% in 60 years old or more crowd's disease incidence, and the incidence of fracture reaches
10%.Therefore, the drug for finding more efficient treatment bone loss disorders is then particularly important.
China's traditional Chinese medicine Herba Epimedii, has effects that kidney-replenishing, wind-damp dispelling, strengthening the bones and muscles, is usually used in treating osteoporosis
Disease, wherein the main active for playing drug effect is flavone compound.Flavone compound in Herba Epimedii medicine materical crude slice, mostly with sugar
The form of glycosides is present in medicine materical crude slice, different according to glycosyl number number is carried on parent nucleus, is divided into polysaccharide glycosides: Epimedin A, towards the leaves of pulse plants
Determine B, epimedin C and icariin etc. and low sugar glycosides: arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B, rhamnopyranosyl icariside I I and precious leaves of pulse plants glycosides I
Deng.With the increase for carrying glycosyl number, dissolubility increases, but absorbability is deteriorated, and the absorption of low sugar glycosides is better than the suction of polysaccharide glycosides
It receives, and action substance is Herba Epimedii low sugar glycosides.But polysaccharide glycosides chromocor compound content is relatively high in barren wort total chromocor, and medicine
It is less to imitate the low Glycosides Contents of better Herba Epimedii.Therefore, the method that demand improves Herba Epimedii low sugar glycosides yield, to further research and development
New drug with preferable function of resisting osteoporosis is of great significance.
Early-stage study shows oral barren wort total chromocor, by enterobacteriaceae metabolism facilitate epimedium brevicornum polysaccharide glycosides flavones to
The conversion of low sugar glycosides flavones, so that preferably absorbed into serum plays drug effect, but by the factors such as individual difference and intestinal flora be unstable,
Can not promote polysaccharide glycosides stable conversion is the better Herba Epimedii low sugar glycosides of drug effect.Secondly the side that research discovery is processed by high temperature
Method facilitates polysaccharide glycosides scission of link, desugar in barren wort total chromocor, converts to low sugar glycosides, but the complicated, higher cost by processing procedure,
And not can guarantee that polysaccharide glycosides is fully converted to Herba Epimedii low sugar glycosides, it is unfavorable for large-scale production.Traditional acid hydrolytic reaction, reaction
Excessively acutely, it is easy directly to slough the glycosyl connected on polysaccharide glycosides whole hydrolysis or even excessive hydrolysis destroys chemical component
Structure, the waste water that secondly complicated for operation and reaction generates are unfavorable for environmental protection.
Summary of the invention
Purpose: the present invention overcomes the lower deficiency of the low Glycosides Contents of Herba Epimedii in existing barren wort total chromocor, provides one kind
The method for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, by introducing external zymolysis technique, by hydrolase to excessive sheep
Polysaccharide glycosides in leaves of pulse plants general flavone carries out desugar enzymatic hydrolysis and is converted into Herba Epimedii low sugar glycosides, and has stable composed structure ratio, thus
Improve the yield with the active Herba Epimedii low sugar glycosides of more high-drug-effect.
The system of the better Herba Epimedii low sugar glycosides of drug effect is fully converted to using polysaccharide glycosides in catalysed in vitro barren wort total chromocor
Preparation Method is that have higher market development significance.And external zymolysis technique is introduced, it is taken off by commercial enzyme cheap and easy to get
Sugared enzyme digestion reaction, reaction condition is mild, easy to operate, and environmental pollution is small.And enzyme has high efficiency, safety and specificity strong
The characteristics of, high catalysis activity ensure that efficient catalytic effect can be realized in a small amount of enzyme preparation input amount；Enzyme itself
It is exactly protein, utilizations can be digested after high temperature or gastrointestinal tract denaturation, noresidue substance and potentially eats
Safety；It is compared with acid and alkali hydrolysis technique, enzyme has the specificity of specific recognition site, can only selectively slough specific site
Function group avoid excessively hydrolyzing, target chemical ingredient is destroyed to ensure that the integrality and accuracy of target product
The generation of phenomenon.
Technical solution: in order to solve the above technical problems, the technical solution adopted by the present invention are as follows:
A method of Herba Epimedii low sugar glycosides component is prepared from Herba Epimedii extraction characterized by comprising
(1) Herba Epimedii medicine materical crude slice is weighed, ethyl alcohol is added and carries out heating extraction, merging filtrate obtains barren wort total chromocor extraction
Liquid carries out that ethyl alcohol is recovered under reduced pressure, and concentrated extracting solution obtains epimedium flavone polysaccharide glycosides component and extracts concentrate；Wherein Herba Epimedii
Flavones polysaccharide glycosides component includes: Epimedin A, Epimedin B, epimedin C and icariin, and chemical structure characteristic is No. 3 positions
Contain multiple glucosides with No. 7 positions；Epimedin A, Epimedin B, epimedin C and icariin chemical structural formula:
(2) hydrolase cellulase, hydrolysis will be added in step (1) in gained epimedium flavone polysaccharide glycosides component concentrate
The ratio between enzyme quality and Herba Epimedii prepared slice quality are 0.2:20~2.0:20；Stirring adjusts pH4.5~7.0, and in 40~80 DEG C of temperature
Degree is lower to carry out 0.5~48h of enzyme digestion reaction, converts low sugar glycosides component for polysaccharide glycosides component, obtains enzyme digestion reaction liquid；
(3) enzyme digestion reaction liquid obtained by step (3) is subjected to centrifugal treating, abandons supernatant, obtains centrifugation；
(4) ethyl alcohol will be added in centrifugation obtained by step (4), stirs evenly, extracted, is centrifuged again, in collection
Clear liquid, is recovered under reduced pressure solvent to dry to get epimedium flavone low sugar glycosides component, including: precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, the arrow leaves of pulse plants
Glycosides B, rhamnopyranosyl icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I, chemistry
Structure feature is that disaccharide glycosides or monoglycosides are contained in No. 3 positions or No. 7 positions；Precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B, rhamnopyranosyl
The chemical structural formula of icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I:
Preferably, in epimedium flavone low sugar glycosides component obtained, precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B, mouse
Lee's glycosyl icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I content composed structure
It is respectively 5%~50%, 25%~95%, 5%~90%, 10%~80%, 1%~95% than range；5%~90%, institute
Stating percentage is mass percent；Precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B, rhamnopyranosyl icariside I I, dehydration Herba Epimedii
The sum of element -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I content are no more than 100%, and are greater than 80%.
Preferably, in step (1), the concentration of alcohol of addition is 40~80%, the material of ethyl alcohol and Herba Epimedii medicine materical crude slice
Liquor ratio is 15:1~25:1, is carried out heating extraction 1~3 time, and each extraction time is 0.5~3h.
Preferably, in step (1), the temperature that ethyl alcohol is recovered under reduced pressure is 40~80 DEG C, epimedium flavone polysaccharide glycosides
Component extracts concentrate to debita spissitudo, and every milliliter of prepared slice quality containing Herba Epimedii is 0.05~0.5g, i.e. Herba Epimedii medicine materical crude slice content
For 0.05~0.5g/mL.
Preferably, in step (2), speed of agitator is 20~200r/min.
Preferably, step (3), in step (4), the centrifugal rotational speed is 2000~10000r/min, from work
For preferred embodiment, in step (4), the concentration that ethyl alcohol is added is 50~100%, and the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality are
1~10mL/g.
The utility model has the advantages that a kind of method for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction provided by the invention, passes through
External zymolysis technique is introduced, promotes the polysaccharide glycosides in barren wort total chromocor that desugar enzymatic hydrolysis occurs by cellulase cheap and easy to get
Reaction, to the better Herba Epimedii low sugar glycosides conversion of drug activity, conversion ratio is higher, and has stable composed structure ratio, to mention
The high yield of Herba Epimedii low sugar glycosides, has ideal absorb using Herba Epimedii low sugar glycosides components composition obtained by the above method
Type and dissolubility, product purity fully demonstrate Chinese medicine multicomponent advantage 80% or more；Secondly W-response mild condition, behaviour
Make simplicity, it is environmental-friendly and low in cost, instrument and equipment is required lower, it is easy to accomplish large-scale industrial production.
Detailed description of the invention
Fig. 1 is the method flow diagram that embodiment prepares Herba Epimedii low sugar glycosides component from Herba Epimedii extraction；
Fig. 2 is high-efficient liquid phase chromatogram before and after the enzyme hydrolysis barren wort total chromocor that embodiment is prepared.
Specific embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and
It is apparent.But examples are merely exemplary for these, and it is not intended to limit the scope of the present invention in any way.Those skilled in the art
Member it should be understood that without departing from the spirit and scope of the invention can details to technical solution of the present invention and form into
Row modifications or substitutions, but these modifications and replacement are fallen within the protection scope of the present invention.
Fig. 1 is the method flow diagram that embodiment prepares Herba Epimedii low sugar glycosides component from Herba Epimedii extraction.
Embodiment 1:
(1) appropriate Herba Epimedii medicine materical crude slice is taken, 80% ethyl alcohol of 15 times of volumes is added, heating extraction 1 time, extracts 3h, filtering merges filter
Liquid carries out that ethyl alcohol is recovered under reduced pressure at 50 DEG C, and concentrated extracting solution to every milliliter of prepared slice quality containing Herba Epimedii is 0.1g, obtains excessive sheep
Leaves of pulse plants flavones polysaccharide glycosides component extracts concentrate；(2) ratio for being 0.2:20 according to the ratio between hydrolase quality and Herba Epimedii prepared slice quality
Example extracts concentrate to epimedium flavone polysaccharide glycosides component and cellulase is added, stir evenly, and adjusts pH value of solution to 4.5, if
Setting speed of agitator is 100r/min, and 40 DEG C of heating react 48h, obtain enzyme digestion reaction liquid, takes appropriate detection Epimedin A, determines towards the leaves of pulse plants
B, epimedin C and the substantially all disappearance of icariin, reach 99% enzymatic hydrolysis conversion ratio；(3) enzyme digestion reaction liquid is taken to be with revolving speed
10000r/min is centrifuged 10min, abandons supernatant, collects precipitating, is 10 according to the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality:
50% ethyl alcohol is added into centrifugation, stirs evenly for the ratio of 1mL/g, is again 10000r/min with revolving speed, centrifugation
10min collects supernatant, and ethyl alcohol is recovered under reduced pressure to dry to get Herba Epimedii low sugar glycosides component, (the 3.0g low sugar glycosides group of yield 3.0%
Point/100g medicine materical crude slice), polysaccharide glycosides converts the conversion ratio 95% of low sugar glycosides；Low Glycosides Contents 86.84% (precious leaves of pulse plants glycosides II 1.15%,
Arrow leaves of pulse plants glycosides A 3.82%, arrow leaves of pulse plants glycosides B 11.72%, rhamnopyranosyl icariside I I 19.00%, anhydroicartin -3-O-
Rhamnose-dideoxy furanose 3.02% and precious leaves of pulse plants glycosides I 47.53%).
Embodiment 2:
(1) appropriate Herba Epimedii medicine materical crude slice is taken, 40% ethyl alcohol of 25 times of volumes is added, heating extraction 1 time, extracts 2h, filtering merges filter
Liquid carries out that ethyl alcohol is recovered under reduced pressure at 60 DEG C, and concentrated extracting solution to every milliliter of prepared slice quality containing Herba Epimedii is 0.05g, is obtained excessive
Sheep leaves of pulse plants flavones polysaccharide glycosides component extracts concentrate；It (2) is 0.8:20's according to the ratio between hydrolase quality and Herba Epimedii prepared slice quality
Ratio extracts concentrate to epimedium flavone polysaccharide glycosides component and cellulase is added, stirs evenly, and adjusts pH value of solution to 6.5,
Setting speed of agitator is 50r/min, and 50 DEG C of heating reactions for 24 hours, obtain enzyme digestion reaction liquid, takes appropriate detection Epimedin A, determines towards the leaves of pulse plants
B, epimedin C and the substantially all disappearance of icariin, reach 100% enzymatic hydrolysis conversion ratio；(3) enzyme digestion reaction liquid is taken to be with revolving speed
8000r/min is centrifuged 20min, abandons supernatant, collects precipitating, is 8:1mL/ according to the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality
80% ethyl alcohol is added into centrifugation, stirs evenly for the ratio of g, is again 8000r/min with revolving speed, centrifugation 20min, receives
Collect supernatant, ethyl alcohol is recovered under reduced pressure to dry to get Herba Epimedii low sugar glycosides component, (the 2.5g low sugar glycosides component/100g of yield 2.5%
Medicine materical crude slice), polysaccharide glycosides converts the conversion ratio 96% of low sugar glycosides；Low Glycosides Contents 85.74% (precious leaves of pulse plants glycosides II 1.12%, arrow leaves of pulse plants glycosides A
3.78%, arrow leaves of pulse plants glycosides B 11.59%, rhamnopyranosyl icariside I I 18.78%, anhydroicartin -3-O- rhamnose -
Dideoxy furanose 3.01% and precious leaves of pulse plants glycosides I 47.46%).
Embodiment 3:
(1) appropriate Herba Epimedii medicine materical crude slice is taken, 50% ethyl alcohol of 20 times of volumes is added, heating extraction 2 times, extracts 1.5h, filtering merges
Filtrate carries out that ethyl alcohol is recovered under reduced pressure at 55 DEG C, and concentrated extracting solution to every milliliter of prepared slice quality containing Herba Epimedii is 0.2g, is obtained excessive
Sheep leaves of pulse plants flavones polysaccharide glycosides component extracts concentrate；It (2) is 0.4:20's according to the ratio between hydrolase quality and Herba Epimedii prepared slice quality
Ratio extracts concentrate to epimedium flavone polysaccharide glycosides component and cellulase is added, stirs evenly, and adjusts pH value of solution to 5.5,
Setting speed of agitator is 60r/min, and 40 DEG C of heating react 36h, obtain enzyme digestion reaction liquid, takes appropriate detection Epimedin A, determines towards the leaves of pulse plants
B, epimedin C and the substantially all disappearance of icariin, reach 99% enzymatic hydrolysis conversion ratio；(3) enzyme digestion reaction liquid is taken to be with revolving speed
3000r/min is centrifuged 40min, abandons supernatant, collects precipitating, is 5:1mL/ according to the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality
95% ethyl alcohol is added into centrifugation, stirs evenly for the ratio of g, is again 3000r/min with revolving speed, centrifugation 40min, receives
Collect supernatant, ethyl alcohol is recovered under reduced pressure to dry to get Herba Epimedii low sugar glycosides component, (the 2.6g low sugar glycosides component/100g of yield 2.6%
Medicine materical crude slice), polysaccharide glycosides converts the conversion ratio 97% of low sugar glycosides；Low Glycosides Contents 85.38% (precious leaves of pulse plants glycosides II 1.09%, arrow leaves of pulse plants glycosides A
3.74%, arrow leaves of pulse plants glycosides B 11.63%, rhamnopyranosyl icariside I I 18.76%, anhydroicartin -3-O- rhamnose -
Dideoxy furanose 2.99% and precious leaves of pulse plants glycosides I 47.17%).
Embodiment 4:
(1) appropriate Herba Epimedii medicine materical crude slice is taken, 70% ethyl alcohol of 20 times of volumes is added, heating extraction 3 times, extracts 0.5h, filtering merges
Filtrate carries out that ethyl alcohol is recovered under reduced pressure at 80 DEG C, and concentrated extracting solution to every milliliter of prepared slice quality containing Herba Epimedii is 0.5g, is obtained excessive
Sheep leaves of pulse plants flavones polysaccharide glycosides component extracts concentrate；It (2) is 1.0:20's according to the ratio between hydrolase quality and Herba Epimedii prepared slice quality
Ratio extracts concentrate to epimedium flavone polysaccharide glycosides component and cellulase is added, stirs evenly, and adjusts pH value of solution to 6.0,
Setting speed of agitator is 50r/min, and 75 DEG C of heating react 18h, obtain enzyme digestion reaction liquid, takes appropriate detection Epimedin A, determines towards the leaves of pulse plants
B, epimedin C and the substantially all disappearance of icariin, reach 98% enzymatic hydrolysis conversion ratio；(3) enzyme digestion reaction liquid is taken to be with revolving speed
2000r/min is centrifuged 40min, abandons supernatant, collects precipitating, is 2:1mL/ according to the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality
70% ethyl alcohol is added into centrifugation, stirs evenly for the ratio of g, is again 2000r/min with revolving speed, centrifugation 40min, receives
Collect supernatant, ethyl alcohol is recovered under reduced pressure to dry to get Herba Epimedii low sugar glycosides component, (the 2.9g low sugar glycosides component/100g of yield 2.9%
Medicine materical crude slice), polysaccharide glycosides converts the conversion ratio 97% of low sugar glycosides；Low Glycosides Contents 85.90% (precious leaves of pulse plants glycosides II 1.12%, arrow leaves of pulse plants glycosides A
3.79%, arrow leaves of pulse plants glycosides B 11.64%, rhamnopyranosyl icariside I I 19.01%, anhydroicartin -3-O- rhamnose -
Dideoxy furanose 2.97% and precious leaves of pulse plants glycosides I 47.37%).
Embodiment 5:
(1) appropriate Herba Epimedii medicine materical crude slice is taken, 50% ethyl alcohol of 22 times of volumes is added, heating extraction 2 times, extracts 2h, filtering merges filter
Liquid carries out that ethyl alcohol is recovered under reduced pressure at 40 DEG C, and concentrated extracting solution to every milliliter of prepared slice quality containing Herba Epimedii is 0.1g, obtains excessive sheep
Leaves of pulse plants flavones polysaccharide glycosides component extracts concentrate；(2) ratio for being 1.5:20 according to the ratio between hydrolase quality and Herba Epimedii prepared slice quality
Example extracts concentrate to epimedium flavone polysaccharide glycosides component and cellulase is added, stir evenly, and adjusts pH value of solution to 5.5, if
Setting speed of agitator is 50r/min, and 65 DEG C of heating react 3h, obtain enzyme digestion reaction liquid, take detect in right amount Epimedin A, Epimedin B,
Epimedin C and the substantially all disappearance of icariin, reach 98% enzymatic hydrolysis conversion ratio；(3) enzyme digestion reaction liquid is taken to be with revolving speed
5000r/min is centrifuged 30min, abandons supernatant, collects precipitating, is 4:1mL/ according to the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality
60% ethyl alcohol is added into centrifugation, stirs evenly for the ratio of g, is again 5000r/min with revolving speed, centrifugation 30min, receives
Collect supernatant, ethyl alcohol is recovered under reduced pressure to dry to get Herba Epimedii low sugar glycosides component, (the 2.8g low sugar glycosides component/100g of yield 2.8%
Medicine materical crude slice), polysaccharide glycosides converts the conversion ratio 96% of low sugar glycosides；Low Glycosides Contents 85.87% (precious leaves of pulse plants glycosides II 1.13%, arrow leaves of pulse plants glycosides A
3.77%, arrow leaves of pulse plants glycosides B 11.69%, rhamnopyranosyl icariside I I 18.95%, anhydroicartin -3-O- rhamnose -
Dideoxy furanose 2.88% and precious leaves of pulse plants glycosides I 47.45%).
Embodiment 6:
Appropriate Herba Epimedii medicine materical crude slice is taken, 55% ethyl alcohol of 18 times of volumes is added, heating extraction 2 times, extracts 2.5h, filtering merges filter
Liquid carries out that ethyl alcohol is recovered under reduced pressure at 60 DEG C, and concentrated extracting solution to every milliliter of prepared slice quality containing Herba Epimedii is 0.1g, obtains excessive sheep
Leaves of pulse plants flavones polysaccharide glycosides component extracts concentrate；(2) ratio for being 2.0:20 according to the ratio between hydrolase quality and Herba Epimedii prepared slice quality
Example extracts concentrate to epimedium flavone polysaccharide glycosides component and cellulase is added, stir evenly, and adjusts pH value of solution to 5.5, if
Setting speed of agitator is 50r/min, and 40 DEG C of heating react 12h, obtain enzyme digestion reaction liquid, take detect in right amount Epimedin A, Epimedin B,
Epimedin C and the substantially all disappearance of icariin, reach 100% enzymatic hydrolysis conversion ratio；(3) enzyme digestion reaction liquid is taken to be with revolving speed
6000r/min is centrifuged 20min, abandons supernatant, collects precipitating, is 6:1mL/ according to the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality
90% ethyl alcohol is added into centrifugation, stirs evenly for the ratio of g, is again 6000r/min with revolving speed, centrifugation 20min, receives
Collect supernatant, ethyl alcohol is recovered under reduced pressure to dry to get Herba Epimedii low sugar glycosides component, (the 2.9g low sugar glycosides component/100g of yield 2.9%
Medicine materical crude slice), polysaccharide glycosides converts the conversion ratio 96% of low sugar glycosides；Low Glycosides Contents 86.04% (precious leaves of pulse plants glycosides II 1.11%, arrow leaves of pulse plants glycosides A
3.79%, arrow leaves of pulse plants glycosides B 11.69%, rhamnopyranosyl icariside I I 18.95%, anhydroicartin -3-O- rhamnose -
Dideoxy furanose 3.01% and precious leaves of pulse plants glycosides I 47.49%).
Embodiment 7:
It is constituted using the ingredient of HPLC-ESI-QTOF-MS parsing Herba Epimedii low sugar glycosides component:
Chromatographic condition: chromatographic column is Agilent ZORBAX SB-C18 (250mm × 4.6mm, 5 μm)；Mobile phase be acetonitrile and
Water；Wavelength is 270nm；Column temperature is 30 DEG C；Flow velocity is 1mL/min；Sample volume is 10 μ L.Mobile phase is acetonitrile (A)-water, wherein 0
~45min, A:32.5%；45~46min, A:32.5-35%；46~61min, A:35%；61~62min, A:35-41%；
62~72min, A:41%；72~73min, A:41-32.5%.
Mass Spectrometry Conditions: electric spray ion source anionic textiles；Scanning mode: multiple reaction monitoring；Capillary voltage:
3500V；Mass range 80-1800m/z；Gas temperature: 320 DEG C；Fragmentation voltage: 100V；Dry gas stream amount: 8L/min；Atomization
Device pressure: 35psig；Collision voltage: 30V.Fig. 2 is high-efficient liquid phase chromatogram before and after embodiment enzyme hydrolysis barren wort total chromocor.
Mass spectrometric data is analyzed by MassHunter software as a result, acquiring the mass spectrum letter of the multiple ingredients of Herba Epimedii low sugar component
Breath is consulted about the chromocor compound pertinent literature having been found that in Herba Epimedii, the retention time of comprehensive analysis ingredient, opposite point
Protonatomic mass and fragment ion discrimination go out 6 main chemical compositions treasured leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, the arrow leaves of pulse plants in Herba Epimedii low sugar glycosides component
Glycosides B, rhamnopyranosyl icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I content groups
It than range is respectively 5mg/100mg low sugar glycosides component~50mg/100mg low sugar glycosides component at structure；25mg/100mg low sugar glycosides
Component~70mg/100mg low sugar glycosides component；5mg/100mg low sugar glycosides component~90mg/100mg low sugar glycosides component；10mg/
100mg low sugar glycosides component~80mg/100mg low sugar glycosides component；1mg/100mg low sugar glycosides component~95mg/100mg low sugar glycosides group
Point；5mg/100mg low sugar glycosides component~90mg/100mg low sugar glycosides component.
The discrimination of ingredient in 1 Herba Epimedii low sugar glycosides component of table
The above is only a preferred embodiment of the present invention, it should be pointed out that: for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (7)
1. a kind of method for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction characterized by comprising
(1) Herba Epimedii medicine materical crude slice is weighed, ethyl alcohol is added and carries out heating extraction, merging filtrate obtains barren wort total chromocor extracting solution, into
Ethyl alcohol is recovered under reduced pressure in row, and concentrated extracting solution obtains epimedium flavone polysaccharide glycosides component and extracts concentrate；Wherein epimedium flavone is more
Glucosides component includes: Epimedin A, Epimedin B, epimedin C and icariin；
(2) hydrolase cellulase, hydrolase matter will be added in gained epimedium flavone polysaccharide glycosides component concentrate in step (1)
The ratio between amount and Herba Epimedii prepared slice quality are 0.2: 20 ~ 2.0: 20；Stirring adjusts pH4.5 ~ 7.0, and in 40 ~ 80
0.5 ~ 48h of enzyme digestion reaction is carried out at a temperature of DEG C, is converted low sugar glycosides component for polysaccharide glycosides component, is obtained enzyme digestion reaction liquid；
(3) enzyme digestion reaction liquid obtained by step (3) is subjected to centrifugal treating, abandons supernatant, obtains centrifugation；
(4) ethyl alcohol will be added in centrifugation obtained by step (4), stirs evenly, extracted, is centrifuged again, collects supernatant,
Solvent is recovered under reduced pressure to dry to get epimedium flavone low sugar glycosides component, including: precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B,
Rhamnopyranosyl icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I.
2. the method according to claim 1 for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, which is characterized in that system
In the epimedium flavone low sugar glycosides component obtained, precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, arrow leaves of pulse plants glycosides B, rhamnopyranosyl icariside I I, dehydration
Icariine -3-O- rhamnose-dideoxy furanose and precious leaves of pulse plants glycosides I content composed structure are respectively 5% ~ 50% than range, 25% ~
95%, 5% ~ 90%, 10% ~ 80%, 1% ~ 95%；5% ~ 90%, the percentage is mass percent；Precious leaves of pulse plants glycosides II, arrow leaves of pulse plants glycosides A, the arrow leaves of pulse plants
Glycosides B, rhamnopyranosyl icariside I I, anhydroicartin -3-O- rhamnose-dideoxy furanose and treasured leaves of pulse plants glycosides I content it
Be no more than 100%, and be greater than 80%.
3. the method according to claim 1 for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, which is characterized in that step
Suddenly in (1), the concentration of alcohol of addition is 40 ~ 80%, and the solid-liquid ratio of ethyl alcohol and Herba Epimedii medicine materical crude slice is 15:1 ~ 25: 1, it carries out
Heating extraction 1 ~ 3 time, each extraction time is 0.5 ~ 3h.
4. the method according to claim 1 for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, which is characterized in that step
Suddenly in (1), the temperature that ethyl alcohol is recovered under reduced pressure is 40 ~ 80 DEG C, and epimedium flavone polysaccharide glycosides component extracts concentrate to appropriate dense
Degree, every milliliter of prepared slice quality containing Herba Epimedii are 0.05 ~ 0.5g, i.e., Herba Epimedii medicine materical crude slice content is 0.05 ~ 0.5g/mL.
5. the method according to claim 1 for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, which is characterized in that step
Suddenly in (2), speed of agitator is 20 ~ 200 r/min.
6. the method according to claim 1 for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, which is characterized in that step
Suddenly (3), in step (4), the centrifugal rotational speed is 2000 ~ 10000 r/min, and centrifugation time is 10 ~ 60 min.
7. the method according to claim 1 for preparing Herba Epimedii low sugar glycosides component from Herba Epimedii extraction, which is characterized in that step
Suddenly in (4), the concentration that ethyl alcohol is added is 50 ~ 100%, and the ratio between ethyl alcohol volume and Herba Epimedii prepared slice quality are 1 ~ 10 mL/g.
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