CN105541938A - Method for preparing natural aucubin by membrane separation and macroporous adsorbent resin methods - Google Patents
Method for preparing natural aucubin by membrane separation and macroporous adsorbent resin methods Download PDFInfo
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- CN105541938A CN105541938A CN201510993202.5A CN201510993202A CN105541938A CN 105541938 A CN105541938 A CN 105541938A CN 201510993202 A CN201510993202 A CN 201510993202A CN 105541938 A CN105541938 A CN 105541938A
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- 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
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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
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Abstract
The invention relates to aucubin extracted from Eucommia ulmoides seeds and a preparing method thereof. The method comprises: using an organic solvent to extract glycoside components in the Eucommia ulmoides seeds, concentrating extract liquor under reduced pressure and then diluting, removing impurities from diluted liquor through a separating membrane, and separating and purifying through macroporous adsorbent resin to obtain an extract containing aucubin, drying the extract at high temperature under reduced pressure to obtain the aucubin product. The content is 95 percent or higher, and the method is efficient and simple and suitable for large scale industrial production.
Description
[technical field]
The present invention relates to a kind of effective ingredient in Chinese extractive technique field.More specifically, the present invention relates to a kind of aucubin of content more than 95%, also relate to the preparation method of described aucubin.
[background technology]
The bark of eucommia is a kind of famous and precious ancient Chinese medicinal materials, and its taste is sweet, warm in nature.Have tonify the liver and kidney, strengthen muscles and bones, recuperating CHONG and REN meridians, solid through antiabortive effect.Cortex Eucommiae extract also has the effects such as stronger hypertension, anti-help, anti-oxidant, anti-gastric-ulcer, and it can also promote collagen protein synthesis, also has other pharmacological effects many.The iridoids of the bark of eucommia and glucosides thereof are the main compound making it have these activity.The general bitter of iridoids material, but there is multiple physiologically active.Aucubin is a kind of iridoid glucosides separated from Folium Eucommiae, skin or seed.It has cholagogic and protects the liver, promotes the multiple pharmacological effect such as the synthesis of collagen protein, anti-muscle skeleton be aging.CN200910064817 discloses a kind of Cortex Eucommiae extract and is preparing the application had in the composition of sedative-hypnotic effect, also disclose Cortex Eucommiae extract for the preparation of the application in Cure for insomnia, anxiety or convulsions composition, this Cortex Eucommiae extract extracts continuously form through sherwood oil, ethyl acetate, propyl carbinol, second alcohol and water from Cortex Eucommiae, Folium Eucommiae, eucommia Bark male flower and/or bark of eucommia complete stool.CN201110202162 discloses Cortex Eucommiae extract for the preparation of skin anti-aging, the application especially in the makeup of anti-light aging or medicine.CN201110302835 relates to the bark of eucommia or Cortex Eucommiae extract for the preparation of the purposes in the medicine as vasoprotector and/or hypotensive agent, or for the preparation of the purposes treated and/or prevented in the medicine of vascular proliferative disease.CN201310027831 discloses a kind of Cortex Eucommiae extract and the application in the pharmaceutical composition preparing prevention and therapy bone metabolic disturbance disease thereof.
Membrane sepn is the separation technology occurred in 20 beginnings of the century.Membrane separation technique is owing to having separation, concentrated, purifying and refining function concurrently, there are again efficient, energy-saving and environmental protection, molecular level filters and filtration procedure is simple, be easy to the features such as control, therefore, food, medicine, biology, environmental protection, chemical industry, metallurgy, the energy, oil, water treatment, electronics, the field such as bionical are widely used in, create huge economic benefit and social benefit, become one of most important means in current separation science.
Macroporous adsorbent resin is the cross-linked polymer that a class does not contain ion-exchange group, and stable in physicochemical property, is insoluble to acid, alkali and organic solvent, has concentrated, centrifugation to organism, and by the interference of inorganic salts, strong ion and low molecular compound.Macroporous adsorbent resin can selectively adsorb organic compound matter by physical adsorption, thus can reach the object of separation and purification.According to resin surface character, macroporous adsorbent resin can be divided into nonpolar, Semi-polarity and polarity three class.
The separation purification method of high-purity aucubin is solvent extration, supercritical fluid extraction and column chromatography.With an organic solvent extraction process obtains the more of high-purity aucubin extract at present, but all need to use multiple inflammable and explosive organic solvent, require high to industrial equipments, operating process is complicated, the weak points such as product yield is not high, and waste liquid exhaust emission environment is serious.The weak point of supercritical fluid extraction is that plant and instrument cost is high, and energy consumption is serious.The main drawback of column chromatography is that silica gel can not regenerate, and production cost is high.
Therefore, also need a kind of both safety, product yield is high, environmental friendliness, production cost are low separation purification method at present, for this reason, the present inventor, on the basis of summing up prior art, completes the present invention finally through lot of experiments.
[summary of the invention]
[technical problem that will solve]
The object of this invention is to provide a kind of preparation method of aucubin.
[technical scheme]
The present invention is achieved through the following technical solutions.
The present invention relates to a kind of method extracting aucubin from seed of bark of eucommia.
The step of the method is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into seed of bark of eucommia powder, then according to weight ratio 1:5 ~ 10 of seed of bark of eucommia powder and alcohol or ketone aqueous solutions of organic solvent, at temperature 45 ~ 55 DEG C with the condition stirred, by concentration, 40 ~ 80% alcohol or ketone aqueous solutions of organic solvent extract 2.5 ~ 3.5 hours to described seed of bark of eucommia powder by volume, repeat extraction 2 ~ 3 times more in the same fashion, extracting solution merges, then concentrating under reduced pressure, obtains enriched material I, reclaims organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 2 ~ 4 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then described solution is allowed to be that the membrane module of the separatory membrane of 5000 ~ 50000 carries out membrane sepn by being equipped with molecular weight cut-off under the condition of pressure 0.1 ~ 0.5MPa, removing impurity, obtains a kind of filtrate;
C, resin absorption
The filtrate obtained according to allowing step B passes through macroporous adsorptive resins under the condition of flow velocity 0.5 ~ 2 times of column volume/h and room temperature, then use deionized water drip washing colourless to elutant, be 2 ~ 4 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 0.5 ~ 2 times of column volume/capable wash-out of h alcohol eluent with flow, collect elutriant, then concentrating under reduced pressure, obtains enriched material II, reclaims alcohol simultaneously;
D, oven dry
The enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry, obtains described aucubin.
According to a preferred embodiment of the invention a, in step, the granularity of described seed of bark of eucommia powder is 20 ~ 200 orders.
According to another preferred embodiment of the invention, in step, described organic solvent is a kind of organic solvent being selected from ethanol, methyl alcohol or acetone.
According to another preferred embodiment of the invention, in stepb, described separatory membrane is hollow-fibre membrane, ceramic membrane or polysulfone membrane.
According to another preferred embodiment of the invention, in step C, described macroporous adsorbent resin is other macroporous adsorbent resin that D101, HP20, XAD-2 macroporous adsorbent resin or its polarity are suitable with described macroporous adsorbent resin.
According to another preferred embodiment of the invention, in step C, described macroporous adsorptive resins is its height is the macroporous adsorptive resins of 15 ~ 25:1 with diameter ratio.
According to another preferred embodiment of the invention, in step C, described elutriant to be concentration be by volume 15 ~ 90% ethanol or methanol aqueous solution.
According to another preferred embodiment of the invention, in steps A and step C, described concentrating under reduced pressure concentrates under the condition of pressure 0.01 ~ 0.08MPa.
According to another preferred embodiment of the invention, in step D, it is dry at temperature 55 ~ 65 DEG C with the condition of pressure 0.01 ~ 0.05MPa that described high-temperature pressure-reduction is dried.
According to another preferred embodiment of the invention, in step D, the water-content of described aucubin is with aucubin total weight 3 ~ 5%.
In more detail the present invention will be described below.
The present invention adopts following technical scheme to prepare aucubin: get the dry bark of eucommia and to shell seed, pulverize, with organic solvent extraction 2 ~ 3 times, united extraction liquid concentrating under reduced pressure, the enriched material deionized water obtained dilutes, and then carries out membrane sepn, the filtrate obtained crosses macroporous adsorbent resin, first use polar organic solvent wash-out again with deionized water rinsing, elutriant carries out concentrating under reduced pressure, obtains peach jaurel glucoside extract.This extract is dried with high-temperature pressure-reduction again, obtains aucubin product.
Particularly, the present invention relates to a kind of method extracting aucubin from seed of bark of eucommia.The preparation process of this aucubin is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into seed of bark of eucommia powder, then according to weight ratio 1:5 ~ 10 of seed of bark of eucommia powder and alcohol or ketone aqueous solutions of organic solvent, at temperature 45 ~ 55 DEG C with the condition stirred, by concentration, 40 ~ 80% alcohol or ketone aqueous solutions of organic solvent extract 2.5 ~ 3.5 hours to described seed of bark of eucommia powder by volume, repeat extraction 2 ~ 3 times more in the same fashion, extracting solution merges, then concentrating under reduced pressure, obtains enriched material I, reclaims organic solvent simultaneously;
The bark of eucommia seed that shells is used in normally used disintegrating apparatus in Chinese medicinal materials manufacture field and pulverizes, and the granularity of described seed of bark of eucommia powder is generally 20 ~ 200 orders.Certainly, the granularity of seed of bark of eucommia powder is not very crucial, and the seed of bark of eucommia powder exceeding this size range also can use, but can affect extraction effect.
In the present invention, alcohol or ketone organic solvent should be appreciated that it is a kind of organic solvent that can extract aucubin from seed of bark of eucommia, and these organic solvents are a kind of volatile, and its residue also can not produce the organic solvent of adverse consequences to tobacco product.
In the present invention, described organic solvent is a kind of organic solvent being selected from ethanol, methyl alcohol or acetone.
Preferably, described organic solvent is ethanol or acetone.
More preferably, described organic solvent is ethanol.
In the present invention, the concentration of described organic solvent is by volume 40 ~ 80%, if this concentration is less than 40%, then can produce the water-soluble impurities such as a large amount of polysaccharide, protein, subsequent purification process brings difficulty; If this concentration is greater than 80%, then can the too much organic solvent of loss, increase production cost; Therefore, the concentration of described organic solvent is 40 ~ 80% is appropriate, preferably 48 ~ 70%, more preferably 56 ~ 62%.
The weight ratio of described organic solvent and alcohol or ketone aqueous solutions of organic solvent is 1:5 ~ 10, if exceed this weight ratio, then can dissolve too much impurity, and can increase solvent loss.
Described concentrating under reduced pressure concentrates under the condition of pressure 0.01 ~ 0.08MPa.The equipment used during concentrating under reduced pressure is normally used equipment in current processing technique of Chinese herbs field, such as rotatory evaporator, vacuum-concentrcted device.
Described organic solvent needs to reclaim after extraction step completes, and the distillation method usually adopting those skilled in the art to know reclaims.
B, membrane sepn
According to the volume ratio 2 ~ 4 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then described solution is allowed to be that the membrane module of the separatory membrane of 5000 ~ 50000 carries out membrane sepn by being equipped with molecular weight cut-off under the condition of pressure 0.1 ~ 0.5MPa, removing impurity, obtains a kind of filtrate;
When enriched material I deionized water dilutes, if the volume ratio of enriched material I and deionized water is less than 2, then can material viscosity excessive, separatory membrane pore volume easily blocks; If the volume ratio of enriched material I and deionized water is greater than 4, then can volume of material excessive, disengaging time is long; Therefore, the volume ratio of enriched material I and deionized water is 2 ~ 4 is rational.
In the present invention, compared with existing separation purification method, membrane separation technique high-level efficiency, less energy-consumption ground can remove in Chinese medical extract the small molecular weight impurities such as the macromole impurity component such as protein, polysaccharide that contains and inorganic salt.Therefore, membrane separation technique can significantly improve aucubin purity and quality for the present invention.
In the present invention, the separatory membrane of use is hollow-fibre membrane, ceramic membrane or polysulfone membrane.
Hollow-fibre membrane is a kind of outer likeness in form threadiness, has the film of self-supporting effect.Its tight zone can be positioned at fibrous outer surfaces, as reverse osmosis membrane, also can be positioned at the internal surface of fiber, as microfiltration membrane, and nanofiltration membrane and ultra-filtration membrane.The hollow-fibre membrane that the present invention uses is product sold in the market, the PVC hollow fiber membrane of the triumphant filter film technology company limited in such as Hangzhou.
Ceramic membrane is the solid film material belonged in membrane separation technique, mainly using inorganic ceramic materials such as the aluminum oxide of different size, zirconium white, titanium oxide and silicon oxide as supporter, forms through surface coatings, high-temperature firing.The ceramic membrane that the present invention uses is product sold, the such as ceramic membrane of Wuxi Kai Ou membrane separation plant company limited in the market.
Polysulfone membrane is the film having screening function that the macromolecular compound being repetition sulfuryl and arylidene by main chain is made.The sulphur atom of polysulfones is highest oxidation state, and the conjugative effect of sulfuryl makes it have excellent oxidation-resistance and thermostability, and ether chain improves the toughness of polysulfones, and phenyl ring improves mechanical strength and modulus.The polysulfone membrane that the present invention uses is product sold, the such as polysulphones hyperfiltration membrane of Hangzhou Hao Tianmo Science and Technology Ltd. in the market.
The present invention uses the molecular weight cut-off of separatory membrane to be 5000 ~ 50000, if separatory membrane molecular weight cut-off is too small, can increase disengaging time, cause the waste of time cost; Separatory membrane molecular weight cut-off is crossed conference and is introduced a large amount of macromole impurity, increases subsequent purification burden.To be the membrane module of the separatory membrane of 5000 ~ 50000 be product sold in the market, the PVC hollow fiber membrane of the triumphant filter film technology company limited in such as Hangzhou that the present invention uses and molecular weight cut-off is housed.
The present invention carries out membrane sepn under the condition of working pressure 0.1 ~ 0.5MPa; If described working pressure is lower than 0.1MPa, disengaging time is long; Higher than 0.5MPa, pressure of equipment is excessive, easily produces leakage.
C, resin absorption
The filtrate allowing step B obtain passes through macroporous adsorptive resins under the condition of flow velocity 0.5 ~ 2 times of column volume/h and room temperature, then use deionized water drip washing colourless to elutant, be 2 ~ 4 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 0.5 ~ 2 times of column volume/capable wash-out of h alcohol eluent with flow, collect elutriant, then concentrating under reduced pressure, obtains enriched material II, reclaims alcohol simultaneously;
In the present invention, macroporous adsorbent resin is that a class does not contain cation exchange groups and has the preparation of macroporous structure, has good macroreticular structure and larger specific surface area, can selectively by the organism in the physical adsorption aqueous solution.It has been widely used in the isolation andpurification process of Chinese herbal medicine effective ingredients.It is with vinylbenzene and acrylate for monomer, take divinylbenzene as linking agent, and with toluene, dimethylbenzene for pore-creating agent, their polymerizations that is cross-linked with each other define porous skeleton structure.This resin has adsorptivity by itself and by the Van der Waals force between the molecule that adsorbs and hydrogen bond action, there is because of its reticulated structure and high-specific surface area the ability of selectively adsorb organic compound matter, therefore use this resin can realize the object of isolation and purification.
In the present invention, the macroporous adsorbent resin used is D101, HP20 or XAD-2 type macroporous adsorbent resin, or other macroporous adsorbent resin that its polarity is suitable with described macroporous adsorbent resin, they are all extensive product solds in the market, the D101 type macroporous adsorbent resin such as produced by Tianjin Hao Ju resin Science and Technology Ltd., by the HP20 type macroporous adsorbent resin of Mitsubishi chemical production, the XAD-2 type macroporous adsorbent resin produced by Rhom and Hass of the U.S., the LX-11 produced by Xi'an Lanxiao Sci-Tech Co., Ltd., other macroporous adsorbent resin of D101C type.
The macroporous adsorptive resins that the present invention uses is its height is the macroporous adsorptive resins of 15 ~ 25:1 with diameter ratio.
The macroporous adsorbent resin that the present invention uses needs to carry out following pre-treatment before use:
95% alcohol immersion 24h, is washed till effluent liquid with 95% ethanol and water 1:5 is not muddy, washes with water to without alcohol taste; Pass through resin column with 5%HCl again, soak 2 ~ 4h, be washed to neutrality; Pass through resin column with 2%NaOH again, soak 2 ~ 4h, be washed to neutrality, for subsequent use.
Then, the filtrate allowing step B obtain by macroporous adsorptive resins, reaches and adsorbs completely under the condition of flow velocity 0.5 ~ 2 times of column volume/h and room temperature.
Described macroporous adsorptive resins first uses deionized water drip washing colourless to elutant after absorption, the water-soluble large polar impurity cleaning removing of the carbohydrate, mucus juice etc. that can more fully, up hill and dale macroporous adsorptive resins be adsorbed like this, then use alcohol eluent then.
In the present invention, described alcohol eluent is ethanol or methanol aqueous solution.The concentration of described ethanol or methanol aqueous solution is by weight by volume 15 ~ 90%.If when the concentration of described alcohol eluent is less than 15%, then eluent polarity is larger, target compound is difficult to wash-out, if when the concentration of described alcohol eluent is greater than 90%, then solvent loss is comparatively large, increases production cost, therefore, the concentration of described alcohol eluent is 15 ~ 90% is rational, preferably 20 ~ 65%, more preferably 35 ~ 50%.
Preferably, described alcohol eluent to be concentration be by weight 20 ~ 65% aqueous ethanolic solution.
During wash-out, eluent is slowly added in adsorption column, regulate flow velocity to reach 0.5 ~ 2 times of column volume/h.
It is identical that concentrating under reduced pressure situation and step B describe, and do not repeat them here.
D, oven dry
The enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry, obtains described aucubin.
It is dry at temperature 55 ~ 65 DEG C with the condition of pressure 0.01 ~ 0.05MPa that described high-temperature pressure-reduction is dried.
The Main Function of this baking step is to remove moisture in product, is convenient to storage.
In the present invention, it is product sold in the market that high-temperature pressure-reduction dries the equipment used, such as, by the vacuum drying oven of Shanghai precision instrumentation company limited.
Conventional H PLC method is adopted to analyze obtained aucubin product under the following conditions:
Use instrument: Japanese Shimadzu high performance liquid chromatograph EssentiaLC-15C
Measuring method: area external standard method
Condition determination: be analytical column with Nucleosi17-C18 (Φ 4.6 × 250mm, 10 μm), moving phase is methanol-water (5: 95); Column temperature: 25 DEG C; Determined wavelength: 203nm; Flow velocity: 1.0ml/min,
Measurement result shows, adopts the aucubin content of the inventive method products therefrom to be by weight more than 95%.
Extraction yield is calculated according to following equation by seed of bark of eucommia amount and aucubin product amount:
Extraction yield=(A × B)/(C × D) × 100%
A: quality product
B: aucubin content in product
C: raw materials quality
D: aucubin content in raw material
The extraction yield adopting the inventive method to extract seed of bark of eucommia is more than 95%.
[beneficial effect]
Prepared by the present invention high containing active constituent content in peach jaurel glucoside extract, and content is more than 95%.
Owing to selecting membrane separation technique, than existing separation purification method, it high-level efficiency, less energy-consumption ground can remove the small molecular weight impurity such as the macromole impurity component such as protein, polysaccharide and inorganic salt, can reach the object significantly improving aucubin purity and quality.
Owing to selecting macroporous adsorbent resin, it is good to the adsorption selectivity of aucubin, and absorption is fast, and desorb is also fast, and loading capacity is larger; Physical and chemical stability is high, and physical strength is good, and regeneration easily; Operate very easy, step is few, with low cost, extraction rate reached more than 95%, and solvent is recoverable all, is also applicable to large-scale commercial production.
[accompanying drawing explanation]
Fig. 1 is aucubin extractive HPLC figure prepared by the inventive method.
[embodiment]
The present invention will be illustrated in greater detail below by embodiment.
Embodiment 1: extract aucubin from seed of bark of eucommia
The implementation step of this embodiment is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into 80 order seed of bark of eucommia powder, then according to the weight ratio 1:9 of seed of bark of eucommia powder and alcohol or the ketone aqueous solution, described seed of bark of eucommia powder temperature 52 DEG C with stir condition under with concentration by volume 65% aqueous ethanolic solution extract 2.5 hours, repeat extraction 2 times more in the same fashion, extracting solution merges, then use the rotatory evaporator produced by Shanghai Yarong Biochemical Instrument Plant to carry out concentrating under reduced pressure under the condition of pressure 0.08MPa, obtain enriched material I, reclaim organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 2 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then described solution is allowed to be that the membrane module of the PVC hollow fiber membrane of being produced by the triumphant filter film technology company limited in Hangzhou of 5000 carries out membrane sepn by being equipped with molecular weight cut-off under the condition of pressure 0.1MPa, removing impurity, obtains a kind of filtrate;
C, resin absorption
The D101 type macroporous adsorptive resins of the filtrate allowing step B obtain by being produced by Tianjin Hao Ju resin Science and Technology Ltd. under the condition of flow velocity 1.5 times of column volume/h and room temperature, its height and diameter ratio are 22:1, then use deionized water drip washing colourless to elutant, be 3 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 1.4 times of column volumes/h concentration for 95% aqueous ethanolic solution wash-out agent row wash-out by volume with flow, collect elutriant, then the rotatory evaporator concentrating under reduced pressure under the condition of pressure 0.08MPa produced by Shanghai Yarong Biochemical Instrument Plant is used, obtain enriched material II, reclaim alcohol simultaneously,
D, oven dry
Use the vacuum drying oven produced by Shanghai precision instrumentation company limited, the enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry at temperature 55 DEG C with the condition of pressure 0.02MPa, and obtaining water-content is peach jaurel glucoside extract with aucubin total weight 3.0%.
Aucubin extractive HPLC analytical results prepared by this embodiment is see accompanying drawing 1.
The aucubin content adopting the method described in this specification sheets to measure its peach jaurel glucoside extract is 95.25%, and calculating extraction yield is 95.01%.
Embodiment 2: extract aucubin from seed of bark of eucommia
The implementation step of this embodiment is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into 140 order seed of bark of eucommia powder, then according to the weight ratio 1:10 of seed of bark of eucommia powder and alcohol or the ketone aqueous solution, described seed of bark of eucommia powder under the condition of temperature 45 C and stirring with concentration by volume 40% methanol aqueous solution extract 2.8 hours, repeat extraction 3 times more in the same fashion, extracting solution merges, then use the rotatory evaporator produced by Shanghai Yarong Biochemical Instrument Plant to carry out concentrating under reduced pressure under the condition of pressure 0.01MPa, obtain enriched material I, reclaim organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 3 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then allow described solution under the condition of pressure 0.4MPa by be equipped with molecular weight cut-off be 10000 carry out membrane sepn by the membrane module of the ceramic membrane of Wuxi Kai Ou membrane separation plant company limited, removing impurity, obtains a kind of filtrate;
C, resin absorption
The filtrate allowing step B obtain under the condition of flow velocity 1.8 times of column volume/h and room temperature by the HP20 type macroporous adsorptive resins by Mitsubishi chemical production, its height and diameter ratio are 25:1, then use deionized water drip washing colourless to elutant, be 2 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 0.5 times of column volume/h concentration for 1% ethanol or the capable wash-out of methanol aqueous solution eluent by volume with flow, collect elutriant, then the rotatory evaporator concentrating under reduced pressure under the condition of pressure 0.01MPa produced by Shanghai Yarong Biochemical Instrument Plant is used, obtain enriched material II, reclaim alcohol simultaneously,
D, oven dry
Use the vacuum drying oven produced by Shanghai precision instrumentation company limited, the enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry at temperature 58 DEG C with the condition of pressure 0.01MPa, and obtaining water-content is peach jaurel glucoside extract with aucubin total weight 3.6%.
Coming to the same thing of aucubin extractive HPLC analysis chart prepared by this embodiment and embodiment 1.
The aucubin content adopting the method described in this specification sheets to measure its peach jaurel glucoside extract is 95.43%, and calculating extraction yield is 95.06%.
Embodiment 3: extract aucubin from seed of bark of eucommia
The implementation step of this embodiment is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into 20 order seed of bark of eucommia powder, then according to the weight ratio 1:5 of seed of bark of eucommia powder and alcohol or the ketone aqueous solution, described seed of bark of eucommia powder temperature 48 DEG C with stir condition under with concentration by volume 45% aqueous acetone solution extract 3.2 hours, repeat extraction 2 times more in the same fashion, extracting solution merges, then use the rotatory evaporator produced by Shanghai Yarong Biochemical Instrument Plant to carry out concentrating under reduced pressure under the condition of pressure 0.02MPa, obtain enriched material I, reclaim organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 4 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then allow described solution under the condition of pressure 0.5MPa by be equipped with molecular weight cut-off be 30000 the membrane module of polysulphones hyperfiltration membrane produced by Hangzhou Hao Tianmo Science and Technology Ltd. carry out membrane sepn, removing impurity, obtains a kind of filtrate;
C, resin absorption
The XAD-2 type macroporous adsorptive resins of the filtrate allowing step B obtain by being produced by Rhom and Hass of the U.S. under the condition of flow velocity 2.0 times of column volume/h and room temperature, its height and diameter ratio are 15:1, then use deionized water drip washing colourless to elutant, be 3 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 0.8 times of column volume/h concentration for 35% aqueous ethanolic solution wash-out agent row wash-out by volume with flow, collect elutriant, then the rotatory evaporator concentrating under reduced pressure under the condition of pressure 0.02MPa produced by Shanghai Yarong Biochemical Instrument Plant is used, obtain enriched material II, reclaim alcohol simultaneously,
D, oven dry
Use the vacuum drying oven produced by Shanghai precision instrumentation company limited, the enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry at temperature 62 DEG C with the condition of pressure 0.04MPa, and obtaining water-content is peach jaurel glucoside extract with aucubin total weight 4.3%.
Coming to the same thing of aucubin extractive HPLC analysis chart prepared by this embodiment and embodiment 1.
The aucubin content adopting the method described in this specification sheets to measure its peach jaurel glucoside extract is 95.72%, and calculating extraction yield is 95.10%.
Embodiment 4: extract aucubin from seed of bark of eucommia
The implementation step of this embodiment is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into 200 order seed of bark of eucommia powder, then according to the weight ratio 1:7 of seed of bark of eucommia powder and alcohol or the ketone aqueous solution, described seed of bark of eucommia powder temperature 55 DEG C with stir condition under with concentration by volume 80% aqueous ethanolic solution extract 3.5 hours, repeat extraction 3 times more in the same fashion, extracting solution merges, then use the rotatory evaporator produced by Shanghai Yarong Biochemical Instrument Plant to carry out concentrating under reduced pressure under the condition of pressure 0.06MPa, obtain enriched material I, reclaim organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 3 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then described solution is allowed to be that the membrane module of the PVC hollow fiber membrane of being produced by the triumphant filter film technology company limited in Hangzhou of 50000 carries out membrane sepn by being equipped with molecular weight cut-off under the condition of pressure 0.2MPa, removing impurity, obtains a kind of filtrate;
C, resin absorption
The filtrate allowing step B obtain under the condition of flow velocity 1.4 times of column volume/h and room temperature by the HP20 type macroporous adsorptive resins by Mitsubishi chemical production, its height and diameter ratio are 18:1, then use deionized water drip washing colourless to elutant, be 4 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 2.0 times of column volumes/h concentration for 50% aqueous ethanolic solution wash-out agent row wash-out by volume with flow, collect elutriant, then the rotatory evaporator concentrating under reduced pressure under the condition of pressure 0.06MPa produced by Shanghai Yarong Biochemical Instrument Plant is used, obtain enriched material II, reclaim alcohol simultaneously,
D, oven dry
Use the vacuum drying oven produced by Shanghai precision instrumentation company limited, the enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry at temperature 65 DEG C with the condition of pressure 0.05MPa, and obtaining water-content is peach jaurel glucoside extract with aucubin total weight 5.0%.
Coming to the same thing of aucubin extractive HPLC analysis chart prepared by this embodiment and embodiment 1.
The aucubin content adopting the method described in this specification sheets to measure its peach jaurel glucoside extract is 95.89%, and calculating extraction yield is 95.14%.
Embodiment 5: extract aucubin from seed of bark of eucommia
The implementation step of this embodiment is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into 80 order seed of bark of eucommia powder, then according to the weight ratio 1:8 of seed of bark of eucommia powder and alcohol or the ketone aqueous solution, described seed of bark of eucommia powder temperature 48 DEG C with stir condition under with concentration by volume 80% aqueous acetone solution extract 2.5 hours, repeat extraction 3 times more in the same fashion, extracting solution merges, then use the rotatory evaporator produced by Shanghai Yarong Biochemical Instrument Plant to carry out concentrating under reduced pressure under the condition of pressure 0.02MPa, obtain enriched material I, reclaim organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 3 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then allow described solution under the condition of pressure 0.1MPa by be equipped with molecular weight cut-off be 10000 carry out membrane sepn by the membrane module of the polysulphones hyperfiltration membrane of Hangzhou Hao Tianmo Science and Technology Ltd., removing impurity, obtains a kind of filtrate;
C, resin absorption
The D101 type macroporous adsorptive resins of the filtrate allowing step B obtain by being sold by Tianjin Hao Ju resin Science and Technology Ltd. under the condition of flow velocity 1.0 times of column volume/h and room temperature, its height and diameter ratio are 23:1, then use deionized water drip washing colourless to elutant, be 3 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 1.4 times of column volumes/h concentration for 10% aqueous ethanolic solution wash-out agent row wash-out by volume with flow, collect elutriant, then the rotatory evaporator concentrating under reduced pressure under the condition of pressure 0.04MPa produced by Shanghai Yarong Biochemical Instrument Plant is used, obtain enriched material II, reclaim alcohol simultaneously,
D, oven dry
Use the vacuum drying oven produced by Shanghai precision instrumentation company limited, the enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry under the condition of temperature 60 C and pressure 0.03MPa, and obtaining water-content is peach jaurel glucoside extract with aucubin total weight 4.6%.
Coming to the same thing of aucubin extractive HPLC analysis chart prepared by this embodiment and embodiment 1.
The aucubin content adopting the method described in this specification sheets to measure its peach jaurel glucoside extract is 96.02%, and calculating extraction yield is 95.7%.
Embodiment 6: extract aucubin from seed of bark of eucommia
The implementation step of this embodiment is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into 140 order seed of bark of eucommia powder, then according to the weight ratio 1:6 of seed of bark of eucommia powder and alcohol or the ketone aqueous solution, described seed of bark of eucommia powder temperature 54 DEG C with stir condition under with concentration by volume 62% aqueous ethanolic solution extract 3.0 hours, repeat extraction 2 times more in the same fashion, extracting solution merges, then use the rotatory evaporator produced by Shanghai Yarong Biochemical Instrument Plant to carry out concentrating under reduced pressure under the condition of pressure 0.04MPa, obtain enriched material I, reclaim organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 2 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then described solution is allowed to be that the membrane module of the PVC hollow fiber membrane of being produced by the triumphant filter film technology company limited in Hangzhou of 30000 carries out membrane sepn by being equipped with molecular weight cut-off under the condition of pressure 0.4MPa, removing impurity, obtains a kind of filtrate;
C, resin absorption
The filtrate allowing step B obtain under the condition of flow velocity 1.2 times of column volume/h and room temperature by the HP20 type macroporous adsorptive resins by Mitsubishi chemical production, its height and diameter ratio are 20:1, then use deionized water drip washing colourless to elutant, be 4 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 1.5 times of column volumes/h concentration for 40% aqueous ethanolic solution wash-out agent row wash-out by volume with flow, collect elutriant, then the rotatory evaporator concentrating under reduced pressure under the condition of pressure 0.05MPa produced by Shanghai Yarong Biochemical Instrument Plant is used, obtain enriched material II, reclaim alcohol simultaneously,
D, oven dry
Use the vacuum drying oven produced by Shanghai precision instrumentation company limited, the enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry under the condition of temperature 60 C and pressure 0.03MPa, and obtaining water-content is peach jaurel glucoside extract with aucubin total weight 3.2%.
Coming to the same thing of aucubin extractive HPLC analysis chart prepared by this embodiment and embodiment 1.
The aucubin content adopting the method described in this specification sheets to measure its peach jaurel glucoside extract is 95.66%, and calculating extraction yield is 95.46%.
Claims (10)
1. extract a method for aucubin from seed of bark of eucommia, it is characterized in that the step of the method is as follows:
A, organic solvent extraction
Seed of bark of eucommia is shelled, is ground into seed of bark of eucommia powder, then according to weight ratio 1:5 ~ 10 of seed of bark of eucommia powder and alcohol or ketone aqueous solutions of organic solvent, at temperature 45 ~ 55 DEG C with the condition stirred, by concentration, 40 ~ 80% alcohol or ketone aqueous solutions of organic solvent extract 2.5 ~ 3.5 hours to described seed of bark of eucommia powder by volume, repeat extraction 2 ~ 3 times more in the same fashion, extracting solution merges, then concentrating under reduced pressure, obtains enriched material I, reclaims organic solvent simultaneously;
B, membrane sepn
According to the volume ratio 2 ~ 4 of enriched material I and deionized water, the enriched material I deionized water dilution that steps A obtains obtains a kind of solution, then described solution is allowed to be that the membrane module of the separatory membrane of 5000 ~ 50000 carries out membrane sepn by being equipped with molecular weight cut-off under the condition of pressure 0.1 ~ 0.5MPa, removing impurity, obtains a kind of filtrate;
C, resin absorption
The filtrate allowing step B obtain passes through macroporous adsorptive resins under the condition of flow velocity 0.5 ~ 2 times of column volume/h and room temperature, then use deionized water drip washing colourless to elutant, be 2 ~ 4 according to the volume ratio of alcohol eluent and macroporous adsorptive resins again, be 0.5 ~ 2 times of column volume/capable wash-out of h alcohol eluent with flow, collect elutriant, then concentrating under reduced pressure, obtains enriched material II, reclaims alcohol simultaneously;
D, oven dry
The enriched material II obtained by step C carries out high-temperature pressure-reduction oven dry, obtains described aucubin.
2. method according to claim 1, is characterized in that in step, and the granularity of described seed of bark of eucommia powder is 20 ~ 200 orders.
3. method according to claim 1, is characterized in that in step, and described organic solvent is a kind of organic solvent being selected from ethanol, methyl alcohol or acetone.
4. method according to claim 1, is characterized in that in stepb, and described separatory membrane is hollow-fibre membrane, ceramic membrane or polysulfone membrane.
5. method according to claim 1, is characterized in that in step C, and described macroporous adsorbent resin is other macroporous adsorbent resin that D101, HPD20, XAD-2 macroporous adsorbent resin or its polarity are suitable with described macroporous adsorbent resin.
6. method according to claim 1, is characterized in that in step C, and described macroporous adsorptive resins is its height is the macroporous adsorptive resins of 15 ~ 25:1 with diameter ratio.
7. extracting method according to claim 1, is characterized in that in step C, described elutriant to be concentration be by volume 15 ~ 90% ethanol or methanol aqueous solution.
8. extracting method according to claim 1, is characterized in that in steps A and step C, and described concentrating under reduced pressure concentrates under the condition of pressure 0.01 ~ 0.08MPa.
9. extracting method according to claim 1, is characterized in that in step D, and it is dry at temperature 55 ~ 65 DEG C with the condition of pressure 0.01 ~ 0.05MPa that described high-temperature pressure-reduction is dried.
10. extracting method according to claim 1, is characterized in that in step D, and the water-content of described aucubin is with aucubin total weight 3 ~ 5%.
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| CN101974049A (en) * | 2010-07-26 | 2011-02-16 | 南京泽朗农业发展有限公司 | Method for extracting aucubin from aucuba japonica leaves |
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| CN104546992A (en) * | 2015-01-30 | 2015-04-29 | 四川九章生物科技有限公司 | Folium cortex eucommiae extract as well as preparation method and application thereof |
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| CN101974049A (en) * | 2010-07-26 | 2011-02-16 | 南京泽朗农业发展有限公司 | Method for extracting aucubin from aucuba japonica leaves |
| CN102219814A (en) * | 2011-05-25 | 2011-10-19 | 河南大学 | Method for extracting aucubin from eucommia ulmoides oliver seed draff |
| CN103864867A (en) * | 2012-12-14 | 2014-06-18 | 湖北老龙洞杜仲开发有限公司 | Method for extracting aucubin from folium cortex eucommiae |
| CN104546992A (en) * | 2015-01-30 | 2015-04-29 | 四川九章生物科技有限公司 | Folium cortex eucommiae extract as well as preparation method and application thereof |
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