CN103709136A - Combined type method for extracting schisandrin b from fructus schisandrae - Google Patents
Combined type method for extracting schisandrin b from fructus schisandrae Download PDFInfo
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Abstract
The invention relates to a combined type method for extracting schisandrin b from fructus schisandrae. The method comprises the steps of pretreatment of raw materials, preparation of compound extractant, extraction and separation and purification; the compound extractant formed by appropriate ionic liquid, assistant and hexadecyl trimethyl ammonium bromide is adopted, solid sodium carbonate is added for auxiliary extraction, and besides, microwaves and expanded solvent are adopted for complementary extraction, therefore, the method for extracting the schisandrin b from the fructus schisandrae with high yield is achieved finally, the material utilization rage is improved, waste discharge and environment pollution, caused by a great amount of organic solvents, are avoided, and wide industrial application prospect is achieved.
Description
Technical field
The invention belongs to the technical field of extracting effective ingredient from natural phant.More specifically, the present invention relates to a kind of compounded methods of effectively extracting Wuweizisu B from Schisandra chinensis, there is good industrial application prospect.
Background technology
Shizandra berry is the mature fruit of magnoliaceae schisandra, it includes plurality of active ingredients, as lignanoid, triterpene, sesquiterpene, organic acid, volatilization wet goods composition, there is high nutritive value and pharmaceutical use, and show the multiple pharmacological effect to neural system, Digestive tract and cardiovascular systems.In recent years, Wuweizisu B receives much concern as a kind of important component in schisandra lignan material, content in Qi lignanoid is higher, biological activity is extensive, have anti-oxidant, remove free radical, reduce gpt, anticancer, the effect such as antibacterial, thereby become the study hotspot in the fields such as pharmacy, makeup.
In prior art, had the relevant report of multiple extraction Wuweizisu B, it mainly contains: solvent-extraction process, microwave extraction method, ultrasonic extraction, supercritical CO
2extraction method etc.For example:
A kind of method of extracting deoxyschizandrin and Wuweizisu B from shizandra berry has been reported in the patent application of CN102050707A, separated acquisition when it mainly adopts the solvent extraction method separated with macroporous resin adsorption to realize deoxyschizandrin with Wuweizisu B.
The Patent Application Publication of CN101070314A a kind of method of preparing high-purity deoxyschizandrin, it mainly adopts high-speed countercurrent chromatography, take normal hexane-methanol-water-acetonitrile as solvent systems, through flash liberation, can realize the separation of Wuweizisu B, and products obtained therefrom product degree is higher.
Japanese plum sound etc. (" research of ultrasonic extraction method Wuweizisu B technique ", the journal > > of < < Heilongjiang Bayi Agricultural Reclamation University, the 22nd the 2nd phase of volume, in April, 2010,74-79 page) adopt ultrasonic extraction method Wuweizisu B, and utilize quadratic regression equation quadrature rotation composite test to be optimized extraction process, show that optimum extraction condition is: ultrasonic 40min, solid-liquid ratio 1:10, alcohol concn is 90%, and Wuweizisu B extraction yield is 2.11mg/g.
Meng Xianjun etc. (" research of supercritical fluid extraction Wuweizisu B ", < < food research and development > >, the 30th the 3rd phase of volume, in March, 2009,37-39 page) employing supercritical CO
2extraction Wuweizisu B, and adopt the method for orthogonal experiment to inquire into pressure, temperature, time, the impact of granularity on Wuweizisu B extraction yield, determine that optimum process condition is: pressure 30MPa, 40 ℃ of temperature, granularity 40 orders, time 2h.
Yet, although scientific research personnel has been developed the technique of numerous extraction Wuweizisu Bs, but all more or less there are problems in existing extraction or separating technology, large such as solvent load, extraction time is long, extraction cost is high, extraction yield is lower etc., visible these techniques are still urgently improved, to meet the ultimate aim of suitability for industrialized production and application, realize the high yield acquisition of maximum using and the product of material.Therefore, develop the pressing problem that a kind of technique of extracting efficiently Wuweizisu B becomes industrial circle.
For the defect existing in prior art, the inventor is through a large amount of creative researches and experiment condition screening, be intended to develop a kind of hybrid process that extracts Wuweizisu B from Schisandra chinensis, fully meet the demand of large-scale commercial production, realize that cost is low, yield is high and the object of environmental protection.
Summary of the invention
The many defects that exist for prior art, the inventor, through a large amount of further investigations, after having paid sufficient creative work, has developed a kind of highly effective extraction method of Wuweizisu B.
Particularly, the invention provides a kind of compounded methods of extracting Wuweizisu B from Schisandra chinensis, described method comprises the steps:
A, raw materials pretreatment;
The preparation of B, compound extractant;
C, extraction operation;
D, separation and purification.
More specifically, the invention provides a kind of compounded methods of extracting Wuweizisu B from Schisandra chinensis, described method comprises the steps:
A, raw materials pretreatment
Get dry Schisandra chinensis, with 50-60 ℃ of warm water washing, then filter, dry, pulverized again 50-80 mesh sieve, then to after adding sodium carbonate solid in the gained powder that sieves and being fully uniformly mixed, then with vibration mill machinery pulverizing 5-10min, obtain Chinese Magnolivine Fruit.
The preparation of B, compound extractant
Ionic liquid and auxiliary agent are proportionally uniformly mixed, then add cetyl trimethylammonium bromide, under room temperature, continue to stir 15-25min, mix, obtain required compound extractant.
C, extraction operation
Take the Chinese Magnolivine Fruit of steps A, the compound extractant that adds wherein step B to make, stirs 20-30min, is then placed in microwave extraction still and extracts, and last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to expanded solvents system, is forced into 5-6MPa and forms expanded solvents system, stirring and leaching 30-40min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.
D, separation and purification
The extracting solution of step C gained being carried out to vacuum concentration, add petroleum ether dissolution, adopt silica gel column chromatography, is the petroleum ether-ethyl acetate wash-out of 3:1 (being that both volume ratio v/v are 3:1) by volume ratio, collects elutriant, and evaporation obtains crude product except desolventizing;
Crude product is carried out to separation with high speed adverse current chromatogram, solvent system is normal hexane-ethyl acetate-methanol-water, first the volume ratio with 16-20:1:20:1.2 is disposed at above-mentioned solvent system in separating funnel, shake up static layering, place 12-18h, getting upper strata is stationary phase, lower floor is moving phase, make sampling valve for sample introduction state, stationary phase is filled to chromatography column with pump, opening speed controller, make the chromatographic column forward of high-speed counter-current chromatograph, turn up 800r/min, get stationary phase and dissolve crude product to 20mg/ml, sample injection, flow rate of mobile phase is 2ml/min, according to detector ultraviolet spectrogram, receive Wuweizisu B component, vacuum concentration obtains Wuweizisu B sterling, and use HPLC to carry out purity check.
In the described compounded methods of extraction Wuweizisu B according to the present invention, the addition of sodium carbonate described in described steps A, for the 1-5% of dry Schisandra chinensis weight, is preferably 3%.The use of sodium carbonate can effectively reduce the foreign matter content such as organic acid, tannin in product, can accelerate to destroy solid material surface simultaneously, makes to be easier to disperse between particle, plays the auxiliary effect of pulverizing.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, ionic liquid in described step B is 1-butyl-2,3-methylimidazole chlorate, 1-butyl-2,3-methylimidazole Bromide, 1-butyl-2, any one in 3-methylimidazole tosilate.The inventor creatively adopts tri-substituted imidazole class ionic liquid to be applied among extraction agent, and the harm of effectively having avoided the use of a large amount of organic solvents to cause environment has obtained the technique effect of excellent yield simultaneously unexpectedly.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, the auxiliary agent in described step B is any one in diethylene glycol monomethyl ether, diethylene glycol dimethyl ether, dipropylene glycol monomethyl ether, dipropylene glycol dme.In the present invention, the selection of auxiliary agent relates to mating of target molecule and the intermolecular hole of double solvents first, second played the dispersive ability that reduces viscosity of il and contribute to improve material and solvent, the inventor just filters out applicable compound auxiliary through great many of experiments, and in this system, not ether or polyethers all can reach above-mentioned technique effect arbitrarily.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, the weight ratio of described step B intermediate ion liquid and auxiliary agent is 1:1-5, for example, can be 1:1,1:2,1:3,1:4 or 1:5, is preferably 1:3.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, the 3-5% that in described step B, the weight of cetyl trimethylammonium bromide is ionic liquid, for example, can be 3%, 3.5%, 4%, 4.5% or 5%, is preferably 4%.Cetyl trimethylammonium bromide can dissolved cell film, discharge bioactive molecule, and in this system, helps lend some impetus to the separated of target molecule or shift.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, in described step C, the weight ratio of Chinese Magnolivine Fruit and compound extractant is 1:6-10, can be to indefiniteness 1:6,1:7,1:8,1:9 or 1:10, is preferably 1:8.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, in described step C, microwave extraction parameter is: microwave 350-550W, time 5-15min; Preferred microwave 400W, time 10min.
In the described compounded methods of extraction Wuweizisu B according to the present invention, as a kind of preferred implementation, in described step C, the volume of expanded solvents consists of: 30% ethanol and 70%CO
2, the usage ratio of expanded solvents and Chinese Magnolivine Fruit is 2-4ml/g (its implication is the expanded solvents that every g Chinese Magnolivine Fruit is used 2-4ml), is preferably 3ml/g.
In the described compounded methods of extraction Wuweizisu B according to the present invention, in described step D, the analysis condition of HPLC is: Water C18 chromatographic column (4.6mm * 250mm, 5 μ m), column temperature: 25 ℃, moving phase: methyl alcohol/0.5% aqueous acetic acid (being that mass concentration is 0.5% aqueous acetic acid)=3/1 (v/v), flow velocity: 1.0ml/min, sample size: 10 μ L, detect wavelength: 220nm.
Except the ultraviolet spectrogram with standard Wuweizisu B contrasts, product separation of the present invention further identifies by mass spectrum, and result shows that the object product obtaining is Wuweizisu B.
As mentioned above, the invention provides a kind of novel method of extracting Wuweizisu B from Schisandra chinensis, compared with prior art, the present invention has produced following beneficial effect:
(1) inventor creatively prepares and is applicable to the compound extractant that Wuweizisu B extracts, and it comprises a certain proportion of ionic liquid, auxiliary agent and cetyl trimethylammonium bromide, and this extraction agent shows excellent extraction effect;
(2) take the lead in adopting the second extraction pattern of microwave/extraction agent+expanded solvents lixiviate, greatly improved the utilization ratio of material and obtained more quality products;
(3) add solid sodium carbonate before pulverizing, accelerating solid surface breakdown and auxiliary pulverizing can effectively be reduced the content of the impurity such as organic acid, tannin simultaneously in leaching process.
(4) by using high speed adverse current chromatogram, separation has obtained Wuweizisu B.
By the Combinatorial Optimization of technique and the selection of parameter, and supporting the use of extraction agent and obtained excellent extraction effect, thereby for meeting large-scale industrial production, lay a solid foundation, there is market using value widely.
Embodiment
Below by specific embodiment, the present invention is described in detail; but the purposes of these exemplary embodiments and object are only used for exemplifying the present invention; not real protection scope of the present invention is formed to any type of any restriction, more non-protection scope of the present invention is confined to this.
Embodiment 1: the kind of additive and consumption in steps A
A, raw materials pretreatment
Get the Schisandra chinensis that 1kg is dry, with 55 ℃ of warm water washings, then filter, dry, pulverized again 50 mesh sieves, then to after adding in the gained powder that sieves and being fully uniformly mixed after the additive of different sorts and consumption, then pulverize 5-10min with vibration mill machinery, obtain Chinese Magnolivine Fruit.Wherein, the kind of additive and consumption see the following form 1.
The preparation of B, compound extractant
By ionic liquid 1-butyl-2,3-methylimidazole chlorate and auxiliary agent diethylene glycol dimethyl ether are uniformly mixed according to weight ratio 1:3, then add the cetyl trimethylammonium bromide that is equivalent to ionic liquid weight 4%, under room temperature, continue to stir 20min, mix, obtain required compound extractant.
C, extraction operation
Take the Chinese Magnolivine Fruit of 100g steps A, the compound extractant that adds wherein 800g step B to make, stirs 25min, and be then placed in microwave extraction still and extract, microwave 400W, time 10min, last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to 300ml expanded solvents system, is forced into 5.5MPa and forms expanded solvents system, stirring and leaching 35min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.Wherein, described swollen solvent volume consists of: 30% ethanol and 70%CO
2.
D, separation and purification
The extracting solution of step C gained is carried out to vacuum concentration, add petroleum ether dissolution, adopt silica gel column chromatography, the petroleum ether-ethyl acetate wash-out that is 3:1 by volume ratio, collects elutriant, and evaporation obtains crude product except desolventizing;
Crude product is carried out to separation with high speed adverse current chromatogram, solvent system is normal hexane-ethyl acetate-methanol-water, first the volume ratio with 8:1:20:1.2 is disposed at above-mentioned solvent system in separating funnel, shake up static layering, place 15h, getting upper strata is stationary phase, lower floor is moving phase, make sampling valve for sample introduction state, stationary phase is filled to chromatography column with pump, opening speed controller, make the chromatographic column forward of high-speed counter-current chromatograph, turn up 800r/min, get stationary phase and dissolve crude product to 20mg/ml, sample injection, flow rate of mobile phase is 2ml/min, according to detector ultraviolet spectrogram, receive Wuweizisu B component, vacuum concentration obtains Wuweizisu B sterling, and use HPLC to carry out purity check.
The HPLC analysis condition of described sterling is: Water C18 chromatographic column (4.6mm * 250mm, 5 μ m), column temperature: 25 ℃, moving phase: methyl alcohol/0.5% aqueous acetic acid (being that mass concentration is 0.5% aqueous acetic acid)=3/1 (v/v), flow velocity: 1.0ml/min, sample size: 10 μ L, detect wavelength: 220nm.
The analytical data of mass spectrum of gained sterling is as follows: MS m/z:400.03 (M+1,100).
In steps A, use different additives and during with respect to the different amounts of dry Schisandra chinensis, the result of its final products therefrom total amount and purity is as shown in table 1 below:
Table 1. additive types and consumption
"-" represents not add additive
As seen from the above table, by adding additive, can improve the ultimate capacity of products therefrom, the use of wherein finding sodium carbonate can effectively reduce organic acid in product, the foreign matter contents such as tannin, can accelerate to destroy solid material surface simultaneously, make to be easier to disperse between particle, play the auxiliary effect of pulverizing, thereby improved the output of Wuweizisu B, and other inorganic salt (as calcium hydroxide) are added in selection, sugar cube (as sucrose) or even while belonging to the salt of wormwood of a class together with sodium carbonate, all can not reach identical extraction effect, the use of visible cheap sodium carbonate is of great advantage, there is best technique effect.
Embodiment 2: the kind of extracting ionic liquid in step B
A, raw materials pretreatment
Get the Schisandra chinensis that 1kg is dry, with 60 ℃ of warm water washings, then filter, dry, pulverized again 60 mesh sieves, then to after being added in the gained powder that sieves and being fully uniformly mixed after the sodium carbonate solid of dry Schisandra chinensis weight 3%, with vibration mill machinery, pulverize 5-10min again, obtain Chinese Magnolivine Fruit.
The preparation of B, compound extractant
Different types of ionic liquid and auxiliary agent diethylene glycol dimethyl ether are uniformly mixed according to weight ratio 1:3, then add the cetyl trimethylammonium bromide that is equivalent to ionic liquid weight 4%, under room temperature, continue to stir 25min, mix, obtain required compound extractant.Wherein, the kind of ionic liquid sees the following form 2.
C, extraction operation
Take the Chinese Magnolivine Fruit of 100g steps A, the compound extractant that adds wherein 800g step B to make, stirs 25min, and be then placed in microwave extraction still and extract, microwave 400W, time 10min, last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to 300ml expanded solvents system, is forced into 5MPa and forms expanded solvents system, stirring and leaching 40min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.Wherein, described expanded solvents volume consists of: 30% ethanol and 70%CO
2.
D, separation and purification
Adopt the identical separation step in the embodiment of the present invention 1 and obtain Wuweizisu B sterling.
While using different ionic liquids in step B, the result of its final products therefrom total amount and purity is as shown in table 2 below:
The kind of table 2. ionic liquid
As seen from the above table, the suitable extraction agent main ingredient as Wuweizisu B of tri-substituted imidazole class ionic liquid, especially 1-butyl-2,3-methylimidazole chlorate has best extraction effect (will higher than corresponding bromine salt), it shows more excellent effect with respect to triethyl ammonium chlorate and disubstituted imidazole class chlorate, and ionic liquid replaces the use of a large amount of organic solvents to have obvious environmental-protecting performance.
Embodiment 3: the kind of auxiliary agent
A, raw materials pretreatment
Get the Schisandra chinensis that 1kg is dry, with 50 ℃ of temperature launderings, then filter, dry, pulverizing 70 mesh sieves, then to after being added in the gained powder that sieves and being fully uniformly mixed after the sodium carbonate solid of dry Schisandra chinensis weight 3%, with vibration mill machinery, pulverize 5-10min again, obtain Chinese Magnolivine Fruit.
The preparation of B, compound extractant
By ionic liquid 1-butyl-2,3-methylimidazole chlorate and auxiliary agent are uniformly mixed according to weight ratio 1:3, then add the cetyl trimethylammonium bromide that is equivalent to ionic liquid weight 4%, continue to stir 20min under room temperature, mix, obtain required compound extractant.Wherein, the kind of auxiliary agent sees the following form 3.
C, extraction operation
Take the Chinese Magnolivine Fruit of 100g steps A, the compound extractant that adds wherein 800g step B to make, stirs 25min, and be then placed in microwave extraction still and extract, microwave 400W, time 10min, last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to 350ml expanded solvents system, is forced into 6MPa and forms expanded solvents system, stirring and leaching 40min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.Wherein, described expanded solvents volume consists of: 30% ethanol and 70%CO
2.
D, separation and purification
Adopt the identical separation step in the embodiment of the present invention 1 and obtain Wuweizisu B sterling.
While using different auxiliary agent in step B, the result of its final products therefrom total amount and purity is as shown in table 3 below:
The kind of table 2. auxiliary agent
As seen from the above table, traditional cyclic ethers, polyethers are not all the optimal selections of extracting auxiliary agent, the ethers of contrary short chain has but played the effect that remarkable promotion is extracted, this may be not suitable for extracting Wuweizisu B in the solvent hole based on cyclic ethers and ionic liquid formation, and polyethers causes solution to exist certain viscosity to be unfavorable for the diffusion of the solvent molecule of material because segment is long.Wherein diethylene glycol dimethyl ether has best extraction effect.
Embodiment 4: the consumption of auxiliary agent
A, raw materials pretreatment
Get the Schisandra chinensis that 1kg is dry, with 60 ℃ of warm water washings, then filter, dry, pulverized again 80 mesh sieves, then to after being added in the gained powder that sieves and being fully uniformly mixed after the sodium carbonate solid of dry Schisandra chinensis weight 3%, with vibration mill machinery, pulverize 5-10min again, obtain Chinese Magnolivine Fruit.
The preparation of B, compound extractant
By ionic liquid 1-butyl-2, the auxiliary agent diethylene glycol dimethyl ether of 3-methylimidazole chlorate and different amounts is uniformly mixed, and then adds the cetyl trimethylammonium bromide that is equivalent to ionic liquid weight 4%, continues to stir 20min under room temperature, mix, obtain required compound extractant.
C, extraction operation
Take the Chinese Magnolivine Fruit of 100g steps A, the compound extractant that adds wherein 800g step B to make, stirs 25min, and be then placed in microwave extraction still and extract, microwave 450W, time 15min, last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to 300ml expanded solvents system, is forced into 5.5MPa and forms expanded solvents system, stirring and leaching 40min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.Wherein, described expanded solvents volume consists of: 30% ethanol and 70%CO
2.
D, separation and purification
Adopt the identical separation step in the embodiment of the present invention 1 and obtain Wuweizisu B sterling.
While using the extraction auxiliary agent of different amounts in step B, the result of its final products therefrom total amount and purity is as shown in table 4 below:
The consumption of table 4. auxiliary agent
As seen from the above table, the ratio of ionic liquid and auxiliary agent is one of key factor regulating compound extractant overall performance, amount of auxiliary increase can effectively promote the dispersion of material, make target molecule be easy to same solvent molecule effect and be transferred in solvent phase, and the amount of the auxiliary agent difference of extraction agent system to target molecule solubility property that all can cause less than normal or bigger than normal has best extraction effect when wherein ionic liquid and auxiliary agent weight ratio are 1:3.
Embodiment 5: microwave extraction condition
A, raw materials pretreatment
Get the Schisandra chinensis that 1kg is dry, with 55 ℃ of warm water washings, then filter, dry, pulverized again 60 mesh sieves, then to after being added in the gained powder that sieves and being fully uniformly mixed after the sodium carbonate solid of dry Schisandra chinensis weight 3%, with vibration mill machinery, pulverize 5-10min again, obtain Chinese Magnolivine Fruit.
The preparation of B, compound extractant
By ionic liquid 1-butyl-2,3-methylimidazole chlorate and auxiliary agent diethylene glycol dimethyl ether are uniformly mixed according to weight ratio 1:3, then add the cetyl trimethylammonium bromide that is equivalent to ionic liquid weight 4%, under room temperature, continue to stir 20min, mix, obtain required compound extractant.
C, extraction operation
Take the Chinese Magnolivine Fruit of 100g steps A, the compound extractant that adds wherein 800g step B to make, stirs 25min, is then placed in microwave extraction still and extracts, and last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to 300ml expanded solvents system, is forced into 5MPa and forms expanded solvents system, stirring and leaching 30min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.Wherein, described expanded solvents volume consists of: 30% ethanol and 70%CO
2, microwave extraction condition is as shown in table 5 below.
D, separation and purification
Adopt the identical separation step in the embodiment of the present invention 1 and obtain Wuweizisu B sterling.
While using different microwave powers and time in step C, the result of its final products therefrom total amount and purity is as shown in table 5 below:
Table 5. microwave power and time
As seen from the above table, microwave power has comparatively significantly impact to extraction effect, extend extraction time to extract yield without remarkable effect.When microwave power is, while 400W and time being 10min, to have best extraction effect.
In sum, the inventor is by a large amount of theoretical investigationes and experimental exploring, creationary screening has also been prepared the compound extractant that is suitable for extracting Wuweizisu B, it consists of specific ionic liquid, auxiliary agent and cetyl trimethylammonium bromide, and adopt the further assisted extraction of sodium carbonate additive, in addition use the complementary extraction formula of microwave and expanded solvents means, thereby finally realized the extraction Wuweizisu B of high yield from Schisandra chinensis, improved the utilization ratio of material, the environmental pollution of simultaneously having avoided a large amount of organic solvents to cause.Wherein, any processing parameter is selected and the combination of the means of extraction is all a large amount of creative work gained of the inventor, and this extraction process has obtained the technique effect of high extraction also beyond expectationly, has industrial applications prospect and marketable value widely.
The purposes that should be appreciated that these embodiment only limits the scope of the invention for the present invention being described but not being intended to.In addition; also should understand; after having read technology contents of the present invention, those skilled in the art can make various changes, modification and/or modification to the present invention, within these all equivalent form of values fall within the protection domain that the application's appended claims limits equally.
Claims (10)
1. from Schisandra chinensis, extract a compounded methods for Wuweizisu B, described method comprises the steps:
A, raw materials pretreatment;
The preparation of B, compound extractant;
C, extraction operation;
D, separation and purification.
2. method according to claim 1, is characterized in that: described method comprises the steps:
A, raw materials pretreatment
Get dry Schisandra chinensis, with 50-60 ℃ of warm water washing, then filter, dry, pulverized again 50-80 mesh sieve, then to after adding sodium carbonate solid in the gained powder that sieves and being fully uniformly mixed, then with vibration mill machinery pulverizing 5-10min, obtain Chinese Magnolivine Fruit.
The preparation of B, compound extractant
Ionic liquid and auxiliary agent are proportionally uniformly mixed, then add cetyl trimethylammonium bromide, under room temperature, continue to stir 15-25min, mix, obtain required compound extractant.
C, extraction operation
Take the Chinese Magnolivine Fruit of steps A, the compound extractant that adds wherein step B to make, stirs 20-30min, is then placed in microwave extraction still and extracts, and last filtering separation obtains filter residue and filtrate 1; Filter residue is placed in to expanded solvents system, is forced into 5-6MPa and forms expanded solvents system, stirring and leaching 30-40min under room temperature, to normal pressure, again filters Pressure Drop to obtain filtrate 2; Merging filtrate 1 and filtrate 2, obtain extracting solution.
D, separation and purification
The extracting solution of step C gained being carried out to vacuum concentration, add petroleum ether dissolution, adopt silica gel column chromatography, is the petroleum ether-ethyl acetate wash-out of 3:1 (being that both volume ratio v/v are 3:1) by volume ratio, collects elutriant, and evaporation obtains crude product except desolventizing;
Crude product is carried out to separation with high speed adverse current chromatogram, solvent system is normal hexane-ethyl acetate-methanol-water, first the volume ratio with 16-20:1:20:1.2 is disposed at above-mentioned solvent system in separating funnel, shake up static layering, place 12-18h, getting upper strata is stationary phase, lower floor is moving phase, make sampling valve for sample introduction state, stationary phase is filled to chromatography column with pump, opening speed controller, make the chromatographic column forward of high-speed counter-current chromatograph, turn up 800r/min, get stationary phase and dissolve crude product to 20mg/ml, sample injection, flow rate of mobile phase is 2ml/min, according to detector ultraviolet spectrogram, receive Wuweizisu B component, vacuum concentration obtains Wuweizisu B sterling, and use HPLC to carry out purity check.
3. method according to claim 2, is characterized in that: the addition of sodium carbonate described in described steps A, for the 1-5% of dry Schisandra chinensis weight, is preferably 3%.
4. according to the method in claim 2 or 3, it is characterized in that: the ionic liquid in described step B is 1-butyl-2,3-methylimidazole chlorate, 1-butyl-2,3-methylimidazole Bromide, 1-butyl-2, any one in 3-methylimidazole tosilate.
5. according to the method described in claim 2-4 any one, it is characterized in that: the auxiliary agent in described step B is any one in diethylene glycol monomethyl ether, diethylene glycol dimethyl ether, dipropylene glycol monomethyl ether, dipropylene glycol dme.
6. according to the method described in claim 2-5 any one, it is characterized in that: the weight ratio of described step B intermediate ion liquid and auxiliary agent is 1:1-5, is preferably 1:3.
7. according to the method described in claim 2-6 any one, it is characterized in that: the 3-5% that in described step B, the weight of cetyl trimethylammonium bromide is ionic liquid, is preferably 4%.
8. according to the method described in claim 2-7 any one, it is characterized in that: in described step C, the weight ratio of Chinese Magnolivine Fruit and compound extractant is 1:6-10, is preferably 1:8.
9. according to the method described in claim 2-8 any one, it is characterized in that: in described step C, microwave extraction parameter is: microwave 350-550W, time 5-15min; Preferred microwave 400W, time 10min.
10. according to the method described in claim 2-9 any one, it is characterized in that: in described step C, the volume of expanded solvents consists of: 30% ethanol and 70%CO
2, the usage ratio of expanded solvents and Chinese Magnolivine Fruit is 2-4ml/g, is preferably 3ml/g.
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| CN104140909A (en) * | 2014-07-24 | 2014-11-12 | 安宁螳川源酒业经贸有限公司 | Rose wine and preparation method thereof |
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| CN101709059A (en) * | 2009-12-09 | 2010-05-19 | 北京博远欣绿科技有限公司 | Chinese magnoliavine fruit monomer composition separation preparation method |
| CN102050707A (en) * | 2009-11-01 | 2011-05-11 | 桂林莱茵生物科技股份有限公司 | Method for extracting deoxyschisandrin and schisandrin B from schisandra |
| CN103351373A (en) * | 2013-05-31 | 2013-10-16 | 浙江鼎辉医药科技有限公司 | Method for extracting Schisandrin B from Schisandra chinensis |
| CN103467438A (en) * | 2013-09-18 | 2013-12-25 | 浙江工业大学 | Method for extracting, separating and preparing lignin monomers from schisandra chinensis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102050707A (en) * | 2009-11-01 | 2011-05-11 | 桂林莱茵生物科技股份有限公司 | Method for extracting deoxyschisandrin and schisandrin B from schisandra |
| CN101709059A (en) * | 2009-12-09 | 2010-05-19 | 北京博远欣绿科技有限公司 | Chinese magnoliavine fruit monomer composition separation preparation method |
| CN103351373A (en) * | 2013-05-31 | 2013-10-16 | 浙江鼎辉医药科技有限公司 | Method for extracting Schisandrin B from Schisandra chinensis |
| CN103467438A (en) * | 2013-09-18 | 2013-12-25 | 浙江工业大学 | Method for extracting, separating and preparing lignin monomers from schisandra chinensis |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104140909A (en) * | 2014-07-24 | 2014-11-12 | 安宁螳川源酒业经贸有限公司 | Rose wine and preparation method thereof |
| CN104140909B (en) * | 2014-07-24 | 2015-08-19 | 安宁螳川源酒业经贸有限公司 | A kind of rose flower wine and preparation method thereof |
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