CN107998172A - Fat-soluble flavones microwave radiation technology three-phase separate matches somebody with somebody extracting method in a kind of blueberry leaf - Google Patents
Fat-soluble flavones microwave radiation technology three-phase separate matches somebody with somebody extracting method in a kind of blueberry leaf Download PDFInfo
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- CN107998172A CN107998172A CN201711347699.9A CN201711347699A CN107998172A CN 107998172 A CN107998172 A CN 107998172A CN 201711347699 A CN201711347699 A CN 201711347699A CN 107998172 A CN107998172 A CN 107998172A
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- fat
- extracting method
- flavones
- microwave
- blueberry leaf
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/45—Ericaceae or Vacciniaceae (Heath or Blueberry family), e.g. blueberry, cranberry or bilberry
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
Abstract
The invention discloses fat-soluble flavones three-phase separate in a kind of blueberry leaf to match somebody with somebody extracting method, includes the following steps：1) grind into powder after blueberry leaf is dried；2) powder obtained in step 1) is mixed with water, then adds ammonium sulfate and the tert-butyl alcohol thereto, be stirred；3) microwave irradiation is carried out to the mixed system of step 2), to form three-phase layer, and records three-phase layer and form the required time；4) organic phase is separated, fat-soluble flavones is prepared in further separation.It is combined using three-phase distribution method with microwave loss mechanisms from blueberry leaf and extracts fat-soluble flavones, the extract yield of flavones can be greatly improved, and extraction time can be shortened, simplifies extraction process.
Description
Technical field
A kind of extractive technique field of active ingredient in blueberry leaf of the present invention, and in particular to fat-soluble flavones three in blueberry leaf
Phase partitioning extracting method.
Background technology
Blueberry (Semen Trigonellae) means the meaning of the berry of blueness, also known as：Lan Mei, bog bilberry, sincere persimmon, beep is thermophilic, all
Persimmon, pasture fruit, great bilberry, belongs to perennial fallen leaves or evergreen shrubs fruit tree, in shrub, fruit is berry.Except taste of fruit is unique
Full of nutrition outer, blueberry leaf can also be used as medicine.Contain crude protein, crude fibre, aliphatic acid, sterol, terpene, mineral member in blueberry leaf
Element, amino acid, organic acid, vitamin, carbohydrate and a large amount of flavone compounds, the effect of making it have diuresis, anti-inflammatory, detoxify,
And available for rheumatism, gout etc..Flavones therein is acted on stronger scavenging activated oxygen, and anti-lipid peroxidation, is one
Kind high activity, the natural materials having no toxic side effect.
At present, it is soxhlet extraction or microwave loss mechanisms that the extracting method of Flavonoid substances is common in blueberry leaf, Soxhlet
Extraction method is that powdery blueberry leaf is placed in casing rack, and uses pure solvent as Extraction solvent.Solid matter is placed on filter paper
In set, it is positioned in extraction cell, after solvent ebuillition of heated, steam is risen by gas-guide tube, is condensed into liquid and is instilled extraction
In device.When liquid level exceedes siphon pipe highest point, that is, siphonage, solution reflux such as flask, therefore can extract and be dissolved in occurs
The material of solvent, using solvent refluxing and siphonage, makes the soluble matter in solid be enriched in flask.This kind of extracting method needs
More solvent is used, extraction time and the energy consumption needed are more, and the extract yield of Flavonoid substances is relatively low.Microwave carries
It is that blueberry leaf sample is positioned in organic extraction solvent to extract to follow the example of, and by extraction system be positioned in Microwave Extraction Equipment into
Row microwave irradiation, to accelerate the extraction of Flavonoid substances, Extraction solvent is generally methanol, ethanol, acetone or ethyl acetate, this kind
The usage amount of the Extraction solvent of method is also larger, and the extract yield of Flavonoid substances is relatively low.
The content of the invention
For above-mentioned technical problem existing in the prior art, the object of the present invention is to provide fat-soluble in a kind of blueberry leaf
Flavones three-phase separate matches somebody with somebody extracting method, which can not only reduce the usage amount of organic solvent, simplify extraction process, may be used also
To improve the extract yield of fat-soluble flavones.
In order to solve problem above, the technical scheme is that：
Fat-soluble flavones microwave radiation technology three-phase separate matches somebody with somebody extracting method in a kind of blueberry leaf, includes the following steps：
1) grind into powder after blueberry leaf is dried；
2) powder obtained in step 1) is mixed with water, then adds ammonium sulfate and the tert-butyl alcohol thereto, be stirred；
3) to step 2) mixed system carry out microwave irradiation, to form three-phase layer, and record three-phase layer formed it is required
Time；
4) organic phase is separated, fat-soluble flavones is prepared in further separation.
It is combined using three-phase distribution method with microwave loss mechanisms from blueberry leaf and extracts fat-soluble flavones, can be greatly improved
The extract yield of flavones, and extraction time can be shortened, simplify extraction process.
Preferably, in step 1), the dried moisture of blueberry leaf is 8-10%.Preserved in blueberry leaf to be extracted suitable
Work as moisture, be conducive to improve the extract yield of flavones.
Preferably, in step 1), the particle diameter for the powder that blueberry leaf is ground into is 0.5-1mm.
Preferably, in step 2), the amount of the ammonium sulfate, water and the tert-butyl alcohol that are added in every gram of powder is respectively 6-7g, 35-
45ml、35-45ml。
It is further preferred that in step 2), the volume ratio of water and the tert-butyl alcohol is 1:1.
Preferably, in step 2), the ammonium sulfate of addition is the aqueous solution of ammonium sulfate, and concentration 30%-60%, % are quality
Percentage, is preferably 40%.
Preferably, in step 3), the power of microwave irradiation is 250-300W, is preferably 280W.
It is further preferred that in step 3), the time of microwave irradiation is 3-10min, is preferably 5min.
It is further preferred that the microwave irradiation irradiates for pulse mode.Microwave irradiation is divided into pulse mode and the progressive die
Formula, when continuous mode irradiates, can make system persistently overheating, may make degradation.And under pulse mode, intervals of power supplies
Should, the chance of degradation is relatively fewer, is conducive to improve the quality of extract.But also can be to avoid unnecessary energy
Use, avoid the evaporation and boiling of solvent.
Still more preferably, the duty cycle of microwave irradiation is 25%-50%, is preferably 50%.
Preferably, in step 3), the temperature of extraction is 28-32 DEG C.
Beneficial effects of the present invention are：
1) it is combined using three-phase distribution method with microwave loss mechanisms from blueberry leaf and extracts fat-soluble flavones, can be carried significantly
The extract yield of high flavones, and extraction time can be shortened, simplify extraction process.
2) power of microwave irradiation is one of important parameter for influencing extract yield, when the power of microwave irradiation is 250-
During 300W, good extract yield can be obtained.
Embodiment
It is noted that described further below is all illustrative, it is intended to provides further instruction to the application.It is unless another
Indicate, all technical and scientific terms used herein has usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe embodiment, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " bag
Include " when, it indicates existing characteristics, step, operation, device, component and/or combinations thereof.
Match somebody with somebody to be combined from blueberry leaf with microwave using three-phase separate and extract fat-soluble flavones, 50mg/g is obtained in 5h.Most
Under the conditions of good, such as microwave irradiation time 5min, ammonium sulfate concentrations 40%w/v, power 280W, the ratio of solute and solvent is 1:
20, the ratio of slurries and the tert-butyl alcohol is 1:1, soaking time 5min, by microwave radiation technology three-phase separate with the fat-soluble of extraction acquisition
Yield of flavone is 50mg/g.The extracting method developed generates higher extract yield in the short period of time.
Material
The moisture of blueberry climax leaves powder is about 9-10%, particle size range 0.5-1mm.The moisture of powder is about
For 9%-10%, particle size range 0.5-1mm.Initially by leaf clean and dry 48 it is small when.Then the crushing up leaves dried are simultaneously
It is stored in the sealing container in refrigerator.
Method
All experiments carry out at a temperature of 30 ± 2 DEG C
1st, the method that microwave radiation technology three-phase separate is matched somebody with somebody
Specific steps；
1) leaf powder and water initially will be prepared with 1：In 40 ratio microwave abstracting glass containers, slurries are made.
2) ammonium sulfate is added in slurry fractions, concentration is reached 30%w/v.
3) under the action of intermediate agitation, then add and the isometric tert-butyl alcohol of slurry.
4) by the mixture through microwave irradiation, to form three layers of phase in microwave.
5) record forms three layers of mutually required time.
6) hold the mixture in separatory funnel to separate organic layer and water layer.
7) organic phase containing the tert-butyl alcohol is analyzed to determine fat-soluble flavones content.
In order to study the effect for the various parameters for using microwave radiation technology three-phase separate to match somebody with somebody, similar experiment has been carried out.It has studied
Microwave irradiation time, ammonium sulfate concentrations (30-60%w/v), microwave input power (150,280,400W), slurries and tert-butyl alcohol ratio
Example (1：0.5-1：4), solute (1：10-1：60), soaking time (2-20 minutes) and working cycles number (6) are to fat-soluble flavones
The influence of extraction.
2nd, condition optimizing
Influence of the microwave time to fat-soluble flavones
Microwave irradiation time increases to 7 minutes from 0 causes increasing sharply for fat-soluble extracting flavonoids rate.Microwave irradiation time
For 5min when, yield highest.Target compound increases above 5 minutes with microwave irradiating time, causes with the charing of material
Evaporation of the solvent, causes yield to reduce.Therefore 5 minutes deemed appropriate microwave irradiating times,
Influence of the work period to fat-soluble flavones
Micro-wave oven can work under pulse mode or continuous mode.It is not recommended that operate in a continuous mode, because it can lead
Cause continuous molecular friction.This can continue to increase temperature, this may make degradation.And in the pulsing mode, between power
Every supply, therefore the chance of degradation is relatively fewer.In addition, operation will avoid unnecessary energy from making in a pulsed mode
With, and can be to avoid the evaporation and boiling of solvent.25,50,65,80 and 100% duty cycle under assess duty cycle to fat
The influence of dissolubility extracting flavonoids rate.The yield highest at 50% duty cycle (close for 1 minute and 1 minute opens).More than 25%
Work period in, yield is kept approximately constant.In order to save energy, preferably operated under relatively low and optimal duty cycle micro-
Ripple extracts, therefore 50% duty cycle is used to further test.
Influence of the ammonium sulfate concentrations to fat-soluble flavones
By maintaining other experiment conditions constant, solute is 1 with solvent ratios:40, microwave irradiating time 5 minutes and slurries
It is 1 with the ratio between the tert-butyl alcohol：1, tested under 30% to 60%w/v ammonium sulfate concentrations.Ammonium sulfate has in 40%w/v
Maximum extraction efficiency.This is probably the dehydration due to sulfate ion, changes the solubility of fat-soluble flavones, causes
The unfavorable conditions of fat-soluble flavones in water phase, so as to promote fat-soluble flavones to enter butanol phase.Therefore, the sulfuric acid of 40%w/v
Ammonium is considered as the optimization concentration of next experiment.
Influence of the power to fat-soluble flavones
Microwave input power is to determine one of yield and the important parameter of extraction time, and the main original of processing cost
Cause.If power input is excessive, evaporation of the solvent can be caused, cause plant substrates to carbonize.The influence of power input is 150,280
Studied with 400W.Remaining experiment condition is：The ratio of solute and solvent is 1:40, microwave irradiating time is 5 minutes,
The ratio of slurries and the tert-butyl alcohol is 1：1, ammonium sulfate concentrations are 40% (w/v).Higher recovery rate is obtained in 280W.Most
Just, the increase of microwave power input causes the increase of power consumption, so as to add the yield of fat-soluble flavones.It is moreover, more
Electromagnetic energy be passed, and power provide sample in local heating.This destroys plant substrates in a short time molten
Solute is spread in agent.The reduction of yield is attributed to high-power the fact that dissipate with smaller size smaller under higher microwave power, this is short
The temperature of mixture is increased in time and causes the charing of particle.Very high microwave radiation provides for solvent and matrix
Unnecessary energy, this understands the interaction between disturbing molecule so as to influence extraction process.
Influence of the tert-butyl alcohol ratio to fat-soluble flavones in slurries
The influence of slurry and tert-butyl alcohol ratio to fat-soluble extracting flavonoids effect is have studied, is changed into 1：0.5,1：1,
1：2 and 1：4.Previous experiments parameter such as solute/solvent is than 1:40, microwave irradiating time 5min, power input 280W, ammonium sulfate are dense
40% (w/v) of degree is kept constant.When ratio is 1：1, recovery rate also remains unchanged, and further with slurries ratio reduces.Uncle
The increase of the amount of butanol will not increase yield, because the increase of the amount with the tert-butyl alcohol, the viscosity increase of slurry.With viscosity
Increase, the mobility of molecule decline so that molecule is difficult to be aligned with microwave field.Which reduce the absorption of microwave energy, so as to subtract
The heat that dipole rotation produces is lacked.Since the amount of the tert-butyl alcohol is not enough to be separated, the tert-butyl alcohol cannot have with ammonium sulfate
The cooperative effect of effect, so the ratio of the tert-butyl alcohol is less than 1 in slurries：3.Therefore the ratio of suitable slurry and the tert-butyl alcohol should be kept
Example (1：1) to obtain appropriate extraction.
Influence of the solute/solvent ratio to fat-soluble flavones
The ratio of solute and solvent is to determine one of solvent acquisition and the important parameter of Downstream processing cost.It should keep flat
Weigh to obtain optimal yield and avoid unnecessary solvent from using.Solute ratio is 1:10-1:50, microwave irradiation time 5min, sulphur
Sour ammonium concentration 40%w/v, input power 280W, slurry and the tert-butyl alcohol 1：1 mixing.In fat-soluble flavones yield in solute/solvent
Ratio 1:Highest when 20, then declines.This is because significant absorption of the polar solvent to microwave radiation only results in less wear
Saturating depth.Therefore, when adding a large amount of solvents, it may occur that the excessive expansion of material.Therefore, relatively small number of microwave energy is direct
It is absorbed by material, because microwave is completely absorbed on border.
The influence that soaking time matches somebody with somebody fat-soluble flavones microwave radiation technology three-phase separate
Soaked respectively 0,2,5,10 and 20 minute with the 0.5g powder being immersed in 20mL water, research soaking time is to liposoluble
The influence of property extracting flavonoids.Under the power of immersion known time, 40%w/v ammonium sulfate is added, then adds 1：1 tert-butyl alcohol.
Then microwave treatment is carried out 5 minutes under conditions of power input is 280W.Higher carry is obtained when soaking time is 5 minutes
Take.When dry powder soaks 5min, fat-soluble flavones yield increases to 12.5% or so.Since water is best absorption microwave material
Material, therefore the content that can moisturize is soaked, so as to cause the microwave penetration of higher.Increased microwave penetration makes cell more effectively
Rupture, discharges solute, so as to improve recovery rate.Therefore, optimization soaking time will be helpful to realize the optimum extraction of biomolecule.
It is that the present invention uses various parameters optimal when extracting fat-soluble flavones below：
1) 5 minutes deemed appropriate microwave irradiating times；
2) extraction efficiency of the ammonium sulfate in 40%w/v with maximum；
3) higher recovery rate is obtained in 280W；
4) tert-butyl alcohol ratio 1 in slurries：1 recovery rate highest；
5) fat-soluble flavones yield is in solute/solvent ratio 1:Highest when 20；
6) higher extraction is obtained when soaking time is 5 minutes.
3rd, the method control of surname extraction：
Surname extraction is used as the reference method compared with fat-soluble flavones.When extraction progress 5 is small.Fat-soluble flavones carries
It is 52mg/g to take rate.
Compared with soxhlet extraction, the advantages of microwave radiation technology three-phase distribution method, is：
1) microwave radiation exaraction needs less solvent, time and energy.
2) extraction process is not only simplify, and recovery rate and soxhlet extraction method are relatively.Yield difference is little, but when
Between greatly save.
3) it is fast and effective simple, improve efficiency.
4) it is used as previous step before what Downstream processing, and can be also used for extraction/separation compound.
As a result and discuss
The fat-soluble flavones yield that this method obtains is about 50mg/g in 5 minutes under 280W input powers.It is fat-soluble
The yield of flavones is the 95% of soxhlet extraction (52mg/g), and the time is extremely short (fat-soluble flavones 5min, soxhlet extraction 5h).
This method causes to extract and separate at the same time by overcoming the mass transfer and limit related with Microwave Extraction.For extracting fat-soluble flavones
It is proved to be highly effective and quick technology, it can be used as previous step before any Downstream processing, and may be used also
For extraction/separation compound.
The comparison of ultrasonic wave assisted extraction and three-phase distribution method
The fat-soluble flavones yield obtained in ultrasonic wave assisted extraction in 25 minutes is 41mg/g, and three-phase distribution method
Produce 50mg/g.Due to many reasons, the productivity ratio ultrasonic wave assisted extraction that three-phase distribution method obtains is high.First, leaf powder contains
A certain amount of moisture.Therefore, when with microwave irradiation slurry, moisture absorption energy present in particle.As a result, cell wall expansion
And rupture, release component enters solvent.Once compound is extracted in solvent, the synergistic effect of microwave and the dehydration of ammonium sulfate
Fat-soluble flavones is just transferred in tert-butyl alcohol phase rapidly by effect, while extracts and purify fat-soluble flavones.Secondly, see in an experiment
Observe, compared with ultrasonic wave, microwave power consumption is less.Therefore, compared with supersound process, the energy of higher is by liposoluble in microwave
Property flavones is quickly pushed into butanol phase, so as to help to extract and separate.
The foregoing is merely the preferred embodiment of the application, the application is not limited to, for the skill of this area
For art personnel, the application can have various modifications and variations.It is all within spirit herein and principle, made any repair
Change, equivalent substitution, improvement etc., should be included within the protection domain of the application.
Claims (10)
1. fat-soluble flavones three-phase separate matches somebody with somebody extracting method in a kind of blueberry leaf, it is characterised in that：Include the following steps：
1) grind into powder after blueberry leaf is dried；
2) powder obtained in step 1) is mixed with water, then adds ammonium sulfate and the tert-butyl alcohol thereto, be stirred；
3) microwave irradiation is carried out to the mixed system of step 2), to form three-phase layer, and records three-phase layer and form the required time；
4) organic phase is separated, fat-soluble flavones is prepared in further separation.
2. extracting method according to claim 1, it is characterised in that：In step 1), the dried moisture of blueberry leaf
For 8-10%.
3. extracting method according to claim 1, it is characterised in that：In step 1), the grain for the powder that blueberry leaf is ground into
Footpath is 0.5-1mm.
4. extracting method according to claim 1, it is characterised in that：In step 2), the ammonium sulfate that is added in every gram of powder,
The amount of water and the tert-butyl alcohol is respectively 6-7g, 35-45ml, 35-45ml.
5. extracting method according to claim 4, it is characterised in that：In step 2), the volume ratio of water and the tert-butyl alcohol is 1:
1。
6. extracting method according to claim 1, it is characterised in that：In step 2), the ammonium sulfate of addition is ammonium sulfate
Aqueous solution, concentration 30%-60%, % are mass percent, are preferably 40%.
7. extracting method according to claim 1, it is characterised in that：In step 3), the power of microwave irradiation is 250-
300W, is preferably 280W.
8. extracting method according to claim 1, it is characterised in that：In step 3), the time of microwave irradiation is 3-
10min, is preferably 5min.
9. extracting method according to claim 8, it is characterised in that：The microwave irradiation irradiates for pulse mode；Microwave
The duty cycle of irradiation is 25%-50%, is preferably 50%.
10. extracting method according to claim 1, it is characterised in that：In step 3), the temperature of extraction is 28-32 DEG C.
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| CN201711347699.9A CN107998172A (en) | 2017-12-15 | 2017-12-15 | Fat-soluble flavones microwave radiation technology three-phase separate matches somebody with somebody extracting method in a kind of blueberry leaf |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711347699.9A CN107998172A (en) | 2017-12-15 | 2017-12-15 | Fat-soluble flavones microwave radiation technology three-phase separate matches somebody with somebody extracting method in a kind of blueberry leaf |
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2017
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|---|---|---|---|---|
| CN1391938A (en) * | 2002-07-30 | 2003-01-22 | 天津大学 | Microwave extracting method for Chinese medicine effective components |
| JP2005001998A (en) * | 2003-06-09 | 2005-01-06 | Api Co Ltd | Hypotensive agent, method for producing the same and propolis composition and food formulation |
| CN101290277A (en) * | 2007-04-20 | 2008-10-22 | 财团法人食品工业发展研究所 | Rapid extraction method utilizing microwave radiation |
| CN103432193A (en) * | 2013-09-10 | 2013-12-11 | 武汉工程大学 | Microwave-assisted aqueous two-phase extraction and separation method of kudzu root total flavones |
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| 李颖畅 等: "蓝莓叶总黄酮的提取及其定性分析", 《食品与发酵科技》 * |
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