CN107714903A - The extraction technique of flavone compound in a kind of garlic - Google Patents
The extraction technique of flavone compound in a kind of garlic Download PDFInfo
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- CN107714903A CN107714903A CN201711192034.5A CN201711192034A CN107714903A CN 107714903 A CN107714903 A CN 107714903A CN 201711192034 A CN201711192034 A CN 201711192034A CN 107714903 A CN107714903 A CN 107714903A
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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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8962—Allium, e.g. garden onion, leek, garlic or chives
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- 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
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- 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
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- 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
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- 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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The present invention relates to a kind of extraction technique of flavone compound in garlic, garlic raw material is not required to dry and directly used, broken wall, complex enzyme zymohydrolysis, ultrasonic wave added ethonal extraction method are carried out using flash extracter in extraction process, the technology that broken wall and complex enzyme zymohydrolysis combination are carried out using flash extracter substantially increases extraction efficiency.
Description
Technical field
The present invention relates to a kind of extraction process, the extraction technique of flavone compound in more particularly to a kind of garlic.
Background technology
Garlic (English name Garlic；Latin name AlliumSativum), it is Liliaceae (Liliaceous) allium
(Allium) the Di Xia Scales stems of plant, there is 7~11 garlic cloves in garlic, narrow outer width in each garlic clove, white, plump succulence,
It is wrapped in film, around scape surrounding growth, so as to form oblate spheroid.
Garlic has unique healthcare function, there is many benefits to body, all such drug effects to be derived from it rich
Rich chemical composition, the toasted garlic of different sources have same drug effect and chemical composition, but due to geographical difference, chemical composition
Content it is different, the protein that is played a role in health care in people's garlic, amino acid, vitamin, sulfide equal size are also just slightly
Difference.Garlic is studied from German famous chemist special Dorr difficult to understand within 1844, the chemist then from Hua Jie various regions
The chemical composition and catabolite having one after another to garlic are studied.
The non-volatile compounds of garlic mainly include steroidal saponin, soap former times aglucon, flavonoid class, phenols, hemagglutinin,
Levulan and various amino acid etc..Mainly there is Huang Ugly class compounds in garlic：Kaempferol, cyanidenon, apiolin, red bayberry
Glycosides etc..
Flavone compound is plant some secondary metabolites caused by during long-term natural selection, is widely present
It is the fragility through a kind of active material caused by photosynthesis with regulation capillary in veterinary antibiotics and medicinal plant
It is immune and antitumor with permeability, protection cardiovascular system, anti-oxidant, elimination free radical, reduction blood glucose, anti-aging, enhancing
A variety of physiologically actives and the pharmacological action such as effect, anti-infective, antibacterial, antiviral and antiallergy.
The extracting method of flavones has a variety of, conventional methods to have circumfluence distillation, the extraction of alkali formula, surname extraction etc..Such as
What spring thunder etc. (what spring thunder, Luo Xueping, Li Lixia, research [J] the Sichuan Agricultural Universities journal of bamboo-leaves flavones extraction processes is waited,
2007,24(4):Bamboo-leaves flavones 409-412.) are extracted with heat reflow method, being determined by experiment optimal extract process is：Gu
Liquor ratio 1：20,80 DEG C of temperature, extraction time 30min.Now recovery rate is higher, and content is up to 13.7mg/g.Nearest 10 years, produce again
Supercritical ultrasonics technology, microwave method and supercritical CO 2 method extraction bamboo-leaves flavones are given birth to.Such as Qu Fenxia have studied multistage microwave amplifier leaf of bamboo Huang
Ketone, concentration of alcohol, leaf harvest time, solid-liquid ratio and these factors of Microwave Extraction time are probed into the shadow of extracting flavonoids amount
Ring, obtaining optimum extraction condition is：Collection in June blade, solid-liquid ratio 1: 30,60% concentration of alcohol, Microwave Extraction time 6min.
China Patent Publication No. is that CN102805798A discloses total Huang in a kind of extraction garlic bulb of one kind extraction garlic
The method of ketone.This method step is：Using garlic bulb as material, using distilled water as extractant, liquid ratio is chosen, ageing time and old
It is independent variable to change three factors of temperature, different levels is separately designed, using general flavone content as dependent variable, according to Box-
Benhnken models, meet with a response face experimental design, by carrying out burin-in process to garlic bulb, is greatly improved garlic bulb
The extraction efficiency of middle general flavone, effectively extract the Flavonoid substances in garlic bulb.
Extraction purification report in the prior art on flavones is existing a lot, but so far still without a kind of energy consumption of discovery
It is low, the time is short, process cleans, technique are simple and the method for good separating effect, therefore develop a kind of new and effective extracting flavonoids with
Purification technique is highly desirable.
The content of the invention
It is an object of the invention to provide a kind of extraction technique of flavone compound in garlic, extracted from garlic
General flavone, using breaking-wall cell and the technology that is combined of enzymolysis, have simple to operate, it is excellent to pollute few reagent recovery utilization rate height etc.
Point.
The extraction technique of flavone compound, its step are as follows in a kind of garlic：
1) fresh garlic crushes, and adds alcoholic solvent and soaks 1~3 hour, then carries out extraction using flash extracter,
Obtain broken wall slurries；
2) added in the shell-broken liquid pectase of shell-broken liquid weight 0.3%, 1.0% cutinase composition it is compound
Enzyme, in the basic conditions after 5~6h of enzyme, enzymolysis liquid is obtained after destroy the enzyme treatment；
3) enzymolysis liquid is homogenized addition alcoholic solvent ultrasonic extraction and obtains general flavone extract solution twice；
4) concentrated extracting solution upper macroreticular resin, is washed till clarification of water with water, 5~10% ethanol successively, finally used to paste
80-85% ethanol elutions, eluent remove ethanol and obtain concentrate A；
5) by concentrate A with being filtered after alcohol solvent, filtrate mixes sample, upper polyamide chromatography post, using dichloro with polyamide
Hexane：Methanol is 10:1~1:1 ratio carries out gradient elution, and the eluent containing general flavone, eluent are collected according to TLC points version
Concentrate B is concentrated to give, then flavones is obtained after concentrate B is freeze-dried.
Flash extracter described in step 1) is a kind of high and new technology equipment for Chinese herbal medicine extraction, and the equipment is to organize
Based on smashing principle, according to the purpose and process characteristic, technical requirements etc. of extraction, foreign latest technology, science are fully absorbed
Design, is elaborated, and has not only reached quick, and efficiently, fully, complete extraction requires, and is not required to heat, and saves the energy,
The heat-sensitive ingredients in extraction of substance are effectively protected, needed the extraction work of a few hours originally only needs a few minutes using the instrument
It can complete, operating efficiency improves tens times, and easy to operate, excellent with highly significant compared with traditional extraction process
Gesture, the strictly handy extraction equipment of Chinese medicine researcher.
Flash extracter is crushed extraction cutter head and control system three parts forms, decoration structure features simple design, only by high-speed electric expreess locomotive
Gather, it is rationally, easy to operation.As long as the part of its work is broken extraction cutter head, cutter head is made up of interior knife and outer knife, interior during work
Knife adjusts its speed under the drive of high-speed electric expreess locomotive by control system.Rotated at a high speed in outer knife intracavitary, whole system is in one
Among the individual dynamic environment of high speed, powerful shear action is not only produced between inside and outside knife, while the generation of outer knife intracavitary is powerful
Negative pressure, in the presence of this negative pressure, outer knife intracavitary outgoing sub- phenomenon of osmosis estranged, both fully exposed flavonoids by broken
Material is surrounded under the various external force effects such as negative pressure, shearing, high velocity impact by solvent molecule, dissociates, dissolves, substitute, departs from, so
After quickly enter in solvent, moment reaches the balance of solvent strength, and extraction process is rapidly completed within the several seconds.This process is abundant
Absorb and it is ingenious combine shearing, vacuum, permeate, the principle such as fluid dynamic, realize and combine the effect mutually held between technology.
One or more of the alcoholic solvent in methanol, ethanol, propyl alcohol, butanol in step 1).The homogenate extraction voltage
For 220V, 1~3 minute extraction time.
Step 2) neutral and alkali condition is that pH value is 10~12.Described cutinase can be any source, including lactation is moved
Thing, plant or microbe-derived, but preferred microorganism source.Specifically described cutinase is derived from filamentous fungi or yeast
Cutinase.
The concrete operations of homogenate are homogenized 3~10min under the conditions of being 3000~6000r/min in step 3).
Macroreticular resin is DiaionHP-10 types, SPD100, D-101 type or AB-8 types in step 4).
It is an advantage of the invention that：
(1) technology being combined using flash extracter broken wall treatment at complex enzyme, the cell of cell is effectively destroyed
Wall, the mass transfer area that Flavonoid substances during subsequent extracted spread to ethanol is increased, significantly improves flavone compound
Recovery rate；
(2) homogenized is carried out after the Flavonoid substances after complex enzyme hydrolysis is handled mix with ethanol, it is ensured that flavones with
Alcohol solvent is sufficiently mixed, and in the presence of high shear is homogenized, and cell occurs further to crush, promote effectively into
Divide the extraction of flavones.
Embodiment
The technical scheme in the embodiment of the present invention is clearly and completely described below, it is clear that described embodiment
Only part of the embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, the common skill in this area
The every other embodiment that art personnel are obtained on the premise of creative work is not made, belong to the model that the present invention protects
Enclose.
【Embodiment 1】
1) take 1kg fresh garlics to crush, add 2L ethanol and soak 1 hour；Using flash extracter in the case where voltage is 220V
Carry out broken wall treatment 3 minutes, obtain broken wall slurries.
2) 0.3% pectase, the complex enzyme of 1.0% cutinase composition, and pH is adjusted as 11, after digesting 5 hours, height
Warm destroy the enzyme treatment obtains enzymolysis liquid.
3) enzymolysis liquid is homogenized addition alcoholic solvent ultrasonic extraction and obtains general flavone extract solution twice；
4) concentrated extracting solution upper macroreticular resin, is washed till clarification of water with water, 5~10% ethanol successively, finally used to paste
80-85% ethanol elutions, eluent ethanol of going out obtain concentrate A；
5) by concentrate A with being filtered after alcohol solvent, filtrate mixes sample, upper polyamide chromatography post, using dichloro with polyamide
Hexane：Methanol is 10:1~1:1 ratio carries out gradient elution, and the eluent containing general flavone, eluent are collected according to TLC points version
Concentrate B is concentrated to give, then flavones 1.02g is obtained after concentrate B is freeze-dried, is detected through HPLC, product purity contains for flavones
Amount 96.3%.
【Embodiment 2】
1) take 1kg fresh garlics to crush, add 2L ethanol and soak 1 hour；Using flash extracter in the case where voltage is 220V
Carry out broken wall treatment 3 minutes, obtain broken wall slurries.
2) 0.3% pectase, the complex enzyme of 1.0% cutinase composition, and pH is adjusted as 11, after digesting 5 hours, height
Warm destroy the enzyme treatment obtains enzymolysis liquid.
3) enzymolysis liquid is homogenized addition alcoholic solvent ultrasonic extraction and obtains general flavone extract solution twice；
4) concentrated extracting solution upper macroreticular resin, is washed till clarification of water with water, 5~10% ethanol successively, finally used to paste
80-85% ethanol elutions, eluent ethanol of going out obtain concentrate A；
5) by concentrate A with being filtered after alcohol solvent, filtrate mixes sample, upper polyamide chromatography post, using dichloro with polyamide
Hexane：Methanol is 10:1~1:1 ratio carries out gradient elution, and the eluent containing general flavone, eluent are collected according to TLC points version
Concentrate B is concentrated to give, then flavones 0.97g is obtained after concentrate B is freeze-dried, is detected through HPLC, product purity contains for flavones
Amount 97.1%.
【Comparative example 1】
1) take 1kg fresh garlics to crush, add 2L ethanol and soak 1 hour；Using flash extracter in the case where voltage is 220V
Carry out broken wall treatment 3 minutes, obtain broken wall slurries.
2) broken wall slurries are homogenized addition alcoholic solvent ultrasonic extraction and obtain general flavone extract solution twice；
3) concentrated extracting solution upper macroreticular resin, is washed till clarification of water with water, 5~10% ethanol successively, finally used to paste
80-85% ethanol elutions, eluent ethanol of going out obtain concentrate A；
4) by concentrate A with being filtered after alcohol solvent, filtrate mixes sample, upper polyamide chromatography post, using dichloro with polyamide
Hexane：Methanol is 10:1~1:1 ratio carries out gradient elution, and the eluent containing general flavone, eluent are collected according to TLC points version
Concentrate B is concentrated to give, then flavones 0.56g is obtained after concentrate B is freeze-dried, is detected through HPLC, product purity contains for flavones
Amount 93.7%.
The extraction technique of flavone compound, has the following advantages that in a kind of garlic of the present invention：
(1) technology being combined using flash extracter broken wall treatment at complex enzyme, the cell of cell is effectively destroyed
Wall, the mass transfer area that Flavonoid substances during subsequent extracted spread to ethanol is increased, significantly improves flavone compound
Recovery rate；
(2) homogenized is carried out after the Flavonoid substances after complex enzyme hydrolysis is handled mix with ethanol, it is ensured that flavones with
Alcohol solvent is sufficiently mixed, and in the presence of high shear is homogenized, and cell occurs further to crush, promote effectively into
Divide the extraction of flavones.
Described above has fully disclosed the embodiment of the present invention.It is pointed out that it is familiar with the field
Scope of the technical staff to any change that the embodiment of the present invention is done all without departing from claims of the present invention.
Correspondingly, the scope of claim of the invention is also not limited only to previous embodiment.
Claims (8)
1. the extraction technique of flavone compound, its step are as follows in a kind of garlic：
1) fresh garlic crushes, and adds alcoholic solvent and soaks 1~3 hour, then carries out extraction using flash extracter, obtain
Broken wall slurries；
2) pectase, the complex enzyme of 1.0% cutinase composition of shell-broken liquid weight 0.3% are added in the shell-broken liquid,
Under alkalescence condition after 5~6h of enzyme, enzymolysis liquid is obtained after destroy the enzyme treatment；
3) enzymolysis liquid is homogenized addition alcoholic solvent ultrasonic extraction and obtains general flavone extract solution twice；
4) concentrated extracting solution upper macroreticular resin, is washed till clarification of water with water, 5~10% ethanol successively, finally uses 80- to paste
85% ethanol elution, eluent remove ethanol and obtain concentrate A；
5) by concentrate A with being filtered after alcohol solvent, filtrate mixes sample, upper polyamide chromatography post, using dichloro hexane with polyamide：
Methanol is 10:1~1:1 ratio carries out gradient elution, and the eluent containing general flavone, eluent concentration are collected according to TLC points version
Concentrate B is obtained, then flavones will be obtained after concentrate B freeze-dryings.
2. the extraction technique of flavone compound in garlic according to claim 1, it is characterised in that in step 1)
One or more of the alcoholic solvent in methanol, ethanol, propyl alcohol, butanol.
3. the extraction technique of flavone compound in garlic according to claim 2, it is characterised in that the alcohol is molten
Agent is ethanol.
4. the extraction technique of flavone compound in garlic according to claim 1, it is characterised in that in step 1)
Homogenate extraction voltage is 220V, 1~3 minute extraction time.
5. the extraction technique of flavone compound in garlic according to claim 1, it is characterised in that in step 2)
Alkalescence condition is that pH value is 10~12.
6. the extraction technique of flavone compound in garlic according to claim 1, it is characterised in that in step 2)
Described cutinase can be any source, including mammal, plant or microbe-derived, but preferred microorganism source.Tool
Described cutinase is derived from the cutinase of filamentous fungi or yeast for body.
7. the extraction technique of flavone compound in garlic according to claim 1, it is characterised in that in step 3)
The concrete operations of homogenate are homogenized 3~10min under the conditions of being 3000~6000r/min.
8. the extraction technique of flavone compound in garlic according to claim 1, it is characterised in that in step 4)
Macroreticular resin is DiaionHP-10 types, SPD100, D-101 type or AB-8 types.
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| CN201711192034.5A CN107714903B (en) | 2017-11-24 | 2017-11-24 | Wall-breaking extraction process of flavonoid compounds in garlic |
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| CN201711192034.5A CN107714903B (en) | 2017-11-24 | 2017-11-24 | Wall-breaking extraction process of flavonoid compounds in garlic |
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| CN107714903A true CN107714903A (en) | 2018-02-23 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108719930A (en) * | 2018-05-25 | 2018-11-02 | 金华金字火腿有限公司 | A kind of high efficiency preparation method of Jinhua ham bone taste compound |
| CN110231296A (en) * | 2019-07-16 | 2019-09-13 | 郑州轻工业学院 | A kind of quick green high-efficient extracts the method and its detection method of silkworm excrement flavones |
| CN112402482A (en) * | 2020-12-07 | 2021-02-26 | 北京美康堂医药科技有限公司 | Method for extracting and purifying flavonoid components in traditional Chinese medicinal materials |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102805798A (en) * | 2012-08-23 | 2012-12-05 | 西北农林科技大学 | Method for extracting general flavone from garlic bulbs |
-
2017
- 2017-11-24 CN CN201711192034.5A patent/CN107714903B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102805798A (en) * | 2012-08-23 | 2012-12-05 | 西北农林科技大学 | Method for extracting general flavone from garlic bulbs |
Non-Patent Citations (2)
| Title |
|---|
| 李利华: "大蒜总黄酮的超声波辅助提取及其抗氧化活性", 《食品与发酵工业》 * |
| 李筱玲等: "黄酮类化合物提取分离方法研究进展", 《陕西农业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108719930A (en) * | 2018-05-25 | 2018-11-02 | 金华金字火腿有限公司 | A kind of high efficiency preparation method of Jinhua ham bone taste compound |
| CN110231296A (en) * | 2019-07-16 | 2019-09-13 | 郑州轻工业学院 | A kind of quick green high-efficient extracts the method and its detection method of silkworm excrement flavones |
| CN110231296B (en) * | 2019-07-16 | 2021-09-17 | 郑州轻工业学院 | Method for rapidly, greenly and efficiently extracting silkworm excrement flavone and detection method thereof |
| CN112402482A (en) * | 2020-12-07 | 2021-02-26 | 北京美康堂医药科技有限公司 | Method for extracting and purifying flavonoid components in traditional Chinese medicinal materials |
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Effective date of registration: 20221201 Address after: 710076 Room 30606, Building B3, Jinye Times, No. 32, Jinye Road, High tech Zone, Xi'an, Shaanxi Patentee after: Xi'an Aogu Biotechnology Co.,Ltd. Address before: 310052 Room 805, 8th Floor, Building 3, No. 487, Jianghui Road, Changhe Street, Binjiang District, Hangzhou, Zhejiang Patentee before: HANGZHOU GENGLAN BIOLOGICAL TECHNOLOGY Co.,Ltd. |