CN109010504A - A kind of extracting method of lycium ruthenicum general flavone - Google Patents
A kind of extracting method of lycium ruthenicum general flavone Download PDFInfo
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- CN109010504A CN109010504A CN201811028506.8A CN201811028506A CN109010504A CN 109010504 A CN109010504 A CN 109010504A CN 201811028506 A CN201811028506 A CN 201811028506A CN 109010504 A CN109010504 A CN 109010504A
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- lycium ruthenicum
- general flavone
- extracting
- extracting method
- micro mist
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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)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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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
-
- 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
-
- 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
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- 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
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention belongs to the separation and extraction technologies of biology and the natural products effective ingredient of light industry and chemical industry biotechnology in new medicine, are related to plant extracts field, specially a kind of extracting method of lycium ruthenicum general flavone.After this method uses superfine communication technique that lycium ruthenicum micro mist is prepared first, micro mist is handled using microwave；Then the general flavone in lycium ruthenicum micro mist is extracted using the ionic liquid of ultrasonic wave added-salt double-aqueous phase system；Ultrafiltration is finally successively used, column chromatogram and supercritical fluid extraction purify lycium ruthenicum general flavone.The lycium ruthenicum general flavone purity is high of this method preparation, antibacterial action is obvious, can be directly as the functional additive of health care of food product.
Description
Technical field
The invention belongs to the separation of biology and light industry in new medicine and the natural products effective ingredient of chemical industry biotechnology to mention
Technology is taken, the extracting method that plant extracts field is specially a kind of lycium ruthenicum general flavone is related to.
Background technique
Lycium ruthenicum is a species for Solanaceae Lycium, has quil, and berry is spherical, thin skin, atropurpureus after skin is ripe.Lycium ruthenicum
Sweet succulence, vitamin, organic acid and carbohydrate rich in contain anthocyanin and flavone compound in fruit.Flavones conduct
One of main component of lycium ruthenicum have anti-oxidant, anti-aging, antiatherosclerosis, reducing blood lipid, it is antifatigue, hypoglycemic and
The effects of enhancing non-specific immune function.Fructus lycii used in food is mostly full fructus lycii and fructus lycii powder, consumer's food at present
It is difficult to completely absorb active constituent therein after, so that fructus lycii product health-care effect is unobvious.Therefore exploitation one
It plants the extracting method of lycium ruthenicum general flavone and extract will be obtained and have applied in food to the health-care effect for improving fructus lycii product
There is important meaning.
The extracting method of general flavone is mostly and has in plant more rare about the document report of lycium ruthenicum at present and traditional
Solvent extraction or ultrasonic wave assisted extraction method, but obtained product extraction efficiency is not high and while purifying uses tradition more
Liquid-liquid extraction purified, organic solvent consumption it is big and time-consuming and laborious, therefore develop high-efficiency environment friendly lycium ruthenicum general flavone mention
Method is taken to have great importance the application for accelerating wolfberry fruit extract in health care of food product.
Summary of the invention
It is low in order to provide extraction efficiency in edible lycium ruthenicum extractive of general flavone and solution Extracting Technique of Flavonoids, it is organic
The big problem of solvent consumption, the present invention provides a kind of extracting methods of the lycium ruthenicum general flavone of green high-efficient.
In order to achieve the above-mentioned object of the invention, the invention provides the following technical scheme:
1. a kind of extracting method of lycium ruthenicum general flavone, which comprises the following steps:
(1) after drying lycium ruthenicum in a vacuum drying oven, after crushed 40 meshes, the powder after sieving is recycled into pulverizer
After crushing 5-10min, lycium ruthenicum micro mist is obtained after sieving with 100 mesh sieve；
(2) after lycium ruthenicum micro mist being handled 10-15 min under microwave, with ionic liquid-salt double-aqueous phase system by 1:15-20's
Mass volume ratio after mixing, filters to get filtrate after ultrasonic extraction 30-60min under the conditions of 40-60 DEG C, and residue uses double water again
After mutually extracting 2-3 times, filtrate is stood into split-phase after merging filtrate after mixing, upper phase is collected after crossing separatory funnel, by upper phase
Cross ultrafiltration membrane remove big molecular impurity after filtrate decompression is concentrated after lycium ruthenicum general flavone extracting solution；
(3) general flavone extracting solution is loaded in chromatographic column, then successively utilizes water, 25% alcohol-water mixed solution, acetic acid second
Ester-alcohol-water mixed solution, 85% ethyl acetate-ethanol mixed solution eluted after, collect ethyl acetate, alcohol and water
Eluent when mixed solution elutes；
(4) the eluent evaporated under reduced pressure of collection is placed in super critical fluid apparatus, after purification by supercritical fluid extraction, system
It is standby to obtain lycium ruthenicum general flavone.
Preferably, the partial size of lycium ruthenicum micro mist as described in step (1) is not more than 5 μm.
Preferably, ionic liquid described in step (2)-salt double-aqueous phase system the preparation method comprises the following steps: by ionic liquid with
After salting liquid is by the volume ratio mixing of 1:3-4, after ultrasonic 305min, stands and both obtain ionic liquid-salt double-aqueous phase system；
Preferably, ionic liquid described in ionic liquid-salt double-aqueous phase system preparation method is hexafluorophosphoric acid 1- butyl -3- first
Base imidazoles ([C4mim] [PF6]), hexafluorophosphoric acid 1- hexyl -3- methylimidazole ([C6mim] [PF6]) and hexafluorophosphoric acid 1- octyl -
One of 3- methylimidazole ([C8mim] [PF6])；The salt is NaCl, KH2PO4、NaH2PO4One of；Described
The concentration of salting liquid is 0.2-0.35g/mL.
Preferably, the molecular cut off of ultrafiltration membrane described in step (2) is 10000；It is black after the reduced pressure
Total flavonoids of lycium barbarum L extracting solution density is 1.15-1.25g/mL.
Preferably, the filler in chromatographic column described in step (3) is AB-8 type macroporous absorbent resin；The acetic acid second
Ethyl acetate in ester-alcohol-water mixed solution, ethyl alcohol, water volume ratio be 35:50:15.
Preferably, supercritical fluid extraction condition described in step (4) are as follows: extracting pressure 15-20Mpa, CO2Stream
Speed is 15-18L/h, extraction time 25-30min.
Preferably, lycium ruthenicum general flavone content described in step (4) is not less than 90%.
Compared with prior art, a kind of extracting method of lycium ruthenicum general flavone of the present invention has below beneficial to effect
Fruit: United microwave processing destroys plant cell wall in lycium ruthenicum after the present invention prepares micro mist using ultramicro grinding, so that general flavone
Compound energy Rapid Leaching improves extraction efficiency；Joint ultrafiltration of the present invention, column chromatography and supercritical fluid extraction are to lycium ruthenicum
General flavone is purified, so that the lycium ruthenicum general flavone purity is high of preparation, antibacterial action are obvious；Use ionic liquid as green
Solvent extracts the general flavone in plant, the significant consumption for reducing organic solvent.Through lycium ruthenicum prepared by the present invention
General flavone can be directly as the functional additive of health care of food product.
Specific embodiment
[embodiment 1]
A kind of extracting method of lycium ruthenicum general flavone, which comprises the following steps:
(1) after drying lycium ruthenicum in a vacuum drying oven, after crushed 40 meshes, the powder after sieving is recycled into pulverizer
After crushing 8min, lycium ruthenicum micro mist is obtained after sieving with 100 mesh sieve；
(2) after lycium ruthenicum micro mist being handled 12 min under microwave, with hexafluorophosphoric acid 1- butyl -3- methylimidazole ([C4mim]
[PF6]) -0.3g/mL NaCl double-aqueous phase system by 1:12 mass volume ratio after mixing, at 45 DEG C, under the conditions of 200W surpass
Sound filters to get filtrate after extracting 45min, after residue is extracted 2-3 times with aqueous two-phase again, after merging filtrate after mixing by filtrate
Split-phase is stood, upper phase is collected after crossing separatory funnel, filters out macromolecular after upper phase is crossed the ultrafiltration membrane that molecular cut off is 10000
It is concentrated filtrate to after impurity after density is 1.15-1.25g/mL and obtains lycium ruthenicum general flavone extracting solution；
(3) general flavone extracting solution is loaded in AB-8 type macroporous adsorption resin chromatography column, then successively utilizes water, 25% ethyl alcohol-
Water mixed solution, ethyl acetate, alcohol and water mixed solution, 85% ethyl acetate-ethanol mixed solution eluted after, collect
Eluent when ethyl acetate, alcohol and water mixed solution elutes；
(4) liquid pressure-reducing of collection is evaporated and is placed in super critical fluid apparatus, be 15Mpa, CO in extracting pressure2Flow velocity
For 15L/h, extraction time be 25min under the conditions of after purification, lycium ruthenicum general flavone is prepared.
[embodiment 2]
A kind of extracting method of lycium ruthenicum general flavone, which comprises the following steps:
(1) after drying lycium ruthenicum in a vacuum drying oven, after crushed 40 meshes, the powder after sieving is recycled into pulverizer
After crushing 8min, lycium ruthenicum micro mist is obtained after sieving with 100 mesh sieve；
(2) after lycium ruthenicum micro mist being handled 12 min under microwave, with hexafluorophosphoric acid 1- hexyl -3- methylimidazole ([C6mim]
[PF6])-0.3g/mL Na2HPO4Double-aqueous phase system presses the mass volume ratio of 1:17 after mixing, at 45 DEG C, under the conditions of 200W
It filters to get filtrate after ultrasonic extraction 45min, after residue is extracted 2-3 times with aqueous two-phase again, is uniformly mixed filtrate after merging filtrate
After stand split-phase, collect upper phase after crossing separatory funnel, upper phase crossed after the ultrafiltration membrane that molecular cut off is 10000 and filters out big point
It is concentrated filtrate to after sub- impurity after density is 1.15-1.25g/mL and obtains lycium ruthenicum general flavone extracting solution；
(3) general flavone extracting solution is loaded in AB-8 type macroporous adsorption resin chromatography column, then successively utilizes water, 25% ethyl alcohol-
Water mixed solution, ethyl acetate, alcohol and water mixed solution, 85% ethyl acetate-ethanol mixed solution eluted after, collect
Eluent when ethyl acetate, alcohol and water mixed solution elutes；
(4) liquid pressure-reducing of collection is evaporated and is placed in super critical fluid apparatus, be 17Mpa, CO in extracting pressure2Flow velocity
For 15L/h, extraction time be 30min under the conditions of after purification, lycium ruthenicum general flavone is prepared.
[embodiment 3]
A kind of extracting method of lycium ruthenicum general flavone, which comprises the following steps:
(1) after drying lycium ruthenicum in a vacuum drying oven, after crushed 40 meshes, the powder after sieving is recycled into pulverizer
After crushing 10min, lycium ruthenicum micro mist is obtained after sieving with 100 mesh sieve；
(2) after lycium ruthenicum micro mist being handled 12 min under microwave, with hexafluorophosphoric acid 1- octyl -3- methylimidazole ([C8mim]
[PF6])-0.25g/mL K2HPO4Double-aqueous phase system presses the mass volume ratio of 1:18 after mixing, at 45 DEG C, under the conditions of 200W
It filters to get filtrate after ultrasonic extraction 45min, after residue is extracted 2-3 times with aqueous two-phase again, is uniformly mixed filtrate after merging filtrate
After stand split-phase, collect upper phase after crossing separatory funnel, upper phase crossed after the ultrafiltration membrane that molecular cut off is 10000 and filters out big point
It is concentrated filtrate to after sub- impurity after density is 1.15-1.25g/mL and obtains lycium ruthenicum general flavone extracting solution；
(3) general flavone extracting solution is loaded in AB-8 type macroporous adsorption resin chromatography column, then successively utilizes water, 25% ethyl alcohol-
Water mixed solution, ethyl acetate, alcohol and water mixed solution, 85% ethyl acetate-ethanol mixed solution eluted after, collect
Eluent when ethyl acetate, alcohol and water mixed solution elutes；
(4) liquid pressure-reducing of collection is evaporated and is placed in super critical fluid apparatus, be 15Mpa, CO in extracting pressure2Flow velocity
For 17L/h, extraction time be 25min under the conditions of after purification, lycium ruthenicum general flavone is prepared.
The measurement of lycium ruthenicum general flavone content:
Specification Curve of Increasing: taking 6 10ml volumetric flasks, is separately added into 1.0ml, 2.0ml, 3.0ml, 4.0ml and 5.0ml rutin mark
Liquid (0.2mg/mL) adds 80% ethyl alcohol to add 5%NaN0 to 5mL2Solution 0.5ml shakes up and places 6min, adds 1O%Al
(N03)3Solution 0.5ml shakes up, then places 6min, and 4%NaOH solution 4ml is added, shakes up, and places 10min, with UV, visible light point
Light photometer measures its absorbance at 510nm, lycium ruthenicum general flavone extracting solution is not added as blank control.With rutin standard specimen
Concentration (mg/m1) is abscissa X, and absorbance value A is ordinate Y, draws standard curve, obtains regression equation Y=1.312X-
0.101,R=0.999。
The measurement of lycium ruthenicum general flavone content:
Lycium ruthenicum general flavone made from embodiment 1-3 is configured to the solution of 0.2mg/mL, takes 1 mL solution in 10mL capacity
In bottle, 80% ethyl alcohol is added to add 5%NaN0 to 5mL2Solution 0.5ml shakes up and places 6min, adds 1O%Al (N03)3It is molten
Liquid 0.5ml shakes up, then places 6min, and 4%NaOH solution 4ml is added, shakes up, puts 10min, existed with ultraviolet-uisible spectrophotometer
Its absorbance is measured at 510nm, and measures its concentration by standard curve and calculates its content, is calculated general flavone yield, is as a result seen
Table 1.90% or more is up to through lycium ruthenicum general flavone purity prepared by the present invention as can be seen from Table 1.
The content and its yield of gained lycium ruthenicum general flavone in 1. embodiment 1-3 of table
| Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| Lycium ruthenicum general flavone content (&) | 91.5 | 92.2 | 91.8 |
| Yield (%) | 2.7 | 2.8 | 2.6 |
The test of lycium ruthenicum general flavone bacteriostatic activity:
On superclean bench, sterilized Oxford cup is vertically placed in sterile petri dish with aseptic nipper, each culture
Ware places 3.By prepared each bacterium culture medium moist heat sterilization after it is cooled to 50 DEG C~55 DEG C, in sterile behaviour
Under work, the ratio for being 1: 100 according to the ratio between bacteria suspension and culture volume, the bacteria suspension for drawing corrected concentrations adds respectively
Enter in each corresponding culture medium, after concussion mixes, is poured slowly into the sterile petri dish for having placed Oxford cup.It is solidified to culture medium
Afterwards, Oxford cup is extracted with sterilized tweezers.Then, the molten of the general flavone configuration of the embodiment 1-3 of 100 μ L is drawn respectively
Liquid (0.5mg/mL) uses culture medium added with 20 % ethyl alcohol as blank control in 3 holes.Finally culture medium is put into
Culture in constant incubator (35 DEG C~37 DEG C of bacterium, 28 DEG C of mould, saccharomycete), and a period of time (bacterium 24 h, it is mould
Bacterium, saccharomycete 72 h) observe the growing state of thallus afterwards and measure its antibacterial circle diameter, and acquired results are shown in Table 2.It can by table 2
Know, the ethanol extract of lycium ruthenicum general flavone has obvious inhibiting effect to Escherichia coli, staphylococcus aureus.
The antibacterial circle diameter of 2. lycium ruthenicum general flavone extracting solution of table
| For trying bacterium | Embodiment 1 | Embodiment 2 | Embodiment 3 | Blank |
| Escherichia coli | 1.4cm | 1.3cm | 1.3cm | - |
| Staphylococcus aureus | 1.02cm | 1.03cm | 1.01cm | - |
| Mould | 1.18cm | 1.22cm | 1.25cm | - |
Note: 3 kinds of test bacterium bacteria suspension initial concentrations are the cfu/mL of 106 cfu/mL~107；Indicate that no inhibition zone produces
It is raw.
For the ordinary skill in the art, specific embodiment is only exemplarily described the present invention,
Obviously the present invention specific implementation is not subject to the restrictions described above, as long as use the inventive concept and technical scheme of the present invention into
The improvement of capable various unsubstantialities, or not improved the conception and technical scheme of the invention are directly applied to other occasions
, it is within the scope of the present invention.
Claims (8)
1. a kind of extracting method of lycium ruthenicum general flavone, which comprises the following steps:
(1) after drying lycium ruthenicum in a vacuum drying oven, after crushed 40 meshes, the powder after sieving is recycled into pulverizer
After crushing 5-10min, lycium ruthenicum micro mist is obtained after sieving with 100 mesh sieve；
(2) after lycium ruthenicum micro mist being handled 10-15 min under microwave, with ionic liquid-salt double-aqueous phase system by 1:15-20's
Mass volume ratio after mixing, filters to get filtrate after ultrasonic extraction 30-60min under the conditions of 40-60 DEG C, and residue uses double water again
After mutually extracting 2-3 times, filtrate is stood into split-phase after merging filtrate after mixing, upper phase is collected after crossing separatory funnel, by upper phase
Cross ultrafiltration membrane remove big molecular impurity after filtrate decompression is concentrated after lycium ruthenicum general flavone extracting solution；
(3) general flavone extracting solution is loaded in chromatographic column, then successively utilizes water, 25% alcohol-water mixed solution, acetic acid second
Ester-alcohol-water mixed solution, 85% ethyl acetate-ethanol mixed solution eluted after, collect ethyl acetate, alcohol and water
Eluent when mixed solution elutes；
(4) the eluent evaporated under reduced pressure of collection is placed in super critical fluid apparatus, after purification by supercritical fluid extraction, system
It is standby to obtain lycium ruthenicum general flavone.
2. a kind of extracting method of lycium ruthenicum general flavone according to claim 1, it is characterised in that as described in step (1)
The partial size of lycium ruthenicum micro mist is not more than 5 μm.
3. a kind of extracting method of lycium ruthenicum general flavone according to claim 1, it is characterised in that described in step (2)
After ionic liquid-salt double-aqueous phase system is the preparation method comprises the following steps: ionic liquid is mixed with salting liquid by the volume ratio of 1:3-4, surpass
After sound 305min, stands and both obtain ionic liquid-salt double-aqueous phase system.
4. a kind of extracting method of lycium ruthenicum general flavone according to claim 3, it is characterised in that the ionic liquid
For hexafluorophosphoric acid 1- butyl -3- methylimidazole ([C4mim] [PF6]), hexafluorophosphoric acid 1- hexyl -3- methylimidazole ([C6mim]
One of [PF6]) and hexafluorophosphoric acid 1- octyl -3- methylimidazole ([C8mim] [PF6])；The salt is NaCl, KH2PO4、
NaH2PO4One of；The concentration of the salting liquid is 0.2-0.35g/mL.
5. a kind of extracting method of lycium ruthenicum general flavone according to claim 1, it is characterised in that described in step (2)
The molecular cut off of ultrafiltration membrane is 10000；Lycium ruthenicum general flavone extracting solution density after the reduced pressure is 1.15-
1.25g/mL。
6. a kind of extracting method of lycium ruthenicum general flavone according to claim 1, it is characterised in that described in step (3)
Filler in chromatographic column is AB-8 type macroporous absorbent resin；Ethyl acetate in the ethyl acetate, alcohol and water mixed solution,
Ethyl alcohol, water volume ratio be 35:50:15.
7. a kind of extracting method of lycium ruthenicum general flavone according to claim 1, it is characterised in that described in step (4)
Supercritical fluid extraction condition are as follows: extracting pressure 15-20Mpa, CO2Flow velocity is 15-18L/h, extraction time 25-
30min。
8. a kind of extracting method of lycium ruthenicum general flavone according to claim 1, it is characterised in that described in step (4)
Lycium ruthenicum general flavone content is not less than 90%.
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| CN109908216A (en) * | 2019-04-30 | 2019-06-21 | 湖南中医药大学 | A kind of extracting method of compound object and its application in treatment agitated depression |
| CN110934922A (en) * | 2019-12-12 | 2020-03-31 | 安徽工业大学 | Total flavone in lycium ruthenicum, extraction method and application |
| CN111840349A (en) * | 2020-07-01 | 2020-10-30 | 江汉大学 | Method for simultaneously extracting polysaccharide and flavonoid components from folium artemisiae argyi |
| CN112587450A (en) * | 2020-12-18 | 2021-04-02 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | Lycium ruthenicum extract with ultraviolet resistance and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109908216A (en) * | 2019-04-30 | 2019-06-21 | 湖南中医药大学 | A kind of extracting method of compound object and its application in treatment agitated depression |
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| CN111840349B (en) * | 2020-07-01 | 2022-08-30 | 江汉大学 | Method for simultaneously extracting polysaccharide and flavonoid components from folium artemisiae argyi |
| CN112587450A (en) * | 2020-12-18 | 2021-04-02 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | Lycium ruthenicum extract with ultraviolet resistance and preparation method thereof |
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Application publication date: 20181218 |