CN106668234A - Process for extracting and purifying total flavone from rose flower - Google Patents
Process for extracting and purifying total flavone from rose flower Download PDFInfo
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- CN106668234A CN106668234A CN201611255018.1A CN201611255018A CN106668234A CN 106668234 A CN106668234 A CN 106668234A CN 201611255018 A CN201611255018 A CN 201611255018A CN 106668234 A CN106668234 A CN 106668234A
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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- 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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- 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 invention provides a process for extracting and purifying total flavone from rose flower. The total flavone provided by the invention is obtained by extracting dry petals of plant rose flower by using a solvent, and purifying by using macroporous resin, polyamide and C18; the total flavone uses flavonoid compounds as main components. The invention provides a simple, convenient and easily-implemented method for determining the content of the total flavone in the rose flower, and optimizes the process for extracting and purifying the total flavone from the rose flower; the content of the final total flavone from the rose flower is obviously increased, so that the process is wide in application prospect.
Description
Technical field
The present invention relates to the technical field of total flavones is extracted from Chinese medicinal plant, and in particular to a kind of Flos Rosae Rugosae total flavones are carried
Take purifying process.
Background technology
Flos Rosae Rugosae is rosaceous plant Flos Rosae Rugosae（RosarugosaThunb）Dry flower, which is sweet in flavor and warm in property, Return liver, spleen
Jing, the effect of with promoting QI circulation for relieving depression, regulating blood flow to alleviate pains.Flos Rosae Rugosae contains volatile oil, aminoacid, polysaccharide and various flavonoids
Thing, flavones ingredient therein have various pharmacologically actives such as anticancer, immunomodulating, blood sugar lowering, antioxidation.But it is current, from
The process route that total flavones are provided in Flos Rosae Rugosae is immature, is mainly shown as that the effective ingredient total amount of extraction is not high（That is purity
It is low）, process route is not reasonable or industrialized condition does not possess.Such as Publication No. CN102796066A, a kind of entitled Huang
Ketone compounds and preparation method and application, with Flos Rosae Rugosae as raw material, the extraction of Jing extractum, silica gel column chromatography, petroleum ether~second
Acetoacetic ester column chromatography, high pressure liquid chromatography separating step, specially：Flos Rosae Rugosae is taken, is crushed, with 90~99% ethanol ultrasonic extraction
United extraction liquid, filtration, concentrating under reduced pressure is into extractum；Extractum carries out silica gel column chromatography with silica gel dry column-packing；With volume proportion it is
20:1~1:1 chloroform~methanol solution carries out gradient elution, merges identical part；9:1 part volume proportion is 9:1~
0:1 petroleum ether~ethyl acetate solution carries out gradient elution, collects each several part eluent and concentrates；7:Further use 3 parts
High pressure liquid chromatography is isolated and purified and obtains final product described flavone compound.It is most to there is following problems flavone compound in the method
With phenolic hydroxyl group, alkaline water or dilute alcohol are soluble in（50%~70%）In, the method is extracted using 90~99% ethanol, flavonoid thing
Dissolubility is relatively low wherein for matter, and silica gel can form dead absorption to part flavone, cause product to lose so that flavone yield compared with
It is low.
And for example Publication No. CN104083476A, a kind of entitled Flos Rosae Rugosae extract in antidepressant drug is prepared should
With and Publication No. CN103417679A, in a kind of entitled Flos Rosae Rugosae extract anti-cerebral ischemia material method and its application, be
Flos Rosae Rugosae is dried and crushed into into coarse powder, with petroleum ether reflux extraction defat, petroleum ether liquid is discarded, the Flos Rosae Rugosae medicinal residues after defat are waved
Dry petroleum ether, soaks reflux, extract, with ethanol, and to without alcohol taste, concentration, concentrated solution add distilled water to disperse to decompression filtrate recycling ethanol；
Type macroporous adsorbent resin in AB~8 on dispersion liquid, with distillation water elution, discards aqueous；After with ethanol elution, eluent is discarded；Again
With ethanol elution, eluent is collected, decompression recycling ethanol, drying and crushing are obtained final product, its total flavones mass content is more than 63%.Such as
Publication No. CN103006899A, a kind of entitled extraction and purification process of Fructus Lycii seed total flavonoid, detection method and its application, carries
Taking technique is Fructus Lycii dries pulverizing, adds ethanol, extracts merging filtrate, concentrates, extracts, obtaining Fructus Lycii flavone crude extract；Carry
Take purifying process：Using macroporous resin D101, purification is carried out to crude extract, using polyamide to further purification of flavone etc.,
Its flavone purity only has 42.8%.
The content of the invention
Present invention seek to address that the purity that Flos Rosae Rugosae extraction and purification process for total flavonoids is present in prior art and yield is low asks
Topic, there is provided a kind of method is reasonable, stable, with short production cycle, the high Flos Rosae Rugosae of the total flavones purity for being adapted to industrialized production and obtaining
The extraction and purification process of flower total flavones.
To solve above-mentioned technical problem, the present invention is achieved through the following technical solutions：
Flos Rosae Rugosae extraction and purification process for total flavonoids, comprises the steps：
A, take Flos Rosae Rugosae and be dried medical material, after crushing, add alcohol reflux, extracting solution is filtered, standby；
B, the extracting solution for obtaining step A remove ethanol, obtain once concentration liquid, and once concentration liquid is added to AB~8 type macropore
In resin, Jing water, 5~10% ethanol are washed till clarification of water successively, and ethanol color is shallower, finally with 80~85% ethanol elutions, eluting
Liquid removes ethanol, obtains secondary concentration liquid；
C, the secondary concentration liquid obtained in step B is poured in the polyamide column for installing, water is washed till with water, 5~10% ethanol successively
Clarification, ethanol color are shallower, and finally with 70~75% ethanol elutions, eluent is concentrated to dryness, and obtains Flos Rosae Rugosae total flavones crude product；
D, the Flos Rosae Rugosae total flavones crude product methanol obtained in step C is dissolved, sample, dry sample are mixed with polyamide, then
Sample dry method is loaded to into column chromatography polyamide for filler, dichloromethane and methanol are, on the pillar of solvent wet method dress post, to adopt
Dichloromethane：Methanol is 10：1~1：1 ratio carries out gradient elution, collects the eluent containing total flavones according to TLC points plate,
Eluent is concentrated to dryness to obtain purification Flos Rosae Rugosae total flavones；
Filter after E, the purification Flos Rosae Rugosae total flavones methanol for obtaining step D dissolving, filtrate to entering in C18 posts, with 20%~
70% methanol solution is solvent, in 254nm wavelength Gradient eluting, collects elution fractions containing flavone, and recovery methanol is simultaneously dense
Contracting, concentrated solution is enriched with C18 posts, is then parsed with methanol, collects methanol desorbed solution, and desorbed solution removes methanol and is concentrated to dryness
Obtain Flos Rosae Rugosae total flavones.
Further, the technique also includes that the Flos Rosae Rugosae total flavones for obtaining step E are carried out again according to the operation of step D
Process, i.e., the dissolving of step E is obtained Flos Rosae Rugosae total flavones methanol, sample, dry sample are mixed with polyamide, then sample is done
Method is loaded to column chromatography polyamide for filler, dichloromethane and methanol on the pillar of solvent wet method dress post, using dichloromethane：
Methanol is 10：1~1：1 ratio carries out gradient elution, collects the eluent containing total flavones, eluent concentration according to TLC points plate
It is extremely dry.
Preferably, in step, 5~15 times for Flos Rosae Rugosae medical material weight of the addition of the ethanol, the ethanol
Volumetric concentration is 70~75%, reflux, extract, 2~3 times, 1.5~2 hours every time.
In step D, the column chromatography polyamide consumption is 80~120 times of Flos Rosae Rugosae total flavones crude product gross weight.
In step D, during the wet method dress post, the volume ratio of dichloromethane and methanol is 10:1.
In step D, using dichloromethane：Methanol is 10：1~1：It is 10 that 1 ratio carries out the process of gradient elution：1 stream
Dynamic phase eluting 15L, 8：1 mobile phase eluting 10L, 6：1 mobile phase eluting 10L, 3：1 mobile phase eluting 10L, 1：1 mobile phase eluting
15L, TLC point plate collects the eluent part of flavone reaction.
In step E, the process of 20%~70% methanol solution gradient elution is to be prepared using binary pump gradient, often
2min Fractional Collectionses, are detected by TLC points plate, collect the eluent of flavone reaction.
In step E, the parsing methanol concentration is 90%.
The Flos Rosae Rugosae total flavones that extraction purification is obtained can be detected as steps described below：
A instruments and reagent
Instrument：Ultraviolet~visible spectrophotometric instrument
Reagent：Control substance of Rutin, methanol, sodium nitrite, aluminum nitrate, sodium hydroxide；
It is prepared by B solution
It is prepared by A, reference substance stock solution：Precision weighs control substance of Rutin 50mg, is placed in 25ml volumetric flasks, plus appropriate methanol, puts
In water-bath, slight fever dissolving, lets cool, plus methanol is to scale, shakes up.Precision measures 10ml, puts in 100ml volumetric flasks, adds water to quarter
Degree, shakes up, obtains final product（0.2mg containing rutin in per 1ml）；
The drafting of B, standard curve：Precision measures reference substance solution 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, is respectively placed in 25ml appearances
In measuring bottle, 6.0ml, plus 5% sodium nitrite solution 1ml are respectively added water to, mixed, placed 6 minutes, hydro-oxidation sodium test solution 10ml, then
Scale is added water to, is shaken up, placed 15 minutes, with corresponding reagent as blank, using ultraviolet~visible spectrophotometric instrument, in 500nm
Mensuration absorbance at wavelength, with absorbance as vertical coordinate, concentration is abscissa, draws standard curve, obtains regression equation：A=
8.1046C+0.0113, correlation coefficient γ=0.9985.
It is prepared by C, need testing solution：Precision weighs sample 40mg, puts in 10ml measuring bottles, adds methanol appropriate, in water-bath
Slight fever dissolve, let cool, plus methanol be to scale, after be placed in 1000ml measuring bottles, add water to graduation mark, shake up, obtain final product；
It is prepared by D, blank solution：Using corresponding solution as blank；
C, sample determination
It is accurate to draw need testing solution 3ml, it is placed in 25ml volumetric flasks, the method under sighting target directrix curve preparation is added water to
6.0ml, plus 5% sodium nitrite solution 1ml, mix, and place 6 minutes, and hydro-oxidation sodium test solution 10ml is added water to scale, shaken
It is even, place 15 minutes, with corresponding reagent as blank, using ultraviolet~visible spectrophotometric instrument, extinction is determined at 500nm wavelength
Degree, general flavone content are calculated with rutin；
D results are calculated
From the content of total flavones in regression equation calculation need testing solution.The inventive method extraction purification is detected using the method
Flos Rosae Rugosae total flavones, general flavone content are calculated more than or equal to 90% with rutin.
The invention has the advantages that：
The present invention by the extracting solution that Flos Rosae Rugosae is dried medical material sequentially pass through AB-8 macroporous resins, polyamide column, column chromatography polyamide,
C18 posts this specific purification procedures, so as to prepare the Flos Rosae Rugosae total flavones that content is up to more than 90%, are herb resource
Make full use of and improve the foundation that its using value provides science.
2nd, the present invention is reasonable in design, rational technology, stable, feasible, is extracted using the inventive method, total in purification Flos Rosae Rugosae
Flavone, separating rate are fast, with short production cycle, are adapted to industrialized production, have good application prospect.
2nd, the present invention is always added to AB-8 macroporous resins after extracting solution is concentrated, and is washed with water and low-concentration ethanol successively,
Water colo(u)r and impurity such as saccharide are eliminated, the pigment and impurity of big polarity is removed, and all without by under object eluting
Come；Facilitate follow-up polyamide to separate with C18, reduce the pollution to filler；AB-8 type macroporous resins are a kind of adsorption ranges compared with
Wide filler, can adsorb most pigment and impurity.
3rd, the present invention will further use polyamide column subprocessing after AB-8 type macroporous resins, and polyamide filler is to flavone
Class material has more single-minded separating effect, isolates and purifies efficiency to improve, using water and low is first used before high concentration ethanol eluting
Concentration ethanol is washed, first depigmentation and impurity, only just need to can make the Flavonoid substances part under eluting at high proportion through eluting successively
It is purer, next step positive eluting is more convenient, while avoiding the materials such as impurity, polysaccharide from polluting to later separation post and block.
4th, sample is mixed sample using polyamide by the present invention can make total flavones crude product uniformly disperse to be attracted to polyamide
On, when adding mobile phase eluting, sample is uniformly dissolved so that during gradient elution impurity and product slates evenly, concentrate, reach
Efficiently separate；Drying after mixing sample makes the methanol in sample volatilize completely, allows polyamide to mix in sample without methanol, it is to avoid to wash in gradient
Change mobile phase polarity during de- initial stage low polar organic solvent eluting, and it is bad to cause impurity to separate with product.
5th, the rose extract through AB-8 macroporous resins, polyamide column, the process of column chromatography polyamide is entered one by the present invention
Step is processed with C18 posts, can be oriented and be obtained most of Flavonoid substances and make flavones content improve a lot, and disengaging time is shorter,
And one or more flavonoid substances therein can be specified, be conducive to which further to develop.
6th, the Flos Rosae Rugosae after AB-8 macroporous resins, polyamide column, column chromatography polyamide, the process of C18 posts is carried by the present invention
Thing is taken again through the process of column chromatography polyamide, not only can further improve the purity of total flavones, flavones content is brought up to into 90%
More than；And the yield of total flavones can be improved, operate easier to be quick.
Specific embodiment
Embodiment 1
Flos Rosae Rugosae extraction and purification process for total flavonoids, comprises the steps：
A, take Flos Rosae Rugosae and be dried medical material, after crushing, add Flos Rosae Rugosae to be dried 70% alcohol reflux 3 times of 5 times of medical material weight,
2 hours every time, extracting solution was filtered, merging filtrate；
B, the extracting solution for obtaining step A remove ethanol, obtain once concentration liquid, and once concentration liquid is added to AB~8 type macropore
In resin, Jing water, 10% ethanol are washed till clarification of water successively, and ethanol color is shallower, finally use 80% ethanol elution, eluent to remove second
Alcohol, obtains secondary concentration liquid；
C, the secondary concentration liquid obtained in step B is poured in the polyamide column for installing, successively with water, that 10% ethanol is washed till water is clear
Clearly, ethanol color is shallower, finally uses 70% ethanol elution, eluent to be concentrated to dryness, and obtains Flos Rosae Rugosae total flavones crude product；
D, the Flos Rosae Rugosae total flavones crude product methanol obtained in step C is dissolved, the addition of methanol is thick with Flos Rosae Rugosae total flavones
Product all preferably, mix sample, dry sample with polyamide after dissolving, sample dry method be loaded to column chromatography polyamide then by dissolving
For filler, dichloromethane and methanol on the pillar of solvent wet method dress post, using dichloromethane：Methanol is 10：1~1：1 ratio
Example carry out gradient elution, according to TLC points plate collect containing total flavones eluent, eluent be concentrated to dryness purification Flos Rosae Rugosae is always yellow
Ketone；
Filter after E, the purification Flos Rosae Rugosae total flavones methanol for obtaining step D dissolving, filtrate to entering in C18 posts, with 20%~
70% methanol solution is solvent, in 254nm wavelength Gradient eluting, collects elution fractions containing flavone, and recovery methanol is simultaneously dense
Contracting, concentrated solution is enriched with C18 posts, is then parsed with methanol, collects methanol desorbed solution, and desorbed solution removes methanol and is concentrated to dryness
Obtain Flos Rosae Rugosae total flavones.
Determine and calculate, general flavone content calculates total flavones with rutin in the present embodiment Flos Rosae Rugosae extractive of general flavone
Content is 90.10%.
Embodiment 2
The present embodiment with the difference of embodiment 1 is：The present embodiment also includes the Flos Rosae Rugosae total flavones for obtaining step E again
Processed according to step D process, i.e., dissolved with methanol, sample, dry sample are mixed with polyamide, then by sample dry method loading
Be filler to column chromatography polyamide, dichloromethane and methanol on the pillar of solvent wet method dress post, using dichloromethane：Methanol is
10：1~1：1 ratio carries out gradient elution, collects the eluent containing total flavones according to TLC points plate, and eluent is concentrated to dryness.Its
In, in the step, the column chromatography polyamide consumption is 120 times of Flos Rosae Rugosae total flavones crude product gross weight.Use during the wet method dress post
The volume ratio of dichloromethane and methanol is 10:1.
Determine and calculate, general flavone content calculates total flavones with rutin in the present embodiment Flos Rosae Rugosae extractive of general flavone
Content is 91.16%
Embodiment 3
Flos Rosae Rugosae extraction and purification process for total flavonoids, comprises the steps：
A, take Flos Rosae Rugosae and be dried medical material, after crushing, add Flos Rosae Rugosae to be dried 75% alcohol reflux 3 times of 10 times of medical material weight,
2 hours every time, extracting solution was filtered, merging filtrate；
B, the extracting solution for obtaining step A remove ethanol, obtain once concentration liquid, and once concentration liquid is added to AB~8 type macropore
In resin, Jing water, 5% ethanol are washed till clarification of water successively, and ethanol color is shallower, finally use 85% ethanol elution, eluent to remove second
Alcohol, obtains secondary concentration liquid；
C, the secondary concentration liquid obtained in step B is poured in the polyamide column for installing, successively with water, that 5% ethanol is washed till water is clear
Clearly, ethanol color is shallower, finally uses 75% ethanol elution, eluent to be concentrated to dryness, and obtains Flos Rosae Rugosae total flavones crude product；
D, the Flos Rosae Rugosae total flavones crude product methanol obtained in step C is dissolved, sample, dry sample are mixed with polyamide, then
Sample dry method is loaded to into column chromatography polyamide for filler, dichloromethane and methanol are, on the pillar of solvent wet method dress post, to adopt
Dichloromethane：Methanol is 10：1~1：1 ratio carries out gradient elution, collects the eluent containing total flavones according to TLC points plate, washes
De- liquid is concentrated to dryness to obtain purification Flos Rosae Rugosae total flavones；
Filter after E, the purification Flos Rosae Rugosae total flavones methanol for obtaining step D dissolving, filtrate to entering in C18 posts, with 20%~
70% methanol solution is solvent, in 254nm wavelength Gradient eluting, collects elution fractions containing flavone, and recovery methanol is simultaneously dense
Contracting, concentrated solution is enriched with C18 posts, is then parsed with methanol, collects methanol desorbed solution, and desorbed solution removes methanol and is concentrated to dryness
Obtain Flos Rosae Rugosae total flavones.
F, the Flos Rosae Rugosae total flavones methanol for obtaining step E dissolving, mix sample, dry sample with polyamide, then will
Sample dry method is loaded to column chromatography polyamide for filler, and dichloromethane and methanol are, on the pillar of solvent wet method dress post, to adopt two
Chloromethanes：Methanol elution gradient, according to TLC points plate collect containing total flavones eluent, eluent be concentrated to dryness Flos Rosae Rugosae is total
Flavone finished product.
In above-mentioned steps D and F, the column chromatography polyamide consumption is 80 times of Flos Rosae Rugosae total flavones crude product gross weight.It is described wet
During method dress post, the volume ratio of dichloromethane and methanol is 10:1.
In step E, the parsing methanol concentration is 90%.
Determine and calculate, general flavone content calculates total flavones with rutin in the present embodiment Flos Rosae Rugosae extractive of general flavone
Content is 93.15%.
Embodiment 4
The extraction and purification process of Flos Rosae Rugosae total flavones, comprises the steps：A, take Flos Rosae Rugosae and be dried medical material, after crushing, add rose
Rare to spend 70% alcohol reflux 3 times for being dried 15 times of medical material weight, 2 hours every time, extracting solution was filtered, merging filtrate；
B, the extracting solution for obtaining step A remove ethanol, obtain once concentration liquid, and once concentration liquid is added to AB~8 type macropore
In resin, Jing water, 8% ethanol are washed till clarification of water successively, and ethanol color is shallower, finally use 80% ethanol elution, eluent to remove second
Alcohol, obtains secondary concentration liquid；
C, the secondary concentration liquid obtained in step B is poured in the polyamide column for installing, successively with water, that 8% ethanol is washed till water is clear
Clearly, ethanol color is shallower, finally uses 72% ethanol elution, eluent to be concentrated to dryness, and obtains Flos Rosae Rugosae total flavones crude product；
D, the Flos Rosae Rugosae total flavones crude product methanol obtained in step C is dissolved, sample, dry sample are mixed with polyamide, then
Sample dry method is loaded to into column chromatography polyamide for filler, dichloromethane and methanol are, on the pillar of solvent wet method dress post, to adopt
Dichloromethane：Methanol is 10：1~1：1 ratio carries out gradient elution, collects the eluent containing total flavones according to TLC points plate, washes
De- liquid is concentrated to dryness to obtain purification Flos Rosae Rugosae total flavones；
Filter after E, the purification Flos Rosae Rugosae total flavones methanol for obtaining step D dissolving, filtrate to entering in C18 posts, with 20%~
70% methanol solution is solvent, in 254nm wavelength Gradient eluting, collects elution fractions containing flavone, and recovery methanol is simultaneously dense
Contracting, concentrated solution is enriched with C18 posts, is then parsed with methanol, collects methanol desorbed solution, and desorbed solution removes methanol and is concentrated to dryness
Obtain Flos Rosae Rugosae total flavones.
F, the Flos Rosae Rugosae total flavones methanol for obtaining step E dissolving, mix sample, dry sample with polyamide, then will
Sample dry method is loaded to column chromatography polyamide for filler, and dichloromethane and methanol are, on the pillar of solvent wet method dress post, to adopt two
Chloromethanes：Methanol elution gradient, according to TLC points plate collect containing total flavones eluent, eluent be concentrated to dryness Flos Rosae Rugosae is total
Flavone finished product.
In above-mentioned steps D and F, the column chromatography polyamide consumption is 100 times of Flos Rosae Rugosae total flavones crude product gross weight.It is described wet
During method dress post, the volume ratio of dichloromethane and methanol is 10:1.
In step E, the parsing methanol concentration is 90%.
Determine and calculate, general flavone content calculates total flavones with rutin in the present embodiment Flos Rosae Rugosae extractive of general flavone
Content is 91.50%.
In above-described embodiment 1-4, in step D, using dichloromethane：Methanol is 10：1~1：1 ratio carries out gradient and washes
De- process is 10：1 mobile phase eluting 15L, 8：1 mobile phase eluting 10L, 6：1 mobile phase eluting 10L, 3：1 mobile phase eluting
10L, 1：1 mobile phase eluting 15L, TLC points plate collect the eluent part of flavone reaction.In step E, 20%~70% first
The process of alcoholic solution gradient elution is to be prepared using binary pump gradient, per 2min Fractional Collectionses, is detected by TLC points plate, is collected
The eluent of flavone reaction.
Claims (9)
1. Flos Rosae Rugosae extraction and purification process for total flavonoids, it is characterised in that：The technique is comprised the following steps：
A, take Flos Rosae Rugosae and be dried medical material, after crushing, add alcohol reflux, extracting solution is filtered, standby；
B, the extracting solution for obtaining step A remove ethanol, obtain once concentration liquid, and once concentration liquid is added to AB-8 type macropore trees
In fat, Jing water, 5~10% ethanol are washed till clarification of water successively, and ethanol color is shallower, finally with 80~85% ethanol elutions, eluent
Ethanol is removed, secondary concentration liquid is obtained；
C, the secondary concentration liquid obtained in step B is poured in the polyamide column for installing, water is washed till with water, 5~10% ethanol successively
Clarification, ethanol color are shallower, and finally with 70~75% ethanol elutions, eluent is concentrated to dryness, and obtains Flos Rosae Rugosae total flavones crude product；
D, the Flos Rosae Rugosae total flavones crude product methanol obtained in step C is dissolved, sample, dry sample are mixed with polyamide, then
Sample dry method is loaded to into column chromatography polyamide for filler, dichloromethane and methanol are, on the pillar of solvent wet method dress post, to adopt
Dichloromethane：Methanol is 10：1~1：1 ratio carries out gradient elution, collects the eluent containing total flavones according to TLC points plate,
Eluent is concentrated to dryness to obtain purification Flos Rosae Rugosae total flavones；
Filter after E, the purification Flos Rosae Rugosae total flavones methanol for obtaining step D dissolving, filtrate to entering in C18 posts, with 20%~
70% methanol solution is solvent, in 254nm wavelength Gradient eluting, collects elution fractions containing flavone, and recovery methanol is simultaneously dense
Contracting, concentrated solution is enriched with C18 posts, is then parsed with methanol, collects methanol desorbed solution, and desorbed solution removes methanol and is concentrated to dryness
Obtain Flos Rosae Rugosae total flavones.
2. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 1, it is characterised in that：Also include obtaining step E
The Flos Rosae Rugosae total flavones methanol dissolving arrived, mixes sample, dry sample with polyamide, sample dry method is loaded to column chromatography then
Polyamide is filler, dichloromethane and methanol on the pillar of solvent wet method dress post, using dichloromethane：Methanol is 10：1~1：
1 ratio carries out gradient elution, collects the eluent containing flavone according to TLC points plate, and eluent is concentrated to dryness.
3. the extraction and purification process of Flos Rosae Rugosae total flavones according to claim 1, it is characterised in that：In step, it is described
The addition of ethanol is 5~15 times of Flos Rosae Rugosae medical material weight, and the volumetric concentration of the ethanol is 70~75%, reflux, extract, 2~
3 times, every time 1.5~2 hours.
4. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 1, it is characterised in that：In step D, the post
Chromatography polyamide consumption is 80~120 times of Flos Rosae Rugosae total flavones crude product gross weight.
5. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 1, it is characterised in that：In step D, it is described wet
During method dress post, the volume ratio of dichloromethane and methanol is 10:1.
6. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 1, it is characterised in that：In step D, two are adopted
Chloromethanes：Methanol is 10：1~1：It is 10 that 1 ratio carries out the process of gradient elution：1 mobile phase eluting 15L, 8：1 mobile phase is washed
De- 10L, 6：1 mobile phase eluting 10L, 3：1 mobile phase eluting 10L, 1：1 mobile phase eluting 15L, TLC points plate collect flavone reaction
Eluent part.
7. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 1, it is characterised in that：In step E, 20%~
The process of 70% methanol solution gradient elution is to be prepared using binary pump gradient, per 2min Fractional Collectionses, is examined by TLC points plate
Survey, collect the eluent of flavone reaction.
8. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 1, it is characterised in that：In step E, the solution
Analysis methanol concentration is 90%.
9. Flos Rosae Rugosae extraction and purification process for total flavonoids according to claim 2, it is characterised in that：The column chromatography polyamide
Consumption is 80~120 times of Flos Rosae Rugosae total flavones crude product gross weight, and during the wet method dress post, the volume ratio of dichloromethane and methanol is
10:1；The employing dichloromethane：Methanol is 10：1~1：It is 10 that 1 ratio carries out the process of gradient elution：1 mobile phase eluting
15L, 8：1 mobile phase eluting 10L, 6：1 mobile phase eluting 10L, 3：1 mobile phase eluting 10L, 1：1 mobile phase eluting 15L, TLC points
Plate collects the eluent part of flavone reaction.
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