CN107823233A - A kind of extracting method of the flavones based on angelica keiskei koidz - Google Patents
A kind of extracting method of the flavones based on angelica keiskei koidz Download PDFInfo
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- CN107823233A CN107823233A CN201711046763.XA CN201711046763A CN107823233A CN 107823233 A CN107823233 A CN 107823233A CN 201711046763 A CN201711046763 A CN 201711046763A CN 107823233 A CN107823233 A CN 107823233A
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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Abstract
The invention discloses a kind of extracting method of the flavones based on angelica keiskei koidz.This method includes tissue cultures, water planting, greenhouse gardening, extracting flavonoids.Extracting flavonoids process is divided into refluxing extraction, backflow plus microwave loss mechanisms, according to solid-liquid ratio 1:30‑1:The volume fraction ethanol of 40 ratios addition 50% 60%, soaks 90 120min, and 60 70 DEG C of 90min of refluxing extraction 60 are carried out after obtaining Flavonoid extract；After refluxing extraction the microwave digestion time be 1.5 2.5min, microwave digestion intensity be 360 480W.The present invention improves from the growth pattern of angelica keiskei koidz, processing is extracted optimization etc. and set about, there is provided the extracting method of the flavones of a set of complete angelica keiskei koidz, the flavones for solving current angelica keiskei koidz obtain the problems such as process efficiency is not high, purity is relatively low.
Description
Technical field
The invention belongs to agricultural production manufacture field, is related to a kind of extracting method of the flavones based on angelica keiskei koidz.
Background technology
Angelica keiskei koidz is composite family pseudo-ginseng platymiscium, also known as Gynura divaricata, Bai Dongfeng, beautiful loquat, asiatic toddalia root-bark, thick musculus cutaneus, chicken dish, big
Beef fat, white kind of amaranth etc., are distributed in Taiwan to south China, a southwestern generation, and happiness is born on the shaded place of humidity.Its rhizome is loose, meat, is in
Lumps, footpath 2-3cm, lark when fresh, more pieces of the raw stem eye in side.
Angelica keiskei koidz the whole province can be used as medicine, its acrid flavour, light；It is mild-natured, there is heat-clearing, relaxing muscles and tendons, hemostasis, dissolving stasis and other effects, can
Treat the functions such as pertussis, rheumatalgia, fracture, traumatic bleeding, hypoglycemic, pulmonary tuberculosis.Modern study shows that it has medicinal
And healthcare function, the abundant total flavonoid contained with it have close relationship.Flavonoids belongs to Plant Secondary Materials and writes product, is
Plurality of Chinese plays the material base of physiological action.At present, flavones acquisition research is more, from plant extract angle
Hand, such as solvent extraction.Conventional solvents extraction method has the problems such as recovery rate is low, extraction impurity is more.Special for angelica keiskei koidz
The extracting method of flavones, research is more single at present, it is impossible to well on the basis of cost is reduced, improve efficiency, maximum journey
Degree obtains high-purity flavones.
The content of the invention
It is an object of the invention to provide one kind to improve angelica keiskei koidz general flavone pick-up rate, and the flavones for solving current angelica keiskei koidz obtains
The problems such as process efficiency is not high, purity is relatively low.
To achieve the above object, the present invention uses following technical scheme：
The present invention provides a kind of extracting method of the flavones based on angelica keiskei koidz, and this method comprises the following steps：
(1) tissue cultures：Angelica keiskei koidz blade is taken under aseptic technique, to be connected to and change with vacuum side of blade as explant
In good MS inducing cultures, induction differentiation obtains callus, wherein, improvement MS inducing cultures are added with 1.5-2.2mg/L
6- benzyls aminoadenine (6-BA), 0.1-0.3mg/L methyl α-naphthyl acetate (NAA), 1.3-1.7mg/L polyvinylpyrrolidone
(PVP), pH5.5-6；After light culture 8-10 days, intensity of illumination is transferred in 1000-1500Lx culturing room, to keep humidity 80-
85%, cultivate 8-10 days；Squamous subculture 2-3 transfers in differential medium after generation, wherein, differential medium 0.2-0.4mg/
L methyl α-naphthyl acetate, the 1/2MS culture mediums of 2-5% mass fraction sucrose；Under 1500-2000Lx intensities of illumination, cultivate 20-30 days, obtain
Obtain test tube seedling；
(2) water planting：Test tube seedling is inoculated in water planting liquid, wherein, the MnSO added with 40-45mg/L4、6.0-7.0mg/
L ZnSO4；Under natural light, cultivate 8-10 days, obtain angelica keiskei koidz seedling；
(3) greenhouse gardening：According to turf：Vermiculite：Fine sand is by volume (3-5):(0.5-1.5):(0.5-1.5) prepares mixed
Close culture medium；By angelica keiskei koidz seedling cultivation in mixed culture medium, under natural light, after cultivating 5-8 days, according to 1500-2000kg/
Mu applies decomposed manure；Angelica keiskei koidz cauline leaf is harvested after 4-5 months；Ventilation in good time, irrigate；
(4) general flavone extracts：By angelica keiskei koidz cauline leaf natural air drying, shearing cauline leaf crushed 20-30 mesh to 2-3mm fritters
Sieve；According to solid-liquid ratio 1:30-1:40 ratios add 50%-60% volume fraction ethanol, soak 90-120min, obtain flavones leaching
Cream.
The method of above-mentioned raising angelica keiskei koidz general flavone pick-up rate, wherein, add (20-25) in water planting liquid：The Ca of (75-80)
(NO3)2(NH4)SO4, total nitrogen level is 3-4mmol/L.
The method that angelica keiskei koidz general flavone pick-up rate is carried on the back in above-mentioned raising in vain, wherein, addition 28-32g/L sucrose in water planting liquid
As carbon source.
The method for improving angelica keiskei koidz general flavone pick-up rate, wherein, the angelica keiskei koidz annual greenhouse gardening time is May and 10
Month, October and harvesting in May next year.
A kind of extracting method of above-mentioned flavones based on angelica keiskei koidz, wherein, after step (4), by Flavonoid extract and
Phase analysis is positioned in circulation reflux device together carries out refluxing extraction, under the conditions of 60-70 DEG C, refluxing extraction 3-4 times, and each 60-
90min。
A kind of extracting method of above-mentioned flavones based on angelica keiskei koidz, wherein, between refluxing extraction number interval, it will soak
Cream and phase analysis are positioned over power to extract 1.5-2.5min under 360-480W microwaves；Then, medicinal extract is entered together with material residue
Refluxing extraction process next time.
A kind of extracting method of above-mentioned flavones based on angelica keiskei koidz, wherein, after extraction terminates, medicinal extract is concentrated under reduced pressure
30-40min；Purified using polyamide Dynamic Adsorption concentrate, wherein, eluent is 70-75% ethanol, during elution
Between be 2-4BV/h.
A kind of extracting method of above-mentioned flavones based on angelica keiskei koidz, wherein, in polyamide purge process, resin medicinal material
Than for 1.5:1-2:1；Resin blade diameter length ratio is 1:8-1:10.
A kind of extracting method of above-mentioned flavones based on angelica keiskei koidz, wherein, loading pH remains 2.9-3.1；PH is cleaned to protect
Hold as 2-4.
Compared with prior art, a kind of extracting method of flavones based on angelica keiskei koidz of the invention has the following advantages that：
1. a kind of extracting method of flavones based on angelica keiskei koidz of the present invention, improved from angelica keiskei koidz growth pattern, processing carries
Optimization etc. is taken to set about, there is provided the method for a set of complete raising angelica keiskei koidz general flavone pick-up rate.
A kind of 2. extracting method of the flavones based on angelica keiskei koidz of the present invention, by needed for reasonable selection tissue culture procedures
Hormone, nutrient so that final test tube seedling general flavone content brings up to 3.9mg/g, and test tube sprout total flavone effective dose
Accumulating the raising to angelica keiskei koidz general flavone content after maturation has important facilitation.
3. a kind of extracting method of the flavones based on angelica keiskei koidz of the present invention, the nascent examination for creatively obtaining tissue cultures
Pipe seedling carries out the water planting hardening of 8-10 days, reasonable disposition Mn2+、Zn2+、Ca(NO3)2、(NH4)2SO4So that final ripe angelica keiskei koidz
General flavone content brings up to 11.8, and increase ratio is 640%.
4. a kind of extracting method of the flavones based on angelica keiskei koidz of the present invention, improving the base of angelica keiskei koidz plantation efficiency and quality
On plinth, also reasonable each Parameters variation in control angelica keiskei koidz extraction process, not only simple to operate but also relative to traditional recovery rate 6-
8% improves 2-4% percentage points, has important economic value to scale extraction production.Purified with reference to polyamide
Journey, whole angelica keiskei koidz general flavone pick-up rate can reach more than 11%, and purity is more than 35%.
Embodiment
To make the object, technical solutions and advantages of the present invention of greater clarity, below by embodiment, the present invention is entered
Row is further described.However, it should be understood that the specific embodiments described herein are merely illustrative of the present invention, and do not have to
In limitation the scope of the present invention.
Embodiment 1：
A kind of extracting method for flavones based on angelica keiskei koidz that embodiment 1 provides, comprises the following steps：
(1) culture medium is prepared：Prepare improvement MS inducing cultures：30g/L sucrose is added, agar content 8%, adjusts pH
For 5.5-6,1.5mg/L 6-BA, 0.1mg/L NAA, 1.3mg/L PVP are added；Prepare differential medium：2% mass point
Number sucrose, add 0.2mg/L NAA；Prepare mill water culture nutrient solution preparation：Nutrient composition includes following 28g/L sucrose, Ca
(NO3)20.07g/L；(NH4)SO40.2g/L；KH2PO4 1g/L、CaCl2·2H2O 5g/L, KCl 5g/L, MgSO42g/L；
H3BO30.046g/L；MnCl2·4H2O 0.04g/L；ZnSO4·7H2O 0.06g/L；CuSO4·5H2O 0.003g/L；
H2MoO4·H2O 0.001g/L, 1mL Fe-EDTA solution is added in every liter of nutrient solution, distilled water is prepared, and regulation pH is 5.8-
6.2。
(2) tissue cultures：In April, health is taken to obtain its young piece without insect pest angelica keiskei koidz, clean, cut blade edge, obtain
1*1cm explants；Under aseptic technique, angelica keiskei koidz explant is placed in inducing culture, light culture 8 days, keeps humidity
80%, after the generation of squamous subculture 2, it is transferred in differential medium, is cultivated 20 days under 2000Lx intensities of illumination, obtains test tube seedling.
(3) water planting：Above-mentioned test tube seedling is inoculated in mill water culture nutrient solution, under natural light, cultivated 10 days, obtains angelica keiskei koidz children
Seedling.
(4) greenhouse gardening：By turf, vermiculite and fine sand according to volume ratio 3:0.5:0.5 is configured to mixed culture medium, will be bright
Month careless seedling cultivation is in mixed culture medium, and natural optical culture is after 8 days, and decomposed manure is applied according to 2000kg/ mus, 2 months
Second of decomposed manure is applied according to 1500kg/ mus afterwards, is harvested after 3 months.
(5) extracting flavonoids：In October, angelica keiskei koidz cauline leaf is sheared, is cut after natural air drying to 3mm fritters, crushed 30
Mesh sieve；According to solid-liquid ratio 1:40 ratios add 50% volume fraction ethanol, soak 120min, obtain first time Flavonoid extract；Will
First time Flavonoid extract and phase analysis are positioned in circulation reflux device, and first time refluxing extraction, backflow are carried out under the conditions of 60 DEG C
90min；After first time refluxing extraction, medicinal extract and phase analysis are positioned over power to extract 2.5min under 360W microwaves；Microwave carries
Enter refluxing extraction next time after taking.After refluxing extraction 2 times, stop refluxing extraction and obtain medicinal extract, be filtered under diminished pressure 40min to crude drug
Measure 0.1g/L.
(6) flavones purifies：Polyamide is weighed according to 2 times of mass ratioes of medicinal material.Polyamide is boiled with ethanol 2 times,
Each 1h, wet method upper prop, it is 1 to obtain resin blade diameter length ratio:8.The concentrated extract pH3.0 that regulating step (5) obtains, with less than 4BV/
Concentrated extract is added dropwise in polyamide by h flow velocitys, and eluent, 3BV/h are used as by 70% ethanol of pH3.0 mass concentrations
It is eluted in spectrophotometer detection eluent and Flavonoid substances is not present.
Embodiment 2：
A kind of extracting method for flavones based on angelica keiskei koidz that embodiment 2 provides, comprises the following steps：
(1) culture medium is prepared：Prepare improvement MS inducing cultures：Add 35g/L sucrose, agar content 10%, regulation
PH is 5.5-6, adds 2.2mg/L 6-BA, 0.3mg/L NAA, 1.7mg/L PVP；Prepare differential medium：5% mass
Fraction sucrose, add 0.4mg/L NAA；Prepare mill water culture nutrient solution preparation：Nutrient composition includes following 32g/L sucrose, Ca
(NO3)20.06g/L；(NH4)SO40.2g/L；KH2PO4 1.2g/L、CaCl2·2H2O 4.5g/L, KCl 5.3g/L, MgSO4
2.2g/L；H3BO30.050g/L；MnCl2·4H2O 0.042g/L；ZnSO4·7H2O 0.058g/L；CuSO4·5H2O
0.002g/L；H2MoO4·H2O 0.002g/L, 1mL Fe-EDTA solution is added in every liter of nutrient solution, distilled water is prepared, regulation
PH is 5.8-6.2.
(2) tissue cultures：In April, health is taken to obtain its young piece without insect pest angelica keiskei koidz, clean, cut blade edge, obtain
1*1cm explants；Under aseptic technique, angelica keiskei koidz explant is placed in inducing culture, light culture 10 days, keeps humidity
85%, after the generation of squamous subculture 3, it is transferred in differential medium, is cultivated 30 days under 1500Lx intensities of illumination, obtains test tube seedling.
(3) water planting：Above-mentioned test tube seedling is inoculated in mill water culture nutrient solution, under natural light, cultivated 10 days, obtains angelica keiskei koidz children
Seedling.
(4) greenhouse gardening：By turf, vermiculite and fine sand according to volume ratio 5:1.5:1.5 are configured to mixed culture medium, will be bright
Month careless seedling cultivation is in mixed culture medium, and natural optical culture is after 5 days, and decomposed manure is applied according to 1500kg/ mus, 2 months
Second of decomposed manure is applied according to 1500kg/ mus afterwards, is harvested after 3 months.
(5) extracting flavonoids：In October, angelica keiskei koidz cauline leaf is sheared, is cut after natural air drying to 2mm fritters, crushed 20
Mesh sieve；According to solid-liquid ratio 1:30 ratios add 60% volume fraction ethanol, soak 90min, obtain first time Flavonoid extract；By
Flavonoid extract and phase analysis are positioned in circulation reflux device, and first time refluxing extraction, backflow are carried out under the conditions of 60 DEG C
60min；After first time refluxing extraction, medicinal extract and phase analysis are positioned over power to extract 1.5min under 480 microwaves；Microwave Extraction
Enter refluxing extraction next time afterwards.After refluxing extraction 2 times, stop refluxing extraction and obtain medicinal extract, be filtered under diminished pressure 30min to crude drug amount
0.1g/L。
(6) flavones purifies：Polyamide is weighed according to 2 times of mass ratioes of medicinal material.Polyamide is boiled with ethanol 2 times,
Each 1h, wet method upper prop, it is 1 to obtain resin blade diameter length ratio:10.Regulating step (5) obtain concentrated extract pH3.0, with less than
Concentrated extract is added dropwise in polyamide by 4BV/h flow velocitys, using pH3.0 mass concentrations as 75% ethanol as eluent,
2BV/h is eluted in spectrophotometer detection eluent and Flavonoid substances is not present.
Control group：
1. influence of the culture medium to the yield of flavone in angelica keiskei koidz
(1) influence of the inducing culture to the yield of flavone in angelica keiskei koidz
Preparing includes the improvement MS inducing cultures of different proportion hormone, as described in Table 1, in kind carries out induction training
Support, differentiation culture, using ponkan fourth as standard items after 30 days, general flavone amount in test tube seedling determined by ultraviolet specrophotometer,
As shown in table 2, in all improvement MS inducing cultures, 6-BA, NAA added with scope of the present invention, test tube sprout total flavone
Content is up to 3.6mg/g, as shown in table 1.
Influence of the different improvement MS inducing cultures of table 1 to test tube seedling general flavone content
(2) influence of the yield of flavone in differential medium angelica keiskei koidz
Preparing includes the 1/2MS differential mediums of different proportion NAA, sucrose, as described in Table 2, with 1.5mg/L, 0.1mg/
LNAA, 1.3mg/LPVP improvement MS inducing culture inductions are into callus, with above-mentioned 1/2MS differential mediums in the same terms
Lower differentiation culture, using ponkan fourth as standard items after 30 days, general flavone amount in test tube seedling is determined by ultraviolet specrophotometer, such as
Shown in table 2, under the conditions of all 1/2MS differential mediums, 6-BA, NAA added with scope of the present invention, test tube seedling is always yellow
Ketone content is up to 3.9mg/g, as shown in table 2.
Influence of the different 1/2MS differential mediums of table 2 to test tube seedling general flavone content
2. the influence of cultivating condition angelica keiskei koidz yield of flavone
(1)Mn2+、Zn2+Influence to the yield of flavone in angelica keiskei koidz
Inducing culture is used as using 1.5mg/L, 0.1mg/LNAA, 1.3mg/LPVP improvement MS；With 0.2mg/L, 2% mass
Fraction sucrose 1/2MS culture mediums obtain test tube seedling as differential medium.Different water planting liquid are prepared, are carried out with same culture conditions
Culture, is transferred to greenhouse and is planted May, harvesting in October, general flavone amount is determined with ultraviolet specrophotometer, such as table 3, MnSO4、
ZnSO4In scope provided by the invention, flavones content can reach 11.8mg/g, and exceed the scope, and flavones content is equal
Have and decline by a relatively large margin.
The Mn of table 32+、Zn2+Influence to angelica keiskei koidz general flavone content
(2) it is different to supply influence of the nitrogen level to angelica keiskei koidz yield of flavone
0.05g/L is prepared respectively：0.20g/L；0.06g/L:0.19g/L；0.07g/L:0.28g/L；0.09g/L:
0.26g/L；0.15g/L:0.60g/L；0.01g/L:0.04g/L mass ratio Ca (NO3)2：(NH4)2SO4；Other conditions are identical, with
Ultraviolet specrophotometer determines general flavone amount, such as table 4.
Table 4 is different to supply influence of the nitrogen level to angelica keiskei koidz general flavone content
3. extracting flavonoids procedure parameter com-parison and analysis
(1) influence of the Different Extraction Method to angelica keiskei koidz general flavone yield
The extraction examination of angelica keiskei koidz general flavone is carried out with microwave method, circumfluence method and microwave of the present invention+circumfluence method respectively
Test, by absorbance measuring, microwave method general flavone yield is 7.4%, circumfluence method general flavone yield is 9.1%, microwave+circumfluence method
Angelica keiskei koidz general flavone yield is 10.1%, it is seen that carries out angelica keiskei koidz general flavone extraction energy with microwave provided by the invention+circumfluence method
It is enough largely to improve general flavone yield.
(2) soak time influences on angelica keiskei koidz general flavone yield before extracting
With 10 minutes for grade, 30-180min is set respectively, ethanol soak time parallel test, entered with backflow+microwave method
Row angelica keiskei koidz general flavone recovery rate is tested, by absorbance measuring, as ethanol soak time extends, final general flavone yield from
3.4% brings up to the 9.8% of 100 minutes, and subsequent general flavone yield starts slowly to decline, and declines to general flavone yield during 180min
To 8.7%.
(3) concentration of alcohol influences on angelica keiskei koidz general flavone yield
It is respectively that 20%, 30%, 40%, 50%, 60%, 70%, 80% ethanol carries out the+microwave method that flows back with mass concentration
Carry out the experiment of angelica keiskei koidz general flavone recovery rate, general flavone yield is respectively 5.2%, 5.8%, 6.7%, 9.0%, 9.7%,
8.1%th, 7.3%.
(4) solid-liquid ratio influences on angelica keiskei koidz general flavone yield
1 is set respectively:10、1:20、1:30、1:40、1:50、1:60、1:70、1:80 solid-liquid ratios carry out the+microwave method that flows back
Carry out the experiment of angelica keiskei koidz general flavone recovery rate, general flavone yield is respectively 3.1%, 6.8%, 8.9%, 9.4%, 9.9%,
10.2%th, 10.1%, 9.8%.
(5) extraction time influences on angelica keiskei koidz general flavone yield
Using 1+1 microwave of backflow as time cycle, 1,2,3,4,5,6 backflow+microwave method is carried out respectively and carries out angelica keiskei koidz
General flavone recovery rate is tested, and general flavone yield is respectively 8.1%, 9.3%, 9.9%, 10.3%, 10.1%, 10.2%, explanation
After extraction time is more than 4 times, general flavone yield does not change.
(6) microwave treatment time influences on the recovery rate of the flavones in angelica keiskei koidz
Using 0.5min as grade, respectively set 0.5min, 1.0min, 1.5min, 2.0min, 2.5min, 3.0min,
3.5min the microwave digestion time, the extracting flavonoids in 3 wheel backflow+microwave method angelica keiskei koidzs are carried out, on every grade microwave digestion time
Often wheel extraction time is identical, and general flavone yield is respectively 8.3%, 8.8%, 9.4%, 9.8%, 9.9%, 9.4%, 8.6%.
It is recognised that the illustrative embodiments that above-described embodiment uses only for explanation inventive principle, but this hair
Bright to be not limited only to this, those skilled in the art can make various improvement and change in the case where not departing from real situation of the present invention, this
A little improvement and change fall within protection scope of the present invention.
Claims (9)
1. a kind of extracting method of the flavones based on angelica keiskei koidz, it is characterised in that this method comprises the following steps：
(1) tissue cultures：Angelica keiskei koidz blade is taken under aseptic technique, improvement MS to be connected to vacuum side of blade as explant
In inducing culture, induction differentiation obtains callus, wherein, improvement MS inducing cultures are added with 1.5-2.2mg/L 6-
Benzyl aminoadenine, 0.1-0.3mg/L methyl α-naphthyl acetate, 1.3-1.7mg/L polyvinylpyrrolidone, pH5.5-6；Light culture 8-
After 10 days, intensity of illumination is transferred in 1000-1500Lx culturing room, to keep humidity 80-85%, is cultivated 8-10 days；Squamous subculture
2-3 transfers in differential medium after generation, wherein, differential medium is 0.2-0.4mg/L methyl α-naphthyl acetate, 2-5% mass fraction
The 1/2MS culture mediums of sucrose；Under 1500-2000Lx intensities of illumination, cultivate 20-30 days, obtain test tube seedling；
(2) water planting：Test tube seedling is inoculated in water planting liquid, wherein, the MnSO added with 40-45mg/L4, 6.0-7.0mg/L Zn
SO4；Under natural light, cultivate 8-10 days, obtain angelica keiskei koidz seedling；
(3) greenhouse gardening：According to turf：Vermiculite：Fine sand is by volume (3-5):(0.5-1.5):(0.5-1.5) prepares mixing training
Support base；By angelica keiskei koidz seedling cultivation in mixed culture medium, under natural light, after cultivating 5-8 days, applied according to 1500-2000kg/ mus
Enter decomposed manure；Angelica keiskei koidz cauline leaf is harvested after 4-5 months；Ventilation in good time, irrigate；
(4) extracting flavonoids：By angelica keiskei koidz cauline leaf natural air drying, shearing cauline leaf crushed 20-30 mesh sieves to 2-3mm fritters；According to
Solid-liquid ratio 1:30-1:40 ratios add 50%-60% volume fraction ethanol, soak 90-120min, obtain Flavonoid extract.
2. the extracting method based on the flavones based on angelica keiskei koidz described in claim 1, it is characterised in that：Matter is added in water planting liquid
Amount is than (20-25)：Ca (the NO of (75-80)3)2(NH4)2SO4, total nitrogen level is 0.25-0.35g/L.
3. the extracting method based on the flavones based on angelica keiskei koidz described in claim 1, it is characterised in that：Added in water planting liquid
28-32g/L sucrose is as carbon source.
4. the extracting method based on the flavones based on angelica keiskei koidz described in claim 1, it is characterised in that：The annual greenhouse of angelica keiskei koidz
Implantation time is May and October, is gathered in October and May next year.
5. the extracting method based on the flavones based on angelica keiskei koidz described in claim 4, it is characterised in that：, will after step (4)
Flavonoid extract and phase analysis are positioned in circulation reflux device together carries out refluxing extraction, under the conditions of 60-70 DEG C, refluxing extraction 3-
4 times, each 60-90min.
6. the extracting method based on the flavones based on angelica keiskei koidz described in claim 5, it is characterised in that：In refluxing extraction number
Between interval, medicinal extract and phase analysis are positioned over power to extract 1.5-2.5min under 360-480W microwaves；Then, by medicinal extract together with
Material residue enters refluxing extraction process next time together.
7. the extracting method based on the flavones based on angelica keiskei koidz described in claim 5 or 6, it is characterised in that：After extraction terminates,
Medicinal extract is concentrated under reduced pressure 30-40min, guarantee contains crude drug amount >=0.1g/ml；Carried out using polyamide Dynamic Adsorption concentrate
Purifying, wherein, eluent is 70-75% mass concentration ethanol, sample-adding, elution speed 2-4BV/h.
8. the extracting method based on the flavones based on angelica keiskei koidz described in claim 7, it is characterised in that：Polyamide purifies
During, resin medicinal material ratio is 1.5:1-2:1；Resin blade diameter length ratio is 1:8-1:10.
9. the extracting method based on the flavones based on angelica keiskei koidz described in claim 8, it is characterised in that：Loading pH is remained
2.9-3.1；Cleaning pH remains 2-4.
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| CN102246694A (en) * | 2011-05-05 | 2011-11-23 | 湖南农业大学 | Tissue culture method of gynura divaricata |
| CN106359082A (en) * | 2016-08-19 | 2017-02-01 | 黑龙江省科学院大庆分院 | Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet |
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|---|---|---|---|---|
| CN102246694A (en) * | 2011-05-05 | 2011-11-23 | 湖南农业大学 | Tissue culture method of gynura divaricata |
| CN106359082A (en) * | 2016-08-19 | 2017-02-01 | 黑龙江省科学院大庆分院 | Tissue culture method with effect of improving general flavone content of gynura divaricata test-tube plantlet |
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