CN107896988A - A kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae - Google Patents

A kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae Download PDF

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CN107896988A
CN107896988A CN201711048769.0A CN201711048769A CN107896988A CN 107896988 A CN107896988 A CN 107896988A CN 201711048769 A CN201711048769 A CN 201711048769A CN 107896988 A CN107896988 A CN 107896988A
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radix
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rate
gynurae divaricatae
rhizoma gynurae
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艾永冠
韦玲
李真真
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China Anhui Biological Technology Co Ltd
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Abstract

The invention discloses a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae.Compared with prior art, a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae of the invention, on the basis of Radix et Rhizoma Gynurae divaricatae plantation efficiency and quality is improved, rationally controls each Parameters variation in Radix et Rhizoma Gynurae divaricatae extraction process.It is not only easy to operate but also improve 2 4% percentage points relative to traditional recovery rate 6 8%, there is important economic value to scale extraction production.With reference to polyamide purge process, whole white-backed pseudo-ginseng total flavonoid pick-up rate can reach more than 11%, and purity is more than 35%.

Description

A kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae
Technical field
The invention belongs to agricultural production manufacture field, is related to a kind of side for being used to improve flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae Method.
Background technology
Radix et Rhizoma Gynurae divaricatae 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, China to south China, a southwestern generation, happiness and are born on the shaded place of humidity.Its rhizome hypertrophy, Meat, in lumps, footpath 2-3cm, lark when fresh, more pieces of the raw stem eye in side.
Radix et Rhizoma Gynurae divaricatae 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, energy Enough treat the functions such as pertussis, rheumatalgia, fracture, traumatic bleeding, hypoglycemic, pulmonary tuberculosis.Modern research shows that its medicine for having With and healthcare function, the abundant total flavonoid contained with it there is close relationship.Flavonoids belongs to Plant Secondary Materials and writes product, It is the material base that plurality of Chinese plays physiological action.At present, flavones acquisition research is more, is generally from plant extract angle Set about, such as solvent extraction.Conventional solvents extraction method has the problems such as recovery rate is low, extraction impurity is more.It is special for Radix et Rhizoma Gynurae divaricatae The preparation method of different general flavone, research is more single at present, it is impossible to well on the basis of cost is reduced, improve efficiency, At utmost obtain high-purity general flavone.
The content of the invention
The object of the present invention is to provide a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae, solves the white back of the body at present Radix notoginseng flavones obtain 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 method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae, and this method comprises the following steps：
(1) tissue cultures：Radix et Rhizoma Gynurae divaricatae blade is taken under aseptic technique, to be connected to as explant with vacuum side of blade Improve in MS inducing cultures, induction differentiation obtains callus, and for light culture after 8-10 days, it is 1000- to be transferred to intensity of illumination In 1500Lx culturing room, humidity 80-85% is kept, is cultivated 8-10 days；Squamous subculture 2-3 transfers in differential medium after generation, Wherein, differential medium is methyl α-naphthyl acetate, the 1/2MS culture medium of 2-5% mass fraction sucrose of 0.2-0.4mg/L；1500- Under 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- The ZnSO of 7.0mg/L4；Under natural light, cultivate 8-10 days, obtain Radix et Rhizoma Gynurae divaricatae seedling；
(3) greenhouse gardening：According to turf：Vermiculite：Fine sand is by volume (3-5):(0.5-1.5):(0.5-1.5) is prepared Mixed culture medium；By Radix et Rhizoma Gynurae divaricatae seedling cultivation in mixed culture medium, under natural light, after cultivating 5-8 days, according to 1500- 2000kg/ mus of application decomposed manures；Divaricate velvetplant leaf is harvested after 4-5 months；Ventilation in due course, irrigate；
(4) general flavone extracts：By divaricate velvetplant leaf natural air drying, shearing cauline leaf crushed 20-30 to 2-3mm fritters Mesh sieve；According to solid-liquid ratio 1:30-1:40 ratios add 50%-60% volume fraction ethanol, soak 90-120min, obtain flavones Medicinal extract.
As the further optimization of such scheme, wherein, improvement MS inducing cultures are added with the 6- of 1.5-2.2mg/L Benzyl aminoadenine (6-BA), the methyl α-naphthyl acetate (NAA) of 0.1-0.3mg/L, the polyvinylpyrrolidone of 1.3-1.7mg/L (PVP), pH5.5-6.
The method of above-mentioned raising white-backed pseudo-ginseng total flavonoid pick-up rate, wherein, add (20-25) in water planting liquid：(75-80) Ca (NO3)2(NH4)SO4, total nitrogen level is 3-4mmol/L.
The method that white-backed pseudo-ginseng total flavonoid pick-up rate is carried on the back in above-mentioned raising in vain, wherein, the sugarcane of addition 28-32g/L in water planting liquid Sugar is used as carbon source.
The method for improving white-backed pseudo-ginseng total flavonoid pick-up rate, wherein, the Radix et Rhizoma Gynurae divaricatae annual greenhouse gardening time is 5 The moon and October, October and harvesting in May next year.
The method of above-mentioned raising white-backed pseudo-ginseng total flavonoid pick-up rate, wherein, after step (4), by Flavonoid extract and leaching Slag 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。
The method of above-mentioned raising white-backed pseudo-ginseng total flavonoid pick-up rate, wherein, between refluxing extraction number interval, by medicinal extract Power is positioned over phase analysis to extract 1.5-2.5min under 360-480W microwaves；Then, under medicinal extract is entered together with material residue Refluxing extraction process.
The method of above-mentioned raising white-backed pseudo-ginseng total flavonoid pick-up rate, wherein, after extraction, medicinal extract is concentrated under reduced pressure 30- 40min；Purified using polyamide Dynamic Adsorption concentrate, wherein, eluent is 70-75% ethanol, elution time For 2-4BV/h.
The method of above-mentioned raising white-backed pseudo-ginseng total flavonoid pick-up rate, 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.
The method of above-mentioned raising white-backed pseudo-ginseng total flavonoid pick-up rate, wherein, loading pH remains 2.9-3.1；PH is cleaned to protect Hold as 2-4.
Compared with prior art, a kind of method for being used to improve flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae of the invention is with as follows Advantage：
1. a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae of the invention, changes from Radix et Rhizoma Gynurae divaricatae growth pattern Set about into, processing extraction optimization etc., there is provided the method for a set of complete raising Radix et Rhizoma Gynurae divaricatae flavones pick-up rate.
A kind of 2. method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae of the invention, by reasonably selecting tissue cultures mistake Hormone, nutrient needed for journey so that final test tube seedling general flavone content brings up to 3.9mg/g, and test tube sprout total flavone Raising of a effective amount of accumulation to white-backed pseudo-ginseng total flavonoid content after maturation has important facilitation.
3. a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae of the invention, creatively obtains tissue cultures Nascent test tube seedling carry out the water planting hardening of 8-10 days, reasonable disposition Mn2+、Zn2+、Ca(NO3)2、(NH4)2SO4So that finally into Ripe white-backed pseudo-ginseng total flavonoid content brings up to 11.8, and increase ratio is 640%.
4. a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae of the invention, is improving Radix et Rhizoma Gynurae divaricatae plantation efficiency On the basis of quality, also rationally control Radix et Rhizoma Gynurae divaricatae extraction process in each Parameters variation, it is not only easy to operate and relative to Traditional recovery rate 6-8% improves 2-4% percentage points, has important economic value to scale extraction production.With reference to polyamides Polyimide resin purge process, whole white-backed pseudo-ginseng total flavonoid pick-up rate can reach more than 11%, and purity is more than 35%.
Embodiment
With reference to specific embodiment to the present invention.
Embodiment 1：
(1) culture medium is prepared：Prepare improvement MS inducing cultures：30g/L sucrose is added, agar content 8%, adjusts pH For 5.5-6, the 6-BA, the PVP of the NAA of 0.1mg/L, 1.3mg/L of 1.5mg/L are added；Prepare differential medium：2% mass point Number sucrose, adds the NAA of 0.2mg/L；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, and distilled water is prepared, and adjusting pH is 5.8- 6.2。
(2) tissue cultures：In April, take health to obtain its young piece, cleaning, cuts blade edge, obtain without insect pest Radix et Rhizoma Gynurae divaricatae Obtain 1*1cm explants；Under aseptic technique, Radix et Rhizoma Gynurae divaricatae explant is placed in inducing culture, light culture 8 days, kept Humidity 80%, after 2 generation of squamous subculture, 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, cultivates 10 days, obtains Radix et Rhizoma Gynurae divaricatae 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 white Pseudo-ginseng seedling cultivation is carried on the back in mixed culture medium, natural optical culture is after 8 days, and decomposed manures are applied according to 2000kg/ mu, 2 According to second of decomposed manure of 1500kg/ mus of applications after month, harvested after 3 months.
(5) general flavone extracts：In October, shear divaricate velvetplant leaf, is cut to 3mm fritters, crushed after natural air drying Cross 30 mesh sieves；According to solid-liquid ratio 1:40 ratios add 50% volume fraction ethanol, soak 120min, obtain the leaching of first time flavones Cream；First time Flavonoid extract and phase analysis are positioned in circulation reflux device, first time refluxing extraction is carried out under the conditions of 60 DEG C, is returned Flow 90min；After first time refluxing extraction, medicinal extract and phase analysis are positioned over power to extract 2.5min under 360W microwaves；Microwave Enter refluxing extraction next time after extraction.After refluxing extraction 2 times, stop refluxing extraction and obtain medicinal extract, be filtered under diminished pressure 40min to life Dose 0.1g/L.
(6) general flavone purifies：Polyamide is weighed according to 2 times of mass ratioes of medicinal material.Polyamide 2 is boiled with ethanol Secondary, each 1h, wet method upper prop, it is 1 to obtain resin blade diameter length ratio:8.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 70% ethanol as eluent, 3BV/h is eluted in spectrophotometer detection eluent and Flavonoid substances is not present.
Embodiment 2：
(1) culture medium is prepared：Prepare improvement MS inducing cultures：35g/L sucrose is added, agar content 10%, is adjusted PH is 5.5-6, adds the 6-BA of 2.2mg/L, the PVP of the NAA of 0.3mg/L, 1.7mg/L；Prepare differential medium：5% mass Fraction sucrose, adds the NAA of 0.4mg/L；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, and distilled water is prepared, and is adjusted PH is 5.8-6.2.
(2) tissue cultures：In April, take health to obtain its young piece, cleaning, cuts blade edge, obtain without insect pest Radix et Rhizoma Gynurae divaricatae Obtain 1*1cm explants；Under aseptic technique, Radix et Rhizoma Gynurae divaricatae explant is placed in inducing culture, light culture 10 days, protected Humidity 85% is held, after 3 generation of squamous subculture, 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, cultivates 10 days, obtains Radix et Rhizoma Gynurae divaricatae 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 white Pseudo-ginseng seedling cultivation is carried on the back in mixed culture medium, natural optical culture is after 5 days, and decomposed manures are applied according to 1500kg/ mu, 2 According to second of decomposed manure of 1500kg/ mus of applications after month, harvested after 3 months.
(5) general flavone extracts：In October, shear divaricate velvetplant leaf, is cut to 2mm fritters, crushed after natural air drying Cross 20 mesh sieves；According to solid-liquid ratio 1:30 ratios add 60% volume fraction ethanol, soak 90min, obtain first time Flavonoid extract； First time Flavonoid extract and phase analysis are positioned in circulation reflux device, first time refluxing extraction, reflux 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) general flavone purifies：Polyamide is weighed according to 2 times of mass ratioes of medicinal material.Polyamide 2 is boiled with ethanol Secondary, each 1h, wet method upper prop, it is 1 to obtain resin blade diameter length ratio:10.The concentrated extract pH3.0 that regulating step (5) obtains, with small Concentrated extract is added dropwise in polyamide in 4BV/h flow velocitys, elution is used as by 75% ethanol of pH3.0 mass concentrations Liquid, 2BV/h are eluted in spectrophotometer detection eluent and Flavonoid substances are not present.
Control group：
1. influence of the culture medium to Radix et Rhizoma Gynurae divaricatae yield of flavone
(1) influence of the inducing culture to Radix et Rhizoma Gynurae divaricatae yield of flavone
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 measured 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 differential medium Radix et Rhizoma Gynurae divaricatae yield of flavone
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, measures general flavone amount in test tube seedling, such as by ultraviolet specrophotometer 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 difference 1/2MS differential mediums of table 2 to test tube seedling general flavone content
2. the influence of cultivating condition Radix et Rhizoma Gynurae divaricatae yield of flavone
(1)Mn2+、Zn2+Influence to Radix et Rhizoma Gynurae divaricatae yield of flavone
Inducing culture is used as using 1.5mg/L, 0.1mg/L NAA, 1.3mg/L PVP 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, with same culture conditions To be cultivated, May is transferred to greenhouse and is planted, and harvesting in October, general flavone amount is measured 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 contains Amount has to be declined by a relatively large margin.
3 Mn of table2+、Zn2+Influence to white-backed pseudo-ginseng total flavonoid content
(2) it is different to supply influence of the nitrogen level to Radix et Rhizoma Gynurae divaricatae yield of flavone
0.05g/L is prepared respectively：0.20g/L；0.06g/L:0.19g/L；0.07g/L:0.28g/L；0.09 g/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 measures general flavone amount, such as table 4.
Table 4 is different to supply influence of the nitrogen level to white-backed pseudo-ginseng total flavonoid content
3. the analysis of extracting flavonoids procedure parameter is compared
(1) influence of the Different Extraction Method to white-backed pseudo-ginseng total flavonoid yield
White-backed pseudo-ginseng total flavonoid extraction is carried out with microwave method, circumfluence method and microwave of the present invention+circumfluence method respectively Experiment, is measured by absorbance, and microwave method general flavone yield is 7.4%, circumfluence method general flavone yield is 9.1%, microwave+return Stream method white-backed pseudo-ginseng total flavonoid yield is 10.1%, it is seen that it is always yellow to carry out Radix et Rhizoma Gynurae divaricatae with microwave provided by the invention+circumfluence method Ketone extraction can largely improve general flavone yield.
(2) soaking time influences white-backed pseudo-ginseng total flavonoid yield before extracting
With 10 minutes for grade, 30-180min is set respectively, ethanol soaking time parallel test, with reflux+microwave method into Row white-backed pseudo-ginseng total flavonoid recovery rate is tested, and is measured by absorbance, as ethanol soaking time extends, final general flavone yield The 9.8% of 100 minutes is brought up to from 3.4%, subsequent general flavone yield starts slowly to decline, under general flavone yield during 180min It is down to 8.7%.
(3) concentration of alcohol influences white-backed pseudo-ginseng total flavonoid yield
Respectively+the microwave method that flows back is carried out for 20%, 30%, 40%, 50%, 60%, 70%, 80% ethanol with mass concentration Carry out white-backed pseudo-ginseng total flavonoid recovery rate experiment, 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 white-backed pseudo-ginseng total flavonoid 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 white-backed pseudo-ginseng total flavonoid recovery rate experiment, 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 white-backed pseudo-ginseng total flavonoid yield
Using 1+1 microwave of reflux as time cycle, 1,2,3,4,5,6 reflux+microwave method is carried out respectively and carries out the white back of the body three Seven general flavone recovery rates are tested, and general flavone yield is respectively 8.1%, 9.3%, 9.9%, 10.3%, 10.1%, 10.2%, is said It is bright when extraction time more than 4 times after, general flavone yield does not change.
(6) microwave treatment time influences white-backed pseudo-ginseng total flavonoid recovery rate
Using 0.5min as grade, respectively set 0.5min, 1.0min, 1.5min, 2.0min, 2.5min, 3.0min, The 3.5min microwave digestion times, carry out 3 wheel reflux+microwave method white-backed pseudo-ginseng total flavonoid extraction, 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 (10)

  1. A kind of 1. method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae, it is characterised in that this method comprises the following steps：
    (1) tissue cultures：Radix et Rhizoma Gynurae divaricatae blade is taken under aseptic technique, improvement to be connected to vacuum side of blade as explant In MS inducing cultures, induction differentiation obtains callus, and for light culture after 8-10 days, it is 1000-1500Lx to be transferred to intensity of illumination In culturing room, humidity 80-85% is kept, is cultivated 8-10 days；Squamous subculture 2-3 transfers in differential medium after generation, wherein, point Change methyl α-naphthyl acetate, the 1/2MS culture mediums of 2-5% mass fraction sucrose that culture medium is 0.2-0.4mg/L；1500-2000Lx illumination Under intensity, 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 Radix et Rhizoma Gynurae divaricatae 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 Radix et Rhizoma Gynurae divaricatae seedling cultivation in mixed culture medium, under natural light, after cultivating 5-8 days, according to 1500-2000kg/ mus Apply decomposed manure；Divaricate velvetplant leaf is harvested after 4-5 months；Ventilation in due course, irrigate；
    (4) extracting flavonoids：By divaricate velvetplant leaf natural air drying, shearing cauline leaf crushed 20-30 mesh sieves to 2-3mm fritters；Press Take care of liquor ratio 1:30-1:40 ratios add 50%-60% volume fraction ethanol, soak 90-120min, obtain Flavonoid extract.
  2. 2. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 1, it is characterised in that：Water Train and mass ratio (20-25) is added in liquid：Ca (the NO of (75-80)3)2(NH4)2SO4, total nitrogen level is 0.25-0.35g/L.
  3. 3. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 1, it is characterised in that：Water Train and the sucrose of 28-32g/L is added in liquid as carbon source.
  4. 4. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 1, it is characterised in that：In vain The back of the body pseudo-ginseng annual greenhouse gardening time is May and October, is gathered in October and May next year.
  5. 5. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 4, it is characterised in that：Step Suddenly after (4), Flavonoid extract and phase analysis is positioned in circulation reflux device together and carry out refluxing extraction, 60-70 DEG C of condition Under, refluxing extraction 3-4 times, each 60-90min.
  6. 6. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 5, it is characterised in that： Between refluxing extraction number interval, medicinal extract and phase analysis are positioned over power to extract 1.5-2.5min under 360-480W microwaves；With Afterwards, medicinal extract is entered into refluxing extraction process next time together with material residue.
  7. 7. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 5 or 6, its feature exists In：After extraction, medicinal extract is concentrated under reduced pressure 30-40min, guarantee contains crude drug amount >=0.1g/ml；Using polyamide dynamic Adsorption Concentration liquid is purified, wherein, eluent is 70-75% mass concentration ethanol, sample-adding, elution speed 2-4BV/h.
  8. 8. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 7, it is characterised in that：It is poly- In amide resin purge process, resin medicinal material ratio is 1.5:1-2:1；Resin blade diameter length ratio is 1:8-1:10.
  9. 9. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 8, it is characterised in that：On Sample pH remains 2.9-3.1；Cleaning pH remains 2-4.
  10. 10. based on a kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae described in claim 1, it is characterised in that： Improve the 6- benzyl aminoadenines that MS inducing cultures are added with 1.5-2.2mg/L, the methyl α-naphthyl acetate of 0.1-0.3mg/L, 1.3- The polyvinylpyrrolidone of 1.7mg/L, pH5.5-6.
CN201711048769.0A 2017-10-31 2017-10-31 A kind of method for improving flavones pick-up rate in Radix et Rhizoma Gynurae divaricatae Pending CN107896988A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111919694A (en) * 2020-08-21 2020-11-13 内蒙古大学 Hydroponic solution and cultivation method of high-quality radix astragali thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111919694A (en) * 2020-08-21 2020-11-13 内蒙古大学 Hydroponic solution and cultivation method of high-quality radix astragali thereof

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