CN101735292A - Production process for extracting, separating and purifying potentilla flavone - Google Patents
Production process for extracting, separating and purifying potentilla flavone Download PDFInfo
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- CN101735292A CN101735292A CN200910244861A CN200910244861A CN101735292A CN 101735292 A CN101735292 A CN 101735292A CN 200910244861 A CN200910244861 A CN 200910244861A CN 200910244861 A CN200910244861 A CN 200910244861A CN 101735292 A CN101735292 A CN 101735292A
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Abstract
The invention discloses a process for extracting, separating and purifying potentilla flavones, namely a chemical composition with antidiabetic effect in Chinese medicament potentilla, and belongs to the technology of extracting and separating active ingredients of Chinese medicaments. The compound has the following structure; and the process for extracting, separating and purifying the compound comprises the following steps: grinding the potentilla, adding 60 percent ethanol, heating, refluxing and extracting for three times in a solid-liquid ratio of 8:6:6, filtering, combining filter liquor, and evaporating and concentrating under reduced pressure to obtain an extract substance; dissolving the extract substance with 30 mass percent ethanol of which the weight is 10 times that of the extract substance, centrifuging at a rate of 3,000r/min, separating supernate with a 20- to 60-mesh AB-8 macroporous resin absorption column, making a sample volume 10BV (10 times of the column volume) and adsorption flow rate and elusion flow rate 2BV/h, eluting with 3BV of 30 percent ethanol and 5BV of 60 percent ethanol in turn, collecting eluent of the 60 percent ethanol, concentrating under reduced pressure until no alcohol smell exists, and obtaining a mixed suspension A; dissolving the mixed suspension with equal volume of 30 percent ethanol, centrifuging, separating supernate with 80- to 100-mesh polyamide chromatographic column, making a sample volume 10BV and adsorption and elusion flow rate 2BV/h, eluting with 3BV of 30 percent ethanol and 5BV of 60 percent ethanol in turn, collecting eluent of the 60 percent ethanol, concentrating under reduced pressure until no alcohol smell exists, and obtaining a mixed suspension B; centrifuging the mixed suspension B, purifying centrifuged precipitate with a high speed countercurrent chromatography, wherein in a solvent system, a ratio (volume ratio) of methylene dichloride to methanol to water is 5 to 4 to 2.
Description
Technical field
The present invention relates to a kind of extraction separation and purifying production process of tormentic anti-diabetic effective constituent-potentilla flavone, the extraction and the separation field of pharmaceutically active ingredient in belonging to.
Background technology
The Chinese medicine tormentic has another name called and turns over Chinese cabbage, Root of Chinese Pulsatilla, and Herba Potentillae Chinensis, Radix Rubi Parvifolii is the herb of the Rosaceae (Rosaceae) plant tormentic Potentilla chinesis Ser..This laboratory is according to the clinical application of Chinese medicine tormentic, with Chemistry for Chinese Traditional Medicine in conjunction with the pharmacology means, from the Chinese medicine tormentic isolation identification a compound-potentilla flavone (following formula) with antidiabetic effect, its drug effect and N1,N1-Dimethylbiguanide are suitable, have high clinical value.The present invention is the extraction separation and the purifying production process of potentilla flavone.
The structure of potentilla flavone
Extracting method commonly used in the Chinese medicine production at present is an ethanol-water solution heating and refluxing extraction method, but at different medicinal materials and target compound, required condition is different, as ethanolic soln concentration, extraction time, extraction time etc.Need utilize single factor or orthogonal test to investigate actual conditions.
The macroporous adsorbent resin chromatography method is a separation method commonly used during Chinese medicine is produced.But according to the physico-chemical property of target compound, optimal separation chromatograph packing material and separation condition all have difference, need investigate by various factors and determine.
The present invention in extraction process, has investigated the influence of extracting solvent strength, extraction time, extraction time and different feed liquid comparison potentilla flavone extraction yield and stability, and has determined optimum extraction process by orthogonal test according to the chemical technology research method; In separating technology, macroporous adsorbent resin (2 kinds of models) and polymeric amide have been investigated, determined the separation chromatography filler by Static Adsorption, dynamic adsorption and resolution factor, investigated of the influence of factors such as leakage plot and elution curve, absorption flow velocity and elution flow rate once more to the potentilla flavone separating effect, and the isolating effect of two dimensional chromatography, and further investigated the influence of last sample liquor ratio, chromatographic column blade diameter length ratio to separating effect, established optimal separation technology;
By extracting and separating, still be difficult to guarantee to obtain purity higher effective composition, also need to select suitable purification process, with separated material purifying.The present invention is to carry out purifying at a lot of new isolation technique-high speed adverse current chromatogram of application aspect the Separation of Natural Products purifying to potentilla flavone in recent years.(High-Speed Counter CurrentChromatography HSCCC) is a kind of liquid luquid partition chromatography isolation technique of continuous high-efficient to high speed adverse current chromatogram.This technology is not owing to need solid support, the separation of material to realize according to the difference of its partition ratio in two-phase, thereby avoided being particularly suitable for the separation of natural bioactive ingredients because of sample loss that irreversible adsorption causes, inactivation, sex change etc.In the HSCCC purifying process, two kinds of solvent systems (petroleum ether-ethyl acetate-methanol-waters have been investigated, methylene chloride-methanol-water) and the different solvents ratio to the influence of purification effect, the technology of having established methylene chloride-methanol-water solvent system (5: 4: 2) purifying potentilla flavone at last.
Summary of the invention
The object of the present invention is to provide a kind of production technique of extraction separation Chinese medicine tormentic effective constituent potentilla flavone, for the exploitation of new Chinese medicine provides a kind of extraction efficiency height, product purity is high and safe effective constituent production technique.
The present invention is realized by following extraction, separation and purification technique scheme, be it is characterized in that:
(1) extraction process: with the tormentic seasoning of gathering, be ground into brachyplast less than 0.5 centimetre, in medicinal material and 1: 8 ratio of aqueous ethanolic solution mass ratio, spend the night with 8 times of amount mass concentration 60% alcohol immersion, heating and refluxing extraction 1.5 hours, filter, measured mass concentration 60% alcohol reflux 1.5 hours with 6 times then, filter; And then, merge No. three times extracting solution with 6 times of amount mass concentration 60% alcohol reflux 1.5 hours, be condensed into fluid extract.
(2) separating technology: the fluid extract that step (1) is obtained, according to weight ratio, mass concentration 30% dissolve with ethanol with 10 times of amounts, fully stir, the centrifugal 10min of 3000r/min, get supernatant liquor, separate with 20-60 order AB-8 macroporous adsorptive resins, 10BV (10 times of column volumes) goes up sample, and the absorption flow velocity is 2BV/h, then successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h, collect 60% ethanol eluate, being evaporated to does not have the alcohol flavor, obtains a suspension A; Then with equal-volume 30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min gets supernatant liquor and separates the last sample of 10BV with 80-100 order polymeric amide chromatographic column, the absorption flow velocity is 2BV/h, successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h collects 60% ethanol eluate then, being evaporated to does not have the alcohol flavor, obtains a suspendible aqueous solution B.
(3) purifying process: the suspension B that step (2) obtains is centrifugal, the centrifugal 10min of 3000r/min, centrifugation is with preparation high-speed counter-current chromatograph purifying: solvent systems (volume ratio): methylene dichloride: methyl alcohol: water=5: 4: 2 below is stationary phase mutually, on be moving phase mutually, flow velocity 2ml/min, counter-rotating separates, rotating speed 800rpm, 25 ℃ of temperature, detect wavelength 254nm, each sample size 500mg.Collect flow point: 13min begins to collect behind the beginning appearance time, collects the wash-out flow point of 20min time period.The collection flow point is removed organic solvent through concentrating under reduced pressure and is got an aqueous suspension, and a centrifugal yellow mercury oxide with a small amount of 30% ethanolic soln rinse 2 times, promptly gets the potentilla flavone elaboration.
The anti-diabetic effective constituent potentilla flavone of making by this law can be separately or combine with suitable vehicle etc., makes oral dosage form or non-oral dosage form (injection, sprays, patch etc.) according to ordinary method, is used to prepare the medicine for the treatment of diabetes.
The inventor is by research for many years, develops a whole set of by the tormentic plant preparation technology as potentilla flavone as described in the following formula I, and compared with prior art, the present invention has following remarkable difference and beneficial effect:
1, the complete technical scheme by tormentic plant preparation I compound was not disclosed in the prior art;
2, adopt macroporous resin or polyamide column usually at separating step in the prior art, seldom have, especially do not occur being used for the isolating technical scheme of potentilla flavone after the two combination with the two bonded scheme;
3, the technical scheme of high speed adverse current chromatogram purifying potentilla flavone of no use in the prior art, owing to be not that every kind of target substance all is fit to separate with high speed adverse current chromatogram, therefore select suitable, effective means of purification to need those skilled in the art to pay performing creative labour;
4, owing to generally be different at the different elution systems that target substance adopted, the difference of elution system will directly influence the effect of wash-out, the inventor is surprised to find that, adopt the elution system (gradient system that comprises macroporous resin, polymeric amide that the present invention limited, the elution system of HSCCC) can obtain best elute effect, make under equal conditions, the content of potentilla flavone is the highest in the product, and selection can obtain so that effect is that those skilled in the art are unforeseen like this;
5, for other conditions and parameter in the production technique, for example chromatographic column model, solvent load, flow velocity, applied sample amount, collection time etc., the contriver investigates by various single factors and has carried out meticulous selection, has obtained best productive rate thus, and this also is that those skilled in the art are unpredictable.
Embodiment
Embodiment 1.
Take from right drying, Chinese medicine tormentic 25kg is produced in Ji County, Tianjin of pulverizing, adds 200kg 60% alcohol immersion and spends the night, and heating and refluxing extraction 1.5 hours is filtered, and with 150kg 60% alcohol reflux 1.5 hours, filters then; And then with 150kg60% alcohol reflux 1.5 hours, merge No. three times extracting solution, concentrate medicinal extract 4.6kg, the HPLC method is measured potentilla flavone content 4.050 ‰.
Above extract is with 46kg 30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min gets supernatant liquor, separate with 20-60 order AB-8 macroporous adsorptive resins, the last sample of 10BV, the absorption flow velocity is 2BV/h, then successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h, collect 60% ethanol eluate, being evaporated to does not have the alcohol flavor, obtains a suspension A (the tormentic flavones content is 3.2% in this extract).With gained suspension A with equal-volume 30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min, getting supernatant liquor separates with 80-100 order polymeric amide chromatographic column, the last sample of 10BV, the absorption flow velocity is 2BV/h, then successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h, collect 60% ethanol eluate, being evaporated to does not have the alcohol flavor, obtains a suspension B (content of potentilla flavone is 12.5% in this mixture).
Suspension B is centrifugal, the centrifugal 10min of 3000r/min, precipitation after centrifugal is with preparation high-speed counter-current chromatograph (TBE-300A) purifying: solvent systems (volume ratio): methylene dichloride: methyl alcohol: water=5: 4: 2 below is stationary phase mutually, on be moving phase mutually, flow velocity 2ml/min, counter-rotating separates, rotating speed 800rpm, 25 ℃ of temperature, detect wavelength 254nm, each sample size 0.5g.Collect flow point: 13min begins to collect behind the beginning appearance time, collects the wash-out flow point of 20min time period.The collection flow point is removed organic solvent through concentrating under reduced pressure and is got an aqueous suspension, and the centrifugal yellow mercury oxide that gets with a small amount of 30% ethanolic soln rinse 2 times, promptly gets potentilla flavone elaboration 3.12g (it is 94.5% that HPLC measures potentilla flavone content, the peak area normalization method).
Embodiment 2.
Take from right drying, Chinese medicine tormentic 30kg is produced in Ji County, Tianjin of pulverizing, adds the 240kg60% alcohol immersion and spends the night, and heating and refluxing extraction 1.5 hours is filtered, and with 180kg60% alcohol reflux 1.5 hours, filters then; And then with 180kg 60% alcohol reflux 1.5 hours, merge No. three times extracting solution, concentrate medicinal extract 5.4kg, wherein potentilla flavone content 4.022 ‰.With the 54kg30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min gets supernatant liquor, separate with 20-60 order AB-8 macroporous adsorptive resins, the last sample of 10BV, the absorption flow velocity is 2BV/h, then successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h, collect 60% ethanol eluate, being evaporated to does not have the alcohol flavor, obtains an aqueous suspension A (the tormentic flavones content is 3.4% in this extract).With the gained suspension with equal-volume 30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min gets supernatant liquor and separates the last sample of 10BV with the polymeric amide chromatographic column, the absorption flow velocity is 2BV/h, successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h collects 60% ethanol eluate then, being evaporated to does not have the alcohol flavor, obtains a suspension B (content of potentilla flavone is 12.1% in this separated portion).
Suspension B is centrifugal, the centrifugal 10min of 3000r/min, precipitation after centrifugal is with preparation high-speed counter-current chromatograph (TBE-300A) purifying: solvent systems (volume ratio): methylene dichloride: methyl alcohol: water=5: 4: 2 below is stationary phase mutually, on be moving phase mutually, flow velocity 2ml/min, counter-rotating separates, rotating speed 800rpm, 25 ℃ of temperature, detect wavelength 254nm, each sample size 0.5g.Collect flow point: 13min begins to collect behind the beginning appearance time, collects the wash-out flow point of 20min time period.The collection flow point is removed organic solvent through concentrating under reduced pressure and is got an aqueous suspension, and the centrifugal yellow mercury oxide that gets with a small amount of 30% ethanolic soln rinse 2 times, promptly gets potentilla flavone elaboration 3.68g (it is 92.8% that HPLC measures potentilla flavone content, the peak area normalization method).
Embodiment 3.
Take from right drying, Chinese medicine tormentic 35kg is produced in Ji County, Tianjin of pulverizing, adds the 280kg60% alcohol immersion and spends the night, and heating and refluxing extraction 1.5 hours is filtered, and with 210kg60% alcohol reflux 1.5 hours, filters then; And then with 210kg 60% alcohol reflux 1.5 hours, merge No. three times extracting solution, concentrate fluid extract 6.1kg, wherein potentilla flavone content 4.502 ‰.With the 61kg30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min gets supernatant liquor, separate with the AB-8 macroporous adsorptive resins, the last sample of 10BV, the absorption flow velocity is 2BV/h, then successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h, collect 60% ethanol eluate, being evaporated to does not have alcohol flavor, the suspension A that obtains (the tormentic flavones content is 3.6% in this extract).With suspension A with equal-volume 30% dissolve with ethanol, fully stir, the centrifugal 10min of 3000r/min gets supernatant liquor and separates the last sample of 10BV with the polymeric amide chromatographic column, the absorption flow velocity is 2BV/h, successively with 3BV mass concentration 30% ethanol, 5BV mass concentration 60% ethanol, 4BV mass concentration 95% ethanol elution, elution flow rate 2BV/h collects 60% ethanol eluate then, concentrating under reduced pressure obtains suspension B (content of potentilla flavone reaches 12.7% in this separated portion).
Suspension B is centrifugal, the centrifugal 10min of 3000r/min, precipitation after centrifugal is with preparation high-speed counter-current chromatograph (TBE-300A) purifying: solvent systems (volume ratio): methylene dichloride: methyl alcohol: water=5: 4: 2 below is stationary phase mutually, on be moving phase mutually, flow velocity 2ml/min, counter-rotating separates, rotating speed 800rpm, 25 ℃ of temperature, detect wavelength 254nm, each sample size 0.5g.Collect flow point: 13min begins to collect behind the beginning appearance time, collects the wash-out flow point of 20min time period.The collection flow point is removed organic solvent through concentrating under reduced pressure and is got an aqueous suspension, and the centrifugal yellow mercury oxide that gets with a small amount of 30% ethanolic soln rinse 2 times, promptly gets potentilla flavone elaboration 4.46g (it is 93.8% that HPLC measures potentilla flavone content, the peak area normalization method).
Appendix: the physicochemical data of embodiment 1-3 the finished product-potentilla flavone and pharmacology evaluation
1. the physicochemical data of potentilla flavone (formula 1):
Yellow unformed powder.The intensive uv-absorbing is arranged under the UV356.ESI-MS?m/z：593[M-H]
-(C
30H
25O
13)。HRESI-MS:m/z 617.1284[M+Na]+calcd.617.1271, C
30H
26O
13Na; M/z 593.1204[M-H]
-, calcd.593.1295, C
30H
25O
13. determine that therefore this compound molecule formula is C
30H
26O
13
1H-NMR(CD
3OD)δ：6.14(1H，br?s，6-H)，6.31(1H，br?s，8-H)，7.97(2H，d，J＝8.7Hz，H-2`，6`)，6.82(2H，d，J＝8.7Hz，H-3`，5`)，7.300(2H，d，J＝8.8Hz，H-5```，9```)，6.79(2H，d，J＝8.8Hz，H-6```，8```)，6.06(1H，d，J＝15.9Hz，H-2```)，7.40(1H，d，J＝15.9Hz，H-3```)，5.23(1H，d，J＝7.8Hz，H-1``)，4.18～4.30(2H，m，H-6``)，3.50(2H，m)，3.32(1H，m，H-4``)。
13C-NMR (CD
3OD) aglycon: 159.4,135.4,179.5,163.1,100.1,166.0,95.0,158.2,105.7,122.9,132.3,116.2,161.6,116.2,132.2; Glucosyl group: 104.2,78.2,75.8,71.9,75.9,64.4; Trans to the hydroxyl cinnamyl: 168.9,115.0,146.6,127.3,131.2,116.9,161.2,116.9,131.2.Above spectroscopic data and document [Sachiko T.; Kyoko T.; Maki A.; et al; Isolation of Cytochrome P450 Inhibitors fromStrawberry Fruit; Fragaria ananassa; J.Nat.Prod.2004; 67:1839～1841.] report kaempferol-3-O-β-D-(6-O-trans-is to the hydroxyl cinnamyl) glycoside spectral data unanimity; therefore, identify that this compd E formula is kaempferol-3-O-β-D-(6-O-trans-is to the hydroxyl cinnamyl) glycoside (kaempferol-3-O-β-D-(6-O-trans-p-coumaroyl) glucopyranoside).
2. the anti-diabetic pharmacology of potentilla flavone is estimated
2.1 the hypoglycemic effective constituent dose-effect relationship of tormentic is investigated
2.1.1 grouping and administration
Choose normal mouse, set up tetraoxypyrimidine hyperglycemia diabetic mice model, be divided into 6 groups at random by the even principle of blood glucose value.The potentilla flavone maximum dose level (1.6mg/kg), high dosage (0.8mg/kg), middle dosage (0.4mg/kg), the low dose group (0.1mg/kg) that are divided into model control group, N1,N1-Dimethylbiguanide positive controls (125mg/kg) and formula I of the present invention.Other is provided with normal control group and normal administration group, and the normal control group gives distilled water (10ml/kg), and normal administration group gives potentilla flavone by 0.4mg/kg dosage, 10 every group.Below respectively organize and irritate stomach every day 1 time, continuous irrigation stomach 15 days.Respectively organize rat during the medication treatment and all raise with normal diet ad lib, drinking-water every day.
2.1.2 potentilla flavone is to the influence of tissue of experimental diabetic mice blood sugar
Table 1 potentilla flavone is to the influence of tissue of experimental diabetic mice fasting plasma glucose (x ± s)
*With with time model group p＜0.05 relatively,
*With with time model group p＜0.01 relatively,
△ with trans low dose group of time p＜0.05 relatively
As can be seen, each group of diabetic mice after potentilla flavone treatment 7 days, only trans potentilla flavone 1.6mg/kg dosage group and model group relatively, fasting plasma glucose has significant difference (P＜0.05), all the other each difference of organizing that there are no significant.Each group of diabetic mice after 15 days, is compared all significantly reduction (P＜0.05, P＜0.01) through the potentilla flavone treatment with fasting plasma glucose before the administration.After the administration, each dosage group of trans potentilla flavone and model group relatively fasting plasma glucose also significantly reduce (P＜0.05, P＜0.01), and each group of trans potentilla flavone presents certain dose-effect relationship and time-effect relationship on hypoglycemic activity.Normal mouse administration group illustrates that trans potentilla flavone does not exert an influence to the glucose level of normal mouse with the blank group and relatively there are no significant before the administration on the same group difference.
2.1.3 potentilla flavone is to the influence of tissue of experimental diabetic mice blood fat
As can be seen from Table 2, after the administration, the total cholesterol concentration of 4 dosage groups of trans potentilla flavone and cis group and triglyceride concentration and model group more all have significant difference (P＜0.01, P＜0.05).The result of trans four various dose groups shows that the average that increases triglyceride level and total cholesterol level along with dosage presents the trend that progressively reduces.And after the administration four dosage groups of trans potentilla flavone and cis group with two indexs of positive controls relatively there are no significant difference, illustrate that to a certain extent potentilla flavone can reach the effect of Walaphage aspect triglyceride reducing and total cholesterol concentration.And two indexs respectively with blank group relatively there are no significant difference, the illustration effect is remarkable, it is unusual partly to correct the lipid metabolism that diabetes cause.Make the triglyceride level of diabetic mice and the level that total cholesterol concentration reaches normal mouse.Normal mouse administration group illustrates that trans potentilla flavone does not exert an influence to the total cholesterol and the triglyceride levels of normal mouse with the blank group and relatively there are no significant before the administration on the same group difference.
Table 2 potentilla flavone is to the influence of tissue of experimental diabetic mice triglyceride level and total cholesterol (x ± s)
*Compare p＜0.05 with model group,
*Compare p＜0.01 with model group,
★Compare p＜0.05 with the blank group,
★ ★Compare p＜0.01 with the blank group
Claims (1)
1. tormentic effective constituent chromocor compound process for extracting, separating and purifying, described tormentic anti-diabetic chemical ingredients, its structural formula is as follows:
The process for extracting, separating and purifying of above-mentioned potentilla flavone is characterized in that comprising following process:
(1) tormentic is pulverized, mass concentration 60% alcohol immersion with 8 times of the quality of tormentic is spent the night, heating and refluxing extraction 1.5 hours, filtering filter residue, mass concentration 60% alcohol heating reflux with 6 times of the quality of tormentic extracted 1.5 hours again, filtering filter residue, and mass concentration 60% alcohol heating reflux with 6 times of the quality of tormentic extracted 1.5 hours again, merge the filtrate of three extractions, reduction vaporization is concentrated into and soaks paste;
(2) paste that step (1) is obtained is measured mass concentration 30% dissolve with ethanol with 10 times, the centrifugal 10min of 3000r/min separates last sample volume 10BV (10 times of column volumes) with 20-60 purpose AB-8 macroporous adsorptive resins, the absorption flow velocity is 2 BV/h, then with 3 BV30% ethanol, 5 BV60% ethanol, 4 BV95% ethanol are wash-out successively, elution flow rate 2 BV/h, collect 60% ethanol eluate, being evaporated to does not have the alcohol flavor, gets a suspension A;
(3) the suspension A that step (2) is obtained is with equal-volume 30% dissolve with ethanol, the centrifugal 10min of 3000r/min separates last sample volume 10BV with 80-100 order polymeric amide chromatographic column, the absorption flow velocity is 2BV/h, then with 3BV30% ethanol, 5BV60% ethanol, 4BV95% ethanol is wash-out successively, with elution flow rate 2BV/h, collect 60% ethanol eluate, being evaporated to does not have the alcohol flavor, gets a suspension B;
(4) suspension B that step (3) is obtained is centrifugal, the centrifugal 10min of 3000r/min, precipitation after centrifugal is to prepare the high-speed counter-current chromatograph purifying: solvent systems (volume ratio): methylene dichloride: methyl alcohol: water=5: 4: 2, below be stationary phase mutually, on be moving phase mutually, flow velocity 2ml/min, counter-rotating separates, rotating speed 800rpm, 25 ℃ of temperature detect wavelength 254nm.Collect flow point: 13min begins to collect behind the beginning appearance time, collects the wash-out flow point of 20min time period.The collection flow point is removed organic solvent through concentrating under reduced pressure and is got an aqueous suspension, and the centrifugal yellow mercury oxide that obtains with a small amount of 30% ethanolic soln rinse 2 times, promptly gets the potentilla flavone elaboration.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103209971A (en) * | 2010-11-12 | 2013-07-17 | 天津医科大学 | Flavone derivatives and their preparative method and medical use |
| CN104861020A (en) * | 2015-06-14 | 2015-08-26 | 海南医学院 | Phenylpropanoid glycoside compound, and extraction method and application thereof |
| CN104945455A (en) * | 2014-03-28 | 2015-09-30 | 中国医学科学院药物研究所 | Coumarin glycoside compound, and preparation method, pharmaceutical composition and application preparation thereof |
| CN107459543A (en) * | 2017-08-28 | 2017-12-12 | 天津科技大学 | A kind of preparation method of astragalin derivative |
| CN111548386A (en) * | 2020-05-28 | 2020-08-18 | 广西中医药大学第一附属医院 | Method for extracting tormentic acid and application thereof |
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2009
- 2009-12-17 CN CN200910244861A patent/CN101735292A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103209971A (en) * | 2010-11-12 | 2013-07-17 | 天津医科大学 | Flavone derivatives and their preparative method and medical use |
| CN103209971B (en) * | 2010-11-12 | 2014-08-20 | 天津医科大学 | Flavone derivatives and their preparative method and medical use |
| CN104945455A (en) * | 2014-03-28 | 2015-09-30 | 中国医学科学院药物研究所 | Coumarin glycoside compound, and preparation method, pharmaceutical composition and application preparation thereof |
| CN104945455B (en) * | 2014-03-28 | 2019-01-01 | 中国医学科学院药物研究所 | Tonka bean camphor glycosides compounds, its preparation method and pharmaceutical composition and purposes |
| CN104861020A (en) * | 2015-06-14 | 2015-08-26 | 海南医学院 | Phenylpropanoid glycoside compound, and extraction method and application thereof |
| CN104861020B (en) * | 2015-06-14 | 2017-05-17 | 海南医学院 | Phenylpropanoid glycoside compound, and extraction method and application thereof |
| CN107459543A (en) * | 2017-08-28 | 2017-12-12 | 天津科技大学 | A kind of preparation method of astragalin derivative |
| CN107459543B (en) * | 2017-08-28 | 2020-07-28 | 天津科技大学 | Preparation method of astragalin derivative |
| CN111548386A (en) * | 2020-05-28 | 2020-08-18 | 广西中医药大学第一附属医院 | Method for extracting tormentic acid and application thereof |
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Application publication date: 20100616 |