CN1312170C - Technique of preparing extract product of Radde Anemone Rhizome extract, and its application in preparing medication of treating cancer - Google Patents
Technique of preparing extract product of Radde Anemone Rhizome extract, and its application in preparing medication of treating cancer Download PDFInfo
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Abstract
The present invention discloses a preparation technology of a biapiculate extract product. Biapiculate medicinal materials are used as raw materials, and the treatment, such as alkali extraction, acid neutralization, condensation, alcohol deposition, etc., is carried out to the biapiculate medicinal materials which are further purified to obtain the biapiculate extract product. The purity and the yield of anemonin A which is a constituent with strong anti-cancer activity in the obtained biapiculate extract product are both high, the purity can reach more than 50 percent, and the yield is calculated in a mode that the anemonin A obtained by 1 kilogram of medicinal materials can reach more than 10 grams. The operation of the present invention is simple, and the present invention is suitable for commercial process. The present invention also discloses the application of the extract product obtained by the preparation technology in preparing medicines for treating cancer, the biapiculate extract product containing treatment effective amount and a carrier acceptable by pharmacology are made into certain dosage form medicines which have the advantages of strong antitumous effects, little daily amount, convenient taking, etc.
Description
Technical field
The present invention relates to a kind of extracting method of effective ingredient in Chinese, more particularly, the present invention relates to a kind of method of from Chinese medicine is pointed at both ends, extracting efficient part.The invention still further relates to by the application of the resulting Radde anemone rhizome extract of this method in the medicine of preparation treatment cancer.
Background technology
Pointed at both ends is the dry rhizome of Ranunculaceae thimbleweed anemone raddeana Regel (Anemone raddeana Regel), also name Radde Anemone Rhizome, radix aconiti agrestis beak, the 133rd page of record effect with " wind-damp dispelling; it is too fat to move to disappear " pointed at both ends of the Pharmacopoeia of the People's Republic of China (version was an one in 2000) text, be used for the treatment of " arthralgia due to wind-cold-dampness pathogen BI syndrome; spasm of the limbs, arthralgia, too fat to move festering " etc.
Modern pharmacy studies show that, pointed at both ends also have antitumor, anti-inflammatory, analgesia, an effect such as anticonvulsion.That one literary composition report thimbleweed such as " time precious traditional Chinese medical science traditional Chinese medicines " (1999) the 10th volume the 1st phase Liu Yonghong, Liu Dayou " the medicinal new developments of thimbleweed " has is antitumor, anti-inflammatory, analgesia, effect such as anticonvulsion, especially antitumour activity is remarkable, its total saponin, especially saponin D are the generation that anemonin A (Raddeanin A) can suppress cancer." the plain A of anemone raddeana Regel is to cancer cells DNA, RNA to roll up " Acta Pharmacologica Sinica " (in September, 1985) the 6th such as the 3rd phase Liu's Lik-Sang, Wu Fenge etc., the influence of protein and plasma cAMP content " anemonin A of a literary composition report significant quantity can significantly suppress mouse S180 and ascites hepatoma cells DNA external, RNA and proteinic synthetic, thus favourable to anticancer." research of anemonin A and Altay black false hellebore total alkali mutagenesis " literary compositions such as " Lanzhou University's journal " (natural science edition) 21 (biology is collected periodical) in December, 1985 Wang Xiao tawny daylily, Liu Qin have also been measured the effect that two kind of plant natural product anemonin As, Altay black false hellebore total alkali bring out human lymphocyte SCE by SCD technology (younger sister's sister-in-law chromatid differential stain method), and the relation between dosage and the effect and comparison the two bring out the size of SCE ability, thereby show that anemonin A may not be mutagenic compound and may be comparatively ideal anticarcinogen." chemical journal " (1984) the 42nd volume the 3rd phase Wu Feng blades of a sword " Chinese medicine Radde Anemone Rhizome The Chemical Constituents " literary composition report anemonin A is the main active ingredient that has antitumous effect in pointed at both ends.
Chinese patent publication number CN1312253A, open day September 12 calendar year 2001, the name of innovation and creation is called the preparation technology and the purposes of Radde anemone rhizome extract, this application case discloses a kind of preparation technology and purposes of Radde anemone rhizome extract, from pointed at both ends and stem, leaf, flower, fruit, extract total saponin and saponin monomer D with pharmaceutical use, and make medicine, be used for the treatment of malignancy disease.The preparation technology of its total saponin mainly may further comprise the steps: use the 50-70% extraction using alcohol, or extract with decoction alcohol precipitation method, handle through macroporous adsorbent resin then, obtain total saponin pointed at both ends.Its weak point is that to extract the content of anemonin A in the total saponin obtain lower, and the yield of anemonin A is also lower.
Chinese patent publication number CN1342655A, open day on April 3rd, 2002, the name of innovation and creation is called the conversion of saponin in the Radde Anemone Rhizome and the extraction process of total saponin and anemonin A, and this application case discloses and a kind ofly adopted basic hydrolysis to make to contain the extraction process that the saponin of sugar more than three is converted into anemonin A and total saponin and anemonin A in the total saponin of Radde Anemone Rhizome.The preparation technology of its total saponin mainly may further comprise the steps: boiled 1-8 hour or mix the back thoroughly with alkali aqueous solution and steamed 1-5 hour with high-temperature vapour with alkali aqueous solution, make and contain the saponin of sugar more than three in the total saponin of Radde Anemone Rhizome and be converted into anemonin A, again with the acid neutralization, extract with 50-100% alcohol or ethyl acetate backflow, filter evaporate to dryness, get total saponin.Its weak point is under the described condition of invention, when containing the saponin hydrolysis of sugar more than three in the total saponin of Radde Anemone Rhizome and be converted into anemonin A, anemonin A is partial hydrolysis simultaneously also, thereby, in total saponin that extraction obtains, the content of anemonin A and yield are all not high.
Summary of the invention
The objective of the invention is, at the content and all lower shortcoming of yield thereof of the stronger composition anemonin A of antitumour activity in the Radde anemone rhizome extract in the prior art, provide the preparation technology and the application of extract in the medicine of preparation treatment cancer thereof of all higher Radde anemone rhizome extract of a kind of yield of anemonin A and purity.
The preparation technology of Radde anemone rhizome extract among the present invention comprises following processing step:
(1) get pointed at both ends or pulverize after pointed at both ends, add alkali lye, extract, extracting solution.Wherein, after adding alkali lye, can soak earlier again and extract, preferably soak after 6-48 hour and extract again; Extracting method can be the traditional Chinese medicine extraction method of any conventional, preferred percolation, decocting method or reflux extraction, preferably percolation; The preferred 4-40 of amount that adds alkali lye doubly; Alkali lye can be alkaline aqueous solution or alkaline alcohol solution, preferred alkaline aqueous solution: the preferred 9-13 of the pH value of alkali lye.
(2) be 4-8 with sour neutralising extract to pH value, be preferably 6-7.5; It is 0.95-1.30 that extracting solution after the neutralization is concentrated into relative density, and adding ethanol to concentration is 50-95%, leaves standstill certain hour, preferably more than 6 hours; Get supernatant liquid and reclaim ethanol, get a suspended matter.
(3) the centrifugal or filtration with this suspended matter must precipitate.
(4) this precipitation is dissolved in 60-100% ethanol, adding water adjusting alcohol concn is 5-50%, preferred 20-40%, and precipitation is got in centrifugal or filtration, promptly obtains Radde anemone rhizome extract.
If necessary, but the operation of gained said extracted thing repeating step (4) with further separation and purification, obtains the higher extract of anemonin A content.
The application of the Radde anemone rhizome extract that obtains by above preparation technology among the present invention in the medicine of preparation treatment cancer, its solution is, what will contain the treatment significant quantity makes the medicine of any formulation, preferred tablet, granule, capsule or injection injection etc. according to the resulting Radde anemone rhizome extract of above-described preparation technology and pharmaceutically acceptable carrier.Be 0.1-5 milligram/day kilogram wherein, be equivalent to raw medicinal herbs 60-300 milligram/day kilogram according to the treatment significant quantity of the resulting Radde anemone rhizome extract of above-described preparation technology.
An important feature of the present invention is, purity and yield by the composition anemonin A that antitumour activity is stronger in the resulting Radde anemone rhizome extract of the above preparation technology are all higher, purity can reach more than 50%, and yield obtains anemonin A and can reach more than 10 grams in 1 kilogram of medicinal material; And this technological operation is simple, is applicable to suitability for industrialized production.
Another important feature of the present invention is, by containing the medicine that the above Radde anemone rhizome extract for the treatment of significant quantity and pharmaceutically acceptable carrier are made, has advantages such as stronger, the daily dosage of antitumous effect is little, easy administration.
Embodiment
The invention will be further described below in conjunction with embodiment.
Get pointed at both ends or pulverize after pointed at both ends, add alkali lye and extract, extracting solution; With sour neutralising extract to pH value is 4-8, and it is 0.95-1.30 that the extracting solution after the neutralization is concentrated into relative density, and adding ethanol to concentration is 50-95%, leaves standstill, and gets supernatant liquid and reclaims ethanol, gets a suspended matter; Centrifugal or filtration must precipitate with this suspended matter; This precipitation is dissolved in 60-100% ethanol, and adding water adjusting alcohol concn is 5-50%, and precipitation is got in centrifugal or filtration, obtains Radde anemone rhizome extract.
Above-mentioned Radde anemone rhizome extract, further separation and purification is removed impurity, to obtain the higher product of anemonin A content in case of necessity.
Be the Comparative Examples and the embodiment of preparation Radde anemone rhizome extract below.Can recognize the influence of different technology conditions to anemonin A purity and yield by Comparative Examples and embodiment, wherein yield is to obtain what (gram/kilogram medicinal materials) of anemonin A from the per kilogram medicinal material.
Comparative Examples 1:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, the NaOH aqueous solution that adds 10 kilogram 1%, little boiling 2 hours, neutralize with 50% ethanol solution of sulfuric acid again, regulate pH value to 8, the sample concentration after the neutralization is to half-dried, with evaporate to dryness again after an amount of 95% alcohol reflux stripping, residue is used 95% dissolve with ethanol again, filter the back evaporate to dryness, dissolve once more with 95% ethanol afterwards, filtrate reclaims solvent, vacuum-drying, get total saponin 52 grams, wherein anemonin A content is 7.2%, and the anemonin A yield is 3.74 gram/kilogram medicinal materials.
Comparative Examples 2:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, 70% ethanolic soln that adds 12 kilograms, refluxing extraction 3 times each 2 hours, is filtered extracting solution and is merged, reclaim under reduced pressure is to there not being the alcohol flavor, dry back adds 70% dissolve with ethanol solution, and last D101 macroporous adsorptive resins washes with water after colourless, ethanolic soln with 70% is eluted to till the no saponin reaction, with 1% activated carbon decolorizing 30 minutes, filtered while hot reclaimed ethanol to there not being the alcohol flavor with elutriant, dry, get tawny powder 33.9 grams, wherein anemonin A content is 18.75%, and the anemonin A yield is 6.36 gram/kilogram medicinal materials.
Comparative Examples 3:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 15 kg of water, decoct 3 times, each 2 hours, filter, collecting decoction, being concentrated into relative density is 1.20, adds ethanol, make ethanol content reach 70%, left standstill 24 hours, filter, reclaim ethanol to 2050ml, add ethanol and make pure content reach 90%, left standstill 24 hours, filter, filtrate recycling ethanol is to there not being the alcohol flavor, dry back adds 70% dissolve with ethanol solution, and last D101 macroporous adsorptive resins washes with water after colourless, ethanolic soln with 70% is eluted to till the no saponin reaction, with 1% activated carbon decolorizing 30 minutes, filtered while hot reclaimed ethanol to there not being the alcohol flavor with elutriant, dry, get tawny powder 23.7 grams, wherein anemonin A content is 13.23%, and the anemonin A yield is 3.14 gram/kilogram medicinal materials.
Comparative Examples 4:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, the ethanolic soln of adding 70%, diacolation is to there not being the saponin reaction, percolate is reclaimed ethanol to there not being the alcohol flavor, dry back adds 70% dissolve with ethanol solution, and last D101 macroporous adsorptive resins washes with water after colourless, ethanolic soln with 70% is eluted to till the no saponin reaction, with 1% activated carbon decolorizing 30 minutes, filtered while hot reclaimed ethanol to there not being the alcohol flavor with elutriant, dry, get tawny powder 29.2 grams, wherein anemonin A content is 20.15%, and the anemonin A yield is 5.88 gram/kilogram medicinal materials.
Embodiment 1:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 2 kilograms of pH values and be 13.6 NaOH solution, soaked 4 hours, the beginning diacolation, diacolation speed is 2ml/min, the diacolation time is 25 hours.Merge percolate, transfer pH value to 6.5, be evaporated to relative density 0.95 with the hydrochloric acid of 2M, adding ethanol to determining alcohol is 80%, places 2 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, filters, precipitation is dissolved in 90% ethanol, adds water to determining alcohol 5%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 16 grams, wherein anemonin A content is 65.04%, and the anemonin A yield is 10.41 gram/kilogram medicinal materials.
Embodiment 2:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 4 kilograms of pH values and be 13 KOH solution, soaked 24 hours, the beginning diacolation, diacolation speed is 5ml/min, the diacolation time is 40 hours.Merge percolate, transfer pH value to 7.0, be evaporated to relative density 1.30 with the sulfuric acid of 1M, adding ethanol to determining alcohol is 70%, places 16 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, and is centrifugal, precipitation is dissolved in 60%7 alcohol, adds water to determining alcohol 30%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 22 grams, wherein anemonin A content is 68.08%, and the anemonin A yield is 14.97 gram/kilogram medicinal materials.
Embodiment 3:
Getting 1 kilogram pointed at both ends, be ground into 20 order meal, is that 12.69 NaOH solution carries out diacolation with pH value, and diacolation speed is 2ml/min, and the diacolation time is 70 hours.Merge percolate, transfer pH value to 5.0, be evaporated to relative density 1.02 with the sulfuric acid of 1M, adding ethanol to determining alcohol is 85%, places 8 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, and is centrifugal, precipitation is dissolved in 70% ethanol, adds water to determining alcohol 40%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 19 grams, wherein anemonin A content is 71.12%, and the anemonin A yield is 13.51 gram/kilogram medicinal materials.
Embodiment 4:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 4 kilograms of pH values and be 11 Ca (OH)
2Solution soaked 8 hours, the beginning diacolation, and diacolation speed is 20ml/min, the diacolation time is 15 hours.Merge percolate, transfer pH value to 4.5, be evaporated to relative density 1.15 with the sulfuric acid of 1M, adding ethanol to determining alcohol is 75%, places 24 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, and is centrifugal, precipitation is dissolved in 60% ethanol, adds water to determining alcohol 10%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 19 grams, wherein anemonin A content is 72.35%, and the anemonin A yield is 13.75 gram/kilogram medicinal materials.
Embodiment 5:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 4 kilograms of pH values and be 10 ammonia soln, soaked 3 hours, the beginning diacolation, diacolation speed is 2ml/min, the diacolation time is 90 hours.Merge percolate, transfer pH value to 6.0, be evaporated to relative density 1.20 with the hydrochloric acid of 2M, adding ethanol to determining alcohol is 80%, places 16 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, and is centrifugal, precipitation is dissolved in 80% ethanol, adds water to determining alcohol 20%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 25 grams, wherein anemonin A content is 70.23%, and the anemonin A yield is 17.56 gram/kilogram medicinal materials.
Embodiment 6:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 6 kilograms of pH values and be 9 NaOH solution, soaked 48 hours, the beginning diacolation, diacolation speed is 5ml/min, the diacolation time is 50 hours.Merge percolate, transfer pH value to 7.0, be evaporated to relative density 1.06 with the sulfuric acid of 1M, adding ethanol to determining alcohol is 85%, places 4 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, and is centrifugal, precipitation is dissolved in 95% ethanol, adds water to determining alcohol 50%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 18 grams, wherein anemonin A content is 66.21%, and the anemonin A yield is 11.92 gram/kilogram medicinal materials.
Embodiment 7:
Get 1 kilogram pointed at both ends, be ground into 20 order meal, add 6 kilograms of pH values and be 8.6 NaOH solution, soaked 48 hours, the beginning diacolation, diacolation speed is 2ml/min, the diacolation time is 120 hours.Merge percolate, transfer pH value to 6.0, be evaporated to relative density 1.25 with the sulfuric acid of 1M, adding ethanol to determining alcohol is 80%, places 24 hours, draws supernatant liquor, decompression recycling ethanol has precipitation to separate out to there not being the alcohol flavor this moment, and is centrifugal, precipitation is dissolved in 85% ethanol, adds water to determining alcohol 40%, leave standstill a moment, centrifugal, get faint yellow precipitation, dry back weighs 18 grams, wherein anemonin A content is 70.20%, and the anemonin A yield is 12.64 gram/kilogram medicinal materials.
What will contain the treatment significant quantity makes the medicine of any formulation according to the resulting Radde anemone rhizome extract of the above preparation technology and pharmaceutically acceptable carrier.
Be the embodiment of the application of Radde anemone rhizome extract in the medicine of preparation treatment cancer below.
Embodiment 1: make tablet
Radde anemone rhizome extract 210g and starch 80g are mixed, and the starch slurry with 10% is granulated, and dries in 65 ℃ baking oven, adds the 3g Magnesium Stearate, mixes, and compressing tablet is made 1000, and every contains 0.3g.
Embodiment 2: make granule
With Radde anemone rhizome extract 2000g and starch 200g, dextrin 800g, mix, granulate in right amount with 80% ethanol, in 80 ℃ of dryings, be distributed into 1000 bags, every 3g.
Embodiment 3: make capsule
With Radde anemone rhizome extract 200g and W-Gum 97g mixing and stirring, add the moistening granulation of an amount of 95% ethanol, drying adds the 3g Magnesium Stearate, mixes, and in incapsulating, makes 1000 of capsules, and every contains content 0.3g.
Embodiment 4: make the injection injection
It is among 12 the NaOH solution 600ml that Radde anemone rhizome extract 0.6g is dissolved in pH value, and transferring pH value with the salt of 0.05mol/l is 7.5, use the millipore filtration suction filtration, adds and injects water to 1000ml, and the ampoule that specification is 2ml is gone in embedding under aseptic condition.
Claims (14)
1. the preparation technology of a Radde anemone rhizome extract comprises following processing step:
(1) get pointed at both ends or pulverize after pointed at both ends, add alkali lye and extract, extracting solution;
(2) be 4-8 with sour neutralising extract to pH value, it is 0.95-1.30 that the extracting solution after the neutralization is concentrated into relative density, and adding ethanol to determining alcohol is 50-95%, leaves standstill, and gets supernatant liquid and reclaims ethanol, gets a suspended matter;
(3) the centrifugal or filtration with this suspended matter must precipitate;
(4) this precipitation is dissolved in 60-100% ethanol, adding water adjusting alcohol concn is 5-50%, and precipitation is got in centrifugal or filtration, obtains Radde anemone rhizome extract.
2. preparation technology according to claim 1 wherein adds in the step (1) to soak earlier behind the alkali lye again and extracts.
3. preparation technology according to claim 1 wherein adds in the step (1) to soak earlier behind the alkali lye and extracted in 6-48 hour again.
4. preparation technology according to claim 1, wherein extracting method is percolation, decocting method or reflux extraction in the step (1).
5. preparation technology according to claim 1, wherein extracting method is a percolation in the step (1).
6. preparation technology according to claim 1, wherein alkali lye is alkaline aqueous solution or alkaline alcohol solution in the step (1).
7. preparation technology according to claim 1, wherein alkali lye is alkaline aqueous solution in the step (1).
8. preparation technology according to claim 1, the amount that wherein adds alkali lye in the step (1) are 4-40 times.
9. preparation technology according to claim 1, wherein the alkali lye pH value is 9-13 in the step (1).
10. preparation technology according to claim 1, wherein step (2) is middle is 6-7.5 with sour neutralising extract to pH value.
11. preparation technology according to claim 1, wherein time of repose is more than 6 hours in the step (2).
12. preparation technology according to claim 1, wherein adding water adjusting alcohol concn in the step (4) is 20-40%.
13. the application of the Radde anemone rhizome extract that preparation technology according to claim 1 obtains in the medicine of preparation treatment cancer will contain the medicine that the resulting Radde anemone rhizome extract of preparation technology according to claim 1 for the treatment of significant quantity and pharmaceutically acceptable carrier are made any formulation.
14. the application of Radde anemone rhizome extract according to claim 13 in the medicine of preparation treatment cancer, wherein said formulation is tablet, granule, capsule or injection injection.
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| CN1307192C (en) * | 2005-05-24 | 2007-03-28 | 中国人民解放军第二军医大学 | Process of preparing high-purity jasminodin with Cape jasmine fruit |
| CN101062045B (en) * | 2006-04-29 | 2011-06-22 | 华北制药集团新药研究开发有限责任公司 | Use of triterpene saponin compounds in preparing tumor medicine |
| US8394776B2 (en) | 2007-01-16 | 2013-03-12 | Chengdu Di'ao Jiuhong Pharmaceutical Factor | Use of ursolic acid saponin,oleanolic acid saponin in preparation of increasing leucocytes and/or platelet medicine |
| CN101224215B (en) * | 2007-01-16 | 2010-06-02 | 成都地奥九泓制药厂 | Uses of ursolic acid saponin and oleanolic acid saponin in preparing medicine for increasing white blood cell and/or blood platelet |
| CN101492489B (en) * | 2009-03-06 | 2011-12-07 | 吉林大学 | Method for extracting anemonin A and method of preparing lipid microsphere preparation |
| CN101759706B (en) * | 2009-10-20 | 2012-01-11 | 苏州派腾生物医药科技有限公司 | Method for manufacturing anemonin |
| CN105641078B (en) * | 2014-11-28 | 2019-09-17 | 天津耀宇生物技术有限公司 | From the middle method and its application for extracting active component pointed at both ends |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1312253A (en) * | 2001-01-16 | 2001-09-12 | 长春中医学院 | Radde anemone rhizome extract and its prepn process and use |
| CN1342655A (en) * | 2001-07-19 | 2002-04-03 | 中国科学院成都生物研究所 | Process for conversion of saponin in cyperus tuber and extracting general glycoside and anemonin A |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1312253A (en) * | 2001-01-16 | 2001-09-12 | 长春中医学院 | Radde anemone rhizome extract and its prepn process and use |
| CN1342655A (en) * | 2001-07-19 | 2002-04-03 | 中国科学院成都生物研究所 | Process for conversion of saponin in cyperus tuber and extracting general glycoside and anemonin A |
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