CN105012455A - Pain-easing hemostasis medicine and preparation method and application thereof - Google Patents

Pain-easing hemostasis medicine and preparation method and application thereof Download PDF

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CN105012455A
CN105012455A CN201510508482.6A CN201510508482A CN105012455A CN 105012455 A CN105012455 A CN 105012455A CN 201510508482 A CN201510508482 A CN 201510508482A CN 105012455 A CN105012455 A CN 105012455A
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hemostasis
medicine
pain
easing
preparation
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刘志刚
谢晓林
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Guiyang University
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Guiyang University
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Abstract

The invention relates to pain-easing hemostasis medicine and a preparation method and application thereof and belongs to the technical field of Chinese herb deep processing. With traditional national herbs (Asiatic toddalia roots) being raw materials, the pain-easing hemostasis medicine is obtained through deep processing, and the preparation method and application of the medicine are provided. Extraction is performed on the Asiatic toddalia roots through water and an ethyl alcohol solution, then yellowish-brown particles are obtained through purification and separation, the content of coumarin chemical substances serving as main components is larger than or equal to 25%, the content of alkaloid substances is smaller than 5%, and the balance is mainly starch substances, saccharide substances, tanned substances, terpene substances and sterol substances. The medicine has good pain-easing and hemostatic effects and can be applied to medicine and preparations meeting modern preparation technologies, pharmacodynamics studies show that the pain-easing effect is comparable to that of aspirins, and the hemostatic effect is close to that of Yunnan white drug powder; the technology is simple and convenient to implement, operation is convenient, industrial production can be achieved, and market prospects are good.

Description

A kind of pain easing and hemostasis medicine and its preparation method and application
Technical field
The invention belongs to Chinese medicine field of deep, be specifically related to a kind of a kind of pain easing and hemostasis medicine obtained with the deep processing of ethnic drug Radix Toddaliae Asiaticae and its preparation method and application.
Background technology
Hemostasis is a complicated physiological process, mainly affects by key factors such as blood coagulation, platelet aggregation and vasoconstrictions.When blood vessel injury occurs hemorrhage, the clotting mechanism of self is organized fully to show, thrombokinase increased activity, formation increases, then prothombin is thrombin, thrombin again catalysis fibre proteinogen changes fibrin into, also makes body create antagonism the factor of plasmin solution preocess simultaneously.Every medicine of stopped bleeding that can impel all is called hemorrhage, and hemorrhage mainly by reinforcement intravascular coagulation factor or the solidifying factor of suppression, impels blood coagulation, thus reaches the object of hemostasis.The natural haemostatic medicament used clinically mainly through shortening clotting time, promote local vasoconstriction, strengthen capillary resistance and reduce permeability, suppress fibrinolytic process, increase number of platelets, shorten the thrombin rise time, strengthen effect that the effects such as platelet factor III is active reach hemostasis.Most haemostatic medicament effects conventional are at present single, can not meet clinical needs completely, the existing anastalsis of some natural haemostatic medicaments, can promote blood dissolves, anticoagulant again, and showing clinically stops blooding invigorates blood circulation, peculiar effect of promoting the circulation of blood blood stasis dispelling.
Radix Toddaliae Asiaticae is Rutaceae Radix Toddaliae Asiaticae platymiscium Radix Toddaliae Asiaticae toddalia asiatica(L.) dry root of Lam. or root bark, it is a kind of important national medicine, be widely used in the numerous minority area of south China, recorded by the national medicine ancient books and records such as " Chinese Ben Cao ﹒ Miao Ethnomedicine volume ", " Chinese Ben Cao ﹒ Dai Nationality powder stick ", Miao ethnic group is among the people by various disease conditions such as its water decoction and medicated wine treatment traumatic injury, rheumatic arthritis, venom and traumatic hemorrhage [1].
Although Radix Toddaliae Asiaticae has good pain easing and hemostasis effect [2], but pain easing and hemostasis effect day dosing to be played with reaching about 20-30g, owing to containing more alkaloid toxic component in Radix Toddaliae Asiaticae, if contained chelerythrine is neuromuscular toxin, have strong inhibition effect to heart, excess intake can cause paralysis and death [3]therefore, with Radix Toddaliae Asiaticae pain easing and hemostasis, will take higher dosage just can reach good therapeutic effect on the one hand, on the other hand, dosage can take in more toxin greatly again, for solving this contradiction, necessary by carrying out extracting and developing and purification to Radix Toddaliae Asiaticae, will there is the Coumarins extracts active ingredients of styptic activity out in Radix Toddaliae Asiaticae, to wherein toxic alkaloid component separating go out, develop a kind of dosage few, pain easing and hemostasis active constituent content is high, the medicine that toxic action is low.
[1] herbal editorial board of China of State Administration of Traditional Chinese Medicine. China's book on Chinese herbal medicine, Miao Ethnomedicine is rolled up. Kweiyang, Guizhou Science Press, 2005.
[2] Hao little Yan, Peng Lin, aspidistra, etc. the research of Radix Toddaliae Asiaticae biology total alkali anti-inflammatory and analgesic effect. Journal of Chinese Integrative Medicine, 2004,2 (6): 450-452.
[3] Jia Min is as, Li Xingwei. and China National medicine will is wanted. Beijing, China Medical Science Press, 2005.
Summary of the invention
Object of the present invention, be for Radix Toddaliae Asiaticae pain easing and hemostasis active constituent content low, toxic component content is higher, a large amount of taking orally easily causes poisoning problem, develop a kind of pain easing and hemostasis active constituent content high, dosage is few, the medicine that toxic and side effects is low, by the method such as extracting and developing and purification to Radix Toddaliae Asiaticae, extracts active ingredients, the enrichment of pain easing and hemostasis effect will be had in Radix Toddaliae Asiaticae, toxic component is separated, and by obtained medicinal application in medicament or the medicine of preparing pain easing and hemostasis.
A kind of pain easing and hemostasis of the present invention medicine, be with Radix Toddaliae Asiaticae ( toddalia asiatica(L.) Lam.) root or root bark be raw material, extract through water and alcoholic solution respectively, again through purifying and being separated and obtained a kind of brown solid granule, its main component Coumarins content of chemical substances is not less than 25%, alkaloids substance is lower than 5%, and all the other are mainly the materials such as starch, saccharide, tanning, terpenoid and sterols, and this medicine has good pain easing and hemostasis effect, analgesic effect equals to aspirin, and haemostatic effect is close with YUNNAN BAIYAO.
The coumarin kind compound of above-mentioned indication refers to pimpinellin, different pimpinellin, 8-hydroxyl-6-methoxy coumarin and phellopterin etc.
The alkaloid of above-mentioned indication refers to chelerythrine.
The preparation method of a kind of pain easing and hemostasis of the present invention medicine follows these steps to preparation:
(1) by Radix Toddaliae Asiaticae ( toddalia asiatica(L.) Lam.) root or root bark clean, dry, pulverized 80 mesh sieves;
(2) add water, alcoholic solution respectively: 10%(v/v), 20%(v/v), 30%(v/v), 40%(v/v), 50%(v/v), 60%(v/v), 70%(v/v), 80%(v/v) and 95%(v/v) reflux, extract, 3 times, each 1 hour, solid-liquid ratio first time is 1:12, second and be for three times 1:10;
(3) merged by whole extracting solution, filter, get filtrate decompression and boil off solvent, obtain extraction extractum, in extractum, add the 40-60 DEG C of hot water that 1-2 doubly weighs, stirring suspension, obtains suspension;
(4) successively with the petroleum ether of 0.5-2 times of volume, chloroform, ethyl acetate and n-butanol extraction 1-3 time;
(5) distilling under reduced pressure boils off solvent respectively, obtains petroleum ether extraction extractum, chloroform extraction extractum, extraction into ethyl acetate extractum and n-butanol extraction extractum;
(6) combined chloroform and ethyl acetate layer extractum, is transferred on AB-8 type macroporous adsorptive resins;
(7) respectively with water, 10%(v/v), 20%(v/v), 30%(v/v), 40%(v/v), 50%(v/v), 60%(v/v), 70%(v/v), 80%(v/v) and 95%(v/v) ethanol gradient elution, each gradient elution 5-8 column volume, gets 50%-70% ethanol elution;
(8) 50%-70% ethanol elution is merged, after adopting spray drying technology drying, obtain product.
Extracting mode described in above-mentioned steps (2) also comprises decoction extraction, soxhlet extraction, oozes rumble extraction or supercritical fluid extraction etc. except reflux, extract.
Above-mentioned steps (8) described drying means also comprises room temperature drying under reduced pressure, lyophilization and vacuum microwave drying etc.
The application of a kind of pain easing and hemostasis of the present invention medicine is the medicine for the preparation of pain easing and hemostasis or preparation.
The medicament of above-mentioned indication comprises external pulvis, cataplasma, oral granular formulation, capsule and tablet etc.
The plant amedica extract that pain easing and hemostasis of the present invention medicine can have a function of promoting blood circulation to disperse blood clots with Radix Notoginseng extract or commercially available Radix Notoginseng total arasaponins etc. makes compound pain haemostatic medicament.
Pain easing and hemostasis of the present invention medicine adopts HPLC method to carry out analysis and detects, establish its HPLC finger printing, the content of the Coumarins active component such as pimpinellin, different pimpinellin, phellopterin, Toddalolactone, 8-hydroxyl-6-methoxyl group-coumarin in Simultaneously test medicine, result shows that in medicine, Coumarins composition is not less than 25%, the alkaloid materials such as chelerythrine are lower than 5%, wherein chelerythrine alkali content is lower than 0.5%, significantly reduces toxicant intake.
Pain easing and hemostasis of the present invention medicine adopts docking method to measure the mice bleeding time, shortens to index with the bleeding time before and after mice administration, judges the effect of drug hemostasis.By body weight oral administration gavage administration respectively, every day gavage once, continuous 7 days, adopt capillary glass-tube method to measure clotting time of mice CT after 30 minutes in administration in the 7th day, adopt next day and cut tail method and measure bleeding time BT, shorten to index with clotting time before and after mice administration, judge the effect of hemorrhage, result shows that product of the present invention significantly can shorten bleeding time and the clotting time of mice, shows good styptic activity, shows that it may have facilitation to intrinsic coagulation pathway.Convert average daily people by dosage and be about 5g crude drug with dosage, greatly reduce and take crude drug amount.
The customary adjuvant such as starch, dextrin, carboxymethyl starch sodium, sodium carboxymethyl cellulose, polyvinylpolypyrrolidone, microcrystalline Cellulose, lactose, Pulvis Talci, magnesium stearate are added in the medicine of pain easing and hemostasis of the present invention, conveniently technique, make clinical acceptable dosage form, as external pulvis, cataplasma, oral granular formulation, capsule, pill and tablet etc.
Pain easing and hemostasis of the present invention medicine can also mix in the ratio of 1-100% with the plant amedica extract that Radix Notoginseng extract or commercially available Radix Notoginseng total arasaponins etc. have a function of promoting blood circulation to disperse blood clots, and the crude drug mixed is made external pulvis, cataplasma, oral granular formulation, capsule, pill and tablet etc. by more solito formulation method.
Accompanying drawing explanation
Fig. 1 is the HPLC chromatogram of pain easing and hemostasis of the present invention medicine,
Wherein No. 3 peaks are chlorogenic acid, No. 7 peaks are nitidine, No. 11 peaks are Toddalolactone, and No. 12 peaks are 8-hydroxyl-6-methoxy coumarin, and No. 13 peaks are chelerythrine, No. 14 peaks are .beta.-fagarine, No. 15 peaks are different pimpinellin, and No. 16 peaks are pimpinellin, and No. 20 peaks are phellopterin, Coumarins composition accounts for 35.2% of extract, and the alkaloids substances such as chelerythrine are reduced to less than 2.0%.
Detailed description of the invention
embodiment one (preparation of pain easing and hemostasis medicine):
(1) get root or the root bark 6.0 kilograms of Radix Toddaliae Asiaticae (Toddalia asiatica (L.) Lam.), clean, dry, pulverized 80 mesh sieves;
(2) add water, alcoholic solution respectively: 10%(v/v), 20%(v/v), 30%(v/v), 40%(v/v), 50%(v/v), 60%(v/v), 70%(v/v), 80%(v/v) and 95%(v/v) reflux, extract, 3 times, each 1 hour, solid-liquid ratio first time is 1:12, second and be for three times 1:10;
(3) merged by whole extracting solution, filter, get filtrate decompression and boil off solvent, obtain ethanol extract 1.2 kilograms, in extractum, add 2.5 kilograms of 60 DEG C of hot water, stirring suspension, obtains suspension;
(4) petroleum ether of 2 times of volumes, chloroform, ethyl acetate and n-butanol extraction 3 times are used successively;
(5) distilling under reduced pressure boils off solvent respectively, obtains chloroform extraction extractum 281g, extraction into ethyl acetate extractum 46 and n-butanol extraction extractum 402g;
(6) combined chloroform and ethyl acetate layer extractum, is transferred on AB-8 type macroporous adsorptive resins;
(7) respectively with water, volume ratio 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% and 95% ethanol with the flow rate gradient of 1-3BV/h, each gradient elution 5-8 column volume, gets 50%-70% ethanol elution;
(8) merged by 50%-70% ethanol elution, adopt spray drying technology dry, charging rate 5mL/min, intake air temperature 150-160 DEG C, air outlet temperature 60-70 DEG C, spraying dry obtains product.
embodiment two (this medicine HPLC analyzes):
Pain easing and hemostasis powder described in Example 1 (crossing No. three sieves) carries out HPLC analysis.
(1) chromatographic condition
Chromatographic column Diamonsil C 18(250 mm × 4.6 mm, 5 μm)
Mobile phase A: acetonitrile B:0.75% glacial acetic acid aqueous solution
Type of elution gradient elution
Flow velocity 1.0 mL/min determined wavelength 320 nm
Column temperature 25 DEG C of sample size 20 μ L
The chromatographic run time is 90 minutes, and gradient elution program is in table 1, and under this chromatographic condition, the HPLC chromatogram of pain easing and hemostasis of the present invention medicine is shown in accompanying drawing 1.
(2) sample preparation methods
About extract is about 0.5g, accurately weighed, puts in conical flask, and add 70% EtOH Sonicate and dissolve, filter, filtrate is put in 100 mL measuring bottles, adds 70% ethanol to scale, shakes up, and with 0.45 μm of membrane filtration, to obtain final product.
(3) measurement result
The chromatographic fingerprinting common pattern of Radix Toddaliae Asiaticae extract is set up according to measurement result, 20 total peaks (see accompanying drawing 1) in Radix Toddaliae Asiaticae extract are demarcated, wherein No. 3 peaks are chlorogenic acid, No. 7 peaks are nitidine, No. 11 peaks are Toddalolactone, No. 12 peaks are 8-hydroxyl-6-methoxy coumarin, No. 13 peaks are chelerythrine, No. 14 peaks are .beta.-fagarine, No. 15 peaks are different pimpinellin, No. 16 peaks are pimpinellin, No. 20 peaks are phellopterin, Coumarins composition accounts for 35.2% of extract, the alkaloids substances such as chelerythrine are reduced to less than 2.0%.
embodiment three (after oral medicine mice acetic acid twisting analgesic experiment):
(1) experiment purpose: studied by the analgesic activity of mice acetic acid writhing test to pain easing and hemostasis of the present invention medicine.
(2) experimental subject: Kunming mouse, male and female half and half.
(3) medical material and instrument: pain easing and hemostasis of the present invention medicine is made by oneself by laboratory, and adding distil water makes suspension; Stopwatch.
(4) animal grouping and administration: healthy mice 40 is divided into 4 groups at random, often organize 10, be divided into 3% CMC-Na solution (model group), aspirin group, medicine low dose group of the present invention (amounting to crude drug amount is 0.65g/kg) and medicine high dose group of the present invention (amounting to crude drug amount is 1.3g/kg), every day gavage once, successive administration 5 days, 0.6% acetic acid (0.2ml/ only) is injected in administration in the 5th day 30 minutes pneumoretroperitoneums, observe the writhing number of times that in 20 minutes, each group laboratory animal is caused by acetic acid, result is as table 2:
(5) experimental result: show through one-way ANOVA statistic analysis result: model group, aspirin group and drug administration group mice of the present invention start all to exist between writhing time and writhing number of times significant difference ( p<0.05), illustrate that medicine administration group in pain easing and hemostasis of the present invention all shows analgesic activity in various degree, wherein remarkable with the analgesic activity of extract high dose administration group.
embodiment four (oral drug hemostasis activity experiment):
(1) experiment purpose: adopt docking method to measure the mice bleeding time, shortens to index with the bleeding time before and after mice administration, judges the effect of drug hemostasis.
(2) experimental subject: Kunming mouse, male and female half and half.
(3) animal grouping and administration: get healthy mice 40, be divided into 4 groups at random after raising a week, namely blank group, positive drug group (YUNNAN BAIYAO), medicine low dose group of the present invention and medicine high dose group of the present invention.List of references dosage is also converted according to the dosage of people daily Radix Toddaliae Asiaticae and Yunnan Baiyao Capsule, by body weight respectively by low dose group (amounting to crude drug amount is 0.65g/kg) and high dose group (amounting to crude drug amount is 1.3g/kg) oral administration gavage administration, the dosage of positive drug YUNNAN BAIYAO is 0.2 g/kg, every day gavage once, continuous 7 days, capillary glass-tube method is adopted to measure clotting time of mice after 30 minutes in administration in the 7th day, adopt next day and cut the tail method mensuration bleeding time, the results are shown in Table 3 and table 4.
(4) experimental result: experimental result adopts statistics software SPSS 17.0 to process, and all data show through one-way ANOVA statistic analysis result: blank group and positive controls and all exist between pain easing and hemostasis drug administration group mice bleeding time (BT) value of the present invention and clotting time of mice (CT) significant difference ( p<0.05), illustrate that drug administration group of the present invention all shows the effect shortening mice bleeding time and clotting time in various degree, wherein remarkable with the anastalsis of extract high dose administration group.
embodiment five (preparation of tablet):
Medicine described in Example 1, adopts wet granulation after adding the pharmaceutic adjuvant such as starch, maltose, adds the lubricants such as Pulvis Talci after granulate, according to a conventional method tabletting, obtains pain easing and hemostasis of the present invention sheet.

Claims (9)

1. a pain easing and hemostasis medicine, it is characterized in that with the root of Radix Toddaliae Asiaticae (Toddalia asiatica (L.) Lam.) or root bark for raw material, extract through water and alcoholic solution respectively, again through purifying and being separated and obtained a kind of brown solid granule, its Main Ingredients and Appearance Coumarins content of chemical substances is not less than 25%, alkaloids substance is lower than 5%, all the other are starch, saccharide, tanning, terpenoid and sterols material, this medicine has good pain easing and hemostasis effect, analgesic effect equals to aspirin, and haemostatic effect is close with YUNNAN BAIYAO.
2. a kind of pain easing and hemostasis medicine according to claim 1, is characterized in that the Coumarins chemical substance of indication comprises pimpinellin, different pimpinellin, Toddalolactone, 8-hydroxyl-6-methoxy coumarin and phellopterin.
3. a kind of pain easing and hemostasis medicine according to claim 1, is characterized in that alkaloid refers to chelerythrine toxic biological alkaloid substance.
4. the preparation method of a kind of pain easing and hemostasis medicine according to claim 1, is characterized in that following these steps to preparation:
(1) root of Radix Toddaliae Asiaticae (Toddalia asiatica (L.) Lam.) or root bark are cleaned, dry, pulverized 80 mesh sieves;
(2) add water, alcoholic solution respectively: 10%(v/v), 20%(v/v), 30%(v/v), 40%(v/v), 50%(v/v), 60%(v/v), 70%(v/v), 80%(v/v) and 95%(v/v) reflux, extract, 3 times, each 1 hour, solid-liquid ratio first time is 1:12, second and be for three times 1:10;
(3) merged by whole extracting solution, filter, get filtrate decompression and boil off solvent, obtain ethanol extract, in extractum, add the 40-60 DEG C of hot water that 1-2 doubly weighs, stirring suspension, obtains suspension;
(4) successively with the petroleum ether of 0.5-2 times of volume, chloroform, ethyl acetate and n-butanol extraction 1-3 time;
(5) distilling under reduced pressure boils off solvent respectively, obtains petroleum ether extraction extractum, chloroform extraction extractum, extraction into ethyl acetate extractum and n-butanol extraction extractum;
(6) whole extractum is transferred on AB-8 type macroporous adsorptive resins;
(7) respectively with water, volume ratio 10%, 20%, 30%, 40%, 50%, 60%, 70%, the ethanol gradient elution of 80% and 95%, each gradient elution 5-8 column volume, gets 50%-70% ethanol elution;
(8) 50%-70% ethanol elution is merged, after adopting spray drying technology drying, obtain product.
5. the preparation method of a kind of pain easing and hemostasis medicine according to claim 4, is characterized in that step (2) extracting mode also comprises except reflux, extract, and decocts extractions, soxhlet extraction, oozes rumble extraction or supercritical fluid extraction.
6. the preparation method of a kind of pain easing and hemostasis medicine according to claim 4, is characterized in that step (8) drying means also comprises room temperature drying under reduced pressure, lyophilization and vacuum microwave drying.
7. the application of a kind of pain easing and hemostasis medicine according to claim 1, is characterized in that the medicine for the preparation of pain easing and hemostasis or medicament.
8. the application of a kind of pain easing and hemostasis medicine according to claim 7, is characterized in that the medicament of indication comprises external pulvis, cataplasma, oral granular formulation, capsule or tablet.
9. the application of a kind of pain easing and hemostasis medicine according to claim 7, is characterized in that the plant amedica extract this medicine and Radix Notoginseng extract or commercially available Radix Notoginseng total arasaponins etc. with function of promoting blood circulation to disperse blood clots makes compound pain, haemostatic medicament.
CN201510508482.6A 2015-08-19 2015-08-19 Pain-easing hemostasis medicine and preparation method and application thereof Pending CN105012455A (en)

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Cited By (6)

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CN105640923A (en) * 2016-01-04 2016-06-08 贵阳学院 Novel hemostatic adhesive plaster and preparation method thereof
CN105726802A (en) * 2016-03-01 2016-07-06 南京正亮医药科技有限公司 Preparation method and application of Bazheng tablets
CN107753612A (en) * 2017-10-27 2018-03-06 贵州医科大学 The preparation of rich furocoumarin-containing Radix Toddaliae Asiaticae extract and hemostasis medical usage
CN108042630A (en) * 2018-01-15 2018-05-18 湖北民族学院 A kind of method with HSCCC separation asiatic toddalia root-bark petroleum ether extracts
CN110954643A (en) * 2019-12-20 2020-04-03 成都普思检验检测有限公司 Method for detecting purity of pimpinellide
CN111714686A (en) * 2019-03-19 2020-09-29 广东博与再生医学有限公司 Bone hemostatic material and preparation method thereof

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105640923A (en) * 2016-01-04 2016-06-08 贵阳学院 Novel hemostatic adhesive plaster and preparation method thereof
CN105640923B (en) * 2016-01-04 2018-12-18 贵阳学院 A kind of hemostatic plaster and preparation method thereof
CN105726802A (en) * 2016-03-01 2016-07-06 南京正亮医药科技有限公司 Preparation method and application of Bazheng tablets
CN107753612A (en) * 2017-10-27 2018-03-06 贵州医科大学 The preparation of rich furocoumarin-containing Radix Toddaliae Asiaticae extract and hemostasis medical usage
CN108042630A (en) * 2018-01-15 2018-05-18 湖北民族学院 A kind of method with HSCCC separation asiatic toddalia root-bark petroleum ether extracts
CN111714686A (en) * 2019-03-19 2020-09-29 广东博与再生医学有限公司 Bone hemostatic material and preparation method thereof
CN110954643A (en) * 2019-12-20 2020-04-03 成都普思检验检测有限公司 Method for detecting purity of pimpinellide

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Application publication date: 20151104