CN103432181A - Common pternopetalum root extract and new use thereof in pharmacy - Google Patents
Common pternopetalum root extract and new use thereof in pharmacy Download PDFInfo
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- CN103432181A CN103432181A CN2013103774226A CN201310377422A CN103432181A CN 103432181 A CN103432181 A CN 103432181A CN 2013103774226 A CN2013103774226 A CN 2013103774226A CN 201310377422 A CN201310377422 A CN 201310377422A CN 103432181 A CN103432181 A CN 103432181A
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Abstract
The invention relates to a new use of a common pternopetalum root extract, and particularly relates to the use of the common pternopetalum root extract in the field of pharmacy. The use concretely comprises the functions of treating and preventing dysmenorrheal, treating and preventing hypertension, treating and preventing atherosclerosis, treating and preventing cerebral thrombosis and the like caused by cerebral apoplexy. The common pternopetalum root extract is safe and non-toxic, and strong in pharmacologic action, and heralds a good medicinal prospect; meanwhile, the common pternopetalum root extract is simple in preparation process, and can be prepared into capsules, tablets, cream and oiling agents to be taken orally or used externally.
Description
Technical field
The present invention relates to a kind of new purposes of extractive of Zi-Jin-Sha, relate in particular to the purposes in pharmaceutical field.
Background technology
CN101002809A discloses the extractive of Zi-Jin-Sha bisabolangelone, and its structural formula is:
Extract in the root that this compound or mixture can turn celery Angelica polymorpha Maxim. by the Umbelliferae archangel, be distributed in Hubei, Hunan, Chongqing etc. and economize ground.Effect with warming spleen and stomach for dispelling cold, regulating QI to relieve pain.Known can be used as treating stomachache, stomachache, chest and hypochondrium, rheumatic arthritis, traumatic injury, venom etc.
Summary of the invention
The object of the present invention is to provide extractive of Zi-Jin-Sha and new purposes, i.e. new application in pharmacy.
The present invention realizes above-mentioned purpose like this: a kind of extractive of Zi-Jin-Sha, it is basic raw material that the samphire of take is turned celery (Angelica polymorpha Maxim.), ethanol is extracting solution, and concentrating under reduced pressure obtains dark-brown extractum extract, then extracts and make successively through organic solvent.Described concentration of alcohol is 40-100%, and in ethanol extraction, the percentage by weight of bisabolangelone is greater than 2%.Described organic solution is respectively the petroleum ether of 95-98%, the chloroform of 95-98%, the ethyl acetate of 95-98%, the n-butyl alcohol of 95-98%, by organic solution ethanol extraction extract successively, obtains respectively petroleum ether part, chloroform extract, ethyl acetate extract, n-butanol portion.Get ethyl acetate extract, often pressure column chromatography, take volume ratio as 5-8:1 with petroleum ether and ethyl acetate, eluting repeatedly, and recrystallization obtains the compound bisabolangelone.
The invention further relates to extractive of Zi-Jin-Sha as the new purposes in the medicine of preparation treatment and prevention dysmenorrhea.
Relate to extractive of Zi-Jin-Sha as the new purposes in the medicine of preparation treatment and prophylaxis of hypertension.
Relate to extractive of Zi-Jin-Sha as the new purposes in the medicine that preparation is treated and prevention of arterial is atherosis.
Relate to the new purposes in the medicine of the cerebral thrombosis that extractive of Zi-Jin-Sha causes as preparation treatment and preventing brain stroke.
The invention has the advantages that:
1) the present invention, to extractive of Zi-Jin-Sha and main constituent bisabolangelone, has excavated new medical usage, has opened up new application.
2) extractive of Zi-Jin-Sha safety non-toxic of the present invention, pharmacological action is strong, is indicating well prospect in medicine.
3) extractive of Zi-Jin-Sha raw material sources of the present invention abundant, inexpensive, have no toxic and side effects, preparation technology is simple, can make capsule, tablet, ointment, oil preparation for for oral administration or external.
4) extractive of Zi-Jin-Sha of the present invention has the effect of obvious treatment dysmenorrhea, and it has obviously reduced the contraction frequency that oxytocin causes, mean tension, average amplitude; Reduced CaCl
2the contraction frequency caused, mean tension, average amplitude; The mean tension that KCl causes, average amplitude and tension force maximum have obviously been reduced simultaneously; Contraction frequency, mean tension and average amplitude that rat causes in the oxytocin of body uterus have also obviously been reduced.
5) extractive of Zi-Jin-Sha of the present invention has the diseases such as cerebral ischemia that hypertension, atherosclerosis, apoplexy that obvious treatment is relevant to calcium antagonist cause.
The accompanying drawing explanation
The HPLC of Fig. 1 Radix Pternopetali vulgaris ethanol extraction analyzes collection of illustrative plates: wherein, and chromatographic column XB-C
18(4.6mm * 250mm, 5 μ m); Detect wavelength 248nm; 25 ℃ of column temperatures; Mobile phase: (A) acetonitrile, (B) water, gradient elution, 0～23min (15%～55%A, 45%～85%B), 23～35min (55%～70%A, 30%～45%B), 35～45min (70%～95%A, 5%～30%B), 45～50min (95%A, 5%B); Sample size is 10 μ L, and flow velocity is 0.8mL/min.
The HPLC of Fig. 2 bisabolangelone analyzes collection of illustrative plates.
The HPLC of Fig. 3 petroleum ether part analyzes collection of illustrative plates.Wherein, main special characteristic component is: aqua oxidation peucedanin (retention time: 17.5min), bisabolangelone (retention time: 27min) and isoimperatorin (retention time: 36.5min).
The HPLC of Fig. 4 chloroform extract analyzes collection of illustrative plates.
The HPLC of Fig. 5 ethyl acetate extract analyzes collection of illustrative plates.
The HPLC of Fig. 6 n-butanol portion analyzes collection of illustrative plates.
The specific embodiment
In order to understand better essence of the present invention, below will its new purposes in pharmaceutical field be described by pharmacological testing and the result of extractive of Zi-Jin-Sha.
Take 3 kilograms, Radix Pternopetali vulgaris powder (40 order), with 95% ethanol difference atmospheric pressure reflux, extract three times, merge all extracting solution, reclaim solvent, concentrating under reduced pressure obtains dark-brown extractum, with appropriate distilled water, extractum is suspended, extracted with petroleum ether, chloroform, ethyl acetate, water saturated n-butyl alcohol successively.Add 9500mL extract and 500mL water to mix, standing 12h, after two phase stratification, utilizes the siphonage principle that the extract on upper strata is separated out, and concentrates and reclaim extract on Rotary Evaporators at every turn, and the solvent after recovery continues on for extraction.The extract that merges each extractant volatilizes solvent in fume hood, finally: petroleum ether part, chloroform extract, ethyl acetate extract, n-butanol portion, remaining aqueous solution part get Shui position after the lyophilizing of frozen drying instrument, save backup.
Get ethyl acetate extract and carry out in right amount the normal pressure silica gel column chromatography, take petroleum ether-ethyl acetate (volume ratio is 6:1) eluting repeatedly, recrystallization obtains the compound bisabolangelone.Controlling the bisabolangelone dosage is 1-10mg/kg, makes capsule, tablet, ointment, oil preparation for for oral administration or external.
1. the research of extractive of Zi-Jin-Sha, the effect of bisabolangelone treatment dysmenorrhea
1.1 the impact that extractive of Zi-Jin-Sha, bisabolangelone shrink isolated uterine
Continuous three days subcutaneous injection estradiol benzoates (1mg/kg) before the pregnant mouse experiment, do not make animal in rutting period, to improve the sensitivity of uterine smooth muscle to medicine.Mice is put to death in the cervical vertebra dislocation, and clip one Aconitum carmichaeli Debx. palace (about 2cm is long), be placed in the glass dish that fills Rockwell liquid immediately rapidly, residual connective tissue and the fat of careful removing, two ends are pricked with toe-in respectively, and an end is fixed in the isolated smooth muscle bath, and the other end is connected with tonotransducer.Add 16ml Rockwell liquid in groove, pass into continuously 95%O
2add 5%CO
2mist, constant temperature (37 ± 0.5) ℃, load 1g.After the easypro contracting in uterus is stable, record one section normal contraction activity curve.
(1) extractive of Zi-Jin-Sha, bisabolangelone cause on oxytocin the impact that Mouse Uterus is shunk
After the specimen balance, to adding oxytocin to make final concentration in bath, be 0.01U/ml, the shrinkage curve of one section isolated uterine of record in contrast, after changing clothes nutritional solution in bath 3 times, the oxytocin that adds same amount, after curve shrinkage is highly stable, adding different extractive of Zi-Jin-Sha to make final concentration is 1.6 μ g/ml, 3.2 μ g/ml, the final concentration of bisabolangelone is 0.8 μ g/m, 1.2 μ g/ml, 1.6 μ g/ml, record each cross-talk palace shrinkage curve, statistics contraction frequency, mean tension, average amplitude, the results are shown in Table 1.
Table 1 extractive of Zi-Jin-Sha, bisabolangelone cause on oxytocin the impact that Mouse Uterus is shunk
Compare * p<0.05 with matched group; * p<0.01 area=average amplitude * time
Above-mentioned result of the test shows, the Radix Pternopetali vulgaris petroleum ether part of variable concentrations, chloroform extract, ethyl acetate extract, and the bisabolangelone of variable concentrations has all obviously reduced the contraction frequency that oxytocin causes, mean tension and average amplitude.And, along with the increase of Drug level, the reduction degree strengthens.
(2) extractive of Zi-Jin-Sha, bisabolangelone are to CaCl
2the uterotonic impact caused
After the specimen balance is good, first uses without the calcium High potassium solution and rinse 3 times, then in without the calcium High potassium solution depolarization 30min, add CaCl
2making final concentration is 125 μ g/ml, records CaCl
2the shrinkage curve in the uterus caused in contrast.After without the calcium High potassium solution, changing clothes 3 times, first adding different extractive of Zi-Jin-Sha to make final concentration is 1.6 μ g/ml, 3.2 μ g/ml, and the final concentration of bisabolangelone is 0.8 μ g/ml, 1.2 μ g/ml, 1.6 μ g/ml, then adds the CaCl of same amount
2, record shrinkage curve the relatively variation of front and back, the results are shown in Table 2.
Table 2 extractive of Zi-Jin-Sha, bisabolangelone cause on CaCl2 the impact that Mouse Uterus is shunk
Compare * p<0.05 with matched group; * p<0.01 area=average amplitude * time
Above-mentioned result of the test shows, the Radix Pternopetali vulgaris petroleum ether part of variable concentrations, chloroform extract, ethyl acetate extract, and the bisabolangelone of variable concentrations has all obviously reduced CaCl
2the contraction frequency caused, mean tension and average amplitude.And, along with the increase of Drug level, the reduction degree strengthens.This effect and inhibition CaCl
2overflow is relevant.
(3) the uterotonic impact that extractive of Zi-Jin-Sha, bisabolangelone cause High K+ depolarization
After the specimen balance is good, adding KCl to make final concentration is 7.5 * 10
3μ g/ml, record shrinkage curve in contrast, after changing clothes nutritional solution 3 times, first adding different extractive of Zi-Jin-Sha to make final concentration is 1.6 μ g/ml, 3.2 μ g/ml, the final concentration of bisabolangelone is 0.8 μ g/ml, 1.2 μ g/ml, 1.6 μ g/ml, add the KCl of same amount to record shrinkage curve, relatively the contraction change of front and back, the results are shown in Table 3 again.
Table 3 extractive of Zi-Jin-Sha, bisabolangelone cause on High K+ depolarization the impact that Mouse Uterus is shunk
Compare * p<0.05 with matched group; * p<0.01 area=average amplitude * time
Above-mentioned result of the test shows, the Radix Pternopetali vulgaris petroleum ether part of variable concentrations, chloroform extract, ethyl acetate extract, and the bisabolangelone of variable concentrations has all obviously reduced the mean tension that KCl causes, average amplitude and tension force maximum.And, along with the increase of Drug level, the reduction degree strengthens.In the calcium that this effect causes with the inhibition High K+ depolarization, stream is relevant.
1.2 extractive of Zi-Jin-Sha, the bisabolangelone impact on oxytocin induced mice pain
Get Kunming kind female mice, random packet after the title quality status stamp, 10 every group.Manufacture the primary dysmenorrhea model with estradiol benzoate and oxytocin.Press 10mg/kg lumbar injection estradiol benzoate injection, continuous 5d, to increase the sensitivity of uterine smooth muscle to medicine.The while gastric infusion, 1h after the last administration, lumbar injection oxytocin 30U/kg, the index that the writhing response of take shrinks strongly as uterus, writhing incidence rate and the number of times of the interior mice of the rear 30min of record injection, the results are shown in Table 4.
The impact on oxytocin induced mice pain writhing of table 4 extractive of Zi-Jin-Sha, bisabolangelone
Compare * P<0.05 with model group; * p<0.01 area=average amplitude * time
Above-mentioned result of the test shows, the Radix Pternopetali vulgaris petroleum ether part of various dose, and chloroform extract, ethyl acetate extract, bisabolangelone all obviously reduce the mouse writhing reaction times.Show that it has the effect for the treatment of dysmenorrhea.
1.3 extractive of Zi-Jin-Sha, bisabolangelone are on rat strong impact of shrinking in the body uterus due to oxytocin
Laboratory animal SD rat, female, body weight 230～250g is provided by SanXia University's zoopery center:
(1) animal grouping: get 36 of healthy adult female sd inbred rats, 3d is fed and observed to adaptability to confirm health.Be divided at random 12 groups, 3 every group, i.e. matched group, bisabolangelone group, Radix Pternopetali vulgaris petroleum ether part group, ethyl acetate extract group, chloroform extract group, n-butanol portion group.
(2) animals administer and nursing: each organizes and regularly gives gavage 2 times every day, bisabolangelone by 8,10, the 12mg/kg administration, Radix Pternopetali vulgaris petroleum ether part group, ethyl acetate extract group, chloroform extract group, the n-butanol portion group by 10, the 14mg/kg administration, matched group gives the normal saline of equal volume, continuous use 7d, 8d morning, after the last medicine, row is tested in the body uterus half an hour, and 3d starts subcutaneous injection diethylstilbestrol injection 2mg/ (kgd) before experiment, be used in conjunction 2d, to improve the sensitivity of uterine smooth muscle.
(3) rat is made at the body model of uterus: every group of gavage 8d, the rear use 20% urethane intraperitoneal injection of anesthesia (1.3g/kg) of weighing, the rat dorsal position is fixed on the Mus plate, opens abdomen and find out a side cornua uteri, ligation dialysis ovary, the blunt separation surrounding tissue, the uterus far-end is drawn in to uterus heat-preservation cylinder bottom outlet, clamp gently with the frog heart clip that is connected with cotton thread, connect tension pick-up, fixedly the uterus heat-preservation cylinder, in the position of rat abdomen, is transferred to the heat-preservation cylinder middle part by this position, Aconitum carmichaeli Debx. palace.And added the De Shi liquid of 37 ℃～38 ℃ by place, hole on heat-preservation cylinder, and whole uterus is immersed in De Shi liquid, irradiate insulation with operating lamp.The bio signal analytical system starts to record the uterine activity curve, recording curve 20～30min after uterine contraction is steady.Wait shrink curve steadily after intramuscular injection oxytocin (20U/kg) record frequency, mean tension and the area that rat uterus shrinks, the results are shown in Table 5.
Table 5 extractive of Zi-Jin-Sha, bisabolangelone cause rat strong impact of shrinking in the body uterus to oxytocin
Compare * p<0.05 with matched group; * p<0.01 area=average amplitude * time
Above-mentioned result of the test shows, Radix Pternopetali vulgaris petroleum ether part, ethyl acetate extract, chloroform extract, and the bisabolangelone of variable concentrations has all obviously reduced contraction frequency, mean tension and average amplitude that rat causes in the oxytocin of body uterus.
1.4 the observation of Radix Pternopetali vulgaris ethanol extract treatment dysmenorrhea effect
Clinical data: this is organized 60 routine patients and is the out-patient of Yichang City institute of traditional Chinese medicine, and wherein middle school student account for the overwhelming majority.Unmarried 49 examples, married 11 examples; The oldest person 38 years old, age reckling 14 years old; The course of disease is long 11 years, and the course of disease is short 7 months; This is organized 60 routine patients and is without organic disease.
The new fresh ginger of Therapeutic Method: the 10g filtered through gauze of squeezing the juice, add the 40mg ethanol extract and mix thoroughly, on the coating gauze, is affixed on SHENQUE acupoint, fixing.Once a day.Treatment time is through about the first seven day, treats continuously three menstrual period, if invalid, stops treatment.
The treatment standard: it is recovery from illness that pain disappears without the recidivist; Pain obviously alleviates or attack times is reduced to effectively; Treat three through after date, pain is invalid before treating.
Therapeutic outcome: this organizes 60 examples, and 33 examples of fully recovering, account for 55%; Effective 25 examples, account for 41.7%; Invalid 2 examples, account for 3.3%; Total effective rate 96.7%.
Experimental result shows that Radix Pternopetali vulgaris has the effect of obvious treatment dysmenorrhea, ethanol extract extraction position PetroChina Company Limited. ether position, and chloroform extract, ethyl acetate extract is effective, and bisabolangelone is main effective ingredient.Radix Pternopetali vulgaris suppresses uterine contraction, has the effect of prevention and treatment dysmenorrhea.
Embodiment 2
Extractive of Zi-Jin-Sha, the bisabolangelone Imaging in Patients with Cerebral Ischemia Disease that hypertension, atherosclerosis, the apoplexy relevant to calcium antagonist with prevention causes in treatment.
2.1 calcium antagonism
Uterine Smooth Cell is cultivated:
Mouse anesthesia is put to death, take out rapidly uterus and be put in pre-cooling Hanke, in s liquid (0-4 ℃), rinse 2-3 time, with shears, surrounding tissue is removed totally, perimetrium is wiped off gently with blade, tiling is vertically cutd open in uterus, lining endothelium is fixed an end with tweezers upward, and the syringe needle (bending into 30 degree angles) started with disposable syringe from bearing is partly wiped endometrium and substrate off gently as separator, inner membrance and substrate is removed to clean uterine smooth muscle bar and shred to 1mm
3collagenase and 37 ℃ of water-bath concussion digestion 1h of 0.25% pancreatin mixture slaking liquid with 3ml0.25%, there is serum DMEM-F12 culture fluid to stop digestion, for several times, 200 order cell sieves filter draws suspension in piping and druming, then by 400 order cell sieves, collect filtrate 1000rpm10min centrifugal, basic culture solution is washed 2 times, removes cell debris, is inoculated in the 35mm culture dish and cultivates.Platform is expected and blue repelled test and look into cell survival rate and be greater than 90%, uses the 20%FCSDMEM culture fluid, puts into 37 ℃, 95% humidity, 5%CO
2incubator in cultivate, changed a not good liquor every 3 days.
2.1.1 extractive of Zi-Jin-Sha, the bisabolangelone impact on KCl-induced Uterine Smooth Cell calcium ion concentration
After uterine cell at room temperature being used to the Fluo-3AM dyeing 60min of 100mmol/L esterified form, by the Hank immersion, wash away extracellular residual fluorescence stain, select active good cell and tested under phase contrast microscope.In 40 orifice plates, active good cell is divided into 12 groups at random: matched group, the basic, normal, high concentration group of bisabolangelone, extractive of Zi-Jin-Sha is low, the high concentration group, with the LSM-510 laser confocal microscope, measures the reflection Free Ca
2+intracellular Ca2+ fluorescence intensity (the excitation wavelength 488nm of concentration, emission wavelength 505nm), after measuring the basic value of calcium fluorescence intensity, give respectively Hank ' S liquid, final concentration is 0.8 μ g/ml, 1.2 μ g/ml, 1.6 μ g/ml bisabolangelones, 1.6 μ g/ml, 3.2 μ g/ml extractive of Zi-Jin-Sha are measured the calcium fluorescence intensity after processing 20min again, then add potassium chloride to scan a width every 10s and record calcium fluorescence intensity peak value simultaneously.Each orifice plate is random selected 10 cells under laser confocal microscope, and calcium fluorescence intensity in analysis of cells is averaged after recording peak value, the results are shown in Table 6.
The impact on KCl-induced Uterine Smooth Cell calcium ion fluorescence intensity peak change of table 6 extractive of Zi-Jin-Sha and bisabolangelone
Compare * p<0.05 with matched group; * p<0.01
2.1.2 extractive of Zi-Jin-Sha, bisabolangelone bring out the impact of Uterine Smooth Cell calcium ion concentration on caffeine
The same 2.1.1 of experimental technique, replace potassium chloride with caffeine (final concentration 20mmol/L), the results are shown in Table 7.
Table 7 bisabolangelone and Radix Pternopetali vulgaris different parts extract bring out the impact of Uterine Smooth Cell calcium ion fluorescence intensity peak change on caffeine
Compare * p<0.05 with matched group; * p<0.01
Above-mentioned result of the test shows, in the outer calcium that the bisabolangelone of variable concentrations, Radix Pternopetali vulgaris petroleum ether part, chloroform extract, ethyl acetate extract are not only induced potassium chloride, stream has blocking effect, and the interior calcium that caffeine is induced discharges certain blocking effect is also arranged, proof bisabolangelone and petroleum ether part, chloroform extract, ethyl acetate extract extract have calcium antagonism, the cerebral ischemia that effectively treatment and prophylaxis of hypertension, atherosclerosis, apoplexy cause.
Claims (9)
1. an extractive of Zi-Jin-Sha is characterized in that: with samphire turn celery (
angelica polymorphamaxim.) be basic raw material, ethanol is extracting solution, and concentrating under reduced pressure obtains dark-brown extractum extract, then extracts and make successively through organic solvent.
2. extractive of Zi-Jin-Sha according to claim 1, it is characterized in that: concentration of alcohol is 40-100%, in ethanol extraction, the percentage by weight of bisabolangelone is greater than 2%.
3. extractive of Zi-Jin-Sha according to claim 1, it is characterized in that: described organic solution is respectively the petroleum ether of 95-98%, the chloroform of 95-98%, the ethyl acetate of 95-98%, the n-butyl alcohol of 95-98%, by organic solution ethanol extraction extract successively, obtains respectively petroleum ether part, chloroform extract, ethyl acetate extract, n-butanol portion.
4. extractive of Zi-Jin-Sha according to claim 1 is characterized in that: get ethyl acetate extract, often pressure column chromatography, take volume ratio as 5-8:1 with petroleum ether and ethyl acetate, eluting repeatedly, and recrystallization obtains compound bisabolane Ji copper.
5. the new purposes of the described extractive of Zi-Jin-Sha of claim 1-4 any one in the medicine of preparation treatment and prevention dysmenorrhea.
6. the purposes of the described extractive of Zi-Jin-Sha of claim 1-4 any one in the medicine of preparation treatment and prophylaxis of hypertension.
7. the purposes of the described extractive of Zi-Jin-Sha of claim 1-4 any one in preparation treatment and the atherosis medicine of prevention of arterial.
8. the purposes in the medicine of the cerebral thrombosis that the described extractive of Zi-Jin-Sha of claim 1-4 any one causes at preparation treatment and preventing brain stroke.
9. the purposes of the described extractive of Zi-Jin-Sha of claim 5-8 any one in preparing medicine, it is characterized in that: the bisabolangelone dosage is 1-10mg/kg, can be made into capsule, tablet, ointment, oil preparation for for oral administration or external.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004002503A1 (en) * | 2002-06-27 | 2004-01-08 | Chinese Medicines Scientific Consultants Pty Ltd | Compositions and methods for treating gynaecological disorders |
| CN103156893A (en) * | 2011-12-09 | 2013-06-19 | 中国人民解放军军事医学科学院毒物药物研究所 | Novel application for angelica polymorpha maxim and extract thereof |
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2013
- 2013-08-27 CN CN201310377422.6A patent/CN103432181B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004002503A1 (en) * | 2002-06-27 | 2004-01-08 | Chinese Medicines Scientific Consultants Pty Ltd | Compositions and methods for treating gynaecological disorders |
| CN103156893A (en) * | 2011-12-09 | 2013-06-19 | 中国人民解放军军事医学科学院毒物药物研究所 | Novel application for angelica polymorpha maxim and extract thereof |
Non-Patent Citations (2)
| Title |
|---|
| 万威: "土家族药紫金砂化学成分及质量标准研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》, 15 November 2009 (2009-11-15), pages 056 - 997 * |
| 惠永正: "《中药天然产物大全 11 下 中药》", 31 January 2011, article "拐芹" * |
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