CN105596456A - Preparation method and application of spur pills - Google Patents
Preparation method and application of spur pills Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
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- A61K35/56—Materials from animals other than mammals
- A61K35/618—Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
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- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K9/2004—Excipients; Inactive ingredients
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- A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
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- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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Abstract
The invention provides a preparation method of spur pills. The spur pills are prepared from 50 g of kelp, 100 g of rhizome drynariae, 50 g of codonopsis pilosula, 20 g of cassia twig, 100 g of radix clematidis, 50 g of calcined oyster shells, 500 g of folium cortex eucommiae, 100 g of suberect spatholobus stems, 80 g of radix aconiti lateralis praeparata slices, 30 g of radix aconiti preparata, 30 g of prepared kusnezoff monkshood roots, 30 g of processed rhizoma corydalis, 30 g of radix paeoniae alba, 20 g of pseudo-ginseng and 40 g of semen strychni pulveratum by adopting supercritical extraction and microwave extraction, and the drug loading capacity is greatly improved. The invention further provides application of the spur pills for preparing a medicine for constraining rat glioma cell C6 proliferation.
Description
Technical field
The present invention relates to Chinese medicine preparation technical field, be specifically related to a kind of preparation method and application of bony spur tablet.
Background technology
Bony spur tablet is recorded in standards of pharmacopoeia, by kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, the root of Chinese clematis100g, calcined oyster shell 50g, folium cortex eucommiae 500g, reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g,Rhizoma Corydalis (processed) 30g, root of herbaceous peony 30g, pseudo-ginseng 20g, prepared nux vomica 40g make as bulk drug, above ten five tastes, chickenBlood rattan boiling three times, 3 hours for the first time, second and third time each 2 hours, collecting decoction, filtered, and filtrate is for subsequent use;Kelp, the rhizome of davallia, Radix Codonopsis, cassia twig, the root of Chinese clematis, oyster, folium cortex eucommiae boiling secondary, 3 hours for the first time, theSecondary 2 hours, collecting decoction, filters, the merging of filtrate and above-mentioned filtrate, be concentrated into relative density be about 1.35 thickCream; Separately get tag, aconiti preparata,radix, wild aconite root and corydalis tuber four tastes and be ground into meal, soak with acidic ethanol (pH4.5~5.0)Stain is extracted, and filters, and filtrate recycling ethanol, is condensed into thick paste; Root of herbaceous peony Radix Notoginseng powder is broken into fine powder and prepared nux vomica facing-up mixes,Mix with above-mentioned two kinds of thick pastes, dry, pulverize into fine powder, granulate, be pressed into 650, sugar coating, to obtain final product. Loose windHeresy, dispellieg cold and dampness, relaxes the muscles and stimulate the blood circulation, and removes obstruction in channels to relieve pain. For cervical vertebra, chest push away, the hypertrophic teoarthropathy disease such as lumbar vertebrae, calcaneum,Rheumatism, rheumatoid arthritis are had to certain curative effect.
In prior art, not yet there is bony spur tablet adopting the report of overcritical and microwave technology aspect extraction preparation, and adoptThe method that water decocts, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, is inconvenient to take, serious shadowHaving rung this product applies clinically.
Summary of the invention
Goal of the invention: in order to address the above problem, the object of the present invention is to provide a kind of preparation method and bone of bony spur tabletThorn sheet suppresses the application in mouse neuroglial cytoma C6 propagation medicine in preparation.
Technical scheme: the object of the invention is to realize by following scheme:
A preparation method for bony spur tablet, by kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, the root of Chinese clematis100g, calcined oyster shell 50g, folium cortex eucommiae 500g, reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g,Rhizoma Corydalis (processed) 30g, root of herbaceous peony 30g, pseudo-ginseng 20g, prepared nux vomica 40g make as bulk drug, and described method is by followingStep composition: get Radix Codonopsis 50g, cassia twig 20g, tag 80g, join in CO2 supercritical extract device, ethanol is as folderBand agent, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C,CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, for subsequent use; Get in all the otherMedicine, pulverizes, and adds 70% ethanol of 2L, and drop in microwave extracting apparatus and carry out microwave abstracting, extraction power 400-600W,Extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 large pore resin absorption column 50% ethanolWash-out, collects 5 times of amount column volume eluents, and decompression recycling ethanol, concentrates and be dried, and obtains Microwave Extraction thing, for subsequent use;Above-mentioned supercritical extract and Microwave Extraction thing are mixed, add starch, 70% ethanol particle processed, dry, compressing tablet, everySheet 0.5g.
The preparation method of described bony spur tablet, the percent by volume that described CO2 supercritical extract entrainer accounts for total extractant is5%。
The preparation method of described bony spur tablet, the extracting pressure 20MPa of described CO2 supercritical extract, 40 DEG C of temperature, CO2Flow 2ml/g crude drug min, extraction time 160min.
The preparation method of described bony spur tablet, described microwave abstracting power 500W extracts 6 minutes at every turn.
Bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine in preparation, and bony spur tablet is by kelp50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, root of Chinese clematis 100g, calcined oyster shell 50g, folium cortex eucommiae 500g, chickenBlood rattan 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g, Rhizoma Corydalis (processed) 30g, root of herbaceous peony 30g, pseudo-ginseng 20g,Prepared nux vomica 40g makes as bulk drug, and preparation method is made up of the following step: get Radix Codonopsis 50g, cassia twig 20g, attachedSheet 80g, join in CO2 supercritical extract device, ethanol is as entrainer, and entrainer accounts for the volume hundred of total extractantProportion by subtraction is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2 flow 1-3m1/g crude drug min, extractionTime 150-180min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L,Drop in microwave extracting apparatus and carry out microwave abstracting, extraction power 400-600W, extracts 2 times, and each 4-8 minute, closesAnd extract, concentrated, be added on D101 large pore resin absorption column, 50% ethanol elution, collects 5 times of amount column volume wash-outsLiquid, decompression recycling ethanol, concentrates and is dried, and obtains Microwave Extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave are carriedGet thing and mix, add starch, 70% ethanol particle processed, compressing tablet, every 0.5g.
Described bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine, CO2 in preparation method in preparationThe percent by volume that supercritical extract entrainer accounts for total extractant is 5%.
Described bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine, CO2 in preparation method in preparationThe extracting pressure 20MPa of supercritical extract, 40 DEG C of temperature, CO2 flow 2ml/g crude drug min, extraction time 160min.
Described bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine, microwave in preparation method in preparationExtraction power 500W extracts 6 minutes at every turn.
In prior art, every 0.5g of bony spur tablet, each 3,3 times on the one, the bony spur tablet that adopts the present invention to be prepared intoEvery 0.5g,, within 1st, take 3 times, but medicine carrying amount increases greatly by each 3.
Detailed description of the invention
Form by the following examples, is described in further detail foregoing of the present invention again, but should be by thisThe scope that is interpreted as the above-mentioned theme of the present invention only limits to following example, all technology realizing based on foregoing of the present inventionAll belong to scope of the present invention.
Embodiment 1: get kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, root of Chinese clematis 100g, calcined oyster shell50g, folium cortex eucommiae 500g, reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g, Rhizoma Corydalis (processed) 30g,Root of herbaceous peony 30g, pseudo-ginseng 20g, prepared nux vomica 40g, by Radix Codonopsis 50g, cassia twig 20g, tag 80g, to join CO2 superIn critical extractor, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure15MPa, 30 DEG C of temperature, CO2 flow 1m1/g crude drug min, extraction time 150min, obtains supercritical extract,For subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave abstracting, extractionGet power 400W, extract 2 times, each 4 minutes, combining extraction liquid, concentrated, be added to D101 large pore resin absorption columnUpper, 50% ethanol elution, collects 5 times of amount column volume eluents, and decompression recycling ethanol, concentrates and be dried, and obtains microwave and carriesGet thing, for subsequent use; Above-mentioned supercritical extract and Microwave Extraction thing are mixed, add starch, 70% ethanol particle processed, pressesSheet, every 0.5g.
Embodiment 2: get kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, root of Chinese clematis 100g, calcined oyster shell50g, folium cortex eucommiae 500g, reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g, Rhizoma Corydalis (processed) 30g,Root of herbaceous peony 30g, pseudo-ginseng 20g, prepared nux vomica 40g, by Radix Codonopsis 50g, cassia twig 20g, tag 80g, join CO2Super facingIn boundary's extractor, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa,Temperature 50 C, CO2Flow 3m1/g crude drug min, extraction time 180min, obtains supercritical extract, for subsequent use; GetAll the other Chinese medicines, pulverize, and add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave abstracting, extraction power600W, extracts 2 times, and each 8 minutes, combining extraction liquid, concentrated, be added on D101 large pore resin absorption column 50%Ethanol elution, collects 5 times of amount column volume eluents, and decompression recycling ethanol, concentrates and be dried, and obtains Microwave Extraction thing, standbyWith; Above-mentioned supercritical extract and Microwave Extraction thing are mixed, add starch, 70% ethanol particle processed, compressing tablet, every0.5g。
Embodiment 3: get kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, root of Chinese clematis 100g, calcined oyster shell50g, folium cortex eucommiae 500g, reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g, Rhizoma Corydalis (processed) 30g,Root of herbaceous peony 30g, pseudo-ginseng 20g, prepared nux vomica 40g, by Radix Codonopsis 50g, cassia twig 20g, tag 80g, join CO2Super facingIn boundary's extractor, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa,40 DEG C of temperature, CO2Flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, for subsequent use; GetAll the other Chinese medicines, pulverize, and add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave abstracting, extraction power500W, extracts 2 times, and each 6 minutes, combining extraction liquid, concentrated, be added on D101 large pore resin absorption column 50%Ethanol elution, collects 5 times of amount column volume eluents, and decompression recycling ethanol, concentrates and be dried, and obtains Microwave Extraction thing, standbyWith; Above-mentioned supercritical extract and Microwave Extraction thing are mixed, add starch, 70% ethanol particle processed, compressing tablet, every0.5g。
Embodiment 4: bony spur tablet suppresses the experimental study data of mouse neuroglial cytoma C6 propagation
1 experiment material
1.1 experiment cell lines
Mouse neuroglial cytoma C6, Nanjing Medical University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: bony spur tablet of the present invention: press embodiment 3 method preparations.
Liquid liquid storage: take 100mg bony spur tablet, be dissolved in 5ml absolute ethyl alcohol, 0.2 μ m filter filters, 500 μ ldoffPipe packing ,-20 DEG C of storages, 0.2 μ m filter filters the use of absolute ethyl alcohol in order to control group simultaneously.
1.3 experiment reagent
DMEM (Cat.No.12100-061Lot.No.758137 of GIBCO company); Hyclone (Hangzhoupro, sky, ZhejiangThe Lot.No.100419 of bio tech ltd); NaHCO3 (Shanghai hundred million Cat. of chemical reagent Co., Ltd of a specified durationNo.11810-033Lot.No.1088387); Trypsin (AMRESCO company lot number: 2010/04); EDTA(AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt (AMRESCO company lot number:2010242); StreptomycinSulfate (AMRESCO company lot number: 2010382); Absolute ethyl alcohol (NanjingLearn reagent Co., Ltd lot number: 080310182); MTT (Biosharp lot number: 0793); PBS (laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (U.S. MDCompany's model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (Su JingSafe and sound company of group manufactures model: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579);Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi Co., Ltd model:BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air blast is dryDry case (Shanghai accurate experimental facilities company model: DHG9123A); Refrigerator (Siemens Company's model:KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model:KA-1000); 0.2 μ m filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company),96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipette, Tips are some.
2 experimental techniques
1) mouse neuroglial cytoma C6 carries out cellar culture (10cm with DMEM+10%FBS in 37 DEG C, 5%CO2Culture dish), when Growth of Cells is during to logarithmic phase, collecting cell, discards nutrient solution, and PBS fine laundering 3 times, adds 3ml0.25%Trypsase-0.04%EDTA, after 37 DEG C of digestion 2min, adds 5ml complete medium neutralization reaction wherein,After piping and druming cell, proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjusts 3 × 104/ml of concentration of cell suspension.
2) cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, and culture plate is put into cell culture incubator(37 DEG C, 5%CO2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add bony spur tablet solution, continue to cultivate 24h.
4) after 24h, add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT), continue to cultivate 4h.
5) after 4h, buckle method is removed supernatant, pats dry gently with blotting paper, and every hole adds 200 μ l dimethyl sulfoxide (DMSO)s, puts on shaking tableLow-speed oscillation 10min, fully dissolves crystal. Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add nutrient solution) is set simultaneously, control wells (the medicine dissolving medium of cell, same concentrations,Nutrient solution, MTT, dimethyl sulfoxide (DMSO)), set 6 multiple holes for every group.
7) result represents the inhibiting rate of cell with medicine:
Cell increment inhibiting rate (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%. Experiment is heavyMultiple 3 times.
3 statistical dispositions
Adopt correlation analysis and Studentt inspection in MicrosoftExcel2003 software, data are with mean ± S.D.Represent.
4 experimental results
Statistical result showed after mtt assay experiment, with control group comparison, in the time that dosage reaches 5mg/ml, to mouse neurogliaMatter oncocyte C6 propagation suppresses variant (P < 0.05), and dosage this difference in the time of 10mg/ml has conspicuousness (P < 0.01),In the time that reaching 15-20mg/ml, dosage has utmost point significant difference (P < 0.001).
Table 1 bony spur tablet is on mouse neuroglial cytoma C6 cell inhibitory effect impact research (X ± SD)-
Note: with control group comparison, * P < 0.01; * P < 0.001
5 experiment conclusion
Bony spur tablet can suppress mouse neuroglial cytoma C6 propagation, and the cell that reduces mouse neuroglial cytoma C6 is rawLong number order, this effect is dose dependent.
Claims (8)
1. a preparation method for bony spur tablet, by kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, root of Chinese clematis 100g,Calcined oyster shell 50g, folium cortex eucommiae 500g, reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g, Rhizoma Corydalis (processed) 30g,Root of herbaceous peony 30g, pseudo-ginseng 20g, prepared nux vomica 40g make as bulk drug, it is characterized in that described method is made up of the following step:Get Radix Codonopsis 50g, cassia twig 20g, tag 80g, join CO2In supercritical extract device, ethanol is as entrainer, and entrainer accounts forThe percent by volume of total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2Flow 1-3m1/gCrude drug min, extraction time 150-180min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 2L's70% ethanol, drops in microwave extracting apparatus and carries out microwave abstracting, and extraction power 400-600W, extracts 2 times, and each 4-8 dividesClock, combining extraction liquid, concentrated, be added on D101 large pore resin absorption column, 50% ethanol elution, collects 5 times of amount column volumes and washesDe-liquid, decompression recycling ethanol, concentrates and is dried, and obtains Microwave Extraction thing, for subsequent use; By above-mentioned supercritical extract and Microwave ExtractionThing mixes, and adds starch, and 70% ethanol particle processed is dry, compressing tablet, every 0.5g.
2. the preparation method of bony spur tablet according to claim 1, is characterized in that described CO2Supercritical extract entrainer accounts forThe percent by volume of total extractant is 5%.
3. the preparation method of bony spur tablet according to claim 1, is characterized in that described CO2The extraction of supercritical extract is pressedPower 20MPa, 40 DEG C of temperature, CO2Flow 2ml/g crude drug min, extraction time 160min.
4. the preparation method of bony spur tablet according to claim 1, is characterized in that described microwave abstracting power 500W, eachExtract 6 minutes.
5. bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine in preparation, it is characterized in that bony spur tabletBy kelp 50g, rhizome of davallia 100g, Radix Codonopsis 50g, cassia twig 20g, root of Chinese clematis 100g, calcined oyster shell 50g, folium cortex eucommiae 500g,Reticulate millettia 100g, tag 80g, aconiti preparata,radix 30g, wild aconite root 30g, Rhizoma Corydalis (processed) 30g, root of herbaceous peony 30g, pseudo-ginseng 20g, horseMoney powder 40g makes as bulk drug, and preparation method is made up of the following step: get Radix Codonopsis 50g, cassia twig 20g, tag 80g,Join CO2In supercritical extract device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%,Extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2Flow 1-3m1/g crude drug min, extraction time 150-180min,Obtain supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatusRow microwave abstracting, extraction power 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added to D101On large pore resin absorption column, 50% ethanol elution, collects 5 times of amount column volume eluents, and decompression recycling ethanol, concentrates and be dried,Obtain Microwave Extraction thing, for subsequent use; Above-mentioned supercritical extract and Microwave Extraction thing are mixed, add starch, 70% ethanol particle processed,Dry, compressing tablet, every 0.5g.
6. bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine in preparation according to claim 5, itsBe characterised in that CO in preparation method2The percent by volume that supercritical extract entrainer accounts for total extractant is 5%.
7. bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine in preparation according to claim 5, itsBe characterised in that CO in preparation method2The extracting pressure 20MPa of supercritical extract, 40 DEG C of temperature, CO2Flow 2ml/g crude drugMin, extraction time 160min.
8. bony spur tablet suppresses the application in mouse neuroglial cytoma C6 propagation medicine in preparation according to claim 5, itsBe characterised in that, in preparation method, microwave abstracting power 500W extracts 6 minutes at every turn.
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CN101897770A (en) * | 2010-08-10 | 2010-12-01 | 浙江维康药业有限公司 | Bone spur capsule and preparation process thereof |
CN102988501A (en) * | 2012-10-08 | 2013-03-27 | 卞毓平 | Preparation method and application of Libiling tablet |
CN102988500A (en) * | 2012-10-08 | 2013-03-27 | 卞毓平 | Method for preparing compound Dantong tablets |
CN103467480A (en) * | 2013-10-10 | 2013-12-25 | 南京正亮医药科技有限公司 | Method for extracting artemisinin and application of artemisinin |
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2016
- 2016-03-01 CN CN201610116266.1A patent/CN105596456A/en not_active Withdrawn
Patent Citations (4)
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CN101897770A (en) * | 2010-08-10 | 2010-12-01 | 浙江维康药业有限公司 | Bone spur capsule and preparation process thereof |
CN102988501A (en) * | 2012-10-08 | 2013-03-27 | 卞毓平 | Preparation method and application of Libiling tablet |
CN102988500A (en) * | 2012-10-08 | 2013-03-27 | 卞毓平 | Method for preparing compound Dantong tablets |
CN103467480A (en) * | 2013-10-10 | 2013-12-25 | 南京正亮医药科技有限公司 | Method for extracting artemisinin and application of artemisinin |
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Title |
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刘锡钧: "《福建省基本医疗保险药品实用指南》", 31 March 2002 * |
陈功锡: "《湘西药用植物资源开发与可持续利用》", 31 May 2015 * |
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