CN103751701A - Danshilitong tablet preparation method and application of Danshilitong tablets in preparation of lymphoma cell YAC-1 proliferation inhibition drugs - Google Patents
Danshilitong tablet preparation method and application of Danshilitong tablets in preparation of lymphoma cell YAC-1 proliferation inhibition drugs Download PDFInfo
- Publication number
- CN103751701A CN103751701A CN201310672109.5A CN201310672109A CN103751701A CN 103751701 A CN103751701 A CN 103751701A CN 201310672109 A CN201310672109 A CN 201310672109A CN 103751701 A CN103751701 A CN 103751701A
- Authority
- CN
- China
- Prior art keywords
- preparation
- extraction
- danshilitong
- cholelithiasis treating
- danshitongli tablet
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 38
- 239000003814 drug Substances 0.000 title claims abstract description 25
- 229940079593 drug Drugs 0.000 title claims abstract description 16
- 101000981253 Mus musculus GPI-linked NAD(P)(+)-arginine ADP-ribosyltransferase 1 Proteins 0.000 title claims abstract description 14
- 206010025323 Lymphomas Diseases 0.000 title claims abstract description 9
- 230000005764 inhibitory process Effects 0.000 title abstract description 3
- 230000035755 proliferation Effects 0.000 title abstract 2
- 241000717739 Boswellia sacra Species 0.000 claims abstract description 17
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 17
- 238000000874 microwave-assisted extraction Methods 0.000 claims abstract description 11
- 239000011347 resin Substances 0.000 claims abstract description 7
- 229920005989 resin Polymers 0.000 claims abstract description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 57
- 201000001883 cholelithiasis Diseases 0.000 claims description 35
- 238000000605 extraction Methods 0.000 claims description 27
- 239000004863 Frankincense Substances 0.000 claims description 16
- 241001057584 Myrrha Species 0.000 claims description 16
- RSIJVJUOQBWMIM-UHFFFAOYSA-L sodium sulfate decahydrate Chemical compound O.O.O.O.O.O.O.O.O.O.[Na+].[Na+].[O-]S([O-])(=O)=O RSIJVJUOQBWMIM-UHFFFAOYSA-L 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 9
- 230000004663 cell proliferation Effects 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 5
- 230000000274 adsorptive effect Effects 0.000 claims description 5
- 239000012141 concentrate Substances 0.000 claims description 5
- 230000006837 decompression Effects 0.000 claims description 5
- 239000003480 eluent Substances 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 5
- 239000012567 medical material Substances 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 238000004064 recycling Methods 0.000 claims description 5
- 239000008107 starch Substances 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 235000019640 taste Nutrition 0.000 claims description 5
- 239000001100 (2S)-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chroman-4-one Substances 0.000 abstract description 12
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 abstract description 12
- QUQPHWDTPGMPEX-UTWYECKDSA-N aurantiamarin Natural products COc1ccc(cc1O)[C@H]1CC(=O)c2c(O)cc(O[C@@H]3O[C@H](CO[C@@H]4O[C@@H](C)[C@H](O)[C@@H](O)[C@H]4O)[C@@H](O)[C@H](O)[C@H]3O)cc2O1 QUQPHWDTPGMPEX-UTWYECKDSA-N 0.000 abstract description 12
- APSNPMVGBGZYAJ-GLOOOPAXSA-N clematine Natural products COc1cc(ccc1O)[C@@H]2CC(=O)c3c(O)cc(O[C@@H]4O[C@H](CO[C@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)cc3O2 APSNPMVGBGZYAJ-GLOOOPAXSA-N 0.000 abstract description 12
- VUYDGVRIQRPHFX-UHFFFAOYSA-N hesperidin Natural products COc1cc(ccc1O)C2CC(=O)c3c(O)cc(OC4OC(COC5OC(O)C(O)C(O)C5O)C(O)C(O)C4O)cc3O2 VUYDGVRIQRPHFX-UHFFFAOYSA-N 0.000 abstract description 12
- QUQPHWDTPGMPEX-QJBIFVCTSA-N hesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]4[C@@H]([C@H](O)[C@@H](O)[C@H](C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-QJBIFVCTSA-N 0.000 abstract description 12
- 229940025878 hesperidin Drugs 0.000 abstract description 12
- ARGKVCXINMKCAZ-UHFFFAOYSA-N neohesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)OC3C(C(O)C(O)C(C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UHFFFAOYSA-N 0.000 abstract description 12
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 2
- 235000008658 Artemisia capillaris Nutrition 0.000 abstract 1
- 241000092668 Artemisia capillaris Species 0.000 abstract 1
- 235000003717 Boswellia sacra Nutrition 0.000 abstract 1
- 241000675108 Citrus tangerina Species 0.000 abstract 1
- 240000007311 Commiphora myrrha Species 0.000 abstract 1
- 235000006965 Commiphora myrrha Nutrition 0.000 abstract 1
- 244000164480 Curcuma aromatica Species 0.000 abstract 1
- 235000003398 Curcuma aromatica Nutrition 0.000 abstract 1
- 241000506964 Dichondra repens Species 0.000 abstract 1
- 244000111489 Gardenia augusta Species 0.000 abstract 1
- 235000018958 Gardenia augusta Nutrition 0.000 abstract 1
- 244000303040 Glycyrrhiza glabra Species 0.000 abstract 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 abstract 1
- 235000017443 Hedysarum boreale Nutrition 0.000 abstract 1
- 235000007858 Hedysarum occidentale Nutrition 0.000 abstract 1
- 235000007265 Myrrhis odorata Nutrition 0.000 abstract 1
- 241000219061 Rheum Species 0.000 abstract 1
- 240000004534 Scutellaria baicalensis Species 0.000 abstract 1
- 235000017089 Scutellaria baicalensis Nutrition 0.000 abstract 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 abstract 1
- 241000329195 Sparganium erectum Species 0.000 abstract 1
- 238000010521 absorption reaction Methods 0.000 abstract 1
- 239000010424 alunite Substances 0.000 abstract 1
- 229910052934 alunite Inorganic materials 0.000 abstract 1
- 239000001947 glycyrrhiza glabra rhizome/root Substances 0.000 abstract 1
- 239000010446 mirabilite Substances 0.000 abstract 1
- KPZTWMNLAFDTGF-UHFFFAOYSA-D trialuminum;potassium;hexahydroxide;disulfate Chemical compound [OH-].[OH-].[OH-].[OH-].[OH-].[OH-].[Al+3].[Al+3].[Al+3].[K+].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O KPZTWMNLAFDTGF-UHFFFAOYSA-D 0.000 abstract 1
- 229960004756 ethanol Drugs 0.000 description 20
- 238000000034 method Methods 0.000 description 11
- 239000002775 capsule Substances 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 6
- 239000013558 reference substance Substances 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- FCPVYOBCFFNJFS-LQDWTQKMSA-M benzylpenicillin sodium Chemical compound [Na+].N([C@H]1[C@H]2SC([C@@H](N2C1=O)C([O-])=O)(C)C)C(=O)CC1=CC=CC=C1 FCPVYOBCFFNJFS-LQDWTQKMSA-M 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229940126678 chinese medicines Drugs 0.000 description 1
- 201000001352 cholecystitis Diseases 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012738 dissolution medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000004900 laundering Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 229960002385 streptomycin sulfate Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a preparation method and an application of Danshilitong tablets, wherein the Danshilitong tablets are prepared by adopting 225 g of dichondra repens forst, 225 g of artemisiacapillaris thunb, 90 g of curcuma aromatica salisb, 150 g of tangerine peel, 45 g of scutellaria baicalensis georgi, 35 g of boswellia carteri, 80 g of mirabilite, 35 g of alunite, 45 g of rheum, 45 g of common burreed tuber, 45 g of cape jasmine, 25 g of licorice root, and 35 g of myrrh as bulk drugs. According to the prevent invention, ultra-fine crushing, supercritical fluid extraction, microwave extraction and macroporous resin absorption are performed to prepare the Danshilitong tablets, such that the hesperidin content is substantially increased. The present invention further provides the application of the Danshilitong tablets in preparation of mouse lymphoma cell YAC-1 proliferation inhibition drugs.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of preparation method of cholelithiasis treating Danshitongli tablet and suppress the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
Background technology
DanShiTongLi capsules standard No. WS-10324(ZD-0324)-2002, be recorded in national standard for traditional Chinese medicines compilation internal medicine liver and gall fascicle.By Herba Lysimachiae 225g, Herba Artemisiae Scopariae 225g, Radix Curcumae 90g, Pericarpium Citri Reticulatae 150g, Radix Scutellariae 45g, Olibanum 35g, Natrii Sulfas 80g, Alumen 35g, Radix Et Rhizoma Rhei 45g, Rhizoma Sparganii 45g, Fructus Gardeniae 45g, Radix Glycyrrhizae 25g, Myrrha 35g, as crude drug, made, there is clearing heat secreting bile, blood stasis dispelling calculus removing effect.For the acute and chronic cholecystitis due to dampness-heat in the liver and gallbladder, cholelithiasis etc.
In prior art, not yet there is DanShiTongLi capsules adopting micronizing, CO aspect extraction preparation
2the report that supercritical extraction, microwave technology and macroporous adsorption resin technology extract, and Chinese medicine is directly beaten the method that powder is got, decocting boils, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, is inconvenient to take, and has had a strong impact on this product and has applied clinically.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of preparation method of cholelithiasis treating Danshitongli tablet.
Another object of the present invention is to provide a kind of cholelithiasis treating Danshitongli tablet to suppress the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
The object of the invention is to realize by following scheme:
A kind of preparation method of cholelithiasis treating Danshitongli tablet, by Herba Lysimachiae 225g, Herba Artemisiae Scopariae 225g, Radix Curcumae 90g, Pericarpium Citri Reticulatae 150g, Radix Scutellariae 45g, Olibanum 35g, Natrii Sulfas 80g, Alumen 35g, Radix Et Rhizoma Rhei 45g, Rhizoma Sparganii 45g, Fructus Gardeniae 45g, Radix Glycyrrhizae 25g, Myrrha 35g, as crude drug, made, described preparation method is comprised of the following step: Natrii Sulfas, Alumen micronizing become the fine powder of particle size range 5-45 μ m standby, get Olibanum, Myrrha, be ground into fine powder, join CO
2in supercritical extraction 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 ℃, CO
2flow 1-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, standby, Olibanum after extraction, Myrrha residue and other nine tastes medical materials, pulverize, 70% ethanol that adds 3.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extract 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 70% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, by the Natrii Sulfas after micronizing, Alumen, supercritical extract, microwave extraction thing mixes and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, described CO
2in supercritical extraction, to account for the percent by volume of total extractant be 5% to entrainer.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, described CO
2in supercritical extraction, extracting pressure is 25MPa.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, described CO
2in supercritical extraction, extraction temperature is 60 ℃.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, described CO
2in supercritical extraction, extraction time is 200min.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, described microwave extracting power is 700W.
In the preparation method of above-mentioned cholelithiasis treating Danshitongli tablet, the each extraction time of described microwave extracting is 8 minutes.
Above-mentioned cholelithiasis treating Danshitongli tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
In prior art, DanShiTongLi capsules is oral, one time 5,3 times on the one.DanShiTongLi capsules dosage is large.The every heavy 0.3g of cholelithiasis treating Danshitongli tablet that adopts the inventive method to be prepared into only needs 2 at every turn, within 1st, takes 3 times.Greatly reduced dose having under the condition of more active component.This conclusion can be by following evidence.
The comparison of content of hesperidin in cholelithiasis treating Danshitongli tablet prepared by test one, distinct methods
1, instrument and reagent cholelithiasis treating Danshitongli tablet of the present invention: press embodiment 1 method preparation, use 1080g crude drug, make 500 through extracting, every heavy 0.3g.Former DanShiTongLi capsules, according to WS-10324(ZD-0324)-2002 standard method preparations, in contrast.Agilent1200 high performance liquid chromatograph; AE240 electronic analytical balance; Hesperidin reference substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
According to high performance liquid chromatography (appendix V D of Chinese Pharmacopoeia version in 2010), measure.
Strong to close silica gel be filler by octadecylsilane for chromatographic condition and system suitability; Methanol-water (40: 60) is mobile phase; Detection wavelength is 284nm; Number of theoretical plate calculates and should be not less than 3000 by Hesperidin peak.
It is appropriate that the preparation precision of reference substance solution takes Hesperidin reference substance, adds methanol and make every 1ml containing the solution of 38 μ g, obtains.
Cholelithiasis treating Danshitongli tablet of the present invention is got in the preparation of product need testing solution of the present invention, and porphyrize is got 0.06g,, accurately weighed, put in tool plug conical flask, precision adds Diluted Alcohol 25ml, weighed weight, close plug, supersound process (power 250w, frequency 30kHz) 40 minutes, lets cool, weighed weight, and with Diluted Alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate, obtain.
The DanShiTongLi capsules of this contrast is got in the preparation of reference substance need testing solution, mixes porphyrize, get 0.15g, accurately weighed, put in tool plug conical flask, precision adds Diluted Alcohol 25ml, weighed weight, close plug, supersound process (power 250w, frequency 30kHz) 40 minutes, lets cool, weighed weight, and with Diluted Alcohol, supply the weight of less loss, shake up, filter, get subsequent filtrate, obtain.
Algoscopy is accurate reference substance solution and each 10 μ l of need testing solution of drawing respectively, and injection liquid chromatography, measures, and obtains.
3, result
Result shows, in cholelithiasis treating Danshitongli tablet of the present invention, the content of Hesperidin is 6-10mg/ sheet; And the content of Hesperidin is 1.45mg/ grain in former DanShiTongLi capsules, doubly, in the situation that dose reduces, content of hesperidin improves a lot the 4-6 that every content of hesperidin is equivalent to original capsule content.
Above-mentioned research shows, the cholelithiasis treating Danshitongli tablet that adopts preparation method of the present invention to prepare, active constituent content is far away higher than WS-10324(ZD-0324) DanShiTongLi capsules prepared of the method recorded of-2002 standards.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is further described, so that those skilled in the art more understands the present invention, but this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following example, all technology realizing based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Herba Lysimachiae 225g, Herba Artemisiae Scopariae 225g, Radix Curcumae 90g, Pericarpium Citri Reticulatae 150g, Radix Scutellariae 45g, Olibanum 35g, Natrii Sulfas 80g, Alumen 35g, Radix Et Rhizoma Rhei 45g, Rhizoma Sparganii 45g, Fructus Gardeniae 45g, Radix Glycyrrhizae 25g, Myrrha 35g: become the fine powder of particle size range 5-45 μ m standby Natrii Sulfas, Alumen micronizing, get Olibanum, Myrrha, be ground into fine powder, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 25MPa, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 200min, obtains supercritical extract, standby, Olibanum after extraction, Myrrha residue and other nine tastes medical materials, pulverize, 70% ethanol that adds 3.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 700W, extract 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 70% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, by the Natrii Sulfas after micronizing, Alumen, supercritical extract, microwave extraction thing mixes and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
After testing, in finished product, the content of Hesperidin is 9.86mg/ sheet.
Embodiment 2
Get Herba Lysimachiae 225g, Herba Artemisiae Scopariae 225g, Radix Curcumae 90g, Pericarpium Citri Reticulatae 150g, Radix Scutellariae 45g, Olibanum 35g, Natrii Sulfas 80g, Alumen 35g, Radix Et Rhizoma Rhei 45g, Rhizoma Sparganii 45g, Fructus Gardeniae 45g, Radix Glycyrrhizae 25g, Myrrha 35g: become the fine powder of particle size range 5-45 μ m standby Natrii Sulfas, Alumen micronizing, get Olibanum, Myrrha, be ground into fine powder, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 30MPa, temperature 50 C, CO
2flow 3ml/g crude drug min, extraction time 180min, obtains supercritical extract, standby, Olibanum after extraction, Myrrha residue and other nine tastes medical materials, pulverize, 70% ethanol that adds 3.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600W, extract 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 70% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, by the Natrii Sulfas after micronizing, Alumen, supercritical extract, microwave extraction thing mixes and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
After testing, in finished product, the content of Hesperidin is 6.42mg/ sheet.
Embodiment 3
Get Herba Lysimachiae 225g, Herba Artemisiae Scopariae 225g, Radix Curcumae 90g, Pericarpium Citri Reticulatae 150g, Radix Scutellariae 45g, Olibanum 35g, Natrii Sulfas 80g, Alumen 35g, Radix Et Rhizoma Rhei 45g, Rhizoma Sparganii 45g, Fructus Gardeniae 45g, Radix Glycyrrhizae 25g, Myrrha 35g: become the fine powder of particle size range 5-45 μ m standby Natrii Sulfas, Alumen micronizing, get Olibanum, Myrrha, be ground into fine powder, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 15MPa, 30 ℃ of temperature, CO
2flow 1ml/g crude drug min, extraction time 220min, obtains supercritical extract, standby, Olibanum after extraction, Myrrha residue and other nine tastes medical materials, pulverize, 70% ethanol that adds 3.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 800W, extract 2 times, each 10 minutes, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 70% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, by the Natrii Sulfas after micronizing, Alumen, supercritical extract, microwave extraction thing mixes and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
After testing, in finished product, the content of Hesperidin is 8.26mg/ sheet.
Embodiment 4: cholelithiasis treating Danshitongli tablet suppresses the experimentation data of mouse lymphoma cell YAC-1 cell proliferation
1. experiment material
1.1 experiment cell strains
Mouse lymphoma cell YAC-1 cell, Shandong University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: cholelithiasis treating Danshitongli tablet of the present invention: press embodiment 1 method preparation.
Medicinal liquid liquid storage: take 100mg cholelithiasis treating Danshitongli tablet, be dissolved in 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ ldoff pipe subpackages ,-20 ℃ of storages, 0.2 μ m filter filters dehydrated alcohol in order to the use of matched group simultaneously.
1.3 experiment reagent
DMEM (the Cat.No.12100-061 Lot.No.758137 of GIBCO company); Hyclone (Lot.No.100419 of Tian Hang bio tech ltd, Zhejiang); NaHCO
3(the Cat.No.11810-033 Lot.No.1088387 of Shanghai Jiu Yi chemical reagent company limited); Trypsin(AMRESCO company); EDTA(AMRESCO company); Penicillin G Sodium Salt(AMRESCO company 1); Streptomycin Sulfate (AMRESCO); Dehydrated alcohol (Zibo Ya Dulan Trade Co., Ltd.); MTT (Biosharp lot number: 0793): the autogamy of PBS(laboratory);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DMIL); Visible-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRAMAX 190); CO
2incubator (FORMA model: 3111); (safe and sound company of Su Jing group manufactures model to super-clean bench: SW-CJ-ZFD); Pure water instrument (U.S. Sprlng company model: S/N 020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (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); 1cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2. experimental technique
1) YAC-1 cell uses DMEM+10%FBS in 37 ℃, 5%CO
2carry out cellar culture (10cm culture dish), when Growth of Cells is during to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 ℃ of digestion 2min, add wherein 5ml complete medium neutralization reaction, after piping and druming cell, proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjusts concentration of cell suspension 3 * 10
4individual/ml.
2) cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 ℃, 5%CO
2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add cholelithiasis treating Danshitongli tablet solution, continue to cultivate 24h.
4) after 24h, add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT), continue to cultivate 4h.
5) after 4h, buckle method is removed supernatant, with absorbent paper, pats dry gently, and low-speed oscillation 10min on shaking table, as entered 200 μ l dimethyl sulfoxide, is put in every hole, and crystal is fully dissolved.At enzyme-linked immunosorbent assay instrument 490nm place, measure the light absorption value in each hole.
6) background (do not add cell, only add culture fluid) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide), sets 6 multiple holes for every group.
7) result represents the suppression ratio of cell with medicine: cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value), control wells OD value * 100%.Experiment repeats 3 times.
3. statistical disposition
Adopt correlation analysis and Student t check in Microsoft Excel 2007 softwares, data represent with mean ± S.D..
4. experimental result
Statistical result showed after mtt assay experiment, with matched group comparison, when dosage reaches 5mg/ml, to YAC-1 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has utmost point significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 cholelithiasis treating Danshitongli tablet is on YAC-1 cell inhibitory effect impact research (X ± SD)
| Group | Drug level (mg/ml) | Suppression ratio (%) |
| Matched group | 0 | 0 |
| 1 | 5 | 8.42±7.03 |
| 2 | 10 | 15.98±8.69* |
| 3 | 15 | 27.03±10.77** |
| 4 | 20 | 36.56±12.64** |
Note: with matched group comparison, * P<O.01; * P<0.001.
5. experiment conclusion
Cholelithiasis treating Danshitongli tablet can suppress YAC-1 cell proliferation, reduces the Growth of Cells number of YAC-1 cell, and this effect is dose dependent.
Claims (9)
1. the preparation method of a cholelithiasis treating Danshitongli tablet, by Herba Lysimachiae 225g, Herba Artemisiae Scopariae 225g, Radix Curcumae 90g, Pericarpium Citri Reticulatae 150g, Radix Scutellariae 45g, Olibanum 35g, Natrii Sulfas 80g, Alumen 35g, Radix Et Rhizoma Rhei 45g, Rhizoma Sparganii 45g, Fructus Gardeniae 45g, Radix Glycyrrhizae 25g, Myrrha 35g, as crude drug, made, it is characterized in that, described preparation method is comprised of the following step: Natrii Sulfas, Alumen micronizing become the fine powder of particle size range 5-45 μ m standby, get Olibanum, Myrrha, be ground into fine powder, join CO
2in supercritical extraction 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 ℃, CO
2flow 1-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, standby, Olibanum after extraction, Myrrha residue and other nine tastes medical materials, pulverize, 70% ethanol that adds 3.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extract 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Diaion HP-10 macroporous adsorptive resins, 70% ethanol elution, collect 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, by the Natrii Sulfas after micronizing, Alumen, supercritical extract, microwave extraction thing mixes and adds starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.3g.
2. the preparation method of cholelithiasis treating Danshitongli tablet according to claim 1, is characterized in that, described CO
2in supercritical extraction, to account for the percent by volume of total extractant be 5% to entrainer.
3. the preparation method of cholelithiasis treating Danshitongli tablet according to claim 1, is characterized in that, described CO
2in supercritical extraction, extracting pressure is 25MPa.
4. the preparation method of cholelithiasis treating Danshitongli tablet according to claim 1, is characterized in that, described CO
2in supercritical extraction, extraction temperature is 60 ℃.
5. the preparation method of cholelithiasis treating Danshitongli tablet according to claim 1, is characterized in that, described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
6. according to the preparation method of cholelithiasis treating Danshitongli tablet claimed in claim 1, it is characterized in that described CO
2in supercritical extraction, extraction time is 200min.
7. the preparation method of cholelithiasis treating Danshitongli tablet according to claim 1, is characterized in that, described microwave extracting power is 700W.
8. the preparation method of cholelithiasis treating Danshitongli tablet according to claim 1, is characterized in that, the each extraction time of described microwave extracting is 8 minutes.
9. cholelithiasis treating Danshitongli tablet claimed in claim 1 suppresses the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310672109.5A CN103751701B (en) | 2013-12-10 | 2013-12-10 | A kind of preparation method of cholelithiasis treating Danshitongli tablet and the application in preparation suppression lymphoma cell YAC-1 cell proliferation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310672109.5A CN103751701B (en) | 2013-12-10 | 2013-12-10 | A kind of preparation method of cholelithiasis treating Danshitongli tablet and the application in preparation suppression lymphoma cell YAC-1 cell proliferation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103751701A true CN103751701A (en) | 2014-04-30 |
| CN103751701B CN103751701B (en) | 2016-03-02 |
Family
ID=50519128
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310672109.5A Expired - Fee Related CN103751701B (en) | 2013-12-10 | 2013-12-10 | A kind of preparation method of cholelithiasis treating Danshitongli tablet and the application in preparation suppression lymphoma cell YAC-1 cell proliferation thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103751701B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102988723A (en) * | 2012-10-08 | 2013-03-27 | 李正梅 | Preparation method and application of yin tonifying and calming tablet |
-
2013
- 2013-12-10 CN CN201310672109.5A patent/CN103751701B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102988723A (en) * | 2012-10-08 | 2013-03-27 | 李正梅 | Preparation method and application of yin tonifying and calming tablet |
Non-Patent Citations (4)
| Title |
|---|
| 俞丽霞等: "《中药药理学》", 31 December 2012, 浙江大学出版社 * |
| 国家药品监督管理局: "胆石通利胶囊", 《国家药品标准（试行）颁布件》 * |
| 赵余庆: "《中药及天然产物提取制备关键技术》", 31 January 2012, 中国医药科技出版社 * |
| 黄红兵: "《抗肿瘤中药临床应用与图谱》", 31 January 2008, 广东科技出版社 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103751701B (en) | 2016-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103566023A (en) | Preparation method and application of toothache anti-inflammation tablets | |
| CN103655843A (en) | Yikangbuyuan tablet, and preparation method and application thereof | |
| CN102988489B (en) | Preparation method and application of nose comforting tablet | |
| CN103028005B (en) | Preparation method of anti-lumbago tablet and application thereof | |
| CN103656493B (en) | A kind of preparation method of blood stasis dispelling stomach reinforcing sheet and application | |
| CN102988550B (en) | Preparation method and application of qinlian tablet | |
| CN103800818A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting myeloma cell FO from cell proliferation | |
| CN103800816A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting stomach cancer cell MFC from cell proliferation | |
| CN102836381A (en) | Preparation method and application of middle calming tablet | |
| CN102988611B (en) | Preparation method and application of antihypertension tablet | |
| CN103784872B (en) | A kind of preparation method of cholelithiasis treating Danshitongli tablet and the application in preparation suppression Ascitic Tumor Cells SAC-II C3 cell proliferation thereof | |
| CN103751700A (en) | Danshilitong tablet preparation method and application of Danshilitong tablets in preparation of lymphoid neoplasm cell P388D1 proliferation inhibition drugs | |
| CN103751698A (en) | Danshilitong tablet preparation method and application of Danshilitong tablets in preparation of prostatic cancer cell RM-1 proliferation inhibition drugs | |
| CN103768514A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting myeloma cell SP2/0 from cell proliferation | |
| CN103768517A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting mouse lung cancer cell LLC from cell proliferation | |
| CN103690647A (en) | Preparation method of Yikangbuyuan tablet and application of Yikangbuyuan tablet in medicines for inhibiting mouse leukaemia cell L6565 proliferation | |
| CN103768516A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting stomach cancer cell MFC from cell proliferation | |
| CN103751424A (en) | Preparation method of cold tablet containing Artemisia carvifolia and holly root, and application of cold tablet in teratoma cell F9 cell proliferation inhibition medicines. | |
| CN103751701B (en) | A kind of preparation method of cholelithiasis treating Danshitongli tablet and the application in preparation suppression lymphoma cell YAC-1 cell proliferation thereof | |
| CN103751699A (en) | Danshilitong tablet preparation method and application of Danshilitong tablets in preparation of breast tumor cell C127 proliferation inhibition drugs | |
| CN103768513B (en) | A kind of preparation method of cholelithiasis treating Danshitongli tablet and the application in preparation inhibition teratocarcinoma cell P19 cell proliferation medicine thereof | |
| CN103768509A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting Leydig's cell tumor cell MLTC-1 from cell proliferation | |
| CN103768508A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting leukaemia cell P815 from cell proliferation | |
| CN103768512A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting lymphomata cell EL4.IL-2 from cell proliferation | |
| CN103800819A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting lung cancer cell Hepa1-6 from cell proliferation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| CB03 | Change of inventor or designer information |
Inventor after: Tang Xiaochun Inventor after: Liu Shuzhen Inventor after: Hou Ling Inventor after: Liu Zenghua Inventor after: Li Yan Inventor before: Sun Bo |
|
| COR | Change of bibliographic data | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20160201 Address after: 266500 Huangdao District Hospital of traditional Chinese medicine, Hainan Island Road 158, Huangdao District, Shandong, Qingdao, Qingdao Applicant after: Tang Xiaochun Address before: 250001 Shandong Province, Ji'nan City District Court building 2303 No. 288 Applicant before: JINAN XINQIDIAN MEDICINE SCIENCE & TECHNOLOGY CO., LTD. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160302 Termination date: 20161210 |