CN103800498A - Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting mouse prostate cancer cell RM-1 from cell proliferation - Google Patents
Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting mouse prostate cancer cell RM-1 from cell proliferation Download PDFInfo
- Publication number
- CN103800498A CN103800498A CN201310658881.1A CN201310658881A CN103800498A CN 103800498 A CN103800498 A CN 103800498A CN 201310658881 A CN201310658881 A CN 201310658881A CN 103800498 A CN103800498 A CN 103800498A
- Authority
- CN
- China
- Prior art keywords
- preparation
- extraction
- cold tablet
- common cold
- armeniaca mume
- 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 35
- 239000003814 drug Substances 0.000 title claims abstract description 23
- 229940079593 drug Drugs 0.000 title claims abstract description 15
- 230000004663 cell proliferation Effects 0.000 title claims abstract description 9
- 230000002401 inhibitory effect Effects 0.000 title abstract description 4
- 206010060862 Prostate cancer Diseases 0.000 title abstract 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 title abstract 2
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 16
- 238000000874 microwave-assisted extraction Methods 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 53
- 235000011158 Prunus mume Nutrition 0.000 claims description 42
- 244000018795 Prunus mume Species 0.000 claims description 42
- 201000009240 nasopharyngitis Diseases 0.000 claims description 34
- 238000000605 extraction Methods 0.000 claims description 22
- 239000000284 extract Substances 0.000 claims description 16
- 239000008187 granular material Substances 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 9
- 208000009956 adenocarcinoma Diseases 0.000 claims description 7
- 241000628997 Flos Species 0.000 claims description 6
- 241000699670 Mus sp. Species 0.000 claims description 6
- 229940126678 chinese medicines Drugs 0.000 claims description 6
- 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
- 238000004064 recycling Methods 0.000 claims description 5
- 229920005989 resin Polymers 0.000 claims description 5
- 239000011347 resin Substances 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 239000008107 starch Substances 0.000 claims description 5
- VTAJIXDZFCRWBR-UHFFFAOYSA-N Licoricesaponin B2 Natural products C1C(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2)C(O)=O)C)(C)CC2)(C)C2C(C)(C)CC1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O VTAJIXDZFCRWBR-UHFFFAOYSA-N 0.000 abstract description 11
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 abstract description 11
- 229960004949 glycyrrhizic acid Drugs 0.000 abstract description 11
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 abstract description 11
- 239000001685 glycyrrhizic acid Substances 0.000 abstract description 11
- 235000019410 glycyrrhizin Nutrition 0.000 abstract description 11
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 abstract description 11
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 2
- 235000015700 Artemisia abrotanum Nutrition 0.000 abstract 1
- 244000249062 Artemisia abrotanum Species 0.000 abstract 1
- 235000002555 Citrus medica var sarcodactylis Nutrition 0.000 abstract 1
- 240000007126 Citrus medica var. sarcodactylis Species 0.000 abstract 1
- 241001290198 Dicliptera Species 0.000 abstract 1
- 241000131458 Elsholtzia Species 0.000 abstract 1
- 240000004670 Glycyrrhiza echinata Species 0.000 abstract 1
- 235000001453 Glycyrrhiza echinata Nutrition 0.000 abstract 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 abstract 1
- 235000017382 Glycyrrhiza lepidota Nutrition 0.000 abstract 1
- 241000209035 Ilex Species 0.000 abstract 1
- 235000003332 Ilex aquifolium Nutrition 0.000 abstract 1
- 235000002296 Ilex sandwicensis Nutrition 0.000 abstract 1
- 235000002294 Ilex volkensiana Nutrition 0.000 abstract 1
- 235000011755 Nepeta hederacea Nutrition 0.000 abstract 1
- 244000215554 Nepeta hederacea Species 0.000 abstract 1
- 239000008186 active pharmaceutical agent Substances 0.000 abstract 1
- 229940010454 licorice Drugs 0.000 abstract 1
- 229960004756 ethanol Drugs 0.000 description 20
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 238000000034 method Methods 0.000 description 11
- 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
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 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
- 239000006228 supernatant Substances 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
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 235000019640 taste Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- 206010011224 Cough Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 241000167880 Hirundinidae Species 0.000 description 1
- ILRKKHJEINIICQ-OOFFSTKBSA-N Monoammonium glycyrrhizinate Chemical compound N.O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@H]1CC[C@]2(C)[C@H]3C(=O)C=C4[C@@H]5C[C@](C)(CC[C@@]5(CC[C@@]4(C)[C@]3(C)CC[C@H]2C1(C)C)C)C(O)=O)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O ILRKKHJEINIICQ-OOFFSTKBSA-N 0.000 description 1
- 206010028748 Nasal obstruction Diseases 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
- 229960000583 acetic acid Drugs 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 230000001754 anti-pyretic effect 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
- 235000019658 bitter taste Nutrition 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 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
- 230000000694 effects Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 231100000869 headache Toxicity 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
- 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
- 239000003507 refrigerant Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 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
- 238000009923 sugaring Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of traditional Chinese medicines, and particularly provides a preparation method and application of a Qingmei cold tablet. The preparation method comprises the following steps: by using 200g of southernwood, 267g of finger citron, 134g of elsholtzia, 200g of dicliptera, 134g of licorice, 134g of cat's-foot and 267g of holly root as active pharmaceutical ingredients, carrying out supercritical extraction and microwave extraction so that the glycyrrhizic acid content is greatly enhanced. The invention also provides application of the Qingmei cold tablet in preparing drugs for inhibiting mouse prostate cancer cell RM-1 from cell proliferation.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in inhibition mice row adenocarcinoma cell RM-1 cell proliferation medicine thereof.
Background technology
Armeniaca mume Sieb. cold granules standard No. WS-11318 (ZD-1318)-2002, is recorded in national standard for traditional Chinese medicines compilation internal medicine taste fascicle.Made as crude drug by Herba Artemisiae Annuae 200g, Folium vilicis Negundo 267g, Herba Moslae 134g, Herba Diclipterae Chinensis 200g, Radix Glycyrrhizae 134g, Herba Centellae 134g, Flos Ilicis Asprellae 267g, there is refrigerant antipyretic effect.For anemopyretic cold, headache, cough, nasal obstruction.
In prior art, not yet have Armeniaca mume Sieb. cold granules at the report that adopts supercritical and microwave technology aspect extraction preparation, and volatile oil extracts, the method that decocting boils, technique is coarse, backward, and impurity is many, cause patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of preparation method of Armeniaca mume Sieb. common cold tablet.
Another object of the present invention is to provide a kind of Armeniaca mume Sieb. common cold tablet to suppress the application in mice row adenocarcinoma cell RM-1 cell proliferation medicine in preparation.
The object of the invention is to realize by following scheme:
A kind of preparation method of Armeniaca mume Sieb. common cold tablet, made as crude drug by Herba Artemisiae Annuae 200g, Folium vilicis Negundo 267g, Herba Moslae 134g, Herba Diclipterae Chinensis 200g, Radix Glycyrrhizae 134g, Herba Centellae 134g, Flos Ilicis Asprellae 267g, described preparation method is made up of the following step: get Herba Artemisiae Annuae, Herba Moslae, 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 l-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 50% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extracts 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 150, every heavy 0.3g.
In the preparation method of above-mentioned Armeniaca mume Sieb. common cold 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 Armeniaca mume Sieb. common cold tablet, described CO
2in supercritical extraction, extracting pressure is 25MPa.
In the preparation method of above-mentioned Armeniaca mume Sieb. common cold tablet, described CO
2in supercritical extraction, extraction temperature is 60 ℃.
In the preparation method of above-mentioned Armeniaca mume Sieb. common cold tablet, described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
In the preparation method of above-mentioned Armeniaca mume Sieb. common cold tablet, described CO
2in supercritical extraction, extraction time is 200min.
In the preparation method of above-mentioned Armeniaca mume Sieb. common cold tablet, described microwave extracting power is 700W.
In the preparation method of above-mentioned Armeniaca mume Sieb. common cold tablet, the each extraction time of described microwave extracting is 8 minutes.
Above-mentioned Armeniaca mume Sieb. common cold tablet suppresses the application in mice row adenocarcinoma cell RM-1 cell proliferation medicine in preparation.
In prior art, each 1 bag of Armeniaca mume Sieb. cold granules, every bag of 15g, 2-3 time on the one.Armeniaca mume Sieb. cold granules dosage is large, and granule preparation need to add a large amount of sugar, and diabetics is not suitable for, and the words taste of not sugaring is not good, and patient is not easy under clothes, and compliance is poor.The every heavy 0.3g of Armeniaca mume Sieb. common cold tablet that adopts the inventive method to be prepared into only needs 2 at every turn, within 1st, takes 2-3 time.Under the condition with more active component, greatly reduce dose.This conclusion can be by following evidence.And be prepared into tablet, patient swallows with water, dosage is little and without bitterness, patient is easy to accept.
The comparison of glycyrrhizic acid content in Armeniaca mume Sieb. common cold tablet prepared by test one, distinct methods
L, instrument and reagent Armeniaca mume Sieb. common cold tablet of the present invention: press embodiment 1 method preparation, use 1510g crude drug, make 150 through extracting, every heavy 0.3g.Former Armeniaca mume Sieb. cold granules, prepares according to WS-11318 (ZD-1318)-2002 standard method.Agilent1200 high performance liquid chromatograph; METTLER AE240 electronic analytical balance; Glycyrrhizic acid reference substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
Measure according to high performance liquid chromatography (appendix VI D of Chinese Pharmacopoeia version in 2010).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler; Methanol-0.2mol/L Spirit of Mindererus .-glacial acetic acid (67: 33: 1) is mobile phase; Detection wavelength is 250nm.Number of theoretical plate calculates and should be not less than 3000 by glycyrrhizic acid peak.
It is appropriate that the preparation precision of reference substance solution takes monoammonium glycyrrhizinate reference substance, adds 90% methanol and make the solution of every 1ml containing 0.4mg, to obtain final product.
Armeniaca mume Sieb. common cold tablet of the present invention is got in the preparation of need testing solution, and porphyrize, gets 0.08g, accurately weighed, to put in tool plug conical flask, precision adds 90% methanol 10ml, close plug, weighed weight, supersound process 30 minutes, let cool, more weighed weight, supply the weight of less loss with 90% methanol, centrifugal, (0.45 μ m) filters microporous filter membrane for supernatant, gets subsequent filtrate, to obtain final product.
The Armeniaca mume Sieb. cold granules of this contrast is got in the preparation of reference product need testing solution, and porphyrize, mixes, get 2g, accurately weighed, put in tool plug conical flask, precision adds 90% methanol 10ml, close plug, weighed weight, supersound process 30 minutes, lets cool, more weighed weight, supply the weight of less loss with 90% methanol, centrifugal, (0.45 μ m) filters microporous filter membrane for supernatant, get subsequent filtrate, to obtain final product.
Algoscopy is accurate reference substance solution and the each 20 μ l of need testing solution of drawing respectively, and injection liquid chromatography, measures, and to obtain final product.3, result
Result shows, in Armeniaca mume Sieb. common cold tablet of the present invention, the content of glycyrrhizic acid is 35-70mg/ sheet; And the content of glycyrrhizic acid is 14.5mg/ bag in former Armeniaca mume Sieb. cold granules, be 5-10 times of former granule content each serving the glycyrrhizic acid content of 2 of consumptions, in the situation that dose reduces, glycyrrhizic acid content improves a lot.
Above-mentioned research shows, the Armeniaca mume Sieb. common cold tablet that adopts preparation method of the present invention to prepare, Armeniaca mume Sieb. cold granules prepared by the method that active constituent content is recorded higher than WS-11318 (ZD-1318)-2002 standard far away.
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 Artemisiae Annuae 200g, Folium vilicis Negundo 267g, Herba Moslae 134g, Herba Diclipterae Chinensis 200g, Radix Glycyrrhizae 134g, Herba Centellae 134g, Flos Ilicis Asprellae 267g, Herba Artemisiae Annuae, Herba Moslae are joined to 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, for subsequent use; Get all the other Chinese medicines, pulverize, add 50% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 700W, extracts 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 150, every heavy 0.3g.
After testing, in finished product, the content of glycyrrhizic acid is 69.8mg/ sheet.
Embodiment 2
Get Herba Artemisiae Annuae 200g, Folium vilicis Negundo 267g, Herba Moslae 134g, Herba Diclipterae Chinensis 200g, Radix Glycyrrhizae 134g, Herba Centellae 134g, Flos Ilicis Asprellae 267g, Herba Artemisiae Annuae, Herba Moslae are joined to 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, for subsequent use; Get all the other Chinese medicines, pulverize, add 50% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600W, extracts 2 times, each 5 minutes, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 150, every heavy 0.3g.
After testing, in finished product, the content of glycyrrhizic acid is 56.2mg/ sheet.
Embodiment 3
Get Herba Artemisiae Annuae 200g, Folium vilicis Negundo 267g, Herba Moslae 134g, Herba Diclipterae Chinensis 200g, Radix Glycyrrhizae 134g, Herba Centellae 134g, Flos Ilicis Asprellae 267g, Herba Artemisiae Annuae, Herba Moslae are joined to 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, for subsequent use; Get all the other Chinese medicines, pulverize, add 50% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 800W, extracts 2 times, each 10 minutes, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 150, every heavy 0.3g.
After testing, in finished product, the content of glycyrrhizic acid is 36.9mg/ sheet.
Embodiment 4: Armeniaca mume Sieb. common cold tablet suppresses the experimentation data of mice row adenocarcinoma cell RM-1 cell proliferation
1. experiment material
1.1 experiment cell strains
Mice row adenocarcinoma cell RM-1 cell, Shandong University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: Armeniaca mume Sieb. common cold tablet of the present invention: press embodiment 1 method preparation.
Medicinal liquid liquid storage: take 100mg Armeniaca mume Sieb. common cold 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 the use of dehydrated alcohol in order to 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); NaHC0
3(Shanghai hundred million Cat.No.11810-033Lot.No.1088387 of chemical reagent company limited of a specified duration); 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): PBS(laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DMIL); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX 190); C0
2incubator (FORMA model: 3111); Super-clean bench (safe and sound company of Su Jing group manufactures model: SW-CJ-ZFD); Pure water instrument (Sprlng company of U.S. 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) RM-1 cell uses DMEM+10%FBS in 37 ℃, 5%C0
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%C0
2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add Armeniaca mume Sieb. common cold 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, pats dry gently with absorbent paper, 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.Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place.
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, in the time that dosage reaches 5mg/ml, to RM-1 cell inhibitory effect variant (P<0.05), dosage this difference in the time of 10mg/ml has significance (P<0.01), has utmost point significant difference (P<0.001) in the time that dosage reaches 15-20mg/ml.
Table 1 Armeniaca mume Sieb. common cold tablet is on RM-1 cell inhibitory effect impact research (X ± SD)
| Group | Drug level (mg/ml) | Suppression ratio (%) |
| Matched group | 0 | 0 |
| 1 | 5 | 9.99±7.52 |
| 2 | 10 | 18.21±12.12* |
| 3 | 15 | 34.12±17.30** |
| 4 | 20 | 45.39±16.20** |
Note: with matched group comparison, * P<O.01; * P<0.001.
5. experiment conclusion
Armeniaca mume Sieb. common cold tablet of the present invention can suppress RM-1 cell proliferation, reduces the Growth of Cells number of RM-1 cell, and this effect is dose dependent.
Claims (9)
1. the preparation method of an Armeniaca mume Sieb. common cold tablet, made as crude drug by Herba Artemisiae Annuae 200g, Folium vilicis Negundo 267g, Herba Moslae 134g, Herba Diclipterae Chinensis 200g, Radix Glycyrrhizae 134g, Herba Centellae 134g, Flos Ilicis Asprellae 267g, it is characterized in that, described preparation method is made up of the following step: get Herba Artemisiae Annuae, Herba Moslae, 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 l-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 50% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extracts 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 150, every heavy 0.3g.
2. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that described CO
2in supercritical extraction, to account for the percent by volume of total extractant be 5% to entrainer.
3. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that described CO
2in supercritical extraction, extracting pressure is 25MPa.
4. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that described CO
2in supercritical extraction, extraction temperature is 60 ℃.
5. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
6. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that described CO
2in supercritical extraction, extraction time is 200min.
7. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that, described microwave extracting power is 700W.
8. according to the preparation method of the Armeniaca mume Sieb. common cold tablet described in claim l, it is characterized in that, the each extraction time of described microwave extracting is 8 minutes.
9. the Armeniaca mume Sieb. common cold tablet described in claim l suppresses the application in mice row adenocarcinoma cell RM-1 cell proliferation medicine in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310658881.1A CN103800498B (en) | 2013-12-06 | 2013-12-06 | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression row adenocarcinoma cell RM-1 cell proliferation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310658881.1A CN103800498B (en) | 2013-12-06 | 2013-12-06 | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression row adenocarcinoma cell RM-1 cell proliferation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103800498A true CN103800498A (en) | 2014-05-21 |
| CN103800498B CN103800498B (en) | 2016-01-20 |
Family
ID=50698193
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310658881.1A Expired - Fee Related CN103800498B (en) | 2013-12-06 | 2013-12-06 | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression row adenocarcinoma cell RM-1 cell proliferation thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103800498B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104161852A (en) * | 2014-07-31 | 2014-11-26 | 济南爱思医药科技有限公司 | Preparation method for nine-composition hemorrhoid tablet and application thereof to medicines inhibiting cell proliferation of prostate cancer cell RM-1 |
| CN104288656A (en) * | 2014-10-16 | 2015-01-21 | 济南新起点医药科技有限公司 | Preparation method of fomes officinalis cough and asthma treating tablets and application of fomes officinalis cough and asthma treating tablets in preparation of medicine for inhibiting cell proliferation of mouse prostatic cancer cell RM-1 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102973667A (en) * | 2012-10-15 | 2013-03-20 | 李正梅 | Preparation method and application of cold-fever tablet |
-
2013
- 2013-12-06 CN CN201310658881.1A patent/CN103800498B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102973667A (en) * | 2012-10-15 | 2013-03-20 | 李正梅 | Preparation method and application of cold-fever tablet |
Non-Patent Citations (5)
| Title |
|---|
| 周岱瀚: "《中医肿瘤学》", 30 June 2011, article "青蒿", pages: 463-464 * |
| 国家药品监督管理局: "《国家药品标准（试行）颁布件》", 16 November 2002, article "青梅感冒颗粒", pages: 339-342 * |
| 梅全喜: "《广东地产药材研究》", 31 May 2011, article "佛手，岗梅", pages: 346-347 - 364-366 * |
| 程剑华等: "《抗癌植物药与其验方》", 30 April 1998, article "狗肝菜", pages: 475-476 * |
| 范文昌等: "《广东地产清热解毒药物大全》", 31 July 2011, article "积雪草", pages: 522 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104161852A (en) * | 2014-07-31 | 2014-11-26 | 济南爱思医药科技有限公司 | Preparation method for nine-composition hemorrhoid tablet and application thereof to medicines inhibiting cell proliferation of prostate cancer cell RM-1 |
| CN104288656A (en) * | 2014-10-16 | 2015-01-21 | 济南新起点医药科技有限公司 | Preparation method of fomes officinalis cough and asthma treating tablets and application of fomes officinalis cough and asthma treating tablets in preparation of medicine for inhibiting cell proliferation of mouse prostatic cancer cell RM-1 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103800498B (en) | 2016-01-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103655843A (en) | Yikangbuyuan tablet, and preparation method and application thereof | |
| CN103800498B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression row adenocarcinoma cell RM-1 cell proliferation thereof | |
| CN103768211B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression lymphoma cell EL4 cell proliferation thereof | |
| CN103768266A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting mouse leukaemia cell L6565 from cell proliferation | |
| CN103800499A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting melanoma cell B16 from cell proliferation | |
| CN103768262A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting myeloma cell FO from cell proliferation | |
| CN103768267A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting lung cancer cell H22 from cell proliferation | |
| CN103768263A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting cerebral nerve oncocyte Neuro-2a 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. | |
| CN103893411A (en) | Preparation method and application of green plum cold tablets | |
| CN103751350B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression leukaemia L1210 cell proliferation thereof | |
| CN103735960A (en) | Application of boenninghausenia sessilicarpa cough-asthma tablet to prepare medicaments for inhibiting lung cancer cell LLC proliferation | |
| CN103768207B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression hepatoma carcinoma cell Hepa1-6 cell proliferation thereof | |
| CN103690647A (en) | Preparation method of Yikangbuyuan tablet and application of Yikangbuyuan tablet in medicines for inhibiting mouse leukaemia cell L6565 proliferation | |
| CN103768265A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting lung cancer cell LLC from cell proliferation | |
| CN103768212B (en) | The preparation method of a kind of Armeniaca mume Sieb. common cold tablet and the application in suppressing stomach cancer cell MFC cell proliferation thereof | |
| CN103800569A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting lymphoma cell YAC-1 from cell proliferation | |
| CN103768264A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting teratoma cell P19 from cell proliferation | |
| CN103655841B (en) | Preparation method of Yikangbuyuan tablet and application of Yikangbuyuan tablet in medicines for inhibiting mouse lymphoid cancer cell P388D1 proliferation | |
| CN103655839B (en) | Preparation method of Yikangbuyuan tablet and application of Yikangbuyuan tablet in medicines for inhibiting mouse lung cancer cell LLC proliferation | |
| CN103751425A (en) | Preparation method of Qingmei cold-treatment tablet and use of Qingmei cold-treatment tablet in drug for inhibiting breast tumor cell C127 proliferation | |
| CN103751351B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression leydig cell tumor of testis cell MLTC-1 cell proliferation thereof | |
| CN103768210B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression anus sarcoma cell S-180 cell proliferation thereof | |
| CN103768209B (en) | A kind of preparation method of Armeniaca mume Sieb. common cold tablet and the application in suppression myeloma cell P3X63AG8 cell proliferation thereof | |
| CN103800770A (en) | Application of boenninghausenia cough tablet in preparing medicine for inhibiting cell proliferation of breast tumor cell C127 |
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: Bao Ming Inventor after: Hou Xuanxuan Inventor after: Wang Ran Inventor before: Sun Bo |
|
| COR | Change of bibliographic data | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20151223 Address after: 510515 Guangzhou, Baiyun District, Guangzhou, North Road, No., No. 1838 Applicant after: NANFANG HOSPITAL, SOUTHERN MEDICAL University Address before: 250001 Shandong Province, Ji'nan City District Court building 2303 No. 288 Applicant before: JINAN XINQIDIAN MEDICAL 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: 20160120 |