CN103494930A - Preparation method and application of compound houttuynia cordata tablets - Google Patents
Preparation method and application of compound houttuynia cordata tablets Download PDFInfo
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- CN103494930A CN103494930A CN201310466159.8A CN201310466159A CN103494930A CN 103494930 A CN103494930 A CN 103494930A CN 201310466159 A CN201310466159 A CN 201310466159A CN 103494930 A CN103494930 A CN 103494930A
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Abstract
The invention provides a preparation method of compound houttuynia cordata tablets. The compound houttuynia cordata tablets are prepared from 583g of houttuynia cordata, 150g of scutellaria baicalensis, 150g of isatis root, 58g of fructus forsythiae and 58g of honeysuckle which are used as raw medicinal materials by adopting a supercritical extraction method, so that the content is greatly increased and the intake dosage is reduced. The invention also provides an application of the compound houttuynia cordata tablets in preparation of a medicament for restricting cell proliferation of rat hepatoma carcinoma cells RH-35.
Description
Technical field
The present invention relates to the Chinese medicine preparation technical field, be specifically related to a kind of preparation method and application of FUFANG YUXINGCAO PIAN.
Background technology
The FUFANG YUXINGCAO PIAN pharmacopeia, write out a prescription as Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, and the above five tastes, get Herba Houttuyniae 200g, be broken into fine powder with Fructus Forsythiae, Flos Lonicerae powder, remaining Herba Houttuyniae and Radix Scutellariae, Radix Isatidis decoct with water secondary, each 2 hours, collecting decoction, filter, and filtrate is condensed into thick paste, add above-mentioned fine powder, mix, drying, be ground into fine powder, granulation, drying, be pressed into 1000, sugar coating, obtain.Can heat-clearing and toxic substances removing.What for affection due to external wind and heat, cause has sore throat; Acute pharyngitis, tonsillitis have the wind heat patient.
In prior art, not yet there is FUFANG YUXINGCAO PIAN to adopt the report of supercritical technology extracting aspect preparation, and adopt the method that powder and decocting boil of beating, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied 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 of FUFANG YUXINGCAO PIAN.
Another object of the present invention is to provide a kind of FUFANG YUXINGCAO PIAN to suppress the application in rat hepatoma cell RH-35 cell proliferation medicine in preparation.
Technical scheme: the objective of the invention is to realize by following scheme:
A kind of preparation method of FUFANG YUXINGCAO PIAN, by Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, as crude drug, made, described method is comprised of the following step: get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, join in CO2 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 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 500, every heavy 0.5g.
The preparation method of above-mentioned a kind of FUFANG YUXINGCAO PIAN, described CO
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned a kind of FUFANG YUXINGCAO PIAN, described CO
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of FUFANG YUXINGCAO PIAN suppresses the application in rat hepatoma cell RH-35 cell proliferation medicine in preparation, FUFANG YUXINGCAO PIAN is made as crude drug by Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, preparation method is comprised of the following step: get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, join CO
2in the 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 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1500, every heavy 0.5g.
Above-mentioned FUFANG YUXINGCAO PIAN suppresses the application in rat hepatoma cell RH-35 cell proliferation medicine, CO described in the compound houttuynin piece preparation method in preparation
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned FUFANG YUXINGCAO PIAN suppresses the application in rat hepatoma cell RH-35 cell proliferation medicine, CO described in the compound houttuynin piece preparation method in preparation
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
In prior art, every 0.5g of FUFANG YUXINGCAO PIAN, each 6,3 times on the one, the every 0.5g of FUFANG YUXINGCAO PIAN that adopts the present invention to be prepared into, but the medical material amount contained is original 2 times, therefore only needs 3 at every turn, within 1st, take 3 times, greatly reduced dose having under the condition of more active component.
The specific embodiment
Form by the following examples, foregoing of the present invention is described in further detail again, 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 realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g joins in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, 30 ℃ of temperature, CO
2flow 1m1/g crude drug min, extraction time 150min, obtain supercritical extract,, add starch, 70% ethanol granule processed, drying, tabletting, make 500, every heavy 0.5g.
Embodiment 2
Get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g joins CO
2in the supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa, temperature 50 C, CO
2flow 3m1/g crude drug min, extraction time 180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 500, every heavy 0.5g.
Embodiment 3
Get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g joins CO
2in the supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, 40 ℃ of temperature, CO
2flow 2m1/g crude drug min, extraction time 160min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 500, every heavy 0.5g.
Embodiment 4: FUFANG YUXINGCAO PIAN suppresses the experimentation data of RH-35 cell proliferation
1 experiment material
1.1 experiment cell strain
Rat hepatoma cell (RH-35), Nanjing Zheng Liang Pharmaceutical Technology Co., Ltd laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: FUFANG YUXINGCAO PIAN of the present invention: press embodiment 3 method preparations.
The medicinal liquid liquid storage: take the 100mg FUFANG YUXINGCAO PIAN, be dissolved in the 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ l doff pipe packing, and-20 ℃ of storages, 0.2 μ m filter filters the use of dehydrated alcohol in order to matched group simultaneously.
1.3 experiment reagent
The Cat.No.12100-061Lot.No.758137 of DMEM(GIBCO company); Hyclone (Hangzhoupro, sky, Zhejiang bio tech ltd Lot.No.100419); NaHCO3(Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.No.1088387 of a specified duration); Trypsin(AMRESCO company lot number: 2010/04); EDTA(AMRESCO company lot number: 2009/10); Penicillin G Sodium Salt(AMRESCO company lot number: 2010242); Streptomycin Sulfate(AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); The autogamy of PBS(laboratory);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRA MAX190); CO2 incubator (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. Spring company model: S/N020579); 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); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques
1) the RH-35 cell carries out cellar culture (10cm culture dish) with DMEM+10%FBS in 37 ℃, 5%CO2, when Growth of Cells during to logarithmic (log) phase, collecting cell, discard 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 the piping and druming cell, it is proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjust 3 * 104/ml of concentration of cell suspension.
2) the 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%CO2) cellar culture.
3) according to the Growth of Cells situation, generally grow to 50%-70%, add FUFANG YUXINGCAO PIAN solution, continue to cultivate 24h.
4) add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.
5) after 4h, the buckle method is removed supernatant, with absorbent paper, pats dry gently, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, 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), set 6 multiple holes for every group.
7) with medicine, the suppression ratio to cell means result:
Cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value * 100%.Experiment repeats 3 times.
3 statistical dispositions
Adopt correlation analysis and Student t check in Microsoft Excel2003 software, data mean with mean ± S.D..
4 experimental results
Statistical result showed after the mtt assay experiment, with matched group, compare, when dosage reaches 5mg/ml, to RH-35 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), and utmost point significant difference (P<0.001) is arranged when dosage reaches 15-20mg/ml.
Table 1 FUFANG YUXINGCAO PIAN is on RH-35 cell inhibitory effect impact research (X ± SD)
Annotate: compare * P<0.01 with matched group; * P<0.001
5 experiment conclusion
FUFANG YUXINGCAO PIAN can suppress the RH-35 cell proliferation, reduces the Growth of Cells number of RH-35 cell, and this effect is dose dependent.
Claims (6)
1. the preparation method of a FUFANG YUXINGCAO PIAN, by Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, as crude drug, made, it is characterized in that described method is comprised of the following step: get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, join in CO2 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 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1500, every heavy 0.5g.
2. a kind of preparation method of FUFANG YUXINGCAO PIAN according to claim 1, is characterized in that described CO
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of preparation method of FUFANG YUXINGCAO PIAN according to claim 1, is characterized in that described CO
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
4. a kind of FUFANG YUXINGCAO PIAN suppresses the application in rat hepatoma cell RH-35 cell proliferation medicine in preparation according to claim 1, it is characterized in that FUFANG YUXINGCAO PIAN is by Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g makes as crude drug, preparation method is comprised of the following step: get Herba Houttuyniae 583g, Radix Scutellariae 150g, Radix Isatidis 150g, Fructus Forsythiae 58g, Flos Lonicerae 58g, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 ℃, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1500, every heavy 0.5g.
5. a kind of FUFANG YUXINGCAO PIAN suppresses the application in rat hepatoma cell RH-35 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in the compound houttuynin piece preparation method
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of FUFANG YUXINGCAO PIAN suppresses the application in rat hepatoma cell RH-35 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in the compound houttuynin piece preparation method
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107496576A (en) * | 2017-09-28 | 2017-12-22 | 南京正宽医药科技有限公司 | A kind of compound houttuynin soft capsule and preparation method and purposes |
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2013
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Non-Patent Citations (2)
| Title |
|---|
| 国家药典委员会: "《中华人民共和国药典一部》", 31 January 2005 * |
| 廖传华等: "《超临界CO2流体萃取技术——工艺开发及其应用》", 31 July 2004 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107496576A (en) * | 2017-09-28 | 2017-12-22 | 南京正宽医药科技有限公司 | A kind of compound houttuynin soft capsule and preparation method and purposes |
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Owner name: HUBEI NORCOTT PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: NANJING ZHENGLIANG PHARMACEUTICAL TECHNOLOGY CO., LTD. Effective date: 20150715 |
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