CN103520294A - Preparation method for Sanhuang tablet - Google Patents
Preparation method for Sanhuang tablet Download PDFInfo
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- CN103520294A CN103520294A CN201310471035.9A CN201310471035A CN103520294A CN 103520294 A CN103520294 A CN 103520294A CN 201310471035 A CN201310471035 A CN 201310471035A CN 103520294 A CN103520294 A CN 103520294A
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Abstract
The invention provides a preparation method for a Sanhuang tablet. The Sanhuang tablet is prepared from the bulk drugs consisting of 300 g of rhubarb root and rhizome, 5 g of berberine hydrochloride and 80 g of baical skullcap root extract through supercritical extraction, so the content of Sanhuang tablet is greatly increased, and the dose of the Sanhuang tablet is reduced. The invention further provides application of the Sanhuang tablet in preparation of drugs used for inhibiting cell proliferation of human large cell carcinoma NCI-H460.
Description
Technical field
The present invention relates to Chinese medicine preparation technical field, be specifically related to a kind of preparation method and application of SANHUANG PIAN.
Background technology
SANHUANG PIAN is recorded in pharmacopeia, writes out a prescription as Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 21g (being equivalent to baicalin 15g), above three tastes, Radix Scutellariae is got by Radix Scutellariae extractum system, decocts with water 1.5 hours for the first time three times, 1 hour for the second time, 40 minutes for the third time, collecting decoction, filter, filtrate adds hydrochloric acid and regulates pH value to 1～2, standing 1 hour, get precipitation, wash with water and make pH value to 5～7, dry, be ground into fine powder, measure content, standby.Get Radix Et Rhizoma Rhei 150g, be ground into fine powder, sieve; Residue Radix Et Rhizoma Rhei powder is broken into coarse powder, add 30% alcohol reflux three times, filter merging filtrate, reclaim ethanol and be evaporated to thick paste shape, add Radix Et Rhizoma Rhei fine powder, berberine hydrochloride fine powder, Radix Scutellariae extractum fine powder and right amount of auxiliary materials, mix granulation, dry, be pressed into 1000, sugar coating, obtains.Energy heat-clearing and toxic substances removing, pathogenic fire purging relieving constipation.For three warmers intenseness of heat, conjunctival congestion and swelling pain, the raw skin ulcer of mouth and nose, laryngopharynx swelling and pain, gingival hemorrhage, restlessness and thirst, yellowish urine constipation; Acute gastroenteritis, dysentery.
In prior art, not yet there is SANHUANG 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 SANHUANG PIAN.
Another object of the present invention is to provide a kind of SANHUANG PIAN to suppress the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation.
Technical scheme: the object of the invention is to realize by following scheme:
A kind of preparation method of SANHUANG PIAN, by Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae 80g makes as crude drug, described method is comprised of the following step: get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g, 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.5g.
The preparation method of above-mentioned a kind of SANHUANG PIAN, described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned a kind of SANHUANG 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 SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation, SANHUANG PIAN is by Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g makes as crude drug, preparation method is comprised of the following step: get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g, 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.5g.
Above-mentioned SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation, and the percent by volume that the entrainer of CO2 supercritical extraction described in SANHUANG PIAN preparation method accounts for total extractant is 5%.
Above-mentioned SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine, the extracting pressure 20MPa of CO2 supercritical extraction described in SANHUANG PIAN preparation method, 40 ℃ of temperature, CO2 flow 2ml/g crude drug min, extraction time 160min in preparation.SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation.
In prior art, every 0.5g of SANHUANG PIAN, each 4,3 times on the one, the every 0.5g of SANHUANG PIAN that adopts the present invention to be prepared into, but the medical material amount containing is original 2 times, therefore only need 2 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 realizing based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g 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, CO2 flow 1m1/g crude drug min, extraction time 150min, obtain supercritical extract, add starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.5g.
Embodiment 2
Get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g joins CO
2in 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, obtains supercritical extract, adds starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.5g.
Embodiment 3
Get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g joins CO
2in 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, obtains supercritical extract, adds starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.5g.
Embodiment 4: SANHUANG PIAN suppresses the experimentation data of NCI-H460 cell proliferation
1 experiment material
1.1 experiment cell strains
The cell lung cancer NCI-H460 of National People's Congress cell, Nanjing Zheng Liang Pharmaceutical Technology Co., Ltd laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: SANHUANG PIAN of the present invention: press embodiment 3 method preparations.
Medicinal liquid liquid storage: take 100mg SANHUANG PIAN, be dissolved in 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ l doff pipe packing ,-20 ℃ of storages, 0.2 μ m filter filters dehydrated alcohol in order to the use of matched group simultaneously.
1.3 experiment reagent
The Cat.No.12100-061Lot.No.758137 of DMEM(GIBCO company); Hyclone (Lot.No.100419 of Tian Hang bio tech ltd, Zhejiang); The NaHCO3(Shanghai Jiu Yi chemical reagent Cat.No.11810-033Lot.No.1088387 of company limited); 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) NCI-H460 cell carries out cellar culture (10cm culture dish) with DMEM+10%FBS in 37 ℃, 5%CO2, 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 3 * 104/ml of concentration of cell suspension.
2) cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 ℃, 5%CO2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add SANHUANG PIAN 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 every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, 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 dispositions
Adopt correlation analysis and Student t check in Microsoft Excel2003 software, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, with matched group comparison, when dosage reaches 5mg/ml, to NCI-H460 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 SANHUANG PIAN is on NCI-H460 cell inhibitory effect impact research-(X ± SD)
Note: with matched group comparison, * P < 0.01; * P < 0.001
5 experiment conclusion
SANHUANG PIAN can suppress NCI-H460 cell proliferation, reduces the Growth of Cells number of NCI-H460 cell, and this effect is dose dependent.
Claims (6)
1. a preparation method for SANHUANG PIAN, is made as crude drug by Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g, it is characterized in that described method is comprised of the following step: get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g, 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 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.5g.
2. a kind of preparation method of SANHUANG PIAN according to claim 1, is characterized in that described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of preparation method of SANHUANG 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 SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation according to claim 1, it is characterized in that SANHUANG PIAN is by Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g makes as crude drug, preparation method is comprised of the following step: get Radix Et Rhizoma Rhei 300g, berberine hydrochloride 5g, Radix Scutellariae extractum 80g, 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule processed, dry, tabletting, make 500, every heavy 0.5g.
5. a kind of SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in SANHUANG PIAN preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of SANHUANG PIAN suppresses the application in the cell lung cancer NCI-H460 of National People's Congress cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in SANHUANG PIAN preparation method
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310471035.9A CN103520294A (en) | 2013-10-11 | 2013-10-11 | Preparation method for Sanhuang tablet |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310471035.9A CN103520294A (en) | 2013-10-11 | 2013-10-11 | Preparation method for Sanhuang tablet |
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| CN103520294A true CN103520294A (en) | 2014-01-22 |
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| CN201310471035.9A Pending CN103520294A (en) | 2013-10-11 | 2013-10-11 | Preparation method for Sanhuang tablet |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105748615A (en) * | 2016-02-28 | 2016-07-13 | 南京正亮医药科技有限公司 | Method for preparing tablet containing rheum officinale, berberine hydrochloride and scutellaria baicalensis and application thereof |
-
2013
- 2013-10-11 CN CN201310471035.9A patent/CN103520294A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 李锦开等主编: "《现代中成药手册》", 31 August 2001, 北京：中国中医药出版社出版 * |
| 柯发敏等: "没食子酸的研究进展", 《泸州医学院学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105748615A (en) * | 2016-02-28 | 2016-07-13 | 南京正亮医药科技有限公司 | Method for preparing tablet containing rheum officinale, berberine hydrochloride and scutellaria baicalensis and application thereof |
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Application publication date: 20140122 |