CN103494933B - Preparation method and application of liver protection tablet - Google Patents
Preparation method and application of liver protection tablet Download PDFInfo
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- CN103494933B CN103494933B CN201310463685.9A CN201310463685A CN103494933B CN 103494933 B CN103494933 B CN 103494933B CN 201310463685 A CN201310463685 A CN 201310463685A CN 103494933 B CN103494933 B CN 103494933B
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Abstract
The invention provides a preparation method of a liver protection tablet. The liver protection tablet is prepared by carrying out supercritical fluid extraction on the following raw herbal materials: 250g of bupleurum falcatum, 250g of capillary artemisia, 250g of isatis root, 300g of schisandra chinensis, 20g of pulvis fellis suis and 128g of mung bean, so that the content is greatly increased, and the dose is reduced. The invention also provides an application of the liver protection tablet in the preparation of a medicine for inhibiting the proliferation of a mouse lymphoma cell YAC-1.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of preparation method and application of liver-protecting tablet.
Background technology
Liver-protecting tablet is recorded in pharmacopeia, and prescription is Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g, above Six-element, and Semen phaseoli radiati powder is broken into fine powder; Radix Bupleuri, Herba Artemisiae Scopariae, Radix Isatidis decoct with water secondary, each 2 hours, filter, merging filtrate, leave standstill 48 hours, get supernatant, concentrating under reduced pressure becomes relative density to be 1.30(80 DEG C) clear paste, mix with Semen phaseoli radiati powder 101g, drying under reduced pressure, be ground into fine powder; Fructus Schisandrae Chinensis powder is broken into coarse powder, with 75% alcohol reflux three times, and 3 hours first times, second time 2 hours, 1 hour third time, merge extractive liquid, leaves standstill 24 hours, get supernatant, reclaim ethanol, being concentrated into relative density is 1.28(80 DEG C), mix with Semen phaseoli radiati powder 27g, drying under reduced pressure, is ground into fine powder; Get Pulvis Fellis Suis, mix with above-mentioned fine powder, granule processed, dry, be pressed into 1000, sugar coating, obtain final product.
In prior art, not yet there is liver-protecting tablet extracting the report adopting supercritical technology in preparation, and adopt the method for beating powder and soak by water, 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.
In prior art, the every sheet 0.5g of liver-protecting tablet, each 5,3 times on the one, adopt the every sheet 0.5g of liver-protecting tablet that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only need 3 at every turn, within 1st, take 3 times, under the condition with more active component, greatly reduce dose.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of preparation method of liver-protecting tablet.
Another object of the present invention is to provide a kind of liver-protecting tablet to suppress the application in mouse lymphoma cell YAC-1 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of liver-protecting tablet, be made up as crude drug of Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g, described method is made up of the following step: get Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g, 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 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
The preparation method of above-mentioned a kind of liver-protecting tablet, 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 liver-protecting tablet, described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of liver-protecting tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation in preparation, liver-protecting tablet is made up as crude drug of Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g, and preparation method is made up of the following step: get Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g 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 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Above-mentioned liver-protecting tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation in preparation, CO described in liver-protecting tablet preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned liver-protecting tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation in preparation, CO described in liver-protecting tablet preparation method
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g join in CO2 supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO
2flow 1m1/g crude drug min, extraction time 150min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 2
Get Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g 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 30MPa, temperature 50 C, CO
2flow 3m1/g crude drug min, extraction time 180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 3
Get Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g 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 20MPa, temperature 40 DEG C, CO
2flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 4: liver-protecting tablet suppresses the experimentation data of YAC-1 cell proliferation
1 experiment material
1.1 experiment cell strains
Mouse lymphoma cell YAC-1 cell, Nanjing Zhengliang Pharmaceutical Technology Co., Ltd.'s laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: liver-protecting tablet of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg liver-protecting tablet, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ l doff pipe subpackages ,-20 DEG C of storages, and simultaneously 0.2 μm of frit dehydrated alcohol is in order to the use of matched group.
1.3 experiment reagent
DMEM(GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang 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); PBS(laboratory autogamy); 1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRA MAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai 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 μm of 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) YAC-1 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, add 5ml complete medium neutralization reaction wherein, proceeded in centrifuge tube after piping and druming cell, the centrifugal 5min of 1000rpm, adjustment concentration of cell suspension 3 × 104/ml.
2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add liver-protecting tablet 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, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.
7) result represents with the suppression ratio of medicine to cell:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 statistical dispositions
Adopt the correlation analysis in Microsoft Excel2003 software and Student t to check, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, 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 pole significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 liver-protecting tablet is to YAC-1 cell inhibitory effect influence research
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Liver-protecting tablet can suppress YAC-1 cell proliferation, and reduce the Growth of Cells number of YAC-1 cell, this effect is dose dependent.
Claims (3)
1. the application of liver-protecting tablet in preparation suppression mouse lymphoma cell YAC-1 cell proliferation, it is characterized in that liver-protecting tablet is made up as crude drug of Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g, preparation method is made up of the following step: get Radix Bupleuri 250g, Herba Artemisiae Scopariae 250g, Radix Isatidis 250g, Fructus Schisandrae Chinensis 300g, Pulvis Fellis Suis 20g, Semen phaseoli radiati 128g 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 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
2. a kind of liver-protecting tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation in preparation according to claim 1, it is characterized in that CO described in liver-protecting tablet preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of liver-protecting tablet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation in preparation according to claim 1, it is characterized in that CO described in liver-protecting tablet preparation method
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102988881A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of pulse unblocking tablet |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102988881A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of pulse unblocking tablet |
Non-Patent Citations (2)
| Title |
|---|
| 国家药典委员会.护肝片.《中华人民共和国药典 2000年版 一部》.化学工业出版社,2000,(第1版),474-475. * |
| 新工艺护肝片的质量标准;全红等;《中国药师》;20101231;第13卷(第5期);660-662 * |
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Owner name: SHANGHAI BAOLONG ANQING PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: NANJING ZHENGLIANG PHARMACEUTICAL TECHNOLOGY CO., LTD. Effective date: 20150610 |
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Inventor after: Ni Jiaquan Inventor after: Jiang Biqing Inventor after: He Lin Inventor before: The inventor has waived the right to be mentioned |
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Effective date of registration: 20150610 Address after: 246001 Yingbin Road 335, Anqing Development Zone, Anhui, China Applicant after: Anqing pharmaceutcal corporation, Ltd of Shanghai Baolong Address before: Lishui Economic Development Zone, Nanjing City, Jiangsu province 211200 zhe Ning Road No. 369 Applicant before: Nanjing Zhengliang Medical Technology Co.,Ltd. |
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