CN103494869B - A kind of preparation method of SHILINTONG PIAN and application - Google Patents
A kind of preparation method of SHILINTONG PIAN and application Download PDFInfo
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- CN103494869B CN103494869B CN201310470470.XA CN201310470470A CN103494869B CN 103494869 B CN103494869 B CN 103494869B CN 201310470470 A CN201310470470 A CN 201310470470A CN 103494869 B CN103494869 B CN 103494869B
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Abstract
The invention provides a kind of preparation method of SHILINTONG PIAN, be made up as crude drug of Herba Desmodii Styracifolii 1000 grams, employing supercritical extraction is prepared from, content is improved a lot, dose reduces, and present invention also offers the application of SHILINTONG PIAN in preparation suppression human lung adenocarcinoma SPC-A-1 cell proliferation.
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 SHILINTONG PIAN.
Background technology
SHILINTONG PIAN is recorded in pharmacopeia, and prescription and technique, for getting Herba Desmodii Styracifolii 1000 grams, decoct with water secondary, each 1.5 hours, collecting decoction, filters, filtrate reduced in volume, add 5 times amount 85% ethanol, fully stir, leave standstill 24 hours, filter, filtrate is condensed into paste, dry, add right amount of auxiliary materials, make granule, dry, tabletted, or sugar coating, to obtain final product.Can clear away the dampness and heat, diuresis and expelling stone.For the puckery pain of pouring drop caused by damp-heat accumulation; Lithangiuria, pyelonephritis has above-mentioned patient.
In prior art, not yet have SHILINTONG PIAN extracting the report adopting supercritical technology in preparation, and adopt the method for soak by water, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, is inconvenient to take, and has had a strong impact on this product and has applied clinically.
In prior art, the every sheet 0.5g of SHILINTONG PIAN, each 4,3 times on the one, adopt the every sheet 0.5g of SHILINTONG PIAN that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only need 2 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 SHILINTONG PIAN.
Another object of the present invention is to provide a kind of SHILINTONG PIAN to suppress the application in human lung adenocarcinoma SPC-A-1 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of SHILINTONG PIAN, be made up as crude drug of Herba Desmodii Styracifolii 1000 grams, described method is made up of the following step: get Herba Desmodii Styracifolii 1000 grams, 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 DEG C, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, add starch, 70% ethanol granule, dry, tabletting, make 500, the heavy 0.5g of every sheet.
The preparation method of above-mentioned a kind of SHILINTONG 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 SHILINTONG PIAN, 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 SHILINTONG PIAN suppresses the application in human lung adenocarcinoma SPC-A-1 cell proliferation in preparation, SHILINTONG PIAN is made up as crude drug of Herba Desmodii Styracifolii 1000 grams, preparation method is made up of the following step: get Herba Desmodii Styracifolii 1000 grams, 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, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule, dry, tabletting, make 500, the heavy 0.5g of every sheet.
Above-mentioned SHILINTONG PIAN suppresses the application in human lung adenocarcinoma SPC-A-1 cell proliferation in preparation, and the percent by volume that the entrainer of CO2 supercritical extraction described in the preparation method of SHILINTONG PIAN accounts for total extractant is 5%.
Above-mentioned SHILINTONG PIAN suppresses the application in human lung adenocarcinoma SPC-A-1 cell proliferation in preparation, the extracting pressure 20MPa of CO2 supercritical extraction described in the preparation method of SHILINTONG PIAN, temperature 40 DEG C, CO2 flow 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
Getting Herba Desmodii Styracifolii 1000 grams joins in CO2 supercritical extraction device, and ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO2 flow 1m1/g crude drug min, extraction time 150min, obtain supercritical extract, add starch, 70% ethanol granule, dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 2
Get Herba Desmodii Styracifolii 1000 grams and 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 Herba Desmodii Styracifolii 1000 grams and 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: SHILINTONG PIAN suppresses the experimentation data of SPC-A-1 cell proliferation
1 experiment material
1.1 experiment cell strains
Human lung adenocarcinoma SPC-A-1 cell, Nanjing Zhengliang Pharmaceutical Technology Co., Ltd.'s laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: SHILINTONG PIAN of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg SHILINTONG PIAN, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff 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); PenicillinGSodiumSalt(AMRESCO company lot number: 2010242); StreptomycinSulfate(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: SPECTRAMAX190); 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) SPC-A-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 SHILINTONG PIAN solution, continue to cultivate 24h.
4) add 20 μ lMTT 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 MicrosoftExcel2003 software and Studentt inspection, 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 SPC-A-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 SHILINTONG PIAN is to SPC-A-1 cell inhibitory effect influence research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
SHILINTONG PIAN can suppress SPC-A-1 cell proliferation, and reduce the Growth of Cells number of SPC-A-1 cell, this effect is dose dependent.
Claims (3)
1. the application of SHILINTONG PIAN in preparation suppression human lung adenocarcinoma SPC-A-1 cell proliferation, it is characterized in that SHILINTONG PIAN is made up as crude drug of Herba Desmodii Styracifolii 1000 grams, preparation method is made up of the following step: get Herba Desmodii Styracifolii 1000 grams, 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, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule, dry, tabletting, make 500, the heavy 0.5g of every sheet.
2. a kind of SHILINTONG PIAN suppresses the application in human lung adenocarcinoma SPC-A-1 cell proliferation in preparation according to claim 1, CO described in the preparation method that it is characterized in that SHILINTONG PIAN
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of SHILINTONG PIAN suppresses the application in human lung adenocarcinoma SPC-A-1 cell proliferation in preparation according to claim 1, CO described in the preparation method that it is characterized in that SHILINTONG PIAN
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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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102988500A (en) * | 2012-10-08 | 2013-03-27 | 卞毓平 | Method for preparing compound Dantong tablets |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102988500A (en) * | 2012-10-08 | 2013-03-27 | 卞毓平 | Method for preparing compound Dantong tablets |
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| 石淋通片的工艺研究;张丽等;《实用中医内科杂志》;20081231;第22卷(第6期);101-102 * |
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