CN102988489B - Preparation method and application of nose comforting tablet - Google Patents
Preparation method and application of nose comforting tablet Download PDFInfo
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- CN102988489B CN102988489B CN201210378257.1A CN201210378257A CN102988489B CN 102988489 B CN102988489 B CN 102988489B CN 201210378257 A CN201210378257 A CN 201210378257A CN 102988489 B CN102988489 B CN 102988489B
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- FXNFHKRTJBSTCS-UHFFFAOYSA-N Baicalein Natural products C=1C(=O)C=2C(O)=C(O)C(O)=CC=2OC=1C1=CC=CC=C1 FXNFHKRTJBSTCS-UHFFFAOYSA-N 0.000 abstract 1
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- 235000017089 Scutellaria baicalensis Nutrition 0.000 abstract 1
- UDFLTIRFTXWNJO-UHFFFAOYSA-N baicalein Chemical compound O1C2=CC(=O)C(O)=C(O)C2=C(O)C=C1C1=CC=CC=C1 UDFLTIRFTXWNJO-UHFFFAOYSA-N 0.000 abstract 1
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- IPQKDIRUZHOIOM-UHFFFAOYSA-N Oroxin A Natural products OC1C(O)C(O)C(CO)OC1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 IPQKDIRUZHOIOM-UHFFFAOYSA-N 0.000 description 10
- IKIIZLYTISPENI-ZFORQUDYSA-N baicalin Chemical compound O1[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 IKIIZLYTISPENI-ZFORQUDYSA-N 0.000 description 10
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- 239000013558 reference substance Substances 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
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- JWHHANVGNNWIRI-UHFFFAOYSA-N methanol phosphoric acid hydrate Chemical compound O.OC.OP(O)(O)=O JWHHANVGNNWIRI-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention provides a preparation method of a nose comforting tablet which is prepared from crude drugs including 100g of scutellaria baicalensis, 150g of Fructus Xanthii, 100g of violet magnolia, 75g of mint, 100g of radix angelicae, 25g of asarum, and 500g of dandelion through supercritical extraction and microwave extraction, so that the content of baicalein is greatly improved. The invention also provides the application of the nose comforting tablet in preparing a medicine for restraining proliferation of human melanoma cell, M14 cell.
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 sharp rhinarium.
Background technology
Profit rhinarium is recorded in Ministry of Public Health standard WS3-B-0746-91, by Radix Scutellariae 100g, Fructus Xanthii 150g, Flos Magnoliae 100g, Herba Menthae 75g, Radix Angelicae Dahuricae 100g, Herba Asari 25g, dandelion 500g, as crude drug, is made, and energy heat-clearing and toxic substances removing, dispels the wind and have one's ideas straightened out.For nasal sinusitis, nasal obstruction watery nasal discharge.
In prior art, not yet favourable rhinarium adopts the report of supercritical and microwave technology extracting aspect preparation, and adopts 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 sharp rhinarium.
Another object of the present invention is to provide a kind of sharp rhinarium to suppress the application in human melanoma cell M14 cell proliferation medicine in preparation.
Technical scheme: the object of the invention is to realize by following scheme:
A kind of preparation method of sharp rhinarium, by Radix Scutellariae 100g, Fructus Xanthii 150g, Flos Magnoliae 100g, Herba Menthae 75g, Radix Angelicae Dahuricae 100g, Herba Asari 25g, dandelion 500g, as crude drug, made, described method is comprised of the following step: get Flos Magnoliae, Herba Menthae, Herba Asari, 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, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 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, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 1000, every heavy 0.5g.
The preparation method of above-mentioned a kind of sharp rhinarium, 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 sharp rhinarium, described microwave extracting power 500W extracts 6 minutes at every turn.
The preparation method of above-mentioned a kind of sharp rhinarium, described CO
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned sharp rhinarium suppresses the application in human melanoma cell M14 cell proliferation medicine in preparation.
In prior art, every 0.5g of sharp rhinarium, each 4,2 times on the one, the every 0.5g of sharp rhinarium that adopts the present invention to be prepared into only needs 2 at every turn, within 1st, takes 2 times, has greatly reduced dose having under the condition of more active component.This conclusion can be by following evidence.
The comparison of baicalin content in sharp rhinarium prepared by test one, distinct methods
1, instrument and reagent the present invention profit rhinarium: press embodiment 3 method preparations, use 1000g crude drug, through extracting, make 1000, every heavy 0.5g.Former sharp rhinarium, by the preparation of ministry standard method, is used 1000g crude drug, through extracting, makes 1000, every heavy 0.5g.Agilent 1200 high performance liquid chromatographs; METTLER AE240 electronic analytical balance; Baicalin reference substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica, be filler; Methanol-water-phosphoric acid (40:60:0.2) is mobile phase; Detection wavelength is 280nm.Number of theoretical plate is pressed baicalin peak and is calculated, and should be not less than 3000.
The preparation of reference substance solution: precision takes at 60 ℃ of drying under reduced pressure baicalin reference substance of 4 hours appropriate, adds methanol and makes every 1ml containing the solution of 18 μ g, obtains.
The preparation of need testing solution: get the present invention's profit rhinarium and former sharp rhinarium, porphyrize, mixes, and gets 1g, accurately weighed, and precision adds 70% ethanol 20ml, close plug, supersound process 10 minutes, centrifugal, get supernatant, obtain.
Algoscopy is accurate reference substance solution and each 20 μ l of need testing solution of drawing respectively, and injection liquid chromatography, measures, and obtains.
3, result
Result shows, in the present invention's profit rhinarium, the content of baicalin is 1.23mg/ sheet; And the content of baicalin is 0.21mg/ sheet in former sharp rhinarium, in the situation that dose reduces, baicalin content improves a lot.
Above-mentioned research shows, the sharp rhinarium that adopts the present invention to prepare, and active constituent content is higher than the standby sharp rhinarium of ministry standard legal system.
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 Scutellariae 100g, Fructus Xanthii 150g, Flos Magnoliae 100g, Herba Menthae 75g, Radix Angelicae Dahuricae 100g, Herba Asari 25g, dandelion 500g, by Flos Magnoliae, Herba Menthae, Herba Asari, 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, 30 ℃ of temperature, CO2 flow 1m1/g crude drug min, extraction time 150min, obtain supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 400W, extracts 2 times, each 4 minutes, combining extraction liquid, concentrated, be added on D101 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, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 1000, every heavy 0.5g.
After testing, in finished product, the content of baicalin is 1.28mg/ sheet.
Embodiment 2
Get Radix Scutellariae 100g, Fructus Xanthii 150g, Flos Magnoliae 100g, Herba Menthae 75g, Radix Angelicae Dahuricae 100g, Herba Asari 25g, dandelion 500g, by Flos Magnoliae, Herba Menthae, Herba Asari, 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, standby; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600W, extracts 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on D101 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, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 1000, every heavy 0.5g.
After testing, in finished product, the content of baicalin is 1.34mg/ sheet.
Embodiment 3
Get Radix Scutellariae 100g, Fructus Xanthii 150g, Flos Magnoliae 100g, Herba Menthae 75g, Radix Angelicae Dahuricae 100g, Herba Asari 25g, dandelion 500g, by Flos Magnoliae, Herba Menthae, Herba Asari, 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, 40 ℃ of temperature, CO
2flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 500W, extracts 2 times, each 6 minutes, combining extraction liquid, concentrated, be added on D101 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, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 1000, every heavy 0.5g.
After testing, in finished product, the content of baicalin is 1.23mg/ sheet.
Embodiment 4: sharp rhinarium suppresses the experimentation data of M14 cell proliferation
1 experiment material
1.1 experiment cell strains
Human melanoma cell M14 cell, Nanjing Zhengkuan Pharmaceutical Technology Co., Ltd.'s laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: the present invention's profit rhinarium: press embodiment 3 method preparations.
Medicinal liquid liquid storage: take 100mg profit rhinarium, be dissolved in 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ l doff pipe subpackages ,-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.12100-061 Lot.No.758137 of DMEM(GIBCO company); Hyclone (Lot.No.100419 of Tian Hang bio tech ltd, Zhejiang); The Cat.No.11810-033 Lot.No.1088387 of NaHCO3(Shanghai Jiu Yi chemical reagent 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: SPECTRAMAX 190); 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/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); 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) M14 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 3ml 0.25% 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 sharp rhinarium 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 Excel 2003 softwares, 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 M14 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.
The sharp rhinarium of table 1 is on M14 cell inhibitory effect impact research
Note: with matched group comparison,
*p<0.01;
*p<0.001
5 experiment conclusion
Profit rhinarium can suppress M14 cell proliferation, reduces the Growth of Cells number of M14 cell, and this effect is dose dependent.
Claims (4)
1. a sharp rhinarium suppresses the application in human melanoma cell M14 cell proliferation medicine in preparation, it is characterized in that sharp rhinarium made as crude drug by Radix Scutellariae 100g, Fructus Xanthii 150g, Flos Magnoliae 100g, Herba Menthae 75g, Radix Angelicae Dahuricae 100g, Herba Asari 25g, dandelion 500g, preparation method is comprised of the following step: get Flos Magnoliae, Herba Menthae, Herba Asari, 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, standby; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 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, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 1000, every heavy 0.5g.
2. a kind of sharp rhinarium suppresses the application in human melanoma cell M14 cell proliferation medicine in preparation according to claim 1, it is characterized in that CO described in preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of sharp rhinarium suppresses the application in human melanoma cell M14 cell proliferation medicine in preparation according to claim 1, it is characterized in that the power of microwave extracting described in preparation method 500W, extracts 6 minutes at every turn.
4. a kind of sharp rhinarium suppresses the application in human melanoma cell M14 cell proliferation medicine in preparation according to claim 1, it is characterized in that CO described in 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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| CN103555784A (en) * | 2013-10-25 | 2014-02-05 | 天津士兰科技有限公司 | Method for simultaneously separating wogonin and baicalein monomers from scutellaria baicalensis |
| CN103656600B (en) * | 2013-12-22 | 2016-03-23 | 李梅 | A kind of preparation method of 12 WENJING WAN and application |
| CN105055589B (en) * | 2015-08-14 | 2018-12-18 | 胡连华 | A kind of nasitis plaster and preparation method thereof |
| CN112245531A (en) * | 2020-11-27 | 2021-01-22 | 佛山市金宝林生物科技有限公司 | Traditional Chinese medicine composition for treating rhinitis and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102397461A (en) * | 2011-11-27 | 2012-04-04 | 苏州派腾生物医药科技有限公司 | Preparation method of tablet for women's health and tranquilness |
| CN102397451A (en) * | 2011-11-26 | 2012-04-04 | 苏州派腾生物医药科技有限公司 | Preparation method of cholagogic tablet |
-
2012
- 2012-10-08 CN CN201210378257.1A patent/CN102988489B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102397451A (en) * | 2011-11-26 | 2012-04-04 | 苏州派腾生物医药科技有限公司 | Preparation method of cholagogic tablet |
| CN102397461A (en) * | 2011-11-27 | 2012-04-04 | 苏州派腾生物医药科技有限公司 | Preparation method of tablet for women's health and tranquilness |
Non-Patent Citations (1)
| Title |
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| 中华人民共和国药典委员会编.利鼻片.《***颁药品标准 中药成方制剂第4册》.1991, * |
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| CN102988489A (en) | 2013-03-27 |
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