CN103520266A - Preparation method and application of pill containing Pogostemon patchouli and pulvis fellis suis - Google Patents
Preparation method and application of pill containing Pogostemon patchouli and pulvis fellis suis Download PDFInfo
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- CN103520266A CN103520266A CN201310472511.9A CN201310472511A CN103520266A CN 103520266 A CN103520266 A CN 103520266A CN 201310472511 A CN201310472511 A CN 201310472511A CN 103520266 A CN103520266 A CN 103520266A
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Abstract
The invention provides a preparation method of a pill containing Pogostemon patchouli and pulvis fellis suis. The pill is prepared from bulk drugs containing 4000 g of Pogostemon patchouli and 315 g of pulvis fellis suis through supercritical extraction, so as to realize increase in content and reduction in the dosage. The invention also provides application of the pill containing Pogostemon patchouli and pulvis fellis suis to preparation of medicines for inhibiting proliferation of human melanoma cell MV3.
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 HUODAN WAN.
Background technology
HUODAN WAN is recorded in pharmacopeia, writes out a prescription as Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, and method for making is ground into fine powder for getting Herba Pogostemonis leaf, sieves; Get Pulvis Fellis Suis alcohol heating reflux, filter, filtrate recycling ethanol, drying under reduced pressure, wears into fine powder, mixes with Herba Pogostemonis leaf fine powder, with water pill, dry, obtains.Heat clearing awayization is turbid, a surname's clearing the nasal passage.For wind and cold heat-transformation, on gallbladder fire, attack the nasal obstruction causing and owe logical, nasosinusitis headache.
In prior art, not yet there is HUODAN WAN aspect extraction preparation, adopting the report of supercritical technology, and adopt the method for beating powder and ethanol extraction, 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 HUODAN WAN.
Another object of the present invention is to provide a kind of HUODAN WAN to suppress the application in human melanoma cell MV3 cell proliferation medicine in preparation.
Technical scheme: the object of the invention is to realize by following scheme:
A kind of preparation method of HUODAN WAN, by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, as crude drug, made, described method is comprised of the following step: get Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 1-3ml/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule processed, dry, make 2500g.
The preparation method of above-mentioned a kind of HUODAN WAN, 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 HUODAN WAN, 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 HUODAN WAN suppresses the application in human melanoma cell MV3 cell proliferation medicine in preparation, HUODAN WAN is made as crude drug by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, preparation method is comprised of the following step: get Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 1-3ml/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, and 70% ethanol granule processed is dry, makes 2500g.
Above-mentioned HUODAN WAN suppresses the application in human melanoma cell MV3 cell proliferation medicine, CO described in HUODAN WAN preparation method in preparation
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned HUODAN WAN suppresses the application in human melanoma cell MV3 cell proliferation medicine, CO described in HUODAN WAN preparation method in preparation
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
The medical material amount containing in the HUODAN WAN that adopts the present invention to be prepared into is original 2 times, and general before each serving consumption being therefore 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 Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 1ml/g crude drug min, extraction time 150min, obtains supercritical extract, add starch, 70% ethanol granule processed, dry, make 2500g.
Embodiment 2
Get Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 3ml/g crude drug min, extraction time 180min, obtains supercritical extract, adds starch, and 70% ethanol granule processed is dry, makes 2500g.
Embodiment 3
Get Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 2ml/g crude drug min, extraction time 160min, obtains supercritical extract, adds starch, and 70% ethanol granule processed is dry, makes 2500g.
Embodiment 4: HUODAN WAN suppresses the experimentation data of MV3 cell proliferation
1 experiment material
1.1 experiment cell strains
Human melanoma cell MV3 cell, Nanjing Zhengkuan Pharmaceutical Technology Co., Ltd.'s laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: HUODAN WAN of the present invention: press embodiment 3 method preparations.
Medicinal liquid liquid storage: take 100mg HUODAN WAN, 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) MV3 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 HUODAN WAN 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 MV3 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 HUODAN WAN is on MV3 cell inhibitory effect impact research
Note: with matched group comparison, * P < 0.01; * P < 0.001
5 experiment conclusion
HUODAN WAN can suppress MV3 cell proliferation, reduces the Growth of Cells number of MV3 cell, and this effect is dose dependent.
Claims (6)
1. a preparation method for HUODAN WAN, is made as crude drug by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, it is characterized in that described method is comprised of the following step: get Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 1-3ml/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, and 70% ethanol granule processed is dry, makes 2500g.
2. a kind of preparation method of HUODAN WAN 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 HUODAN WAN 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 HUODAN WAN suppresses the application in human melanoma cell MV3 cell proliferation medicine in preparation according to claim 1, it is characterized in that HUODAN WAN made as crude drug by Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, preparation method is comprised of the following step: get Herba Pogostemonis leaf 4000g, Pulvis Fellis Suis 315g, 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 1-3ml/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, and 70% ethanol granule processed is dry, makes 2500g.
5. a kind of HUODAN WAN suppresses the application in human melanoma cell MV3 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in HUODAN WAN preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of HUODAN WAN suppresses the application in human melanoma cell MV3 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in HUODAN WAN 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 |
|---|---|---|---|---|
| CN104906056A (en) * | 2015-05-15 | 2015-09-16 | 南方医科大学 | Rhinitis treating dispersible tablet and preparation method thereof |
-
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 2005年版 一部》", 31 December 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 (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104906056A (en) * | 2015-05-15 | 2015-09-16 | 南方医科大学 | Rhinitis treating dispersible tablet and preparation method thereof |
| CN104906056B (en) * | 2015-05-15 | 2018-06-05 | 南方医科大学 | It is a kind of to be used to treat dispersible tablet of rhinitis and preparation method thereof |
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| CN103520266B (en) | 2015-05-13 |
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| ASS | Succession or assignment of patent right |
Owner name: FENG LIAOXING (FOSHAN) PHARMACEUTICAL CO., LTD. OF Free format text: FORMER OWNER: NANJING ZHENGLIANG PHARMACEUTICAL TECHNOLOGY CO., LTD. Effective date: 20150415 |
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Inventor after: Huo Jiayin Inventor after: Lin Shuming Inventor after: Shi Shenhua Inventor after: Liu Guizhong Inventor after: Yang Jianhu Inventor before: The inventor has waived the right to be mentioned |
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Free format text: CORRECT: ADDRESS; FROM: 211200 NANJING, JIANGSU PROVINCE TO: 528000 FOSHAN, GUANGDONG PROVINCE Free format text: CORRECT: INVENTOR; FROM: REQUEST NOT TO RELEASE THE NAME TO: HUO JIAYIN LIN SHUMING SHI SHENHUA LIUGUIZHONG YANG JIANHU |
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Effective date of registration: 20150415 Address after: Chancheng District of Guangdong city in Foshan Province, the 528000 Florida Highway No. 35 of Applicant after: Sinopharm group (Foshan) Pharmaceutical Co., Ltd. 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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Inventor after: Tan Zhen Inventor after: Yang Xionghui Inventor after: Huo Jiayin Inventor after: Lin Shuming Inventor after: Shi Shenhua Inventor after: Liu Guizhong Inventor after: Yang Jianhu Inventor before: Huo Jiayin Inventor before: Lin Shuming Inventor before: Shi Shenhua Inventor before: Liu Guizhong Inventor before: Yang Jianhu |
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Free format text: CORRECT: INVENTOR; FROM: HUO JIAYIN LIN SHUMING SHI SHENHUA LIU GUIZHONG YANG JIANHU TO: TAN ZHEN YANG XIONGHUI HUO JIAYIN LIN SHUMING SHI SHENHUA LIU GUIZHONG YANG JIANHU |