CN103536739A - Preparation method and application of Zuogui pill - Google Patents
Preparation method and application of Zuogui pill Download PDFInfo
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- CN103536739A CN103536739A CN201310526829.0A CN201310526829A CN103536739A CN 103536739 A CN103536739 A CN 103536739A CN 201310526829 A CN201310526829 A CN 201310526829A CN 103536739 A CN103536739 A CN 103536739A
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Abstract
The invention provides a preparation method of a Zuogui pill. The Zuogui pill is prepared from the following crude medicines: 200g of prepared rehmannia root, 100g of seed of Chinese dodder, 75g of root of bidentate achyranthes, 100g of glue of tortoise plastron, 100g of deer-horn glue, 100g of yam, 100g of dogwood and 100g of medlar through supercritical extraction. So that content of the Zuogui pill is greatly improved and dosage is reduced. Furthermore, the invention provides an application of the Zuogui pill in preparing a medicine for inhibiting mice lung cancer Lewis cell proliferation.
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 Bolus as a Kidney-Yin-Tonic.
Background technology
Bolus as a Kidney-Yin-Tonic is recorded in health drug standard WS3-B-0053-89 promulgated by the ministries or commissions of the Central Government, prescription is Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, above eight tastes, except Colla cornus cervi, Colla Plastri Testudinis, the Six-elements such as all the other Radix Rehmanniae Preparata are ground into fine powder, sieve and mix.Get Colla cornus cervi, Colla Plastri Testudinis molten, mix with above-mentioned fine powder.Every 100g powder adds refined honey 10g and appropriate water, and general ball is dry, obtains.Can nourishing kidney nourishing YIN.For kidney-YIN being deficient, soreness of waist and knee, night sweat seminal emission, Mental fatigue dry mouth.
In prior art, not yet have Bolus as a Kidney-Yin-Tonic aspect extraction preparation, adopting the report of supercritical technology, and adopt the method for beating powder, 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.
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 Bolus as a Kidney-Yin-Tonic.
Another object of the present invention is to provide a kind of Bolus as a Kidney-Yin-Tonic to suppress the application in mice lung cancer Lewis cell proliferation medicine in preparation.
Technical scheme: the object of the invention is to realize by following scheme:
A kind of preparation method of Bolus as a Kidney-Yin-Tonic, by Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, as crude drug, made, described method is comprised of the following step: get Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, 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 powder, is ground into fine powder, sieves, and mixes.Every 100g Jia Mojia refined honey 100～120g, makes big honeyed pills, obtains.
The preparation method of above-mentioned Bolus as a Kidney-Yin-Tonic, described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned Bolus as a Kidney-Yin-Tonic, described CO
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned Bolus as a Kidney-Yin-Tonic suppresses the application in mice lung cancer Lewis cell proliferation medicine in preparation, Bolus as a Kidney-Yin-Tonic is by Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g makes as crude drug, preparation method is comprised of the following step: get Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, 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 powder, be ground into fine powder, sieve, mix.Every 100g Jia Mojia refined honey 100～120g, makes big honeyed pills, obtains.
Above-mentioned Bolus as a Kidney-Yin-Tonic suppresses the application in mice lung cancer Lewis cell proliferation medicine, CO described in Bolus as a Kidney-Yin-Tonic in preparation
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned Bolus as a Kidney-Yin-Tonic suppresses the application in mice lung cancer Lewis cell proliferation medicine, CO described in Bolus as a Kidney-Yin-Tonic in preparation
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
In prior art, Bolus as a Kidney-Yin-Tonic is oral, 1 ball, 2 times on the one, the heavy 9g of every ball, the every ball 4.5g of Bolus as a Kidney-Yin-Tonic that adopts the present invention to be prepared into, but the medical material amount containing is original 2 times, therefore only need 1 ball at every turn, within 1st, take 2 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 Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, join 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, CO
2flow 1m1/g crude drug min, extraction time 150min, obtains supercritical extract powder, is ground into fine powder, sieves, and mixes.Every 100g Jia Mojia refined honey 100g, makes big honeyed pills, obtains every ball 4.5g.
Embodiment 2
Get Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g 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 powder, is ground into fine powder, sieves, and mixes.Every 100g Jia Mojia refined honey 110g, makes big honeyed pills, obtains every ball 4.5g.
Embodiment 3
Get Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g 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 powder, is ground into fine powder, sieves, and mixes.Every 100g Jia Mojia refined honey 120g, makes big honeyed pills, obtains every ball 4.5g.
Embodiment 4: Bolus as a Kidney-Yin-Tonic suppresses the experimentation data of LEWIS cell proliferation
1 experiment material
1.1 experiment cell strains
Mice lung cancer Lewis cell, Nanjing Medical University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: Bolus as a Kidney-Yin-Tonic of the present invention: press embodiment 3 method preparations.
Medicinal liquid liquid storage: take 100mg Bolus as a Kidney-Yin-Tonic, 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.12100-061 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: SPECTRA MAX 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/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) LEWIS 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 Bolus as a Kidney-Yin-Tonic 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 LEWIS 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 Bolus as a Kidney-Yin-Tonic is on LEWIS cell inhibitory effect impact research
Note: with matched group comparison, * P<0.01; * P<0.001
5 experiment conclusion
Bolus as a Kidney-Yin-Tonic can suppress LEWIS cell proliferation, reduces the Growth of Cells number of LEWIS cell, and this effect is dose dependent.
Claims (6)
1. the preparation method of a Bolus as a Kidney-Yin-Tonic, by Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, as crude drug, made, it is characterized in that described method is comprised of the following step: get Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, 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 powder, is ground into fine powder, sieves, and mixes.Every 100g Jia Mojia refined honey 100～120g, makes big honeyed pills, obtains.
2. a kind of preparation method of Bolus as a Kidney-Yin-Tonic 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 Bolus as a Kidney-Yin-Tonic 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 Bolus as a Kidney-Yin-Tonic suppresses the application in mice lung cancer Lewis cell proliferation medicine in preparation according to claim 1, it is characterized in that Bolus as a Kidney-Yin-Tonic made as crude drug by Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, preparation method is comprised of the following step: get Radix Rehmanniae Preparata 200g, Semen Cuscutae 100g, Radix Achyranthis Bidentatae 75g, Colla Plastri Testudinis 100g, Colla cornus cervi 100g, Rhizoma Dioscoreae 100g, Fructus Corni 100g, Fructus Lycii 100g, 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 powder, is ground into fine powder, sieves, and mixes.Every 100g Jia Mojia refined honey 100～120g, makes big honeyed pills, obtains.
5. a kind of Bolus as a Kidney-Yin-Tonic suppresses the application in mice lung cancer Lewis cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in Bolus as a Kidney-Yin-Tonic
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of Bolus as a Kidney-Yin-Tonic suppresses the application in mice lung cancer Lewis cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in Bolus as a Kidney-Yin-Tonic
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 (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104623197A (en) * | 2015-01-28 | 2015-05-20 | 杭州市萧山区中医院 | Zuogui pill nanoparticle drug delivery system and preparation method thereof |
| CN104887947A (en) * | 2015-06-18 | 2015-09-09 | 南京华宽信息咨询中心 | Preparation method of loins-strengthening kidney-invigorating pill as well as application |
| CN104906059A (en) * | 2015-06-22 | 2015-09-16 | 南京华宽信息咨询中心 | Preparation method and application of anti-inflammatory and antasthmatic tablet |
| CN105641320A (en) * | 2014-11-13 | 2016-06-08 | 上海中医药大学 | Traditional Chinese medicine concentrated pills |
-
2013
- 2013-10-30 CN CN201310526829.0A patent/CN103536739A/en active Pending
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| 吴璇等: "左归丸加黄芪对S180小鼠化疗后卵巢功能损害和抑瘤作用的研究", 《现代中西医结合杂志》 * |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105641320A (en) * | 2014-11-13 | 2016-06-08 | 上海中医药大学 | Traditional Chinese medicine concentrated pills |
| CN104623197A (en) * | 2015-01-28 | 2015-05-20 | 杭州市萧山区中医院 | Zuogui pill nanoparticle drug delivery system and preparation method thereof |
| CN104887947A (en) * | 2015-06-18 | 2015-09-09 | 南京华宽信息咨询中心 | Preparation method of loins-strengthening kidney-invigorating pill as well as application |
| CN104906059A (en) * | 2015-06-22 | 2015-09-16 | 南京华宽信息咨询中心 | Preparation method and application of anti-inflammatory and antasthmatic tablet |
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Application publication date: 20140129 |