CN104887905A - Preparation method for six-ingredient rehmannia pills and application thereof - Google Patents
Preparation method for six-ingredient rehmannia pills and application thereof Download PDFInfo
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- CN104887905A CN104887905A CN201510364117.2A CN201510364117A CN104887905A CN 104887905 A CN104887905 A CN 104887905A CN 201510364117 A CN201510364117 A CN 201510364117A CN 104887905 A CN104887905 A CN 104887905A
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Abstract
The invention belongs to the field of Chinese herbal preparations and particularly provides a preparation method for six-ingredient rehmannia pills. The six-ingredient rehmannia pills are prepared from 160g of prepared rehmannia roots, 80g of processed cornus officinalis, 60g of cortex moutan, 80g of yams, 60g of poria cocos and 60g of rhizoma alismatis by means of supercritical fluid extraction. Therefore, ingredient contents are greatly increased, and dosage of the six-ingredient rehmannia pills is reduced. The invention further provides an application of the six-ingredient rehmannia pills to preparation of medicines for suppressing proliferation of human fibroblasts HF10.
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 LIUWEI DIHUANG WAN.
Background technology
LIUWEI DIHUANG WAN is recorded in Chinese Pharmacopoeia standard, and prescription is Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, and above Six-element, is ground into fine powder, sieves, mixing.Every 100g powder adds refined honey 35 ~ 50g and appropriate water, general ball, dry, makes water-honeyed pill; Or add refined honey 80 ~ 110g and make small honey pill or big honeyed pills, to obtain final product.Enriching yin and nourishing kidney.For damage of kidney-YIN, dizziness and tinnitus, soreness of the waist and knees, osteopyrexia and fever, night sweat is passed out semen, and quenches one's thirst.
In prior art, not yet have LIUWEI DIHUANG WAN extracting the report adopting supercritical technology in preparation, and the methods such as powder are beaten in employing, technique is coarse, complicated, 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 ball 9g of LIUWEI DIHUANG WAN, oral, 1 ball, 2 times on the one, adopt the every ball 9g of LIUWEI DIHUANG WAN that the present invention is prepared into, but the medical material amount contained is more than originally, has more active component when same 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 LIUWEI DIHUANG WAN.
Another object of the present invention is to provide a kind of LIUWEI DIHUANG WAN to suppress the application in human fibroblasts HF10 hyperproliferation agent in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of LIUWEI DIHUANG WAN, by Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g makes as crude drug, described method is made up of the following step: get above-mentioned medical material and 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, be ground into fine powder, sieve, mixing, every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, make 200, every heavy 9g.
The preparation method of described LIUWEI DIHUANG WAN, the percent by volume that described CO2 supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of described LIUWEI DIHUANG WAN, the extracting pressure 20MPa of described CO2 supercritical extraction, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
Described LIUWEI DIHUANG WAN suppresses the application in human fibroblasts HF10 hyperproliferation agent in preparation, LIUWEI DIHUANG WAN is by Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g makes as crude drug, preparation method is: get Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, 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, be ground into fine powder, sieve, mixing, every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, make 200, every heavy 9g.
Described LIUWEI DIHUANG WAN suppresses the application in human fibroblasts HF10 hyperproliferation agent in preparation, and described in the preparation method that it is characterized in that LIUWEI DIHUANG WAN, CO2 supercritical extraction entrainer accounts for the percent by volume of total extractant is 5%.
Described LIUWEI DIHUANG WAN suppresses the application in human fibroblasts HF10 hyperproliferation agent in preparation, the extracting pressure 20MPa of CO2 supercritical extraction described in the preparation method that it is characterized in that LIUWEI DIHUANG WAN, 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: get Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 30MPa, temperature 30 DEG C, CO2 flow 1m1/g crude drug min, extraction time 180min, obtains supercritical extract, be ground into fine powder, sieve, mixing, every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, make 200, every heavy 10g.
Embodiment 2: get Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 15MPa, temperature 50 C, CO2 flow 1m1/g crude drug min, extraction time 180min, obtains supercritical extract, be ground into fine powder, sieve, mixing, every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, make 200, every heavy 10g.
Embodiment 3: get Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, temperature 40 DEG C, CO2 flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, be ground into fine powder, sieve, mixing, every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, make 200, every heavy 10g.
Embodiment 4: LIUWEI DIHUANG WAN suppresses the experimentation data of human fibroblasts HF10 propagation
1 experiment material
1.1 experiment cell strain: human fibroblasts HF10, Nanjing Medical University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents: drugs: LIUWEI DIHUANG WAN of the present invention: prepare by embodiment 3 method.Medicinal liquid liquid storage: take 100mg LIUWEI DIHUANG WAN content, is suspended 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 reagents: DMEM (GIBCO company Cat.No.12100-061 Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3 (Shanghai hundred million chemical reagent company limited Cat.No.11810-033 Lot.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 equipments: Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRA MAX 190); 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/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 (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) HF10 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 3ml 0.25% 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 LIUWEI DIHUANG WAN 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 experimental results: statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to HF10 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 LIUWEI DIHUANG WAN is to human fibroblasts HF10 Proliferation Ability influence research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
4 experiment conclusion: LIUWEI DIHUANG WAN can suppress human fibroblasts HF10 to breed, reduce the Growth of Cells number of human fibroblasts HF10, this effect is dose dependent.
Claims (6)
1. a preparation method for LIUWEI DIHUANG WAN, is made up as crude drug of Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, it is characterized in that described method is made up of the following step: get above-mentioned medical material and 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, is ground into fine powder, sieves, mixing, and every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, makes 200, every heavy 9g.
2. the preparation method of LIUWEI DIHUANG 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. the preparation method of LIUWEI DIHUANG WAN according to claim 1, is characterized in that described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
4. LIUWEI DIHUANG WAN suppresses the application in human fibroblasts HF10 hyperproliferation agent in preparation according to claim 1, it is characterized in that LIUWEI DIHUANG WAN is made up as crude drug of Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, preparation method is: get Radix Rehmanniae Preparata 160g, Fructus Corni (processed) 80g, Cortex Moutan 60g, Rhizoma Dioscoreae 80g, Poria 60g, Rhizoma Alismatis 60g, 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, is ground into fine powder, sieves, mixing, and every 100g powder adds the sweet 120g of white silk and makes big honeyed pills, makes 200, every heavy 9g.
5. LIUWEI DIHUANG WAN suppresses the application in human fibroblasts HF10 hyperproliferation agent in preparation according to claim 4, CO described in the preparation method that it is characterized in that LIUWEI DIHUANG WAN
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. LIUWEI DIHUANG WAN suppresses the application in human fibroblasts HF10 hyperproliferation agent in preparation according to claim 4, CO described in the preparation method that it is characterized in that LIUWEI DIHUANG WAN
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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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105521211A (en) * | 2015-12-28 | 2016-04-27 | 云南中医学院 | Traditional Chinese medicinal ointment rubbing agent for promoting growth of premature low-birth-weight children, and preparation method thereof |
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| CN1634542A (en) * | 2004-11-12 | 2005-07-06 | 康国忠 | Liuwei Dihuang dripping pill preparation and method for preparing the same |
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| CN104000994A (en) * | 2014-06-06 | 2014-08-27 | 余中华 | Preparation method of concentrated six-taste glutinous rehmannia capsule |
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2015
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Patent Citations (4)
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| CN1449806A (en) * | 2003-01-10 | 2003-10-22 | 康国忠 | Technology for producing Liuwei Dihuang Wan |
| CN1634542A (en) * | 2004-11-12 | 2005-07-06 | 康国忠 | Liuwei Dihuang dripping pill preparation and method for preparing the same |
| CN101313972A (en) * | 2007-06-01 | 2008-12-03 | 吴开敏 | Technique for preparing new preparation of six ingredients with rehmannia |
| CN104000994A (en) * | 2014-06-06 | 2014-08-27 | 余中华 | Preparation method of concentrated six-taste glutinous rehmannia capsule |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105521211A (en) * | 2015-12-28 | 2016-04-27 | 云南中医学院 | Traditional Chinese medicinal ointment rubbing agent for promoting growth of premature low-birth-weight children, and preparation method thereof |
| CN105521211B (en) * | 2015-12-28 | 2019-12-13 | 云南中医学院 | Traditional Chinese medicine ointment for promoting growth and development of premature low-weight children and preparation method thereof |
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Application publication date: 20150909 |