CN104306829A - Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of anus sarcoma cells S-180 - Google Patents

Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of anus sarcoma cells S-180 Download PDF

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Publication number
CN104306829A
CN104306829A CN201410638878.8A CN201410638878A CN104306829A CN 104306829 A CN104306829 A CN 104306829A CN 201410638878 A CN201410638878 A CN 201410638878A CN 104306829 A CN104306829 A CN 104306829A
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preparation
extraction
xuan
huang
rhubarb
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权栋栋
王洪涛
刘艳辉
权威宇
胡乃合
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Shandong Zhongda Pharmaceutical Co Ltd
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Shandong Zhongda Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/79—Schisandraceae (Schisandra family)
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/90—Smilacaceae (Catbrier family), e.g. greenbrier or sarsaparilla
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    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin
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    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

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Abstract

The invention belongs to the technical field of traditional Chinese medicines and particularly relates to a preparing method and application of a rhubarb and day lily liver tonifying tablet. According to the preparing method of the rhubarb and day lily liver tonifying tablet, the rhubarb and day lily liver tonifying tablet is prepared from the drug raw materials of 291 g of rhubarb, 103 g of day lily, 92 g of herba senecionis scandentis, 92 g of kiwi fruit, 92 g of smilax ocreafa A. DC, 62 g of wild rose, 62 g of swertia, 62 g of mountain angelica and 62 g of kadsura through supercritical extraction, microwave extraction and macroporous resin absorption, and the emodin content is greatly improved. The invention further provides application of the rhubarb and day lily liver tonifying tablet in the preparation of the drugs for inhibiting the cell proliferation of mouse anus sarcoma cells S-180.

Description

The preparation method of Huang-Xuan-Yi-gan sheet and the application in preparation suppression anus sarcoma cell S-180 cell proliferation thereof
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of preparation method of Huang-Xuan-Yi-gan sheet and suppress the application in mice anus sarcoma cell S-180 cell proliferation in preparation.
Background technology
Liver benefiting Huangxuan Yigan Powder standard No. WS-10465(ZD-0465)-2002, be recorded in national standard for traditional Chinese medicines compilation internal medicine liver and gall fascicle.Be made up as crude drug of Radix Rumicis 291g, Hemerocallis fulva L. 103g, Herba Senecionis Scandentis 92g, Fructus actinidiae chinensis 92g, Rhizoma et Radix smilacis ocreatae 92g, Flos rosae multiflorae 62g, Herba Swertiae bimaculatae 62g, Radix Pimpinellae Candolleanae 62g, Fructus Schisandrae Sphenantherae 62g, there is heat-clearing and toxic substances removing, effect of depressed liver-energy dispersing and function of gallbladder promoting.For the chronic hepatitis B caused by dampness-heat in the liver and gallbladder.
In prior art, not yet there is Huang-Xuan-Yi-gan to be dispersed in and extract preparation aspect employing CO 2the report that supercritical extraction, microwave technology and macroporous adsorption resin technology extract, and the method that powder is got directly beaten by Chinese medicine, 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, liver benefiting Huangxuan Yigan Powder is oral, and be grown up a 9g, 3 times on the one.Liver benefiting Huangxuan Yigan Powder dosage is large.The heavy 0.5g of the every sheet of the Huang-Xuan-Yi-gan sheet adopting the inventive method to be prepared into, only needs 2 at every turn, within 1st, takes 3 times.Dose is greatly reduced under the condition with more active component.This conclusion can be proved by following test.
The comparison of emodin content in Huang-Xuan-Yi-gan sheet prepared by test one, distinct methods
L, instrument and reagent Huang-Xuan-Yi-gan sheet of the present invention: by the preparation of embodiment 1 method, use 918g crude drug, makes 200 through extracting, the heavy 0.5g of every sheet.Former liver benefiting Huangxuan Yigan Powder, according to WS-10465(ZD-0465)-2002 standard method preparations, in contrast.Agilent1200 high performance liquid chromatograph; AE240 electronic analytical balance; Emodin reference substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
Measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VI D).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler; Methanol-0.25% phosphoric acid solution (80: 20) is mobile phase; Determined wavelength is 436nm.Number of theoretical plate calculates should be not less than 3000 by emodin peak.
The preparation of reference substance solution is learnt from else's experience, and to be dried to the emodin reference substance of constant weight appropriate, accurately weighed for phosphorus pentoxide, adds methanol and make the solution of every 1ml containing 0.05mg, to obtain final product.
Huang-Xuan-Yi-gan sheet of the present invention is got in the preparation of product need testing solution of the present invention, and porphyrize, gets 0.4g, accurately weighed, put in apparatus,Soxhlet's, add appropriate amount of ethanol, heating and refluxing extraction is to colourless, and extracting solution is concentrated into about 5ml, lets cool, add dilute hydrochloric acid 5ml, chloroform 30ml, reflux 30 minutes, lets cool, divide and get chloroform solution, the jolting of water liquid chloroform is extracted 2 times (20ml, 15ml), combined chloroform liquid, evaporate to dryness, residue adds methanol to be made dissolving and moves in 10ml measuring bottle, add methanol dilution to scale, shake up, centrifugal, get supernatant, to obtain final product.
The liver benefiting Huangxuan Yigan Powder of this contrast, porphyrize are got in the preparation of reference substance need testing solution, mixing, get 2g, accurately weighed, put in apparatus,Soxhlet's, add appropriate amount of ethanol, heating and refluxing extraction is to colourless, and extracting solution is concentrated into about 5ml, let cool, add dilute hydrochloric acid 5ml, chloroform 30ml, reflux 30 minutes, let cool, divide and get chloroform solution, the jolting of water liquid chloroform is extracted 2 times (20ml, 15ml), combined chloroform liquid, evaporate to dryness, residue adds methanol to be made dissolving and moves in 10ml measuring bottle, adds methanol dilution to scale, shake up, centrifugal, get supernatant, to obtain final product.
Algoscopy is accurate respectively draws each 10 μ l of reference substance solution, product need testing solution of the present invention and reference substance need testing solution, injection liquid chromatography, measures, to obtain final product.
3, result
Result shows, in Huang-Xuan-Yi-gan sheet of the present invention, the content of emodin is 2-4mg/ sheet; And the content of emodin is 0.21mg/ grain in former liver benefiting Huangxuan Yigan Powder, every sheet emodin content is equivalent to the 10-20 of powder every g powder content doubly, and when dose reduces, emodin content improves a lot.
Above-mentioned research shows, adopt Huang-Xuan-Yi-gan sheet prepared by preparation method of the present invention, active constituent content is far away higher than WS-10465(ZD-0465) liver benefiting Huangxuan Yigan Powder prepared of method recorded of-2002 standards.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of preparation method of Huang-Xuan-Yi-gan sheet.
Another object of the present invention is to provide a kind of Huang-Xuan-Yi-gan sheet to suppress the application in mice anus sarcoma cell S-180 cell proliferation in preparation.
The object of the invention is by following scheme realize:
A kind of preparation method of Huang-Xuan-Yi-gan sheet, be made up as crude drug of Radix Rumicis 291g, Hemerocallis fulva L. 103g, Herba Senecionis Scandentis 92g, Fructus actinidiae chinensis 92g, Rhizoma et Radix smilacis ocreatae 92g, Flos rosae multiflorae 62g, Herba Swertiae bimaculatae 62g, Radix Pimpinellae Candolleanae 62g, Fructus Schisandrae Sphenantherae 62g, described preparation method is made up of the following step: get Flos rosae multiflorae, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae, be ground into fine powder, 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 20-30MPa, temperature 30-50 DEG C, CO 2flow l-3ml/g crude drug min, extraction time 130-170min, obtains supercritical extract, for subsequent use, Flos rosae multiflorae after extraction, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae residue and other Six-element medical materials, pulverize, add 60% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extract 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 60% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, by supercritical extract, the mixing of microwave extraction thing adds starch, 60% ethanol granule, dry, tabletting, make 200, the heavy 0.5g of every sheet.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, described CO 2in supercritical extraction, entrainer accounts for the percent by volume of total extractant is 5%.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, described CO 2in supercritical extraction, extracting pressure is 25 MPa.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, described CO 2in supercritical extraction, extraction temperature is 60 DEG C.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, described CO 2cO in supercritical extraction 2flow 2ml/g crude drug min.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, described CO 2in supercritical extraction, extraction time is 150min.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, described microwave extracting power is 700W.
In the preparation method of above-mentioned Huang-Xuan-Yi-gan sheet, each extraction time of described microwave extracting is 8 minutes.
Above-mentioned Huang-Xuan-Yi-gan sheet suppresses the application in mice anus sarcoma cell S-180 cell proliferation in preparation.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further described, so that those skilled in the art more understands the present invention, 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
Fetch earth Radix Et Rhizoma Rhei 291g, Hemerocallis fulva L. 103g, Herba Senecionis Scandentis 92g, Fructus actinidiae chinensis 92g, Rhizoma et Radix smilacis ocreatae 92g, Flos rosae multiflorae 62g, Herba Swertiae bimaculatae 62g, Radix Pimpinellae Candolleanae 62g, Fructus Schisandrae Sphenantherae 62g: get Flos rosae multiflorae, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae, be ground into fine powder, 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 25MPa, temperature 40 DEG C, CO 2flow 2ml/g crude drug min, extraction time 150min, obtains supercritical extract, for subsequent use, Flos rosae multiflorae after extraction, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae residue and other Six-element medical materials, pulverize, add 60% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 700W, extract 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 60% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, by supercritical extract, the mixing of microwave extraction thing adds starch, 60% ethanol granule, dry, tabletting, make 200, the heavy 0.5g of every sheet.
After testing, in finished product, the content of emodin is 4.02mg/ sheet.
Embodiment 2
Fetch earth Radix Et Rhizoma Rhei 291g, Hemerocallis fulva L. 103g, Herba Senecionis Scandentis 92g, Fructus actinidiae chinensis 92g, Rhizoma et Radix smilacis ocreatae 92g, Flos rosae multiflorae 62g, Herba Swertiae bimaculatae 62g, Radix Pimpinellae Candolleanae 62g, Fructus Schisandrae Sphenantherae 62g: get Flos rosae multiflorae, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae, be ground into fine powder, join CO 2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 30MPa, temperature 50 C, CO 2flow 3ml/g crude drug min, extraction time 130min, obtains supercritical extract, for subsequent use, Flos rosae multiflorae after extraction, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae residue and other Six-element medical materials, pulverize, add 60% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600W, extract 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 60% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, by supercritical extract, the mixing of microwave extraction thing adds starch, 60% ethanol granule, dry, tabletting, make 200, the heavy 0.5g of every sheet.
After testing, in finished product, the content of emodin is 2.03mg/ sheet.
Embodiment 3
Fetch earth Radix Et Rhizoma Rhei 291g, Hemerocallis fulva L. 103g, Herba Senecionis Scandentis 92g, Fructus actinidiae chinensis 92g, Rhizoma et Radix smilacis ocreatae 92g, Flos rosae multiflorae 62g, Herba Swertiae bimaculatae 62g, Radix Pimpinellae Candolleanae 62g, Fructus Schisandrae Sphenantherae 62g: get Flos rosae multiflorae, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae, be ground into fine powder, 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 20MPa, temperature 30 DEG C, CO 2flow 1ml/g crude drug min, extraction time 170min, obtains supercritical extract, for subsequent use, Flos rosae multiflorae after extraction, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae residue and other Six-element medical materials, pulverize, add 60% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 800W, extract 2 times, each 10 minutes, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 60% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, by supercritical extract, the mixing of microwave extraction thing adds starch, 60% ethanol granule, dry, tabletting, make 200, the heavy 0.5g of every sheet.
After testing, in finished product, the content of emodin is 3.17mg/ sheet.
Embodiment 4: Huang-Xuan-Yi-gan sheet suppresses the experimentation data of mice anus sarcoma cell S-180 cell proliferation
1. experiment material
1.1 experiment cell strains
Mice anus sarcoma cell S-180 cell, Shandong University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: Huang-Xuan-Yi-gan sheet of the present invention: prepare by embodiment 1 method.
Medicinal liquid liquid storage: take 100mg Huang-Xuan-Yi-gan sheet, 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.12100-061 Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHC0 3(Shanghai hundred million chemical reagent company limited Cat.No.11810-033 Lot.No. 1088387 of a specified duration); Trypsin(AMRESCO company); EDTA(AMRESCO company); Penicillin G Sodium Salt(AMRESCO company 1); Streptomycin Sulfate (AMRESCO); Dehydrated alcohol (Zibo Ya Dulan Trade Co., Ltd.); MTT (Biosharp lot number: 0793): PBS(laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DMIL); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX 190); C0 2incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Sprlng 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); 1cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2. experimental technique
1) S-180 cell DMEM+10%FBS is in 37 DEG C, 5%C0 2carry 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 × 10 4individual/ml.
2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 DEG C, 5%C0 2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add Huang-Xuan-Yi-gan sheet 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, as entered 200 μ l dimethyl sulfoxide, is put 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 disposition
Adopt the correlation analysis in Microsoft Excel 2007 software and Student t to check, data represent with mean ± S.D..
4. experimental result
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to S-180 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 Huang-Xuan-Yi-gan sheet is to S-180 cell inhibitory effect influence research (X ± SD)
Group Drug level (mg/ml) Suppression ratio (%)
Matched group 0 0
1 5 10.89±5.55
2 10 20.06±8.69*
3 15 38.24±11.27**
4 20 46.95±15.02**
Note: compare with matched group, * P<O.01; * P<0.001.
5. experiment conclusion
Huang-Xuan-Yi-gan sheet can suppress S-180 cell proliferation, and reduce the Growth of Cells number of S-180 cell, this effect is dose dependent.

Claims (9)

1. the preparation method of a Huang-Xuan-Yi-gan sheet, be made up as crude drug of Radix Rumicis 291g, Hemerocallis fulva L. 103g, Herba Senecionis Scandentis 92g, Fructus actinidiae chinensis 92g, Rhizoma et Radix smilacis ocreatae 92g, Flos rosae multiflorae 62g, Herba Swertiae bimaculatae 62g, Radix Pimpinellae Candolleanae 62g, Fructus Schisandrae Sphenantherae 62g, it is characterized in that, described preparation method is made up of the following step: get Flos rosae multiflorae, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae, be ground into fine powder, 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 20-30MPa, temperature 30-50 DEG C, CO 2flow l-3ml/g crude drug min, extraction time 130-170min, obtains supercritical extract, for subsequent use, Flos rosae multiflorae after extraction, Radix Pimpinellae Candolleanae, Fructus Schisandrae Sphenantherae residue and other Six-element medical materials, pulverize, add 60% ethanol of 5L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extract 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 60% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, by supercritical extract, the mixing of microwave extraction thing adds starch, 60% ethanol granule, dry, tabletting, make 200, the heavy 0.5g of every sheet.
2. the preparation method of the Huang-Xuan-Yi-gan sheet according to claim l, is characterized in that, described CO 2in supercritical extraction, entrainer accounts for the percent by volume of total extractant is 5%.
3. the preparation method of the Huang-Xuan-Yi-gan sheet according to claim l, is characterized in that, described CO 2in supercritical extraction, extracting pressure is 25MPa.
4. the preparation method of the Huang-Xuan-Yi-gan sheet according to claim l, is characterized in that, described CO 2in supercritical extraction, extraction temperature is 60 DEG C.
5. the preparation method of the Huang-Xuan-Yi-gan sheet according to claim l, is characterized in that, described CO 2cO in supercritical extraction 2flow 2ml/g crude drug min.
6. according to the preparation method of the Huang-Xuan-Yi-gan sheet described in claim l, it is characterized in that, described CO 2in supercritical extraction, extraction time is 150min.
7. the preparation method of the Huang-Xuan-Yi-gan sheet according to claim l, is characterized in that, described microwave extracting power is 700W.
8. the preparation method of the Huang-Xuan-Yi-gan sheet according to claim l, is characterized in that, each extraction time of described microwave extracting is 8 minutes.
9. the Huang-Xuan-Yi-gan sheet described in claim l suppresses the application in mice anus sarcoma cell S-180 cell proliferation in preparation.
CN201410638878.8A 2014-11-13 2014-11-13 Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of anus sarcoma cells S-180 Pending CN104306829A (en)

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