CN104306830A - Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of myeloma cells P3X63Ag8 - Google Patents
Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of myeloma cells P3X63Ag8 Download PDFInfo
- Publication number
- CN104306830A CN104306830A CN201410638879.2A CN201410638879A CN104306830A CN 104306830 A CN104306830 A CN 104306830A CN 201410638879 A CN201410638879 A CN 201410638879A CN 104306830 A CN104306830 A CN 104306830A
- Authority
- CN
- China
- Prior art keywords
- preparation
- extraction
- xuan
- huang
- rhubarb
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- 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
-
- 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
- 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)
- A61K36/28—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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/51—Gentianaceae (Gentian family)
-
- 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)
- A61K36/57—Magnoliaceae (Magnolia family)
-
- 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)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
-
- 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)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/738—Rosa (rose)
-
- 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)
-
- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
-
- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/90—Smilacaceae (Catbrier family), e.g. greenbrier or sarsaparilla
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
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 myeloma cells P3X63Ag8.
Description
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 murine myeloma cell P3X63Ag8 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 murine myeloma cell P3X63Ag8 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 murine myeloma cell P3X63Ag8 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 murine myeloma cell P3X63Ag8 cell proliferation
1. experiment material
1.1 experiment cell strains
Murine myeloma cell P3X63Ag8 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) P3X63Ag8 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 P3X63Ag8 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 P3X63Ag8 cell inhibitory effect influence research (X ± SD)
Group | Drug level (mg/ml) | Suppression ratio (%) |
Matched group | 0 | 0 |
1 | 5 | 13.25±6.56 |
2 | 10 | 25.56±9.87* |
3 | 15 | 35.89±12.79** |
4 | 20 | 43.29±15.67** |
Note: compare with matched group, * P<O.01; * P<0.001.
5. experiment conclusion
Huang-Xuan-Yi-gan sheet can suppress P3X63Ag8 cell proliferation, and reduce the Growth of Cells number of P3X63Ag8 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 murine myeloma cell P3X63Ag8 cell proliferation in preparation.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410638879.2A CN104306830A (en) | 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 myeloma cells P3X63Ag8 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201410638879.2A CN104306830A (en) | 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 myeloma cells P3X63Ag8 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN104306830A true CN104306830A (en) | 2015-01-28 |
Family
ID=52362239
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201410638879.2A Pending CN104306830A (en) | 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 myeloma cells P3X63Ag8 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN104306830A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104306821A (en) * | 2014-11-13 | 2015-01-28 | 山东中大药业有限公司 | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of mastocytoma cell P815 cell |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1496975A (en) * | 2002-09-25 | 2004-05-19 | 上海恒达科技发展股份有限公司 | Emodin derivative with anti-tumour activity |
CN1546103A (en) * | 2003-12-03 | 2004-11-17 | 贵州德祥制药有限责任公司 | Liver benefiting Huangxuan Yigan Powder and its preparing process |
CN102988612A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of tablet for clearing away heat |
-
2014
- 2014-11-13 CN CN201410638879.2A patent/CN104306830A/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1496975A (en) * | 2002-09-25 | 2004-05-19 | 上海恒达科技发展股份有限公司 | Emodin derivative with anti-tumour activity |
CN1546103A (en) * | 2003-12-03 | 2004-11-17 | 贵州德祥制药有限责任公司 | Liver benefiting Huangxuan Yigan Powder and its preparing process |
CN102988612A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of tablet for clearing away heat |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104306821A (en) * | 2014-11-13 | 2015-01-28 | 山东中大药业有限公司 | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of mastocytoma cell P815 cell |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN104382871A (en) | Preparation method of dock root and hemerocallis fulva liver tonifying tablets and application of dock root and hemerocallis fulva liver tonifying tablets to preparation of medicines for inhibiting proliferation of lymphoid neoplasm cell P388D1 | |
CN104306830A (en) | Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of myeloma cells P3X63Ag8 | |
CN104398928A (en) | Preparation method of rumex japonicus houtt-tawny daylily liver-tonifying tablet and application of rumex japonicus houtt-tawny daylily liver-tonifying tablet in preparation of drugs for inhibition of cell proliferation of myeloma cell FO | |
CN104306820A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of lung cancer cell LLC cell | |
CN104306821A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of mastocytoma cell P815 cell | |
CN103735785B (en) | Pure and impure of ferrum broom suppresses the application in lymphoma cell YAC-1 cell proliferation in preparation | |
CN104306828A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of lymphoma cell YAC-1 cell | |
CN104306822A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of liver tumor cell H22 cell | |
CN103705825B (en) | Pure and impure of ferrum broom suppresses the application in teratocarcinoma cell F9 cell proliferation in preparation | |
CN103690863B (en) | The application in check melanin oncocyte B16 cell proliferation prepared by pure and impure of ferrum broom | |
CN104306829A (en) | 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 | |
CN104306824A (en) | Preparing method of rhubarb and day lily liver tonifying tablet and application thereof in preparation of drugs for inhibiting cell proliferation of liver cancer cells Hepa1-6 | |
CN104398810A (en) | Preparation method of nine flavor swertia bimaculata tablet and application of nine flavor swertia bimaculata tablet in preparation of drugs for inhibition of cell proliferation of neoplastic lymphoid cell P388D1 | |
CN104306827A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of myeloma cell SP2/0 cell | |
CN104306826A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of leukemia cell L6565 cell | |
CN104435569A (en) | Preparation method of rumex nepalensis and hemerocallis tablet for benefiting liver and application of rumex nepalensis and hemerocallis tablet to preparation of drug for inhibiting proliferation of melanoma cell B16 | |
CN104434827A (en) | Preparation method of rumex nepalensis and hemerocallis tablet for benefiting liver and application of rumex nepalensis and hemerocallis tablet to preparation of drug for inhibiting proliferation of teratocarcinoma cell P19 | |
CN104435912A (en) | Preparation method of rumex nepalensis and hemerocallis tablet for benefiting liver and application of rumex nepalensis and hemerocallis tablet to preparation of drug for inhibiting proliferation of ascetic tumor cell SAC-IIC3 | |
CN104306825A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of leukemia cell L1210 cell | |
CN104306823A (en) | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of breast tumor cell C127 cell | |
CN104306722A (en) | Preparation method of nine-herb false Chinese swertia herb tablets and application of nine-herb false Chinese swertia herb tablets in preparation of medicament for inhibiting cell proliferation of lymphoma cell EL4 | |
CN104324288A (en) | Preparation method of Huangxuan liver-benefiting tablets and application of Huangxuan liver-benefiting tablets in preparing medicaments for inhibiting cell proliferation of embryonic carcinoma cells F9 | |
CN104383380A (en) | Preparation method of dock root and hemerocallis fulva liver tonifying tablets and application of dock root and hemerocallis fulva liver tonifying tablets to preparation of medicines for inhibiting proliferation of prostate cancer cell RM-1 | |
CN104324289A (en) | Preparation method of Huangxuan liver-benefiting tablets and application of Huangxuan liver-benefiting tablets in preparing medicament for inhibiting cell proliferation of lymphoma cells EL4 | |
CN104398929A (en) | Preparation method of rumex japonicus houtt-tawny daylily liver-tonifying tablet and application of rumex japonicus houtt-tawny daylily liver-tonifying tablet in preparation of drugs for inhibition of cell proliferation of gastric carcinoma cell MFC |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150128 |