CN105998083A - Arenobufagin extraction method and arenobufagin product - Google Patents
Arenobufagin extraction method and arenobufagin product Download PDFInfo
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- CN105998083A CN105998083A CN201610302719.XA CN201610302719A CN105998083A CN 105998083 A CN105998083 A CN 105998083A CN 201610302719 A CN201610302719 A CN 201610302719A CN 105998083 A CN105998083 A CN 105998083A
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- venenum bufonis
- arenobufagin
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- alcohol
- powder
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- 238000000605 extraction Methods 0.000 title claims abstract description 11
- JGDCRWYOMWSTFC-AZGSIFHYSA-N 5-[(3s,5r,8r,9s,10s,11s,13r,14s,17r)-3,11,14-trihydroxy-10,13-dimethyl-12-oxo-2,3,4,5,6,7,8,9,11,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-17-yl]pyran-2-one Chemical compound C=1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)[C@H](O)C3=O)C)C=CC(=O)OC=1 JGDCRWYOMWSTFC-AZGSIFHYSA-N 0.000 title abstract 11
- JGDCRWYOMWSTFC-UHFFFAOYSA-N Arenobufagin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)C(O)C2=O)(O)C2(C)C1C=1C=CC(=O)OC=1 JGDCRWYOMWSTFC-UHFFFAOYSA-N 0.000 title abstract 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 58
- 239000000284 extract Substances 0.000 claims abstract description 28
- 239000003814 drug Substances 0.000 claims abstract description 19
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- 206010018338 Glioma Diseases 0.000 claims abstract description 10
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- 238000009835 boiling Methods 0.000 claims abstract description 6
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- A61K35/56—Materials from animals other than mammals
- A61K35/65—Amphibians, e.g. toads, frogs, salamanders or newts
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- A61K36/185—Magnoliopsida (dicotyledons)
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- 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)
- A61K36/51—Gentianaceae (Gentian family)
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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)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
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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)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- 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
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- 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
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- A—HUMAN NECESSITIES
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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 field of traditional Chinese medicine and discloses an arenobufagin extraction method. The extraction method is characterized by comprising the following steps that a traditional Chinese medicine arenobufagin material is smashed and ground into arenobufagin powder, and the arenobufagin powder is screened through a 15-25-mesh screen; 10-fold ethyl alcohol is used for soaking for 12 h, then, heating and boiling are performed for 2 h and continue to be performed three times, an alcohol extract is subjected to vacuum concentration till no alcohol taste exists, 5% hydrochloric acid is used for dissolution, suction filtration is performed, an acid water solution is extracted repeatedly through petroleum ether, degreasing is performed, strong ammonia water is used for alkalization till pH is 10, the alkaline water solution is extracted two times through trichloromethane, a solvent is recycled, and an arenobufagin crude extract is obtained; the arenobufagin crude extract is dissolved through a chloroform-methanol solution with the volume ratio being 1:1 and passes through an LH-20 gel column, a solvent is recycled, freeze drying is performed, and the arenobufagin extract is obtained. The invention discloses an arenobufagin extract formula, and further discloses application of the arenobufagin extract for preparing medicine for restraining rat gliomas C cells.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to extracting method and the Venenum Bufonis goods of a kind of Venenum Bufonis.
Background technology
Cancer is one of disease maximum to human life's health hazard, has substantial amounts of people to die from cancer every year.Cancer therapy drug
The focus of research and development always study of pharmacy.Having 74% in antitumor drug is natural product or derivatives thereof, such as paclitaxel and
Derivant is exactly the antitumor drug that application effectiveness comparison is good the most clinically.Therefore, from natural product, anticancer compound is found
Or lead compound has great importance.
Summary of the invention
Goal of the invention: in order to solve the problems referred to above, it is an object of the invention to provide extracting method and the Venenum Bufonis goods of a kind of Venenum Bufonis.
Technical scheme: it is an object of the invention to by following scheme realization:
A kind of extracting method of Venenum Bufonis, extraction step is as follows: first by Chinese medicine Venenum Bufonis pulverizing medicinal materials, grind to form Venenum Bufonis powder, described toad
Crisp powder uses after crossing 15～25 mesh sieves；With ethanol 10 times amount soak 12h, then heated and boiled 2h, continuous 3 times, alcohol extract
Be evaporated to without after alcohol taste with 5% dissolving with hydrochloric acid, sucking filtration, aqueous acid through petroleum ether extraction repeatedly, defat, strong aqua ammonia alkali
Changing to pH10, alkaline solution, through chloroform extraction 2 times, recycling design, obtains Venenum Bufonis crude extract, by Venenum Bufonis crude extract body
The chloroform-methanol dissolving for 1:1 of the long-pending ratio, crosses LH-20 gel column, recycling design, lyophilization, obtains Venenum Bufonis extract.
The extracting method of described Venenum Bufonis, is 85-95% for the concentration of alcohol soaked.
A kind of Venenum Bufonis goods, its contained active component is prepared from by following raw materials in part by weight proportioning: Venenum Bufonis extract 1 part,
Cough-relieving grass 50 parts, Herba Swertiae davidi 60 parts, Fructus Malvae 55 parts, Caulis et Folium (Gouaniae Leptostachyae) 40 parts, Herba Origani 60 parts, Venenum Bufonis extract is upper
The method of stating is prepared.
Above-mentioned Venenum Bufonis goods, preparation method step is: take above-mentioned in addition to Venenum Bufonis extract other medical materials merge, boiling twice,
Collecting decoction, when being concentrated into 65 DEG C, relative density is 1.10-1.20, adds ethanol, stirring, makes alcohol content by volume percentage ratio
Calculating reaches 65-85%, stands, and filters, and during filtrate reduced in volume to 65 DEG C, relative density is 1.20-1.30 and reclaims ethanol, will
Concentrated solution is spray-dried, and gained granular powder is broken into dry extract, adds Venenum Bufonis extract and adjuvant, makes required preparation.
Described Venenum Bufonis goods, in preparation method, decocting condition is: amount of water is the 8-12 times amount of medical material weight for the first time, decocts 1-2h,
Amount of water is the 6-10 times amount of medical material weight for the second time, decocts 1-2h.
Described Venenum Bufonis goods, in preparation method, spray drying condition is: inlet temperature is 90-110 DEG C, and leaving air temp is 60-90 DEG C,
Temperature of charge is 60-90 DEG C, and atomizing pressure is 0.2-0.5 MPa, and spray velocity is 1-10ml/s.
The application in preparation suppression rat glioma C6 cell drug of the described Venenum Bufonis goods.
Venenum Bufonis is the secretions of Bufo siccus epidermal gland, and white " milky " liquid is poisonous.Can be used as medicine after drying.
Cough-relieving grass, Classification system Limnophila aromatica (Lam.) Merr.Limnophila gratissima Blume,
With all herbal medicine.The whole year can adopt, and cleans, using fresh herb or dry.Clearing away lung-heat to relieve cough, removing toxic substances and promoting subsidence of swelling.For catching a cold, cough, one hundred days
Cough, venom, ulcerative carbuncle toxic swelling.
Herba Swertiae davidi is the herb of Gentianaceae Swertia plant Herba Swertiae davidi Swertia davidi Franch..Xia Qiu gather, using fresh herb or
Dry.Clearing away lung-heat, parasite killing.For jaundice due to damp-heat, larynx is red and swollen, malignant boil scabies.
Fructus Malvae is the dry mature fruit of Malvaceae plant Semen Abutili Malva verticillata L..When summer, autumn two season fruit maturation
Gather, remove impurity, dry in the shade.Clearing away heat and promoting diuresis, detumescence.For urine retention, edema, thirsty；Urinary tract infection.
Caulis et Folium (Gouaniae Leptostachyae) is Rhamnaceae mouth label platymiscium big bud mouth label Gouania leptostachya DC.var.tonkinensis Pitard.,
It is used as medicine with stem, leaf.The whole year can adopt.Removing pathogenic heat from blood and toxic substance from the body, relaxing muscles and tendons and activating QI and blood in the collateral.For numb limbs and tense tendons；Burn and scald, skin infection are controlled in external.This
Invention is used as medicine with leaf.
Herba Origani is Labiatae Elsholtzia species Herba Origani Elsholtzia ciliata (Thunb.) Hyland. [E.patrini
(Lepech.) Garcke], with all herbal medicine.Summer, autumn heading are tapped when blooming, and remove impurity and dry or using fresh herb.Diaphoresis,
Expelling summer-heat, diuresis.Summer catches a cold, fever without perspiration, heatstroke, and acute gastroenteritis is uncomfortable in chest, halitosis, dysuria.Remaining is medicine
Allusion quotation kind.
Beneficial effect: in the present invention, cough-relieving grass, Herba Swertiae davidi, Fructus Malvae removing toxic substances and promoting subsidence of swelling are monarch drug, Caulis et Folium (Gouaniae Leptostachyae), Herba Origani removing heat from blood solution
Poison is ministerial drug, and Venenum Bufonis is adjuvant, and all medicines are harmonious, and is used for suppressing rat glioma C6 cell.
Detailed description of the invention
Form by the following examples, is described in further detail the foregoing of the present invention again, but should this not understood
Scope for the above-mentioned theme of the present invention is only limitted to Examples below, and all technology realized based on foregoing of the present invention belong to this
The scope of invention.
Embodiment 1: take Venenum Bufonis medical material 1kg and pulverize, grind to form Venenum Bufonis powder, described Venenum Bufonis powder uses after crossing 15 mesh sieves；With 85%
Ethanol 10 times amount soak 12h, then heated and boiled 2h, continuous 3 times, alcohol extract be evaporated to without after alcohol taste with 5% hydrochloric acid
Dissolving, sucking filtration, through petroleum ether extraction repeatedly, defat, strong aqua ammonia alkalizes to pH10 aqueous acid, and alkaline solution is through chloroform
Extract 2 times, recycling design, obtain Venenum Bufonis crude extract, by molten for the chloroform-methanol that Venenum Bufonis crude extract volume ratio is 1:1
Solve, cross LH-20 gel column, recycling design, lyophilization, obtain Venenum Bufonis extract.
Embodiment 2: take Venenum Bufonis medical material 1kg and pulverize, grind to form Venenum Bufonis powder, described Venenum Bufonis powder uses after crossing 25 mesh sieves；With 95%
Ethanol 10 times amount soak 12h, then heated and boiled 2h, continuous 3 times, alcohol extract be evaporated to without after alcohol taste with 5% hydrochloric acid
Dissolving, sucking filtration, through petroleum ether extraction repeatedly, defat, strong aqua ammonia alkalizes to pH10 aqueous acid, and alkaline solution is through chloroform
Extract 2 times, recycling design, obtain Venenum Bufonis crude extract, by molten for the chloroform-methanol that Venenum Bufonis crude extract volume ratio is 1:1
Solve, cross LH-20 gel column, recycling design, lyophilization, obtain Venenum Bufonis extract.
Embodiment 3: take Venenum Bufonis medical material 1kg and pulverize, grind to form Venenum Bufonis powder, described Venenum Bufonis powder uses after crossing 20 mesh sieves；With 90%
Ethanol 10 times amount soak 12h, then heated and boiled 2h, continuous 3 times, alcohol extract be evaporated to without after alcohol taste with 5% hydrochloric acid
Dissolving, sucking filtration, through petroleum ether extraction repeatedly, defat, strong aqua ammonia alkalizes to pH10 aqueous acid, and alkaline solution is through chloroform
Extract 2 times, recycling design, obtain Venenum Bufonis crude extract, by molten for the chloroform-methanol that Venenum Bufonis crude extract volume ratio is 1:1
Solve, cross LH-20 gel column, recycling design, lyophilization, obtain Venenum Bufonis extract.
Embodiment 4: take cough-relieving grass 50g, Herba Swertiae davidi 60g, Fructus Malvae 55g, Caulis et Folium (Gouaniae Leptostachyae) 40g, the mixing of Herba Origani 60g medical material,
Boiling twice, add water 8 times amount into medical material weight for the first time, decocts 1.5h, and add water into medical material weight 6 times for the second time
Amount, decocts 1h, collecting decoction, filters, and filtrate is concentrated into relative density 1.10 (65 DEG C), adds ethanol in proper amount and make alcohol content (body
Fraction) reach 65%, stir, stand 24 hours, filter, filtrate recycling ethanol is also concentrated into relative density 1.10 (65 DEG C)
Extractum, spray-dried (condition be inlet temperature be 100 DEG C, leaving air temp is 80 DEG C, and temperature of charge is 70 DEG C, mist
Changing pressure is 0.2 MPa, and spray velocity is 5ml/s.), it is ground into dry extract, adds the Venenum Bufonis of above-described embodiment 1 preparation
Extract 2g, dextrin 50g, with appropriate 80% ethanol wet, soft material processed, cross 14 mesh sieves and pelletize, and 70 DEG C are dried, and 60 mesh are whole
Grain, obtains granule.
Embodiment 5: take cough-relieving grass 50g, Herba Swertiae davidi 60g, Fructus Malvae 55g, Caulis et Folium (Gouaniae Leptostachyae) 40g, the mixing of Herba Origani 60g medical material,
Boiling twice, the 10 times amount into medical material weight that add water, decoct 2h, second time adds water 8 times amount into medical material weight,
Decocting 1h, collecting decoction, filter, filtrate is concentrated into relative density 1.15 (65 DEG C), adds ethanol in proper amount and make alcohol content (volume
Mark) reach 75%, stir, stand 24 hours, filter, filtrate recycling ethanol is also concentrated into relative density 1.25 (65 DEG C)
Extractum, spray-dried (condition be inlet temperature be 120 DEG C, leaving air temp is 90 DEG C, and temperature of charge is 80 DEG C, mist
Changing pressure is 0.3 MPa, and spray velocity is 7.5/s.), it is ground into dry extract, adds the Venenum Bufonis of above-described embodiment 1 preparation
Extract 2g, starch 50g, mix homogeneously, with appropriate 80% ethanol wet, soft material processed, cross 30 mesh sieves and pelletize, in 70～80 DEG C
Being dried, with 60 mesh sieve granulate, tabletting, sugar coating, subpackage, outer package, censorship is qualified, obtains finished product.
Embodiment 6: take cough-relieving grass 50g, Herba Swertiae davidi 60g, Fructus Malvae 55g, Caulis et Folium (Gouaniae Leptostachyae) 40g, the mixing of Herba Origani 60g medical material,
Boiling twice, add water 10 times amount into medical material weight for the first time, decocts 1.5h, and add water into medical material weight 8 for the second time
Times amount, decocts 1.5h, collecting decoction, filters, and filtrate is concentrated into relative density 1.15 (65 DEG C), adds ethanol in proper amount and make containing alcohol
Amount (volume fraction) reaches 75%, stirs, stands 24 hours, filters, and filtrate recycling ethanol is also concentrated into relative density 1.25
The extractum of (65 DEG C), spray-dried (condition be inlet temperature be 120 DEG C, leaving air temp is 90 DEG C, and temperature of charge is 90 DEG C,
Atomizing pressure is 0.4 MPa, and spray velocity is 7.5/s.), it is ground into dry extract, crosses No. 6 sieves, add above-described embodiment 1
Preparation Venenum Bufonis extract 2g, reinstall No. 1 capsule finished product.
Embodiment 7: the experimentation data of suppression rat glioma C6 cell proliferation
Experiment cell strain: rat glioma C6 cell, Nanjing Medical University's Experimental Animal Center laboratory cell storehouse,
DMEM+10%FBS cellar culture.
Drugs: prepare by embodiment 1 method.
Medicinal liquid liquid storage: weigh the 100mg present invention, is dissolved in 5ml dehydrated alcohol, and 0.2 μm filter filters, 500 μ l doff
Pipe subpackage ,-20 DEG C of storages, 0.2 μm filter filters dehydrated alcohol in case matched group is used simultaneously.
Experiment reagent: DMEM (GIBCO company Cat.No.12100-061Lot.No.758137)；Hyclone (Zhejiang
Sky over the river Hangzhoupro bio tech ltd Lot.No.100419)；NaHCO3 (the long hundred million chemical reagent company limited Cat. in Shanghai
No.11810-033Lot.No.1088387)；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 company limited lot number: 080310182)；MTT (Biosharp lot number: 0793)；PBS (laboratory autogamy)；
Experiment equipment: Lycra inverted microscope (Germany Leica model: DM1L)；Visible-ultraviolet light microwell plate detector is (beautiful
MD company of state model: SPECTRA MAX 190)；CO2 incubator (FORMA model: 3111)；Super-clean bench (Soviet Union
Safe and sound company of clean group manufactures model: SW-CJ-ZFD)；Pure water instrument (Spring company of U.S. model: S/N 020579)；
Accurate pipettor (Gilson Inc of France model: P2)；Electronic balance (Sai Duolisi company limited of Germany model: BT323S)；
Full-automatic high-pressure autoclave (SANYO company of Japan model: MLS-3020)；Table electrothermal air dry oven (upper Nereid
Closely knit test equipment company's 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 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.
Experimental technique: 1) rat glioma C6 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out often
Rule cultivate (10cm culture dish), when cell grows to logarithmic (log) phase, collect cell, discard culture fluid, and PBS rinses 3 times,
Add 3ml 0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, be added thereto to 5ml complete medium
Neutralizing reaction, proceeded in centrifuge tube after piping and druming cell, 1000rpm is centrifuged 5min, adjusts concentration of cell suspension 3 × 104
Individual/ml.2) entering in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator
In (37 DEG C, 5%CO2) cellar culture.3) according to cell growth status, general long to 50%-70%, add the present invention
Solution, continues 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 on shaking table
Vibration 10min, makes crystal fully dissolve.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm.6) simultaneously
Background (being not added with cell, only add culture fluid) is set, control wells (cell, the medicine dissolution medium of same concentrations, culture fluid,
MTT, dimethyl sulfoxide), often group sets 6 multiple holes.7) suppression ratio of cell is represented by result with medicine:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment
It is repeated 3 times.
Statistical disposition: using the correlation analysis in Microsoft Excel 2003 software and Student t inspection, data are with mean
± S.D. represents.
Experimental result: statistical result showed after mtt assay experiment, compares with matched group, when dosage reaches 5mg/ml, to greatly
Ren Mus gland bites chromium glucagonoma PC12 cell inhibitory effect variant (P < 0.05), and dosage this difference when 10mg/ml has significantly
Property (P < 0.01), has pole significant difference (P < 0.001) when dosage reaches 15-20mg/ml.
Table 1 present invention is to rat glioma C6 cell inhibitory effect influence research (X ± SD)
Note: compare with matched group, * P < 0.01；**P<0.001
Experiment conclusion: the present invention can suppress rat glioma C6 cell proliferation, reduces rat glioma C6 thin
The cell growing number of born of the same parents, this effect is dose dependent.
Claims (7)
1. the extracting method of a Venenum Bufonis, it is characterised in that extraction step is as follows: first by Chinese medicine Venenum Bufonis pulverizing medicinal materials,
Grinding to form Venenum Bufonis powder, described Venenum Bufonis powder uses after crossing 15～25 mesh sieves；Soak 12h with ethanol 10 times amount, then heat
Boil 2h, continuous 3 times, alcohol extract be evaporated to without after alcohol taste with 5% dissolving with hydrochloric acid, sucking filtration, aqueous acid is through stone
Oil ether extracts repeatedly, defat, strong aqua ammonia alkalization to pH10, alkaline solution through chloroform extraction 2 times, recycling design,
Obtain Venenum Bufonis crude extract, the chloroform-methanol that Venenum Bufonis crude extract volume ratio is 1:1 dissolved, crosses LH-20 gel column,
Recycling design, lyophilization, obtain Venenum Bufonis extract.
2. the extracting method of Venenum Bufonis as claimed in claim 1, it is characterised in that the concentration of alcohol for soaking is
85-95%.
3. Venenum Bufonis goods, it is characterised in that its contained active component prepared by following raw materials in part by weight proportioning and
Become: Venenum Bufonis extract 1 part, cough-relieving grass 50 parts, Herba Swertiae davidi 60 parts, Fructus Malvae 55 parts, Caulis et Folium (Gouaniae Leptostachyae) 40 parts, soil perfume
Ru 60 parts, Venenum Bufonis extract is that the method for the claims 1 is prepared.
4. as claimed in claim 3 Venenum Bufonis goods, it is characterised in that preparation method step is: take and above-mentioned carry except Venenum Bufonis
Taking other medical materials of beyond the region of objective existence to merge, boiling twice, collecting decoction, when being concentrated into 65 DEG C, relative density is 1.10-1.20,
Add ethanol, stirring, make alcohol content by volume percentage calculation reach 65-85%, stand, filter, filtrate reduced in volume
Being 1.20-1.30 to relative density when 65 DEG C and reclaim ethanol, being spray-dried by concentrated solution, gained granular powder is broken into dry leaching
Cream powder, adds Venenum Bufonis extract and adjuvant, makes required preparation.
5. Venenum Bufonis goods as claimed in claim 4, it is characterised in that in preparation method, decocting condition is: add for the first time
The water yield is the 8-12 times amount of medical material weight, decocts 1-2h, and amount of water is the 6-10 times amount of medical material weight for the second time, decocts
1-2h。
6. Venenum Bufonis goods as claimed in claim 4, it is characterised in that in preparation method, spray drying condition is: air intake
Temperature is 90-110 DEG C, and leaving air temp is 60-90 DEG C, and temperature of charge is 60-90 DEG C, and atomizing pressure is 0.2-0.5 MPa,
Spray velocity is 1-10ml/s.
7. Venenum Bufonis goods application in suppression rat glioma C6 cell drug as claimed in claim 3.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108142933A (en) * | 2018-01-09 | 2018-06-12 | 河南牧农生物科技有限公司 | A kind of extraction process of toad slurry |
| CN109248178A (en) * | 2018-10-19 | 2019-01-22 | 江苏浦金药业有限公司 | A kind of filter method in Venenum Bufonis Injection preparation process |
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| CN105055770A (en) * | 2015-07-27 | 2015-11-18 | 杨宗仙 | Traditional Chinese medicinal composition having anticancer effect |
| CN105477020A (en) * | 2015-06-17 | 2016-04-13 | 赵婷 | Anti-glioma drugs based on Venenum Bufonis extract and preparation method thereof |
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2016
- 2016-05-10 CN CN201610302719.XA patent/CN105998083A/en not_active Withdrawn
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105477020A (en) * | 2015-06-17 | 2016-04-13 | 赵婷 | Anti-glioma drugs based on Venenum Bufonis extract and preparation method thereof |
| CN105055770A (en) * | 2015-07-27 | 2015-11-18 | 杨宗仙 | Traditional Chinese medicinal composition having anticancer effect |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108142933A (en) * | 2018-01-09 | 2018-06-12 | 河南牧农生物科技有限公司 | A kind of extraction process of toad slurry |
| CN109248178A (en) * | 2018-10-19 | 2019-01-22 | 江苏浦金药业有限公司 | A kind of filter method in Venenum Bufonis Injection preparation process |
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