CN105168329A - Extraction method and application of plant composition containing kudzuvine roots - Google Patents
Extraction method and application of plant composition containing kudzuvine roots Download PDFInfo
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- CN105168329A CN105168329A CN201510649153.3A CN201510649153A CN105168329A CN 105168329 A CN105168329 A CN 105168329A CN 201510649153 A CN201510649153 A CN 201510649153A CN 105168329 A CN105168329 A CN 105168329A
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- radix puerariae
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Abstract
The invention provides an extraction method of plant composition containing kudzuvine roots. The plant composition is prepared from raw materials including 100g of kudzuvine roots, 40g of luffa stems, 50g of brucea amarissima and 60g of mysore thorn roots. The plant composition is prepared with a microwave extraction method, the content is greatly improved, and the dosage is reduced. The invention further provides an application of the plant composition containing the kudzuvine roots in preparation of a drug for inhibiting mouse melanoma pulmonary metastasis B16-F10 cell proliferation.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of vegetable composition extracting method containing Radix Puerariae and application.
Background technology
Containing the vegetable composition of Radix Puerariae by Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g makes as crude drug, and current extracting method decocts with water secondary, each 2-3 hour, collecting decoction, filters, is condensed into thick paste, granulate, dry, be pressed into 1000, sugar coating, obtain, energy dredge the meridian passage, relieving cough and resolving phlegm.
In prior art, not yet there is the vegetable composition containing Radix Puerariae extracting the report adopting microwave technology in preparation, and adopt the method for soak by water, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
In prior art, the every sheet 0.5g of vegetable composition containing Radix Puerariae, each 4,3 times on the one, adopt the every sheet 0.5g of vegetable composition containing Radix Puerariae that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only needs 2 at every turn, within 1st, take 3 times, under the condition with more active component, greatly reduce dose.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of vegetable composition extracting method containing Radix Puerariae.
Another object of the present invention is to provide a kind of application of vegetable composition in preparation suppression murine melanoma Lung metastases strain B16-F10 cell proliferation containing Radix Puerariae.
Technical scheme: the object of the invention is by following scheme realize:
A kind of vegetable composition extracting method containing Radix Puerariae, vegetable composition is by Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
Above-mentioned a kind of vegetable composition extracting method containing Radix Puerariae, in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
A kind of vegetable composition containing Radix Puerariae suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation, containing the vegetable composition of Radix Puerariae by Radix Puerariae 100g, Ramulus Luffae 90g, Fructus Bruceae 80g, Radix Caesalpiniae Radicis 150g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
Above-mentioned a kind of vegetable composition containing Radix Puerariae suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation, the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, every sheet 0.4g.
Embodiment 2
Get Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collects 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into capsule.
Embodiment 3
Get Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collects 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into granule.
Ramulus Luffae is dicotyledon medicine cucurbitaceous plant, is used as medicine with mature fruit, fruit network, leaf, rattan, root and seed.Dredge the meridian passage, relieving cough and resolving phlegm.Lumbago, cough, rhinitis, bronchitis.
Fructus Bruceae is the mature seed of quassia Fel Anas domestica subtree.Control dysentery, malaria.For chronic dysentery, chronic dysentery with frequent relapse, malaria.
Radix Caesalpiniae Radicis is root or the root bark of leguminous plant Semen Caesalpiniae.Latin name: Caesalpiniadecapetala (Roth) Alston.Toil, temperature, dispels the wind, cold expelling, dehumidifying.Cure cold cough, general pain, lumbago, laryngalgia, toothache, traumatic injury, lymphogranuloma inguinale inguinal bubo.The present invention is used as medicine with root.All the other are pharmacopeia kind.
Embodiment 4: the vegetable composition containing Radix Puerariae suppresses the experimentation data of murine melanoma Lung metastases strain B16-F10 cell proliferation
1 experiment material
1.1 experiment cell strains
Murine melanoma Lung metastases strain B16-F10 cell, Nanjing Medical University's Experimental Animal Center laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: the present invention is containing the vegetable composition of Radix Puerariae: prepare by embodiment 1 method.
Medicinal liquid liquid storage: take 100mg containing the vegetable composition of Radix Puerariae, be dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff pipe subpackages ,-20 DEG C of storage, 0.2 μm of frit dehydrated alcohol prepares against the use of matched group simultaneously.1.3 experiment reagent
DMEM (GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3 (Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.No.1088387 of a specified duration); Trypsin (AMRESCO company lot number: 2010/04); EDTA (AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt (AMRESCO company lot number: 2010242); StreptomycinSulfate (AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS (laboratory autogamy); 1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μm of filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques
1) murine melanoma Lung metastases strain B16-F10 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% 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, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add the vegetable composition solution containing Radix Puerariae, continue to cultivate 24h.
4) add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.
5) after 4h, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.
7) result represents with the suppression ratio of medicine to cell:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 statistical dispositions
Adopt the correlation analysis in MicrosoftExcel2003 software and Studentt inspection, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to murine melanoma Lung metastases strain B16-F10 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 contains the vegetable composition of Radix Puerariae to murine melanoma Lung metastases strain B16-F10 cell inhibitory effect influence research
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Vegetable composition containing Radix Puerariae can suppress murine melanoma Lung metastases strain B16-F10 cell proliferation, and reduce the Growth of Cells number of murine melanoma Lung metastases strain B16-F10 cell, this effect is dose dependent.
Claims (4)
1. the vegetable composition extracting method containing Radix Puerariae, it is characterized in that vegetable composition is by Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
2. a kind of vegetable composition extracting method containing Radix Puerariae according to claim 1, is characterized in that in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
3. one kind is being suppressed the application in murine melanoma Lung metastases strain B16-F10 cell proliferation containing the vegetable composition of Radix Puerariae, it is characterized in that the vegetable composition containing Radix Puerariae is by Radix Puerariae 100g, Ramulus Luffae 40g, Fructus Bruceae 50g, Radix Caesalpiniae Radicis 60g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
4. a kind of vegetable composition containing Radix Puerariae, suppressing the application in murine melanoma Lung metastases strain B16-F10 cell proliferation, is characterized in that the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes according to claim 3.
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102973667A (en) * | 2012-10-15 | 2013-03-20 | 李正梅 | Preparation method and application of cold-fever tablet |
CN104288642A (en) * | 2014-10-20 | 2015-01-21 | 徐艳 | Traditional Chinese medicine composition for treating bacterial pneumonia as well as preparation method and application thereof |
CN104873872A (en) * | 2015-05-30 | 2015-09-02 | 陈爱华 | Granular formulation for treating perennial allergic rhinitis |
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Publication number | Priority date | Publication date | Assignee | Title |
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CN102973667A (en) * | 2012-10-15 | 2013-03-20 | 李正梅 | Preparation method and application of cold-fever tablet |
CN104288642A (en) * | 2014-10-20 | 2015-01-21 | 徐艳 | Traditional Chinese medicine composition for treating bacterial pneumonia as well as preparation method and application thereof |
CN104873872A (en) * | 2015-05-30 | 2015-09-02 | 陈爱华 | Granular formulation for treating perennial allergic rhinitis |
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Application publication date: 20151223 |