CN105106617A - Extraction method and application of plant composition containing serenoa repens - Google Patents
Extraction method and application of plant composition containing serenoa repens Download PDFInfo
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- CN105106617A CN105106617A CN201510576107.5A CN201510576107A CN105106617A CN 105106617 A CN105106617 A CN 105106617A CN 201510576107 A CN201510576107 A CN 201510576107A CN 105106617 A CN105106617 A CN 105106617A
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Abstract
The invention provides an extraction method of a plant composition containing serenoa repens. The plant composition is prepared from 100 parts of serenoa repens, 50 parts of whorledleaf solomonseal rhizome, 40 parts of pineleaf hedyotis herbs and 70 parts of mallotus apelta by adopting the microwave extraction method, therefore, the content is greatly increased, and the using amount is decreased. The invention further provides an application of the plant composition containing the serenoa repens in preparation of medicine for inhibiting proliferation of xenopus laevis nephrocyte ACTK1.
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 sabal and application.
Background technology
Containing the vegetable composition of sabal by sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts are made as crude drug, and current extracting method decocts with water secondary, each 2-3 hour, collecting decoction, filters, is condensed into thick paste, granulates, drying, sugar coating, to obtain final product.
In prior art, not yet there is the vegetable composition containing sabal 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 0.4g of vegetable composition containing sabal, each 4,3 times on the one, adopt the every sheet 0.4g of vegetable composition containing sabal 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 sabal.
Another object of the present invention is to provide a kind of application of vegetable composition in preparation suppression Africa xenopus kidney ACTK1 cell proliferation containing sabal.
Technical scheme: the object of the invention is by following scheme realize:
A kind of vegetable composition extracting method containing sabal, vegetable composition is by sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts is made 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 sabal, in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
A kind of vegetable composition containing sabal suppresses the application in Africa xenopus kidney ACTK1 cell proliferation in preparation, containing the vegetable composition of sabal by sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts is made 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 sabal suppresses the application in Africa xenopus kidney ACTK1 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 sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts, 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 sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts, 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 capsule.
Embodiment 3
Get sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts, 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 granule.
Sabal, have another name called JUZONG leaf or Serenoa repens, formal name used at school is " sabal ", belongs to Palmae shrub plant, usual growth is in the area of South and North America's hot climate, and the dry fruit of its Serenoa repens Serenoarepens (bartram) small maturation can hyoscine.
Rhizoma Polygonati Verticillati is Liliaceae HUANGJING ZANYU CAPSULE Gansu Rhizoma Polygonati PolygonatumkansuenseMaxim., is used as medicine with root stock.Xia Qiu gathers, and cleans and dries, and place Hui Run, section is dried.Nourishing the liver and kidney, YIN nourishing and the production of body fluid promoting.For neurasthenia, hypertension, headache, throat pain, quenches one's thirst, dry mouth and tougue.
Herba hedyotis pinifoliae is the herb of Maguireothamnus speciosus Folium Pini Auricled Hedyotis Herb.Classification system HedyotispinifoliaWall.exG.Don, reducing swelling and alleviating pain, controls traumatic injury, carbuncle sore, snakebite.
Folium Malloti Apeltae is Euphorbiaceae Mallotus plant Folium Malloti Apeltae Mallotusapelta (Lour.) Muell.Arg., is used as medicine with root and leaf.Root can be adopted the whole year, cleans, and section, dries.The many using fresh herbs of leaf, or the summer, the autumn gather, dry levigation.Root, the leaf of Folium Malloti Apeltae are used as medicine.
Embodiment 4: the vegetable composition containing sabal suppresses the experimentation data of Africa xenopus kidney ACTK1 cell proliferation
1 experiment material
1.1 experiment cell strains
Africa xenopus kidney ACTK1 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 sabal: prepare by embodiment 1 method.
Medicinal liquid liquid storage: take 100mg containing the vegetable composition of sabal, 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) Africa xenopus kidney ACTK1 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 sabal, 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 Africa xenopus kidney ACTK1 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 sabal to Africa xenopus kidney ACTK1 cell inhibitory effect influence research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Vegetable composition containing sabal can suppress Africa xenopus kidney ACTK1 cell proliferation, and reduce the Growth of Cells number of Africa xenopus kidney ACTK1 cell, this effect is dose dependent.
Claims (4)
1. the vegetable composition extracting method containing sabal, it is characterized in that vegetable composition is by sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts is made 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 sabal 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 Africa xenopus kidney ACTK1 cell proliferation containing the vegetable composition of sabal, it is characterized in that the vegetable composition containing sabal is by sabal 100 parts, Rhizoma Polygonati Verticillati 50 parts, Herba hedyotis pinifoliae 40 parts, Folium Malloti Apeltae 70 parts is made 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 sabal, suppressing the application in Africa xenopus kidney ACTK1 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 (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1368234A (en) * | 2001-02-01 | 2002-09-11 | 杨孟君 | Nano medicine 'Ganyou' and its preparing process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1368234A (en) * | 2001-02-01 | 2002-09-11 | 杨孟君 | Nano medicine 'Ganyou' and its preparing process |
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| Title |
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| 刘作梅等: "微波萃取白背叶种子果皮精油的研究", 《湖南农业科学》 * |
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Application publication date: 20151202 |