CN105125761A - Extracting method for plant composition containing valeriana officinalis and application - Google Patents
Extracting method for plant composition containing valeriana officinalis and application Download PDFInfo
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- CN105125761A CN105125761A CN201510643180.XA CN201510643180A CN105125761A CN 105125761 A CN105125761 A CN 105125761A CN 201510643180 A CN201510643180 A CN 201510643180A CN 105125761 A CN105125761 A CN 105125761A
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- radix valerianae
- rhizoma
- vegetable composition
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Abstract
The invention provides an extracting method for a plant composition containing valeriana officinalis. The plant composition is prepared with 100 g of the valeriana officinalis, 60 g of cissus javana, 50 g of coprinopsis atramentaria, 70 g of salix cheilophila roots and 150 g of callicarpa macrophylla leaves as raw medicinal materials. Preparing is carried out with a microwave extraction method, so that the content is greatly increased, and use amount is reduced. The invention further provides an application of the plant composition containing the valeriana officinalis in preparing medicine for inhibiting mice fibroblast L929 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 Rhizoma et radix valerianae and application.
Background technology
Containing the vegetable composition of Rhizoma et radix valerianae by Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 150g make as crude drug, and current extracting method decocts with water secondary, each 2-3 hour, collecting decoction, filters, is condensed into thick paste, granulate, drying, is pressed into 1000, sugar coating, obtain, can heat-clearing and toxic substances removing.
In prior art, not yet there is the vegetable composition containing Rhizoma et radix valerianae 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 Rhizoma et radix valerianae, each 4,3 times on the one, adopt the every sheet 0.4g of vegetable composition containing Rhizoma et radix valerianae 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 Rhizoma et radix valerianae.
Another object of the present invention is to provide a kind of vegetable composition containing Rhizoma et radix valerianae to suppress mice to become application in fiber L929 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of vegetable composition extracting method containing Rhizoma et radix valerianae, vegetable composition is by Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 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 extracting method containing Rhizoma et radix valerianae, in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
A kind of vegetable composition containing Rhizoma et radix valerianae becomes the application in fiber L929 cell proliferation preparation suppression mice, containing the vegetable composition of Rhizoma et radix valerianae by Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 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 Rhizoma et radix valerianae becomes the application in fiber L929 cell proliferation preparation suppression mice, 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 Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 150g, 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 Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 150g, 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 Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 150g, 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.
Herba Cissi Javanae is the herb of vitaceae Herba Cissi Javanae, has dispelling wind to detoxify, detumescence dissipating blood stasis, reunion of bone.Control urticaria, eczema, allergic dermatitis, the fracture injury of tendon and muscle, osteopatia sprain, the effects such as rheumatic numbness.
Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) is the sporophore of Gui San section plant ink-cap (Coprinus atramentartius) or excrement ghost umbrella.Control children's's epilepsy sick, furuncle swells, malignant boil.
Horse Radix Salicis Babylonicae is root and the fibrous root of Salicaceous Plants crow willow.Heat clearing away, pathogenic fire purging, pleasant.Control acute toothache, acute lumbar muscle sprain.
Folium Callicarpae Formosanae is the leaf of Verbenaceae Folium Callicarpae Macrophyllae Callicarpamacrophyllavahl.Astringing to arrest bleeding, heat-clearing and toxic substances removing.
Rhizoma et radix valerianae (Classification system: ValerianaofficinalisL.) Valerianaceae, valeriana is perennial resists cold, bloom, herbaceous plant, Gao Keda 120 centimetres; Root stock tubbiness is head, stem hollow, and stem leaf is avette to width egg shape, and panicle top, umbrella room is raw, and corolla lilac red or white, filigree is flat, the flowering fruit bearing stage 7-9 month.
Embodiment 4: the vegetable composition containing Rhizoma et radix valerianae suppresses mice to become the experimentation data of fiber L929 cell proliferation
1 experiment material
1.1 experiment cell strains
Mice becomes fiber L929 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 Rhizoma et radix valerianae: prepare by embodiment 1 method.
Medicinal liquid liquid storage: take 100mg containing the vegetable composition of Rhizoma et radix valerianae, 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) mice become fiber L929 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 Rhizoma et radix valerianae, 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, fiber L929 cell inhibitory effect variant (P<0.05) is become to mice, 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 becomes fiber L929 cell inhibitory effect influence research containing the vegetable composition of Rhizoma et radix valerianae to mice
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Vegetable composition containing Rhizoma et radix valerianae can suppress mice to become fiber L929 cell proliferation, and reduce the Growth of Cells number that mice becomes fiber L929 cell, this effect is dose dependent.
Claims (4)
1. the vegetable composition extracting method containing Rhizoma et radix valerianae, it is characterized in that vegetable composition is by Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 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.
2. a kind of vegetable composition extracting method containing Rhizoma et radix valerianae according to claim 1, is characterized in that in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
3. the vegetable composition containing Rhizoma et radix valerianae becomes application in fiber L929 cell proliferation suppressing mice, it is characterized in that the vegetable composition containing Rhizoma et radix valerianae is by Rhizoma et radix valerianae 100g, Herba Cissi Javanae 60g, Coprinus atramentarius (Bull.) Fr. (Coprinus sterquilinus Fr.) 50g, horse Radix Salicis Babylonicae 70g, Folium Callicarpae Formosanae 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.
4. a kind of vegetable composition containing Rhizoma et radix valerianae becomes the application in fiber L929 cell proliferation suppression mice according to claim 3, it is characterized in that the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes.
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| CN201510643180.XA CN105125761A (en) | 2015-10-03 | 2015-10-03 | Extracting method for plant composition containing valeriana officinalis and application |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1191137A (en) * | 1998-02-27 | 1998-08-26 | 湖北省老年医学研究所 | Valerian preparation and its preparing process and use |
| CN1337255A (en) * | 2001-09-17 | 2002-02-27 | 上海汇谷生物技术有限公司 | Prepn of common valeriane extractive |
| CN103860802A (en) * | 2014-03-14 | 2014-06-18 | 严白双 | Preparation method and application of Jieguqili tablet |
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2015
- 2015-10-03 CN CN201510643180.XA patent/CN105125761A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1191137A (en) * | 1998-02-27 | 1998-08-26 | 湖北省老年医学研究所 | Valerian preparation and its preparing process and use |
| CN1337255A (en) * | 2001-09-17 | 2002-02-27 | 上海汇谷生物技术有限公司 | Prepn of common valeriane extractive |
| CN103860802A (en) * | 2014-03-14 | 2014-06-18 | 严白双 | Preparation method and application of Jieguqili tablet |
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Application publication date: 20151209 |