CN105267681A - Extraction method and application of plant composition containing ginseng - Google Patents

Abstract

The invention provides an extraction method of a plant composition containing ginseng. The plant composition is prepared from 100 weight parts of ginseng, 60 weight parts of herb of pilosella-leaved drymoglossum, 60 weight parts of Lysimachia decurrens Forst., 80 weight parts of phoenix-tail fern and 50 weight parts of nervilis fordii schlecht as raw materials through a microwave extraction method; with the adoption of the extraction method, the content of the composition is greatly improved, and thus the use amount of the composition is reduced; the invention further provides an application of the plant composition containing ginseng in preparing a medicine for inhibiting mouse B cell hybridoma W6/32 cell proliferation.

Description

A kind of vegetable composition extracting method containing Radix Ginseng and application

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 Ginseng and application.

Background technology

Containing the vegetable composition of Radix Ginseng by Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 parts are made as crude drug, current extracting method decocts with water secondary, each 2-3 hour, collecting decoction, filter, be condensed into thick paste, granulate, dry, sugar coating, to obtain final product.

In prior art, not yet there is the vegetable composition containing Radix Ginseng 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.

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 Ginseng.

Another object of the present invention is to provide a kind of application of vegetable composition in preparation suppression mouse B cell hybridoma W6/32 cell proliferation containing Radix Ginseng.

Technical scheme: the object of the invention is by following scheme realize:

A kind of vegetable composition extracting method containing Radix Ginseng, vegetable composition is by Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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 Radix Ginseng, in preparation method, microwave extracting power is 500W, each extraction 6 minutes.

A kind of vegetable composition containing Radix Ginseng suppresses the application in mouse B cell hybridoma W6/32 cell proliferation in preparation, containing the vegetable composition of Radix Ginseng by Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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 Radix Ginseng suppresses the application in mouse B cell hybridoma W6/32 cell proliferation in preparation, the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes.

In prior art, the every sheet 0.4g of vegetable composition containing Radix Ginseng, each 4,3 times on the one, adopt the every sheet 0.4g of vegetable composition containing Radix Ginseng 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.

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 Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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 Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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 Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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.

Drymoglossum Herba Drymoglossi is that Polypodiaceae Drymoglossum Herba Drymoglossi belongs to small-sized Epiphyte, the antipyretic antiinflammatory of energy, wind dispelling cold expelling, clearing away lung-heat to relieve cough and hemostasis.Classification system is Drymoglossumpiloselloides (L.) C.Presl, heat-clearing and toxic substances removing, hemostasia and detumescence.For jaundice, tuberculous lymphadenitis, parotitis, hemoptysis of pulmonary tuberculosis, metrorrhagia, breast carcinoma, traumatic injury.The present invention is with all herbal medicine.

Scrofula grass is the herb of Primulaceae Lysimachia plant decurrent leaf Herba lysimachiae capillipedis LysimachiadecurrensForst., and autumn gathers, clean, using fresh herb or dry.Promoting blood flow to regulate menstruation, dispersing swelling and dissipating binds.For menoxenia; Lymphoid tuberculosis is controlled in external, injury and bone fracture.

Herba Pteridis Multifidae is a kind of pteridophyta, belongs to Pteridaceae, Pteris.Formal name used at school Pterismultifdapoir.Happiness growth is at concealment, moistening, warm place.Also drought-enduring, in southern winter, the plant arid in warp-February grown in crack of stone, still unlikely completely withered.The main product Yangtze river basin and on the south each province, district, the north Shaanxi, Hebei, Shandong etc. economize also have distribution.Tonifying blood and regulating menstruation, yin nourishing.For menoxenia, deficiency of YIN hectic fever, has a dizzy spell, women's asthenia.The present invention is with all herbal medicine.

Folium Nerviliae fordii is the orchid family Nervilia plant Nervilia fordii Nerviliafordii (Hance) Schlecht., is used as medicine with herb or tuber.Clearing away lung-heat to relieve cough, spleen invigorating removing food stagnancy, sedation-analgesia, heat-clearing and toxic substances removing, dissipating blood stasis for subsidence of swelling.For pulmonary tuberculosis cough spitting of blood, bronchitis, infantile malnutrition, infantile pneumonia, psychosis, treating swelling and pain by traumatic injury, stomatitis, acute laryngitis, sore.The present invention is with all herbal medicine.Radix Ginseng is pharmacopeia kind.

Embodiment 4: the vegetable composition containing Radix Ginseng suppresses the experimentation data of mouse B cell hybridoma W6/32 cell proliferation

1 experiment material

1.1 experiment cell strains

Mouse B cell hybridoma W6/32 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 Ginseng: prepare by embodiment 1 method.

Medicinal liquid liquid storage: take 100mg containing the vegetable composition of Radix Ginseng, 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) mouse B cell hybridoma W6/32 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 Ginseng, 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 mouse B cell hybridoma W6/32 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 Ginseng to mouse B cell hybridoma W6/32 cell inhibitory effect influence research (X ± SD)

Note: compare with matched group, * P<0.01; * P<0.001

5 experiment conclusion

Vegetable composition containing Radix Ginseng can suppress mouse B cell hybridoma W6/32 cell proliferation, and reduce the Growth of Cells number of mouse B cell hybridoma W6/32 cell, this effect is dose dependent.

Claims (4)

1. the vegetable composition extracting method containing Radix Ginseng, it is characterized in that vegetable composition is by Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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 Radix Ginseng 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 mouse B cell hybridoma W6/32 cell proliferation containing the vegetable composition of Radix Ginseng, it is characterized in that the vegetable composition containing Radix Ginseng is by Radix Ginseng 100 parts, Drymoglossum Herba Drymoglossi 60 parts, scrofula grass 60 parts, Herba Pteridis Multifidae 80 parts, Folium Nerviliae fordii 50 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 Radix Ginseng, suppressing the application in mouse B cell hybridoma W6/32 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.