CN104922447A - Health improvement capsule preparation method and application - Google Patents
Health improvement capsule preparation method and application Download PDFInfo
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- CN104922447A CN104922447A CN201510424202.3A CN201510424202A CN104922447A CN 104922447 A CN104922447 A CN 104922447A CN 201510424202 A CN201510424202 A CN 201510424202A CN 104922447 A CN104922447 A CN 104922447A
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Abstract
The invention belongs to the field of Chinese traditional medicine preparation and provides a health improvement capsule preparation method. Health improvement capsules are formed by 75 g of ginseng, 750 g of radix astragali, 500 g of radix ophiopogonis and 2500 g of fructus schizandrae through cationic resin in a supercritical fluid extraction mode, the content is greatly increased, and the use amount is reduced. The invention further provides application of the health improvement capsules in preparing medicine for inhibiting proliferation of immortalized human keratinocyte HaCaT cells.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of preparation method and application of capsule for building up body.
Background technology
Capsule for building up body is recorded in Ministry of Public Health standard, by Radix Ginseng 75g, Fructus Schisandrae Chinensis powder is broken into coarse powder, add 70% alcohol reflux three times, each 2.5 hours, merge extractive liquid, filter, filtrate is concentrated into the thick paste that relative density is 1.3-1.5, medicinal residues and the Radix Astragali, decoct with water three Radix Ophiopogonis, each 2 hours, collecting decoction, filter, filtrate is concentrated into the clear paste that relative density is 1.1-1.15, adding ethanol makes alcohol content reach 60%, stand at low temperature 24 hours, filter, reclaim ethanol, be concentrated into the thick paste that relative density is 1.25-1.35, merge with above-mentioned Fructus Schisandrae Chinensis thick paste, add Radix Ginseng fine powder, mixing, dry, be ground into fine powder, incapsulate, obtain.QI invigorating tonifying YIN, strengthening superficial resistance to stop perspiration, promoting the production of body fluid to quench thirst, for having a delicate constitution caused by deficiency of both QI and YIN, shortness of breath and palpitation, the diseases such as sweating due to debility is thirsty, spiritlessness and weakness.
In prior art, not yet there is capsule for building up body extracting the report adopting cationic resin and supercritical technology in preparation, and adopt the method for pulverizing and 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, former capsule for building up body every 0.25g, 2-3 time on the one, adopt the every sheet 0.25g of capsule for building up body that the present invention is prepared into, but its active component contained increases greatly by each 4.
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 preparation method of capsule for building up body.
Another object of the present invention is to provide a kind of capsule for building up body to suppress the application in people's immortalized keratinocytes HaCaT hyperproliferation agent in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of capsule for building up body, by Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g makes as crude drug, described method is made up of the following step: get Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adding ethanol makes the percent by volume of alcohol content reach 50%, precipitation, leaves standstill, filter, obtain alcohol deposit fluid, alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule, dry, fill, make 1000, every heavy 0.25g.
The percent by volume that described CO2 supercritical extraction entrainer accounts for total extractant is 5%.
The extracting pressure 20MPa of described CO2 supercritical extraction, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
Described capsule for building up body suppresses the application in people's immortalized keratinocytes HaCaT hyperproliferation agent in preparation, described method is made up of the following step: get Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adds ethanol and makes the percent by volume of alcohol content reach 50%, precipitation, leave standstill, filter, obtain alcohol deposit fluid, alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule, dry, fill, make 1000, every heavy 0.25g.
Described capsule for building up body suppresses the application in people's immortalized keratinocytes HaCaT hyperproliferation agent in preparation, and the percent by volume that the entrainer of CO2 supercritical extraction described in the preparation method of capsule for building up body accounts for total extractant is 5%.
Described capsule for building up body suppresses the application in people's immortalized keratinocytes HaCaT hyperproliferation agent in preparation, the extracting pressure 20MPa of CO2 supercritical extraction described in the preparation method of capsule for building up body, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
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 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adding ethanol makes the percent by volume of alcohol content reach 50%, precipitation, leaves standstill, filter, obtain alcohol deposit fluid, alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 50 C, CO2 flow 1m1/g crude drug min, extraction time 180min, obtain supercritical extract, add starch, 70% ethanol granule, dry, fill, make 1000, every heavy 0.25g.
Embodiment 2: get Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adding ethanol makes the percent by volume of alcohol content reach 50%, precipitation, leaves standstill, filter, obtain alcohol deposit fluid, alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa, temperature 30 DEG C, CO2 flow 3m1/g crude drug min, extraction time 150min, obtain supercritical extract, add starch, 70% ethanol granule, dry, fill, make 1000, every heavy 0.25g.
Embodiment 3: get Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adding ethanol makes the percent by volume of alcohol content reach 50%, precipitation, leaves standstill, filter, obtain alcohol deposit fluid, alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, temperature 40 DEG C, CO2 flow 2m1/g crude drug min, extraction time 160min, obtain supercritical extract, add starch, 70% ethanol granule, dry, fill, make 1000, every heavy 0.25g.
Embodiment 4: capsule for building up body suppresses the experimentation data of people's immortalized keratinocytes HaCaT propagation
1 experiment material
1.1 experiment cell strains: people's immortalized keratinocytes HaCaT, Jilin Province just with Pharmaceutical Group Plc laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents: drugs: capsule for building up body of the present invention: prepare by embodiment 3 method.Medicinal liquid liquid storage: take 100mg capsule for building up body content, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ l doff pipe subpackages ,-20 DEG C of storages, and simultaneously 0.2 μm of frit dehydrated alcohol is in order to the use of matched group.
1.3 experiment reagents: DMEM (GIBCO company Cat.No.12100-061 Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3 (Shanghai hundred million chemical reagent company limited Cat.No.11810-033 Lot.No.1088387 of a specified duration); 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 Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS (laboratory autogamy);
1.4 experiment equipments: Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRA MAX 190); 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/N 020579); 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) people's immortalized keratinocytes HaCaT 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, discard culture fluid, PBS fine laundering 3 times, add 3ml 0.25% 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 × 104/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 capsule for building up body solution, continue 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 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 experimental results: statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to people's immortalized keratinocytes HaCaT Proliferation Ability 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 capsule for building up body is to people's immortalized keratinocytes HaCaT Proliferation Ability influence research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
4 experiment conclusion: capsule for building up body can suppress people's immortalized keratinocytes HaCaT to breed, reduce the Growth of Cells number of people's immortalized keratinocytes HaCaT, this effect is dose dependent.
Claims (6)
1. the preparation method of a capsule for building up body, by Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g makes as crude drug, it is characterized in that described method is made up of the following step: get Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adding ethanol makes the percent by volume of alcohol content reach 50%, precipitation, leaves standstill, filter, obtain alcohol deposit fluid; Alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, fill, makes 1000, every heavy 0.25g.
2. the preparation method of capsule for building up body according to claim 1, is characterized in that described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. the preparation method of capsule for building up body according to claim 1, is characterized in that described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
4. capsule for building up body suppresses the application in people's immortalized keratinocytes HaCaT hyperproliferation agent in preparation according to claim 1, it is characterized in that described method is made up of the following step: get Radix Ginseng 75g, Radix Astragali 750g, Radix Ophiopogonis 500g, Fructus Schisandrae Chinensis 2500g, pulverize, add the water boiling and extraction three times of 20,16,14 times amount respectively, each 1 hour, merge three extracting solution, filter, remove slag and get liquid, relative density 1.10 when being concentrated into 65 DEG C, adds ethanol and makes the percent by volume of alcohol content reach 50%, precipitation, leave standstill, filter, obtain alcohol deposit fluid; Alcohol deposit fluid is adsorbed through 10kg cation exchange resin 001 × 7, loading flow velocity is 2ml/min, effluent is for subsequent use, the ammonia of cation exchange resin 10L 0.5mol/L carries out eluting, collect eluent, merge the effluent after cationic exchange resin adsorption and ammonia eluent, reclaim ethanol, join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, fill, makes 1000, every heavy 0.25g.
5. capsule for building up body suppresses the application in people immortalized keratinocytes HaCaT hyperproliferation agent in preparation according to claim 4, CO described in the preparation method that it is characterized in that capsule for building up body
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. capsule for building up body suppresses the application in people immortalized keratinocytes HaCaT hyperproliferation agent in preparation according to claim 4, CO described in the preparation method that it is characterized in that capsule for building up body
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107753733A (en) * | 2017-10-27 | 2018-03-06 | 南京多宝生物科技有限公司 | A kind of capsule for building up body for improving immunity and preparation method thereof |
Citations (2)
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|---|---|---|---|---|
| CN102228513A (en) * | 2011-06-29 | 2011-11-02 | 江苏大学 | Medicinal composition for treating diabetes or diabetic complications and preparation method thereof |
| CN103494902A (en) * | 2013-10-08 | 2014-01-08 | 南京正亮医药科技有限公司 | Preparation method and application of pseudo-ginseng injury tablets |
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- 2015-07-19 CN CN201510424202.3A patent/CN104922447A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102228513A (en) * | 2011-06-29 | 2011-11-02 | 江苏大学 | Medicinal composition for treating diabetes or diabetic complications and preparation method thereof |
| CN103494902A (en) * | 2013-10-08 | 2014-01-08 | 南京正亮医药科技有限公司 | Preparation method and application of pseudo-ginseng injury tablets |
Non-Patent Citations (1)
| Title |
|---|
| 中华人民共和国卫生部药典委员会: "《中华人民共和国卫生部药品标准 中药成方制剂第13册》", 31 December 1997 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107753733A (en) * | 2017-10-27 | 2018-03-06 | 南京多宝生物科技有限公司 | A kind of capsule for building up body for improving immunity and preparation method thereof |
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