CN103566023A - Preparation method and application of toothache anti-inflammation tablets - Google Patents
Preparation method and application of toothache anti-inflammation tablets Download PDFInfo
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Abstract
The invention belongs to the technical field of traditional Chinese medicaments, and in particular relates to a preparation method and application of toothache anti-inflammation tablets. The toothache anti-inflammation tablets are prepared from the following raw materials: 200g of gypsum, 80g of herba schizonepetae, 80g of divaricate saposhnikovia root, 100g of pericarpium citri reticulatae viride, 100g of paeonia suffruticosa, 150g of rehmannia, 100g of indigo naturalis, 60g of asarum, 50g of angelica root and 60g of liquorice by adopting supercritical extraction and microwave extraction, so that the content of hesperidin is greatly increased. The invention also provides application of the toothache anti-inflammation tablets in preparation of medicaments for inhibiting L1210 cell multiplication of leukemia cells of mice.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of preparation method and application of toothache Pianomide.
background technology
Toothache antiphlogistic granules is recorded in one the 807th page of the tenth the 85th page of Traditional Chinese medicine historical preparation and Chinese Pharmacopoeia version in 2010, by Gypsum Fibrosum 200g, Herba Schizonepetae 80g, Radix Saposhnikoviae 80g, Pericarpium Citri Reticulatae Viride 100g, Cortex Moutan 100g Radix Rehmanniae 150g, Indigo Naturalis 100g, Herba Asari 60g, Radix Angelicae Dahuricae 50g, Radix Glycyrrhizae 60g, as crude drug, made, there is wind and heat dispersing, the effect of cooling blood and relieving pain.For headache fever of the body, xerostomia halitosis, constipation dryness accumulated in the stomach and intestine, the gingival swelling and pain due to attacking on retention of heat in the spleen and stomach, wind heat; Acute tooth root periapical periodontitis, pericoronitis of the wisdom tooth, acute gingiva (week) inflammation, acute pulpitis are shown in above-mentioned patient.
In prior art, not yet there is toothache antiphlogistic granules aspect extraction preparation, adopting the report of supercritical and microwave technology, and volatile oil extracts, dipping diafiltration, the method that decocting boils, 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, each 1 bag of toothache antiphlogistic granules, every bag of 20g(has sugar) or 10g(sugar-free), 3 times on the one, double first.Toothache antiphlogistic granules dosage is large, and more bitter under the state of not sugaring, and patient is not easy under clothes, and compliance is poor.The every 0.8g of toothache Pianomide that adopts the inventive method to be prepared into only needs 2 at every turn, within 1st, takes 3 times, doubles first.Greatly reduced dose having under the condition of more active component.This conclusion can be by following evidence.And be prepared into tablet, patient swallows with water, dosage is little and without bitterness, patient is easy to accept.
The comparison of content of hesperidin in toothache Pianomide prepared by test one, distinct methods
L, instrument and reagent toothache Pianomide of the present invention: press embodiment 1 method preparation, use 980g crude drug, make 100 through extracting, every heavy 0.8g.Former toothache antiphlogistic granules, by 2010 editions standards of pharmacopoeia method preparations, is used 980g crude drug, through extracting, makes 50 bags, every bag heavy 10g.Agilent1200 high performance liquid chromatograph; METTLER AE240 electronic analytical balance; Hesperidin reference substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
According to high performance liquid chromatography (appendix V D), measure.
Chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filler; The methanol-water (35:65) of take is mobile phase; Detection wavelength is 283nm.Number of theoretical plate calculates and should be not less than 1000 by Hesperidin peak.
It is appropriate that Hesperidin reference substance is got in the preparation of reference substance solution, accurately weighed, adds methanol and make every 1ml containing the solution of 80 g, obtains.
Toothache Pianomide of the present invention is got in the preparation of product need testing solution of the present invention, and porphyrize, gets 0.16g, accurately weighed, to put in tool plug conical flask, precision adds the close plug of methanol 25ml., weighed weight, supersound process (power 250w. frequency 33kHz) 1 hour, lets cool, weighed weight again, the weight of supplying less loss with methanol, shakes up, and filters, get subsequent filtrate, obtain.
The toothache antiphlogistic granules of this contrast is got in the preparation of reference product need testing solution, and porphyrize, gets 1g, accurately weighed, to put in tool plug conical flask, precision adds the close plug of methanol 25ml., weighed weight, supersound process (power 250w. frequency 33kHz) 1 hour, lets cool, weighed weight again, the weight of supplying less loss with methanol, shakes up, and filters, get subsequent filtrate, obtain.
Algoscopy is accurate reference substance solution and each 20 l. injection liquid chromatographies of need testing solution drawn respectively, measure, and obtain.
3, result
Result shows, in toothache Pianomide of the present invention, the content of Hesperidin is 75-90mg/ sheet; And the content of Hesperidin is 16.2mg/ bag in former toothache antiphlogistic granules, in the situation that dose reduces, content of hesperidin improves a lot.
Above-mentioned research shows, the toothache Pianomide that adopts preparation method of the present invention to prepare, the toothache antiphlogistic granules that active constituent content is prepared higher than the method for recording in 2010 editions standards of pharmacopoeia far away.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of preparation method of toothache Pianomide.
Another object of the present invention is to provide a kind of toothache Pianomide to suppress the application in mouse leukemia cell L1210 cell proliferation medicine in preparation.
The object of the invention is to realize by following scheme:
A kind of preparation method of toothache Pianomide, by Gypsum Fibrosum 200g, Herba Schizonepetae 80g, Radix Saposhnikoviae 80g, Pericarpium Citri Reticulatae Viride 100g, Cortex Moutan 100g, Radix Rehmanniae 150g, Indigo Naturalis 100g, Herba Asari 60g, Radix Angelicae Dahuricae 50g, Radix Glycyrrhizae 60g, as crude drug, made, described preparation method is comprised of the following step: get Herba Schizonepetae, Herba Asari, the Radix Angelicae Dahuricae, Indigo Naturalis, 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 ℃, CO
2flow l-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 50% ethanol of 1.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extracts 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 100, every heavy 0.8g.
In the preparation method of above-mentioned toothache Pianomide, described CO
2in supercritical extraction, to account for the percent by volume of total extractant be 5% to entrainer.
In the preparation method of above-mentioned toothache Pianomide, described CO
2in supercritical extraction, extracting pressure is 25 MPa.
In the preparation method of above-mentioned toothache Pianomide, described CO
2in supercritical extraction, extraction temperature is 60 ℃.
In the preparation method of above-mentioned toothache Pianomide, described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
In the preparation method of above-mentioned toothache Pianomide, described CO
2in supercritical extraction, extraction time is 200min.
In the preparation method of above-mentioned toothache Pianomide, described microwave extracting power is 700W.
In the preparation method of above-mentioned toothache Pianomide, the each extraction time of described microwave extracting is 8 minutes.
Above-mentioned toothache Pianomide suppresses the application in mouse leukemia cell L1210 cell proliferation medicine in preparation.
The specific embodiment
Below in conjunction with specific embodiment, the present invention is further described, so that those skilled in the art more understands the present invention, but this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following example, all technology realizing based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Gypsum Fibrosum 200g, Herba Schizonepetae 80g, Radix Saposhnikoviae 80g, Pericarpium Citri Reticulatae Viride 100g, Cortex Moutan 100g, Radix Rehmanniae 150g, Indigo Naturalis 100g, Herba Asari 60g, Radix Angelicae Dahuricae 50g, Radix Glycyrrhizae 60g, Herba Schizonepetae, Herba Asari, the Radix Angelicae Dahuricae, Indigo Naturalis are joined to CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 25MPa, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 200min, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 50% ethanol of 1.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 700W, extracts 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 100, every heavy 0.8g.
After testing, in finished product, the content of Hesperidin is 88.5mg/ sheet.
Embodiment 2
Get Gypsum Fibrosum 200g, Herba Schizonepetae 80g, Radix Saposhnikoviae 80g, Pericarpium Citri Reticulatae Viride 100g, Cortex Moutan 100g, Radix Rehmanniae 150g, Indigo Naturalis 100g, Herba Asari 60g, Radix Angelicae Dahuricae 50g, Radix Glycyrrhizae 60g, Herba Schizonepetae, Herba Asari, the Radix Angelicae Dahuricae, Indigo Naturalis are joined to CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 30MPa, temperature 50 C, CO
2flow 3ml/g crude drug min, extraction time 180min, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 50% ethanol of 1.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600W, extracts 2 times, each 5 minutes, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 100, every heavy 0.8g.
After testing, in finished product, the content of Hesperidin is 76.2mg/ sheet.
Embodiment 3
Get Gypsum Fibrosum 200g, Herba Schizonepetae 80g, Radix Saposhnikoviae 80g, Pericarpium Citri Reticulatae Viride 100g, Cortex Moutan 100g, Radix Rehmanniae 150g, Indigo Naturalis 100g, Herba Asari 60g, Radix Angelicae Dahuricae 50g, Radix Glycyrrhizae 60g, Herba Schizonepetae, Herba Asari, the Radix Angelicae Dahuricae, Indigo Naturalis are joined to CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 15MPa, 30 ℃ of temperature, CO
2flow 1ml/g crude drug min, extraction time 220min, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 50% ethanol of 1.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 800W, extracts 2 times, each 10 minutes, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 100, every heavy 0.8g.
After testing, in finished product, the content of Hesperidin is 78.5mg/ sheet.
Embodiment 4: toothache Pianomide suppresses the experimentation data of mouse leukemia cell L1210 cell proliferation
1. experiment material
1.1 experiment cell strains
Mouse leukemia cell L1210 cell, Shandong Prov. Hospital laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: toothache Pianomide of the present invention: press embodiment 1 method preparation.
Medicinal liquid liquid storage: take 100mg toothache Pianomide, be dissolved in 5ml dehydrated alcohol, 0.2 m filter filters, 500 ldoff pipe packing ,-20 ℃ of storages, 0.2 m filter filters dehydrated alcohol in order to the use of matched group simultaneously.
1.3 experiment reagent
DMEM (the Cat.No.12100-061 Lot.No.758137 of GIBCO company); Hyclone (Lot.No.100419 of Tian Hang bio tech ltd, Zhejiang); NaHC0
3(the Cat.No.11810-033 Lot.No. of Shanghai Jiu Yi chemical reagent company limited 1088387); Trypsin(AMRESCO company 1); EDTA(AMRESCO company 1); Penicillin G Sodium Salt(AMRESCO company 1); Streptomycin Sulfate (AMRESCO); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.); MTT (Biosharp lot number: 0793): the autogamy of PBS(laboratory);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DMIL); Visible-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRAMAX 190); C0
2incubator (FORMA model: 3111); (safe and sound company of Su Jing group manufactures model to super-clean bench: SW-CJ-ZFD); Pure water instrument (U.S. Sprlng company 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 (Shanghai accurate experimental facilities company 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 m filter (MILLIPORE model: SLGP033RB); 1cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2. experimental technique
1) L1210 cell uses DMEM+10%FBS in 37 ℃, 5%C0
2carry out cellar culture (10cm culture dish), when Growth of Cells is during to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml 0.25% trypsin-0.04%EDTA, after 37 ℃ of digestion 2min, add wherein 5ml complete medium neutralization reaction, after piping and druming cell, proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjusts concentration of cell suspension 3 * 10
4individual/ml.
2) cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 l, culture plate put into cell culture incubator (37 ℃, 5%C0
2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add toothache Pianomide solution, continue to cultivate 24h.
4) after 24h, add 20 l MTT solution (5mg/ml, i.e. 0.5%MTT), continue to cultivate 4h.
5) after 4h, buckle method is removed supernatant, with absorbent paper, pats dry gently, and low-speed oscillation 10min on shaking table, as entered 200 l dimethyl sulfoxide, is put in every hole, and crystal is fully dissolved.At enzyme-linked immunosorbent assay instrument 490nm place, measure the light absorption value in each hole.
6) background (do not add cell, only add culture fluid) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide), sets 6 multiple holes for every group.
7) result represents the suppression ratio of cell with medicine: cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value), control wells OD value * 100%.Experiment repeats 3 times.
3. statistical disposition
Adopt correlation analysis and Student t check in Microsoft Excel 2007 softwares, data represent with mean ± S.D..
4. experimental result
Statistical result showed after mtt assay experiment, with matched group comparison, when dosage reaches 5mg/ml, to L1210 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has utmost point significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 toothache Pianomide is on L1210 cell inhibitory effect impact research (X ± SD)
Note: with matched group comparison, * P<O.01; * P<0.001.
5. experiment conclusion
Toothache Pianomide can suppress L1210 cell proliferation, reduces the Growth of Cells number of L1210 cell, and this effect is dose dependent.
Claims (9)
1. the preparation method of a toothache Pianomide, by Gypsum Fibrosum 200g, Herba Schizonepetae 80g, Radix Saposhnikoviae 80g, Pericarpium Citri Reticulatae Viride 100g, Cortex Moutan 100g, Radix Rehmanniae 150g, Indigo Naturalis 100g, Herba Asari 60g, Radix Angelicae Dahuricae 50g, Radix Glycyrrhizae 60g, as crude drug, made, it is characterized in that, described preparation method is comprised of the following step: get Herba Schizonepetae, Herba Asari, the Radix Angelicae Dahuricae, Indigo Naturalis, 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 ℃, CO
2flow l-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, standby; Get all the other Chinese medicines, pulverize, add 50% ethanol of 1.6L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power 600-800W, extracts 2 times, each 5-10 minute, combining extraction liquid, concentrated, be added on Dl01 macroporous adsorptive resins, 50% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, standby; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 100, every heavy 0.8g.
2. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that described CO
2in supercritical extraction, to account for the percent by volume of total extractant be 5% to entrainer.
3. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that described CO
2in supercritical extraction, extracting pressure is 25 MPa.
4. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that described CO
2in supercritical extraction, extraction temperature is 60 ℃.
5. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
6. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that described CO
2in supercritical extraction, extraction time is 200min.
7. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that, described microwave extracting power is 700W.
8. according to the preparation method of the toothache Pianomide described in claim l, it is characterized in that, the each extraction time of described microwave extracting is 8 minutes.
9. the toothache Pianomide described in claim l suppresses the application in mouse leukemia cell L1210 cell proliferation medicine in preparation.
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| CN106333989A (en) * | 2016-11-19 | 2017-01-18 | 张忠远 | Pharmaceutical composition for treating wind-heat toothache |
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