CN103028005A - Preparation method of anti-lumbago tablet and application thereof - Google Patents
Preparation method of anti-lumbago tablet and application thereof Download PDFInfo
- Publication number
- CN103028005A CN103028005A CN2012103784219A CN201210378421A CN103028005A CN 103028005 A CN103028005 A CN 103028005A CN 2012103784219 A CN2012103784219 A CN 2012103784219A CN 201210378421 A CN201210378421 A CN 201210378421A CN 103028005 A CN103028005 A CN 103028005A
- Authority
- CN
- China
- Prior art keywords
- preparation
- extraction
- lumbago
- lumbago tablet
- parched
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 208000008035 Back Pain Diseases 0.000 title claims abstract description 39
- 208000008930 Low Back Pain Diseases 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 239000003814 drug Substances 0.000 claims abstract description 20
- 150000003839 salts Chemical class 0.000 claims abstract description 14
- 239000004863 Frankincense Substances 0.000 claims abstract description 12
- 238000000874 microwave-assisted extraction Methods 0.000 claims abstract description 11
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 10
- 230000004663 cell proliferation Effects 0.000 claims abstract description 6
- 206010038389 Renal cancer Diseases 0.000 claims abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 55
- 238000000605 extraction Methods 0.000 claims description 19
- 239000000284 extract Substances 0.000 claims description 17
- 229940079593 drug Drugs 0.000 claims description 13
- 241000717739 Boswellia sacra Species 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 241001489978 Eupolyphaga Species 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 229920002472 Starch Polymers 0.000 claims description 5
- 230000000274 adsorptive effect Effects 0.000 claims description 5
- 229940126678 chinese medicines Drugs 0.000 claims description 5
- 230000006837 decompression Effects 0.000 claims description 5
- 239000003480 eluent Substances 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 5
- 238000004064 recycling Methods 0.000 claims description 5
- 239000011347 resin Substances 0.000 claims description 5
- 229920005989 resin Polymers 0.000 claims description 5
- 239000008107 starch Substances 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 208000006265 Renal cell carcinoma Diseases 0.000 claims description 4
- 201000010174 renal carcinoma Diseases 0.000 claims description 4
- 230000005764 inhibitory process Effects 0.000 claims description 3
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 abstract description 9
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 abstract description 9
- 229940114124 ferulic acid Drugs 0.000 abstract description 9
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 abstract description 9
- 235000001785 ferulic acid Nutrition 0.000 abstract description 9
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 abstract description 9
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 241000427159 Achyranthes Species 0.000 abstract 1
- 241000382455 Angelica sinensis Species 0.000 abstract 1
- 235000003717 Boswellia sacra Nutrition 0.000 abstract 1
- 240000007551 Boswellia serrata Species 0.000 abstract 1
- 235000012035 Boswellia serrata Nutrition 0.000 abstract 1
- 244000223760 Cinnamomum zeylanicum Species 0.000 abstract 1
- 241000123589 Dipsacus Species 0.000 abstract 1
- 208000008839 Kidney Neoplasms Diseases 0.000 abstract 1
- 244000170916 Paeonia officinalis Species 0.000 abstract 1
- 235000006484 Paeonia officinalis Nutrition 0.000 abstract 1
- 235000017803 cinnamon Nutrition 0.000 abstract 1
- 201000010982 kidney cancer Diseases 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 229960004756 ethanol Drugs 0.000 description 21
- 238000002474 experimental method Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 206010050031 Muscle strain Diseases 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- FCPVYOBCFFNJFS-LQDWTQKMSA-M benzylpenicillin sodium Chemical compound [Na+].N([C@H]1[C@H]2SC([C@@H](N2C1=O)C([O-])=O)(C)C)C(=O)CC1=CC=CC=C1 FCPVYOBCFFNJFS-LQDWTQKMSA-M 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012738 dissolution medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000004900 laundering Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- JWHHANVGNNWIRI-UHFFFAOYSA-N methanol phosphoric acid hydrate Chemical compound O.OC.OP(O)(O)=O JWHHANVGNNWIRI-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 229960002385 streptomycin sulfate Drugs 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Abstract
The invention provides a preparation method of an anti-lumbago tablet. The anti-lumbago tablet is prepared by using 108g of folium cortex eucommiae fried with salt, 81g of fructus psoraleae fried with salt, 81g of teasel root, 108g of angelica sinensis, 81g of fried rhizoma atractylodis macrocephalae, 81g of the root of bidentate achyranthes, 27g of cinnamon, 27g of prepared frankincense, 81g of prepared rhizoma cibotii, 43g of root of common peony, 54g of rhizoma alismatis and 43g of ground beeltle fried with wine as raw materials; and the anti-lumbago tablet is prepared by using supercritical extraction and microwave-assisted extraction, so that the content of ferulic acid is greatly enhanced. The invention also provides application of the anti-lumbago tablet in preparation of medicine for inhibiting cell proliferation of human kidney cancer cell KETR-3.
Description
Technical field
The present invention relates to the Chinese medicine preparation technical field, be specifically related to a kind of preparation method and application of anti-lumbago tablet.
Background technology
Anti-lumbago tablet is recorded in Ministry of Public Health standard WS3-B-1053-91, made as crude drug by Cortex Eucommiae (parched with salt) leaf 108g, salt Fructus Psoraleae (parched) 81g, Radix Dipsaci 81g, Radix Angelicae Sinensis 108g, Rhizoma Atractylodis Macrocephalae (parched) 81g, Radix Achyranthis Bidentatae 81g, Cortex Cinnamomi 27g, Olibanum (processed) 27g, Rhizoma Cibotii 81g processed, Radix Paeoniae Rubra 43g, Rhizoma Alismatis 54g, wine Eupolyphaga (parched) 43g, the energy strengthening vital energy of kidney, promoting blood circulation and stopping pain.Be used for lumbago due to renal deficiency, lumbar muscle strain.
In the prior art, not yet there is anti-lumbago tablet to adopt the report of supercritical and microwave technology aspect the preparation extracting, and adopts the method that powder and decocting boil of beating, 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 used clinically.
Summary of the invention
Goal of the invention: in order to address the above problem, the object of the present invention is to provide a kind of preparation method of anti-lumbago tablet.
Another object of the present invention is to provide the application of a kind of anti-lumbago tablet in preparation inhibition human renal carcinoma cell KETR-3 cell proliferation medicine.
Technical scheme: the objective of the invention is to realize by following scheme:
A kind of preparation method of anti-lumbago tablet, by Cortex Eucommiae (parched with salt) leaf 108g, salt Fructus Psoraleae (parched) 81g, Radix Dipsaci 81g, Radix Angelicae Sinensis 108g, Rhizoma Atractylodis Macrocephalae (parched) 81g, Radix Achyranthis Bidentatae 81g, Cortex Cinnamomi 27g, Olibanum (processed) 27g, Rhizoma Cibotii 81g processed, Radix Paeoniae Rubra 43g, Rhizoma Alismatis 54g, wine Eupolyphaga (parched) 43g makes as crude drug, described method is comprised of the following step: get Radix Angelicae Sinensis, Cortex Cinnamomi, Olibanum (processed), join in the 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min gets supercritical extract, and is for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in the microwave extracting apparatus and carry out microwave extracting, extraction power 400-600W extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on the D101 macroporous adsorptive resins, 50% ethanol elution is collected 5 times of amount column volume eluents, decompression recycling ethanol, concentrated and dry, get the microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, drying, tabletting is made 1000, every heavy 0.5g.
The preparation method of above-mentioned a kind of anti-lumbago tablet, described CO
2The percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned a kind of anti-lumbago tablet, described microwave extracting power 500W extracts 6 minutes at every turn.
The preparation method of above-mentioned a kind of anti-lumbago tablet, described CO
2The extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2Flow 2ml/g crude drug min, extraction time 160min.
The application of above-mentioned anti-lumbago tablet in preparation inhibition human renal carcinoma cell KETR-3 cell proliferation medicine.
In the prior art, every 0.5g of anti-lumbago tablet, each 6,3 times on the one, the every 0.5g of anti-lumbago tablet that adopts the present invention to be prepared into only needs 3 at every turn, takes 3 times in 1st, has greatly reduced dose having under the condition of more active component.This conclusion can be by following evidence.
The comparison of ferulaic acid content in the anti-lumbago tablet of test one, distinct methods preparation
1, instrument and reagent anti-lumbago tablet of the present invention: press the preparation of embodiment 3 methods, use the 815g crude drug, make 1000 through extraction, every heavy 0.5g.Former anti-lumbago tablet by the preparation of ministry standard method, uses the 815g crude drug, makes 1000 through extraction, every heavy 0.5g.Agilent 1200 high performance liquid chromatographs; METTLER AE240 electronic analytical balance; Ferulic acid reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute).
2, method
Chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica; Methanol-water-phosphoric acid (40:60:0.2) is mobile phase; The detection wavelength is 280nm.Number of theoretical plate is pressed the ferulic acid peak and is calculated, and should be not less than 3000.
The preparation of reference substance solution: precision takes by weighing at 4 hours ferulic acid reference substance of 60 ℃ of drying under reduced pressure an amount of, adds methanol and makes the solution that every 1ml contains 18 μ g, and get final product.
The preparation of need testing solution: get anti-lumbago tablet of the present invention and former anti-lumbago tablet, porphyrize, mixing is got 1g, and is accurately weighed, the accurate 70% ethanol 20ml that adds, close plug, supersound process 10 minutes, centrifugal, get supernatant, and get final product.
Algoscopy is accurate reference substance solution and each 20 μ l of need testing solution of drawing respectively, and the injection liquid chromatography is measured, and be get final product.
3, result
The result shows that the content of ferulic acid is the 1.12mg/ sheet in the anti-lumbago tablet of the present invention; And the content of ferulic acid is the 0.14mg/ sheet in the former anti-lumbago tablet, and in the situation that dose reduces, ferulaic acid content improves a lot.
Above-mentioned studies show that, the anti-lumbago tablet that adopts the present invention to prepare, active constituent content is higher than the standby anti-lumbago tablet of ministry standard legal system.
The specific embodiment
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 only limits to following example, all technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Cortex Eucommiae (parched with salt) leaf 108g, salt Fructus Psoraleae (parched) 81g, Radix Dipsaci 81g, Radix Angelicae Sinensis 108g, Rhizoma Atractylodis Macrocephalae (parched) 81g, Radix Achyranthis Bidentatae 81g, Cortex Cinnamomi 27g, Olibanum (processed) 27g, Rhizoma Cibotii 81g processed, Radix Paeoniae Rubra 43g, Rhizoma Alismatis 54g, wine Eupolyphaga (parched) 43g, with Radix Angelicae Sinensis, Cortex Cinnamomi, Olibanum (processed), join in the CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, 30 ℃ of temperature, CO2 flow 1m1/g crude drug min, extraction time 150min, get supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in the microwave extracting apparatus and carry out microwave extracting, extraction power 400W extracts 2 times, each 4 minutes, combining extraction liquid, concentrated, be added on the D101 macroporous adsorptive resins, 50% ethanol elution is collected 5 times of amount column volume eluents, decompression recycling ethanol, concentrated and dry, get the microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, drying, tabletting is made 1000, every heavy 0.5g.
After testing, the content of ferulic acid is the 1.15mg/ sheet in the finished product.
Embodiment 2
Get Cortex Eucommiae (parched with salt) leaf 108g, salt Fructus Psoraleae (parched) 81g, Radix Dipsaci 81g, Radix Angelicae Sinensis 108g, Rhizoma Atractylodis Macrocephalae (parched) 81g, Radix Achyranthis Bidentatae 81g, Cortex Cinnamomi 27g, Olibanum (processed) 27g, Rhizoma Cibotii 81g processed, Radix Paeoniae Rubra 43g, Rhizoma Alismatis 54g, wine Eupolyphaga (parched) 43g, with Radix Angelicae Sinensis, Cortex Cinnamomi, Olibanum (processed), join CO
2In the supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa, temperature 50 C, CO
2Flow 3m1/g crude drug min, extraction time 180min gets supercritical extract, and is for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in the microwave extracting apparatus and carry out microwave extracting, extraction power 600W extracts 2 times, each 8 minutes, combining extraction liquid, concentrated, be added on the D101 macroporous adsorptive resins, 50% ethanol elution is collected 5 times of amount column volume eluents, decompression recycling ethanol, concentrated and dry, get the microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, drying, tabletting is made 1000, every heavy 0.5g.
After testing, the content of ferulic acid is the 1.09mg/ sheet in the finished product.
Embodiment 3
Get Cortex Eucommiae (parched with salt) leaf 108g, salt Fructus Psoraleae (parched) 81g, Radix Dipsaci 81g, Radix Angelicae Sinensis 108g, Rhizoma Atractylodis Macrocephalae (parched) 81g, Radix Achyranthis Bidentatae 81g, Cortex Cinnamomi 27g, Olibanum (processed) 27g, Rhizoma Cibotii 81g processed, Radix Paeoniae Rubra 43g, Rhizoma Alismatis 54g, wine Eupolyphaga (parched) 43g, with Radix Angelicae Sinensis, Cortex Cinnamomi, Olibanum (processed), join CO
2In the supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, 40 ℃ of temperature, CO
2Flow 2m1/g crude drug min, extraction time 160min gets supercritical extract, and is for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in the microwave extracting apparatus and carry out microwave extracting, extraction power 500W extracts 2 times, each 6 minutes, combining extraction liquid, concentrated, be added on the D101 macroporous adsorptive resins, 50% ethanol elution is collected 5 times of amount column volume eluents, decompression recycling ethanol, concentrated and dry, get the microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, drying, tabletting is made 1000, every heavy 0.5g.
After testing, the content of ferulic acid is the 1.12mg/ sheet in the finished product.
Embodiment 4: anti-lumbago tablet suppresses the experimentation data of KETR-3 cell proliferation
1 experiment material
1.1 experiment cell strain
Human renal carcinoma cell KETR-3 cell, Nanjing Zhengkuan Pharmaceutical Technology Co., Ltd.'s laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: anti-lumbago tablet of the present invention: press the preparation of embodiment 3 methods.
The medicinal liquid liquid storage: take by weighing the 100mg anti-lumbago tablet, be dissolved in the 5ml dehydrated alcohol, 0.2 μ m filter filters, and 500 μ l doff manage packing ,-20 ℃ of storages, and 0.2 μ m filter filters dehydrated alcohol in order to the usefulness of matched group simultaneously.
1.3 experiment reagent
The Cat.No.12100-061 Lot.No.758137 of DMEM(GIBCO company); Hyclone (Hangzhoupro, sky, Zhejiang bio tech ltd 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); The autogamy of PBS(laboratory);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); As seen-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRAMAX 190); CO2 incubator (FORMA model: 3111); (safe and sound company of Su Jing group makes model to super-clean bench: SW-CJ-ZFD); Pure water instrument (U.S. Spring 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 (the accurate experimental facilities in Shanghai 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 μ m 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) the KETR-3 cell carries out cellar culture (10cm culture dish) with DMEM+10%FBS in 37 ℃, 5%CO2, when Growth of Cells during to logarithmic (log) phase, collecting cell discards culture fluid, PBS fine laundering 3 times, add 3ml 0.25% trypsin-0.04%EDTA, behind 37 ℃ of digestion 2min, to wherein adding 5ml complete medium neutralization reaction, behind the piping and druming cell it is changed in the centrifuge tube, the centrifugal 5min of 1000rpm adjusts 3 * 104/ml of concentration of cell suspension.
2) the cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 ℃, 5%CO2) cellar culture.
3) according to the Growth of Cells situation, generally grow to 50%-70%, add anti-lumbago tablet solution, continue to cultivate 24h.
4) add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT) behind the 24h, continue to cultivate 4h.
5) the buckle method is removed supernatant behind the 4h, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on the shaking table, and crystal is fully dissolved.Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place.
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) is set 6 multiple holes for every group.
7) result represents with the suppression ratio of medicine to cell:
Cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value * 100%.Experiment repeats 3 times.
3 statistical dispositions
Adopt correlation analysis and Student t check in Microsoft Excel 2003 softwares, data represent with mean ± S.D..
4 experimental results
Statistical result showed after the mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to KETR-3 cell inhibitory effect variant (P＜0.05), dosage this difference when 10mg/ml has significance (P＜0.01), and utmost point significant difference (P＜0.001) is arranged when dosage reaches 15-20mg/ml.
Table 1 anti-lumbago tablet is on KETR-3 cell inhibitory effect impact research
Annotate: compare * P＜0.01 with matched group; * P＜0.001
5 experiment conclusion
Anti-lumbago tablet can suppress the KETR-3 cell proliferation, reduces the Growth of Cells number of KETR-3 cell, and this effect is dose dependent.
Claims (5)
1. the preparation method of an anti-lumbago tablet, made as crude drug by Cortex Eucommiae (parched with salt) leaf 108g, salt Fructus Psoraleae (parched) 81g, Radix Dipsaci 81g, Radix Angelicae Sinensis 108g, Rhizoma Atractylodis Macrocephalae (parched) 81g, Radix Achyranthis Bidentatae 81g, Cortex Cinnamomi 27g, Olibanum (processed) 27g, Rhizoma Cibotii 81g processed, Radix Paeoniae Rubra 43g, Rhizoma Alismatis 54g, wine Eupolyphaga (parched) 43g, it is characterized in that described method is comprised of the following step: get Radix Angelicae Sinensis, Cortex Cinnamomi, Olibanum (processed), join CO
2In the 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 1-3m1/g crude drug min, extraction time 150-180min gets supercritical extract, and is for subsequent use; Get all the other Chinese medicines, pulverize, add 70% ethanol of 2L, drop in the microwave extracting apparatus and carry out microwave extracting, extraction power 400-600W extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on the D101 macroporous adsorptive resins, 50% ethanol elution is collected 5 times of amount column volume eluents, decompression recycling ethanol, concentrated and dry, get the microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, drying, tabletting is made 1000, every heavy 0.5g.
2. the preparation method of described a kind of anti-lumbago tablet according to claim 1 is characterized in that described CO
2The percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
3. the preparation method of described a kind of anti-lumbago tablet according to claim 1 is characterized in that described microwave extracting power 500W, extracts 6 minutes at every turn.
4. the preparation method of described a kind of anti-lumbago tablet according to claim 1 is characterized in that described CO
2The extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2Flow 2ml/g crude drug min, extraction time 160min.
5. the according to claim 1 application of described a kind of anti-lumbago tablet in preparation inhibition human renal carcinoma cell KETR-3 cell proliferation medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210378421.9A CN103028005B (en) | 2012-10-08 | 2012-10-08 | Preparation method of anti-lumbago tablet and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210378421.9A CN103028005B (en) | 2012-10-08 | 2012-10-08 | Preparation method of anti-lumbago tablet and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103028005A true CN103028005A (en) | 2013-04-10 |
| CN103028005B CN103028005B (en) | 2014-06-11 |
Family
ID=48015734
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210378421.9A Expired - Fee Related CN103028005B (en) | 2012-10-08 | 2012-10-08 | Preparation method of anti-lumbago tablet and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103028005B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103638322A (en) * | 2013-11-28 | 2014-03-19 | 常州科立信医疗器械有限公司 | Preparation method and application of kidney-harmonizing pill |
| CN103638489A (en) * | 2013-12-04 | 2014-03-19 | 常州科立信医疗器械有限公司 | Preparation method and application of kidney-tonifying pills |
| CN103705624A (en) * | 2013-12-22 | 2014-04-09 | 青岛琴诚医药技术有限公司 | Preparation method and application of life-prolonging tablet |
| CN104173629A (en) * | 2014-08-16 | 2014-12-03 | 黑龙江江恒医药科技有限公司 | Anti-lumbago tablet and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1748753A (en) * | 2005-10-20 | 2006-03-22 | 北京阜康仁生物制药科技有限公司 | Waist pain preparation and new preparing method |
| CN102085280A (en) * | 2009-12-06 | 2011-06-08 | 湖北广仁药业有限公司 | One-step granulating method of tablets for treating lumbago |
-
2012
- 2012-10-08 CN CN201210378421.9A patent/CN103028005B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1748753A (en) * | 2005-10-20 | 2006-03-22 | 北京阜康仁生物制药科技有限公司 | Waist pain preparation and new preparing method |
| CN102085280A (en) * | 2009-12-06 | 2011-06-08 | 湖北广仁药业有限公司 | One-step granulating method of tablets for treating lumbago |
Non-Patent Citations (1)
| Title |
|---|
| 李攻科等: "微波辅助萃取技术在中药现代化中的应用", 《精细化工》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103638322A (en) * | 2013-11-28 | 2014-03-19 | 常州科立信医疗器械有限公司 | Preparation method and application of kidney-harmonizing pill |
| CN103638489A (en) * | 2013-12-04 | 2014-03-19 | 常州科立信医疗器械有限公司 | Preparation method and application of kidney-tonifying pills |
| CN103638489B (en) * | 2013-12-04 | 2015-07-15 | 新乡医学院第一附属医院 | Preparation method and application of kidney-tonifying pills |
| CN103705624A (en) * | 2013-12-22 | 2014-04-09 | 青岛琴诚医药技术有限公司 | Preparation method and application of life-prolonging tablet |
| CN104173629A (en) * | 2014-08-16 | 2014-12-03 | 黑龙江江恒医药科技有限公司 | Anti-lumbago tablet and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103028005B (en) | 2014-06-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102988723B (en) | Preparation method and application of traditional Chinese medicine | |
| CN102988526B (en) | Method for preparing Bailing tablets and application | |
| CN102988667B (en) | Preparation method and application of tablet for regulating menstruation and activating blood | |
| CN102988577B (en) | Preparation method and application of brain-soothing hypertension pill | |
| CN102988665B (en) | Method for preparing gynecological menstruation regulating tablets and application | |
| CN102988612B (en) | Preparation method and application of tablet for clearing away heat | |
| CN103028005B (en) | Preparation method of anti-lumbago tablet and application thereof | |
| CN102988502B (en) | Preparation method and application of Desheng tablet | |
| CN102988545B (en) | Preparation method and application of Yulian tablet | |
| CN102988489B (en) | Preparation method and application of nose comforting tablet | |
| CN102988480B (en) | Method for preparing bile and patchouli rhinitis tablets and application | |
| CN102988479B (en) | Preparation method and application of anti-dazzling tablet | |
| CN102988550B (en) | Preparation method and application of qinlian tablet | |
| CN102988509B (en) | Preparation method and application of bone-knitting tablet | |
| CN102836381A (en) | Preparation method and application of middle calming tablet | |
| CN102988614A (en) | Preparation method and application of antiphlogosis tablet | |
| CN102988881B (en) | Preparation method and application of pulse unblocking tablet | |
| CN102988611B (en) | Preparation method and application of antihypertension tablet | |
| CN102973749A (en) | Preparation method and applications of divaricate saposhnikovia-angelica archang lica rhinitis tablets | |
| CN102988500B (en) | Method for preparing compound Dantong tablets and application | |
| CN102988748B (en) | Preparation method and application of yin tonifying and blood sugar reducing tablet | |
| CN102988522B (en) | Preparation method and application of naodesheng tablets | |
| CN102988501A (en) | Preparation method and application of Libiling tablet | |
| CN102988749B (en) | Preparation method and application of body-building tablet | |
| CN102988691B (en) | Preparation method and application of liver-strengthening tablet |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| ASS | Succession or assignment of patent right |
Owner name: MAO JIE Free format text: FORMER OWNER: BIAN YUPING Effective date: 20140506 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| C53 | Correction of patent of invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Zhang Peng Inventor after: Mao Jie Inventor before: The inventor has waived the right to be mentioned |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: REQUEST NOT TO RELEASE THE NAME TO: ZHANG PENG MAO JIE Free format text: CORRECT: ADDRESS; FROM: 211224 NANJING, JIANGSU PROVINCE TO: 315041 NINGBO, ZHEJIANG PROVINCE |
|
| TA01 | Transfer of patent application right |
Effective date of registration: 20140506 Address after: 315041 No. 456, Xingning Road, Jiangdong District, Zhejiang, Ningbo Applicant after: Mao Jie Address before: Lishui County of Nanjing City, Jiangsu province 211224 Jing Qiao Zhen Chen Biancun Applicant before: Bian Yuping |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee | ||
| CP02 | Change in the address of a patent holder |
Address after: 225816, No. 5, Christmas road, industrial town, Baoying County, Jiangsu, Yangzhou Patentee after: Mao Jie Address before: 315041 No. 456, Xingning Road, Jiangdong District, Zhejiang, Ningbo Patentee before: Mao Jie |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| CB03 | Change of inventor or designer information |
Inventor after: Lv Hongfeng Inventor before: Zhang Peng Inventor before: Mao Jie |
|
| COR | Change of bibliographic data | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160701 Address after: 262500 Shandong city of Weifang province Qingzhou City, No. 391 Zheng Mu Zhen Patentee after: Lv Hongfeng Address before: 225816, No. 5, Christmas road, industrial town, Baoying County, Jiangsu, Yangzhou Patentee before: Mao Jie |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140611 Termination date: 20161008 |