CN104116810A - Preparation method of Jiuwei haemorrhoids tablet and application of Jiuwei haemorrhoids tablet to medicine for inhibiting cell proliferation of lymphoma cell YAC-1 - Google Patents
Preparation method of Jiuwei haemorrhoids tablet and application of Jiuwei haemorrhoids tablet to medicine for inhibiting cell proliferation of lymphoma cell YAC-1 Download PDFInfo
- Publication number
- CN104116810A CN104116810A CN201410371536.4A CN201410371536A CN104116810A CN 104116810 A CN104116810 A CN 104116810A CN 201410371536 A CN201410371536 A CN 201410371536A CN 104116810 A CN104116810 A CN 104116810A
- Authority
- CN
- China
- Prior art keywords
- preparation
- extraction
- medicinal herbs
- jiuwei
- haemorrhoids
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 239000003814 drug Substances 0.000 title claims abstract description 23
- 101000981253 Mus musculus GPI-linked NAD(P)(+)-arginine ADP-ribosyltransferase 1 Proteins 0.000 title claims abstract description 14
- 206010025323 Lymphomas Diseases 0.000 title claims abstract description 9
- 230000004663 cell proliferation Effects 0.000 title claims abstract description 9
- 208000014617 hemorrhoid Diseases 0.000 title abstract description 40
- 230000002401 inhibitory effect Effects 0.000 title abstract description 4
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 16
- 229940079593 drug Drugs 0.000 claims abstract description 14
- 238000000874 microwave-assisted extraction Methods 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 53
- 235000008216 herbs Nutrition 0.000 claims description 42
- 210000004027 cell Anatomy 0.000 claims description 22
- 238000000605 extraction Methods 0.000 claims description 22
- 239000000284 extract Substances 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 13
- 102000002322 Egg Proteins Human genes 0.000 claims description 6
- 108010000912 Egg Proteins Proteins 0.000 claims description 6
- 235000008730 Ficus carica Nutrition 0.000 claims description 6
- 244000025361 Ficus carica Species 0.000 claims description 6
- 241000628997 Flos Species 0.000 claims description 6
- 241000287828 Gallus gallus Species 0.000 claims description 6
- 229940126678 chinese medicines Drugs 0.000 claims description 6
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 6
- 210000004681 ovum Anatomy 0.000 claims description 6
- 229920002472 Starch Polymers 0.000 claims description 5
- 230000000274 adsorptive effect Effects 0.000 claims description 5
- 239000012141 concentrate Substances 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
- 229920005989 resin Polymers 0.000 claims description 5
- 239000011347 resin Substances 0.000 claims description 5
- 239000008107 starch Substances 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 239000010282 Emodin Substances 0.000 abstract description 12
- RHMXXJGYXNZAPX-UHFFFAOYSA-N emodin Chemical compound C1=C(O)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O RHMXXJGYXNZAPX-UHFFFAOYSA-N 0.000 abstract description 12
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 2
- 235000007516 Chrysanthemum Nutrition 0.000 abstract 1
- 244000189548 Chrysanthemum x morifolium Species 0.000 abstract 1
- 241000037740 Coptis chinensis Species 0.000 abstract 1
- 240000004064 Poterium sanguisorba Species 0.000 abstract 1
- 235000008291 Poterium sanguisorba Nutrition 0.000 abstract 1
- 240000001341 Reynoutria japonica Species 0.000 abstract 1
- 235000018167 Reynoutria japonica Nutrition 0.000 abstract 1
- 240000004980 Rheum officinale Species 0.000 abstract 1
- 235000008081 Rheum officinale Nutrition 0.000 abstract 1
- 240000005255 Rubus parvifolius Species 0.000 abstract 1
- 235000018803 Rubus parvifolius Nutrition 0.000 abstract 1
- 241000124033 Salix Species 0.000 abstract 1
- 235000008282 Sanguisorba officinalis Nutrition 0.000 abstract 1
- 230000024241 parasitism Effects 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 27
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 24
- 229960004756 ethanol Drugs 0.000 description 20
- VWDXGKUTGQJJHJ-UHFFFAOYSA-N Catenarin Natural products C1=C(O)C=C2C(=O)C3=C(O)C(C)=CC(O)=C3C(=O)C2=C1O VWDXGKUTGQJJHJ-UHFFFAOYSA-N 0.000 description 11
- RBLJKYCRSCQLRP-UHFFFAOYSA-N Emodin-dianthron Natural products O=C1C2=CC(C)=CC(O)=C2C(=O)C2=C1CC(=O)C=C2O RBLJKYCRSCQLRP-UHFFFAOYSA-N 0.000 description 11
- YOOXNSPYGCZLAX-UHFFFAOYSA-N Helminthosporin Natural products C1=CC(O)=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O YOOXNSPYGCZLAX-UHFFFAOYSA-N 0.000 description 11
- NTGIIKCGBNGQAR-UHFFFAOYSA-N Rheoemodin Natural products C1=C(O)C=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1O NTGIIKCGBNGQAR-UHFFFAOYSA-N 0.000 description 11
- VASFLQKDXBAWEL-UHFFFAOYSA-N emodin Natural products OC1=C(OC2=C(C=CC(=C2C1=O)O)O)C1=CC=C(C=C1)O VASFLQKDXBAWEL-UHFFFAOYSA-N 0.000 description 11
- PKUBGLYEOAJPEG-UHFFFAOYSA-N physcion Natural products C1=C(C)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O PKUBGLYEOAJPEG-UHFFFAOYSA-N 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 239000002775 capsule Substances 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 8
- 238000012360 testing method Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000001629 suppression Effects 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
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 241000167880 Hirundinidae Species 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
- 238000009825 accumulation Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000010165 autogamy Effects 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
- 235000019658 bitter taste Nutrition 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 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
- 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
- 239000000945 filler Substances 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 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
- 238000007689 inspection Methods 0.000 description 1
- 201000007772 internal hemorrhoid Diseases 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
- 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
- 231100000614 poison Toxicity 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 229960002385 streptomycin sulfate Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a preparation method of a Jiuwei haemorrhoids tablet and an application of the Jiuwei haemorrhoids tablet. The Jiuwei haemorrhoids tablet is prepared by 400g of rubus parvifolius, 150g of polygonum cuspidatum, 200g of garden burnet, 100g of willow parasitism, 100g of fig leaf, 50g of chrysanthemum, 20g of hen egg-albumen, 150g of coptis chinensis and 50g of rheum officinale serving as raw drugs via supercritical extraction and microwave extraction, so that the content of rheum emodin is greatly improved. The invention also provides an application of the Jiuwei haemorrhoids tablet to a medicine for inhibiting cell proliferation of mouse lymphoma cell YAC-1.
Description
Technical field
The invention belongs to technical field of Chinese medicines, be specifically related to a kind of preparation method for the treatment of piles with 9-medicinal herbs sheet and the application in inhibition mouse lymphoma cell YAC-1 cell proliferation medicine thereof.
Background technology
Capsule for treating piles with 9-medicinal herbs standard No. WS-10189 (ZD-0189)-2002, is recorded in national standard for traditional Chinese medicines compilation surgery gynecological fascicle.Made as crude drug by Herba Rubi parvifolii 400g, Rhizoma Polygoni Cuspidati 150g, Radix Sanguisorbae 200g, Liu Jisheng 100g, common fig leaf juice 100g, Flos Chrysanthemi 50g, Ovum Gallus domesticus album 20g, Rhizoma Coptidis 150g, Radix Et Rhizoma Rhei 50g, there is heat-clearing and toxic substances removing, dampness detumescence, effect of cooling blood for hemostasis.。For Internal hemorrhoid hemorrhage due to damp-heat accumulation, external hemorrhoid welling and pain.
In prior art, not yet there is capsule for treating piles with 9-medicinal herbs adopting the report of supercritical and microwave technology aspect extraction preparation, and directly beat powder, the method that decocting boils, technique is coarse, backward, and impurity is many, cause patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
In prior art, the each 5-6 grain of capsule for treating piles with 9-medicinal herbs, 3 times on the one.Capsule for treating piles with 9-medicinal herbs dosage is large, and the every heavy 0.3g for the treatment of piles with 9-medicinal herbs sheet that adopts the inventive method to be prepared into only needs 3 at every turn, within 1st, takes 3 times.Under the condition with more active component, greatly reduce dose.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 emodin content in treating piles with 9-medicinal herbs sheet prepared by test one, distinct methods
L, instrument and reagent treating piles with 9-medicinal herbs sheet of the present invention: press embodiment 1 method preparation, use 1220g crude drug, make 500 through extracting, every heavy 0.3g.Former capsule for treating piles with 9-medicinal herbs, prepares according to WS-10189 (ZD-0189)-2002 standard method.Agilent1200 high performance liquid chromatograph; METTLER AE240 electronic analytical balance; Emodin reference substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
Measure according to high performance liquid chromatography (annex VI D of Chinese Pharmacopoeia version in 2010).
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are filler; Methanol-0.1% phosphoric acid solution (80:20) is mobile phase; Detection wavelength is 288nm.Number of theoretical plate calculates and should be not less than 5000 by emodin peak.
It is appropriate that the preparation precision of reference substance solution takes emodin reference substance, adds methanol and make the solution of every 1ml containing 0.05mg, to obtain final product.
Treating piles with 9-medicinal herbs sheet of the present invention is got in the preparation of need testing solution, and porphyrize, gets 0.5g, accurately weighed, put in tool plug conical flask, add methanol 20ml, supersound process 1 hour, lets cool, filter residue methanol wash 2 times, each 10ml, merging filtrate and washing liquid, evaporate to dryness, residue adds dilute hydrochloric acid 10ml to be made to dissolve, add again chloroform 20ml, put (70 DEG C) in water-bath and be hydrolyzed 30 minutes, cooling, in dislocation separatory funnel, with a small amount of chloroform washing container, be incorporated in separatory funnel; Point get chloroform layer, chloroform washing 2 times for acid solution, each 10ml, combined chloroform liquid, dewater in right amount with anhydrous sodium sulfate, fling to chloroform, residue makes to dissolve and be transferred in 10ml measuring bottle with methanol in right amount, adds methanol to scale, shake up, with microporous filter membrane, (0.45 m) filtration, gets subsequent filtrate, to obtain final product.
The capsule for treating piles with 9-medicinal herbs of this contrast is got in the preparation of reference product need testing solution, and porphyrize mixes, and gets 1g, accurately weighed, put in tool plug conical flask, add methanol 20ml, supersound process 1 hour, let cool, filter, residue methanol wash 2 times, each 10ml, merging filtrate and washing liquid, evaporate to dryness, residue adds dilute hydrochloric acid 10ml to be made to dissolve, add again chloroform 20ml, put (70 DEG C) in water-bath and be hydrolyzed 30 minutes, cooling, in dislocation separatory funnel, with a small amount of chloroform washing container, be incorporated in separatory funnel; Point get chloroform layer, chloroform washing 2 times for acid solution, each 10ml, combined chloroform liquid, dewater in right amount with anhydrous sodium sulfate, fling to chloroform, residue makes to dissolve and be transferred in 10ml measuring bottle with methanol in right amount, adds methanol to scale, shake up, with microporous filter membrane, (0.45 m) filtration, gets subsequent filtrate, to obtain final product.
Algoscopy is accurate reference substance solution and each 5 l of need testing solution of drawing respectively, and injection liquid chromatography, measures, and to obtain final product.3, result
Result shows, in treating piles with 9-medicinal herbs sheet of the present invention, the content of emodin is 0.65-1.22mg/ sheet; And the content of emodin is 0.15mg/ grain in former capsule for treating piles with 9-medicinal herbs, be 2-4 times of original capsule content each serving the emodin content of 3 of consumptions, in the situation that dose reduces, emodin content improves a lot.
Above-mentioned research shows, the treating piles with 9-medicinal herbs sheet that adopts preparation method of the present invention to prepare, capsule for treating piles with 9-medicinal herbs prepared by the method that active constituent content is recorded higher than WS-10189 (ZD-0189)-2002 standard far away.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides a kind of preparation method for the treatment of piles with 9-medicinal herbs sheet.
Another object of the present invention is to provide a kind for the treatment of piles with 9-medicinal herbs sheet to suppress the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
The object of the invention is to realize by following scheme:
A kind of preparation method for the treatment of piles with 9-medicinal herbs sheet, made as crude drug by Herba Rubi parvifolii 400g, Rhizoma Polygoni Cuspidati 150g, Radix Sanguisorbae 200g, Liu Jisheng 100g, common fig leaf juice 100g, Flos Chrysanthemi 50g, Ovum Gallus domesticus album 20g, Rhizoma Coptidis 150g, Radix Et Rhizoma Rhei 50g, described preparation method is made up of the following step: get Herba Rubi parvifolii, Rhizoma Polygoni Cuspidati, Liu Jisheng, 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 l-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 60% ethanol of 5L, 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, 60% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.3g.
In the preparation method of above-mentioned treating piles with 9-medicinal herbs sheet, 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 treating piles with 9-medicinal herbs sheet, described CO
2in supercritical extraction, extracting pressure is 25 MPa.
In the preparation method of above-mentioned treating piles with 9-medicinal herbs sheet, described CO
2in supercritical extraction, extraction temperature is 40 DEG C.
In the preparation method of above-mentioned treating piles with 9-medicinal herbs sheet, described CO
2cO in supercritical extraction
2flow 2ml/g crude drug min.
In the preparation method of above-mentioned treating piles with 9-medicinal herbs sheet, described CO
2in supercritical extraction, extraction time is 200min.
In the preparation method of above-mentioned treating piles with 9-medicinal herbs sheet, described microwave extracting power is 700W.
In the preparation method of above-mentioned treating piles with 9-medicinal herbs sheet, the each extraction time of described microwave extracting is 8 minutes.
Above-mentioned treating piles with 9-medicinal herbs sheet suppresses the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
Detailed description of the invention
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 Herba Rubi parvifolii 400g, Rhizoma Polygoni Cuspidati 150g, Radix Sanguisorbae 200g, Liu Jisheng 100g, common fig leaf juice 100g, Flos Chrysanthemi 50g, Ovum Gallus domesticus album 20g, Rhizoma Coptidis 150g, Radix Et Rhizoma Rhei 50g, Herba Rubi parvifolii, Rhizoma Polygoni Cuspidati, Liu Jisheng 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 DEG C of temperature, CO
2flow 2ml/g crude drug min, extraction time 200min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 60% ethanol of 5L, 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, 60% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.3g.
After testing, in finished product, the content of emodin is 1.22mg/ sheet.
Embodiment 2
Get Herba Rubi parvifolii 400g, Rhizoma Polygoni Cuspidati 150g, Radix Sanguisorbae 200g, Liu Jisheng 100g, common fig leaf juice 100g, Flos Chrysanthemi 50g, Ovum Gallus domesticus album 20g, Rhizoma Coptidis 150g, Radix Et Rhizoma Rhei 50g, Herba Rubi parvifolii, Rhizoma Polygoni Cuspidati, Liu Jisheng 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, for subsequent use; Get all the other Chinese medicines, pulverize, add 60% ethanol of 5L, 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, 60% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.3g.
After testing, in finished product, the content of emodin is 0.87mg/ sheet.
Embodiment 3
Get Herba Rubi parvifolii 400g, Rhizoma Polygoni Cuspidati 150g, Radix Sanguisorbae 200g, Liu Jisheng 100g, common fig leaf juice 100g, Flos Chrysanthemi 50g, Ovum Gallus domesticus album 20g, Rhizoma Coptidis 150g, Radix Et Rhizoma Rhei 50g, Herba Rubi parvifolii, Rhizoma Polygoni Cuspidati, Liu Jisheng 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 DEG C of temperature, CO
2flow 1ml/g crude drug min, extraction time 220min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 60% ethanol of 5L, 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, 60% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.3g.
After testing, in finished product, the content of emodin is 0.65mg/ sheet.
Embodiment 4: treating piles with 9-medicinal herbs sheet suppresses the experimentation data of mouse lymphoma cell YAC-1 cell proliferation
1. experiment material
1.1 experiment cell strains
Mouse lymphoma cell YAC-1 cell, Shandong University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: treating piles with 9-medicinal herbs sheet of the present invention: press embodiment 1 method preparation.
Medicinal liquid liquid storage: take 100mg treating piles with 9-medicinal herbs sheet, be dissolved in 5ml dehydrated alcohol, 0.2 m filter filters, 500 ldoff pipe subpackages ,-20 DEG C of storages, 0.2 m filter filters the use of dehydrated alcohol in order to 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 Shanghai hundred million Cat.No.11810-033 Lot.No. of chemical reagent company limited 1088387 of a specified duration); Trypsin(AMRESCO company); EDTA(AMRESCO company); Penicillin G Sodium Salt(AMRESCO company 1); Streptomycin Sulfate (AMRESCO); Dehydrated alcohol (Zibo Ya Dulan Trade Co., Ltd.); MTT (Biosharp lot number: 0793): PBS(laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DMIL); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX 190); C0
2incubator (FORMA model: 3111); Super-clean bench (safe and sound company of Su Jing group manufactures model: SW-CJ-ZFD); Pure water instrument (Sprlng 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 (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) YAC-1 cell uses DMEM+10%FBS in 37 DEG C, 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 DEG C 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 DEG C, 5%C0
2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add treating piles with 9-medicinal herbs sheet 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, pats dry gently with absorbent paper, and low-speed oscillation 10min on shaking table, as entered 200 l dimethyl sulfoxide, is put in every hole, 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), 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 inspection 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, in the time that dosage reaches 5mg/ml, to YAC-1 cell inhibitory effect variant (P<0.05), dosage this difference in the time of 10mg/ml has significance (P<0.01), has utmost point significant difference (P<0.001) in the time that dosage reaches 15-20mg/ml.
Table 1 treating piles with 9-medicinal herbs sheet is on YAC-1 cell inhibitory effect impact research (X ± SD)
| Group | Drug level (mg/ml) | Suppression ratio (%) |
| Matched group | 0 | 0 |
| 1 | 5 | 8.72±5.47 |
| 2 | 10 | 15.45±9.77* |
| 3 | 15 | 30.41±12.63** |
| 4 | 20 | 41.65±13.25** |
Note: with matched group comparison, * P<O.01; * P<0.001.
5. experiment conclusion
Treating piles with 9-medicinal herbs sheet of the present invention can suppress YAC-1 cell proliferation, reduces the Growth of Cells number of YAC-1 cell, and this effect is dose dependent.
Claims (9)
1. the preparation method of a treating piles with 9-medicinal herbs sheet, made as crude drug by Herba Rubi parvifolii 400g, Rhizoma Polygoni Cuspidati 150g, Radix Sanguisorbae 200g, Liu Jisheng 100g, common fig leaf juice 100g, Flos Chrysanthemi 50g, Ovum Gallus domesticus album 20g, Rhizoma Coptidis 150g, Radix Et Rhizoma Rhei 50g, it is characterized in that, described preparation method is made up of the following step: get Herba Rubi parvifolii, Rhizoma Polygoni Cuspidati, Liu Jisheng, 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 l-3ml/g crude drug min, extraction time 180-220min, obtains supercritical extract, for subsequent use; Get all the other Chinese medicines, pulverize, add 60% ethanol of 5L, 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, 60% ethanol elution, collects 5 times of amount column volume eluents, decompression recycling ethanol, concentrate and be dried, obtain microwave extraction thing, for subsequent use; Above-mentioned supercritical extract and microwave extraction thing are mixed, add starch, 70% ethanol granule processed, dry, tabletting, makes 500, every heavy 0.3g.
2. according to the preparation method of the treating piles with 9-medicinal herbs sheet 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 treating piles with 9-medicinal herbs sheet 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 treating piles with 9-medicinal herbs sheet described in claim l, it is characterized in that described CO
2in supercritical extraction, extraction temperature is 40 DEG C.
5. according to the preparation method of the treating piles with 9-medicinal herbs sheet 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 treating piles with 9-medicinal herbs sheet 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 treating piles with 9-medicinal herbs sheet described in claim l, it is characterized in that, described microwave extracting power is 700W.
8. according to the preparation method of the treating piles with 9-medicinal herbs sheet described in claim l, it is characterized in that, the each extraction time of described microwave extracting is 8 minutes.
9. the treating piles with 9-medicinal herbs sheet described in claim l suppresses the application in mouse lymphoma cell YAC-1 cell proliferation medicine in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410371536.4A CN104116810B (en) | 2014-07-31 | 2014-07-31 | A kind of preparation method for the treatment of piles with 9-medicinal herbs sheet and the application in suppression lymphoma cell YAC-1 cell proliferation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410371536.4A CN104116810B (en) | 2014-07-31 | 2014-07-31 | A kind of preparation method for the treatment of piles with 9-medicinal herbs sheet and the application in suppression lymphoma cell YAC-1 cell proliferation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104116810A true CN104116810A (en) | 2014-10-29 |
| CN104116810B CN104116810B (en) | 2016-03-23 |
Family
ID=51762590
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410371536.4A Active CN104116810B (en) | 2014-07-31 | 2014-07-31 | A kind of preparation method for the treatment of piles with 9-medicinal herbs sheet and the application in suppression lymphoma cell YAC-1 cell proliferation thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104116810B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104306828A (en) * | 2014-11-13 | 2015-01-28 | 山东中大药业有限公司 | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of lymphoma cell YAC-1 cell |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1413710A (en) * | 2002-11-19 | 2003-04-30 | 贵州万胜药业有限责任公司 | Capsule for treating piles with 9-medicinal herbs |
| CN102552478A (en) * | 2010-12-16 | 2012-07-11 | 贵州万胜药业有限责任公司 | Quality detection method of Nine Ingredient Hemorrhoid Capsules |
| CN102813825A (en) * | 2012-08-18 | 2012-12-12 | 王春鹏 | Nine-medicine hemorrhoid pill |
| CN104161852A (en) * | 2014-07-31 | 2014-11-26 | 济南爱思医药科技有限公司 | Preparation method for nine-composition hemorrhoid tablet and application thereof to medicines inhibiting cell proliferation of prostate cancer cell RM-1 |
-
2014
- 2014-07-31 CN CN201410371536.4A patent/CN104116810B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1413710A (en) * | 2002-11-19 | 2003-04-30 | 贵州万胜药业有限责任公司 | Capsule for treating piles with 9-medicinal herbs |
| CN102552478A (en) * | 2010-12-16 | 2012-07-11 | 贵州万胜药业有限责任公司 | Quality detection method of Nine Ingredient Hemorrhoid Capsules |
| CN102813825A (en) * | 2012-08-18 | 2012-12-12 | 王春鹏 | Nine-medicine hemorrhoid pill |
| CN104161852A (en) * | 2014-07-31 | 2014-11-26 | 济南爱思医药科技有限公司 | Preparation method for nine-composition hemorrhoid tablet and application thereof to medicines inhibiting cell proliferation of prostate cancer cell RM-1 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104306828A (en) * | 2014-11-13 | 2015-01-28 | 山东中大药业有限公司 | Preparation method of liver benefiting tablet containing nepal dock root and day lily and application of liver benefiting tablet to preparation of drug for restraining multiplication of lymphoma cell YAC-1 cell |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104116810B (en) | 2016-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102988612B (en) | Preparation method and application of tablet for clearing away heat | |
| CN104116810B (en) | A kind of preparation method for the treatment of piles with 9-medicinal herbs sheet and the application in suppression lymphoma cell YAC-1 cell proliferation thereof | |
| CN104161852A (en) | Preparation method for nine-composition hemorrhoid tablet and application thereof to medicines inhibiting cell proliferation of prostate cancer cell RM-1 | |
| CN103800499A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting melanoma cell B16 from cell proliferation | |
| CN103768211A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting lymphomata cell EL4 from cell proliferation | |
| CN103751424A (en) | Preparation method of cold tablet containing Artemisia carvifolia and holly root, and application of cold tablet in teratoma cell F9 cell proliferation inhibition medicines. | |
| CN103768263A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting cerebral nerve oncocyte Neuro-2a from cell proliferation | |
| CN103893411A (en) | Preparation method and application of green plum cold tablets | |
| CN103768262A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting myeloma cell FO from cell proliferation | |
| CN103800498A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting mouse prostate cancer cell RM-1 from cell proliferation | |
| CN103800816A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting stomach cancer cell MFC from cell proliferation | |
| CN103768266A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting mouse leukaemia cell L6565 from cell proliferation | |
| CN104288658A (en) | Preparation method of fomes officinalis cough and asthma treating tablets and application of fomes officinalis cough and asthma treating tablets in preparation of medicine for inhibiting cell proliferation of mouse myeloma cell SP2/0 | |
| CN103768264A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting teratoma cell P19 from cell proliferation | |
| CN103705825A (en) | Application of lespedeza cuneata turbidity removing tablet in preparation of medicine for inhibiting cell proliferation of embryonal carcinoma F9 | |
| CN103800569A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting lymphoma cell YAC-1 from cell proliferation | |
| CN103751425A (en) | Preparation method of Qingmei cold-treatment tablet and use of Qingmei cold-treatment tablet in drug for inhibiting breast tumor cell C127 proliferation | |
| CN103800770A (en) | Application of boenninghausenia cough tablet in preparing medicine for inhibiting cell proliferation of breast tumor cell C127 | |
| CN103768212A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting stomach cancer cell MFC from cell proliferation | |
| CN103768207A (en) | Preparation method of Qingmei cold tablet and application of Qingmei cold tablet in drugs for inhibiting lung cancer cell Hepa1-6 from cell proliferation | |
| CN103768514A (en) | Preparation method of Danshitongli tablet and application of Danshitongli tablet in preparing drugs for inhibiting myeloma cell SP2/0 from cell proliferation | |
| CN104352931A (en) | Preparation method of fomitopsis officinalis tablet capable of treating cough and asthma and application thereof in preparing drug for inhibiting cell proliferation of mouse melanoma cell B16 | |
| CN104288659A (en) | Preparation method of fomes officinalis cough and asthma treating tablets and application of fomes officinalis cough and asthma treating tablets in preparation of medicine for inhibiting cell proliferation of mouse breast tumor cell C127 | |
| CN104352928A (en) | Preparation method of fomes officinalis cough and asthma tablet and application of fomes officinalis cough and asthma tablet in inhibiting mouse forestomach carcinoma cell proliferation drug | |
| CN104288653A (en) | Preparation method of fomes officinalis cough and asthma treating tablets and application of fomes officinalis cough and asthma treating tablets in preparation of medicine for inhibiting cell proliferation of mouse myeloma cell FO |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240102 Address after: Room 2102, block a, building 1, Jinan pharmaceutical Valley R & D platform area, No.1, north section of Gangxing Third Road, comprehensive bonded zone, Jinan high tech Zone, Shandong Province Patentee after: Jinan Xinglu Medical Technology Co.,Ltd. Address before: Room B402, Comprehensive Building, University Science and Technology Park, No. 750 Shunhua Road, High tech Zone, Jinan City, Shandong Province, 250101 Patentee before: JINAN ASIA PHARMA TECH Co.,Ltd. |