CN100352458C - Preparaton of banlangen and method for controlling quality - Google Patents
Preparaton of banlangen and method for controlling quality Download PDFInfo
- Publication number
- CN100352458C CN100352458C CNB031356885A CN03135688A CN100352458C CN 100352458 C CN100352458 C CN 100352458C CN B031356885 A CNB031356885 A CN B031356885A CN 03135688 A CN03135688 A CN 03135688A CN 100352458 C CN100352458 C CN 100352458C
- Authority
- CN
- China
- Prior art keywords
- radix isatidis
- preparation
- adenosine
- eye drop
- quality control
- 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.)
- Expired - Lifetime
Links
- 239000010231 banlangen Substances 0.000 title claims abstract description 113
- 238000000034 method Methods 0.000 title claims description 33
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims abstract description 122
- 238000002360 preparation method Methods 0.000 claims abstract description 75
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims abstract description 61
- 229960005305 adenosine Drugs 0.000 claims abstract description 61
- 238000003908 quality control method Methods 0.000 claims abstract description 15
- 239000003889 eye drop Substances 0.000 claims description 46
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Substances OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 29
- 238000012360 testing method Methods 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- 239000007788 liquid Substances 0.000 claims description 23
- 239000000243 solution Substances 0.000 claims description 22
- 239000008187 granular material Substances 0.000 claims description 15
- 238000002347 injection Methods 0.000 claims description 12
- 239000007924 injection Substances 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 10
- 239000011550 stock solution Substances 0.000 claims description 10
- 239000012071 phase Substances 0.000 claims description 9
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 8
- 239000002775 capsule Substances 0.000 claims description 8
- 239000000499 gel Substances 0.000 claims description 7
- 239000002904 solvent Substances 0.000 claims description 7
- 238000005303 weighing Methods 0.000 claims description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 239000013558 reference substance Substances 0.000 claims description 6
- 239000006188 syrup Substances 0.000 claims description 6
- 235000020357 syrup Nutrition 0.000 claims description 6
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 229910021529 ammonia Inorganic materials 0.000 claims description 4
- 239000006187 pill Substances 0.000 claims description 3
- 239000003826 tablet Substances 0.000 claims description 3
- 229910019142 PO4 Inorganic materials 0.000 claims description 2
- 238000004587 chromatography analysis Methods 0.000 claims description 2
- 238000010812 external standard method Methods 0.000 claims description 2
- 239000007791 liquid phase Substances 0.000 claims description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 2
- 239000010452 phosphate Substances 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 41
- 230000000694 effects Effects 0.000 abstract description 19
- 239000000284 extract Substances 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 abstract description 5
- 239000004615 ingredient Substances 0.000 abstract description 5
- 230000003285 pharmacodynamic effect Effects 0.000 abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 41
- 239000012567 medical material Substances 0.000 description 33
- 229940079593 drug Drugs 0.000 description 29
- CRDNMYFJWFXOCH-YPKPFQOOSA-N (3z)-3-(3-oxo-1h-indol-2-ylidene)-1h-indol-2-one Chemical compound N/1C2=CC=CC=C2C(=O)C\1=C1/C2=CC=CC=C2NC1=O CRDNMYFJWFXOCH-YPKPFQOOSA-N 0.000 description 18
- 239000000706 filtrate Substances 0.000 description 17
- 241000894006 Bacteria Species 0.000 description 15
- 230000000844 anti-bacterial effect Effects 0.000 description 15
- 238000000605 extraction Methods 0.000 description 15
- 239000001963 growth medium Substances 0.000 description 11
- 238000004321 preservation Methods 0.000 description 11
- 239000000523 sample Substances 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 10
- 239000000047 product Substances 0.000 description 10
- 229920001817 Agar Polymers 0.000 description 9
- CRDNMYFJWFXOCH-BUHFOSPRSA-N Couroupitine B Natural products N\1C2=CC=CC=C2C(=O)C/1=C1/C2=CC=CC=C2NC1=O CRDNMYFJWFXOCH-BUHFOSPRSA-N 0.000 description 9
- 239000008272 agar Substances 0.000 description 9
- 238000003556 assay Methods 0.000 description 9
- CRDNMYFJWFXOCH-UHFFFAOYSA-N isoindigotin Natural products N1C2=CC=CC=C2C(=O)C1=C1C2=CC=CC=C2NC1=O CRDNMYFJWFXOCH-UHFFFAOYSA-N 0.000 description 9
- 235000000177 Indigofera tinctoria Nutrition 0.000 description 8
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 8
- 229940097275 indigo Drugs 0.000 description 8
- COHYTHOBJLSHDF-UHFFFAOYSA-N indigo powder Natural products N1C2=CC=CC=C2C(=O)C1=C1C(=O)C2=CC=CC=C2N1 COHYTHOBJLSHDF-UHFFFAOYSA-N 0.000 description 8
- 241000588724 Escherichia coli Species 0.000 description 7
- 238000010790 dilution Methods 0.000 description 7
- 239000012895 dilution Substances 0.000 description 7
- 235000010603 pastilles Nutrition 0.000 description 7
- 241000588747 Klebsiella pneumoniae Species 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 241000191967 Staphylococcus aureus Species 0.000 description 6
- 241000700605 Viruses Species 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 231100000915 pathological change Toxicity 0.000 description 6
- 230000036285 pathological change Effects 0.000 description 6
- 229960004150 aciclovir Drugs 0.000 description 5
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 description 5
- 239000002552 dosage form Substances 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 239000010985 leather Substances 0.000 description 5
- 238000004064 recycling Methods 0.000 description 5
- 238000013112 stability test Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 241000700588 Human alphaherpesvirus 1 Species 0.000 description 4
- 230000001986 anti-endotoxic effect Effects 0.000 description 4
- 210000001508 eye Anatomy 0.000 description 4
- 229940012356 eye drops Drugs 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 241000588624 Acinetobacter calcoaceticus Species 0.000 description 3
- 241000334154 Isatis tinctoria Species 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000001568 sexual effect Effects 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000588697 Enterobacter cloacae Species 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 241000334160 Isatis Species 0.000 description 2
- 201000007100 Pharyngitis Diseases 0.000 description 2
- 241000700584 Simplexvirus Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000191963 Staphylococcus epidermidis Species 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 241000257673 Strobilanthes cusia Species 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 239000008186 active pharmaceutical agent Substances 0.000 description 2
- 102000011759 adducin Human genes 0.000 description 2
- 108010076723 adducin Proteins 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 230000001174 ascending effect Effects 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 235000008504 concentrate Nutrition 0.000 description 2
- 239000012531 culture fluid Substances 0.000 description 2
- 230000000249 desinfective effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000857 drug effect Effects 0.000 description 2
- 229960001617 ethyl hydroxybenzoate Drugs 0.000 description 2
- NUVBSKCKDOMJSU-UHFFFAOYSA-N ethylparaben Chemical compound CCOC(=O)C1=CC=C(O)C=C1 NUVBSKCKDOMJSU-UHFFFAOYSA-N 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 230000001408 fungistatic effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- GYCKQBWUSACYIF-UHFFFAOYSA-N o-hydroxybenzoic acid ethyl ester Natural products CCOC(=O)C1=CC=CC=C1O GYCKQBWUSACYIF-UHFFFAOYSA-N 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 241000701161 unidentified adenovirus Species 0.000 description 2
- KZJWDPNRJALLNS-VPUBHVLGSA-N (-)-beta-Sitosterol Natural products O[C@@H]1CC=2[C@@](C)([C@@H]3[C@H]([C@H]4[C@@](C)([C@H]([C@H](CC[C@@H](C(C)C)CC)C)CC4)CC3)CC=2)CC1 KZJWDPNRJALLNS-VPUBHVLGSA-N 0.000 description 1
- CSVWWLUMXNHWSU-UHFFFAOYSA-N (22E)-(24xi)-24-ethyl-5alpha-cholest-22-en-3beta-ol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)C=CC(CC)C(C)C)C1(C)CC2 CSVWWLUMXNHWSU-UHFFFAOYSA-N 0.000 description 1
- KLEXDBGYSOIREE-UHFFFAOYSA-N 24xi-n-propylcholesterol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CCC)C(C)C)C1(C)CC2 KLEXDBGYSOIREE-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 208000006400 Arbovirus Encephalitis Diseases 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- LPZCCMIISIBREI-MTFRKTCUSA-N Citrostadienol Natural products CC=C(CC[C@@H](C)[C@H]1CC[C@H]2C3=CC[C@H]4[C@H](C)[C@@H](O)CC[C@]4(C)[C@H]3CC[C@]12C)C(C)C LPZCCMIISIBREI-MTFRKTCUSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- ARVGMISWLZPBCH-UHFFFAOYSA-N Dehydro-beta-sitosterol Natural products C1C(O)CCC2(C)C(CCC3(C(C(C)CCC(CC)C(C)C)CCC33)C)C3=CC=C21 ARVGMISWLZPBCH-UHFFFAOYSA-N 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 206010052369 Encephalitis lethargica Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241001360526 Escherichia coli ATCC 25922 Species 0.000 description 1
- 208000005100 Herpetic Keratitis Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 208000005647 Mumps Diseases 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 206010073938 Ophthalmic herpes simplex Diseases 0.000 description 1
- 241000589614 Pseudomonas stutzeri Species 0.000 description 1
- 229920002125 Sokalan® Polymers 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 208000016150 acute pharyngitis Diseases 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000003288 anthiarrhythmic effect Effects 0.000 description 1
- 230000003460 anti-nuclear Effects 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- MJVXAPPOFPTTCA-UHFFFAOYSA-N beta-Sistosterol Natural products CCC(CCC(C)C1CCC2C3CC=C4C(C)C(O)CCC4(C)C3CCC12C)C(C)C MJVXAPPOFPTTCA-UHFFFAOYSA-N 0.000 description 1
- LGJMUZUPVCAVPU-UHFFFAOYSA-N beta-Sitostanol Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(C)CCC(CC)C(C)C)C1(C)CC2 LGJMUZUPVCAVPU-UHFFFAOYSA-N 0.000 description 1
- NJKOMDUNNDKEAI-UHFFFAOYSA-N beta-sitosterol Natural products CCC(CCC(C)C1CCC2(C)C3CC=C4CC(O)CCC4C3CCC12C)C(C)C NJKOMDUNNDKEAI-UHFFFAOYSA-N 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 229960001631 carbomer Drugs 0.000 description 1
- 108010079058 casein hydrolysate Proteins 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229940126678 chinese medicines Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 210000004349 growth plate Anatomy 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 208000010805 mumps infectious disease Diseases 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 238000005325 percolation Methods 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000007560 sedimentation technique Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 description 1
- 229950005143 sitosterol Drugs 0.000 description 1
- NLQLSVXGSXCXFE-UHFFFAOYSA-N sitosterol Natural products CC=C(/CCC(C)C1CC2C3=CCC4C(C)C(O)CCC4(C)C3CCC2(C)C1)C(C)C NLQLSVXGSXCXFE-UHFFFAOYSA-N 0.000 description 1
- 235000015500 sitosterol Nutrition 0.000 description 1
- 210000002460 smooth muscle Anatomy 0.000 description 1
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 1
- 235000010234 sodium benzoate Nutrition 0.000 description 1
- 239000004299 sodium benzoate Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 238000009827 uniform distribution Methods 0.000 description 1
- 201000002498 viral encephalitis Diseases 0.000 description 1
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention provides a radix isatidis preparation which is prepared from traditional Chinese medicine of radix isatidis or a radix isatidis extract, wherein in every preparation unit, the content of adenosine is from 0.075 to 2 mg, and the content of adenosine in the radix isatidis is at least 0.3 mg/g. The adenosine is adopted as an index ingredient, so the reproducibility is high, and the stability can be controlled. The experiment of the pharmacodynamics proves that the adenosine is not the effective component, but the quality and the curative effect of the radix isatidis preparation can be controlled from the lateral surface by controlling the content of the adenosine; thus, the preparation is more steady and controllable. The present invention provides a new quality control method for radix isatidis and the radix isatidis preparation.
Description
Technical field
The invention provides Radix Isatidis preparation and method of quality control thereof, belong to the field of Chinese medicines.
Background technology
Radix Isatidis is the dry root of cruciferae isatis (Isatis indigotica Fort.), and bitter in the mouth is cold in nature, goes into the heart, stomach warp, has heat-clearing and toxic substances removing, removing heat from blood repercussive effect.Main chemical compositions has indigo glycosides, indirubin, sitosterol, several amino acids, adenosine, polysaccharide etc.Be widely used in treatment influenza, mumps, acute and chronic pharyngitis, herpes simplex keratitis, epidemic encephalitis B inflammation etc. clinically.Pharmacologically active has antiendotoxin, antiviral, antitumor, immunomodulating etc.Radix Isatidis preparation has Radix Isatidis injection, Radix Isatidis granule, Radix Isatidis capsule, indigowoad root oral liquid, Radix Isatidis syrup, Radix Isatidis tablet, composite capsule of radix isatidis, compound isatis root granules, Compound recipe Radix Isatidis sheet etc.
Radix Isatidis preparation adopts the decoction and alcohol sedimentation technique preparation more at present, adopts percolation on a small quantity.Radix Isatidis preparation adopts the content of aminoacid or indigo, indirubin as quality index.Aminoacid is a kind of popular composition, does not have specificity as quality index, and the specificity difference is especially in compound preparation; And adopt indigo or indirubin as quality index, because indigo and indirubin is fat-soluble composition, adopt decocting in water to be difficult for stripping, extraction ratio is low, detected at all does not go out indirubin (Zhang Runzhen in some preparation, Zhang Yuwen. the progress of plate grass roots. Chinese herbal medicine, 2000,31 (6): 474-476), what have more that bibliographical information says Isatis indigotica Fort. does not have an indirubin and indigo at all, in its preparation detected indirubin and indigo amount be its gather and the course of processing in sneak into indirubin contained in rhizome and its incidental petiole residual base and indigo amount, and both do not have antiendotoxin activity (Zhang Hanming, Zhang Ge etc. Radix Isatidis and Folium Isatidis different parts indigo, the anti-endotoxin effect of indirubin assay and part composition thereof is poor. the pharmacy practice magazine, 2000,18 (5): 347).
Adenosine has another name called ribosidoadenine, is a kind of known natural component, molecular formula C
10H
13N
5O
4, molecular weight 267.24, white crystals, little have a salty bitterness, soluble in water, is dissolved in ethanol and ether hardly, and melting point 234-235 ℃, specific optical rotation-61.7 (C=0.706% is in the water).Adenosine is except that having anticoagulation, coronary artery dilator, effects such as lax bronchial smooth muscle, calm nervus centralis and anti-arrhythmia, confirm again to stop effectively by the inductive leukocyte of PAF-endotheliocyte adhere to and participate in antiinflammation (Xu Ruming, Lu Yang etc. the content of adenosine in the determined by ultraviolet spectrophotometry Bulbus Fritillariae Uninbracteatae, CHINA JOURNAL OF CHINESE MATERIA MEDICA 1996,21 (6): 556).
Safely, effectively, stable, controlled is the standard of modern medicines preparation, at present the Radix Isatidis preparation report has the effect of aspects such as antiviral, antibacterial, antiendotoxin, and it possesses safe and effective characteristics clinical proof, but because contained composition complexity in the Radix Isatidis, make Radix Isatidis medical material and preparation stability thereof poor, different medical materials, finished product batch difference, quality also have bigger floating, and make the poor controllability of Radix Isatidis preparation.
Summary of the invention
In order to address the above problem, technical scheme to be solved by this invention provides Radix Isatidis preparation and method of quality control thereof.
The invention provides Radix Isatidis preparation, it is to be prepared from by Chinese medicine Radix Isatidis Radix Isatidis medical material or extract, and the content range of adenosine is 0.075～2mg in wherein every preparation unit, the content 〉=0.3mg/g of adenosine in the Radix Isatidis medical material.
Wherein said preparation unit is meant every ml, the every ml in the eye drop among in every gram among in every ml in the oral liquid, the capsule every, granule, the tablet every, syrup.
Described preparation is eye drop, original position gel eye drop, injection, granule, capsule, oral liquid, syrup, tablet, pill, powder.Further, described preparation is an eye drop, and the content range of adenosine is 0.0998～2mg in the described eye drop.
The present invention also provides the method for quality control of Radix Isatidis preparation, and it is that the employing adenosine is that the index composition carries out qualitative and quantitative control, and the content range of adenosine is 0.075～2mg in every preparation unit.
The method of described qualitative control is: adopting chloroform, ethyl acetate, isopropyl alcohol, water, ammonia is developing solvent, uses silica gel G F
254Lamellae (self-control) carries out one dimension to launch, and reference standards is an adenosine, and under uviol lamp, wavelength is that 254nm inspects, and in the test sample chromatograph, is showing identical skin dark stain with contrast chromatograph corresponding position.Further, this qualitative control method is: it is an amount of to get Radix Isatidis preparation, and thin up is 10ml, the full n-butanol extraction that closes of water three times, be respectively 20ml, 20ml, 10ml, separatory merges n-butanol layer, water bath method, residue 70% dissolve with ethanol is as need testing solution; Other gets adenosine reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 878-20000), add 70% ethanol and make the solution that every ml contains 1mg, in contrast product solution; According to the test of 2000 editions one appendix VIB thin layer chromatography of Chinese Pharmacopoeia, draw each 10 μ l of above-mentioned two kinds of solution, respectively o'clock in a same silica gel G F
254On the lamellae, with chloroform-ethyl acetate-isopropyl alcohol-water-ammonia water, volume ratio is 8: 2: 6: 0.5: 0.12, as developing solvent, ascending development takes out, and dries, under uviol lamp, wavelength is that 254nm inspects, and in the test sample chromatograph, is showing identical skin dark stain with contrast chromatograph corresponding position.
The method of described quantitative control is:
Chromatographic condition:
Instrument: HP1000 high performance liquid chromatograph, G1310A IsoPump pump, G1314A VMD UV-detector; The Chemstation data processing software;
Chromatographic column: Alltima C18 5 μ m, 150mm * 4.6mm;
Mobile phase: methanol-water, volume ratio are 5～12: 88～95 or phosphate buffer-methanol, and volume ratio is 15～20: 3～10; Flow velocity: 1ml/min;
Column temperature: room temperature;
Detect wavelength: 260nm;
Sample size: 20 μ l;
The preparation of need testing solution: the accurate Radix Isatidis preparation of drawing, put in the 10ml volumetric flask, be diluted to scale with 30% methanol, shake up, promptly;
The preparation of reference substance solution: precision takes by weighing the about 10mg of the adenosine that is dried to constant weight, puts in the 50ml volumetric flask, adds mobile phase and dissolves surely and be diluted to scale, shakes up, in contrast the product stock solution;
Get above solution and inject the efficient liquid phase chromatographic analysis instrument, measure, calculate by external standard method, promptly.
Further, described mobile phase: methanol-water, volume ratio are 8: 92.
It is the index sexual element that Radix Isatidis preparation of the present invention adopts adenosine, measures by the qualitative, quantitative to adenosine, can reflect Radix Isatidis medical material and stability of formulation thereof, reaches the purpose of control of quality, repeatability height, stable and controllable; Prove by pharmacodynamics test, though adenosine is not an effective ingredient,, can control the curative effect of Radix Isatidis preparation from the side by control to adenosine content, make preparation more stable, controlled, for Radix Isatidis medical material and preparation thereof provide a kind of new method of quality control.
Obviously, according to foregoing of the present invention,,, can also make modification, replacement or the change of other various ways not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite according to the ordinary skill knowledge and the customary means of this area.
The specific embodiment of form is described in further detail foregoing of the present invention again by the following examples.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.
Description of drawings
The qualitative control thin-layer chromatogram of Fig. 1 Radix Isatidis eye drop: point 1 is the adenosine standard substance, and point 2 is not for containing the negative control of adenosine, and point 3,4,5 is a Radix Isatidis eye drop sample.
The specific embodiment
The preparation of experimental example 1 Radix Isatidis primary crude drug powder
Radix Isatidis medical material cold drying (below 60 ℃), powder is beaten in stripping and slicing, crosses sieve No. 2, promptly.
The preparation of experimental example 2 Radix Isatidis eye drops
Get Radix Isatidis 500g, add 10 times of water gagings and decoct twice, 2 hours for the first time, 1 hour for the second time, collecting decoction filtered, filtrate is concentrated into about 750ml, and cooling adds ethanol and makes and contain alcohol amount and reach 70%, stir evenly, cold preservation 24 hours filters, filtrate recycling ethanol to 350～500ml, cold preservation is spent the night, and filters, it is 8.0～8.5 that filtrate is regulated pH value with ammonia solution, stirs evenly cold preservation 48 hours, filter, add heat extraction ammonia, add the 1g ethyl hydroxybenzoate, adding distil water is to 1000ml, sterilization, filter with G4 sintered filter funnel, 0.22um filter membrane, embedding, promptly.
The preparation of experimental example 3 Radix Isatidis original position gel eye drops
Get Radix Isatidis 500g, add 10 times of water gagings and decoct twice, 2 hours for the first time, 1 hour for the second time, collecting decoction, filter, filtrate is concentrated into about 750ml, and cooling adds ethanol and makes and contain alcohol and measure and reach 70%, stirs evenly, cold preservation 24 hours filters, filtrate recycling ethanol to 350～500ml, and cold preservation is spent the night, and filters; Other gets carbomer 3g and adds the abundant swelling of an amount of distilled water, hydroxypropyl methylcellulose 12g and the dissolving of ethyl hydroxybenzoate 1g adding distil water, merge above-mentioned solution, add sodium chloride 8.5g and regulate osmotic pressure, adding distil water is to 1000ml, and it is 5.5-6.0 that sodium hydroxide is regulated pH, sterilization, filter with G4 sintered filter funnel, 0.22um filter membrane, embedding, promptly.
So-called original position gel eye drop is meant: the pH value of most of eye drops is identical with the pH value of people's tear usually, meta-alkalescence, and pH value is between 7.3～7.5, if pH value is improper, can cause stimulation to eye, increase the secretion of tear, cause medicine to be rinsed loss, even the damage cornea.PH value of the present invention is 5.5-6.0, slant acidity, be because the present invention adopts original position gel technology to prepare eye drop, make this pharmaceutical dosage form under acid condition, be the solution form, under the distinctive physiological environment effect of eye, influenced by the pH of tear behind the uniform distribution during medicine liquid droplet is pleasing to the eye, form gel, be covered in eyeball surface, and do not influence the normal vision of eyes, drug slow discharges, and reaches the effect of long-acting slow-release and can reduce in the use pollution to preparation itself.
The preparation of experimental example 4 Radix Isatidis tablets
Get Radix Isatidis medical material 1000g, decoct with water 2 times, 1 hour for the first time, 45 minutes for the second time, filter, merging filtrate is concentrated into 1000ml.Be placed to room temperature, add 95% ethanol and make and contain alcohol amount and reach 60%.Cold preservation filters, and filtrate recycling ethanol is concentrated into the thick paste shape, adds starch 200g and other right amount of auxiliary materials, makes granule, suppresses 1000 in blocks, coating, promptly.
The preparation of experimental example 5 Radix Isatidis granules
Get Radix Isatidis medical material 500g, decoct with water 2 times (2 hours for the first time, 1 hour for the second time), filter, merging filtrate suitably concentrates.Add ethanol and make and contain alcohol amount and reach 60%, get supernatant and reclaim ethanol, being concentrated into relative density is the thick paste of 1.33 (80 ℃), adds adjuvant (1 part of clear paste, 2 parts of sucrose, 1.3 parts in dextrin) and granulates, promptly.
The preparation of experimental example 6 Radix Isatidis syrup
Get Radix Isatidis medical material 700g, decoct with water 2 times (2 hours for the first time, 1 hour for the second time), filter, merging filtrate leaves standstill.Get supernatant and suitably concentrate, add correctives (sucrose 400g), antiseptic (sodium benzoate 3g), the dissolving back filters, and adds water and is adjusted to prescribed volume (1000ml), promptly.
The preparation of experimental example 7 Radix Isatidis injections
Get Radix Isatidis medical material 500g, decoct with water 2 times, 2 hours for the first time, 1 hour for the second time, merge decoction liquor, filter, filtrate is concentrated into about 750ml, and cooling adds ethanol and makes the alcohol amount 70% that contains, stir evenly, left standstill 24 hours, filter, filtrate recycling ethanol to 350～500ml, cold preservation filters, it is 8.0～8.5 that filtrate is regulated pH value with ammonia solution, stirs evenly cold preservation 48 hours, add heat extraction ammonia, add the injection water to 900ml, cold preservation filters, it is 7.0～7.5 that filtrate is regulated pH value with 1% sodium hydroxide solution, and cold preservation filters, filtrate is with mannitol 10g, and adds the injection water and adjust total amount to 1000ml, filters, embedding, sterilization, promptly.
The preparation of experimental example 8 Radix Isatidis capsules
Get Radix Isatidis medical material 1000g, decoct with water 2 times, 1 hour for the first time, 45 minutes for the second time, filter, merging filtrate is concentrated into 1000ml.Be placed to room temperature, add 95% ethanol and make and contain alcohol amount and reach 60%.Cold preservation filters, and filtrate recycling ethanol is concentrated into the thick paste shape, adds starch 200g and other right amount of auxiliary materials, makes granule, is filled in capsule promptly.
The qualitative control of adenosine in experimental example 9 Radix Isatidis preparations
Treating excess syndrome is tested the 2 eye drop 5ml that make, and thin up is 10ml, and three times (10ml), separatory merges n-butanol layer to the full n-butanol extraction that closes of water for 20ml, 20ml, and water bath method, residue 70% dissolve with ethanol is as need testing solution.Other gets the adenosine reference substance, adds 70% ethanol and makes the solution that every ml contains 1mg, in contrast product solution.According to thin layer chromatography (2000 editions appendix VIB of Chinese Pharmacopoeia) test, draw each 10 μ l of above-mentioned two kinds of solution, respectively o'clock in a same silica gel G F
254On the lamellae, be developing solvent with chloroform-ethyl acetate-isopropyl alcohol-water-ammonia water (8: 2: 6: 0.5: 0.12), ascending development takes out, and dries, and inspects under uviol lamp (254nm).In the test sample chromatograph, showing identical skin dark stain with contrast chromatograph corresponding position.(see figure 1)
The assay of adenosine in the experimental example 10 Radix Isatidis eye drops
1. chromatographic condition: instrument: HP1100 high performance chromatograph, G1310A IsoPump pump, G1314A
The VMD UV-detector.The Chemstation data processing software.
Chromatographic column: Alltima C18 (5 μ m, 150mm * 4.6mm); Mobile phase: methanol-water (8: 92); Flow velocity: 1ml/min; Column temperature: room temperature.Detect wavelength: 260nm.
2. the preparation of need testing solution: the eye drop that accurate absorption experimental example 2 makes is made sample 2ml, puts in the 10ml volumetric flask, is diluted to scale with 30% methanol, shakes up, promptly.
3. the preparation of reference substance solution: precision takes by weighing the about 10mg of the adenosine that is dried to constant weight, puts in the 50ml volumetric flask, adds mobile phase and dissolves surely and be diluted to scale, shakes up, in contrast the product stock solution.
4. the investigation of linear relationship:
y＝60659x-8.5766，r＝0.9998
5. stable:
Sample solution is stable in 8 hours
6. repeatability:
The RSD% of 3 duplicate samples is less than 3%, and the sample repeatability is good.
7. the response rate:
The response rate of sample is between 95%～105%, and test shows that this law has the good response rate.
8. assay:
Three batch sample measurement results:
| Sample number | Content |
| 1 2 3 | 0.1124 0.1157 0.1045 |
According to measurement result, produce actually in conjunction with industry is big, decide the every ml of this product and contain adenosine and must not be less than 0.0998mg.
Adenosine is quantitatively controlled in the experimental example 11 Radix Isatidis medical materials
L, get the Radix Isatidis medical material: this product is the dry root of cruciferae isatis Isatis indigotica Fort..Purchase in Chengdu lotus pond Chinese Medicinal Materials Markets.This product is that pharmacopeia 2000 editions is recorded kind, should meet " pertinent regulations under the 163rd page of Baphicacanthus cusia rootkey of Chinese pharmacopoeia version in 2000.For ensureing the needs of the quality of the pharmaceutical preparations, again the content of its adenosine is stipulated.
2, the assay of adenosine: get 2g medical material+20ml30% methanol, standardize solution is to 25ml
2.1 chromatographic condition: with experimental example 10 described assay conditions.
2.2 extraction choice of Solvent: having investigated with water, 10% methanol, 30% methanol is the extraction ratio that extracts solvent, and the result shows that the extraction efficiency of 30% methanol is the highest, so select 30% methanol as extracting solvent.
2.3 the selection of extracting method: investigated reflux, extract, and supersound extraction, the result shows that the extraction efficiency of two kinds of methods is almost consistent, but supersound extraction is easier, and easy operating is so select supersound extraction.
2.4 the selection of extraction time: investigated supersound extraction respectively 30,45,60 minutes, the result shows, extracts in 45 minutes medical materials adenosine and just extracts substantially and use up, so selective extraction 45 minutes.
2.5 the assay of medical material
By method suggested, 4 batches of medical materials are carried out assay, the results are shown in Table.
Medical material assay result (n=2)
| Sample number | 1 | 2 | 3 | 4 |
| Content (mg/g) | 0.3252 | 0.3806 | 0.3016 | 0.3142 |
| Average content (mg/g) | 0.3304 | |||
By above measurement result, in conjunction with actual, the content of adenosine must not be lower than 0.3mg/g in the Radix Isatidis medical material.By mensuration, can reflect the quality of Radix Isatidis medical material to adenosine content in the Radix Isatidis medical material.
The stability test of adenosine in experimental example 12 Radix Isatidis medical materials and the Radix Isatidis granule
1. getting each 3 batches of Radix Isatidis medical material and Radix Isatidis granules, to place temperature be to carry out accelerated stability test under the condition of 37-40 ℃ and relative humidity 75%, investigated continuous 3 months 1 time in every month.
2. mainly investigation project is an assay, and data see the following form.
3. chromatographic condition is with experimental example 10:
The preparation of test sample solution: precision takes by weighing the granule 1g that experimental example 1 medical material granule 1g and experimental example 5 make, and the 5m30% methanol extraction filters respectively, and filtrate is put in the 10ml volumetric flask, is diluted to scale with 30% methanol, shakes up, promptly.
The preparation of reference substance solution: precision takes by weighing the about 10mg of the adenosine that is dried to constant weight, puts in the 50ml volumetric flask, adds mobile phase and dissolves surely and be diluted to scale, shakes up, in contrast the product stock solution.
Table 1 Radix Isatidis granule accelerated stability test result (mg/g)
| Standing time (moon) | 0 | 1 | 2 | 3 | |
| Batch | 01 02 03 | 0.1024 0.1020 0.1023 | 0.1022 0.1019 0.1024 | 0.1024 0.1020 0.1023 | 0.1021 0.1018 0.1022 |
Table 2 Radix Isatidis medical material accelerated stability test result (mg/g)
| Standing time (moon) | 0 | 1 | 2 | 3 | |
| Batch | 01 02 03 | 0.3252 0.3806 0.3142 | 0.3252 0.3805 0.3141 | 0.3251 0.3804 0.3140 | 0.3252 0.3804 0.3141 |
4. conclusion:
According to the accelerated stability test result as can be known, the method for adenosine content is stablized feasible in above-mentioned detection Baphicacanthus cusia root herb and the preparation thereof.
Below further prove beneficial effect of the present invention by concrete pharmacodynamics test.
The in-vitro antibacterial test of embodiment 1 Radix Isatidis eye drop
One, experiment material
1, trial drug
(1), the Radix Isatidis eye drop: rufous liquid, specification: 8ml/ prop up (containing crude drug 4g/8ml), lot number: 030401, by experimental example 2 methods preparations, and example 10 is measured the wherein content of adenosine by experiment.Draw 20ml during test, after adding 50 ℃ of M-H agar (caseinhydrolysate agar) culture medium mixing of 20ml thawing, sucking-off 20ml pastille culture medium is toppled over plate from 40ml, add 20ml in the remaining 20ml pastille culture medium again and do not have the dilution of medicine agar culture medium, behind the mixing again sucking-off 20ml topple over plate, the rest may be inferred, promptly with this doubling dilution prepare contain Radix Isatidis eye drop dilution factor be stock solution 250,125,62.5,31.3 ... 0.24 serial pastille plate stand-by.
(2), Radix Isatidis injection: rufous liquid, crude drug content are that (every 2ml is equivalent to Radix Isatidis 1g to 500mg crude drug/ml, and lot number 20030416 is produced by Shiyan Kang Di pharmaceutical Co. Ltd of the strong bright Pharmaceutical in Wuhan group.Draw 20ml during test, after adding 50 ℃ of M-H agar culture medium mixings of 20ml thawing, sucking-off 20ml pastille culture medium is toppled over plate from 40ml, add 20ml in the remaining 20ml pastille culture medium again and do not have the dilution of medicine agar culture medium, behind the mixing again sucking-off 20ml topple over plate, the rest may be inferred, promptly with this doubling dilution prepare crude drug content be 250,125,62.5 ... 0.5mg the serial pastille plate of crude drug/ml is stand-by.
2, antibacterial
(1) clinical isolates strain: the test bacterial strain uses therefor is the clinical separation pathogenic bacterium that 2001-2003 collects from the area, Sichuan, all strains are being collected isolating unit (Hospital Affiliated To Chengdu Traditional Chinese Medicine Univ clinical laboratory Bacteriology Room) all after identifying, again identify that with API system (the API system is meant the Bacteria Identification system, is the Vitek automatic bacterial identification systems of being produced by French Mei Liai company) back uses through this chamber again.
Staphylococcus aureus 20 strains, staphylococcus epidermidis 5 strains, escherichia coli 13 strains, 6 strains of ESBLs escherichia coli, Klebsiella Pneumoniae 6 strains, 6 strains of ESBLs Klebsiella Pneumoniae, false unit cell 4 strains of Amur, aerobacteria 5 strains, enterobacter cloacae 7 strains, husky thunder bacterium 2 strains, acinetobacter calcoaceticus 20 strains, totally 94 strains.
(2) standard Quality Control bacterial strain: it is that pharmacological room of Chengdu University of Traditional Chinese Medicine preserves bacterial strain that staphylococcus aureus ATCC25923, escherichia coli ATCC25922, ESBLs Klebsiella Pneumoniae ATCC700603 and standard are produced enzyme strain SHV-1, SHV-2.
3, culture medium
The M-H culture medium is produced by the extensive and profound in meaning star biotechnology in Beijing Co., Ltd.
The M-H broth bouillon: take by weighing 25g, add the 1000ml distilled water, the mixing packing is adjusted pH value to 7.2-7.4, and 121 ℃ of autoclavings 20 minutes are used to remove from office the drug sensitive test of the blue positive, negative aerobe.
The M-H solid medium: take by weighing 36.5g, add the 1000ml distilled water, the mixing packing is adjusted pH value to 7.2-7.4, and 121 ℃ of autoclavings 20 minutes are used to remove from office the drug sensitive test of the blue positive, negative aerobe.
Two, test method
1, the mensuration of minimal inhibitory concentration
Adopt the agar doubling dilution (see " microbiology " 128 pages, Shanghai science tech publishing house, in May, 1994) measure the minimum inhibitory concentration (MIC) of Radix Isatidis eye drop.In the agar plate surface that contains different pharmaceutical concentration, every some bacteria containing amount is about 10 with microbionation
5(CFU is colony-forming units to CFU/ml, is the conventional unit of colony counting method.), hatch 18-24 hour observed result for 37 ℃, be the minimum inhibitory concentration (MIC value) of medicine with the least concentration of contained drug in the no bacterial growth plate culture medium to this bacterium.
2, the mensuration of minimum bactericidal concentration (MBC)
Adopt liquid diluting method (seeing " microbiology " 129 pages, Shanghai science tech publishing house, in May, 1994) to measure the minimal bactericidal concentration (MBC) of Radix Isatidis eye drop.After will being subjected to reagent liquid to carry out doubling dilution with M-H meat soup, (making final concentration is about 10 to be tried bacterium liquid respectively at inoculation 100ul in the 2ml pastille culture fluid
5CFU/ml), place 37 ℃ to hatch 18-20 hour, perusal is the minimum inhibitory concentration (MIC value) of medicine to this bacterium with the least concentration of contained drug in the no bacterial growth test tube.
The 0.01ml culture fluid that perusal is not had the test tube of bacterial growth is applied to the agar plate surface that does not contain medicine, continue 37 ℃ hatch 18-20 hour after, with the medicine least concentration in the corresponding test tube of not seeing bacterial growth on the agar plate as the minimum bactericidal concentration (MBC) of medicine to this bacterium.
Three, result
The Radix Isatidis eye drop all only has certain vitro antibacterial activity to the blue positive of try leather, negative bacteria.The Radix Isatidis eye drop is to staphylococcus aureus in the blue positive bacteria of try leather, staphylococcus epidermidis has certain vitro antibacterial activity, to the MIC value scope of institute's ensaying Portugal bacteria strain is that (experimental example 10 is measured the minimum adenosine 0.01875mg/ml～0.075mg) of containing in every ml eye drop to 62.5-＞250mg crude drug/ml, MIC value to taking temperature Portugal bacteria strain is＞250mg crude drug/ml, to escherichia coli in the blue negative bacterium of try leather, the ESBLs escherichia coli, Klebsiella Pneumoniae, the ESBLs Klebsiella Pneumoniae, Pseudomonas stutzeri, aerobacteria, enterobacter cloacae, the MIC value scope of husky thunder bacterium is＞250mg crude drug/ml, and the MIC value scope of examination acinetobacter calcoaceticus is 125-＞250mg crude drug/ml.
The bright Radix Isatidis eye drop of the extracorporeal disinfecting test card of Radix Isatidis eye drop does not have bactericidal action to the blue positive of try leather, negative bacteria.The Radix Isatidis eye drop to the bacteriocidal concentration of examination escherichia coli, ESBLs escherichia coli, staphylococcus aureus, Klebsiella Pneumoniae and acinetobacter calcoaceticus all greater than 250mg crude drug/ml.
The body outer disinfecting activity of table 3 Radix Isatidis eye drop
| Antibacterial (numbering) | MIC value (mg/ml) | MBC value (mg/ml) |
| Escherichia coli 0216 | 250 | ＞250 |
| ESBLs large intestine 02-3 | ＞250 | ＞250 |
| Staphylococcus aureus 02222 | 62.5 | ＞250 |
| Staphylococcus aureus 02216 | 62.5 | ＞250 |
| Lung gram 02-9-7 | ＞250 | ＞250 |
| Motionless 02511 | 125 | ＞250 |
Annotate: minimum inhibitory concentration is that 62.5mg medical material/ml (adopting the method for experimental example 10 to measure the minimum adenosine 0.01875mg/ml that contains in every ml eye drop) promptly has bacteriostasis. mention in the in-vitro antibacterial test＞the 250mg crude drug/ml explanation Radix Isatidis eye drop do not have minimum bactericidal concentration, no bactericidal action is described.But concentration is 250mg medical material/ml is fungistatic effect strong (every ml contains adenosine 0.075mg), with Mlc is that 62.5mg medical material/ml compares, P＜0.05 possesses significant difference, illustrates that concentration is that 250mg medical material/ml is that fungistatic effect strong (every ml contains adenosine 0.075mg) has stronger bacteriostasis.
Above-mentioned description of test: the Radix Isatidis eye drop only has certain vitro antibacterial activity, no bactericidal action to the blue positive of clinical isolating great majority leather, negative bacteria.
Though adenosine is not the effective ingredient of Radix Isatidis preparation, the content of adenosine can react the drug effect of Radix Isatidis preparation from the side, therefore controls the purpose that adenosine content can reach control of quality.Adenosine content can be known its MIC value by inference less than 250mg medical material/ml less than 0.075mg/ml in the Radix Isatidis eye drop, and bacteriostasis efficacy is poor.Illustrate in the control Radix Isatidis preparation that the content range of adenosine can be controlled the drug effect of Radix Isatidis preparation from the side at 0.075～2mg in every preparation unit.
The experiment of embodiment 2 Radix Isatidis eye drop extracorporeal antivirus effect experimentatioies
The Radix Isatidis eye drop (by experimental example 2 methods preparations, is measured every ml by experiment 10 methods and contained that the adenosine scope is 0.0998～2mg), the former dosage form Radix Isatidis injection (approval number of the drug: the accurate word Z14021016 of ZZ-5660 traditional Chinese medicines; Specification: 2ml/ props up, and contains crude drug 0.5g/ml, lot number: 200303267; Shanxi company of Jin Xin Double-Crane Pharmaceutical Co., Ltd produces, and purchases the western medical group company in Chengdu), substrate liquid (being colourless transparent liquid, self-control) and positive control drug aciclovir eye drop (aciclovir eye drop: the accurate word H 42020646 of traditional Chinese medicines; 8ml/ props up; Specification: 8ml:8mg; Lot number is 2003101; Produce by Hubei Qianjiang Pharmaceutical Co., Ltd..Purchase western medical group company in Chengdu) stock solution and 1: 10-1: 80 dilute liquid medicines, inoculate the Hep-2 cell culture respectively and observe.The stock solution and 1 of proof Radix Isatidis eye drop, former dosage form Radix Isatidis injection, substrate liquid and positive control drug aciclovir eye drop: 10-1: (the Hep-2 cell is that substrate detects antinuclear antibody to 80 dilute liquid medicines to the Hep-2 cell.The Hep-2 cell is provided by the eternal brightness in Shantou City bio-engineering corporation, and growth nutrient solution is 10% calf serum Eagles liquid) there is not any toxic reaction.
With adenovirus (AdV) 3 types, 4 types, 7 types, 8 types and herpes simplex virus (HSV) 1 type (providing) by Shoudu Inst. of Pediatrics.Stock solution and 1 with Radix Isatidis eye drop, former dosage form Radix Isatidis injection, substrate liquid and positive control drug aciclovir eye drop: 10-1: 80 dilute liquid medicine mixed in equal amounts effects are inoculated the Hep-2 cell culture and are observed after 1 hour.
The result shows:
The stock solution of Radix Isatidis eye drop and former dosage form Radix Isatidis injection (1: 1) all has pathological changes effect in the cell of inhibition to AdV8 type and HSV1 type virus; To pathological changes effect in AdV3,7, the 4 types virus unrestraint cell.Positive control drug, aciclovir eye drop stock solution (1: 1) all has pathological changes effect in the cell of inhibition to AdV8 and HSV1 type virus.Effect to pathological changes in AdV3,7, the equal unrestraint cell of 4 types virus.Substrate liquid and blank (Hanks liquid) are to the effect of pathological changes in AdV3,7,4,8 types and the equal unrestraint cell of HSV1 type virus.Above-mentioned experimental results show that, the Radix Isatidis eye drop (is prepared by experimental example 2 methods, experimental example 10 methods are measured every ml, and to contain the adenosine scope be in 0.0998～2mg) the scope, AdV8 type and HSV1 type virus all there is pathological changes effect in the cell of inhibition, prove absolutely, though adenosine is not an effective ingredient, can reflect the curative effect of Radix Isatidis preparation from the side, reach the purpose of control Radix Isatidis preparation by the control adenosine.
By above-mentioned experimental results show that, it is the index sexual element that Radix Isatidis preparation of the present invention adopts adenosine, to those of ordinary skill is not apparent, measure by qualitative, quantitative adenosine, can reflect Radix Isatidis medical material and stability of formulation thereof, reach the purpose of control of quality, adopting adenosine is the index sexual element, the repeatability height, stable and controllable; Prove by pharmacodynamics test, though adenosine is not an effective ingredient,, can control the curative effect of Radix Isatidis preparation from the side by control to adenosine content, make preparation more stable, controlled, for Radix Isatidis medical material and preparation thereof provide a kind of new method of quality control.
Claims (7)
1, the method for quality control of Radix Isatidis preparation is characterized in that, it is that the employing adenosine is that the index composition carries out qualitative control.
2, the method for quality control of Radix Isatidis preparation according to claim 1 is characterized in that, the method for described qualitative control is: adopting chloroform, ethyl acetate, isopropyl alcohol, water, ammonia is developing solvent, uses silica gel G F
254Lamellae launches, and reference standards is an adenosine, and under uviol lamp, wavelength is that 254nm inspects, and in the test sample chromatograph, is showing identical skin dark stain with contrast chromatograph corresponding position.
3, the method for quality control of Radix Isatidis preparation according to claim 1 is characterized in that, described preparation is eye drop, original position gel eye drop, injection, granule, capsule, oral liquid, syrup, tablet, pill or powder.
4, the method for quality control of Radix Isatidis preparation is characterized in that, it is that the employing adenosine is that the index composition is quantitatively controlled, and the content range of adenosine is 0.075～2mg in every preparation unit.
5, the method for quality control of Radix Isatidis preparation according to claim 4 is characterized in that, the method for described quantitative control is:
Chromatographic condition:
Instrument: HP1000 high performance liquid chromatograph, G1310A IsoPump pump, G1314A VMD UV-detector; The Chemstation data processing software;
Chromatographic column: Alltima C18 5 μ m, 150mm * 4.6mm;
Mobile phase: methanol-water, volume ratio are 5～12: 88～95 or phosphate buffer-methanol, and volume ratio is 15～20: 3～10; Flow velocity: 1ml/min;
Column temperature: room temperature;
Detect wavelength: 260nm;
Sample size: 20 μ l;
The preparation of need testing solution: the accurate Radix Isatidis preparation of drawing, put in the 10ml volumetric flask, be diluted to scale with 30% methanol, shake up, promptly;
The preparation of reference substance solution: precision takes by weighing the about 10mg of the adenosine that is dried to constant weight, puts in the 50ml volumetric flask, adds mobile phase and dissolves surely and be diluted to scale, shakes up, in contrast the product stock solution;
Get above solution and inject the efficient liquid phase chromatographic analysis instrument, measure, calculate by external standard method, promptly.
6, the method for quality control of Radix Isatidis preparation according to claim 5 is characterized in that, described mobile phase: methanol-water, volume ratio are 8: 92.
7, the method for quality control of Radix Isatidis preparation according to claim 4 is characterized in that, described preparation is eye drop, original position gel eye drop, injection, granule, capsule, oral liquid, syrup, tablet, pill or powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031356885A CN100352458C (en) | 2003-08-29 | 2003-08-29 | Preparaton of banlangen and method for controlling quality |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031356885A CN100352458C (en) | 2003-08-29 | 2003-08-29 | Preparaton of banlangen and method for controlling quality |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1539454A CN1539454A (en) | 2004-10-27 |
| CN100352458C true CN100352458C (en) | 2007-12-05 |
Family
ID=34323193
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB031356885A Expired - Lifetime CN100352458C (en) | 2003-08-29 | 2003-08-29 | Preparaton of banlangen and method for controlling quality |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100352458C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101403731B (en) * | 2008-10-30 | 2014-07-23 | 武汉康裕医药科技有限公司 | Isatis smalt particle content measuring method |
| CN101716215B (en) * | 2009-12-21 | 2011-08-03 | 中国科学院长春应用化学研究所 | Quality detection method of traditional Chinese medicine of isatis root |
| CN103520227A (en) * | 2013-10-09 | 2014-01-22 | 南京农业大学 | Injectable Isatis Root in-situ gel, and preparation method thereof |
| CN104297402B (en) * | 2014-07-22 | 2016-01-20 | 吉林大学 | The HPLC content assaying method of adenosine in plantain seed |
| CN105372400A (en) * | 2015-12-17 | 2016-03-02 | 江西汇仁药业有限公司 | Chinese medicine quality control method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1140597A (en) * | 1995-07-21 | 1997-01-22 | 乌鲁木齐市中亚实业有限公司 | Preparation for isatis root syrup |
| CN1141800A (en) * | 1996-04-25 | 1997-02-05 | 黎子正 | Eyedrops for treatment of cataract and production technology thereof |
| CN1277041A (en) * | 2000-04-27 | 2000-12-20 | 王洪福 | Sugar-less Radix Isatidis preparation |
| CN1318399A (en) * | 2000-04-18 | 2001-10-24 | 十堰康迪制药厂 | Radix Isatidis injecta and its preparation |
-
2003
- 2003-08-29 CN CNB031356885A patent/CN100352458C/en not_active Expired - Lifetime
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1140597A (en) * | 1995-07-21 | 1997-01-22 | 乌鲁木齐市中亚实业有限公司 | Preparation for isatis root syrup |
| CN1141800A (en) * | 1996-04-25 | 1997-02-05 | 黎子正 | Eyedrops for treatment of cataract and production technology thereof |
| CN1318399A (en) * | 2000-04-18 | 2001-10-24 | 十堰康迪制药厂 | Radix Isatidis injecta and its preparation |
| CN1277041A (en) * | 2000-04-27 | 2000-12-20 | 王洪福 | Sugar-less Radix Isatidis preparation |
Non-Patent Citations (1)
| Title |
|---|
| 《中华人民共和国药典》 519 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1539454A (en) | 2004-10-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Schipper et al. | Chitosans as absorption enhancers of poorly absorbable drugs: 3: Influence of mucus on absorption enhancement | |
| WO2015090180A1 (en) | Sanchi flower arab galactan and preparation method and use thereof | |
| CN1872106A (en) | Application of wild basil circle leaves in treating disease of virulence cold | |
| CN101181446B (en) | Application of sitsang scindapsus aureus flower as well as extract thereof in the preparation of diabetes medicament | |
| CN105287511B (en) | The application of native Radix Glycyrrhizae A | |
| CN1654481A (en) | Process for preparing glucose-reducing sand sagebrush polysaccharide and its use | |
| CN102702582A (en) | Zeamays L. polysaccharide and preparation method and application thereof | |
| CN103536635A (en) | Preparation method of holothuria nobilis and application thereof in treatment of diabetes mellitus | |
| CN100352458C (en) | Preparaton of banlangen and method for controlling quality | |
| CN109674866A (en) | A kind of anti-human primary gastrointestinal cancers pharmaceutical composition, preparation method and applications | |
| CN101524398B (en) | Medical application of licorice flavonoids | |
| CN101327247A (en) | Senna formulation and quality control method | |
| CN100574792C (en) | A kind of effective ingredient detection method of Chinese medicine composition | |
| CN1857497B (en) | Erosion treating Chinese medicine ointment and its quality control method | |
| CN101439083B (en) | Detection method of Chinese medicine soft capsules for clearing wind heat and clearing nasal passage | |
| CN103816147B (en) | The medical usage of gamlogic acid, neogambogic acid and compositions thereof | |
| CN111437263B (en) | Corncob total flavone solid dispersion osmotic pump tablet and preparation method thereof | |
| Joni et al. | The effect of lime leaf ethanol extract on diabetic nephropathy in male white rats | |
| CN1772089B (en) | Freeze dried oldenlandia powder for injection and its preparation method | |
| JP2003525839A (en) | Kinsoba buckwheat composition, production method and analysis method and use thereof | |
| CN101084951B (en) | Medicine for treating diseases concerned with respiratory and preparation method and application thereof | |
| CN101461904A (en) | Method for testing Chinese medicinal composition for treating painful swelling of throat and constipation | |
| CN100560069C (en) | A kind of Mitiglinide calcium preparation and detection method thereof | |
| CN102670698B (en) | The application of Radix Flemingiae Philippinensis extract in preparation control diabetes medicament | |
| CN101574381A (en) | Extractive of effective part of chloranthus glaber and preparation method thereof |
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 | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160105 Address after: 610000 No. 1, No. 5, building 14, nine Hing Road, Chengdu hi tech Zone, Sichuan, 903 Patentee after: Chengdu Bandung Yikang Pharmaceutical Co.,Ltd. Address before: 4, building 16, building B, Torch Hotel, No. 610041, Pioneer Road, hi tech Zone, Sichuan, Chengdu Patentee before: Chengdu Saymed Institute of Traditional Chinese Medicine |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160301 Address after: 850000 office building 110, Lhasa economic and Technological Development Zone, Tibet Autonomous Region Patentee after: Tibet Xinyi Kang Pharmaceutical Holdings Ltd. Address before: 610000 No. 1, No. 5, building 14, nine Hing Road, Chengdu hi tech Zone, Sichuan, 903 Patentee before: Chengdu Bandung Yikang Pharmaceutical Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20180814 Address after: 611630 301 industrial North Road, Pujiang Industrial Development Zone, Pujiang County, Chengdu, Sichuan Patentee after: Sichuan Heyi Pharmaceutical Co.,Ltd. Address before: 850000 110 office building, Lhasa Municipal Economic and Technological Development Zone, Tibet Autonomous Region Patentee before: Tibet Xinyi Kang Pharmaceutical Holdings Ltd. |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Isatis root preparation and quality control method thereof Effective date of registration: 20200430 Granted publication date: 20071205 Pledgee: Bank of Chengdu science and technology branch of Limited by Share Ltd. Pledgor: Sichuan Heyi Pharmaceutical Co.,Ltd. Registration number: Y2020510000045 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20210802 Granted publication date: 20071205 Pledgee: Bank of Chengdu science and technology branch of Limited by Share Ltd. Pledgor: Sichuan Heyi Pharmaceutical Co.,Ltd. Registration number: Y2020510000045 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Radix Isatidis preparation and its quality control method Effective date of registration: 20220624 Granted publication date: 20071205 Pledgee: Zhejiang Mintai commercial bank Limited by Share Ltd. Chengdu Gaoxin Branch Pledgor: Sichuan Heyi Pharmaceutical Co.,Ltd. Registration number: Y2022510000171 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| CX01 | Expiry of patent term |
Granted publication date: 20071205 |
|
| CX01 | Expiry of patent term | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20231107 Granted publication date: 20071205 Pledgee: Zhejiang Mintai commercial bank Limited by Share Ltd. Chengdu Gaoxin Branch Pledgor: Sichuan Heyi Pharmaceutical Co.,Ltd. Registration number: Y2022510000171 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right |