CN102670670A - Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones - Google Patents
Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones Download PDFInfo
- Publication number
- CN102670670A CN102670670A CN2012101780177A CN201210178017A CN102670670A CN 102670670 A CN102670670 A CN 102670670A CN 2012101780177 A CN2012101780177 A CN 2012101780177A CN 201210178017 A CN201210178017 A CN 201210178017A CN 102670670 A CN102670670 A CN 102670670A
- Authority
- CN
- China
- Prior art keywords
- ginkgo
- extract
- folium ginkgo
- dipyridamole
- injection
- 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
- 235000008100 Ginkgo biloba Nutrition 0.000 title claims abstract description 195
- 241000218628 Ginkgo Species 0.000 title claims abstract description 189
- 235000011201 Ginkgo Nutrition 0.000 title claims abstract description 189
- -1 terpene lactones Chemical class 0.000 title claims abstract description 74
- 238000002347 injection Methods 0.000 title claims abstract description 69
- 239000007924 injection Substances 0.000 title claims abstract description 69
- 235000007586 terpenes Nutrition 0.000 title claims abstract description 65
- 238000002360 preparation method Methods 0.000 title abstract description 28
- 239000000284 extract Substances 0.000 claims abstract description 99
- IZEKFCXSFNUWAM-UHFFFAOYSA-N dipyridamole Chemical compound C=12N=C(N(CCO)CCO)N=C(N3CCCCC3)C2=NC(N(CCO)CCO)=NC=1N1CCCCC1 IZEKFCXSFNUWAM-UHFFFAOYSA-N 0.000 claims abstract description 84
- 229960002768 dipyridamole Drugs 0.000 claims abstract description 84
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 70
- 238000001914 filtration Methods 0.000 claims abstract description 24
- 238000012545 processing Methods 0.000 claims abstract description 5
- 239000000463 material Substances 0.000 claims abstract description 4
- MOLPUWBMSBJXER-YDGSQGCISA-N bilobalide Chemical compound O([C@H]1OC2=O)C(=O)[C@H](O)[C@@]11[C@@](C(C)(C)C)(O)C[C@H]3[C@@]21CC(=O)O3 MOLPUWBMSBJXER-YDGSQGCISA-N 0.000 claims description 60
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 58
- 239000009429 Ginkgo biloba extract Substances 0.000 claims description 55
- 229930003944 flavone Natural products 0.000 claims description 45
- 235000011949 flavones Nutrition 0.000 claims description 45
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 42
- 210000000582 semen Anatomy 0.000 claims description 42
- 239000000243 solution Substances 0.000 claims description 42
- 238000000034 method Methods 0.000 claims description 35
- 239000000287 crude extract Substances 0.000 claims description 34
- 238000000605 extraction Methods 0.000 claims description 32
- 239000008213 purified water Substances 0.000 claims description 32
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 claims description 24
- 239000012141 concentrate Substances 0.000 claims description 24
- 150000002212 flavone derivatives Chemical class 0.000 claims description 24
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims description 24
- 238000003756 stirring Methods 0.000 claims description 22
- 150000002213 flavones Chemical class 0.000 claims description 21
- 239000007788 liquid Substances 0.000 claims description 21
- 238000010992 reflux Methods 0.000 claims description 21
- 229960004756 ethanol Drugs 0.000 claims description 19
- 239000003960 organic solvent Substances 0.000 claims description 19
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 18
- 239000003480 eluent Substances 0.000 claims description 18
- 238000001035 drying Methods 0.000 claims description 16
- 238000003810 ethyl acetate extraction Methods 0.000 claims description 14
- 238000003860 storage Methods 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- 230000001476 alcoholic effect Effects 0.000 claims description 12
- 238000001816 cooling Methods 0.000 claims description 12
- 239000006071 cream Substances 0.000 claims description 12
- 230000006837 decompression Effects 0.000 claims description 12
- 230000008021 deposition Effects 0.000 claims description 12
- 239000003814 drug Substances 0.000 claims description 12
- 239000000706 filtrate Substances 0.000 claims description 12
- 239000000796 flavoring agent Substances 0.000 claims description 12
- 235000019634 flavors Nutrition 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 12
- 150000002596 lactones Chemical class 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- 230000001954 sterilising effect Effects 0.000 claims description 12
- 238000004659 sterilization and disinfection Methods 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 239000011347 resin Substances 0.000 claims description 11
- 229920005989 resin Polymers 0.000 claims description 11
- 238000003556 assay Methods 0.000 claims description 9
- 238000010828 elution Methods 0.000 claims description 9
- 239000004952 Polyamide Substances 0.000 claims description 7
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 7
- 239000012567 medical material Substances 0.000 claims description 7
- 229920002647 polyamide Polymers 0.000 claims description 7
- 238000011084 recovery Methods 0.000 claims description 7
- 238000000638 solvent extraction Methods 0.000 claims description 7
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- 235000009827 Prunus armeniaca Nutrition 0.000 claims description 6
- 244000018633 Prunus armeniaca Species 0.000 claims description 6
- 229930003268 Vitamin C Natural products 0.000 claims description 6
- 238000010521 absorption reaction Methods 0.000 claims description 6
- 230000000274 adsorptive effect Effects 0.000 claims description 6
- 238000002425 crystallisation Methods 0.000 claims description 6
- 230000008025 crystallization Effects 0.000 claims description 6
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 6
- 238000011082 depyrogenation Methods 0.000 claims description 6
- 150000002576 ketones Chemical group 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 238000005374 membrane filtration Methods 0.000 claims description 6
- 239000012982 microporous membrane Substances 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 238000000108 ultra-filtration Methods 0.000 claims description 6
- 238000001291 vacuum drying Methods 0.000 claims description 6
- 235000019154 vitamin C Nutrition 0.000 claims description 6
- 239000011718 vitamin C Substances 0.000 claims description 6
- 239000008215 water for injection Substances 0.000 claims description 6
- 239000002671 adjuvant Substances 0.000 claims description 4
- SHWNNYZBHZIQQV-UHFFFAOYSA-J EDTA monocalcium diisodium salt Chemical compound [Na+].[Na+].[Ca+2].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O SHWNNYZBHZIQQV-UHFFFAOYSA-J 0.000 claims description 2
- 229920002565 Polyethylene Glycol 400 Polymers 0.000 claims description 2
- 239000003463 adsorbent Substances 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 238000009472 formulation Methods 0.000 claims description 2
- 235000011187 glycerol Nutrition 0.000 claims description 2
- JLFNLZLINWHATN-UHFFFAOYSA-N pentaethylene glycol Chemical compound OCCOCCOCCOCCOCCO JLFNLZLINWHATN-UHFFFAOYSA-N 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 235000013772 propylene glycol Nutrition 0.000 claims description 2
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 claims description 2
- 235000010262 sodium metabisulphite Nutrition 0.000 claims description 2
- 229930003935 flavonoid Natural products 0.000 abstract 3
- 150000002215 flavonoids Chemical class 0.000 abstract 3
- 235000017173 flavonoids Nutrition 0.000 abstract 3
- 238000005516 engineering process Methods 0.000 description 20
- 238000012360 testing method Methods 0.000 description 18
- 239000002994 raw material Substances 0.000 description 8
- 239000013558 reference substance Substances 0.000 description 8
- 244000194101 Ginkgo biloba Species 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 235000021050 feed intake Nutrition 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 239000013618 particulate matter Substances 0.000 description 4
- 229920000136 polysorbate Polymers 0.000 description 4
- 238000010257 thawing Methods 0.000 description 4
- 206010002383 Angina Pectoris Diseases 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- 206010061876 Obstruction Diseases 0.000 description 3
- 230000001154 acute effect Effects 0.000 description 3
- 230000000875 corresponding effect Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000008719 thickening Effects 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 208000006673 asthma Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 206010008118 cerebral infarction Diseases 0.000 description 2
- 208000026106 cerebrovascular disease Diseases 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 208000029078 coronary artery disease Diseases 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000000105 evaporative light scattering detection Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229940111260 ginkgo folium Drugs 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000000302 ischemic effect Effects 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 208000010125 myocardial infarction Diseases 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 201000006474 Brain Ischemia Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010008120 Cerebral ischaemia Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 208000028399 Critical Illness Diseases 0.000 description 1
- 101000783577 Dendroaspis angusticeps Thrombostatin Proteins 0.000 description 1
- 101000783578 Dendroaspis jamesoni kaimosae Dendroaspin Proteins 0.000 description 1
- 208000035126 Facies Diseases 0.000 description 1
- 241000218791 Ginkgoaceae Species 0.000 description 1
- 206010019468 Hemiplegia Diseases 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 229940127218 antiplatelet drug Drugs 0.000 description 1
- 235000019636 bitter flavor Nutrition 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000036770 blood supply Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 208000009973 brain hypoxia - ischemia Diseases 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000000916 dilatatory effect Effects 0.000 description 1
- 230000010339 dilation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 201000005577 familial hyperlipidemia Diseases 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000006386 memory function Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 208000031225 myocardial ischemia Diseases 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 238000009512 pharmaceutical packaging Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 239000000106 platelet aggregation inhibitor Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000003405 preventing effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 239000003848 thrombocyte activating factor antagonist Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones. The preparation method comprises the following steps: 1), collecting, processing, detecting and classifying ginkgo leaves; 2), extracting and preparing ginkgo terpene lactones extract from ginkgo leaves; 3), extracting ginkgo leaf flavonoid extract; 4), reflowing ginkgo leaf flavonoid extract and partial dipyridamole by adding water, and then filtering so as to be prepared into mixed liquor; 5), mixing the mixed liquor, ginkgo terpene lactones extract, the rest dipyridamole and auxiliary material to be prepared into ginkgo dipyridolum injection as per conventional preparation method; and in the ginkgo dipyridolum injection, the content of the total ginkgo leaf flavonoid and that of the dipyridamole conform to the conventional requirements, and the content of the ginkgo terpene lactones ranges from 0.5 to 1.2 mg/mL.
Description
Technical field
The present invention relates to a kind of method for preparing of injection, specifically, relate to the method for preparing of the higher Ginkgo Leaf Extract and Dipyridamole Injection of a kind of ginkalide A, ginkalide B, ginkalide C and content of bilobalide.
Background technology
Folium Ginkgo and dipyridamole are the main initiation materials of producing Ginkgo Leaf Extract and Dipyridamole Injection.Wherein, The dried leaves of Semen Ginkgo leaf system Ginkgoaceae plant Ginkgo biloba Ginkgo biloba L.; Have blood circulation promoting and blood stasis dispelling, removing obstruction in the collateral to relieve pain, astringe the lung and relieving asthma, change turbid function of lowering fat, clinically be used for diseases such as obstruction of collaterals by blood stasis, obstruction of qi in the chest and cardialgia, apoplectic hemiplegia, cough and asthma due to lung deficiency and hyperlipidemia.The complex chemical composition of Folium Ginkgo mainly comprises flavone, lactone, terpenoid, phenolic acid, macromolecule, alkaloid, polysaccharide, aminoacid, trace element etc.The chemistry of dipyridamole (Dipyridamole) by name 2,2 ', 2 ", 2 " '-[4,8-dipiperidino pyrimido [5,4-d] pyrimidine-2,6-two bases] two nitrilo-s]-tetraethoxide, be yellow crystalline powder; 163 ℃ of fusing points; Be soluble in chloroform and diluted acid, be dissolved in ethanol, be slightly soluble in acetone, water-soluble hardly; Odorless, mildly bitter flavor.Clinically, dipyridamole is used to treat angina pectoris and myocardial infarction mainly as coronary dilation agent and platelet aggregation inhibitor, recurrence of prevention myocardial infarction and angina pectoris.Ginkgo Leaf Extract and Dipyridamole Injection is a compound injection product that is prepared from Folium Ginkgo extract and chemical synthetic drug dipyridamole.
This pharmaceutical composition has the removing interior free yl, the not effect of receptor 1 activity radical damage of protection body.Clinically, Ginkgo Leaf Extract and Dipyridamole Injection is applicable to prevention and treats coronary heart disease, thrombotic disease, is bringing into play important effect in the anxious critical illness of treatment field.Contained Ginkgo total flavones has dilating coronary blood vessel, cerebrovascular in the Ginkgo Leaf Extract and Dipyridamole Injection; Improve symptoms of cerebral ischemia and memory function; Dipyridamole ability anticoagulant; High concentration can suppress platelet and discharge, and for acute and chronic ischemic cerebrovascular prevention and therapeutical effect is preferably arranged all.Report according to clinical literature; Ginkgo Leaf Extract and Dipyridamole Injection all has good therapeutical effect to the acute and chronic ischemic cerebrovascular, and can significantly improve hemorheology index, reduces blood viscosity; Promoting the recovery of function of nervous system, is one of ideal medicament of treatment acute cerebral infarction.In addition, the effect that Ginkgo Leaf Extract and Dipyridamole Injection is transferred fat in addition, improved hemorheology and improve blood supply of cardiac muscle, be treatment aged coronary heart disease patient a kind of effectively and safe drugs also can be used for treating cardiovascular and cerebrovascular disease and merge hyperlipemia and high sticky blood.
Bilobalide has anticoagulation, prevents and treats atherosclerosis, protects many pharmacological actions such as ischemia damaged tissue, shock, antiinflammatory as the extract of Folium Ginkgo; Can effectively suppress the inductive platelet aggregation of multiple derivant; And bilobalide has been considered to efficient special PAF antagonist, has inhibition platelet activating factor-PAF and to the protective effect of myocardial ischemia, cerebral hypoxia ischemia.
But in the existing production technology of Ginkgo Leaf Extract and Dipyridamole Injection, most of technologies are only followed the tracks of the flavones content of sample, and fail the terpene lactone composition is monitored; In the existing production technology, all be that Folium Ginkgo is extracted with ethanol or water, absorption then; Eluting for example obtains ginkgo biloba succi:
CN 1939357 discloses the Folium Ginkgo extraction process: after the present invention pulverizes Folium Ginkgo, and through alcohol reflux, thickening filtration, resin absorption, technical finesses such as ethanol elution obtain ginkgo biloba succi.
CN 101947247B discloses the Folium Ginkgo extraction process, then adopts purified water extraction.
We have tested said method, and the terpene lactone component content is very low in the ginkgo biloba succi.
Terpene lactone constituents in the Folium Ginkgo mainly contains ginkalide A, ginkalide B, ginkalide C, bilobalide J and bilobalide; This constituents receives the restriction of conditions such as the medical material place of production, collecting time, processing and storage; Secondly, Different Extraction Method also has considerable influence to the content of terpene lactone.Therefore, a plurality of factors and index have influence on the terpene lactone contents in the Ginkgo Leaf Extract and Dipyridamole Injection, if do not add attention, with the curative effect that has a strong impact on product, finally bring security risks for clinical application.
Summary of the invention
The object of the present invention is to provide a kind of method for preparing of Ginkgo Leaf Extract and Dipyridamole Injection of high Folium Ginkgo terpene lactones content; This method can improve the content of ginkalide A, ginkalide B, ginkalide C and bilobalide in Folium Ginkgo extract and the Ginkgo Leaf Extract and Dipyridamole Injection significantly, improves the clinical efficacy and the quality stability of Ginkgo Leaf Extract and Dipyridamole Injection.
In order to realize the object of the invention, the method for preparing of Ginkgo Leaf Extract and Dipyridamole Injection of the present invention, it comprises the steps:
1) collection of Folium Ginkgo, processing, detection and classification;
2) from Folium Ginkgo, extract Folium Ginkgo terpene lactone extract;
3) extract the Folium Ginkgo flavone extract;
4) will contain Semen Ginkgo total flavones 10g the Folium Ginkgo flavone extract and 2 the gram dipyridamoles add water backflow after-filtration, process mixed liquor 1;
5) above-mentioned full dose mixed liquor 1, the Folium Ginkgo terpene lactone extract that contains Ginkgo total lactones 5g, 2 gram dipyridamoles and adjuvant are mixed; Process Ginkgo Leaf Extract and Dipyridamole Injection by the conventional formulation method; In the said Ginkgo Leaf Extract and Dipyridamole Injection; The content of Ginkgo total flavones and dipyridamole is except that should meeting conventional the requirement, and the content of Semen Ginkgo terpene lactone should be 0.5～1.2mg/mL.
Above-mentioned method for preparing; Step 1) is specially: be that late June is to late October the collection period of Folium Ginkgo; Bright leaf at source in carrying out drying below 60 ℃, is selected for use the total content of ginkalide A, ginkalide B, ginkalide C and bilobalide to be higher than 0.35% Folium Ginkgo and extracted.
Above-mentioned method for preparing, step 2) is specially: Folium Ginkgo is put in the extraction element, added 40～80% organic solvent of 7～15 times of amount medical material weight, heating and refluxing extraction; Wherein, organic solvent can be selected any one in methanol, ethanol, acetone, the acetonitrile for use, extracts 1～3 hour; Extract merge extractive liquid,, decompression and solvent recovery under the low temperature altogether 1～3 time; Concentrate drying makes the organic solvent crude extract of Folium Ginkgo, stays the Folium Ginkgo medicinal residues after the extraction.Crude extract adds ethyl acetate, extracts after-filtration, filtrated and with ethyl acetate extraction after crude extract 2.Merging filtrate, decompression filtrating is reclaimed ethyl acetate, adds 80～100% alcoholic solution, makes abundant dissolving, filters; In the polyamide chromatographic column of having handled well, carry out eluting on the filtrating, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with 80～100% alcoholic solution; In alumina column, carry out eluting on the concentrated solution, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with dehydrated alcohol; Crystallization is filtered, and drying promptly gets Folium Ginkgo terpene lactone extract.
Above-mentioned method for preparing, step 3) is specially: with Folium Ginkgo medicinal residues behind the organic solvent extraction and the crude extract 2 behind the ethyl acetate extraction, add 7～15 times purified water of medicinal residues and crude extract 2 gross weights; Heating and refluxing extraction was extracted 1～3 hour, extracted altogether 1～3 time; Merge extractive liquid, is evaporated to 1/16～1/8 of original volume, with concentrated solution placement, deposition, centrifugal, filtration; Filtrating gone up adsorb, wherein, can select any among macroporous adsorbent resin AB-8, ADS-17, D101, DM130, D201, HPD417, HPD600, HPD722, HZ803, HZ806, the HZ841 for use in the macroporous adsorptive resins of having handled well; Extracting solution with the purified water washing of 8～12 times of column volume amounts, is used ethanol elution then after fully adsorbing; Collect the apricot yellow ketone part of argentiferous eluent, be concentrated into and do not have the alcohol flavor, add the purified water of 0.5～2 times of Folium Ginkgo weight then, stir leave standstill after the cooling, deposition, centrifugal, filter; To filtrate to concentrate and receive cream; Be dried to water content below 6%, pulverize, promptly get the Semen Ginkgo water extract.
The concentration expressed in percentage by volume that adds 8～12 times of weight in the Folium Ginkgo water extract is the ethanol more than 90%, is stirred to dissolving, filters; Filtrating is concentrated into nothing alcohol flavor, adds 10 times of purified water that concentrate the back medicine liquid volumes then, leaves standstill after the stirring cooling; Get supernatant, after centrifugalize, concentrate and receive cream, vacuum drying to water content is below 6%; Pulverize, promptly get the Folium Ginkgo flavone extract.
Above-mentioned method for preparing, step 4) is specially: get the purified water that the Folium Ginkgo flavone extract and 2 that contains Semen Ginkgo total flavones 10g restrains dipyridamoles and 10～15 times of weight of extract weight, fully stir mix homogeneously; Adjust pH to 3.5～5.5, reflux 0.5～1 hour is cooled to room temperature; Supernatant is got in readjustment pH value to 3.5～5.5, with the membrane filtration of 0.2mm; Filtrating is the ultrafilter membrane ultrafiltration below 100,000 dalton again, the filtrating autoclave sterilization; Put coldly, promptly get and process mixed liquor 1, low tempertaure storage.
Above-mentioned method for preparing is characterized in that, step 5) is specially: with above-mentioned full dose mixed liquor 1, the Folium Ginkgo terpene lactone extract that contains Ginkgo total lactones 5g, 2 gram dipyridamole and relevant auxiliary materials, dissolve fully with water for injection; Wherein, adjuvant is one or more in propylene glycol, glycerol, Tween 80, sodium pyrosulfite, vitamin C, calcium disodium edetate or the PEG400, and adjust pH is 3.5～5.5, and deepfreeze is placed; Filter, depyrogenation makes the injection intermediate; Through after the assay was approved, use the 0.22mm filtering with microporous membrane, the filtrating fill; Autoclave sterilization promptly gets Ginkgo Leaf Extract and Dipyridamole Injection, low tempertaure storage.
The invention has the advantages that: the method for preparing of Ginkgo Leaf Extract and Dipyridamole Injection of the present invention; Feed intake through preferred high lactone content Folium Ginkgo raw material; Carrying the certain density organic solvent of process using extracts; Adopt organic solvent, polyamide column and alumina column to carry out enrichment then; Improved the extraction ratio of Semen Ginkgo Folium Ginkgo terpene lactone, technology of the present invention and CN 1939357, the disclosed Folium Ginkgo extraction process of CN 101947247B compare, and the extraction ratio of its Semen Ginkgo Folium Ginkgo terpene lactone is much higher than the latter two technologies.Its beneficial effect is seen embodiment 5.
The present invention be advantageous in that: the Folium Ginkgo terpene lactone was added in the injection in the preparation stage, can avoid strong acid and strong base destruction to the lactone composition in the long-time course of processing of intermediate effectively.
Therefore; The finished product that makes is except that the stable content of total flavones and dipyridamole is controlled; The content of Semen Ginkgo terpene lactones such as ginkalide A, ginkalide B, ginkalide C, bilobalide has also reached stable and controllable; Improve Ginkgo Leaf Extract and Dipyridamole Injection quality of stability and controllability, guaranteed the effectiveness of clinical application.Its beneficial effect is seen embodiment 8.
Description of drawings
The chromatogram of Fig. 1 Semen Ginkgo terpene lactone reference substance, among the figure, the 1-bilobalide; The 2-ginkalide C; The 3-ginkalide A; The 4-bilobalide;
The chromatogram of Fig. 2 Ginkgo Leaf Extract and Dipyridamole Injection terpene lactone, among the figure, the 1-bilobalide; The 2-ginkalide C; The 3-ginkalide A; The 4-bilobalide;
The chromatogram of Fig. 3 Semen Ginkgo extrac negative sample.
The specific embodiment
Below further specify the present invention through specific embodiment, but be not used for limiting scope of the present invention.
Embodiment 1
1, Folium Ginkgo raw material: Folium Ginkgo picks up from the leaf of the Ginkgoaceae plant Ginkgo biloba of artificial growth; The age of tree was generally 6～15 years, and be mid-June collection period, and bright leaf is at source carried out forced air drying in about 60 ℃; Moisture content is controlled at below 6%; Store in aeration-drying place the good back of packing, and the total content of measuring ginkalide A, ginkalide B, ginkalide C and bilobalide in the Folium Ginkgo is 0.361%, and measuring content of total flavone is 0.897%.
2, Folium Ginkgo terpene lactone extract: Folium Ginkgo is put in the extraction element,, added concentration and be 70% alcoholic solution by feed intake 10 times of weight of medical material; Heating and refluxing extraction was extracted 2 hours, extracted secondary altogether; Merge extractive liquid,, decompression and solvent recovery under the low temperature, concentrate drying; Make the organic solvent crude extract of Folium Ginkgo, stay the Folium Ginkgo medicinal residues after the extraction.Crude extract adds the ethyl acetate of 3 times of amounts of crude extract, extracts after-filtration, filtrated and with ethyl acetate extraction after crude extract 2.Merging filtrate, decompression filtrating is reclaimed ethyl acetate, adds 80% alcoholic solution, makes abundant dissolving, filters; In the polyamide chromatographic column of having handled well, carry out eluting on the filtrating, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with 80% alcoholic solution; In alumina column, carry out eluting on the concentrated solution, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with dehydrated alcohol; Crystallization is filtered, and drying promptly gets Folium Ginkgo terpene lactone extract.
3, Folium Ginkgo flavone extract: with Folium Ginkgo medicinal residues behind the organic solvent extraction and the crude extract 2 behind the ethyl acetate extraction, add 7 times purified water of medicinal residues and crude extract 2 gross weights, heating and refluxing extraction was extracted 3 hours; Extract altogether 3 times, merge extractive liquid, is evaporated to 1/16 of original volume; With concentrated solution placement, deposition, centrifugal, filtration, filtrating going up adsorbed in the HZ841 macroporous adsorptive resins of having handled well, extracting solution is after fully adsorbing; With the washing of the purified water of 8 times of column volume amounts, use ethanol elution then, collect the apricot yellow ketone part of argentiferous eluent, be concentrated into and do not have the alcohol flavor; The purified water that adds 0.5 times of Folium Ginkgo weight then, stir leave standstill after the cooling, deposition, centrifugal, filter, will filtrate concentrates receipts cream; Be dried to water content below 6%, pulverize, promptly get the Semen Ginkgo water extract.
The concentration expressed in percentage by volume that adds 8 times of weight in the Folium Ginkgo water extract is the ethanol more than 90%, is stirred to dissolving, filters; Filtrating is concentrated into nothing alcohol flavor, adds 10 times of purified water that concentrate the back medicine liquid volumes then, leaves standstill after the stirring cooling; Get supernatant, after centrifugalize, concentrate and receive cream, vacuum drying to water content is below 6%; Pulverize, promptly get the Folium Ginkgo flavone extract.
4, the preparation of Ginkgo Leaf Extract and Dipyridamole Injection:
1) prescription is formed
Annotate: by Ginkgo total flavones: 10g, show the content of the before measurement Folium Ginkgo flavone extract that feeds intake, the amount that contains the Folium Ginkgo flavone extract of 10g is an inventory.
By Ginkgo total lactones: 5g, show the content of the before measurement Semen Ginkgo terpenoid lactone extract that feeds intake, the amount that contains the Semen Ginkgo terpenoid lactone extract of 5g is an inventory.
2) preparation technology
A: process mixed liquor 1: get the Folium Ginkgo flavone extract (by Ginkgo total flavones: 10g), add the dipyridamole and the purified water of 10 times of weight of extract weight of extract weight 2g, fully stirring, mix homogeneously; Adjust pH to 3.5～5.5, reflux 0.5 hour is cooled to room temperature; Supernatant is got in readjustment pH value to 3.5～5.5, with the membrane filtration of 0.2mm; Filtrating is the ultrafilter membrane ultrafiltration below 100,000 dalton again, the filtrating autoclave sterilization; Put coldly, promptly get mixed liquor 1, low tempertaure storage;
B: (by Ginkgo total lactones: 5g), 2g dipyridamole and 20g Tween 80,50g vitamin C, 1000ml propylene glycol, the water for injection 8000ml of residue dosage stir dissolving fully, adjust pH is 3.5～5.5, and deepfreeze is placed with above-mentioned full dose mixed liquor 1, Folium Ginkgo terpene lactone extract; Filter, filtrating is supplied 10000ml, depyrogenation, makes the injection intermediate; Through after the assay was approved; Use the 0.22mm filtering with microporous membrane, filtrating fill, autoclave sterilization; Promptly get Ginkgo Leaf Extract and Dipyridamole Injection, low tempertaure storage.
1, Folium Ginkgo raw material: with embodiment 1.
2, Folium Ginkgo terpene lactone extract: Folium Ginkgo is put in the extraction element,, added concentration and be 80% methanol solution by feed intake 7 times of weight of medical material; Heating and refluxing extraction was extracted 3 hours, extracted altogether 3 times; Merge extractive liquid,, decompression and solvent recovery under the low temperature, concentrate drying; Make the organic solvent crude extract of Folium Ginkgo, stay the Folium Ginkgo medicinal residues after the extraction.Crude extract adds the ethyl acetate of 4 times of amounts of crude extract, extracts after-filtration, filtrated and with ethyl acetate extraction after crude extract 2.Merging filtrate, decompression filtrating is reclaimed ethyl acetate, adds 90% alcoholic solution, makes abundant dissolving, filters; In the polyamide chromatographic column of having handled well, carry out eluting on the filtrating, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with 90% alcoholic solution; In alumina column, carry out eluting on the concentrated solution, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with dehydrated alcohol; Crystallization is filtered, and drying promptly gets Folium Ginkgo terpene lactone extract.
3, Folium Ginkgo flavone extract: with Folium Ginkgo medicinal residues behind the organic solvent extraction and the crude extract 2 behind the ethyl acetate extraction, add 15 times purified water of medicinal residues and crude extract 2 gross weights, heating and refluxing extraction was extracted 3 hours; Extract altogether 3 times, merge extractive liquid, is evaporated to 1/8 of original volume; With concentrated solution placement, deposition, centrifugal, filtration, filtrating going up adsorbed in the AB-8 macroporous adsorptive resins of having handled well, extracting solution is after fully adsorbing; With the washing of the purified water of 12 volumes, use ethanol elution then, collect the apricot yellow ketone part of argentiferous eluent, be concentrated into and do not have the alcohol flavor; The purified water that adds 2 times of Folium Ginkgo weight then, stir leave standstill after the cooling, deposition, centrifugal, filter, will filtrate concentrates receipts cream; Be dried to water content below 6%, pulverize, promptly get the Semen Ginkgo water extract.
The concentration expressed in percentage by volume that adds 12 amounts in the Folium Ginkgo water extract is the ethanol more than 90%, is stirred to dissolving, filters; Filtrating is concentrated into nothing alcohol flavor, adds 10 times of purified water that concentrate the back medicine liquid volumes then, leaves standstill after the stirring cooling; Get supernatant, after centrifugalize, concentrate and receive cream, vacuum drying to water content is below 6%; Pulverize, promptly get the Folium Ginkgo flavone extract.
4, the preparation of Ginkgo Leaf Extract and Dipyridamole Injection:
1) prescription is formed
2) preparation technology
A: process mixed liquor 1: get the Folium Ginkgo flavone extract (by Ginkgo total flavones: 10g), add the dipyridamole and the purified water of 10 times of weight of extract weight of extract weight 2g, fully stirring, mix homogeneously; Adjust pH to 3.5～5.5, reflux 0.5 hour is cooled to room temperature; Supernatant is got in readjustment pH value to 3.5～5.5, with the membrane filtration of 0.2mm; Filtrating is the ultrafilter membrane ultrafiltration below 100,000 dalton again, the filtrating autoclave sterilization; Put coldly, promptly get mixed liquor 1, low tempertaure storage;
B: (by Ginkgo total lactones: 5g), 2g dipyridamole and 20g Tween 80,50g vitamin C, 1000ml propylene glycol, the water for injection 8000ml of residue dosage stir dissolving fully, adjust pH is 3.5～5.5, and deepfreeze is placed with above-mentioned full dose mixed liquor 1, Folium Ginkgo terpene lactone extract; Filter, filtrating is supplied 10000ml, depyrogenation, makes the injection intermediate; Through after the assay was approved; Use the 0.22mm filtering with microporous membrane, filtrating fill, autoclave sterilization; Promptly get Ginkgo Leaf Extract and Dipyridamole Injection, low tempertaure storage.
Embodiment 3
1, Folium Ginkgo raw material: with embodiment 1.
2, Folium Ginkgo terpene lactone extract: Folium Ginkgo is put in the extraction element,, added concentration and be 40% acetone soln by feed intake 15 times of weight of medical material; Heating and refluxing extraction was extracted 2 hours, extracted secondary altogether; Merge extractive liquid,, decompression and solvent recovery under the low temperature, concentrate drying; Make the organic solvent crude extract of Folium Ginkgo, stay the Folium Ginkgo medicinal residues after the extraction.Crude extract adds the ethyl acetate of 3 times of amounts of crude extract, extracts after-filtration, filtrated and with ethyl acetate extraction after crude extract 2.Merging filtrate, decompression filtrating is reclaimed ethyl acetate, adds 100% alcoholic solution, makes abundant dissolving, filters; On the filtrating in the polyamide chromatographic column of having handled well, with identical (what specifically is?) solution carries out eluting, collects the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure; In alumina column, carry out eluting on the concentrated solution, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with dehydrated alcohol; Crystallization is filtered, and drying promptly gets Folium Ginkgo terpene lactone extract.
3, Folium Ginkgo flavone extract: with Folium Ginkgo medicinal residues behind the organic solvent extraction and the crude extract 2 behind the ethyl acetate extraction, add 10 times purified water of medicinal residues and crude extract 2 gross weights, heating and refluxing extraction was extracted 3 hours; Extract altogether 3 times, merge extractive liquid, is evaporated to 1/12 of original volume; With concentrated solution placement, deposition, centrifugal, filtration, filtrating going up adsorbed in the HPD722 macroporous adsorptive resins of having handled well, extracting solution is after fully adsorbing; With the washing of the purified water of 10 times of column volume amounts, use ethanol elution then, collect the apricot yellow ketone part of argentiferous eluent, be concentrated into and do not have the alcohol flavor; The purified water that adds 1 times of Folium Ginkgo weight then, stir leave standstill after the cooling, deposition, centrifugal, filter, will filtrate concentrates receipts cream; Be dried to water content below 6%, pulverize, promptly get the Semen Ginkgo water extract.
The concentration expressed in percentage by volume that adds 10 times of weight in the Folium Ginkgo water extract is the ethanol more than 90%, is stirred to dissolving, filters; Filtrating is concentrated into nothing alcohol flavor, adds 10 times of purified water that concentrate the back medicine liquid volumes then, leaves standstill after the stirring cooling; Get supernatant, after centrifugalize, concentrate and receive cream, vacuum drying to water content is below 6%; Pulverize, promptly get the Folium Ginkgo flavone extract.
4, the preparation of Ginkgo Leaf Extract and Dipyridamole Injection:
1) prescription is formed
2) preparation technology
A: process mixed liquor 1: get the Folium Ginkgo flavone extract (by Ginkgo total flavones: 10g), add the dipyridamole and the purified water of 10 times of weight of extract weight of extract weight 2g, fully stirring, mix homogeneously; Adjust pH to 3.5～5.5, reflux 0.5 hour is cooled to room temperature; Supernatant is got in readjustment pH value to 3.5～5.5, with the membrane filtration of 0.2mm; Filtrating is the ultrafilter membrane ultrafiltration below 100,000 dalton again, the filtrating autoclave sterilization; Put coldly, promptly get mixed liquor 1, low tempertaure storage;
B: (by Ginkgo total lactones: 5g), 2g dipyridamole and 20g Tween 80,50g vitamin C, 1000ml propylene glycol, the water for injection 8000ml of residue dosage stir dissolving fully, adjust pH is 3.5～5.5, and deepfreeze is placed with above-mentioned full dose mixed liquor 1, Folium Ginkgo terpene lactone extract; Filter, filtrating is supplied 10000ml, depyrogenation, makes the injection intermediate; Through after the assay was approved; Use the 0.22mm filtering with microporous membrane, filtrating fill, autoclave sterilization; Promptly get Ginkgo Leaf Extract and Dipyridamole Injection, low tempertaure storage.
Embodiment 4
1, Folium Ginkgo raw material: with embodiment 1.
2, Folium Ginkgo terpene lactone extract: Folium Ginkgo is put in the extraction element,, added concentration and be 70% acetonitrile solution by feed intake 10 times of weight of medical material; Heating and refluxing extraction was extracted 3 hours, extracted secondary altogether; Merge extractive liquid,, decompression and solvent recovery under the low temperature, concentrate drying; Make the organic solvent crude extract of Folium Ginkgo, stay the Folium Ginkgo medicinal residues after the extraction.Crude extract adds the ethyl acetate of 3 times of amounts of crude extract, extracts after-filtration, filtrated and with ethyl acetate extraction after crude extract 2.Merging filtrate, decompression filtrating is reclaimed ethyl acetate, adds 90% alcoholic solution, makes abundant dissolving, filters; In the polyamide chromatographic column of having handled well, carry out eluting on the filtrating, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with 90% alcoholic solution; In alumina column, carry out eluting on the concentrated solution, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with dehydrated alcohol; Crystallization is filtered, and drying promptly gets Folium Ginkgo terpene lactone extract.
3, Folium Ginkgo flavone extract: with Folium Ginkgo medicinal residues behind the organic solvent extraction and the crude extract 2 behind the ethyl acetate extraction, add 7 times purified water of medicinal residues and crude extract 2 gross weights, heating and refluxing extraction was extracted 3 hours; Extract altogether 3 times, merge extractive liquid, is evaporated to 1/16 of original volume; With concentrated solution placement, deposition, centrifugal, filtration, filtrating going up adsorbed in the HZ803 macroporous adsorptive resins of having handled well, extracting solution is after fully adsorbing; With the washing of the purified water of 8 times of column volume amounts, use ethanol elution then, collect the apricot yellow ketone part of argentiferous eluent, be concentrated into and do not have the alcohol flavor; The purified water that adds 0.5 times of Folium Ginkgo weight then, stir leave standstill after the cooling, deposition, centrifugal, filter, will filtrate concentrates receipts cream; Be dried to water content below 6%, pulverize, promptly get the Semen Ginkgo water extract.
The concentration expressed in percentage by volume that adds 8 times of weight in the Folium Ginkgo water extract is the ethanol more than 90%, is stirred to dissolving, filters; Filtrating is concentrated into nothing alcohol flavor, adds 10 times of purified water that concentrate the back medicine liquid volumes then, leaves standstill after the stirring cooling; Get supernatant, after centrifugalize, concentrate and receive cream, vacuum drying to water content is below 6%; Pulverize, promptly get the Folium Ginkgo flavone extract.
4, the preparation of Ginkgo Leaf Extract and Dipyridamole Injection:
1) prescription is formed
A: process mixed liquor 1: get the Folium Ginkgo flavone extract (by Ginkgo total flavones: 10g), add the dipyridamole and the purified water of 10 times of weight of extract weight of extract weight 2g, fully stirring, mix homogeneously; Adjust pH to 3.5～5.5, reflux 0.5 hour is cooled to room temperature; Supernatant is got in readjustment pH value to 3.5～5.5, with the membrane filtration of 0.2mm; Filtrating is the ultrafilter membrane ultrafiltration below 100,000 dalton again, the filtrating autoclave sterilization; Put coldly, promptly get mixed liquor 1, low tempertaure storage;
B: (by Ginkgo total lactones: 5g), 2g dipyridamole and 20g Tween 80,50g vitamin C, 1000ml propylene glycol, the water for injection 8000ml of residue dosage stir dissolving fully, adjust pH is 3.5～5.5, and deepfreeze is placed with above-mentioned full dose mixed liquor 1, Folium Ginkgo terpene lactone extract; Filter, filtrating is supplied 10000ml, depyrogenation, makes the injection intermediate; Through after the assay was approved; Use the 0.22mm filtering with microporous membrane, filtrating fill, autoclave sterilization; Promptly get Ginkgo Leaf Extract and Dipyridamole Injection, low tempertaure storage.
The comparison that embodiment 5 extracts Semen Ginkgo terpene lactone technology:
Technology A
1, Folium Ginkgo raw material: with embodiment 1,1 kilogram of inventory.
2, extract Semen Ginkgo terpene lactone technology with among the embodiment 12, extract-sample A.
Technology B
1, Folium Ginkgo raw material: with embodiment 1,1 kilogram of inventory.
2, extract Semen Ginkgo terpene lactone technology with CN 1939357, with after the Folium Ginkgo pulverizing, through alcohol reflux, thickening filtration, resin absorption, technical finesses such as ethanol elution obtain ginkgo biloba succi-sample B
Technology C
1, Folium Ginkgo raw material: with embodiment 1,1 kilogram of inventory.
2, extract the same CN101947247B of Semen Ginkgo terpene lactone technology, Folium Ginkgo pulverized after, purified water reflux, extract,, thickening filtration, resin absorption, technical finesses such as ethanol elution obtain ginkgo biloba succi-sample C.
The assay method of Semen Ginkgo terpene lactone is seen embodiment 6.
That extracts Semen Ginkgo terpene lactone technology relatively sees the following form 1:
Table 1: the mensuration result of Folium Ginkgo terpene lactones
Conclusion: can find out that from table 1 technology of the present invention and CN 1939357, the disclosed Folium Ginkgo extraction process of CN 101947247B compare, the extraction ratio of its Semen Ginkgo terpene lactone is much higher than the latter two technologies.
The mensuration of embodiment 6 Semen Ginkgo terpene lactones
One, instrument, medicine, reagent
Instrument: Agilent 1200 series of high efficiency chromatograph of liquid, Altech 2000ES evaporative light scattering detector; Reference substance: bilobalide (lot number 100865-200605), ginkalide A (lot number 100862-200608), ginkalide B (lot number 100863-200508), ginkalide C (lot number 100864-200906), all purchase in Nat'l Pharmaceutical & Biological Products Control Institute; Reagent: acetonitrile (chromatographically pure), isopropyl alcohol (chromatographically pure), water (deionized water).
Two, assay method
2.1 the preparation of reference substance solution
Get ginkalide A, ginkalide B, ginkalide C and bilobalide reference substance, precision weighing adds methanol and processes every 1ml bilobalide-containing A 0.5mg, and the mixed solution of ginkalide B, C and each 0.2mg of bilobalide promptly gets.
2.2 the preparation of need testing solution
Ginkgo Leaf Extract and Dipyridamole Injection, specification is 10ml, gets lot number and be 10 samples of 20100201, breaks glass into pieces, in the triangular flask of impouring 250ml; Mix homogeneously, precision is drawn 30ml, places the separatory funnel of 100ml, and the hydrochloric acid solution with 2% is regulated about pH to 2.0, with 4 (30ml of ethyl acetate extraction; 20ml, 20ml, 20ml), the combined ethyl acetate layer washs with 5% sodium acetate solution 20ml; Obtain sodium acetate solution, use the 10ml ethyl acetate extraction, combined ethyl acetate, with the 20ml water washing, organic facies is concentrated into dried; Residue adds acetone and is transferred in the 2ml volumetric flask, and is settled to scale with acetone, shakes up, filters, and promptly gets.
2.3 chromatographic condition
Chromatographic column: Welch C
18Chromatographic column (250 * 4.6mm, 5mm); Mobile phase: methanol-isopropyl alcohol-water (9: 7.5: 83.5); Column temperature: 30 ℃; Flow velocity: 1.0ml/min; The drift tube temperature of evaporative light scattering detector: 105 ℃, flow rate of carrier gas: 3.0L/min; Number of theoretical plate must not be lower than 3000 by bilobalide.Fig. 1, Fig. 2 and Fig. 3 are respectively the chromatogram of Semen Ginkgo terpene lactone reference substance, Ginkgo Leaf Extract and Dipyridamole Injection and Semen Ginkgo extrac negative sample.
Visible by Fig. 1, Fig. 2 and Fig. 3: the chromatogram of Ginkgo Leaf Extract and Dipyridamole Injection sample has corresponding chromatographic peak to occur in each retention time corresponding position of terpene lactone standard substance; And the Semen Ginkgo extrac negative sample does not disturb in corresponding position, explains that this chromatographic condition can be used for analyzing the content of the terpene lactone of Ginkgo Leaf Extract and Dipyridamole Injection.
2.4 linear relationship
Accurate absorption reference substance 2,4,6,8,10,15mL inject chromatograph of liquid, measure and write down chromatogram by above-mentioned chromatographic condition.Logarithm value with the quality of bilobalide, ginkalide A, ginkalide B and ginkalide C reference substance is an abscissa respectively, is vertical coordinate with the logarithm value of peak area, the drawing standard curve.Mensuration result is following:
Ginkalide A:
Standard curve: the y=1.4511x+6.6004r=0.9992 range of linearity: 3.12ug～11.73ug
Ginkalide B:
Standard curve: the y=1.4253x+6.5094r=0.9991 range of linearity: 3.13ug～11.76ug
Ginkalide C:
Standard curve: the y=1.4146x+6.6309r=0.9985 range of linearity: 3.09ug～11.62ug
Bilobalide:
Standard curve: y=1.958x+2.7728r=0.9993 linear range: 0.96ug～14.40ug
Three, sample determination
Get the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation, parallel three parts, with reference to the method for preparing of need testing solution, handle respectively, make the Ginkgo Leaf Extract and Dipyridamole Injection sample that respectively supplies the Semen Ginkgo terpene lactone contents to measure.Accurate respectively reference substance solution 10ml, the 20ml of absorbing; Need testing solution 10～20ml; Inject chromatograph of liquid respectively, measure, calculate the content of ginkalide A, ginkalide B, ginkalide C and bilobalide with external standard two-point method logarithmic equation respectively; Promptly get, each measured value is added and promptly gets the total content of Semen Ginkgo terpene lactone.The sample determination result of Ginkgo Leaf Extract and Dipyridamole Injection embodiment 1～4 sees the following form.
The assay result of table 2 Ginkgo Leaf Extract and Dipyridamole Injection terpene lactone
Visible from last table: the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation; The content of Folium Ginkgo terpene lactones is 0.5～1.2mg/ml; Basically can control the content of the Folium Ginkgo terpene lactones in the preparation finished product through preparation technology, explain that preparation process and product quality can reach stable and controllable basically.
The freezing-thawing test of embodiment 7 Ginkgo Leaf Extract and Dipyridamole Injection
The Ginkgo Leaf Extract and Dipyridamole Injection that embodiment 1～3 is prepared is placed on respectively below-20 ℃ and carries out freezing-thawing test; Investigate appearance character, pH, Ginkgo total flavones content, Semen Ginkgo terpene lactone contents and the dipyridamole content of each Ginkgo Leaf Extract and Dipyridamole Injection respectively, the result is as shown in table 1.
Table 3 Ginkgo Leaf Extract and Dipyridamole Injection freezing-thawing test result
Visible according to table 2: in the preparation process of Ginkgo Leaf Extract and Dipyridamole Injection, no matter the terpene lactone composition of Semen Ginkgo is after organic solvent extraction is refining, to be added in the extracting solution, still in the preparation process, adds; Prepared sample is all more stable, meets each item requirement of injection, and sample is after the freezing thawing again of freezing; Measure each index of correlation; Sample is all more stable, particulate matter do not occur, explains that the sample of four case study on implementation preparations all can reach the correlated quality requirement.
The stability experiment research of embodiment 8 Ginkgo Leaf Extract and Dipyridamole Injection
1. accelerated test
This test objective is through quickening the chemistry or the physical change of medicine, inquiring into stability of drug, for drug packaging, transportation and storage provide necessary data.Experimental technique: the Ginkgo Leaf Extract and Dipyridamole Injection sample of getting embodiment 1～4 preparation; In 40 ℃ ± 2 ℃, relative humidity is 75% ± 5% condition held 6 months; Take a sample respectively 0,1,2,3 and 6 the end of month, the content of outward appearance, particulate matter, pH value, Semen Ginkgo terpene lactone, total flavones and dipyridamole is investigated.Result of the test sees the following form.
Table 4 accelerated test result
According to result of the test, the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation was packaged into 10ml/ by the commercially available prod specification and props up, the condition held of 40 ± 2 ℃ of temperature, relative humidity 75 ± 5% 6 months.Through sample analysis, analysis result and comparison in 0 month, the content of Semen Ginkgo terpene lactone, total flavones and dipyridamole descends a little, and other each items detect indexs does not have significant change, and quality is basicly stable.
2. influence factor's test
2.1 hot test
Get the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation; In temperature is to place 20 days in 60 ℃ the thermostatic drying chamber; In the 0th day, the 5th day, the 10th day and sampling in the 20th day; Observe the outward appearance of Ginkgo Leaf Extract and Dipyridamole Injection and the situation of change of particulate matter, measure the content of each sample pH value, Semen Ginkgo terpene lactone, total flavones and dipyridamole.The result sees table 5.
Table 5 hot test result
According to result of the test, the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation was packaged into 10ml/ by the commercially available prod specification and props up, the condition held of 60 ± 2 ℃ of temperature 20 days.Through sample analysis, analysis result and comparison in 0 day, the content of Semen Ginkgo terpene lactone, total flavones and dipyridamole descends a little, and other each items detect indexs does not have significant change, and quality is basicly stable.
2.2 strong illumination test
Get the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation; In illumination is the condition held 20 days of 4500lx ± 5001x; In the 0th day, the 5th day, the 10th day and sampling in the 20th day; Observe the outward appearance of Ginkgo Leaf Extract and Dipyridamole Injection and the situation of change of particulate matter, the content of the pH value of working sample, Semen Ginkgo terpene lactone, total flavones and dipyridamole.The result sees table 6.
Table 6 strong illumination result of the test
According to result of the test, the Ginkgo Leaf Extract and Dipyridamole Injection sample of embodiment 1～4 preparation is packaged into 10ml/ by the commercially available prod specification and props up, and is the condition held 20 days of 4500lx ± 500lx in illumination.Through sample analysis, analysis result and comparison in 0 day, the content of Semen Ginkgo terpene lactone, total flavones and dipyridamole descends obviously, and other each items detect indexs does not have significant change.Interpret sample preferably keeps in Dark Place.
Claims (6)
1. the method for preparing of a Ginkgo Leaf Extract and Dipyridamole Injection, it comprises the steps:
1) collection of Folium Ginkgo, processing, detection and classification;
2) from Folium Ginkgo, extract Folium Ginkgo terpene lactone extract;
3) extract the Folium Ginkgo flavone extract;
4) will contain Semen Ginkgo total flavones 10g the Folium Ginkgo flavone extract and 2 the gram dipyridamoles add water backflow after-filtration, process mixed liquor 1;
5) above-mentioned full dose mixed liquor 1, the Folium Ginkgo terpene lactone extract that contains Ginkgo total lactones 5g, 2 gram dipyridamoles and adjuvant are mixed; Process Ginkgo Leaf Extract and Dipyridamole Injection by the conventional formulation method; In the said Ginkgo Leaf Extract and Dipyridamole Injection; The content of Ginkgo total flavones and dipyridamole is except that should meeting conventional the requirement, and the content of Semen Ginkgo terpene lactone should be 0.5～1.2mg/mL.
2. method for preparing according to claim 1; It is characterized in that; Step 1) is specially: be that late June is to late October the collection period of Folium Ginkgo; Bright leaf at source in carrying out drying below 60 ℃, is selected for use the total content of ginkalide A, ginkalide B, ginkalide C and bilobalide to be higher than 0.35% Folium Ginkgo and extracted.
3. method for preparing according to claim 1 is characterized in that step 2) be specially: Folium Ginkgo is put in the extraction element, added 40～80% organic solvent of 7～15 times of amount medical material weight; Heating and refluxing extraction, wherein, organic solvent is selected any one in methanol, ethanol, acetone, the acetonitrile for use, extracts 1～3 hour; Extract merge extractive liquid,, decompression and solvent recovery under the low temperature altogether 1～3 time; Concentrate drying makes the organic solvent crude extract of Folium Ginkgo, stays the Folium Ginkgo medicinal residues after the extraction; Crude extract adds ethyl acetate, extracts after-filtration, obtain filtrating with ethyl acetate extraction after crude extract 2; Merging filtrate, decompression filtrating is reclaimed ethyl acetate, adds 80～100% alcoholic solution, makes abundant dissolving, filters; In the polyamide chromatographic column of having handled well, carry out eluting on the filtrating, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with 80～100% alcoholic solution; In alumina column, carry out eluting on the concentrated solution, collect the effluent and the eluent of bilobalide-containing, concentrating under reduced pressure with dehydrated alcohol; Crystallization is filtered, and drying promptly gets Folium Ginkgo terpene lactone extract.
4. method for preparing according to claim 1 is characterized in that step 3) is specially: with Folium Ginkgo medicinal residues behind the organic solvent extraction and the crude extract 2 behind the ethyl acetate extraction; Add 7～15 times purified water of medicinal residues and crude extract 2 gross weights, heating and refluxing extraction was extracted 1～3 hour; Extract altogether 1～3 time, merge extractive liquid, is evaporated to 1/16～1/8 of original volume; With concentrated solution placement, deposition, centrifugal, filtration, filtrating going up adsorbed in the macroporous adsorptive resins of having handled well, wherein; Can select any among macroporous adsorbent resin AB-8, ADS-17, D101, DM130, D201, HPD417, HPD600, HPD722, HZ803, HZ806, the HZ841 for use, extracting solution washs with the purified water of 8～12 times of column volume amounts after abundant absorption; Use ethanol elution then, collect the apricot yellow ketone part of argentiferous eluent, be concentrated into and do not have the alcohol flavor, add the purified water of 0.5～2 times of Folium Ginkgo weight then; Stir leave standstill after the cooling, deposition, centrifugal, filter, will filtrate concentrates receipts cream, is dried to water content below 6%; Pulverize, promptly get the Semen Ginkgo water extract; The concentration expressed in percentage by volume that adds 8～12 times of weight in the Folium Ginkgo water extract is the ethanol more than 90%, is stirred to dissolving, filters; Filtrating is concentrated into nothing alcohol flavor, adds 10 times of purified water that concentrate the back medicine liquid volumes then, leaves standstill after the stirring cooling; Get supernatant, after centrifugalize, concentrate and receive cream, vacuum drying to water content is below 6%; Pulverize, promptly get the Folium Ginkgo flavone extract.
5. method for preparing according to claim 1 is characterized in that step 4) is specially: get the purified water that the Folium Ginkgo flavone extract and 2 that contains Semen Ginkgo total flavones 10g restrains dipyridamoles and 10～15 times of weight of extract weight, fully stir; Mix homogeneously, adjust pH to 3.5～5.5, reflux 0.5～1 hour; Be cooled to room temperature, supernatant is got in readjustment pH value to 3.5～5.5; With the membrane filtration of 0.2mm, filtrating is the ultrafilter membrane ultrafiltration below 100000 dalton again, the filtrating autoclave sterilization; Put coldly, promptly get and process mixed liquor 1, low tempertaure storage.
6. method for preparing according to claim 1 is characterized in that step 5) is specially: with above-mentioned full dose mixed liquor 1, the Folium Ginkgo terpene lactone extract that contains Ginkgo total lactones 5g, 2 gram dipyridamole and relevant auxiliary materials, dissolve fully with water for injection; Wherein, adjuvant is one or more in propylene glycol, glycerol, Tween 80, sodium pyrosulfite, vitamin C, calcium disodium edetate or the PEG400, and adjust pH is 3.5～5.5, and deepfreeze is placed; Filter, depyrogenation makes the injection intermediate; Through after the assay was approved, use the 0.22mm filtering with microporous membrane, the filtrating fill; Autoclave sterilization promptly gets Ginkgo Leaf Extract and Dipyridamole Injection, low tempertaure storage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210178017 CN102670670B (en) | 2012-06-01 | 2012-06-01 | Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201210178017 CN102670670B (en) | 2012-06-01 | 2012-06-01 | Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102670670A true CN102670670A (en) | 2012-09-19 |
| CN102670670B CN102670670B (en) | 2013-10-16 |
Family
ID=46803656
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201210178017 Active CN102670670B (en) | 2012-06-01 | 2012-06-01 | Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102670670B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104800218A (en) * | 2014-01-27 | 2015-07-29 | 贵州益佰制药股份有限公司 | Ginkgo dipyridolum composition, preparation thereof, preparation method of composition, and preparation method of preparation |
| CN105412167A (en) * | 2015-11-27 | 2016-03-23 | 陕西嘉禾生物科技股份有限公司 | Method for extracting and separating ginkgo flavone and ginkgolide from ginkgo leaves |
| CN107184616A (en) * | 2017-05-07 | 2017-09-22 | 珠海亿邦医药有限公司 | A kind of preparation method of Ginkgo Damo injection |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101628003A (en) * | 2009-08-19 | 2010-01-20 | 上海杏灵科技药业股份有限公司 | Extract of ginkgo biloba leaves and dipyridamole composite and preparation method and application thereof |
-
2012
- 2012-06-01 CN CN 201210178017 patent/CN102670670B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101628003A (en) * | 2009-08-19 | 2010-01-20 | 上海杏灵科技药业股份有限公司 | Extract of ginkgo biloba leaves and dipyridamole composite and preparation method and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104800218A (en) * | 2014-01-27 | 2015-07-29 | 贵州益佰制药股份有限公司 | Ginkgo dipyridolum composition, preparation thereof, preparation method of composition, and preparation method of preparation |
| CN105412167A (en) * | 2015-11-27 | 2016-03-23 | 陕西嘉禾生物科技股份有限公司 | Method for extracting and separating ginkgo flavone and ginkgolide from ginkgo leaves |
| CN105412167B (en) * | 2015-11-27 | 2020-01-21 | 陕西嘉禾生物科技股份有限公司 | Method for extracting and separating ginkgetin and bilobalide from folium Ginkgo |
| CN107184616A (en) * | 2017-05-07 | 2017-09-22 | 珠海亿邦医药有限公司 | A kind of preparation method of Ginkgo Damo injection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102670670B (en) | 2013-10-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5940212B2 (en) | Ginkgolide extraction and separation method | |
| CN105294623B (en) | A sesquiterpene lactone compound, its preparation method and application | |
| CN101863871A (en) | Total glycosides of Rhodiola rosea, medical application and preparation method thereof | |
| CN101502585B (en) | Chinese red pepper plant extract as well as preparation method and application thereof | |
| CN107854507B (en) | A kind of method for extracting flavonoids from mugwort leaves | |
| CN103599144B (en) | The preparation method of jatamans valeriana rhizome epoxy iridoid ester active component | |
| CN102058814B (en) | General flavone extractive of four medicaments including grassleaf sweetflag rhizome and preparation method thereof | |
| CN102670670B (en) | Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones | |
| JP2010528063A (en) | Method and use for obtaining an extract containing sequoyitol from a plant belonging to the genus Rhododendron, soybean, genus Ginkgo | |
| CN101756965A (en) | Application of maca imidazole alkaloid in preparation of cardiovascular drugs | |
| CN102846704B (en) | A Leonurus japonicus injection, its preparation method, and method for detecting total alkaloids | |
| CN109053756B (en) | Phenylpropanoid ester type catechin and preparation method and application thereof | |
| CN111948305A (en) | Quality control method and preparation method for producing Qingda granules and compound traditional Chinese medicine based on Qingda granules | |
| WO2014000119A1 (en) | Anti-tachyarrhythmia formulation and preparation method thereof | |
| CN103091412B (en) | Method for detecting ginkgolide effective parts | |
| CN103239487A (en) | Bilobalide injection and content determination method | |
| WO2019113861A1 (en) | Ginkgo biloba extract medicinal raw material and preparation method therefor | |
| CN105535050B (en) | A kind of Radix Angelicae Sinensis antineoplastic | |
| CN103163252B (en) | Determination method of total ginkgolic acid in ginkgolide composition | |
| CN112300185A (en) | Hepatotoxicity-reduced alkaloid compound, preparation method and use thereof | |
| CN102423326B (en) | The use of the total flavonoids of bell flower and primrose | |
| CN106420850B (en) | Compositions extracted from gingko biloba leaves and its preparing the application in Shu Xuening injection | |
| CN103239486B (en) | Residue determination method of ginkgo lactone composition for treating cardiovascular and cerebrovascular diseases | |
| CN1772008B (en) | Preparation method and quality control method of ginkgo damole for injection | |
| CN108159109A (en) | Radix Astragali flavone extract is preparing the application in treating Neuroleptic Leukocytopenia disease drug |
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 | ||
| C56 | Change in the name or address of the patentee | ||
| CP03 | Change of name, title or address |
Address after: 037010 the first medical zone of Datong Economic and Technological Development Zone, Shanxi Patentee after: Shanxi Powerdone Pharmaceutics Co., Ltd. Address before: 037010 Datong economic and Technological Development Zone, Shanxi Patentee before: Shanxi Powerdone Pharmaceutical Co., Ltd. |