CN108362809B - Quality evaluation method of ginkgo leaf and extract and preparation thereof - Google Patents
Quality evaluation method of ginkgo leaf and extract and preparation thereof Download PDFInfo
- Publication number
- CN108362809B CN108362809B CN201810103484.0A CN201810103484A CN108362809B CN 108362809 B CN108362809 B CN 108362809B CN 201810103484 A CN201810103484 A CN 201810103484A CN 108362809 B CN108362809 B CN 108362809B
- Authority
- CN
- China
- Prior art keywords
- glucoside
- reference substance
- kaempferol
- ginkgo
- rhamnoside
- 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 - Fee Related
Links
- 241000218628 Ginkgo Species 0.000 title claims abstract description 40
- 235000011201 Ginkgo Nutrition 0.000 title claims abstract description 40
- 235000008100 Ginkgo biloba Nutrition 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 239000000284 extract Substances 0.000 title claims abstract description 20
- 238000013441 quality evaluation Methods 0.000 title claims abstract description 8
- 239000013558 reference substance Substances 0.000 claims abstract description 61
- 238000012937 correction Methods 0.000 claims abstract description 21
- 239000000126 substance Substances 0.000 claims abstract description 14
- 238000011156 evaluation Methods 0.000 claims abstract description 13
- 229930003935 flavonoid Natural products 0.000 claims abstract description 13
- 150000002215 flavonoids Chemical class 0.000 claims abstract description 13
- 235000017173 flavonoids Nutrition 0.000 claims abstract description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 42
- 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
- 229930003944 flavone Natural products 0.000 claims description 24
- 235000011949 flavones Nutrition 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
- 150000002212 flavone derivatives Chemical class 0.000 claims description 23
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 20
- 239000011550 stock solution Substances 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 13
- 238000005303 weighing Methods 0.000 claims description 12
- 238000004364 calculation method Methods 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- 239000012488 sample solution Substances 0.000 claims description 9
- 239000012085 test solution Substances 0.000 claims description 9
- FVQOMEDMFUMIMO-UHFFFAOYSA-N Hyperosid Natural products OC1C(O)C(O)C(CO)OC1OC1C(=O)C2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 FVQOMEDMFUMIMO-UHFFFAOYSA-N 0.000 claims description 7
- QJTYCCFDQWFJHU-UHFFFAOYSA-N Quercetin-5-O-beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC(O)=CC2=C1C(=O)C(O)=C(C=1C=C(O)C(O)=CC=1)O2 QJTYCCFDQWFJHU-UHFFFAOYSA-N 0.000 claims description 7
- 150000001875 compounds Chemical class 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- GXMWXESSGGEWEM-UHFFFAOYSA-N isoquercitrin Natural products OCC(O)C1OC(OC2C(Oc3cc(O)cc(O)c3C2=O)c4ccc(O)c(O)c4)C(O)C1O GXMWXESSGGEWEM-UHFFFAOYSA-N 0.000 claims description 7
- OVSQVDMCBVZWGM-QSOFNFLRSA-N quercetin 3-O-beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O OVSQVDMCBVZWGM-QSOFNFLRSA-N 0.000 claims description 7
- SZDMSNWMQAMVTJ-UHFFFAOYSA-N quercetin-3-O-glucoside Natural products OC1OC(COC2=C(C(=O)c3cc(O)cc(O)c3O2)c4ccc(O)c(O)c4)C(O)C(O)C1O SZDMSNWMQAMVTJ-UHFFFAOYSA-N 0.000 claims description 7
- FZKBNCDAGYDHTP-UHFFFAOYSA-N quercetin-3-O-glycoside Natural products OC1C(O)C(O)C(O)OC1COC1=C(C=2C=C(O)C(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O FZKBNCDAGYDHTP-UHFFFAOYSA-N 0.000 claims description 7
- 238000007865 diluting Methods 0.000 claims description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 6
- 238000005516 engineering process Methods 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- SPUFXPFDJYNCFD-YQJBXTIASA-N 2-(3,4-dihydroxyphenyl)-5-hydroxy-7-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](OC=2C(C3=C(O)C=C(O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 SPUFXPFDJYNCFD-YQJBXTIASA-N 0.000 claims description 4
- 239000012982 microporous membrane Substances 0.000 claims description 4
- SPUFXPFDJYNCFD-UHFFFAOYSA-N quercetin 3-O-rutinoside-7-O-glucoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(OC4C(C(O)C(O)C(CO)O4)O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 SPUFXPFDJYNCFD-UHFFFAOYSA-N 0.000 claims description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 3
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 claims description 3
- 239000009429 Ginkgo biloba extract Substances 0.000 claims description 3
- NRSUOHVIKLCXEE-UHFFFAOYSA-N Isorhamnetin 3-O-beta-D-glucopyranoside Natural products CCC1OC(OC2=C(Oc3cc(O)cc(O)c3C2=O)c4ccc(O)c(OC)c4)C(O)C(O)C1O NRSUOHVIKLCXEE-UHFFFAOYSA-N 0.000 claims description 3
- RTATXGUCZHCSNG-TYSPDFDMSA-N Kaempferol-3-O-rutinoside Natural products OC1[C@H](O)[C@@H](O)C(C)O[C@H]1OCC1[C@@H](O)[C@@H](O)C(O)[C@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=CC(O)=CC=2)=O)O1 RTATXGUCZHCSNG-TYSPDFDMSA-N 0.000 claims description 3
- RTATXGUCZHCSNG-QHWHWDPRSA-N Nicotiflorin Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=CC(O)=CC=2)=O)O1 RTATXGUCZHCSNG-QHWHWDPRSA-N 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 235000019253 formic acid Nutrition 0.000 claims description 3
- CQLRUIIRRZYHHS-LFXZADKFSA-N isorhamnetin 3-O-beta-D-glucopyranoside Chemical compound C1=C(O)C(OC)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)=C1 CQLRUIIRRZYHHS-LFXZADKFSA-N 0.000 claims description 3
- RTATXGUCZHCSNG-ZFDPGQBLSA-N nicotiflorin Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](OC2=C(c3ccc(O)cc3)Oc3c(c(O)cc(O)c3)C2=O)O1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1 RTATXGUCZHCSNG-ZFDPGQBLSA-N 0.000 claims description 3
- RTATXGUCZHCSNG-UHFFFAOYSA-N nicotiflorine Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=CC(O)=CC=2)=O)O1 RTATXGUCZHCSNG-UHFFFAOYSA-N 0.000 claims description 3
- MQVRGDZCYDEQML-UHFFFAOYSA-N Astragalin Natural products C1=CC(OC)=CC=C1C1=C(OC2C(C(O)C(O)C(CO)O2)O)C(=O)C2=C(O)C=C(O)C=C2O1 MQVRGDZCYDEQML-UHFFFAOYSA-N 0.000 claims 3
- HOUHSBKVSRPPGO-UHFFFAOYSA-N UNPD177615 Natural products OCC(O)C1OC(OC2=C(Oc3cc(O)cc(O)c3C2=O)c4ccc(O)cc4)C(O)C1O HOUHSBKVSRPPGO-UHFFFAOYSA-N 0.000 claims 3
- JPUKWEQWGBDDQB-QSOFNFLRSA-N kaempferol 3-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C(C=2C=CC(O)=CC=2)OC2=CC(O)=CC(O)=C2C1=O JPUKWEQWGBDDQB-QSOFNFLRSA-N 0.000 claims 3
- ZZZILDYSXRHUNY-UHFFFAOYSA-N kaempferol-3-O-glucoside Natural products OC1OC(COC2=C(Oc3cc(O)cc(O)c3C2=O)c4ccc(O)cc4)C(O)C(O)C1O ZZZILDYSXRHUNY-UHFFFAOYSA-N 0.000 claims 3
- 238000009210 therapy by ultrasound Methods 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 11
- 239000000463 material Substances 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 239000007791 liquid phase Substances 0.000 abstract description 3
- 238000003908 quality control method Methods 0.000 abstract description 3
- 239000012071 phase Substances 0.000 description 20
- IYRMWMYZSQPJKC-UHFFFAOYSA-N kaempferol Chemical compound C1=CC(O)=CC=C1C1=C(O)C(=O)C2=C(O)C=C(O)C=C2O1 IYRMWMYZSQPJKC-UHFFFAOYSA-N 0.000 description 16
- 239000007788 liquid Substances 0.000 description 16
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 14
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 description 12
- 230000014759 maintenance of location Effects 0.000 description 11
- UBSCDKPKWHYZNX-UHFFFAOYSA-N Demethoxycapillarisin Natural products C1=CC(O)=CC=C1OC1=CC(=O)C2=C(O)C=C(O)C=C2O1 UBSCDKPKWHYZNX-UHFFFAOYSA-N 0.000 description 8
- 235000008777 kaempferol Nutrition 0.000 description 8
- UXOUKMQIEVGVLY-UHFFFAOYSA-N morin Natural products OC1=CC(O)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 UXOUKMQIEVGVLY-UHFFFAOYSA-N 0.000 description 8
- 239000012086 standard solution Substances 0.000 description 7
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 6
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000011002 quantification Methods 0.000 description 6
- 235000005875 quercetin Nutrition 0.000 description 6
- 229960001285 quercetin Drugs 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 5
- 210000000582 semen Anatomy 0.000 description 5
- 229930182470 glycoside Natural products 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- SQGLUEWZRKIEGS-UHFFFAOYSA-N Ginkgetin Natural products C1=CC(OC)=CC=C1C1=CC(=O)C2=C(O)C=C(OC)C(C=3C(=CC=C(C=3)C=3OC4=CC(O)=CC(O)=C4C(=O)C=3)O)=C2O1 SQGLUEWZRKIEGS-UHFFFAOYSA-N 0.000 description 3
- 229930045534 Me ester-Cyclohexaneundecanoic acid Natural products 0.000 description 3
- 238000010812 external standard method Methods 0.000 description 3
- AIFCFBUSLAEIBR-UHFFFAOYSA-N ginkgetin Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C(C=1)=CC=C(OC)C=1C1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=C(O)C=C1 AIFCFBUSLAEIBR-UHFFFAOYSA-N 0.000 description 3
- 150000002338 glycosides Chemical class 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- GQODBWLKUWYOFX-UHFFFAOYSA-N Isorhamnetin Natural products C1=C(O)C(C)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 GQODBWLKUWYOFX-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- IZQSVPBOUDKVDZ-UHFFFAOYSA-N isorhamnetin Chemical compound C1=C(O)C(OC)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)O)=C1 IZQSVPBOUDKVDZ-UHFFFAOYSA-N 0.000 description 2
- 235000008800 isorhamnetin Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 2
- 239000012088 reference solution Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- UFJORDZSBNSRQT-UHFFFAOYSA-N 3-(4-oxo-2-phenylchromen-3-yl)-2-phenylchromen-4-one Chemical compound O1C2=CC=CC=C2C(=O)C(C=2C(C3=CC=CC=C3OC=2C=2C=CC=CC=2)=O)=C1C1=CC=CC=C1 UFJORDZSBNSRQT-UHFFFAOYSA-N 0.000 description 1
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 1
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 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
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- -1 flavone glycosides Chemical class 0.000 description 1
- 229940068052 ginkgo biloba extract Drugs 0.000 description 1
- 235000020686 ginkgo biloba extract Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/884—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
A quality evaluation method of ginkgo leaf and an extract and a preparation thereof belongs to the technical field of quality detection of traditional Chinese medicinal materials. The invention realizes the determination of the contents of various flavonoid chemical components in the ginkgo by constructing the high-performance liquid phase fingerprint of the chemical components of the ginkgo and calculating the correction factor by means of a one-test-multiple-evaluation method. The method has simple operation and good stability, can objectively, comprehensively and accurately evaluate the quality of the ginkgo leaves and the extract or preparation thereof, is used for quality control, can solve the problem that the quality of the medicinal materials and the extract or preparation thereof cannot be objectively and reasonably controlled due to the lack of reference substances, and has important significance for controlling the quality and ensuring the curative effect.
Description
Technical Field
The invention relates to the technical field of quality detection of traditional Chinese medicinal materials, in particular to a quality evaluation method of ginkgo leaves and extracts and preparations thereof based on a high performance liquid fingerprint and a one-measurement-multiple-evaluation method.
Technical Field
The folium Ginkgo is Ginkgo of GinkgoaceaeGinkgo bilobaL, the dried leaves have the effects of promoting blood circulation, removing blood stasis, dredging collaterals, relieving pain, astringing lung, relieving asthma, eliminating turbid pathogen, and reducing blood lipid, the ginkgo has rich chemical components including flavone, lactone, phenolic acid, etc., the ginkgo flavone has the most content and various compositions, and the ginkgo extract with the total flavone content of 24 percent can improve various diseases of cardiovascular and cerebrovascular systems, and has no side effect.
At present, in the quality standard of ginkgo leaf of 'Chinese pharmacopoeia' 2015 edition, the content of total flavonoids is evaluated and controlled by measuring the content of quercetin, kaempferol and isorhamnetin after acid hydrolysis, the method is complex in pretreatment, long in time consumption and incapable of fully reflecting the real and accurate content of flavonoid components.
Therefore, the invention provides a quality evaluation method of ginkgo leaves and extracts and preparations thereof based on a high performance liquid fingerprint and a one-measurement-multiple-evaluation method. The method takes one flavone component in ginkgo as an internal reference substance, and realizes the determination of the contents of various flavonoid chemical components in ginkgo by calculating relative correction factors. The method is simple and convenient to operate, has good stability, can objectively, comprehensively and accurately evaluate the quality of the ginkgo leaves and the extract and the preparation thereof, can solve the problem that the quality cannot be objectively and reasonably evaluated due to the lack of reference substances, and has important significance for controlling the quality and ensuring the curative effect.
Disclosure of Invention
The invention aims to provide a content determination method for determining multiple flavone components in ginkgo aiming at the defects of the prior art, which can not only accurately evaluate the quality of ginkgo leaves and extracts thereof and total flavone in preparations, but also solve the problem that the quality of traditional Chinese medicinal materials and preparations thereof cannot be objectively and reasonably controlled due to the lack of reference substances.
In order to achieve the purpose, the invention adopts the following technical scheme:
the method comprises the steps of measuring the content of a plurality of flavonoid chemical components in the ginkgo by utilizing a high performance liquid chromatography technology and a multi-evaluation method, so as to realize objective, comprehensive and accurate quality evaluation of the ginkgo and the extract thereof and the preparation, wherein the content measurement of the flavonoid chemical components refers to the content measurement of the ginkgo and the extract thereof and a plurality of flavonoid reference substances in the preparation, and the number of the flavonoid chemical components is 8-16.
The method specifically comprises the following steps:
1) preparing a reference substance solution: precisely weighing 1-5mg of each flavone reference substance respectively, placing in a volumetric flask, dissolving with 80-100% methanol to constant volume, filtering to obtain stock solutions of each single reference substance, weighing single standard substance solutions with different volumes respectively, mixing and diluting to obtain mixed samples containing multiple flavone components as mixed reference substance solutions for later use;
2) preparing a test solution: precisely weighing folium Ginkgo, semen Ginkgo extract or semen Ginkgo preparation, adding 50-100% methanol, ultrasonic or heating for extraction, filtering, and filtering the filtrate with 0.45um or 0.22um filter membrane to obtain sample solution;
3) calculating a correction factor RCF by taking the mixed reference solution prepared in the step 1) at a concentration of 2-20 μ L, determining chromatogram by high performance liquid chromatography, calculating peak area, selecting one compound as an internal reference, and calculating a relative correction factor f between the internal reference and the other compoundsk/s。
Wherein, the relative correction factor calculation formula is as follows: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesThe peak area of the internal reference substance is shown; csThe concentration of the internal reference substance is shown; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Measuring active ingredients in the sample by taking the sample solution prepared in step 2) 2-20 μ L, measuring chromatogram with high performance liquid chromatography, and calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sTo correct the factor, AsThe peak area of the internal reference substance is shown.
The invention accurately evaluates the content of the total flavone of ginkgo by measuring the total amount of 8-16 flavone glycosides and aglycone thereof with high content and good activity in ginkgo leaves and extracts thereof and preparations.
The flavone control product comprises quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L-D-glucoside, kaempferol-3-0- (2' ', 6' ' -di-O-L2-L-rhamnosyl) -L-D-glucoside, kaempferol-3-O-L sophoroside, quercetin-3-O-rutinoside-7-O-glucoside, 3' -methyl-myricetin-3-O-rutinoside, quercetin 3-O-glucoside, quercetin 3-O- (2-L-D-glucosyl-L-L-rhamnoside), kaempferol-3-O-rutinoside, isorhamnetin-3-O- β -D-glucoside, kaempferol 3- (2' ' - β -D-glucoside) -L-L-rhamnoside, kaempferol-3-O-6-rhamnoside- (2' ' -p-581-p-rhamnoside), and coumarin-3-O-3-O-glucoside, and coumarin-3-O-3-acyl-5968-rhamnoside.
The invention constructs a characteristic liquid phase fingerprint by a high performance liquid chromatography technology and ensures that the flavonoid contrast component has good separation degree (the separation degree is more than or equal to 1.2).
The detection conditions of the high performance liquid chromatography are as follows: the detector of the high performance liquid chromatograph uses a diode array detector or a universal detector, uses octa-or octadecylsilane chemically bonded silica as a filler, acetonitrile as a mobile phase A, and 0.1% formic acid aqueous solution as a mobile phase B, and performs gradient elution, wherein in the gradient elution process, the proportion of the mobile phase A, B is changed as follows: 0-20min, 15-20% of phase A and 85-80% of phase B; 20-25min, 20-30% of phase A and 80-70% of phase B; 25-30min, 30-35% of phase A and 70-65% of phase B; 30-45min, 35-40% of phase A and 65-60% of phase B. The ultraviolet detector is used for detecting the wavelength of 210nm-360 nm. The flow rate is 0.5ml/min-1.5ml/min, and the column temperature is kept at 20-45 ℃.
The high performance liquid phase fingerprint spectrum is obtained by applying the conditions, the similarity of all spectra is not lower than 0.90, and the RSD of each peak area is not higher than 2%.
The high performance liquid chromatography column model is preferably Gemini C18 (250 mm × 4.6.6 mm, 5.0 μm), the flow rate is preferably 0.8m L/min, the detection wavelength is preferably 360nm, and the column temperature is preferably 35 ℃.
The invention adopts a one-test-multiple-evaluation method, takes one component as an internal reference substance, and performs relative calibrationAnd (4) calculating the content of each compound respectively by using a positive factor. Wherein, the relative correction factor calculation formula is as follows: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesThe peak area of the internal reference substance is shown; csThe concentration of the internal reference substance is shown; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
The invention has the following advantages:
1) compared with the traditional method, the invention solves the problems of complicated operation, lack of reference substances and the like in the multi-component quantification and multi-index quality control of the traditional Chinese medicine, realizes the multi-index synchronous quality control while reducing the detection components, and accurately reflects the quality of the traditional Chinese medicine product.
2) The invention combines the high performance liquid chromatography technology with a one-test-and-multiple-evaluation method, double standard-reaching products which meet the requirements of both fingerprint similarity and content are determined as qualified products, and the method has important significance for controlling the quality consistency among different batches of products.
3) The method of the invention takes a flavone component in ginkgo as a reference substance, and simultaneously quantifies the content of a plurality of flavone components such as quercetin and glycosides thereof, kaempferol and glycosides thereof, isorhamnetin and glycosides thereof, ginkgo biflavone and the like in ginkgo by a one-test-multiple-evaluation method, thereby accurately evaluating the content of total flavone in ginkgo leaf and extract thereof and preparations.
4) The one-test-multiple-evaluation method provided by the invention is simple and convenient to operate, and the multi-index quality evaluation method is good in stability, can objectively, comprehensively and accurately evaluate the quality of the ginkgo leaves and the extract and preparation thereof, avoids the evaluation method for measuring the content of flavone component hydrolysate in the traditional method, and can effectively prevent the problems of product counterfeiting, process modification and the like.
Drawings
FIG. 1 is a high performance liquid chromatogram of a Ginkgo biloba extract.
FIG. 2 is a high performance liquid chromatogram of a ginkgo leaf mixed standard consisting of 8 flavonoids.
FIG. 3 is a high performance liquid chromatogram of a ginkgo leaf mixed standard consisting of 10 kinds of flavonoids.
FIG. 4 is a high performance liquid chromatogram of a ginkgo leaf mixed standard consisting of 12 flavonoids.
Detailed Description
The present invention is further illustrated by the following examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure, and which are intended to cover all modifications, equivalents, and equivalents that may be resorted to by those skilled in the art to which the invention pertains.
Chromatographic conditions used in the following examples:
1) the instrument comprises the following steps: high performance liquid chromatography (hua spectrum S6000, dahlianhua spectrum science and technology development limited);
2) analyzing a chromatographic column: gemini C18(250 mm × 4.6.6 mm, 5.0 μm, Philomen, USA);
3) mobile phase: acetonitrile is taken as a mobile phase A, 0.1 percent formic acid aqueous solution is taken as a mobile phase B,
4) gradient conditions: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A
5) Detector conditions: an ultraviolet detector with a detection wavelength of 360nm
6) The flow rate is 0.8 ml/min;
7) the column temperature was 35 ℃.
Example 1
The following detailed description is made with reference to fig. 2 and the embodiments:
1) reference substance
quercetin-3-O- (2'', 6'' -di-O- α -L-rhamnosyl) -L1-D-glucoside (1), kaempferol-3-0- (2'', 6'' -di-O-L0-L-rhamnosyl) -L3-D-glucoside (2), quercetin-3-O-rutinose-7-O-glucoside (3), quercetin 3-O-glucoside (4), isorhamnetin-3-O- β -D-rutinoside (5), kaempferol 3- (2'' - β -D-glucoside) -L2-L-rhamnoside (6), quercetin 3-O- α - (6'' '-p-coumaroyl-glucoside- β -1, 4-rhamnoside) (7), kaempferol 3-O- α - (6' '' -p-coumaroyl-glucoside- β -1, 2-rhamnoside) (8).
2) Preparation of sample solution
Precisely weighing semen Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, ultrasonic treating for 5min, diluting to constant volume of 20m L, shaking, filtering with 0.22 μm microporous membrane to obtain test solution, and analyzing and detecting by high performance liquid chromatography.
3) Preparation of control solutions
Precisely weighing 1.0mg of each of the 8 ginkgetin monomer compounds in the step 1), respectively adding methanol into volumetric flasks to dissolve and fix the volume to 2ml, shaking up to prepare stock solutions of all reference substances, wherein the concentration is 0.5mg/m L, precisely measuring the stock solutions of all reference substances to be 2.0m L, placing the stock solutions into the same volumetric flask with the volume of 25m L, adding methanol to dissolve and dilute to a scale, shaking up to obtain a reference substance mixed solution, and feeding the reference substance mixed solution into a high-performance liquid chromatograph.
4) Calculation of correction factors
Taking 10 mu L of the mixed reference substance solution prepared in the step 3), measuring chromatogram by using high performance liquid chromatography, calculating peak area, selecting No. 4 reference substance as an internal reference substance, and respectively calculating relative correction factor f between the internal reference substance and other compoundsk/s. The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sIs a correction factor，AsThe peak area of the internal reference substance is shown.
The gradient condition is as follows: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A;
example 2
The following detailed description is made with reference to fig. 3 and the embodiments:
1) reference substance
quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L1-D-glucoside (1), kaempferol-3-0- (2' ', 6' ' -di-O-L0-L-rhamnosyl) -L3-D-glucoside (2), quercetin-3-O-rutinoside-7-O-glucoside (3), 3' -methyl-myricetin-3-O-rutinoside (4), quercetin 3-O-glucoside (5), kaempferol-3-O-rutinoside (6), isorhamnetin-3-O- β -D-rutinoside (7), kaempferol 3- (2' ' - β -D-glucoside) -L2-L-rhamnoside (8), quercetin 3-O- α - (6' ' ' -p-coumaroyl-glucoside- β -1, 4-rhamnoside) (9), kaempferol 3-O- α - (6' ' ' -p-coumaroyl-glucoside- β -1, 4-rhamnoside) (9 ' ' -rhamnosyl- α -p-O-3-O-rhamnoside) (β).
2) Preparation of sample solution
Precisely weighing semen Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, ultrasonic treating for 5min, diluting to constant volume of 20m L, shaking, filtering with 0.22 μm microporous membrane to obtain test solution, and analyzing and detecting by high performance liquid chromatography.
3) Preparation of control solutions
Precisely weighing 1.0mg of each of the 10 ginkgetin monomer compounds in the step 1), respectively adding methanol into volumetric flasks to dissolve and fix the volume to 2.0ml, shaking up to prepare stock solutions of each reference substance, wherein the concentration is 0.5mg/m L, precisely measuring the stock solutions of each reference substance by 2.0m L, placing the stock solutions into the same volumetric flask with the volume of 25m L, adding methanol to dissolve and dilute to a scale, shaking up to obtain a reference substance mixed solution, and feeding a filter membrane into a high performance liquid chromatograph.
4) Calculation of correction factors
Collecting the mixed control solution 10 μ L prepared in step 3), and measuring chromatogram with high performance liquid chromatographyCalculating peak area, selecting No. 5 reference as internal reference, and calculating relative correction factor f between the internal reference and other compoundsk/s. The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sTo correct the factor, AsThe peak area of the internal reference substance is shown.
The gradient condition is as follows: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A;
example 3
The following detailed description is made with reference to fig. 4 and the embodiments:
1) reference substance
quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L-D-glucoside (1), kaempferol-3-0- (2' ', 6' ' -di-O-L2-L-3-rhamnosyl) -L-D-glucoside (2), kaempferol-3-O-L5 sophoroside (3), quercetin-3-O-rutinoside-7-O-glucoside (4), 3' -methyl-myricetin-3-O-rutinoside (5), quercetin 3-O-glucoside (6), quercetin 3-O- (2-L-D-glucosyl-L-L-rhamnoside) (7), kaempferol-3 ' ' -O-rutinoside (8), isorhamnetin-3-O- β -D-glucoside (9), kaempferol 3- (2' ' - β -D-6-D-rutinoside) -3985-p-586-rhamnoside (3-O- α), and kaempferol-3-O-636-glucoside (3-O-2 ' ' -6-rhamnoside) (3-2 ' ' -366), and kaempferol-3-O-3-2 ' ' -6-O-glucoside (3-4-L).
2) Preparation of sample solution
Precisely weighing semen Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, ultrasonic treating for 5min, diluting to constant volume of 20m L, shaking, filtering with 0.22 μm microporous membrane to obtain test solution, and analyzing and detecting by high performance liquid chromatography.
3) Preparation of control solutions
Precisely weighing 1.0mg of each of the 12 ginkgetin monomer compounds in the step 1), respectively adding methanol into volumetric flasks to dissolve and fix the volume to 2ml, shaking up to prepare stock solutions of all reference substances, wherein the concentration is 0.5mg/m L, precisely measuring 1.5m L of the stock solutions of all reference substances, placing the stock solutions in the same volumetric flask with the volume of 25m L, adding methanol to dissolve and dilute to a scale, shaking up to obtain a reference substance mixed solution, and feeding the reference substance mixed solution into a high-performance liquid chromatograph.
4) Calculation of correction factors
Taking 10 mu L of the mixed reference substance solution prepared in the step 3), measuring chromatogram by using high performance liquid chromatography, calculating peak area, selecting No. 6 reference substance as an internal reference substance, and respectively calculating relative correction factor f between the internal reference substance and other compoundsk/s. The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk). In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckIs the concentration of a component k to be measured.
4) Determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkAs a group to be testedPeak area of fraction fk/sTo correct the factor, AsThe peak area of the internal reference substance is shown.
The gradient condition is as follows: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A;
example 4
The invention will be further explained below in connection with fig. 3
Methodological investigation example
1) Linear relationship, minimum detection limit (L OD) and minimum quantitation limit (L OQ) assays
Precisely sucking standard solution 8, 20, 40, 80, 160, 320 and 640ug/ml of 10 compounds serving as reference substances, feeding the standard solution into a liquid chromatograph with the sample amount of 10 u L, recording peak areas, drawing a standard curve by taking the concentration of each reference substance as a horizontal coordinate and the peak area as a vertical coordinate, determining the detection limit of the method by S/N =3, determining the quantification limit of the method by S/N =10, determining parameters such as the quantification limit, the detection limit, the quantification limit and a linear regression equation, and displaying a good linear relation (r) by the result2Greater than or equal to 0.9993), detection limit of 0.13-1.11, quantification limit of 0.43-3.69, RSD of relative correction factor of 0.23-1.34, and relative retention time of 0.05-0.72.
Regression equation, linear range, and minimum detection limit and minimum quantification limit
2) Precision, repeatability, stability and accuracy tests:
precision: taking the reference substance solution, repeatedly injecting samples for 6 times in one day and 6 times in 3 consecutive days under the determined chromatographic conditions, and evaluating the precision in the day and the day by the Relative Standard Deviation (RSD) and Retention Time (RT) of the peak areas of the analyzed components, wherein the precision result shows that the RSD of the A (peak area) and the RT (retention time) in the day is respectively lower than 0.89 and 0.75, and the RSD of the A (peak area) and the RT (retention time) in the day is respectively lower than 1.86 and 0.6, thereby indicating that the precision is good.
And (3) stability test: and (3) taking the same mixed standard solution, injecting the mixed standard solution into a liquid chromatograph at 0, 2, 4, 6, 8, 12, 24 and 48 hours respectively, and observing the stability of the mixed standard solution by using the peak area RSD and the Retention Time (RT) of each component, wherein the RSD of the stability A (peak area) and the RSD of the RT (retention time) are respectively 0.66-1.92 and 0.13-1.18, which indicates that the sample solution is stable within 48 hours.
And (3) repeatability test: taking the mixed standard solution (6 parts in parallel), carrying out sample injection analysis under the chromatographic conditions, and evaluating the stability of the mixed standard solution according to the RSD Retention Time (RT) value of each component content in the sample, wherein the RSD of the repeatability A (peak area) and the RSD of the RT (retention time) are respectively 0.67-1.96 and 0.05-0.51, which shows that the repeatability of the sample solution is good.
Precision, repeatability and stability test
Accuracy experiment: through the accuracy of the sample adding recovery rate experimental evaluation method, 1g of a proper amount of ginkgo medicinal material powder with known content is precisely weighed, a certain amount of reference solution is added, samples are prepared according to the sample solution, the samples are prepared into low, medium and high concentrations (50%, 80% and 100%), the sample adding recovery rate is calculated, the recovery rate = (measured value-original value)/the added standard value is 98.8-102.1, the RSD is 0.32-1.76, and the result shows that the method has good accuracy.
Applicability examination of the method:
in order to detect the new method, the laboratory has examined the different liquid chromatographs, the different chromatographic columns and other influencing factors (wavelength, flow rate, column temperature, gradient of mobile phase, gradient time program), the obtained relative correction factor (RPF) and the relative retention time (RSD).
Accurate positioning of chromatographic peaks is a precondition for ensuring application of a one-test-multiple-evaluation method, and positioning is carried out by combining relative retention time value (RRT) parameters with integral features of a chromatogram. RRT = RTx/RTs，RTxRetention time of other chromatographic peaks RTs: retention time of chromatographic peak of internal standard substance, RSD range of RCF of different liquid chromatographs0.12-1.18, and the RSD of RRT is in the range of 0.2-1.36, so that different liquid chromatographs have little influence on the method. The RSD of RCF of different wavelength is in the range of 0.26-2.03, and the RSD of RRT is in the range of 0.53-1.83. The RSD of RCF of different flow rate is in the range of 0.53-1.72, the RSD of RRT is in the range of 0.33-1.65, the RSD of RCF of different column temperature is in the range of 0.65-2.02, the RSD of RRT is in the range of 0.50-1.67, the RCF of mobile phase acid is in the range of 0.65-1.77, the RRT is in the range of 0.46-2.30, and the method is slightly influenced.
Comparison of one-test-multiple evaluation method with external standard method (mg/g)
Note: peak 5 is quercetin 3-O-glucoside
The correlation between the method and the result obtained by the external standard method is represented by the Pearson correlation coefficient, so that the method and the result obtained by the external standard method have no obvious difference.
Claims (1)
1. A quality evaluation method of a ginkgo leaf extract is characterized in that the method is used for measuring the content of a plurality of flavonoid chemical components in the ginkgo leaf extract by utilizing a high performance liquid chromatography technology and a one-measurement-multiple-evaluation method, and comprises the following steps:
1) reference substance
quercetin-3-O- (2' ', 6' ' -di-O- α -L-rhamnosyl) -L-D-glucoside (1), kaempferol-3-O- (2' ', 6' ' -di-O-L2-L-rhamnosyl) -L-D-glucoside (2), kaempferol-3-O-L-sophoroside (3), quercetin-3-O-rutinoside-7-O-glucoside (4), 3' -methyl-myricetin-3-O-rutinoside (5), quercetin-3-O-glucoside (6), quercetin-3-O- (2-L-D-glucosyl-L-L-rhamnoside) (7), kaempferol-3-O-rutinoside (8), isorhamnetin-3-O- β -D-glucoside (9), kaempferol-3- (2' ' -34-D-rhamnoside) -736-rhamnosyl) -L-D-glucoside (1), kaempferol-3-O-glucoside (3-O-636), kaempferol-3-O-glucoside (3-O- α), kaempferol-3-O-glucoside (3- α), kaempferol-3-4-rhamnoside (3-p-366- α, 3-rhamnoside) (3611-rhamnoside-p-rhamnoside) (3-3611- α);
2) preparation of test solution
Precisely weighing folium Ginkgo extract powder 5.0mg in volumetric flask, adding methanol for dissolving, performing ultrasonic treatment for 5min, diluting to constant volume of 20m L, shaking, and filtering with 0.22 μm microporous membrane to obtain sample solution;
3) preparation of Mixed control solutions
Precisely weighing 1.0mg of each of the 12 reference substances in the step 1), respectively placing the reference substances in volumetric flasks, adding methanol for dissolving, fixing the volume to 2ml, shaking up to prepare stock solutions of the reference substances, wherein the concentration of the stock solutions is 0.5mg/m L, precisely weighing 1.5m L of the stock solutions of the reference substances, placing the stock solutions in the same volumetric flask with the volume of 25m L, adding methanol for dissolving, diluting to the scale, and shaking up to obtain mixed reference substance solutions;
4) calculation of correction factors
Taking 10 mu L of the mixed reference substance solution prepared in the step 3), measuring chromatogram by using high performance liquid chromatography, calculating peak area, selecting the No. 6 reference substance quercetin-3-O-glucoside as an internal reference substance, and calculating relative correction factor f between the internal reference substance and other compounds respectivelyk/s；
The relative correction factor calculation formula is: f. ofk/s＝fk/fs＝AkCs/(AsCk) (ii) a In the formula: a. thesPeak area of the internal standard substance; csIs the internal standard concentration; a. thekThe peak area of a certain component k to be measured; ckThe concentration of a certain component k to be measured;
5) determination of other flavone Components
Taking 10 mu L of the test solution prepared in the step 2), measuring a chromatogram by using a high performance liquid chromatography, and respectively calculating the mass of other flavone components according to the following formula;
the formula is: wk=（Ws╳Ak)/(fk/s╳As），WkAs mass of the component to be measured, WsIs the mass of the internal reference substance, AkPeak area of the measured component, fk/sTo correct the factor, AsIs the peak area of the internal reference substance;
the high performance liquid chromatography adopts an analytical chromatographic column: gemini C18；
Mobile phase: acetonitrile is taken as a mobile phase A, and 0.1% formic acid aqueous solution is taken as a mobile phase B;
mobile phase gradient conditions: the ratio of mobile phase A, B varies as: 0-23min, 16-17% A, 23-25min, 17-20% A, 25-40min, 20% A, 40-45min, 20-28% A, 45-48min, 28-35% A, 48-60min, 35% A.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810103484.0A CN108362809B (en) | 2018-02-01 | 2018-02-01 | Quality evaluation method of ginkgo leaf and extract and preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810103484.0A CN108362809B (en) | 2018-02-01 | 2018-02-01 | Quality evaluation method of ginkgo leaf and extract and preparation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108362809A CN108362809A (en) | 2018-08-03 |
| CN108362809B true CN108362809B (en) | 2020-07-10 |
Family
ID=63004386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810103484.0A Expired - Fee Related CN108362809B (en) | 2018-02-01 | 2018-02-01 | Quality evaluation method of ginkgo leaf and extract and preparation thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108362809B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110824072B (en) * | 2019-12-18 | 2021-11-05 | 神威药业集团有限公司 | Method for constructing fingerprint of flavonoid in ginkgo leaf extract or preparation thereof |
| CN111537656B (en) * | 2020-06-19 | 2021-07-20 | 劲牌有限公司 | Identification method of tartary buckwheat extract |
| CN113995776B (en) * | 2020-07-27 | 2023-01-13 | 浙江康恩贝制药股份有限公司 | Ginkgo leaf flavone extract, preparation method thereof and characteristic map construction method |
| CN113533270A (en) * | 2021-06-16 | 2021-10-22 | 万邦德制药集团有限公司 | Quality control method of ginkgo leaf dripping pills |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110670A (en) * | 2012-10-23 | 2013-05-22 | 北京华润高科天然药物有限公司 | Preparation method for efficiently extracting separating high-purity flavone components from ginkgo leaf |
| CN104523771A (en) * | 2015-01-13 | 2015-04-22 | 中国药科大学 | Ginkgo leaf total flavones and preparing method thereof |
| CN106706809A (en) * | 2016-12-26 | 2017-05-24 | 河北神威药业有限公司 | Method for simultaneously determining contents of multiple components in Shuxuening injection |
| CN106770828A (en) * | 2016-12-26 | 2017-05-31 | 河北神威药业有限公司 | It is a kind of while the method for determining multicomponent content in ginkgo biloba p.e and its preparation |
| CN107064397A (en) * | 2017-06-16 | 2017-08-18 | 黑龙江珍宝岛药业股份有限公司 | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one |
-
2018
- 2018-02-01 CN CN201810103484.0A patent/CN108362809B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103110670A (en) * | 2012-10-23 | 2013-05-22 | 北京华润高科天然药物有限公司 | Preparation method for efficiently extracting separating high-purity flavone components from ginkgo leaf |
| CN104523771A (en) * | 2015-01-13 | 2015-04-22 | 中国药科大学 | Ginkgo leaf total flavones and preparing method thereof |
| CN106706809A (en) * | 2016-12-26 | 2017-05-24 | 河北神威药业有限公司 | Method for simultaneously determining contents of multiple components in Shuxuening injection |
| CN106770828A (en) * | 2016-12-26 | 2017-05-31 | 河北神威药业有限公司 | It is a kind of while the method for determining multicomponent content in ginkgo biloba p.e and its preparation |
| CN107064397A (en) * | 2017-06-16 | 2017-08-18 | 黑龙江珍宝岛药业股份有限公司 | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one |
Non-Patent Citations (4)
| Title |
|---|
| Rapid and Sensitive Determination of Major Active Ingredients and Toxic Components in Leaves Extract (EGb 761) by a Validated UPLC–MS-MS Method;Yao Wang et al;《Journal of Chromatographic Science》;20170109;第55卷(第4期);第459-464页 * |
| 一测多评法同时测定舒血宁注射液中6种黄酮类成分;张方 等;《中国药师》;20151231;第18卷(第10期);第1652-1656页 * |
| 一测多评法测定银杏叶提取物中7种黄酮类成分含量;杨艳模 等;《中国实验方剂学杂志》;20140430;第20卷(第8期);第82-85页 * |
| 指纹图谱结合一测多评模式在中药鱼腥草质量评价中的应用研究;何兵 等;《中国中药杂志》;20130831;第38卷(第16期);第2682-2689页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108362809A (en) | 2018-08-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108362809B (en) | Quality evaluation method of ginkgo leaf and extract and preparation thereof | |
| CN106770828B (en) | Method that is a kind of while measuring multicomponent content in ginkgo biloba p.e and its preparation | |
| CN105467059B (en) | A kind of quality determining method for the Chinese medicine composition for treating blood urine | |
| CN106706809B (en) | Method that is a kind of while measuring multicomponent content in Shu Xuening injection | |
| CN108896673B (en) | Method for determining content of chlorogenic acid, luteolin and apigenin in spider fruits | |
| CN107884483B (en) | Method for measuring content of flavonoid component in ginkgo leaf and preparation thereof and application | |
| CN107064397A (en) | A kind of Shu Xuening injection method of quality control that many evaluation amount methods and finger-print are surveyed based on one | |
| CN107037167A (en) | The method of flavones ingredient content in multi-target ingredient quantitative determination ginkgo leaf | |
| CN106442789A (en) | Establishment and active component quantitative analysis methods of compound Xuezhining extract fingerprint map | |
| CN104764820A (en) | Method for determining content of active ingredients such as ephedrine hydrochloride and pseudoephedrine hydrochloride in pinellia ternata syrup | |
| CN110927311B (en) | Construction method of UPLC (ultra performance liquid chromatography) characteristic spectrum of dogbane leaf medicinal material and method for measuring content of flavonoid component of dogbane leaf medicinal material | |
| CN109991327A (en) | One surveys the methods for commenting method evaluation field thistle quality more | |
| CN109521103A (en) | A kind of three Le slurry oral solution quality determining method having both qualitative and quantitative evaluation | |
| CN112730674B (en) | Quality detection method of momordica grosvenori tea | |
| CN110568111B (en) | Method for detecting oligosaccharide in morinda officinalis formula particles | |
| CN105004833A (en) | Detection method for traditional Chinese medicine preparation for treating acute gouty arthritis and gout | |
| CN104634911B (en) | A kind of 4 kinds of flavonoids effective constituent detection methods of CHUANKEZHI ZHUSHEYE | |
| CN106124682A (en) | A kind of composition method of inspection of Radix Et Caulis Acanthopanacis Senticosi injection | |
| CN107941955B (en) | The method of 9 kinds of functional active components in a kind of while quick detection YEDAO LUGUI JIU | |
| CN109900829A (en) | A kind of method of quality control of particle for relieving heat and cough | |
| CN113567593B (en) | Ginkgo honey ring oral solution multi-component content detection and fingerprint spectrum method construction thereof | |
| CN110687224B (en) | Method for measuring triptolide A in tripterygium wilfordii medicinal material and tripterygium wilfordii multi-glycoside tablet prepared from tripterygium wilfordii medicinal material | |
| CN104597168B (en) | Ramulus Mori refines decoction pieces content assaying method | |
| CN103512979A (en) | Detection method of pharmaceutical composition Zuozhudaxi | |
| CN108717088B (en) | Method for evaluating volatile components of radix curcumae medicinal material by one test and multiple tests |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200710 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |