CN112147265A - 一种金银花抗炎质量标志物筛选、品质鉴定方法及应用 - Google Patents
一种金银花抗炎质量标志物筛选、品质鉴定方法及应用 Download PDFInfo
- Publication number
- CN112147265A CN112147265A CN202010981475.9A CN202010981475A CN112147265A CN 112147265 A CN112147265 A CN 112147265A CN 202010981475 A CN202010981475 A CN 202010981475A CN 112147265 A CN112147265 A CN 112147265A
- Authority
- CN
- China
- Prior art keywords
- honeysuckle
- inflammatory
- screening
- quality
- markers
- 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.)
- Pending
Links
- 230000003110 anti-inflammatory effect Effects 0.000 title claims abstract description 62
- 238000000034 method Methods 0.000 title claims abstract description 37
- 238000012216 screening Methods 0.000 title claims abstract description 32
- 239000003550 marker Substances 0.000 title claims description 13
- 241000205585 Aquilegia canadensis Species 0.000 title abstract 5
- HEYZWPRKKUGDCR-WRMJXEAJSA-N Swertiamarin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@H]1[C@@H](C=C)[C@@]2(O)C(C(=O)OCC2)=CO1 HEYZWPRKKUGDCR-WRMJXEAJSA-N 0.000 claims abstract description 32
- HEYZWPRKKUGDCR-QBXMEVCASA-N Swertiamarin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](C=C)[C@]2(O)CCOC(=O)C2=CO1 HEYZWPRKKUGDCR-QBXMEVCASA-N 0.000 claims abstract description 32
- 239000000178 monomer Substances 0.000 claims abstract description 25
- CWVRJTMFETXNAD-FWCWNIRPSA-N 3-O-Caffeoylquinic acid Natural products O[C@H]1[C@@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-FWCWNIRPSA-N 0.000 claims abstract description 24
- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 claims abstract description 24
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 claims abstract description 24
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 claims abstract description 24
- 229940074393 chlorogenic acid Drugs 0.000 claims abstract description 24
- FFQSDFBBSXGVKF-KHSQJDLVSA-N chlorogenic acid Natural products O[C@@H]1C[C@](O)(C[C@@H](CC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O FFQSDFBBSXGVKF-KHSQJDLVSA-N 0.000 claims abstract description 24
- 235000001368 chlorogenic acid Nutrition 0.000 claims abstract description 24
- BMRSEYFENKXDIS-KLZCAUPSSA-N cis-3-O-p-coumaroylquinic acid Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)cc2)[C@@H]1O)C(=O)O BMRSEYFENKXDIS-KLZCAUPSSA-N 0.000 claims abstract description 24
- 238000002474 experimental method Methods 0.000 claims abstract description 21
- 239000000463 material Substances 0.000 claims abstract description 18
- 230000000694 effects Effects 0.000 claims abstract description 17
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims abstract description 12
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 claims abstract description 12
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 235000009498 luteolin Nutrition 0.000 claims abstract description 12
- 241001570521 Lonicera periclymenum Species 0.000 claims description 102
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 claims description 31
- 239000003814 drug Substances 0.000 claims description 26
- AMBQHHVBBHTQBF-UHFFFAOYSA-N Loganin Natural products C12C(C)C(O)CC2C(C(=O)OC)=COC1OC1OC(CO)C(O)C(O)C1O AMBQHHVBBHTQBF-UHFFFAOYSA-N 0.000 claims description 25
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- 230000005200 bud stage Effects 0.000 claims description 13
- AMBQHHVBBHTQBF-UOUCRYGSSA-N loganin Chemical compound O([C@@H]1OC=C([C@H]2C[C@H](O)[C@H](C)[C@H]21)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O AMBQHHVBBHTQBF-UOUCRYGSSA-N 0.000 claims description 13
- JNNGEAWILNVFFD-UHFFFAOYSA-N epiloganic acid Natural products CC1C(O)CC(C(=CO2)C(O)=O)C1C2OC1OC(CO)C(O)C(O)C1O JNNGEAWILNVFFD-UHFFFAOYSA-N 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 10
- JNNGEAWILNVFFD-DXNVVGTLSA-N 7-Epiloganic acid Chemical compound O([C@H]1[C@H]2[C@@H](C(=CO1)C(O)=O)C[C@@H](O)[C@@H]2C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O JNNGEAWILNVFFD-DXNVVGTLSA-N 0.000 claims description 9
- WMQIBEYREGDWOS-UOUCRYGSSA-N Loganic acid Natural products CC[C@H]1O[C@@H](O[C@@H]2OC=C([C@H]3C[C@H](O)[C@H](C)[C@@H]23)C(=O)O)[C@H](O)[C@@H](O)[C@@H]1O WMQIBEYREGDWOS-UOUCRYGSSA-N 0.000 claims description 9
- 239000003643 water by type Substances 0.000 claims description 9
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 8
- VSJGJMKGNMDJCI-ZASXJUAOSA-N Sweroside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](C=C)[C@H](CCOC2=O)C2=CO1 VSJGJMKGNMDJCI-ZASXJUAOSA-N 0.000 claims description 7
- 238000010828 elution Methods 0.000 claims description 7
- VSJGJMKGNMDJCI-QXSNVGMTSA-N sweroside Natural products OC[C@H]1O[C@H](O[C@@H]2OC=C3[C@@H](CCOC3=O)[C@H]2C=C)[C@H](O)[C@@H](O)[C@@H]1O VSJGJMKGNMDJCI-QXSNVGMTSA-N 0.000 claims description 7
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims description 7
- 238000004807 desolvation Methods 0.000 claims description 6
- 238000000132 electrospray ionisation Methods 0.000 claims description 6
- 238000011156 evaluation Methods 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 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 4
- 235000019253 formic acid Nutrition 0.000 claims description 4
- 238000000513 principal component analysis Methods 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- SAJHQVLFIJLYPM-UHFFFAOYSA-N 8-Epiloganin Natural products COC(O)C1=COC(OC2OC(CO)C(O)C(O)C2O)C3C(C)C(O)CC13 SAJHQVLFIJLYPM-UHFFFAOYSA-N 0.000 claims description 3
- 241000245240 Lonicera Species 0.000 claims description 3
- 238000001514 detection method Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 239000004047 hole gas Substances 0.000 claims description 3
- AMBQHHVBBHTQBF-OZIIXKNCSA-N methyl (1s,4as,6s,7s,7as)-6-hydroxy-7-methyl-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,4a,5,6,7,7a-hexahydrocyclopenta[c]pyran-4-carboxylate Chemical compound O([C@@H]1OC=C([C@H]2C[C@H](O)[C@@H](C)[C@H]21)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O AMBQHHVBBHTQBF-OZIIXKNCSA-N 0.000 claims description 3
- -1 secoxylogenin Chemical compound 0.000 claims description 3
- CSKKDSFETGLMSB-NRZPKYKESA-N (-)-secologanin Chemical compound C=C[C@@H]1[C@H](CC=O)C(C(=O)OC)=CO[C@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CSKKDSFETGLMSB-NRZPKYKESA-N 0.000 claims description 2
- JNNGEAWILNVFFD-OZFVSFPVSA-N (1s,4as,6s,7s,7as)-6-hydroxy-7-methyl-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,4a,5,6,7,7a-hexahydrocyclopenta[c]pyran-4-carboxylic acid Chemical compound O([C@H]1[C@H]2[C@@H](C(=CO1)C(O)=O)C[C@H](O)[C@H]2C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O JNNGEAWILNVFFD-OZFVSFPVSA-N 0.000 claims description 2
- JNNGEAWILNVFFD-BFWCLYIISA-N 8-Epiloganic acid Natural products O=C(O)C=1[C@@H]2[C@H]([C@H](C)[C@@H](O)C2)[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O2)OC=1 JNNGEAWILNVFFD-BFWCLYIISA-N 0.000 claims description 2
- CSKKDSFETGLMSB-FUJZYWHJSA-N Secologanin Natural products C=C[C@@H]1[C@H](CC=O)C(C(=O)OC)=CO[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CSKKDSFETGLMSB-FUJZYWHJSA-N 0.000 claims description 2
- 238000004949 mass spectrometry Methods 0.000 claims description 2
- 238000007781 pre-processing Methods 0.000 claims description 2
- 238000012360 testing method Methods 0.000 claims 1
- 238000001228 spectrum Methods 0.000 abstract description 10
- 238000002366 time-of-flight method Methods 0.000 abstract description 10
- 102000003945 NF-kappa B Human genes 0.000 abstract description 8
- 108010057466 NF-kappa B Proteins 0.000 abstract description 8
- 239000005089 Luciferase Substances 0.000 abstract description 7
- 238000013441 quality evaluation Methods 0.000 abstract description 3
- 239000012071 phase Substances 0.000 description 10
- 239000000126 substance Substances 0.000 description 8
- 238000010586 diagram Methods 0.000 description 7
- 238000011160 research Methods 0.000 description 6
- 238000000605 extraction Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 241000628997 Flos Species 0.000 description 4
- 241000100289 Lonicera confusa Species 0.000 description 4
- 244000167230 Lonicera japonica Species 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- 238000002137 ultrasound extraction Methods 0.000 description 4
- 235000017617 Lonicera japonica Nutrition 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003181 biological factor Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000003205 fragrance Substances 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000208828 Caprifoliaceae Species 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 241000282994 Cervidae Species 0.000 description 1
- 201000000297 Erysipelas Diseases 0.000 description 1
- 208000015220 Febrile disease Diseases 0.000 description 1
- 206010017553 Furuncle Diseases 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 108090001007 Interleukin-8 Proteins 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000003255 drug test Methods 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- MWLKXILGJPSPKZ-LWSBBEHMSA-N methyl (2S,3R,4S)-3-ethenyl-4-[(Z)-3-[(2S,3R,4R)-3-ethenyl-5-methoxycarbonyl-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,4-dihydro-2H-pyran-4-yl]-4-oxobut-2-enyl]-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,4-dihydro-2H-pyran-5-carboxylate Chemical compound O([C@@H]1OC=C([C@H]([C@H]1C=C)C\C=C(/C=O)[C@@H]1C(=CO[C@@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@@H]1C=C)C(=O)OC)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O MWLKXILGJPSPKZ-LWSBBEHMSA-N 0.000 description 1
- 238000000874 microwave-assisted extraction Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 229940126672 traditional medicines Drugs 0.000 description 1
- 238000001195 ultra high performance liquid chromatography Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000012795 verification Methods 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
- G01N33/5023—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects on expression patterns
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Library & Information Science (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明利用293t细胞及NF‑kB双荧光素酶报告系统做抗炎活性实验,结果表明金银花的抗炎活性差异较大,选择活性最优指标做活性抗炎单体的筛选。通过UPLC‑Q/TOF结合NF‑kB双荧光素酶报告系统,将金银花中各个峰接出做活性筛选实验。结果筛出4个活性单体,并对这4个峰进行解谱鉴定,确定分别为绿原酸、木犀草苷、獐芽菜苷和獐牙菜苦苷,将这4个抗炎指标成分作为金银花多指标质量评价的物质基础。本文首先利用药典方法对来自不同产地的金银花样品进行鉴定,并结合UPLC‑Q/TOF方法进行进一步分析。
Description
技术领域
本发明涉及天然中药材的鉴定方法,具体的涉及一种金银花抗炎质量标志物筛选、品质鉴定方法及应用。
背景技术
金银花为忍冬科忍冬属多年生半常绿木本植物忍冬(Lonicera japonicaThunb.)的干燥花蕾或带初开的花,它性甘寒气芳香,甘寒清热而不伤胃,芳香透达又可祛邪,自古以来被誉为清热解毒的良药,并有“中药抗生素”之称。金银花是国务院确定的70种名贵药材之一,也是第一批纳入“药食同源”名录的中药。金银花原产于我国大陆,大部分北方省份均有分布,主产地有河北巨鹿、山东平邑、河南封丘等。市场上种植的品种较多,有巨花一号、大毛花、树形四季花、北花一号、九丰一号、豫封一号、金花三号等。“金银花”一名出自《本草纲目》,由于忍冬花初开为白色,后转为黄色,因此得名金银花。在中国的历代医学文献中,均发现有金银花及其药效的记载。《中国药典》2015年版收载的金银花功效为:清热解毒,疏散风热。用于痈肿疔疮,喉痹,丹毒,热血毒痢,风热感冒,温病发热。
中药材指纹图谱是一种对中药质量进行评价的切实可行的方法,是将某种或是某产地的中药经过预处理后,运用现代分析技术对中药化学信息以图像(图形)的形式进行表征并加以描述,得到能够标示该中药特性的某类或数类成分图谱。中药指纹图谱的主要特点:通过特征指纹图谱能有效识别样品的产地或真伪;通过指纹峰峰面积或比例的测定,能有效控制样品的质量,确保样品批次之间的相对稳定。所以,在功效成分不完全明确时制定中药的指纹图谱,对于有效控制中药的质量非常有必要。
目前,指纹图谱己成为国际公认的控制中药材或中成药质量的有效手段。涉及中药指纹图谱技术方法有很多,包括TLC、HPLC、HPLC/MS、GC、GC/MS、HPCE、IR等。中药指纹图谱的基本特性是整体性和模糊性,中药指纹图谱的评价应考虑图谱的这两个方面,中药材含有多类化学成分,它们与中药的有效性和安全性有关,当具备了一定条件,中药成分就能成为反映中药质量的标志物(quality marker,Q-markers)。中药材的生物学因素或非生物学因素所形成的次生代谢物是其中药质量标志物的物质基础和发现药物作用的物质基础以及发现新药的重要来源。
发明内容
本发明利用293t细胞及NF-kB双荧光素酶报告系统做抗炎活性实验,结果表明金银花的抗炎活性差异较大,选择活性最优指标做活性抗炎单体的筛选。通过UPLC-Q/TOF结合NF-kB双荧光素酶报告系统,将金银花中各个峰接出做活性筛选实验。结果筛出4个活性单体,并对这4个峰进行解谱鉴定,确定分别为绿原酸、木犀草苷、獐芽菜苷和獐牙菜苦苷,将这4个抗炎指标成分作为金银花多指标质量评价的物质基础。本文首先利用药典方法对来自不同产地的金银花样品进行鉴定,并结合UPLC-Q/TOF方法进行进一步分析。
为了实现上述发明目的,本发明采用的技术方案为:一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,包括一下步骤:
(1)将待检测的金银花按不同花期分类得到不同花期金银花;
(2)将步骤(1)中所述的不同花期金银花进行预处理,得到不同花期金银花待测样品;
(3)将步骤(2)得到的所述不同花期金银花待测样品采用超高效液相色谱仪串联四极杆/飞行时间质谱仪分离;
(4)将步骤(3)中得到的分离结果进行PCA分析,计算出贡献值大的若干makers;
(5)将步骤(2)得到的所述不同花期金银花待测样品进行抗炎活性实验,筛选出用于检测抗炎质量标志物组;
(6)将步骤(5)中得出所述用于检测抗炎质量标志物组,通过步骤(2)超高效液相色谱仪串联四极杆/飞行时间质谱仪分离,并按时间接收分离后样品;
(7)将步骤(6)中接收到的所述分离后样品进行抗炎活性实验,得到若干活性单体;
(8)将步骤(4)得到的若干makers和步骤(7)得到的若干活性单体进行抗炎活性实验,得到抗炎活性单体的相对含量;
(9)将步骤(8)中得到的獐芽菜苷和獐牙菜苦苷以及中药药典规定的绿原酸、木犀草苷作为金银花抗炎质量标志物。
所述步骤(2)中的色谱条件:色谱柱:Waters ACQUITYUPLCTM BEH C18(1.7μm,2.1×100mm);流动相:(A)乙腈,(B)0.1%(v/v)甲酸水溶液,梯度洗脱;流速为0.4mL·min-1;柱温:30℃;进样2μL。
所述步骤(2)中的质谱条件:电喷雾离子源(ESI),分别采用正离子(负离子)V模式下检测;脱溶剂气为600L/h,脱溶剂气温度为380℃,锥孔气为50L/h,离子源为100℃,毛细管电压正离子为3.0kV(负离子为2.5kV),锥孔电压为30V。0.1s(间隔0.02s)采集一次谱图;采用LEA([M+H]+=555.2931,[M+H]-=553.2775)作Lockmass。
所述洗脱梯度如下:
所述不同花期金银花分为绿蕾期、白蕾期、金花期和枝叶期。
所述步骤(2)所述预处理包括:将所述待检测的金银花干制,研磨至80目,25%甲醇水溶液250W,35kHz,40℃超声30min萃取即得。
所述步骤(4)得到的若干makers为:swertiamarin(獐芽菜苦苷),sweroside(獐芽菜苷),loganin(马钱子苷),8-epi-loganin(8位异构马钱子苷),8-epi-loganic acid(8位异构马钱苷酸),chlorogenic acid(绿原酸),loganic acid(马钱苷酸),secoxyloganin(断氧化马钱子苷),aldosecologanin。
所述步骤(5)中将所述不同花期金银花待测样品进行抗炎活性实验,其中绿蕾期活性最好,筛选出绿蕾期用于检测抗炎质量标志物组。
所述步骤(6)中所述时间为30秒,将绿蕾期金银花进行超高效液相色谱仪串联四极杆/飞行时间质谱仪分离,每30秒接一个样品,做活性筛选实验。
所述活性筛选实验得到绿蕾期中有6个活性单体,分别为swertiamarin(獐芽菜苦苷),sweroside(獐芽菜苷),secoxyloganin(断氧化马钱子苷),chlorogenic acid(绿原酸),loganic acid(马钱苷酸),aldosecologanin。
一种金银花药材品质鉴定方法,其特征在于,采用上述方法筛选出来的獐芽菜苷和獐牙菜苦苷、绿原酸、木犀草苷作为金银花抗炎质量标志物,建立金银花对照指纹图谱,通过中药色谱指纹图谱相似度评价系统比较待测金银花的指纹图谱与对照指纹图谱的相似度,从而判断金银花要求的品质。
一种金银花抗炎质量标志物筛选鉴定方法在金银花药材品质鉴定中的应用。
化学成分是中药质量控制的重要指标,其可测性也是中药质量标志物确定的重要依据。目前中药化学成分主要通过色谱技术手段来进行分析测定,金银花中化学成分结构复杂,利用色谱质谱联用技术对金银花质量标志进行鉴定及定量是金银花质量标志物研究的重要方向。本研究利用293t细胞及NF-kB双荧光素酶报告系统筛选抗炎活性成分,并通过UPLC-Q/TOF进行抗炎质量标志物的鉴定。结果筛出4个活性单体,并对这4个峰进行解谱鉴定,确定分别为绿原酸、木犀草苷、獐芽菜苷和獐牙菜苦苷,基本确认了金银花抗炎质量标志物筛选鉴定方法,对金银花抗炎活性的品质鉴定有着重要的意义。
通过比较甲醇、水、乙醇等不同极性的溶剂以及常规加热回流法、超声提取法、冷热浸提法、微波萃取法等提取方法对金银花中有效成分提取效果的影响,结果以甲醇为提取溶剂,采用超声法提取,提取效果好且方法简便易行、重现性好。此外,对超声提取时间及甲醇浓度进行优化,采用25%甲醇超声30min提取金银花样品中绿原酸、木犀草苷、獐芽菜苷和獐牙菜苦苷等多种有效成分,所得指纹图谱信息量较大,既有水溶性成分又有脂溶性成分,符合指纹图谱整体性要求,能够反映金银花样品的大部分信息。
金银花化学指纹图谱的确立,结果表明将筛出4个活性单体(绿原酸、木犀草苷、獐芽菜苷和獐牙菜苦苷)作为金银花抗炎质量标志物应用超高效液相色谱进行化学指纹图谱识别研究,结果显示指纹图谱相似度差异较大,说明该化学指纹图谱系统可用于判定金银花药材品质及其伪品的鉴定。
附图说明
图1 UPLC-Q/TOF上图为正模式总离子流图,下图为负总离子流图
图2 A为金银花不同花期的质谱BPI图,B为Score图,C为Loading图;1-6分别为loganic acid(马钱苷酸)、aldosecologanin、chlorogenic acid(绿原酸)、swertiamarin(獐芽菜苦苷)、secoxyloganin(断氧化马钱子苷)、6sweroside(獐芽菜苷)。
图3抗炎活性单体的相对含量图
图4 101批次金银花及其伪品UPLC指纹图谱相似度评价图
具体实施方式
为了阐述本发明的技术方案和技术目的,下面具体实施方式对本发明做进一步介绍。
1、实验准备
1.1样本采集通过对金银花和山银花产区及药材市场进行调研,获得具有一定代表性的样本101份。其中金银花样品收集自河北巨鹿县、河南封丘县、山东平邑县等地。
表1金银花样本信息列表
上述101份样本按花期、品类分类,按上述表中干制后研磨至80目,精密称取过80目筛的金银花粉末约0.1g于50mL锥形瓶中,精密加入25%甲醇水溶液10mL,称定重量,超声30min(250W,35kHz,40℃),放冷,补足减失的重量。摇匀,过0.45μm滤膜,即得。
1.2仪器超高效液相色谱仪串联四极杆/飞行时间质谱仪(Waters ACQUITYTMUPLC-Q/TOF Premier,MassLynx V4.1工作站);Waters ACQUITY UPLC H-Class Coresystem超高效液相色谱仪,Empower3色谱工作站,配置自动进样器、二极管阵列检测器、四元梯度泵。
1.3主要试剂标准品绿原酸(批号:110753-201817,96.80%),木樨草苷(批号:111720-201805,98.74%),均购自中国食品药品检定研究院。对照品獐牙菜苷(20mg,Z-009-131225,HPLC≥98%)、獐牙菜苦苷(20mg,Z-002-130427,HPLC≥98%)均购自成都瑞芬思有限公司。液相用水为Millipore超纯水;乙腈(Fisher,色谱纯);磷酸、乙醇为分析纯。
2、金银花UPLC-Q/TOF实验方法的建立
采用超高效液相色谱仪串联四极杆/飞行时间质谱仪(Waters ACQUITYTM UPLC-Q/TOF Premier,MassLynx V4.1工作站)建立了初步的金银花UPLC图谱,如图1。
色谱条件:色谱柱:Waters ACQUITY UPLCTM BEH C18(1.7μm,2.1×100mm);流动相:(A)乙腈,(B)0.1%(v/v)甲酸水溶液,梯度洗脱,程序见表2;流速为0.4mL·min-1;柱温:30℃;进样2μL。
质谱条件:电喷雾离子源(ESI),分别采用正离子(负离子)V模式下检测;脱溶剂气为600L/h,脱溶剂气温度为380℃,锥孔气为50L/h,离子源为100℃,毛细管电压正离子为3.0kV(负离子为2.5kV),锥孔电压为30V。0.1s(间隔0.02s)采集一次谱图;采用LEA([M+H]+=555.2931,[M+H]-=553.2775)作Lockmass。
表2流动相梯度洗脱表
3、实验室金银花抗炎活性研究
通过UPLC-Q/TOF做金银花不同花期的PCA(主成分分析):
图2A为金银花不同花期的质谱BPI图,B为Score图,C为Loading图,其中1-6分别为loganic acid(马钱苷酸)、aldosecologanin、chlorogenic acid(绿原酸)、swertiamarin(獐芽菜苦苷)、secoxyloganin(断氧化马钱子苷)、6sweroside(獐芽菜苷)。B图中四个花期的金银花分散度很高,说明四个花期金银花的特性相差很大,C图共计算出8个贡献值大的makers。分别为swertiamarin(獐芽菜苦苷),sweroside(獐芽菜苷),loganin(马钱子苷),8-epi-loganin(8位异构马钱子苷),8-epi-loganic acid(8位异构马钱苷酸),chlorogenic acid(绿原酸),loganic acid(马钱苷酸),secoxyloganin(断氧化马钱子苷),aldosecologanin。说明正是这些markers在不同花期的含量不同导致各花期差异较大。
4、不同花期金银花的抗炎活性比较
利用293t细胞及NF-kB双荧光素酶报告系统做抗炎活性实验,结果表明不同花期金银花的抗炎活性差异较大,只有绿蕾期活性最好。选择活性最好的绿蕾期做活性抗炎单体的筛选。利用UPLC-Q/TOF结合NF-kB双荧光素酶报告系统,将绿蕾中各个峰接出(每30秒接一个),做活性筛选实验。结果筛出绿蕾中有6个活性单体。对这6个峰进行解谱鉴定,确定分别为swertiamarin(獐芽菜苦苷),sweroside(獐芽菜苷),secoxyloganin(断氧化马钱子苷),chlorogenic acid(绿原酸),loganic acid(马钱苷酸),aldosecologanin。
选swertiamarin,sweroside代表性的单体买标准品,做抗炎活性实验的验证。包括NF-kB表达以及IL-6,IL-8等炎症因子的表达。做6个markers和6个活性单体(共10个)的相对含量图(热图),如图3所示。
将10个化合物在不同花期中的峰面积从MassLynx软件导出,将所有峰面积归一化,导入Cluster软件得到10个峰在所有样品中的相对含量。结果表明,有抗炎活性的单体在绿蕾中的相对含量最高,在其它花期很少,与前期各个花期金银花中绿蕾抗炎活性最好的结果一致。综合实验结果说明:不同花期金银花的抗炎活性有很大差异,是因为几个抗炎活性单体在不同花期中的含量不同导致。这对金银花抗炎活性的品质鉴定有着重要的意义,选取4个抗炎指标成分作为质量标志物进行金银花多指标质量评价的物质基础。
5、金银花指纹图谱的研究
对照品溶液的制备:精密称取对照品绿原酸、木犀草苷、獐牙菜苷和獐牙菜苦苷适量,用25%甲醇水溶液配制成单个对照品储备液,稀释得到标准使用液浓度依次为0.220mg/mL、0.008mg/mL、0.274mg/mL、0.258mg/mL。
供试品溶液的制备:精密称取过80目筛的金银花粉末约0.1g于50mL锥形瓶中,精密加入25%甲醇水溶液10mL,称定重量,超声30min(250W,35kHz,40℃),放冷,补足减失的重量。摇匀,过0.45μm滤膜,即得。
色谱条件:Waters H-Class超高效液相色谱仪,Waters ACQUITY UPLCTM BEH C18色谱柱(1.7μm,2.1×100mm);流动相:(A)乙腈,(B)0.1%(v/v)甲酸水溶液,梯度洗脱,程序见表2;流速为0.4mL·min-1;柱温:25℃;检测波长240nm;进样量2μL。
6、金银花指纹图谱的建立及共有指纹峰的确定
精密吸取绿原酸、木犀草苷、獐牙菜苷和獐牙菜苦苷混合对照品溶液和制备的101批供试品溶液各2μL,按照上述分析的色谱方法,分别注入Waters ACQUITYUPLC H-ClassCore system超高效液相色谱仪,记录35min的色谱图。将101批金银花样品色谱指纹图谱AIA数据文件(*.Cdf格式)导入国家药典委员会开发的“中药色谱指纹图谱相似度评价系统”,计算各指纹图谱间相似度。选取色谱图中13个较强峰,由数据分析系统辅助自动峰匹配,确定8个共有峰。以獐牙菜苷峰为参照峰(S峰),通过S峰的保留时间和成分峰面积积分值作参照,计算各共有峰的相对保留时间和相对峰积分面积,考察101批样品中相同成分峰相对峰面积的RSD值,符合指纹图谱的技术要求。
根据“中药注射剂色谱指纹图谱实验研究技术指南(试行)”,采用“中药色谱指纹图谱相似度评价系统2012年版(国家药典委员会)”对69批金银花的指纹图谱进行相似度分析处理,以中位数法生成69批金银花的对照指纹图谱,结果显示69批金银花(忍冬)与对照指纹图谱相似度为0.962~0.998之间,药典收载忍冬作为金银花合法品种;32批次的山银花与对照指纹图谱相似度为0.745~0.851之间,因与忍冬属不同种,功效成分存在一定差异,药理疗效也不同,药典收载为山银花以与金银花相区别。101批金银花及其伪品山银花UPLC指纹图谱叠加图如图4所示。
以上显示和描述了本发明的基本原理和主要特征和本发明的优点。本行业的技术人员应该了解,本发明不受上述实施例的限制,上述实施例和说明书中描述的只是说明本发明的原理,在不脱离本发明精神和范围的前提下,本发明还会有各种变化和改进,这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。
Claims (12)
1.一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,包括一下步骤:
(1)将待检测的金银花按不同花期分类得到不同花期金银花;
(2)将步骤(1)中所述的不同花期金银花进行预处理,得到不同花期金银花待测样品;
(3)将步骤(2)得到的所述不同花期金银花待测样品采用超高效液相色谱仪串联四极杆/飞行时间质谱仪分离;
(4)将步骤(3)中得到的分离结果进行PCA分析,计算出贡献值大的若干makers;
(5)将步骤(2)得到的所述不同花期金银花待测样品进行抗炎活性实验,筛选出用于检测抗炎质量标志物组;
(6)将步骤(5)中得出所述用于检测抗炎质量标志物组,通过步骤(2)超高效液相色谱仪串联四极杆/飞行时间质谱仪分离,并按时间接收分离后样品;
(7)将步骤(6)中接收到的所述分离后样品进行抗炎活性实验,得到若干活性单体;
(8)将步骤(4)得到的若干makers和步骤(7)得到的若干活性单体进行抗炎活性实验,得到抗炎活性单体的相对含量;
(9)将步骤(8)中得到的獐芽菜苷和獐牙菜苦苷以及中药药典规定的绿原酸、木犀草苷作为金银花抗炎质量标志物。
2.根据权利要求1所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述步骤(2)中的色谱条件:色谱柱:Waters ACQUITYUPLCTM BEH C18(1.7μm,2.1×100mm);流动相:(A)乙腈,(B)0.1%(v/v)甲酸水溶液,梯度洗脱;流速为0.4mL·min-1;柱温:30℃;进样2μL。
3.根据权利要求1所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述步骤(2)中的质谱条件:电喷雾离子源(ESI),分别采用正离子(负离子)V模式下检测;脱溶剂气为600L/h,脱溶剂气温度为380℃,锥孔气为50L/h,离子源为100℃,毛细管电压正离子为3.0kV(负离子为2.5kV),锥孔电压为30V。0.1s(间隔0.02s)采集一次谱图;采用LEA([M+H]+=555.2931,[M+H]-=553.2775)作Lockmass。
5.根据权利要求1所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述不同花期金银花分为绿蕾期、白蕾期、金花期和枝叶期。
6.根据权利要求1所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述步骤(2)所述预处理包括:将所述待检测的金银花干制,研磨至80目,25%甲醇水溶液250W,35kHz,40℃超声30min萃取即得。
7.根据权利要求1所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述步骤(4)得到的若干makers为:swertiamarin(獐芽菜苦苷),sweroside(獐芽菜苷),loganin(马钱子苷),8-epi-loganin(8位异构马钱子苷),8-epi-loganic acid(8位异构马钱苷酸),chlorogenic acid(绿原酸),loganic acid(马钱苷酸),secoxyloganin(断氧化马钱子苷),aldosecologanin。
8.根据权利要求1所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述步骤(5)中将所述不同花期金银花待测样品进行抗炎活性实验,其中绿蕾期活性最好,筛选出绿蕾期用于检测抗炎质量标志物组。
9.根据权利要求8所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述步骤(6)中所述时间为30秒,将绿蕾期金银花进行超高效液相色谱仪串联四极杆/飞行时间质谱仪分离,每30秒接一个样品,做活性筛选实验。
10.根据权利要求9所述的一种金银花抗炎质量标志物筛选鉴定方法,其特征在于,所述活性筛选实验得到绿蕾期中有6个活性单体,分别为swertiamarin(獐芽菜苦苷),sweroside(獐芽菜苷),secoxyloganin(断氧化马钱子苷),chlorogenic acid(绿原酸),loganic acid(马钱苷酸),aldosecologanin。
11.一种金银花药材品质鉴定方法,其特征在于,采用如权利要求1-10任意一项所述方法筛选出来的獐芽菜苷和獐牙菜苦苷、绿原酸、木犀草苷作为金银花抗炎质量标志物,建立金银花对照指纹图谱,通过中药色谱指纹图谱相似度评价系统比较待测金银花的指纹图谱与对照指纹图谱的相似度,从而判断金银花要求的品质。
12.一种金银花抗炎质量标志物筛选鉴定方法在金银花药材品质鉴定中的应用,其特征在于,采用如权利要求1-10任意一项所述方法,鉴定金银花药材品质。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010981475.9A CN112147265A (zh) | 2020-09-17 | 2020-09-17 | 一种金银花抗炎质量标志物筛选、品质鉴定方法及应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010981475.9A CN112147265A (zh) | 2020-09-17 | 2020-09-17 | 一种金银花抗炎质量标志物筛选、品质鉴定方法及应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN112147265A true CN112147265A (zh) | 2020-12-29 |
Family
ID=73893914
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010981475.9A Pending CN112147265A (zh) | 2020-09-17 | 2020-09-17 | 一种金银花抗炎质量标志物筛选、品质鉴定方法及应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112147265A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113917058A (zh) * | 2021-09-30 | 2022-01-11 | 上海中科新生命生物科技有限公司 | 一种检测金银花药材中12种有效成分的方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104020235A (zh) * | 2014-06-26 | 2014-09-03 | 广西壮族自治区药用植物园 | 一种同时测定金银花中绿原酸和木犀草苷含量的方法 |
CN105842373A (zh) * | 2016-06-15 | 2016-08-10 | 华润三九医药股份有限公司 | 一种建立金银花的药物制剂的指纹图谱的方法 |
CN111443153A (zh) * | 2020-05-11 | 2020-07-24 | 贵州中医药大学 | 一种基于谱效关系的金银花质量检测方法 |
-
2020
- 2020-09-17 CN CN202010981475.9A patent/CN112147265A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104020235A (zh) * | 2014-06-26 | 2014-09-03 | 广西壮族自治区药用植物园 | 一种同时测定金银花中绿原酸和木犀草苷含量的方法 |
CN105842373A (zh) * | 2016-06-15 | 2016-08-10 | 华润三九医药股份有限公司 | 一种建立金银花的药物制剂的指纹图谱的方法 |
CN111443153A (zh) * | 2020-05-11 | 2020-07-24 | 贵州中医药大学 | 一种基于谱效关系的金银花质量检测方法 |
Non-Patent Citations (3)
Title |
---|
MIN JIANG ET AL.: "The Screening Research of Anti-Inflammatory Bioactive Markers from Different Flowering Phases of Flos Lonicerae Japonicae", 《PLOS ONE》 * |
代云桃 等: "中药饮片标准汤剂的质量评价案例――金银花", 《中国中药杂志》 * |
刘征辉 等: "金银花中抗炎活性成分獐牙菜苷和獐牙菜苦苷定量方法的研究", 《中国现代中药》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113917058A (zh) * | 2021-09-30 | 2022-01-11 | 上海中科新生命生物科技有限公司 | 一种检测金银花药材中12种有效成分的方法 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109444290B (zh) | 车前草药材uplc特征图谱的构建方法和检测方法 | |
Kim et al. | Chemical fingerprinting of Codonopsis pilosula and simultaneous analysis of its major components by HPLC–UV | |
CN109374785B (zh) | 淡竹叶药材uplc特征图谱的构建方法和检测方法 | |
CN113866293A (zh) | 一种快速识别分析洋甘菊中化学成分的方法 | |
CN110031564B (zh) | 基于hplc指纹图谱的天然植物抗球虫饲料添加剂的质量检测方法 | |
CN109632995B (zh) | 一种酸枣仁黄酮类成分uplc指纹图谱的建立方法及应用 | |
CN112557563A (zh) | 一种江西道地药材枳壳的鉴别方法 | |
CN112147265A (zh) | 一种金银花抗炎质量标志物筛选、品质鉴定方法及应用 | |
CN113702557A (zh) | 一种具有止血凝血作用的裸花紫珠有效成分的评价方法 | |
CN113945674B (zh) | 一种地黄炮制品的特征图谱及分析方法 | |
CN112946132B (zh) | 月季花药材指纹图谱及其构建方法和月季花药材的质量检测方法 | |
CN114814057A (zh) | 一种非靶向代谢组学区分卷柏品种真伪的方法及应用 | |
CN109374787B (zh) | 对叶百部药材uplc特征图谱的构建方法和检测方法 | |
CN110441443B (zh) | 一种有柄石韦、石韦、庐山石韦与华北石韦的uplc特征图谱构建方法及其鉴别方法 | |
CN113588855A (zh) | 野马追的质量检测方法 | |
CN110297047B (zh) | 细梗香草药材的hplc指纹图谱的建立方法及其指纹图谱 | |
CN113640432A (zh) | 一种壮腰健身丸的质量评价方法 | |
CN107894466B (zh) | 一种金桑芪抗毒制剂hplc指纹图谱的测定方法及金桑芪抗毒制剂质量控制方法 | |
CN104133028B (zh) | 一种茜草配方颗粒高效液相色谱指纹图谱的建立方法 | |
CN114216978B (zh) | 野马追配方颗粒指纹图谱的构建方法及其标准指纹图谱和应用 | |
CN112240914A (zh) | 不同外观表型金线莲中黄酮成分的检测方法 | |
CN110376311B (zh) | 用于鉴别当归与混伪品东当归的方法 | |
CN114965842B (zh) | 口炎清整体质量检测方法及应用 | |
CN113759026B (zh) | 伸筋草及其制剂特征图谱及其构建方法 | |
CN114720602B (zh) | 石见穿药材或其标准汤剂的特征图谱构建方法和检测方法 |
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 | ||
WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20201229 |
|
WD01 | Invention patent application deemed withdrawn after publication |