CN107315061B - A kind of detection method of alizarin root of Dahurian angelica Chinese materia medica preparation that treating uterus bleeding - Google Patents
A kind of detection method of alizarin root of Dahurian angelica Chinese materia medica preparation that treating uterus bleeding Download PDFInfo
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- CN107315061B CN107315061B CN201710452403.3A CN201710452403A CN107315061B CN 107315061 B CN107315061 B CN 107315061B CN 201710452403 A CN201710452403 A CN 201710452403A CN 107315061 B CN107315061 B CN 107315061B
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- alizarin
- dahurian angelica
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- RGCKGOZRHPZPFP-UHFFFAOYSA-N alizarin Chemical compound C1=CC=C2C(=O)C3=C(O)C(O)=CC=C3C(=O)C2=C1 RGCKGOZRHPZPFP-UHFFFAOYSA-N 0.000 title claims abstract description 132
- 238000002360 preparation method Methods 0.000 title claims abstract description 108
- 241000213006 Angelica dahurica Species 0.000 title claims abstract description 80
- 239000002398 materia medica Substances 0.000 title claims abstract description 65
- 230000000740 bleeding effect Effects 0.000 title claims abstract description 12
- 210000004291 uterus Anatomy 0.000 title claims abstract description 12
- 238000001514 detection method Methods 0.000 title claims description 19
- 239000003814 drug Substances 0.000 claims abstract description 53
- 238000000034 method Methods 0.000 claims abstract description 51
- 239000000463 material Substances 0.000 claims abstract description 44
- 241000123069 Ocyurus chrysurus Species 0.000 claims abstract description 40
- 235000003143 Panax notoginseng Nutrition 0.000 claims abstract description 36
- 241000180649 Panax notoginseng Species 0.000 claims abstract description 36
- OLOOJGVNMBJLLR-UHFFFAOYSA-N imperatorin Chemical compound C1=CC(=O)OC2=C1C=C1C=COC1=C2OCC=C(C)C OLOOJGVNMBJLLR-UHFFFAOYSA-N 0.000 claims abstract description 33
- XKVWLLRDBHAWBL-UHFFFAOYSA-N imperatorin Natural products CC(=CCOc1c2OCCc2cc3C=CC(=O)Oc13)C XKVWLLRDBHAWBL-UHFFFAOYSA-N 0.000 claims abstract description 33
- 241000125175 Angelica Species 0.000 claims abstract description 28
- 235000001287 Guettarda speciosa Nutrition 0.000 claims abstract description 28
- BBNQQADTFFCFGB-UHFFFAOYSA-N purpurin Chemical compound C1=CC=C2C(=O)C3=C(O)C(O)=CC(O)=C3C(=O)C2=C1 BBNQQADTFFCFGB-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229940079593 drug Drugs 0.000 claims abstract description 13
- VLGATXOTCNBWIT-UHFFFAOYSA-N rubimaillin Chemical compound O1C(C)(C)C=CC2=C1C1=CC=CC=C1C(O)=C2C(=O)OC VLGATXOTCNBWIT-UHFFFAOYSA-N 0.000 claims abstract description 13
- 238000012850 discrimination method Methods 0.000 claims abstract description 9
- NEFYSBQJYCICOG-YSEUJXISSA-N cyasterone Chemical compound C[C@H]1OC(=O)[C@@H](C)[C@@H]1C[C@@H](O)[C@](C)(O)[C@@H]1[C@@]2(C)CC[C@@H]3[C@@]4(C)C[C@H](O)[C@H](O)C[C@H]4C(=O)C=C3[C@]2(O)CC1 NEFYSBQJYCICOG-YSEUJXISSA-N 0.000 claims abstract description 8
- SXIFCLOUSXMYIX-UHFFFAOYSA-N cyasterone Natural products CC1OC(=O)C(C)C1CCC(C)(O)C2CCC3(O)C4=CC(=O)C5CC(O)C(O)CC5(C)C4CCC23C SXIFCLOUSXMYIX-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229930189092 Notoginsenoside Natural products 0.000 claims abstract description 3
- 229930182494 ginsenoside Natural products 0.000 claims abstract description 3
- 229940089161 ginsenoside Drugs 0.000 claims abstract description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 189
- 239000000243 solution Substances 0.000 claims description 175
- 239000013558 reference substance Substances 0.000 claims description 137
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 87
- 239000012085 test solution Substances 0.000 claims description 85
- 238000004587 chromatography analysis Methods 0.000 claims description 81
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 73
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 70
- 239000000523 sample Substances 0.000 claims description 61
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 60
- 239000002904 solvent Substances 0.000 claims description 53
- 239000000047 product Substances 0.000 claims description 37
- 239000000741 silica gel Substances 0.000 claims description 37
- 229910002027 silica gel Inorganic materials 0.000 claims description 37
- 238000012360 testing method Methods 0.000 claims description 37
- 239000003208 petroleum Substances 0.000 claims description 35
- 239000000706 filtrate Substances 0.000 claims description 33
- 235000019441 ethanol Nutrition 0.000 claims description 32
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 28
- 238000001914 filtration Methods 0.000 claims description 28
- BDAGIHXWWSANSR-UHFFFAOYSA-N formic acid Substances OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 26
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 24
- 239000007921 spray Substances 0.000 claims description 21
- 235000008216 herbs Nutrition 0.000 claims description 19
- 239000012488 sample solution Substances 0.000 claims description 19
- 239000000843 powder Substances 0.000 claims description 18
- 239000000377 silicon dioxide Substances 0.000 claims description 18
- 238000011161 development Methods 0.000 claims description 16
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 15
- 229960000583 acetic acid Drugs 0.000 claims description 15
- 239000012362 glacial acetic acid Substances 0.000 claims description 15
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 14
- 238000005259 measurement Methods 0.000 claims description 13
- 238000002156 mixing Methods 0.000 claims description 11
- 238000004809 thin layer chromatography Methods 0.000 claims description 11
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 10
- 229930182490 saponin Natural products 0.000 claims description 10
- 150000007949 saponins Chemical class 0.000 claims description 10
- 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 9
- 239000011248 coating agent Substances 0.000 claims description 9
- 238000000576 coating method Methods 0.000 claims description 9
- 235000019253 formic acid Nutrition 0.000 claims description 9
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 7
- -1 potassium ferricyanide Chemical compound 0.000 claims description 7
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 6
- 238000002137 ultrasound extraction Methods 0.000 claims description 6
- 230000005526 G1 to G0 transition Effects 0.000 claims description 5
- UFNDONGOJKNAES-UHFFFAOYSA-N Ginsenoside Rb1 Natural products CC(=CCCC(C)(OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CC(O)C45C)C UFNDONGOJKNAES-UHFFFAOYSA-N 0.000 claims description 5
- WVVOBOZHTQJXPB-UHFFFAOYSA-N N-anilino-N-nitronitramide Chemical compound [N+](=O)([O-])N(NC1=CC=CC=C1)[N+](=O)[O-] WVVOBOZHTQJXPB-UHFFFAOYSA-N 0.000 claims description 5
- 239000000945 filler Substances 0.000 claims description 5
- GZYPWOGIYAIIPV-JBDTYSNRSA-N ginsenoside Rb1 Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)O[C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(CC[C@H]4C(C)(C)[C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)CC[C@]4(C)[C@H]3C[C@H]2O)C)(C)CC1)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GZYPWOGIYAIIPV-JBDTYSNRSA-N 0.000 claims description 5
- TXEWRVNOAJOINC-UHFFFAOYSA-N ginsenoside Rb2 Natural products CC(=CCCC(OC1OC(COC2OCC(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CCC45C)C TXEWRVNOAJOINC-UHFFFAOYSA-N 0.000 claims description 5
- PWAOOJDMFUQOKB-WCZZMFLVSA-N ginsenoside Re Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](O[C@@H]2[C@H]3C(C)(C)[C@@H](O)CC[C@]3(C)[C@@H]3[C@@]([C@@]4(CC[C@@H]([C@H]4[C@H](O)C3)[C@](C)(CCC=C(C)C)O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C)(C)C2)O[C@H](CO)[C@@H](O)[C@@H]1O PWAOOJDMFUQOKB-WCZZMFLVSA-N 0.000 claims description 5
- AOGZLQUEBLOQCI-UHFFFAOYSA-N ginsenoside-Re Natural products CC1OC(OCC2OC(OC3CC4(C)C(CC(O)C5C(CCC45C)C(C)(CCC=C(C)C)OC6OC(CO)C(O)C(O)C6O)C7(C)CCC(O)C(C)(C)C37)C(O)C(O)C2O)C(O)C(O)C1O AOGZLQUEBLOQCI-UHFFFAOYSA-N 0.000 claims description 5
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 5
- 238000010992 reflux Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000000796 flavoring agent Substances 0.000 claims description 4
- 235000019634 flavors Nutrition 0.000 claims description 4
- 239000003205 fragrance Substances 0.000 claims description 4
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 3
- 239000012046 mixed solvent Substances 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 claims description 2
- 229910000387 ammonium dihydrogen phosphate Inorganic materials 0.000 claims description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 2
- 238000004811 liquid chromatography Methods 0.000 claims description 2
- 235000019837 monoammonium phosphate Nutrition 0.000 claims description 2
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 2
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 2
- 239000002023 wood Substances 0.000 claims description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 1
- 239000011574 phosphorus Substances 0.000 claims 1
- 229910052698 phosphorus Inorganic materials 0.000 claims 1
- 239000011591 potassium Substances 0.000 claims 1
- 229910052700 potassium Inorganic materials 0.000 claims 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical class [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims 1
- 239000002893 slag Substances 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 8
- 238000003908 quality control method Methods 0.000 abstract description 7
- 238000004458 analytical method Methods 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000000470 constituent Substances 0.000 abstract description 2
- 230000018109 developmental process Effects 0.000 description 13
- 208000032843 Hemorrhage Diseases 0.000 description 7
- 238000010521 absorption reaction Methods 0.000 description 7
- 239000002775 capsule Substances 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 239000003795 chemical substances by application Substances 0.000 description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 6
- 238000000605 extraction Methods 0.000 description 5
- 239000012467 final product Substances 0.000 description 5
- 238000004611 spectroscopical analysis Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 230000036407 pain Effects 0.000 description 4
- 239000006187 pill Substances 0.000 description 4
- 230000001737 promoting effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 206010046788 Uterine haemorrhage Diseases 0.000 description 3
- AOZUYISQWWJMJC-UHFFFAOYSA-N acetic acid;methanol;hydrate Chemical compound O.OC.CC(O)=O AOZUYISQWWJMJC-UHFFFAOYSA-N 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 229940000406 drug candidate Drugs 0.000 description 3
- KZUYUNXHKAGJQS-UHFFFAOYSA-N ethyl acetate;formic acid;propan-2-one Chemical compound OC=O.CC(C)=O.CCOC(C)=O KZUYUNXHKAGJQS-UHFFFAOYSA-N 0.000 description 3
- 239000003777 experimental drug Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 206010071321 Commotio retinae Diseases 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000023597 hemostasis Effects 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 238000005325 percolation Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 230000017423 tissue regeneration Effects 0.000 description 2
- 244000303512 Peucedanum ostruthium Species 0.000 description 1
- 235000007155 Peucedanum ostruthium Nutrition 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 229940126678 chinese medicines Drugs 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- GLMWWMZIEZJGMT-UHFFFAOYSA-N chloroform;formic acid;propan-2-one Chemical compound OC=O.CC(C)=O.ClC(Cl)Cl GLMWWMZIEZJGMT-UHFFFAOYSA-N 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000007941 film coated tablet Substances 0.000 description 1
- 239000007888 film coating Substances 0.000 description 1
- 238000009501 film coating Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 125000004836 hexamethylene group Chemical group [H]C([H])([*:2])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[*:1] 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- FBAFATDZDUQKNH-UHFFFAOYSA-M iron chloride Chemical compound [Cl-].[Fe] FBAFATDZDUQKNH-UHFFFAOYSA-M 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- 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
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
The invention discloses a kind of method of quality control of alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding, which includes the discrimination method and content assaying method of alizarin root of Dahurian angelica Chinese materia medica preparation；Identification be in alizarin root of Dahurian angelica Chinese materia medica preparation madder control medicinal material, rubimaillin, purpurin, Radix Notoginseng control medicinal material, notoginsenoside, ginsenoside, root of Dahurain angelica control medicinal material, Imperatorin, Isomperatorin, radix cyathulae control medicinal material, cyasterone indentification by TLC.The present invention is a kind of analysis method by controlling the active constituent control patent medicine quality of alizarin root of Dahurian angelica class Chinese materia medica preparation, to the quality standard of the alizarin root of Dahurian angelica carried out improve, it is perfect, on the basis of ministry standard, improve the quality control of drug, guarantee the stability and safety of its quality, it is horizontal to improve product technology, using the quality standard of modern analysis means General Promotion madder root of Dahurian angelica Chinese materia medica preparation, keeps the production technology of the alizarin root of Dahurian angelica more stringent reasonable.
Description
Technical field
The invention belongs to quality control technologies for traditional Chinese medicine fields, are related to a kind of alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding
Detection method.
Background technique
In existing technology, alizarin root of Dahurian angelica class Chinese materia medica preparation is mainly by radix cyathulae, Radix Notoginseng, madder, and the root of Dahurain angelica is made.Preparation method:
Radix cyathulae, the root of Dahurain angelica are ground into coarse powder, mix and stir with 70% ethyl alcohol of equivalent, bored profit, according to the percolation under liquid extract and extract item
(one annex of China's coastal port), using 10 times of 70% ethyl alcohol of amount as solvent soaking for 24 hours after slowly diacolation, liquid of filtering filtration returns
Ethyl alcohol is received, and being concentrated into relative density is about 1.10(35 DEG C), spray drying.Madder (system), Radix Notoginseng are ground into fine powder, and above-mentioned
Cream powder and starch merge, and mix, and 80% ethyl alcohol is added, mixes with proper auxiliary materials, and corresponding system is made according to conventional formulation method
Agent.Major function: promoting blood circulation and hemostasis, removing blood stasis for promoting tissue regeneration, swelling and pain relieving are used for caused by energy stagnation and blood stasis flooding, and the time prolongs
It is long, dripping more than, underbelly pain;The common above-mentioned disease person of commotio retinae uterine hemorrhage amount.
Alizarin root of Dahurian angelica class Chinese materia medica preparation proper mass standard only has a content and identifies, to the Indexes Comparison list of final product quality control
One.Preferably to control this quality, the ingredients of our Chinese medicines using in preparation carries out assay as Con trolling index, and to containing
Quantity measuring method carries out the investigation that the methods of linear relationship, stability, repeatability, rate of recovery are learned, the results showed that and this method is feasible,
Favorable reproducibility, specificity is strong, and controllability is good, can preferably control the inherent quality of this product, ensure that preparation process it is stable and
Quality it is controllable.
Summary of the invention
The object of the present invention is to provide the detection methods of alizarin root of Dahurian angelica class Chinese materia medica preparation, to solve existing alizarin root of Dahurian angelica Chinese materia medica preparation detection
The problems such as index components existing for method are single, characteristic differentiation is not comprehensive, assay specificity is poor, so as to more scientific
The quality for evaluating alizarin root of Dahurian angelica Chinese materia medica preparation, guarantees the stability and safety of its quality, improves product technology level, realizes using existing
For the quality standard of analysis means General Promotion madder root of Dahurian angelica class Chinese materia medica preparation, the gateway of a country is moved towards for the madder root of Dahurian angelica, better guarantee is provided.
To achieve the above object, present invention employs following technical solutions:
A kind of detection method of alizarin root of Dahurian angelica Chinese materia medica preparation that treating uterus bleeding, the detection method include alizarin root of Dahurian angelica Chinese medicine system
The discrimination method and content assaying method of agent；Identification is to madder, rubimaillin, the hydroxyl madder in alizarin root of Dahurian angelica Chinese materia medica preparation
Element, Radix Notoginseng, notoginsenoside, ginsenoside, the root of Dahurain angelica, Imperatorin, Isomperatorin, radix cyathulae, cyasterone thin-layer chromatography
Identify；The discrimination method are as follows:
Identify:
(1) method that madder is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking the madder for being equivalent to 15.25~20.55g of raw medicinal herbs
Root of Dahurian angelica Chinese materia medica preparation removes coating, finely ground, and 20~40ml of ether is added, and places 1 hour, and constantly shaking is extracted, and filtration, filtrate is waved
It goes, residue adds ethyl acetate 1ml to make to dissolve, spare as madder test solution；Separately take madder control medicinal material powder 0.8~
Control medicinal material solution is made with madder sample solution preparation method in 1.5g；It tests, draws above two molten according to thin-layered chromatography
Each 5 μ l of liquid puts the hexamethylene-the third for being 8~15: 4~10: 1~3: 4~10 with volume ratio on same silica gel g thin-layer plate respectively
Ketone-diethylamine-water be solvent expansion, take out, dry, set and inspected under 365nm ultraviolet lamp, in sample chromatogram, with compare
On the corresponding position of medicinal material chromatography, the spot of same color is shown；
Taking rubimaillin is reference substance, the solution that 1ml 0.1mg containing rubimaillin is made in methanol is added, as control
Product solution；It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of the madder test solution, put respectively in same
On silica gel g thin-layer plate, with volume ratio be 20~25: 3~6: 0.1~0.8 petroleum ether-ethyl acetate-glacial acetic acid be expansion
Agent expansion, takes out, dries, and spray is with dinitrophenylhydrazine ethyl alcohol test solution, in sample chromatogram, in position corresponding with reference substance chromatography
On, show the spot of same color；The petroleum ether that the step uses is 60~90 DEG C；
Taking purpurin is reference substance, methanol is added, 40 μ g of every 1ml hydroxyl rubican is made, molten as reference substance
Liquid；It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of the madder test solution, put respectively in same silica G
It is 15~20: 5~12: 1~2 ethyl acetate-acetones-formic acid as solvent using volume ratio on lamellae, is unfolded, takes out, dry in the air
It is dry, it sets and is inspected under 365nm ultraviolet lamp, in sample chromatogram, at the position corresponding to the chromatogram of the reference substance, show the spot of same color
Point；
(2) method that Radix Notoginseng is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking the alizarin root of Dahurian angelica for being equivalent to 13.6~18.6g of raw medicinal herbs
Chinese materia medica preparation removes coating, finely ground, and 15~30ml of methanol solution of 5% formic acid is added, is heated to reflux 30 minutes, lets cool, filters,
Filtrate is flung to, and residue adds methanol 2ml to make to dissolve, spare as Radix Notoginseng test solution；Separately take Radix Notoginseng control medicinal material powder 0.8~
Control medicinal material solution is made with Radix Notoginseng sample solution preparation method in 1.5g；It tests, draws above two molten according to thin-layered chromatography
Each 5 μ l of liquid puts respectively on same silica gel g thin-layer plate, is for 8~12: 6~9: 1~3 benzene-acetic ether-methanoic acid with volume ratio
Solvent is unfolded, and takes out, dries, set and inspect under 365nm ultraviolet lamp, in sample chromatogram, corresponding with reference medicine chromatography
On position, the spot of same color is shown；
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Reference substance adds methanol that solution of every 1ml containing 1mg is made, as reference substance
Solution；It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of Radix Notoginseng test solution, put respectively thin in same silica G
On laminate, using the upper solution that volume ratio is 5~12: 3~7: 0.5~1.5 chloroform-methanols-glacial acetic acid as solvent, expansion,
It takes out, dries, spray with ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C；In sample chromatogram, with reference substance
On the corresponding position of chromatography, the spot of same color is shown；
Ginsenoside Rb1's reference substance and ginsenoside Re's reference substance are taken, adds methanol that mixing of every 1ml containing 0.5mg is made molten
Liquid, as reference substance solution；It is tested according to thin-layered chromatography, draws reference substance solution and each 3 μ l of Radix Notoginseng test solution point respectively
In on same silica gel g thin-layer plate, with volume ratio for 10~18: 35~45: 17~25: 8~12 chloroforms-ethyl acetate-first
10 DEG C of alcohol-water lower layer's solution arranged below are solvent, are unfolded, and take out, dry, and spray with sulfuric acid solution, are heated at 105 DEG C
Spot development is clear；In sample chromatogram, at the position corresponding to the chromatogram of the reference substance, identical colored speckles are shown；
(3) method that the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking the alizarin root of Dahurian angelica for being equivalent to 14.6~19.8g of raw medicinal herbs
Chinese materia medica preparation removes coating, finely ground, adds 10~40ml of petroleum ether, and shaking is extracted 30 minutes, and filtration, filter residue is spare, and filtrate is flung to
Petroleum ether is spare as root of Dahurain angelica test solution to 2ml；The root of Dahurain angelica 0.4~0.9g of control medicinal material separately is taken, adds petroleum ether (60~90
DEG C) 5~10ml, shaking extraction 30 minutes, standing, supernatant is as control medicinal material solution, according to thin-layered chromatography test, absorption
Above two each 5 μ l of solution is put respectively on same silica gel g thin-layer plate, with volume ratio for 12~20: 2~5: 0.1~0.8 stone
Oily ether-ethyl acetate-glacial acetic acid is solvent, is unfolded at 25 DEG C or less, takes out, dries, set and inspect under 365nm ultraviolet lamp；For
In test product chromatography, on position corresponding with reference medicine chromatography, the spot of same color is shown；The petroleum ether that the step uses is
60~90 DEG C；
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added, the mixing that every 1ml contains 1mg is made
Solution, as reference substance solution；It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of root of Dahurain angelica test solution, respectively
Point is 5~9: 2~5: 0~1 petroleum ether-ether-formic acid as solvent using volume ratio, 25 on same silica gel g thin-layer plate
DEG C or less expansion, take out, dry, set and inspected under ultraviolet lamp；In sample chromatogram, in position corresponding with reference substance chromatography
On, show the spot of same color；The petroleum ether that the step uses is 30~60 DEG C；
(4) method that radix cyathulae is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking the madder for being equivalent to 12.7~18.9g of raw medicinal herbs
Root of Dahurian angelica Chinese materia medica preparation, adds 30~50ml of ethyl alcohol, is ultrasonically treated 30 minutes, and filtration, filtrate is evaporated, and residue adds diethyl ether 15~25ml, vibration
Extraction is shaken, is volatilized, residue adds ethyl acetate 1ml to make to dissolve, as radix cyathulae test solution；Separately take radix cyathulae control medicinal material 1
~4g adds 20~40ml of ethyl alcohol, and control medicinal material solution is made with radix cyathulae sample solution preparation method；It is tried according to thin-layer chromatography
It tests, draws 6~8 μ l of radix cyathulae test solution, 3~5 μ l of control medicinal material solution, put respectively on same silica G lamellae,
Using volume ratio for 10~25: 8~18: 1~3 toluene: ethyl acetate: formic acid is unfolded as solvent, takes out, dries, and sprays with body
Product is than for 1: 1% ferric trichloride: it is clear that the mixed solution of 1% potassium ferricyanide is heated to spot development；In sample chromatogram, with
On the corresponding position of reference medicine chromatography, the spot of same color is shown；
Cyasterone reference substance is taken, adds methanol that solution of every 1ml containing 0.5mg is made, as reference substance solution；According to thin layer color
Spectrometry test, draws 5~10 μ l of radix cyathulae test solution and 5 μ l of reference substance solution, is put respectively in same silica gel g thin-layer plate
On, it is 13~18: 3~5: 1~2 chloroforms-acetone-formic acid as solvent using volume ratio, is unfolded, take out, dry, sprays with 10% sulphur
Sour ethanol solution, it is clear to be heated to spot development at 105 DEG C, sets and inspects under ultraviolet lamp；In sample chromatogram, with reference substance
On the corresponding position of chromatography, the spot of same color is shown；
The content assaying method of the madder root of Dahurian angelica Chinese materia medica preparation, including using in high effective liquid chromatography for measuring madder root of Dahurian angelica Chinese materia medica preparation
The amount of Imperatorin contained by the root of Dahurain angelica；
Chromatographic condition and system suitability:
Flow stationary phase: selecting octadecylsilane chemically bonded silica is the chromatographic column of filler；
Mobile phase: made with the mixed solvent that component A and component B that volume ratio is (30: 70)~(65: 35) are constituted
For mobile phase；The component A is the acetonitrile and methanol solution that volume ratio is 5: 1, and component B is 0.01~0.05mol/L
Potassium dihydrogen phosphate or sodium dihydrogen phosphate or ammonium dihydrogen phosphate, every 1000ml flowing are added to the triethylamine of 3ml,
PH value is adjusted to 3.0~4.5 with phosphoric acid solution；
Detection wavelength: 275~315nm；
Column temperature: 25~45 DEG C；
Flow velocity: 0.8~1.2ml/min of flow velocity；
The preparation of test solution: taking the alizarin root of Dahurian angelica Chinese materia medica preparation for being equivalent to raw medicinal herbs 17.56g, accurately weighed, finely ground, takes
1.8g, it is accurately weighed, it sets in stuffed conical flask, 50% methanol 50ml, weighed weight, ultrasonic extraction 30~40 is added in precision
Minute, it lets cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filter, discard primary filtrate, remaining filtrate through hole
Diameter is 0.45 μm of miillpore filter filtration to get test solution；
The preparation of reference substance solution: precision weighs Imperatorin reference substance, accurately weighed, adds methanol that every 1ml is made and contains
The solution of 20 μ g, shake up to get；
Measuring method: accurate absorption reference substance solution and each 10 μ l of test solution respectively, injection liquid chromatograph, measurement,
To obtain the final product.
The present invention is a kind of analysis side by controlling the active constituent control patent medicine quality of alizarin root of Dahurian angelica class Chinese materia medica preparation
Method, to the quality standard of the alizarin root of Dahurian angelica carried out improve, it is perfect, on the basis of ministry standard, improve drug quality control, guarantee
The stability and safety of its quality improve product technology level, utilize modern analysis means General Promotion madder root of Dahurian angelica Chinese materia medica preparation
Quality standard, make the production technology of the alizarin root of Dahurian angelica more it is stringent rationally.
Present invention understands that index components existing for certainly existing alizarin root of Dahurian angelica Chinese materia medica preparation control method are single, characteristic differentiation is not complete
The problems such as face, poor assay specificity, guarantees its quality so as to the quality of more scientific evaluation madder root of Dahurian angelica Chinese materia medica preparation
Stability and safety improve product technology level, realize and utilize modern analysis means General Promotion madder root of Dahurian angelica class Chinese materia medica preparation
Quality standard moves towards the gateway of a country for the madder root of Dahurian angelica and provides better guarantee.
The present invention has done following research to the content of alizarin root of Dahurian angelica Chinese materia medica preparation Imperatorin:
One, experimental drug
1. the prescription of experimental drug: radix cyathulae 600g Radix Notoginseng 85g madder (system) 85g root of Dahurain angelica 85g
2. the preparation method of experimental drug: above four taste, radix cyathulae, the root of Dahurain angelica are ground into coarse powder, mix and stir with 70% ethyl alcohol of equivalent, is bored
Profit, according to the percolation under liquid extract and extract item, (one annex of China's coastal port is with 10 times of 70% ethyl alcohol of amount
Solvent soaking for 24 hours after slowly diacolation, liquid of filtering filtration recycles ethyl alcohol, and being concentrated into relative density is about 1.10(35 DEG C), it is spraying dry
It is dry.Madder (system), Radix Notoginseng are ground into fine powder, merge with above-mentioned cream powder and starch, mix, and 80% alcohol granulation is added, dry, add
Magnesium stearate mixes, and arranges, is pressed into 1000, film coating to get.
Two, research process:
1, instrument, reagent, sample: Waters515 type high performance liquid chromatograph, Waters2487UV-Vis detector,
WDL-95 chromatographic work station, UV-1800 type ultraviolet-uisible spectrophotometer, 800 type settling vessels (Shanghai Surgical Operation Equipment Factory), QX type
Supersonic oscillations instrument.Reagent: acetonitrile, methanol, potassium dihydrogen phosphate, triethylamine are chromatographically pure, and water is double distilled water.Imperatorin pair
According to product (Chinese pharmaceutical biological product inspection institute).Alizarin root of Dahurian angelica piece (Gansu Lanyao Pharmaceuticals Co., Ltd.).
2, it measures the selection of wavelength: taking Imperatorin reference substance solution, with ultraviolet-uisible spectrophotometer, in wavelength model
It encloses and is scanned measurement in 200~400nm wave-length coverage.The result shows that: wavelength has preferably in 275nm~315nm range
Effect, Imperatorin maximum absorption wavelength are 300nm.
3, chromatographic condition: chromatographic column: μ-Bondapak2 C18(4.6nm × 250nm, 5 μm, Chinese Academy of Sciences's Dalian Chemical Physics
Chromatogram research analysis center of country of research institute)；Mobile phase: the acetonitrile of volume ratio 45: 55: methanol (5: 1) -0.02mol/
L potassium dihydrogen phosphate mixed solvent adjusts pH with phosphoric acid solution as mobile phase (triethylamine that 3ml is added in every 1000ml)
It is worth to 4.0.Flow velocity: 1.0ml/min.Column temperature: 30 DEG C of sample volumes: 5 μ L.
4, the preparation of test solution:
(1) determination of extraction time:
Alizarin root of Dahurian angelica Chinese materia medica preparation is taken, it is finely ground, about 1.5g is taken, it is four parts, accurately weighed, it sets in stuffed conical flask, precision plus 50%
Methanol 25ml, weighed weight are ultrasonically treated (power 250W, frequency 40kHz) 20,30,40,50 minutes respectively, let cool, then claim
Determine weight, the weight of less loss is supplied with 50% methanol, is shaken up, filters.Subsequent filtrate is filtered with (0.45 μm) of miillpore filter, measurement.
It the results are shown in Table 1.
According to the result of table 1 it is found that the content for extracting 40 minutes and 50 minutes Imperatorins is close, therefore select 40 minutes ultrasonic
As extraction time.
(2) this product for being equivalent to raw medicinal herbs 17.56g is taken, it is accurately weighed, it is finely ground, about 1.8g is taken, it is accurately weighed, set tool plug
In conical flask, 50% methanol 50ml is added in precision, and weighed weight ultrasonic extraction 40 minutes, is let cool, then weighed weight, with 50%
Methanol supplies the weight of less loss, filtration, discards primary filtrate, and remaining filtrate is 0.45 μm of miillpore filter filtration by aperture, i.e.,
Obtain test solution.
5, the preparation of reference substance solution: precision weighs Imperatorin reference substance, accurately weighed, adds methanol that every 1ml is made and contains
Have the solution of 20 μ g Imperatorins, shake up to get.
6, by prescription (removing the root of Dahurain angelica), pharmaceutical preparation the preparation of negative test solution: is prepared into according to preparation process.It presses
According to the preparation method of test solution, negative test solution is similarly made.It is analyzed according to content determination, blank sample chromatography
Figure does not interfere with peak in Imperatorin peak retention time.The result shows that negative noiseless.
7, the investigation of linear relationship
Precision draws Imperatorin reference substance solution (20 μ g/ml) 8,10,12,16,20,24 μ l, by above-mentioned chromatostrip
Part sample introduction measures peak area, the results are shown in Table 2.
Using respective sample volume as abscissa, peak area is absorbed as ordinate using reference substance, standard curve is drawn, obtains standard
Curvilinear equation Y=5937.0X+98.61(r=0.9998), make Fig. 1 to absorb peak area value to sample volume (μ g), obtains a straight line.
The above result shows that Imperatorin within the scope of 0.16~0.48 μ g, absorbs peak area value and sample volume in good linear
Relationship.
8, precision test precision absorption Imperatorin reference substance solution 20ul, continuous sample introduction 5 times, with reference substance peak face
Product is inspection target, and as a result the RSD of Imperatorin peak area is 1.10%, shows that the precision of this method is good.
9, stability test precision draws above-mentioned test solution 20ul, respectively in 0h, 4h, 8h, 12h, 20h and for 24 hours into
Sample, with test sample calculated by peak area RSD for 1.12%.Measurement result shows Imperatorin interior for 24 hours basicly stable.
10, repetitive test precision weigh same lot number alizarin root of Dahurian angelica piece fine powder it is appropriate, 5 parts are done altogether, by test solution
Preparation carries out time-and-motion study with the method under measurement item.
Measuring content average value as shown in Table 3 is 0.0762mg/ piece, and RSD 1.92% illustrates that this method measures
It is reproducible.
11, recovery test precision weighs the sample about 8g of known content, 6 parts, accurately weighed, is respectively placed in tool plug cone
In shape bottle, precision, which weighs addition Imperatorin reference substance, respectively the results are shown in Table 4 according to the content assaying method of sample.
The result shows that this method measurement Imperatorin shows that sample recovery rate is good.
12, the measurement of sample size takes each 3 parts of 3 batches of samples, measures every gram of raw medicinal herbs Imperatoria ostruthium in sample according to the above method
The content of element, measurement result are shown in Table 5.
By the test result of table 5 it is found that the average content value of three batches of sample Imperatorins is 0.076mg, RSD is
4.2%.In conjunction with production actual state and international food Drug Administration standard, therefore require to be equivalent in the sheet of 1g raw medicinal herbs
Medicine preparation is containing the root of Dahurain angelica with Imperatorin (C18H14O4Meter) meter, 0.067mg must not be less than.
Detailed description of the invention
Fig. 1 is Imperatorin linear relationship measurement result figure of the present invention.
Specific embodiment
The present invention is further illustrated below by specific embodiment.
A kind of alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding of the present invention mainly by radix cyathulae, Radix Notoginseng, madder,
Root of Dahurain angelica composition is made various according to the conventional method of pharmacy and is suitable for the acceptable capsule of clinical treatment, tablet, particle
Agent, pill and pill etc..By promoting blood circulation and hemostasis, removing blood stasis for promoting tissue regeneration, swelling and pain relieving, it to be used for caused by energy stagnation and blood stasis uterine hemorrhage mistake
More, the time extends, dripping more than, underbelly pain；The common above-mentioned disease person of commotio retinae uterine hemorrhage amount.The provided system
The method of quality control of agent is stable, reliable, is better than ministry standard, more effectively ensure that the quality of alizarin root of Dahurian angelica class Chinese materia medica preparation.Its
Method of quality control includes the discrimination method and content assaying method of alizarin root of Dahurian angelica Chinese materia medica preparation.
Embodiment 1 is a kind of detection side of alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding of capsule for dosage form
Method, the discrimination method are as follows:
Character: removing capsule shells, and alizarin root of Dahurian angelica Chinese materia medica preparation content is the powder of brown；Gas fragrance, acrid flavour, slight bitter.
Identify:
(1) method that madder is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking 20 this product, removes capsule shells, ether is added
20ml is placed 1 hour, and constantly shaking is extracted, and filtration, filtrate is flung to, and residue adds ethyl acetate 1ml to make to dissolve, as test sample
Solution.Madder control medicinal material powder 0.8g separately is taken, control drug solution is made with sample solution preparation method.According to thin-layer chromatography
Method test, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively on same silica gel g thin-layer plate, with
Cyclohexane-acetone-diethylamine-water (8: 4: 1: 5) is solvent, is unfolded, and takes out, dries, set and examine under ultraviolet lamp (365nm)
Depending on position corresponding with reference medicine chromatography, showing the spot of same color in sample chromatogram.
Taking rubimaillin is reference substance, the solution that 1ml 0.1mg containing rubimaillin is made in methanol is added, as control
Product solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of test solution, put respectively in same silica G thin layer
On plate, with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (20: 3: 0.2) for solvent, it is unfolded, takes out, dry in the air
Dry, spray in sample chromatogram, on position corresponding with reference medicine chromatography, shows identical face with dinitrophenylhydrazine ethyl alcohol test solution
The spot of color.
Taking purpurin is reference substance, methanol is added, 40 μ g of every 1ml hydroxyl rubican is made, molten as reference substance
Liquid.It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of test solution, put respectively in same silica gel g thin-layer plate
On, with ethyl acetate-acetone-formic acid (15: 7: 1) for solvent, it is unfolded, takes out, dry, set ultraviolet lamp (365nm)
Under inspect, in sample chromatogram, on position corresponding with reference medicine chromatography, show same color spot.
(2) method that Radix Notoginseng is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking 17 this product, removes capsule shells, 5% first is added
The methanol solution 20ml of acid, is heated to reflux 30 minutes, lets cool, and filters, and filtrate is flung to, and residue adds methanol 2ml to make to dissolve, as confession
Test sample solution.Radix Notoginseng control medicinal material powder 0.8g separately is taken, control drug solution is made with sample solution preparation method.According to thin layer
Chromatography experiment, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively in same silica gel g thin-layer plate
On, with benzene-acetic ether-methanoic acid (8: 7: 1) for solvent, it is unfolded, takes out, dry, set and inspected under ultraviolet lamp (365nm),
In sample chromatogram, on position corresponding with reference medicine chromatography, the spot of same color is shown.
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Reference substance adds methanol that solution of every 1ml containing 1mg is made, as reference substance
Solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of test solution, put respectively in same silica gel g thin-layer plate
On, using chloroform-methanol-water-glacial acetic acid (7: 2: 4: 1) upper solution as solvent, it is unfolded, takes out, dry, sprays with sulfuric acid second
It is clear to be heated to spot development at 105 DEG C for alcoholic solution (1 → 10).In sample chromatogram, in position corresponding with reference substance chromatography
On, show the spot of same color.
Ginsenoside Rb1's reference substance and ginsenoside Re's reference substance are taken, adds methanol that mixing of every 1ml containing 0.5mg is made molten
Liquid, as reference substance solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 3 μ l of test solution.Put respectively in
It is molten with chloroform -10 DEG C of lower layers arranged below of acetate-methanol-water (10: 35: 17: 8) on same silica gel g thin-layer plate
Liquid is solvent, is unfolded, and takes out, dries, and sprays with sulfuric acid solution (1 → 10), it is clear to be heated to spot development at 105 DEG C.For examination
In product chromatography, at the position corresponding to the chromatogram of the reference substance, identical colored speckles are shown.
(3) method that the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking 18 this product, removes capsule shells, add petroleum ether
(60~90 DEG C) 20ml, shaking are extracted 30 minutes, and filtration, filter residue is spare, and filtrate flings to petroleum ether to about 2ml, as test sample
Solution.Root of Dahurain angelica control medicinal material 0.4g separately is taken, adds petroleum ether (60~90 DEG C) 5ml, shaking is extracted 30 minutes, is stood, and supernatant is made
For control medicinal material solution.According to thin-layered chromatography test, draw each 5 μ l(test solution of above two solution give over to it is spare), point
Other point is on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (12: 2: 0.3) for solvent,
It is unfolded at 25 DEG C or less, takes out, dry, set and inspected under ultraviolet lamp (365nm).In sample chromatogram, with reference medicine chromatography
On corresponding position, the spot of same color is shown.
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added, the mixing that every 1ml contains 1mg is made
Solution, as reference substance solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of three kinds of solution of test solution,
It is put respectively on same silica gel g thin-layer plate, with petroleum ether (30~60 DEG C)-ether (5: 2) for solvent, is opened up at 25 DEG C or less
It opens, takes out, dry, set and inspected under ultraviolet lamp (365nm).In sample chromatogram, in position corresponding with reference medicine chromatography
On, show the spot of same color.
(4) method that radix cyathulae is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking 14 this product, removes capsule shells, add ethyl alcohol
30ml is ultrasonically treated 30 minutes, and filtration, filtrate is evaporated, and residue adds diethyl ether 15ml, and shaking is extracted, volatilized, residue adds ethyl acetate
1ml makes to dissolve, as test solution；Radix cyathulae control medicinal material 2g separately is taken, adds ethyl alcohol 20ml, sample solution preparation method system
Drug solution is shone in pairs.According to TLC test, 6 μ l of test solution, 3 μ l of control medicinal material solution are drawn, is put respectively in same
On silica G lamellae, with toluene: ethyl acetate: formic acid (10: 9: 1) is solvent, is unfolded, and takes out, dries, and is sprayed with 1% 3
Iron chloride: it is clear that the mixed solution of 1% potassium ferricyanide (1: 1) is heated to spot development.In sample chromatogram, with comparison medicine
Wood color is composed on corresponding position, and the spot of same color is shown.
Cyasterone reference substance is taken, adds methanol that solution of every 1ml containing 0.5mg is made, as reference substance solution.According to thin layer color
Spectrometry test, draws 5 μ l of test solution and 5 μ l of reference substance solution, is put respectively on same silica gel g thin-layer plate, with chloroform-the third
Ketone-formic acid (13: 3: 1) is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, is heated to spot at 105 DEG C
Colour developing is clear, sets and inspects under ultraviolet lamp (365nm).In sample chromatogram, on position corresponding with reference medicine chromatography, show
The spot of same color.
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: selecting octadecylsilane chemically bonded silica is the chromatographic column of filler；
Mobile phase: the acetonitrile of volume ratio 45: 55: methanol (5: 1) -0.02mol/l potassium dihydrogen phosphate is mixed
Bonding solvent adjusts pH value to 4.0 as mobile phase (triethylamine that 3ml is added in every 1000ml), with phosphoric acid solution；
Detection wavelength: 300nm；
Column temperature: 30 DEG C；
Flow velocity: 1.0ml/min.
The preparation of reference substance solution: precision weighs Imperatorin reference substance, accurately weighed, adds methanol that every 1ml is made and contains
The solution of 20 μ g, shake up to get.
The preparation of test solution: taking this product for being equivalent to raw medicinal herbs 17.56g, accurately weighed, finely ground, takes about 1.8g,
It is accurately weighed, it sets in stuffed conical flask, the methanol 25ml of precision addition 50%, weighed weight, ultrasonic extraction 30~40 minutes,
It lets cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filter, discard primary filtrate, remaining filtrate is by aperture
0.45 μm of miillpore filter filtration is to get test solution.
Measuring method: accurate absorption reference substance solution and each 10 μ l of test solution respectively, injection liquid chromatograph, measurement,
To obtain the final product.
The Chinese medicine preparation for being equivalent to 1g raw medicinal herbs contains the root of Dahurain angelica with Imperatorin (C18H14O4Meter) meter, it must not be less than
0.067mg。
Embodiment 2 is a kind of detection method of alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding of tablet for dosage form,
The discrimination method are as follows:
Character: this product is Film coated tablets, is removed brown after being coated: gas fragrance, acrid flavour, slight bitter.
Identify:
(1) method that madder is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: 25 this product taken, coating is removed, it is finely ground, and second is added
Ether 40ml is placed 1 hour, and constantly shaking is extracted, and filtration, filtrate is flung to, and residue adds ethyl acetate 1ml to make to dissolve, as examination
Product solution.Madder control medicinal material powder 1.5g separately is taken, control drug solution is made with sample solution preparation method.According to thin layer color
Spectrometry test, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively on same silica gel g thin-layer plate,
With cyclohexane-acetone-diethylamine-water (15: 10: 3: 10) for solvent, it is unfolded, takes out, dry, set and inspected under ultraviolet lamp,
In sample chromatogram, on position corresponding with reference medicine chromatography, the spot of same color is shown.
Taking rubimaillin is reference substance, the solution that 1ml 0.1mg containing rubimaillin is made in methanol is added, as control
Product solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of test solution, put respectively in same silica G thin layer
On plate, with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (25: 5: 0.8) for solvent, it is unfolded, takes out, dry,
Spray is with dinitrophenylhydrazine ethyl alcohol test solution, in sample chromatogram, on position corresponding with reference medicine chromatography, shows same color
Spot.
Taking purpurin is reference substance, methanol is added, 40 μ g of every 1ml hydroxyl rubican is made, molten as reference substance
Liquid.It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of test solution, put respectively in same silica gel g thin-layer plate
On, with ethyl acetate-acetone-formic acid (20: 11: 2) for solvent, it is unfolded, takes out, dry, set and inspected under ultraviolet lamp,
In sample chromatogram, on position corresponding with reference medicine chromatography, the spot of same color is shown.
(2) method that Radix Notoginseng is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: 22 this product taken, coating is removed, it is finely ground, and it is added 5%
The methanol solution 30ml of formic acid is heated to reflux 30 minutes, lets cool, and filtration, filtrate is flung to, and residue adds methanol 2ml to make to dissolve, as
Test solution.Radix Notoginseng control medicinal material powder 1.5g separately is taken, control drug solution is made with sample solution preparation method.According to thin
Layer chromatography test, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively in same silica G thin layer
On plate, with benzene-acetic ether-methanoic acid (12: 9: 2.5) for solvent, it is unfolded, takes out, dry, set and inspected under ultraviolet lamp, supply
In test product chromatography, on position corresponding with reference medicine chromatography, the spot of same color is shown.
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Reference substance adds methanol that solution of every 1ml containing 1mg is made, as reference substance
Solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of test solution, put respectively in same silica gel g thin-layer plate
On, using chloroform-methanol-water-glacial acetic acid (12: 7: 5: 1) upper solution as solvent, it is unfolded, takes out, dry, sprays with sulfuric acid
It is clear to be heated to spot development at 105 DEG C for ethanol solution (1 → 10).In sample chromatogram, in position corresponding with reference substance chromatography
It sets, shows the spot of same color.
Ginsenoside Rb1's reference substance and ginsenoside Re's reference substance are taken, adds methanol that mixing of every 1ml containing 0.5mg is made molten
Liquid, as reference substance solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 3 μ l of test solution.It puts respectively in same
It is molten with chloroform -10 DEG C of lower layers arranged below of acetate-methanol-water (18: 45: 25: 12) on one silica gel g thin-layer plate
Liquid is solvent, is unfolded, and takes out, dries, and sprays with sulfuric acid solution, it is clear to be heated to spot development at 105 DEG C.Sample chromatogram
In, at the position corresponding to the chromatogram of the reference substance, show identical colored speckles.
(3) method that the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: 22 this product taken, coating is removed, it is finely ground, and add petroleum
Ether (60~90 DEG C) 40ml, shaking are extracted 30 minutes, and filtration, filter residue is spare, and filtrate flings to petroleum ether to about 2ml, as examination
Product solution.Root of Dahurain angelica control medicinal material 0.9g separately is taken, adds petroleum ether (60~90 DEG C) 10ml, shaking is extracted 30 minutes, is stood, supernatant
As control medicinal material solution.According to thin-layered chromatography test, draw each 5 μ l(test solution of above two solution give over to it is spare),
It is put respectively on same silica gel g thin-layer plate, is expansion with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (20: 5: 0.8)
Agent is unfolded at 25 DEG C or less, takes out, dries, set and inspect under ultraviolet lamp.In sample chromatogram, corresponding to reference medicine chromatography
Position on, show same color spot.
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added, the mixing that every 1ml contains 1mg is made
Solution, as reference substance solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of three kinds of solution of test solution,
It is put respectively on same silica gel g thin-layer plate, with petroleum ether (30~60 DEG C)-ether-formic acid (9: 5: 1) for solvent, at 25 DEG C
It is unfolded below, takes out, dry, set and inspected under ultraviolet lamp.In sample chromatogram, in position corresponding with reference medicine chromatography
On, show the spot of same color.
(4) method that radix cyathulae is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking and is equivalent to 20 this product, adds ethyl alcohol
50ml is ultrasonically treated 30 minutes, and filtration, filtrate is evaporated, and residue adds diethyl ether 25ml, and shaking is extracted, volatilized, residue adds ethyl acetate
1ml makes to dissolve, as test solution；Radix cyathulae control medicinal material 4g separately is taken, adds ethyl alcohol 40ml, sample solution preparation method
Control drug solution is made.According to TLC test, 8 μ l of test solution, 5 μ l of control medicinal material solution are drawn, is put respectively in same
On one silica G lamellae, with toluene: ethyl acetate: formic acid (25: 18: 3) be solvent, be unfolded, take out, dry, spray with
1% ferric trichloride: it is clear that the mixed solution of 1% potassium ferricyanide (1: 1) is heated to spot development.In sample chromatogram, with
On the corresponding position of reference medicine chromatography, the spot of same color is shown.
Cyasterone reference substance is taken, adds methanol that solution of every 1ml containing 0.5mg is made, as reference substance solution.According to thin layer color
Spectrometry test, draws 10 μ l of test solution and 5 μ l of reference substance solution, is put respectively on same silica gel g thin-layer plate, with chloroform-
Acetone-formic acid (18: 5: 2) is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, is heated to spot at 105 DEG C
Point colour developing is clear, sets and inspects under ultraviolet lamp.In sample chromatogram, on position corresponding with reference medicine chromatography, show identical
The spot of color.
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: selecting octadecylsilane chemically bonded silica is the chromatographic column of filler；
Mobile phase: the acetonitrile of volume ratio 45: 55: methanol (5: 1) -0.02mol/l potassium dihydrogen phosphate mixing
Solvent adjusts pH value to 4.0 as mobile phase (triethylamine that 3ml is added in every 1000ml), with phosphoric acid solution；
Detection wavelength: 300nm；
Column temperature: 30 DEG C；
Flow velocity: 1.0ml/min.
The preparation of test solution: taking this product for being equivalent to raw medicinal herbs 17.56g, accurately weighed, finely ground, takes about 1.8g,
It is accurately weighed, it sets in stuffed conical flask, the methanol 25ml of precision addition 50%, weighed weight, ultrasonic extraction 30~40 minutes,
It lets cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filter, discard primary filtrate, remaining filtrate is by aperture
0.45 μm of miillpore filter filtration is to get test solution.
The preparation of reference substance solution: precision weighs Imperatorin reference substance, accurately weighed, adds methanol that every 1ml is made and contains
The solution of 20 μ g, shake up to get.
Measuring method: accurate absorption reference substance solution and each 10 μ l of test solution respectively, injection liquid chromatograph, measurement,
To obtain the final product.
The Chinese medicine preparation for being equivalent to 1g raw medicinal herbs contains the root of Dahurain angelica with Imperatorin (C18H14O4Meter) meter, it must not be less than
0.067mg。
Embodiment 3 is a kind of detection method of alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding of pill for dosage form,
The discrimination method are as follows:
Character: alizarin root of Dahurian angelica pill is brown: gas fragrance, acrid flavour, slight bitter.
Identify:
(1) method that madder is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: this product of 23 balls is taken, it is finely ground, and ether 30ml is added,
It places 1 hour, constantly shaking is extracted, and filtration, filtrate is flung to, and residue adds ethyl acetate 1ml to make to dissolve, as test solution.
Madder control medicinal material powder 1.2g separately is taken, control drug solution is made with sample solution preparation method.It is tried according to thin-layered chromatography
Test, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively on same silica gel g thin-layer plate, with hexamethylene
Alkane-acetone-diethylamine-water (12: 5: 2: 5) is solvent, is unfolded, and takes out, dries, set and inspect under ultraviolet lamp, test sample color
In spectrum, on position corresponding with reference medicine chromatography, the spot of same color is shown.
Taking rubimaillin is reference substance, the solution that 1ml 0.1mg containing rubimaillin is made in methanol is added, as control
Product solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of test solution, put respectively in same silica G thin layer
On plate, with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (22: 4: 0.5) for solvent, it is unfolded, takes out, dry in the air
Dry, spray in sample chromatogram, on position corresponding with reference medicine chromatography, shows identical face with dinitrophenylhydrazine ethyl alcohol test solution
The spot of color.
Taking purpurin is reference substance, methanol is added, 40 μ g of every 1ml hydroxyl rubican is made, molten as reference substance
Liquid.It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of test solution, put respectively in same silica gel g thin-layer plate
On, with ethyl acetate-acetone-formic acid (17: 9: 1) for solvent, it is unfolded, takes out, dry, set and inspected under ultraviolet lamp, supplies
In test product chromatography, on position corresponding with reference medicine chromatography, the spot of same color is shown.
(2) method that Radix Notoginseng is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: this product of 20 balls is taken, it is finely ground, and 5% formic acid is added
Methanol solution 25ml is heated to reflux 30 minutes, lets cool, and filtration, filtrate is flung to, and residue adds methanol 2ml to make to dissolve, as test sample
Solution.Radix Notoginseng control medicinal material powder 1.1g separately is taken, control drug solution is made with sample solution preparation method.According to thin-layer chromatography
Method test, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively on same silica gel g thin-layer plate, with
Benzene-acetic ether-methanoic acid (10: 7: 2) is solvent, is unfolded, and takes out, dries, set and inspect under ultraviolet lamp, sample chromatogram
In, on position corresponding with reference medicine chromatography, show the spot of same color.
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Reference substance adds methanol that solution of every 1ml containing 1mg is made, as reference substance
Solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of test solution, put respectively in same silica gel g thin-layer plate
On, using the upper solution of chloroform-methanol-glacial acetic acid (9: 5: 1) as solvent, it is unfolded, takes out, dry, sprays molten with sulfuric acid ethyl alcohol
It is clear to be heated to spot development at 105 DEG C for liquid.In sample chromatogram, on position corresponding with reference substance chromatography, identical face is shown
The spot of color.
Ginsenoside Rb1's reference substance and ginsenoside Re's reference substance are taken, adds methanol that mixing of every 1ml containing 0.5mg is made molten
Liquid, as reference substance solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 3 μ l of test solution.It puts respectively in same
On one silica gel g thin-layer plate, with chloroform -10 DEG C of acetate-methanol-water (14: 40: 22: 9) lower layer's solution arranged below
For solvent, it is unfolded, takes out, dry, sprays with sulfuric acid solution, it is clear to be heated to spot development at 105 DEG C.In sample chromatogram,
At the position corresponding to the chromatogram of the reference substance, identical colored speckles are shown.
(3) method that the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: 21 ball this product are taken, it is finely ground, and add petroleum ether (60~90
DEG C) 25ml, shaking extraction 30 minutes, filtration, filter residue is spare, and filtrate flings to petroleum ether to about 2ml, as test solution.Separately
Root of Dahurain angelica control medicinal material 0.6g is taken, petroleum ether (60~90 DEG C) 8ml is added, shaking is extracted 30 minutes, is stood, supernatant is as comparison medicine
Material solution.According to thin-layered chromatography test, draw each 5 μ l(test solution of above two solution give over to it is spare), put respectively in same
On one silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (15: 4: 0.5) for solvent, at 25 DEG C
It is unfolded below, takes out, dry, set and inspected under ultraviolet lamp.In sample chromatogram, on position corresponding with reference medicine chromatography,
The spot of aobvious same color.
Imperatorin reference substance and Isomperatorin reference substance are taken, ethyl acetate is added, the mixing that every 1ml contains 1mg is made
Solution, as reference substance solution.It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of three kinds of solution of test solution,
It is put respectively on same silica gel g thin-layer plate, with petroleum ether (30~60 DEG C)-ether (7: 4) for solvent, is opened up at 25 DEG C or less
It opens, takes out, dry, set and inspected under ultraviolet lamp.In sample chromatogram, on position corresponding with reference medicine chromatography, phase is shown
With the spot of color.
(4) method that radix cyathulae is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking 18 ball this product, adds ethyl alcohol 40ml, at ultrasound
Reason 30 minutes, filtration, filtrate is evaporated, and residue adds diethyl ether 20ml, and shaking is extracted, volatilized, and residue adds ethyl acetate 1ml to make to dissolve,
As test solution；Radix cyathulae control medicinal material 2.5g separately is taken, adds ethyl alcohol 30ml, comparison medicine is made in sample solution preparation method
Product solution.According to TLC test, 7 μ l of test solution, 4 μ l of control medicinal material solution are drawn, is put respectively thin in same silica G
On laminate, with toluene: ethyl acetate: formic acid (18: 13: 2) is solvent, is unfolded, and takes out, dries, and is sprayed with 1% tri-chlorination
Iron: it is clear that the mixed solution of 1% potassium ferricyanide (1: 1) is heated to spot development.In sample chromatogram, with control medicinal material
On the corresponding position of chromatography, the spot of same color is shown.
Cyasterone reference substance is taken, adds methanol that solution of every 1ml containing 0.5mg is made, as reference substance solution.According to thin layer color
Spectrometry test, draws 8 μ l of test solution and 5 μ l of reference substance solution, is put respectively on same silica gel g thin-layer plate, with chloroform-the third
Ketone-formic acid (15: 4: 1) is solvent, is unfolded, and takes out, dries, and sprays with 10% ethanol solution of sulfuric acid, is heated to spot at 105 DEG C
Colour developing is clear, sets and inspects under ultraviolet lamp.In sample chromatogram, on position corresponding with reference medicine chromatography, identical face is shown
The spot of color.
Assay:
Chromatographic condition and system suitability:
Flow stationary phase: selecting octadecylsilane chemically bonded silica is the chromatographic column of filler；
Mobile phase: the acetonitrile of volume ratio 45: 55: methanol (5: 1) -0.02mol/l potassium dihydrogen phosphate mixing
Solvent adjusts pH value to 4.0 as mobile phase (triethylamine that 3ml is added in every 1000ml), with phosphoric acid solution；
Detection wavelength: 300nm；
Column temperature: 30 DEG C；
Flow velocity: 1.0ml/min.
The preparation of test solution: taking this product for being equivalent to raw medicinal herbs 17.56g, accurately weighed, finely ground, takes about 1.8g,
It is accurately weighed, it sets in stuffed conical flask, 50% methanol 25ml is added in precision, and weighed weight ultrasonic extraction 40 minutes, is let cool, then
Weighed weight supplies the weight of less loss with 50% methanol, filtration, discards primary filtrate, and remaining filtrate is 0.45 μm micro- by aperture
Hole filter membrane filters to get test solution.
The preparation of reference substance solution: precision weighs Imperatorin reference substance, accurately weighed, adds methanol that every 1ml is made and contains
The solution of 20 μ g, shake up to get.
Measuring method: accurate absorption reference substance solution and each 10 μ l of test solution respectively, injection liquid chromatograph, measurement,
To obtain the final product.
The Chinese medicine preparation for being equivalent to 1g raw medicinal herbs contains the root of Dahurain angelica with Imperatorin (C18H14O4Meter) meter, it must not be less than
0.067mg。
Claims (2)
1. a kind of detection method for the alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding, which is characterized in that the detection method includes
The discrimination method and content assaying method of alizarin root of Dahurian angelica Chinese materia medica preparation；Identification be in alizarin root of Dahurian angelica Chinese materia medica preparation madder, rubimaillin,
Purpurin, Radix Notoginseng, notoginsenoside, ginsenoside, the root of Dahurain angelica, Imperatorin, Isomperatorin, radix cyathulae, cyasterone
Indentification by TLC；The discrimination method are as follows:
Character: the alizarin root of Dahurian angelica Chinese materia medica preparation of the treatment uterus bleeding is brown, gas fragrance, acrid flavour, slight bitter；
Identify:
(1) method that madder is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking in the alizarin root of Dahurian angelica for being equivalent to 15.25~20.55g of raw medicinal herbs
Medicine preparation removes coating, finely ground, and 20~40ml of ether is added, and places 1 hour, and constantly shaking is extracted, and filtration, filtrate is flung to, residual
Slag adds ethyl acetate 1ml to make to dissolve, spare as madder test solution；Madder control medicinal material 0.8~1.5g of powder separately is taken, together
Control medicinal material solution is made in madder sample solution preparation method；It is tested according to thin-layered chromatography, draws each 5 μ of above two solution
L puts the cyclohexane-acetone-two for being 8~15: 4~10: 1~3: 4~10 with volume ratio on same silica gel g thin-layer plate respectively
Ethamine-water be solvent expansion, take out, dry, set and inspected under 365nm ultraviolet lamp, in sample chromatogram, with comparison medicine wood color
It composes on corresponding position, shows the spot of same color；
Taking rubimaillin is reference substance, and the solution that 1ml 0.1mg containing rubimaillin is made in methanol is added, molten as reference substance
Liquid；It is tested according to thin-layered chromatography, draws reference substance solution and each 5 μ l of the madder test solution, put respectively in same silica G
It is 20~25: 3~6: 0.1~0.8 petroleum ether-ethyl acetate-glacial acetic acid as solvent exhibition using volume ratio on lamellae
It opens, takes out, dry, spray is with dinitrophenylhydrazine ethyl alcohol test solution, in sample chromatogram, at the position corresponding to the chromatogram of the reference substance,
The spot of aobvious same color；The petroleum ether that the step uses is 60~90 DEG C；
Taking purpurin is reference substance, methanol is added, every 40 μ g of 1ml hydroxyl rubican is made, as reference substance solution；According to
Thin-layered chromatography test, draws reference substance solution and each 5 μ l of the madder test solution, is put respectively in same silica G thin layer
It is 15~20: 5~12: 1~2 ethyl acetate-acetones-formic acid as solvent using volume ratio on plate, is unfolded, takes out, dry, set
It is inspected under 365nm ultraviolet lamp, in sample chromatogram, at the position corresponding to the chromatogram of the reference substance, shows the spot of same color；
(2) method that Radix Notoginseng is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking the alizarin root of Dahurian angelica Chinese medicine for being equivalent to 13.6~18.6g of raw medicinal herbs
Preparation removes coating, finely ground, and 15~30ml of methanol solution of 5% formic acid is added, is heated to reflux 30 minutes, lets cool, filters, filtrate
It flings to, residue adds methanol 2ml to make to dissolve, spare as Radix Notoginseng test solution；Separately take Radix Notoginseng control medicinal material powder 0.8~
Control medicinal material solution is made with Radix Notoginseng sample solution preparation method in 1.5g；It tests, draws above two molten according to thin-layered chromatography
Each 5 μ l of liquid puts respectively on same silica gel g thin-layer plate, is for 8~12: 6~9: 1~3 benzene-acetic ether-methanoic acid with volume ratio
Solvent is unfolded, and takes out, dries, set and inspect under 365nm ultraviolet lamp, in sample chromatogram, corresponding with reference medicine chromatography
On position, the spot of same color is shown；
Take Panax Notoginseng saponin R1With Panax Notoginseng saponin R g1Reference substance adds methanol that solution of every 1ml containing 1mg is made, as reference substance solution；
It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of Radix Notoginseng test solution, put respectively in same silica gel g thin-layer plate
On, using the upper solution that volume ratio is 5~12: 3~7: 0.5~1.5 chloroform-methanols-glacial acetic acid as solvent, it is unfolded, takes out,
It dries, sprays with ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C；In sample chromatogram, with reference substance chromatography pair
On the position answered, the spot of same color is shown；
Ginsenoside Rb1's reference substance and ginsenoside Re's reference substance are taken, adds methanol that mixed solution of every 1ml containing 0.5mg is made, is made
For reference substance solution；It is tested according to thin-layered chromatography, draws reference substance solution and each 3 μ l of Radix Notoginseng test solution is put in same respectively
On silica gel g thin-layer plate, with volume ratio for 10~18: 35~45: 17~25: 8~12 chloroforms-acetate-methanol-water 10
DEG C lower layer's solution arranged below is solvent, is unfolded, and takes out, dries, and it is aobvious to be heated to spot at 105 DEG C with sulfuric acid solution for spray
Color is clear；In sample chromatogram, at the position corresponding to the chromatogram of the reference substance, identical colored speckles are shown；
(3) method that the root of Dahurain angelica is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking the alizarin root of Dahurian angelica Chinese medicine for being equivalent to 14.6~19.8g of raw medicinal herbs
Preparation removes coating, finely ground, adds 10~40ml of petroleum ether, and shaking is extracted 30 minutes, and filtration, filter residue is spare, and filtrate flings to petroleum
Ether is spare as root of Dahurain angelica test solution to 2ml；The root of Dahurain angelica 0.4~0.9g of control medicinal material separately is taken, adds petroleum ether (60~90 DEG C) 5
~10ml, shaking are extracted 30 minutes, are stood, and supernatant is tested as control medicinal material solution according to thin-layered chromatography, are drawn above-mentioned
Each 5 μ l of two kinds of solution is put respectively on same silica gel g thin-layer plate, with volume ratio for 12~20: 2~5: 0.1~0.8 petroleum ether-
Ethyl acetate-glacial acetic acid is solvent, is unfolded at 25 DEG C or less, takes out, dries, set and inspect under 365nm ultraviolet lamp；Test sample
In chromatography, on position corresponding with reference medicine chromatography, the spot of same color is shown；The petroleum ether that the step uses is 60
~90 DEG C；
Imperatorin reference substance and Isomperatorin reference substance are taken, it is molten that the mixing that every 1ml contains 1mg is made in addition ethyl acetate
Liquid, as reference substance solution；It is tested according to thin-layered chromatography, draws reference substance solution and each 4 μ l of root of Dahurain angelica test solution, respectively point
It is 5~9: 2~5: 0~1 petroleum ether-ether-formic acid as solvent using volume ratio, at 25 DEG C on same silica gel g thin-layer plate
It is unfolded below, takes out, dry, set and inspected under ultraviolet lamp；In sample chromatogram, at the position corresponding to the chromatogram of the reference substance,
The spot of aobvious same color；The petroleum ether that the step uses is 30~60 DEG C；
(4) method that radix cyathulae is identified in alizarin root of Dahurian angelica Chinese materia medica preparation prescription is: taking in the alizarin root of Dahurian angelica for being equivalent to 12.7~18.9g of raw medicinal herbs
Medicine preparation, adds 30~50ml of ethyl alcohol, is ultrasonically treated 30 minutes, and filtration, filtrate is evaporated, and residue adds diethyl ether 15~25ml, and shaking mentions
It takes, volatilizes, residue adds ethyl acetate 1ml to make to dissolve, as radix cyathulae test solution；Separately take radix cyathulae control medicinal material 1~
4g adds 20~40ml of ethyl alcohol, and control medicinal material solution is made with radix cyathulae sample solution preparation method；According to TLC test,
6~8 μ l of radix cyathulae test solution, 3~5 μ l of control medicinal material solution are drawn, is put respectively on same silica G lamellae, with body
Product is than being 10~25: 8~18: 1~3 toluene: ethyl acetate: formic acid is solvent, is unfolded, and takes out, dries, and is sprayed with volume ratio
For 1: 1% ferric trichloride: it is clear that the mixed solution of 1% potassium ferricyanide is heated to spot development；In sample chromatogram, with compare
On the corresponding position of medicinal material chromatography, the spot of same color is shown；
Cyasterone reference substance is taken, adds methanol that solution of every 1ml containing 0.5mg is made, as reference substance solution；According to thin-layered chromatography
5~10 μ l of radix cyathulae test solution and 5 μ l of reference substance solution is drawn in test, is put respectively on same silica gel g thin-layer plate, with
Volume ratio is that 13~18: 3~5: 1~2 chloroforms-acetone-formic acid is solvent, is unfolded, and takes out, dries, and is sprayed with 10% sulfuric acid ethyl alcohol
Solution, it is clear to be heated to spot development at 105 DEG C, sets and inspects under ultraviolet lamp；In sample chromatogram, with reference substance chromatography phase
On the position answered, the spot of same color is shown；
The content assaying method of the madder root of Dahurian angelica Chinese materia medica preparation, including using white in high effective liquid chromatography for measuring madder root of Dahurian angelica Chinese materia medica preparation
The amount of Imperatorin contained by the root of Dahurian angelica；
Chromatographic condition and system suitability:
Flow stationary phase: selecting octadecylsilane chemically bonded silica is the chromatographic column of filler；
Mobile phase: using the mixed solvent that component A and component B that volume ratio is (30: 70)~(65: 35) are constituted as stream
Dynamic phase；The component A is the acetonitrile and methanol solution that volume ratio is 5: 1, and component B is 0.01~0.05mol/L phosphoric acid
Dihydro potassium solution or sodium dihydrogen phosphate or ammonium dihydrogen phosphate, every 1000ml flowing are added to the triethylamine of 3ml, use phosphorus
Acid solution adjusts pH value to 3.0~4.5；
Detection wavelength: 275~315nm；
Column temperature: 25~45 DEG C；
Flow velocity: 0.8~1.2ml/min of flow velocity；
The preparation of test solution: taking the alizarin root of Dahurian angelica Chinese materia medica preparation for being equivalent to raw medicinal herbs 17.56g, accurately weighed, finely ground, takes
1.8g, it is accurately weighed, it sets in stuffed conical flask, 50% methanol 50ml, weighed weight, ultrasonic extraction 30~40 is added in precision
Minute, it lets cool, then weighed weight, the weight of less loss is supplied with 50% methanol, filter, discard primary filtrate, remaining filtrate through hole
Diameter is 0.45 μm of miillpore filter filtration to get test solution；
The preparation of reference substance solution: precision weighs Imperatorin reference substance, accurately weighed, adds methanol that every 1ml is made and contains 20 μ g
Solution, shake up to get；
Measuring method: it is accurate respectively to draw reference substance solution and each 10 μ l of test solution, inject liquid chromatograph, measurement to get.
2. a kind of detection method for the alizarin root of Dahurian angelica Chinese materia medica preparation for treating uterus bleeding, feature shown according to claim 1
Be: the chromatographic condition and component A in system suitability and component B using volume ratio be 45: 55 mix after as flowing
The triethylamine of 3ml is added in phase and every 1000ml, adjusts pH value to 4.0 with phosphoric acid solution；The component A is that volume ratio is
5: 1 acetonitrile and methanol solution, the component B are 0.02mol/l potassium dihydrogen phosphates；The Detection wavelength 300nm；
The column temperature is 30 DEG C, and the flow velocity is 1.0ml/min.
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