CN1670525A - Method for measuring fingerprint pattern of compound salvia dropping pills - Google Patents
Method for measuring fingerprint pattern of compound salvia dropping pills Download PDFInfo
- Publication number
- CN1670525A CN1670525A CN 200510055264 CN200510055264A CN1670525A CN 1670525 A CN1670525 A CN 1670525A CN 200510055264 CN200510055264 CN 200510055264 CN 200510055264 A CN200510055264 A CN 200510055264A CN 1670525 A CN1670525 A CN 1670525A
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- acetic acid
- glacial acetic
- rsd
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 104
- 239000006187 pill Substances 0.000 title claims abstract description 104
- 235000017276 Salvia Nutrition 0.000 title claims abstract description 27
- 240000007164 Salvia officinalis Species 0.000 title claims description 50
- 238000000034 method Methods 0.000 title abstract description 33
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 167
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 124
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 96
- 229960000583 acetic acid Drugs 0.000 claims abstract description 83
- 239000012362 glacial acetic acid Substances 0.000 claims abstract description 82
- 239000000243 solution Substances 0.000 claims abstract description 72
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims abstract description 46
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 39
- 238000012360 testing method Methods 0.000 claims abstract description 29
- 239000000706 filtrate Substances 0.000 claims abstract description 25
- 239000000945 filler Substances 0.000 claims abstract description 21
- 239000007788 liquid Substances 0.000 claims abstract description 21
- 229910021529 ammonia Inorganic materials 0.000 claims abstract description 20
- 239000012528 membrane Substances 0.000 claims abstract description 17
- 238000010828 elution Methods 0.000 claims abstract description 9
- 239000009524 danshen dripping Substances 0.000 claims description 73
- 239000007864 aqueous solution Substances 0.000 claims description 40
- 238000002360 preparation method Methods 0.000 claims description 35
- 238000005303 weighing Methods 0.000 claims description 23
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 21
- 238000003556 assay Methods 0.000 claims description 20
- 238000005406 washing Methods 0.000 claims description 18
- 238000011010 flushing procedure Methods 0.000 claims description 16
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 15
- 238000004090 dissolution Methods 0.000 claims description 15
- 239000012530 fluid Substances 0.000 claims description 12
- 238000004811 liquid chromatography Methods 0.000 claims description 12
- 239000006210 lotion Substances 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 7
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 7
- 239000000377 silicon dioxide Substances 0.000 claims description 7
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 6
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 6
- 235000015096 spirit Nutrition 0.000 claims description 5
- 239000000203 mixture Substances 0.000 abstract description 41
- 239000000126 substance Substances 0.000 abstract description 40
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 13
- 238000003908 quality control method Methods 0.000 abstract description 11
- 238000001514 detection method Methods 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 2
- 238000002347 injection Methods 0.000 abstract description 2
- 239000007924 injection Substances 0.000 abstract description 2
- 241001072909 Salvia Species 0.000 abstract 7
- 239000003480 eluent Substances 0.000 abstract 1
- 230000014759 maintenance of location Effects 0.000 description 82
- 244000131316 Panax pseudoginseng Species 0.000 description 64
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 64
- 239000012071 phase Substances 0.000 description 63
- 239000003814 drug Substances 0.000 description 37
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 36
- 235000005412 red sage Nutrition 0.000 description 30
- 238000010521 absorption reaction Methods 0.000 description 25
- 241000180649 Panax notoginseng Species 0.000 description 18
- 235000003143 Panax notoginseng Nutrition 0.000 description 17
- 239000000463 material Substances 0.000 description 15
- 244000132619 red sage Species 0.000 description 15
- 235000011135 Salvia miltiorrhiza Nutrition 0.000 description 14
- 239000000284 extract Substances 0.000 description 14
- 230000000694 effects Effects 0.000 description 13
- 229930182494 ginsenoside Natural products 0.000 description 12
- 229940089161 ginsenoside Drugs 0.000 description 12
- 239000000470 constituent Substances 0.000 description 10
- AIGAZQPHXLWMOJ-UHFFFAOYSA-N tanshinone IIA Natural products C1=CC2=C(C)C=CC=C2C(C(=O)C2=O)=C1C1=C2C(C)=CO1 AIGAZQPHXLWMOJ-UHFFFAOYSA-N 0.000 description 10
- 229940079593 drug Drugs 0.000 description 9
- 239000007791 liquid phase Substances 0.000 description 9
- 239000012153 distilled water Substances 0.000 description 7
- 239000000047 product Substances 0.000 description 6
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 6
- 229930182490 saponin Natural products 0.000 description 6
- 150000007949 saponins Chemical class 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- DTGKSKDOIYIVQL-WEDXCCLWSA-N (+)-borneol Chemical compound C1C[C@@]2(C)[C@@H](O)C[C@@H]1C2(C)C DTGKSKDOIYIVQL-WEDXCCLWSA-N 0.000 description 5
- REPVLJRCJUVQFA-UHFFFAOYSA-N (-)-isopinocampheol Natural products C1C(O)C(C)C2C(C)(C)C1C2 REPVLJRCJUVQFA-UHFFFAOYSA-N 0.000 description 5
- 229940116229 borneol Drugs 0.000 description 5
- CKDOCTFBFTVPSN-UHFFFAOYSA-N borneol Natural products C1CC2(C)C(C)CC1C2(C)C CKDOCTFBFTVPSN-UHFFFAOYSA-N 0.000 description 5
- DTGKSKDOIYIVQL-UHFFFAOYSA-N dl-isoborneol Natural products C1CC2(C)C(O)CC1C2(C)C DTGKSKDOIYIVQL-UHFFFAOYSA-N 0.000 description 5
- 208000024172 Cardiovascular disease Diseases 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- HYXITZLLTYIPOF-UHFFFAOYSA-N Tanshinone II Natural products O=C1C(=O)C2=C3CCCC(C)(C)C3=CC=C2C2=C1C(C)=CO2 HYXITZLLTYIPOF-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 208000026106 cerebrovascular disease Diseases 0.000 description 4
- 230000002526 effect on cardiovascular system Effects 0.000 description 4
- 241000411851 herbal medicine Species 0.000 description 4
- AZEZEAABTDXEHR-UHFFFAOYSA-M sodium;1,6,6-trimethyl-10,11-dioxo-8,9-dihydro-7h-naphtho[1,2-g][1]benzofuran-2-sulfonate Chemical compound [Na+].C12=CC=C(C(CCC3)(C)C)C3=C2C(=O)C(=O)C2=C1OC(S([O-])(=O)=O)=C2C AZEZEAABTDXEHR-UHFFFAOYSA-M 0.000 description 4
- PAFLSMZLRSPALU-MRVPVSSYSA-N (2R)-3-(3,4-dihydroxyphenyl)lactic acid Chemical compound OC(=O)[C@H](O)CC1=CC=C(O)C(O)=C1 PAFLSMZLRSPALU-MRVPVSSYSA-N 0.000 description 3
- PAFLSMZLRSPALU-QMMMGPOBSA-N Danshensu Natural products OC(=O)[C@@H](O)CC1=CC=C(O)C(O)=C1 PAFLSMZLRSPALU-QMMMGPOBSA-N 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 3
- PAFLSMZLRSPALU-UHFFFAOYSA-N Salvianic acid A Natural products OC(=O)C(O)CC1=CC=C(O)C(O)=C1 PAFLSMZLRSPALU-UHFFFAOYSA-N 0.000 description 3
- 238000009835 boiling Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 3
- 239000008108 microcrystalline cellulose Substances 0.000 description 3
- 229940016286 microcrystalline cellulose Drugs 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- IBGBGRVKPALMCQ-UHFFFAOYSA-N 3,4-dihydroxybenzaldehyde Chemical compound OC1=CC=C(C=O)C=C1O IBGBGRVKPALMCQ-UHFFFAOYSA-N 0.000 description 2
- YQUVCSBJEUQKSH-UHFFFAOYSA-N 3,4-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 description 2
- XYKZSUXWBGUGQV-UHFFFAOYSA-N 4,8-dimethylnaphtho[2,1-f][1]benzofuran-7,11-dione Chemical compound CC1=CC=CC2=C(C(C=3OC=C(C=3C3=O)C)=O)C3=CC=C21 XYKZSUXWBGUGQV-UHFFFAOYSA-N 0.000 description 2
- 241000208340 Araliaceae Species 0.000 description 2
- 206010013786 Dry skin Diseases 0.000 description 2
- 235000008100 Ginkgo biloba Nutrition 0.000 description 2
- 244000194101 Ginkgo biloba Species 0.000 description 2
- 239000009636 Huang Qi Substances 0.000 description 2
- 229930183118 Tanshinone Natural products 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- QMNWISYXSJWHRY-YLNUDOOFSA-N astragaloside IV Chemical compound O1[C@H](C(C)(O)C)CC[C@]1(C)[C@@H]1[C@@]2(C)CC[C@]34C[C@]4(CC[C@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)CO4)O)C4(C)C)[C@H]4[C@@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)C[C@H]3[C@]2(C)C[C@@H]1O QMNWISYXSJWHRY-YLNUDOOFSA-N 0.000 description 2
- QMNWISYXSJWHRY-BCBPIKMJSA-N astragaloside IV Natural products CC(C)(O)[C@@H]1CC[C@@](C)(O1)[C@H]2[C@@H](O)C[C@@]3(C)[C@@H]4C[C@H](O[C@@H]5O[C@H](CO)[C@H](O)[C@@H](O)[C@H]5O)[C@H]6C(C)(C)[C@H](CC[C@@]67C[C@@]47CC[C@]23C)O[C@@H]8OC[C@@H](O)[C@H](O)[C@H]8O QMNWISYXSJWHRY-BCBPIKMJSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 208000029078 coronary artery disease Diseases 0.000 description 2
- PFKIBRPYVNVMRU-UHFFFAOYSA-N cyclosieversioside F Natural products CC(C)(O)C1COC(C)(C1)C2C(O)CC3(C)C4CC(OC5OC(CO)C(O)C(O)C5O)C6C(C)(C)C(CCC67CC47CCC23C)OC8OCC(O)C(O)C8O PFKIBRPYVNVMRU-UHFFFAOYSA-N 0.000 description 2
- OORMXZNMRWBSTK-LGFJJATJSA-N dammarane Chemical compound C1CCC(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@H]([C@H](C)CCCC(C)C)[C@H]4CC[C@@H]3[C@]21C OORMXZNMRWBSTK-LGFJJATJSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000004744 fabric Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000008899 fufang danshen Substances 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 239000011122 softwood Substances 0.000 description 2
- 235000020985 whole grains Nutrition 0.000 description 2
- LJEROUHPDCFEGJ-VVTLTYDVSA-N (1r,3r,4s)-4,7,9-trihydroxy-1,3-dimethyl-3,4-dihydro-1h-benzo[g]isochromene-5,10-dione Chemical compound O=C1C2=CC(O)=CC(O)=C2C(=O)C2=C1[C@H](O)[C@@H](C)O[C@@H]2C LJEROUHPDCFEGJ-VVTLTYDVSA-N 0.000 description 1
- XDUXBBDRILEIEZ-LJQANCHMSA-N (6s)-6-(hydroxymethyl)-1,6-dimethyl-8,9-dihydro-7h-naphtho[1,2-g][1]benzofuran-10,11-dione Chemical compound O=C1C(=O)C2=C3CCC[C@](C)(CO)C3=CC=C2C2=C1C(C)=CO2 XDUXBBDRILEIEZ-LJQANCHMSA-N 0.000 description 1
- WTPPRJKFRFIQKT-UHFFFAOYSA-N 1,6-dimethyl-8,9-dihydronaphtho[1,2-g][1]benzofuran-10,11-dione;1-methyl-6-methylidene-8,9-dihydro-7h-naphtho[1,2-g][1]benzofuran-10,11-dione Chemical compound O=C1C(=O)C2=C3CCCC(=C)C3=CC=C2C2=C1C(C)=CO2.O=C1C(=O)C2=C3CCC=C(C)C3=CC=C2C2=C1C(C)=CO2 WTPPRJKFRFIQKT-UHFFFAOYSA-N 0.000 description 1
- MTJGVAJYTOXFJH-UHFFFAOYSA-N 3-aminonaphthalene-1,5-disulfonic acid Chemical compound C1=CC=C(S(O)(=O)=O)C2=CC(N)=CC(S(O)(=O)=O)=C21 MTJGVAJYTOXFJH-UHFFFAOYSA-N 0.000 description 1
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 244000247747 Coptis groenlandica Species 0.000 description 1
- 235000002991 Coptis groenlandica Nutrition 0.000 description 1
- GVKKJJOMQCNPGB-JTQLQIEISA-N Cryptotanshinone Chemical compound O=C1C(=O)C2=C3CCCC(C)(C)C3=CC=C2C2=C1[C@@H](C)CO2 GVKKJJOMQCNPGB-JTQLQIEISA-N 0.000 description 1
- GVKKJJOMQCNPGB-UHFFFAOYSA-N Cryptotanshinone Natural products O=C1C(=O)C2=C3CCCC(C)(C)C3=CC=C2C2=C1C(C)CO2 GVKKJJOMQCNPGB-UHFFFAOYSA-N 0.000 description 1
- 244000301850 Cupressus sempervirens Species 0.000 description 1
- 206010013709 Drug ineffective Diseases 0.000 description 1
- SHWNNYZBHZIQQV-UHFFFAOYSA-J EDTA monocalcium diisodium salt Chemical compound [Na+].[Na+].[Ca+2].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O SHWNNYZBHZIQQV-UHFFFAOYSA-J 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 240000007711 Peperomia pellucida Species 0.000 description 1
- 235000012364 Peperomia pellucida Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- XDUXBBDRILEIEZ-UHFFFAOYSA-N Tanshinone IIB Natural products O=C1C(=O)C2=C3CCCC(C)(CO)C3=CC=C2C2=C1C(C)=CO2 XDUXBBDRILEIEZ-UHFFFAOYSA-N 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 208000029162 bladder disease Diseases 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 150000004141 diterpene derivatives Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 208000031169 hemorrhagic disease Diseases 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 238000004452 microanalysis Methods 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 208000031225 myocardial ischemia Diseases 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 244000138993 panchioli Species 0.000 description 1
- -1 phenol aldehyde Chemical class 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 238000003822 preparative gas chromatography Methods 0.000 description 1
- 229940126532 prescription medicine Drugs 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 229960003371 protocatechualdehyde Drugs 0.000 description 1
- 238000001303 quality assessment method Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 229920002545 silicone oil Polymers 0.000 description 1
- KZJWDPNRJALLNS-VJSFXXLFSA-N sitosterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CC[C@@H](CC)C(C)C)[C@@]1(C)CC2 KZJWDPNRJALLNS-VJSFXXLFSA-N 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000008736 traumatic injury Effects 0.000 description 1
- 229930182493 triterpene saponin Natural products 0.000 description 1
- 208000026533 urinary bladder disease Diseases 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a method for measuring fingerprint pattern of compound salvia dropping pills, a method for establishing HPLC fingerprint pattern of compound salvia dropping pills by experiments and a standard fingerprint pattern thereof. A testing solution of salvia dropping pills is prepared firstly: 1.0g salvia dropping pills are weighed and dissolved in 10mL 4% ammonia with ultrasonic and then filtered by 0.45 mu m filter membrane. 5mL filtrate is taken and passed through a C-18 column with 500mg filler, rinsed by 20mL solution containing 20% methanol and 4% ammonia with the rinsing liquid discarded, then rinsed by 20mL water with the water rinsing liquid discarded and then eluted by methanol. The methanol eluent is collected in a volumetric flask of 5ml, volume metered and centrifuged for use with injection volume to be 20 mu L. Gradient elution is adopted with mobile phase A to be 0.01% glacial acetic acid solution and mobile phase B to be acetonitrile solution containing 0.01% glacial acetic acid; flow rate is 0.8ml.min[-1], detection wavelength is 203nm, column temperature is 30DEG C; the testing solution is determined by high performance liquid chromatography method to obtain the fingerprint pattern of compound salvia dropping pills. The method can obtain fingerprint pattern with high repeatability and be used as method for determining chemical compositions in salvia dropping pills and quality control method for identifying compound salvia dropping pills.
Description
Technical field
The present invention relates to a kind of assay method of fingerprint pattern of compound salvia dropping pills, specifically use high performance liquid chromatography (HPLC) fingerprint spectrum method to measure a kind of method of effective constituent in the compound danshen dripping pills.
Background technology
Cardiovascular and cerebrovascular disease is the serious harm mankind's a common disease.In recent years, since the development of society, the variation of work, life, dietary structure and environment etc., and cardiovascular and cerebrovascular disease is in rising trend.Treatment for angiocardiopathy, though the action intensity of the single target spot of Chinese medicine is lower than Western medicine, but its multipath, many target spots, dynamically wholistic therapy, characteristic that toxic and side effect is little then are far from Western medicine and can reach, and the combined therapy effect of the Chinese patent drug of determined curative effect will surpass Western medicine.Compound red sage root preparation is used for treating cardiovascular and cerebrovascular disease more, for example Fufang Danshen Pian, compound danshen dripping pills, coronary heart disease drop pill etc.These compound red sage root preparations (all containing the red sage root, pseudo-ginseng) are different because of its prescription, and perhaps the formula rate difference is perhaps extracted the process for purification difference, and perhaps formulation difference, result of treatment be difference to some extent also.In addition, because the method for quality control of these compound red sage root preparations is comprehensive inadequately, be difficult to characterize their physicochemical characteristic comprehensively.Therefore, extraction process for purification, the method for quality control to compound red sage root preparation is modified into the problem into people's active research.
The red sage root is the dry root and rhizome of labiate red sage root Salvia miltiorrhiza Bge..All produce in most of area, the whole nation.Because the red sage root causes quality uneven because of kind, the place of production, picking time are different, thereby its manufactured goods quality of stability also is difficult to guarantee.At present, to the evaluation of the red sage root and compound preparation thereof, choose one, two active component or the index components of the red sage root often and carry out assay, and how much judge quality with its content.For example, with tanshinone IIA content (Chinese Pharmacopoeia 2000 version one one 58 pages) or content of Danshensu (Zhang Youqin etc., traditional Chinese medicine journal, 2000,28 (3): 68) wait the quality of differentiating red rooted salvia; Judge red sage root kind and place of production situation (Qiu Feijun, contemporary Chinese application pharmacy, 1998,15 (5): 16 with tanshinone; Hu Shilin etc., CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1999,24 (12): 721); With content of Danshensu (Yan Changkai etc., Chinese Hospitals pharmaceutical journal, 2000,600), protocatechualdehyde content (Zheng end crystalline substance etc., Chinese Pharmaceutical Affairs, 2,000 20 (10):, 254) or tanshinone IIA content (Lin Weizhong etc. 14 (4):, Chinese patent drug, 766) etc. 2000,22 (11): the quality of differentiating compound red sage root preparation, differentiate the quality (Shao Shuijuan of the Fufang Danshen Pian that different manufacturers is produced with tanshinone IIA content, China's medicine company, 2000,9 (7): 27) etc.The chemical constitution of the known red sage root has tens kinds, and its liposoluble constituent mostly is quinoid reddish yellow material greatly, as Tanshinone I, IIA, IIB, and red sage root quinone A, B, C, red sage root acid potassium fat, isotanshinone I, II, Cryptotanshinone etc.Its water soluble ingredient mostly is phenol aldehyde, phenol acid, diterpenoid acid greatly, as succinic acid, salviandic acid A, B, C, and 3,4-dihydroxy-benzoic acid etc.In addition, also isolate cupreol, vitamin E etc. (Wang Baixiang chief editor, traditional Chinese medical science liver and bladder disease is learned, first published, Chinese Medicine science and technology publishing house,, 96 pages in 1993).Pseudo-ginseng is Araliaceae (Araliaceae) plant pseudo-ginseng Panaxnotoginseng (Burk.) F.H.Chen, dry root, main product in Yunnan, ground such as Guangxi and Sichuan, be the special product medicinal material of China's preciousness, conventional Chinese medicine.Its flavor is sweet, and little hardship is warm in nature, returns liver, kidney channel, has the effect of diffusing stasis of blood hemostasis, swelling and pain relieving, and tradition is used for the treatment of traumatic injury and various hemorrhagic disease.Studies show that pseudo-ginseng mainly contains chemical constitutions such as saponin, polysaccharide, amino acid, wherein saponin partly is the material base that pseudo-ginseng blood-circulation-invigovating stagnation resolvation effect is used, and is the main effective constituent of pseudo-ginseng.Notoginseng total saponin contains panaxoside R
B1, R
B2, R
c, R
d, R
e, R
f, R
G1, R
G2, B
H1, arasaponin R
1, R
2, R
3, R
4, R
6Deng kind of saponin component surplus 20.These compositions all belong to dammarane type [Dammarane type] tetracyclic triterpene saponin, wherein panaxoside R
B1, R
G1, arasaponin R
1Be 3 the highest compositions of content, arasaponin R
1It is the pseudo-ginseng compound of representative feature.
That traditional Chinese medicine fingerprint is meant is common in certain Chinese crude drug or the Chinese patent drug, have distinctive certain class or the chromatogram of number constituents or the collection of illustrative plates of spectrum.Do not have under the clear and definite situation in the present stage Effective Components of Chinese Herb overwhelming majority, traditional Chinese medicine fingerprint has great importance for the quality of effective control Chinese crude drug or Chinese patent drug.The Japan main manufacturing enterprise of Chinese prescription medicine just adopts the high-efficiency liquid-phase fingerprint control of quality in enterprises in the eighties in 20th century.Germany, France find that the medical function of ginkgo biloba p.e is extract gained material group's mass action result in the process that ginkgo biloba p.e is developed jointly, and to the quality control of such integral body, also adopt the high-efficiency liquid-phase fingerprint method.In the plant herbal medicine guide of formulating U.S. FDA recent years clearly the method for quality control (FDA.Guidance of Industry:Botanical Drug (Draft) .2000 August) of finger-print as the compounding substances group.Along with going deep into of research, it is found that, as the product of putting into practice of theory of traditional Chinese medical science, Chinese medicine, especially herbal mixture, wherein contained arbitrary composition all can not be represented its whole curative effect.People recognize that gradually the existing quality standard with reference to Western medicine (synthetic drug) quality control pattern can not reflect the quality of Chinese medicine inherence rightly.From development trend, from existing quality control pattern to a kind of comprehensive, macroscopic view, quantifiable discriminating combines with main active constituent content measuring is the trend that develops.
The act.std of medicine quality evaluated is to utilize spectrum or the discriminating of chromatogram means and measure a certain or several effective constituents, active component or index components, and the routine inspection project of pharmacopeia regulation.Record 602 kinds of medicinal materials and patent medicine kind altogether as Chinese Pharmacopoeia 2000 version [an one].Wherein have 992 thin-layer chromatographys to differentiate that 308 kinds have assay (volumetric method, spectroscopic methodology, liquid phase chromatography, vapor-phase chromatography and thin layer chromatography (TLC) scanning), most of kinds have general inspection item.Obviously, the setting of these quality standards is the patterns of having imitated chemicals.The German herbal medicine monograph that other country edits as the Chinese medicine in Britain, India, U.S.'s herbal medicine allusion quotation, the Pharmacopeia of Japan and German Commission E etc. has also adopted essentially identical content.For chemicals, its effective component is the simplification compound of clear in structure, and structure-activity relationship is clear and definite, and its content and purity are directly expressed it and effectively reached security.Yet the characteristics of middle medical drugs are compound compatibilities, any single effectively or the content of active component height all can not express its whole curative effect.For example, the contained Astragaloside IV (aastraga losideIV) of the Radix Astragali is the current discriminating of quality standard and the most common target of assay of being selected as, but not according to clearly getting in touch that the function that proves the Astragaloside IV and the Radix Astragali cures mainly.Equally, the coptis, golden cypress, radix berberidis all contain girder alkali, and be general all with its target as detection, completely different but three's function cures mainly.The situation of compound preparation is just complicated more.The traditional Chinese medical science is this not to be the comprehensive quality assessment means that man-to-man nonlinear theory and practice explanation traditional Chinese medicine quality should adopt certain macroscopic view.
Compound danshen dripping pills is to be the dropping pill formulation that primary raw material is made by the red sage root, pseudo-ginseng, be used for the treatment of cardiovascular and cerebrovascular disease, coronary heart disease, angina pectoris, myocardial ischemia, all kinds of diseases that microcirculation disorder caused etc. clinically, its result of treatment has obtained clinical checking, and whether can guarantee content of effective in the quality of medicine and the compound danshen dripping pills, be the basis of decision compound danshen dripping pills curative effect.If with one, the active component of two kind of red sage root illustrates the inherent quality of compound danshen dripping pills, has certain one-sidedness, said nothing of the index components of no drug effect.Control the effect of compound danshen dripping pills, only at one, two chemical constitutions characterize and control is not enough, must be controlled its material group integral body.So, except " micro-analysis ", also should characterize traditional Chinese medicine quality on the whole effectively with certain " macroanalysis " method.Finger-print is become a consensus of the international community at present as Chinese herbal medicine and extraction of substance amount control method thereof.Now, more to the assay method of active component such as tanshinone, danshensu etc. in the red sage root, to active component in the pseudo-ginseng such as arasaponin R
G1, panaxoside R
G1More Deng assay method, but how can the macroscopic quality control method that the composite salvia dropping pill carries out quality control not appeared in the newspapers as yet from more macroscopical angle.
Summary of the invention
The assay method that the purpose of this invention is to provide a kind of fingerprint pattern of compound salvia dropping pills, by this assay method, may command compound danshen dripping pills quality.
The present invention can implement by following step:
(a) preparation of compound danshen dripping pills solution: take by weighing compound danshen dripping pills, be dissolved in the ammoniacal liquor ultrasonic dissolution, filtering membrane, get the C-18 pillar that filtrate is crossed filler, washing fluid is abandoned in the flushing of methanol aqueous ammonia solution, wash with water again, abandon water lotion, use methanol-eluted fractions again, collect meoh eluate in volumetric flask, constant volume, centrifugal standby;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is a glacial acetic acid aqueous solution, and Mobile phase B is the glacial acetic acid acetonitrile solution; Detect wavelength 200～210nm;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains finger-print.
Preferred the present invention can implement by following step, and the methanol aqueous ammonia solution described in the described step (a) is 5～40% methyl alcohol, 0.5～10% ammonia spirits.
Further preferred the present invention can implement by following step: the preparation of compound danshen dripping pills solution in the described step (a): take by weighing compound danshen dripping pills, be dissolved in the ammoniacal liquor of 1～20mL1～8%, ultrasonic dissolution, cross 0.30～0.60 μ m filter membrane, get the C-18 pillar that 2～10mL filtrate is crossed filler, methyl alcohol 1～8% ammonia spirit flushing of 5～40mL8～35%, abandon washing fluid, wash with 5～35ml again, abandon water lotion, use methanol-eluted fractions again, collect meoh eluate in 1～15mL volumetric flask, constant volume, centrifugal standby;
Chromatographic condition in the described step (b): sample size 5～40 μ L; Detect wavelength 201～205nm; 20～40 ℃ of column temperatures.
Further preferred the present invention can implement by following step:
(a) preparation of compound danshen dripping pills need testing solution: take by weighing compound danshen dripping pills, be dissolved in 5～15mL2～6% ammoniacal liquor, ultrasonic dissolution, cross 0.35～0.55 μ m filter membrane, get the C-18 pillar that 2～10mL filtrate is crossed filler, methyl alcohol 2～6% ammonia spirits flushing of 10～30mL10～30%, abandon washing fluid, with 10～30ml washing, abandon water lotion again, use methanol-eluted fractions again, collect meoh eluate in 1～10mL volumetric flask, constant volume, centrifugal standby, sample size 15～25 μ L;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is 0.01% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.01% glacial acetic acid; The gradient elution program is as follows:
In the time of 0 minute, mobile phase A is that 80% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 20% 0.01% glacial acetic acid;
In the time of 15 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid;
In the time of 25 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid;
In the time of 40 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid;
In the time of 50 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid;
In the time of 65 minutes, mobile phase A is that 42% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 58% 0.01% glacial acetic acid;
In the time of 75 minutes, mobile phase A is that 25% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 75% 0.01% glacial acetic acid; Flow velocity 0.8mLmin
-1Detect wavelength 202～204nm; 28～32 ℃ of column temperatures;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains finger-print.
Best the present invention can implement by following step:
The preparation of compound danshen dripping pills solution in the described step (a): take by weighing compound danshen dripping pills 0.4～1.5g, be dissolved in 10mL4% ammoniacal liquor, ultrasonic dissolution, cross 0.45 μ m filter membrane, get the C-18 pillar that 5mL filtrate is crossed the 500mg filler, washing fluid is abandoned in the methyl alcohol 4% ammonia spirit flushing of 20mL20%, wash with 20ml again, abandon water lotion, use methanol-eluted fractions again, collect meoh eluate in the 5mL volumetric flask, constant volume, centrifugal standby;
Chromatographic condition in the described step (b): sample size 20 μ L; Flow velocity 0.8mLmin
-1Detect wavelength 203nm; 30 ℃ of column temperatures.
Among the present invention, the fingerprint pattern of compound salvia dropping pills effect of selecting for use methanol aqueous ammonia solution flushing to be obtained is better than the effect of the fingerprint pattern of compound salvia dropping pills that the water flushing obtained, and its washing fluid methanol aqueous ammonia solution all has effect preferably in the scope of 5～40% methyl alcohol 0.5～10% ammonia spirit; Methyl alcohol 1～8% ammonia spirit flushing of preferred 5～40mL8～35%; Further methyl alcohol 2～6% ammonia spirits of preferred 10～30mL10～30% flushing, the methyl alcohol 4% ammonia spirit flushing of best 20mL20%.
According to inventive method mensuration of the present invention is the finger-print of pseudo-ginseng chemical constitution in the compound danshen dripping pills, according to the method described above in the finger-print of Ce Dinging, pseudo-ginseng component chemical composition absorption peak has 14 in the compound danshen dripping pills, the absorption peak that wherein unimodal area surpasses total peak area 2% has 13, is respectively:
No. 1 peak, average retention time RT is 10.87min, and RSD is 0.31%, and peak area is 506.92, and RSD is 13.03%;
No. 2 peaks, average retention time RT is 12.12min, and RSD is 0.34%, and peak area is 2723.73, and RSD is 12.11%;
No. 3 peaks, average retention time RT is 20.34min, and RSD is 0.23%, and peak area is 1684.49, and RSD is 18.59%;
No. 5 peaks, average retention time RT is 23.29min, and RSD is 0.25%, and peak area is 565.78, and RSD is 13.80%;
No. 6 peaks, average retention time RT is 24.48min, and RSD is 0.28%, and peak area is 309.39, and RSD is 18.98%;
No. 7 peaks, average retention time RT is 29.00min, and RSD is 0.62%, and peak area is 345.32, and RSD is 14.28%;
No. 8 peaks, average retention time RT is 41.21min, and RSD is 0.25%, and peak area is 436.06, and RSD is 10.88%;
No. 9 peaks, average retention time RT is 42.94min, and RSD is 0.21%, and peak area is 665.38, and RSD is 11.82%;
No. 10 peaks, average retention time RT is 44.16min, and RSD is 0.21%, and peak area is 472.84, and RSD is 14.59%;
No. 11 peaks, average retention time RT is 45.68min, and RSD is 0.23%, and peak area is 947.39, and RSD is 16.06%;
No. 12 peaks, average retention time RT is 47.88min, and RSD is 0.25%, and peak area is 1547.80, and RSD is 13.25%;
No. 13 peaks, average retention time RT is 68.21min, and RSD is 0.13%, and peak area is 622.91, and RSD is 10.39%;
No. 14 peaks, average retention time RT is 69.43min, and RSD is 0.12%, and peak area is 820.24, and RSD is 12.94%;
In the described finger-print, pseudo-ginseng component chemical composition absorption peak has 14 in the compound danshen dripping pills, and the absorption peak that wherein unimodal area surpasses total peak area 5% has 7, is respectively:
No. 2 peaks, average retention time RT is 12.12min, and RSD is 0.34%, and peak area is 2723.73, and RSD is 12.11%;
No. 3 peaks, average retention time RT is 20.34min, and RSD is 0.23%, and peak area is 1684.49, and RSD is 18.59%;
No. 9 peaks, average retention time RT is 42.94min, and RSD is 0.21%, and peak area is 665.38, and RSD is 11.82%;
No. 11 peaks, average retention time RT is 45.68min, and RSD is 0.23%, and peak area is 947.39, and RSD is 16.06%;
No. 12 peaks, average retention time RT is 47.88min, and RSD is 0.25%, and peak area is 1547.80, and RSD is 13.25%;
No. 13 peaks, average retention time RT is 68.21min, and RSD is 0.13%, and peak area is 622.91, and RSD is 10.39%;
No. 14 peaks, average retention time RT is 69.43min, and RSD is 0.12%, and peak area is 820.24, and RSD is 12.94%;
In the described finger-print, pseudo-ginseng component chemical composition absorption peak has 14 in the compound danshen dripping pills, and the absorption peak that wherein unimodal area surpasses total peak area 10% has 3, is respectively:
No. 2 peaks, average retention time RT is 12.12min, and RSD is 0.34%, and peak area is 2723.73, and RSD is 12.11%;
No. 3 peaks, average retention time RT is 20.34min, and RSD is 0.23%, and peak area is 1684.49, and RSD is 18.59%;
No. 12 peaks, average retention time RT is 47.88min, and RSD is 0.25%, and peak area is 1547.80, and RSD is 13.25%.
In the assay method of above-mentioned fingerprint pattern of compound salvia dropping pills, its configuration proportion that is used for the mobile phase A solution of gradient elution is to prepare by volume, and the configuration proportion of Mobile phase B solution is to prepare by volume.
The present invention measures pseudo-ginseng chemical composition in pseudo-ginseng, the compound danshen dripping pills intermediate, and method is as follows:
Pseudo-ginseng chemical composition high-efficiency liquid-phase fingerprint acquisition methods in the pseudo-ginseng, method is as follows:
The preparation of pseudo-ginseng test sample: get it filled material in Backflow bottle, add the distilled water refluxing extraction, filter, add distilled water in the filter residue, reflux, merging filtrate is dissolved in surely, and centrifugal filtration is standby;
The accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains the finger-print of pseudo-ginseng component chemical composition.
Pseudo-ginseng chemical composition high-efficiency liquid-phase fingerprint acquisition methods in the preferred pseudo-ginseng, method is as follows:
The preparation of pseudo-ginseng test sample: take by weighing medicinal material 2.5g in Backflow bottle, add distilled water 50mL, refluxed 1.5 hours, filter, add 50mL distilled water in the filter residue, refluxed 1 hour, merging filtrate is dissolved in the 100mL measuring bottle surely, and centrifugal filtration is standby; Parallel 2 parts, sample size 10 μ L;
The accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains the finger-print of pseudo-ginseng component chemical composition;
In the described finger-print, the component chemical composition absorption peak of pseudo-ginseng test sample has 5, and the absorption peak that wherein unimodal area surpasses total peak area 2% has 5, is respectively:
No. 1 peak, average retention time RT is 10.81min, and RSD is 0.06%, and peak area is 539.57, and RSD is 17.73%;
No. 2 peaks, average retention time RT is 12.03min, and RSD is 0.02%, and peak area is 2482.50, and RSD is 8.74%;
No. 3 peaks, average retention time RT is 20.16min, and RSD is 0.11%, and peak area is 1514.5, and RSD is 12.02%;
No. 4 peaks, average retention time RT is 23.04min, and RSD is 0.07%, and peak area is 250.63, and RSD is 18.25%;
No. 5 peaks, average retention time RT is 28.42min, and RSD is 0.27%, and peak area is 356.20, and RSD is 15.30%.
In the described finger-print, the component chemical composition absorption peak of pseudo-ginseng test sample has 5, and the absorption peak that wherein unimodal area surpasses total peak area 5% has 4, is respectively:
No. 1 peak, average retention time RT is 10.81min, and RSD is 0.06%, and peak area is 539.57, and RSD is 17.73%;
No. 2 peaks, average retention time RT is 12.03min, and RSD is 0.02%, and peak area is 2482.50, and RSD is 8.74%;
No. 3 peaks, average retention time RT is 20.16min, and RSD is 0.11%, and peak area is 1514.5, and RSD is 12.02%;
No. 5 peaks, average retention time RT is 28.42min, and RSD is 0.27%, and peak area is 356.20, and RSD is 15.30%.
In the described finger-print, the component chemical composition absorption peak of pseudo-ginseng test sample has 5, and the absorption peak that wherein unimodal area surpasses total peak area 10% has 3, is respectively:
No. 1 peak, average retention time RT is 10.81min, and RSD is 0.06%, and peak area is 539.57, and RSD is 17.73%;
No. 2 peaks, average retention time RT is 12.03min, and RSD is 0.02%, and peak area is 2482.50, and RSD is 8.74%;
No. 3 peaks, average retention time RT is 20.16min, and RSD is 0.11%, and peak area is 1514.5, and RSD is 12.02%.
Pseudo-ginseng chemical composition high-efficiency liquid-phase fingerprint acquisition methods in the compound danshen dripping pills intermediate, method is as follows:
The preparation of compound danshen dripping pills intermediate test sample: take by weighing the compound danshen dripping pills intermediate, be dissolved in ammoniacal liquor, ultrasonic dissolution, filtering membrane, get filtrate and cross the C-18 pillar, wash with the ammonia spirit of methyl alcohol earlier, wash with water again, collect meoh eluate in volumetric flask, constant volume, centrifugal standby;
Pseudo-ginseng chemical composition high-efficiency liquid-phase fingerprint acquisition methods in the preferred compound danshen dripping pills intermediate, method is as follows:
The preparation of compound danshen dripping pills intermediate test sample: take by weighing compound danshen dripping pills intermediate 0.1～0.5g, be dissolved in 10mL4% ammoniacal liquor, ultrasonic dissolution, cross 0.45 μ m filter membrane, get 5mL filtrate and cross the C-18 pillar, elder generation washes with the ammonia spirit of the methyl alcohol 4% of 15ml20%, wash with 20mL again, collect meoh eluate in the 5mL volumetric flask, constant volume, centrifugal standby; Parallel 2 parts, sample size 20 μ L;
The accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains the finger-print of pseudo-ginseng component chemical composition;
Pseudo-ginseng component chemical composition absorption peak has 12 in the compound danshen dripping pills intermediate, and the absorption peak that wherein unimodal area surpasses total peak area 2% has 11, is respectively:
No. 1 peak, average retention time RT is 10.85min, and RSD is 0.08%, and peak area is 557.29, and RSD is 15.67%;
No. 2 peaks, average retention time RT is 12.10min, and RSD is 0.06%, and peak area is 2728.83, and RSD is 16.66%;
No. 3 peaks, average retention time RT is 20.33min, and RSD is 0.08%, and peak area is 3704.31, and RSD is 14.15%;
No. 4 peaks, average retention time RT is 23.86min, and RSD is 0.07%, and peak area is 272.69, and RSD is 12.54%;
No. 5 peaks, average retention time RT is 23.28min, and RSD is 0.08%, and peak area is 369.12, and RSD is 21.52%;
No. 7 peaks, average retention time RT is 28.82min, and RSD is 1.36%, and peak area is 289.16, and RSD is 17.71%;
No. 8 peaks, average retention time RT is 42.93min, and RSD is 0.07%, and peak area is 239.14, and RSD is 18.84%;
No. 9 peaks, average retention time RT is 45.66min, and RSD is 0.06%, and peak area is 445.17, and RSD is 19.56%;
No. 10 peaks, average retention time RT is 47.85min, and RSD is 0.06%, and peak area is 723.59, and RSD is 14.24%;
No. 11 peaks, average retention time RT is 68.17min, and RSD is 0.03%, and peak area is 407.66, and RSD is 10.86%;
No. 12 peaks, average retention time RT is 69.4min, and RSD is 0.03%, and peak area is 476.31, and RSD is 15.46%.
Pseudo-ginseng component chemical composition absorption peak has 12 in the compound danshen dripping pills intermediate, and the absorption peak that wherein unimodal area surpasses total peak area 5% has 4, is respectively:
No. 1 peak, average retention time RT is 10.85min, and RSD is 0.08%, and peak area is 557.29, and RSD is 15.67%;
No. 2 peaks, average retention time RT is 12.10min, and RSD is 0.06%, and peak area is 2728.83, and RSD is 16.66%;
No. 3 peaks, average retention time RT is 20.33min, and RSD is 0.08%, and peak area is 3704.31, and RSD is 14.15%;
No. 10 peaks, average retention time RT is 47.85min, and RSD is 0.06%, and peak area is 723.59, and RSD is 14.24%.
Pseudo-ginseng component chemical composition absorption peak has 12 in the compound danshen dripping pills intermediate, and the absorption peak that wherein unimodal area surpasses total peak area 10% has 2, is respectively:
No. 2 peaks, average retention time RT is 12.10min, and RSD is 0.06%, and peak area is 2728.83, and RSD is 16.66%;
No. 3 peaks, average retention time RT is 20.33min, and RSD is 0.08%, and peak area is 3704.31, and RSD is 14.15%;
No. 10 peaks, average retention time RT is 47.85min, and RSD is 0.06%, and peak area is 723.59, and RSD is 14.24%.
The present invention finds by the finger-print of measuring compound danshen dripping pills, compound danshen dripping pills intermediate, pseudo-ginseng, absorption peak quantity and incomplete same among the three, illustrate finger-print that adopt to measure the compound danshen dripping pills product as the compound danshen dripping pills quality control standard than separately with the finger-print of pseudo-ginseng composition as quality standard more can objectively reflect the inherent composition of product having or not with and content, with the quality of comprehensive control product.
Advantage of the present invention is as follows:
(1). with main effective constituent pseudo-ginseng chemical composition in the compound danshen dripping pills is the HPLC finger-print that index is set up, and is representing the most of pharmacologically active of compound danshen dripping pills, can characterize the quality of compound danshen dripping pills effectively.Thereby realize the chemical constitution of compound danshen dripping pills maximum possible is detected, help monitoring in all directions traditional Chinese medicine quality.
(2). do as a wholely to treat with each effective constituent fingerprint graph of pseudo-ginseng in the compound danshen dripping pills, pay attention to each front and back that constitute fingerprint characteristic peak order and mutual relationship, pay attention to whole facial feature, both avoided judging the one-sidedness of compound danshen dripping pills total quality that having reduced again was the possibility of the artificial processing of requisite quality because of only measuring one, two chemical constitution.The present invention provides new reference standard for quality complete, that accurately estimate compound danshen dripping pills, will be that the quality and the curative effect of principal ingredient prescribed preparation contributes with the red sage root, pseudo-ginseng for improving compound danshen dripping pills and other.
(3). the present invention has that method is easy, stable, precision is high, favorable reproducibility, the characteristics that are easy to grasp.
The invention provides a kind of detection method of compound danshen dripping pills, the present invention is by the resulting practicable method of a large amount of experiments, has repeatability.The repeatability of red sage root chemical composition high-efficiency liquid-phase fingerprint by red sage root chemical composition high-efficiency liquid-phase fingerprint, compound danshen dripping pills intermediate in high performance liquid chromatogram, the compound danshen dripping pills that obtained under the same test condition and red rooted salvia finger-print is good in the present invention, measure good reproducibility, therefore the fingerprint pattern of compound salvia dropping pills that can pass through the said determination method is set up is as standard finger-print, its assay method can be used as the standard finger-print that contains the red sage root, notoginseng preparations effective constituent determination, to differentiate its effective constituent.
To those skilled in the art, technology contents disclosed according to the present invention, those skilled in the art will very clear other embodiment of the present invention, and the embodiment of the invention is only as example.Under the situation of not violating purport of the present invention and scope, can carry out various changes and improvements to the present invention.For example, use the different measurement result that detecting instrument obtained possibilities different, but as long as use method of quality control of the present invention, all within protection domain of the present invention.
Description of drawings
Provide the contrast of the pseudo-ginseng chemical composition HPLC finger-print of pseudo-ginseng HPLC finger-print, separate sources in pseudo-ginseng HPLC finger-print, the compound danshen dripping pills in pseudo-ginseng HPLC finger-print, the compound danshen dripping pills intermediate below, be intended to further specify the present invention, but the present invention do not limited.
Fig. 1 pseudo-ginseng HPLC finger-print
Pseudo-ginseng HPLC finger-print in Fig. 2 compound danshen dripping pills intermediate
Pseudo-ginseng HPLC finger-print in Fig. 3 compound danshen dripping pills
The contrast of the pseudo-ginseng chemical composition HPLC finger-print of Fig. 4 separate sources
Wherein: 1 is compound danshen dripping pills
2 is the compound danshen dripping pills intermediate
3 is pseudo-ginseng
Embodiment
The embodiment that enumerates the preparation aspect below further describes the present invention, and this embodiment only is used to the present invention is described and the present invention is not limited.
Embodiment one (preparation example)
Take by weighing red sage root 41.06g, pseudo-ginseng 8.03g, the boiling secondary, the first time, 4 times of water gagings were 2 hours, the second time, 3 times of water gagings were 1 hour, filter, filtrate merges, when being concentrated into proportion and being 1.19-1.20 (75 ± 1 ℃), adding concentration is that 90% left and right sides ethanol to ethanol content is 65% (20 ℃), left standstill 12 hours, separation of supernatant reclaims ethanol, being concentrated into relative density of medicine liquid is 1.37 (55-60 ℃), gets Salvia miltiorrhiza and Panax notoginseng medicinal extract.Get above-mentioned Salvia miltiorrhiza and Panax notoginseng medicinal extract and borneol 0.46g, evenly be heated to 85 ℃ of temperature, change material after 80 minutes, move in the dropping-pill machine jar that jar temperature remains on 86 ℃ with polyglycol-6000 18g is mixed.In medicine liquid droplet to the 8 ℃ whiteruss, take out dripping pill, oil removing, screen cloth selects ball, promptly.
Embodiment two (preparation example)
Take by weighing red sage root 59.36g, pseudo-ginseng 6.38g, add the sal tartari of medicinal material total amount 1.0%, decoct secondary, the first time, 4 times of water gagings were 2.5 hours, and the second time, 3 times of water gagings were 1.5 hours, filtered, filtrate merges, when being concentrated into proportion and being 1.19-1.20 (75 ± 1 ℃), adding concentration is that 85% left and right sides ethanol to ethanol content is 70% (20 ℃), leaves standstill 10 hours, separation of supernatant, reclaim ethanol, being concentrated into relative density of medicine liquid is 1.35 (55-60 ℃), gets Salvia miltiorrhiza and Panax notoginseng medicinal extract.Get above-mentioned Salvia miltiorrhiza and Panax notoginseng medicinal extract and borneol 0.34g, evenly be heated to 89 ℃ of temperature, change material after 100 minutes, move in the dropping-pill machine jar that jar temperature remains on 85 ℃ with polyglycol-6000 23g is mixed.In medicine liquid droplet to the 8 ℃ methyl-silicone oil, take out dripping pill, oil removing, screen cloth selects ball, promptly.
Embodiment three (preparation example)
Take by weighing red sage root 31.12g, pseudo-ginseng 9.21g, the boiling secondary, the first time, 4 times of water gagings were 1.5 hours, the second time, 3 times of water gagings were 1.5 hours, filter, filtrate merges, when being concentrated into proportion and being 1.19-1.20 (75 ± 1 ℃), adding concentration is that 88% left and right sides ethanol to ethanol content is 66% (20 ℃), left standstill 10 hours, separation of supernatant reclaims ethanol, being concentrated into relative density of medicine liquid is 1.40 (55-60 ℃), gets Salvia miltiorrhiza and Panax notoginseng medicinal extract.Get above-mentioned Salvia miltiorrhiza and Panax notoginseng medicinal extract and borneol 0.50g, sweet mellow wine 90g, mosatil 15g and distilled water 15ml, behind the said components mixing, powder-injection is made in freeze drying.
Embodiment four (preparation example)
Take by weighing red sage root 116.35g, pseudo-ginseng 58.21g, add the sodium bicarbonate of medicinal material total amount 2.0%, decoct secondary, the first time, 4 times of water gagings were 2 hours, and the second time, 3 times of water gagings were 1.5 hours, filtered, filtrate merges, when being concentrated into proportion and being 1.19-1.20 (75 ± 1 ℃), adding concentration is that 88% left and right sides ethanol to ethanol content is 66% (20 ℃), leaves standstill 10 hours, separation of supernatant, reclaim ethanol, being concentrated into relative density of medicine liquid is 1.40 (55-60 ℃), gets Salvia miltiorrhiza and Panax notoginseng medicinal extract.Get above-mentioned Salvia miltiorrhiza and Panax notoginseng medicinal extract and dalbergia heartwood oil 1.8g, mix, add 3% polyvidone ethanolic solution system softwood, cross 18 mesh sieve system particles with the 40g microcrystalline cellulose, 60 ℃ of dryings 35 minutes, whole grain, adding 4g talcum powder, mixing fills in capsule, promptly.
Embodiment five (preparation example)
Take by weighing red sage root 116.35g, pseudo-ginseng 58.21g, the boiling secondary, the first time, 4 times of water gagings were 2 hours, the second time, 3 times of water gagings were 1.5 hours, filter, filtrate merges, when being concentrated into proportion and being 1.19-1.20 (75 ± 1 ℃), adding concentration is that 88% left and right sides ethanol to ethanol content is 66% (20 ℃), left standstill 10 hours, separation of supernatant reclaims ethanol, being concentrated into relative density of medicine liquid is 1.40 (55-60 ℃), gets Salvia miltiorrhiza and Panax notoginseng medicinal extract.Get above-mentioned Salvia miltiorrhiza and Panax notoginseng medicinal extract and borneol 0.9g, mix with microcrystalline cellulose 120g, HPMC 40g, xylitol 5g, dolomol 2g, compressing tablet, promptly.
Embodiment six (preparation example)
Take by weighing red sage root 140.35g, pseudo-ginseng 36.42g, add the sodium bicarbonate of medicinal material total amount 2.5%, decoct secondary, the first time, 4 times of water gagings were 2 hours, and the second time, 3 times of water gagings were 1.5 hours, filtered, filtrate merges, when being concentrated into proportion and being 1.19-1.20 (75 ± 1 ℃), adding concentration is that 90% left and right sides ethanol to ethanol content is 65% (20 ℃), leaves standstill 8 hours, separation of supernatant, reclaim ethanol, being concentrated into relative density of medicine liquid is 1.35 (55-60 ℃), gets Salvia miltiorrhiza and Panax notoginseng medicinal extract.Get above-mentioned Salvia miltiorrhiza and Panax notoginseng medicinal extract and borneol 1.0g, mix, add 3% polyvidone ethanolic solution system softwood, cross 18 mesh sieve system particles with the 46g microcrystalline cellulose, 60 ℃ of dryings 30 minutes, whole grain, adding 4g talcum powder, mixing, compressing tablet, promptly.
Embodiment seven (compound danshen dripping pills pseudo-ginseng component finger-print test example)
1, instrument and reagent
Instrument: adopt Agilent 1100 liquid chromatographies, comprise quaternary pump, online degasser, automatic sampler, DAD detecting device, column oven, Chemstation workstation; BS210S electronic balance (1/10
-4G) (Beijing Sai Duolisi company), METTLERAE240 electronic balance (1/10
-4G or 1/10
-5G) (plum Teller-Tuo benefit (Shanghai) Co., Ltd.), LD4-2 hydro-extractor (4000r/min) (Beijing Medical Centrifugal Machine Factory), digital display thermostat water bath (the long wind in Tianjin company limited), RE-52AA rotary evaporator (Shanghai Yarong Biochemical Instrument Plant), SHE-(III) circulation ability of swimming vacuum pump (Gongyi Ying Yu gives magnificent instrument plant), KQ-250B ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), HENGAO T﹠amp; D filtrator (HENGGAO T﹠amp; D), synthon filtering membrane (aperture 0.45 μ m) (going up Haixing County inferior scavenging material factory); C
18(filler is C to pillar
18H
17The liquid chromatography stationary phase, 50～100 orders, chemical reagent two factories in Tianjin produce.Dry column-packing 600mg, column internal diameter 1cm).
Reagent: acetonitrile (chromatographically pure, U.S. Merck company), acetic acid (top grade is pure), Wahaha pure water.
2, test sample preparation
The preparation of compound danshen dripping pills test sample: take by weighing each batch compound danshen dripping pills 1.0g among the embodiment one, be dissolved in 10ml4% ammoniacal liquor, ultrasonic dissolution, cross 0.45 μ m filter membrane, get 5ml filtrate and cross C-18 pillar (500mg filler), the 20ml washing is collected meoh eluate in the 5ml volumetric flask, constant volume, centrifugal standby.Parallel 2 parts, sample size 20 μ l.
The preparation of compound danshen dripping pills intermediate test sample: take by weighing each batch Salvia miltiorrhiza and Panax notoginseng medicinal extract 0.2g among the embodiment one, be dissolved in 10ml4% ammoniacal liquor, ultrasonic dissolution, cross 0.45 μ m filter membrane, get 5ml filtrate and cross the C-18 pillar, elder generation washes with the ammonia spirit of the methyl alcohol 4% of 15ml20%, wash with 20ml again, collect meoh eluate in the 5ml volumetric flask, constant volume, centrifugal standby.Parallel 2 parts, sample size 20 μ l.
The preparation of pseudo-ginseng test sample: take by weighing medicinal material 2.5g in Backflow bottle, add distilled water 50ml, refluxed 1.5 hours, filter, add 50ml distilled water in the filter residue, refluxed 1 hour, merging filtrate is dissolved in the 100ml measuring bottle surely, and centrifugal filtration is standby.Parallel 2 parts, sample size 10 μ l.
3, HPLC analysis condition
Agilent SB-C
18Analytical column (4.6mm * 250mm, Zorbax SB USCL010304); Moving phase: A is 0.01% (V/V) glacial acetic acid aqueous solution mutually, and B is mutually for containing the acetonitrile solution of 0.01% glacial acetic acid.Flow velocity 0.8mlmin
-1Detect wavelength 203nm; 30 ℃ of column temperatures.
Gradient elution program such as following table:
Time???????????????A：0.01％HAC-H2O(％)(v/v)????????B：0.01％HAC-CH3CN(％)(v/v)
0?????????????????????????????80????????????????????????????????20
15????????????????????????????65????????????????????????????????35
25????????????????????????????65????????????????????????????????35
40????????????????????????????57????????????????????????????????43
50????????????????????????????57????????????????????????????????43
65????????????????????????????42????????????????????????????????58
75????????????????????????????25????????????????????????????????75
Compound danshen dripping pills pseudo-ginseng finger-print composition title:
| Peak 1- | Peak 2- | Peak 3- | Peak 4- | Peak 5- | Peak 6- | Peak 7- |
| Panax Notoginseng saponin R 1 | Ginsenoside R e+R g1 | Ginsenoside R b1 | Panax Notoginseng saponin R 2 | Ginsenoside R h1 | Ginsenoside Rh liso. (F1) | Ginsenoside R d |
| Peak 8- | Peak 9- | Peak 10- | Peak 11- | Peak 12- | Peak 13- | Peak 14- |
| Ginsenoside Ng-R2-H2O | Ginsenoside Ng-R2-H2O | Ginsenoside R g6/F 4 | Ginsenoside R k3/R h4(R k3) | Ginsenoside R k3/R h4(R h4) | Ginsenoside R k1/R g5(R k1) | Ginsenoside R k1/R g5(R g5) |
Amount to reply side's Radix Salviae Miltiorrhizae extractum surplus 200 is carried out the red sage root, pseudo-ginseng fingerprint map analyzing respectively, its similarity is all more than 90%.Now gather the retention time of collection of illustrative plates, the mean value and the RSD value of peak area as follows:
Pseudo-ginseng finger-print part
| Peak number | Average retention time | The RSD% of retention time | Average peak area | The RSD% of peak area | The unimodal number percent that accounts for total peak area |
| ????1 | ????10.85 | ????0.08 | ??557.29 | ????15.67 | ????5.36％ |
| ????2 | ????12.1 | ????0.06 | ??2728.83 | ????16.66 | ????26.23％ |
| ????3 | ????20.33 | ????0.08 | ??3704.31 | ????14.15 | ????35.61％ |
| ????4 | ????20.86 | ????0.07 | ??272.69 | ????12.54 | ????2.62％ |
| ????5 | ????23.28 | ????0.08 | ??369.12 | ????21.52 | ????3.55％ |
| ????6 | ????24.48 | ????0.08 | ??190.23 | ????17.45 | ????1.83％ |
| ????7 | ????28.82 | ????1.36 | ??289.16 | ????17.71 | ????2.78％ |
| ????8 | ????42.93 | ????0.07 | ??239.14 | ????18.84 | ????2.30％ |
| ????9 | ????45.66 | ????0.06 | ??445.17 | ????19.56 | ????4.28％ |
| ????10 | ????47.85 | ????0.06 | ??723.59 | ????14.24 | ????6.96％ |
| ????11 | ????68.17 | ????0.03 | ??407.66 | ????10.86 | ????3.92％ |
| ????12 | ????69.4 | ????0.03 | ??476.31 | ????15.46 | ????4.58％ |
Amount to reply side's Danshen Root dropping ball surplus 200 is carried out the red sage root, pseudo-ginseng fingerprint map analyzing respectively, its similarity is all more than 90%.Now gather the retention time of collection of illustrative plates, the mean value and the RSD value of peak area as follows:
Pseudo-ginseng finger-print part
| Peak number | Average retention time | Retention time RSD% | Average peak area | Peak area RSD% | The unimodal number percent that accounts for total peak area |
| ????1 | ????10.87 | ????0.45 | ??506.92 | ????13.03 | ????4.27％ |
| ????2 | ????12.12 | ????0.34 | ??2723.73 | ????12.11 | ????22.93％ |
| ????3 | ????20.34 | ????0.23 | ??1684.49 | ????18.95 | ????14.18％ |
| ????4 | ????20.86 | ????0.16 | ??231.84 | ????18.99 | ????1.95％ |
| ????5 | ????23.29 | ????0.25 | ??565.78 | ????13.8 | ????4.76％ |
| ????6 | ????24.48 | ????0.28 | ??309.39 | ????18.98 | ????2.60％ |
| ????7 | ????29 | ????0.62 | ??345.32 | ????14.28 | ????2.91％ |
| ????8 | ????41.21 | ????0.25 | ??436.06 | ????10.88 | ????3.67％ |
| ????9 | ????42.94 | ????0.21 | ??665.38 | ????11.82 | ????5.60％ |
| ????10 | ????44.16 | ????0.21 | ??472.84 | ????14.59 | ????3.98％ |
| ????11 | ????45.68 | ????0.23 | ??947.39 | ????16.06 | ????7.97％ |
| ????12 | ????47.88 | ????0.25 | ??1547.8 | ????13.25 | ????13.03％ |
| ????13 | ????68.21 | ????0.13 | ??622.91 | ????10.39 | ????5.24％ |
| ????14 | ????69.43 | ????0.12 | ??820.24 | ????12.93 | ????6.90％ |
The assay method of embodiment eight fingerprint pattern of compound salvia dropping pills
(a) preparation of compound danshen dripping pills solution: take by weighing compound danshen dripping pills, be dissolved in the ammoniacal liquor of 20mL5% ultrasonic dissolution, cross 0.30 μ m filter membrane, get the C-18 pillar that 10mL filtrate is crossed filler, the methyl alcohol 4% ammonia spirit flushing of 40mL10%, abandon washing fluid, with the 35ml washing, abandon water lotion again, use methanol-eluted fractions again, collect meoh eluate in the 15mL volumetric flask, constant volume, centrifugal standby, sample size 5 μ L;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is 0.01% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.01% glacial acetic acid; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 80% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 20% 0.01% glacial acetic acid; In the time of 15 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid; In the time of 25 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid; In the time of 40 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid; In the time of 65 minutes, mobile phase A is that 42% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 58% 0.01% glacial acetic acid; In the time of 75 minutes, mobile phase A is that 25% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 75% 0.01% glacial acetic acid; Flow velocity 0.8mLmin
-1Detect wavelength 203nm; 30 ℃ of column temperatures;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains the finger-print of pseudo-ginseng component chemical composition.
The assay method of embodiment nine fingerprint pattern of compound salvia dropping pills
(a) preparation of compound danshen dripping pills need testing solution: take by weighing compound danshen dripping pills, be dissolved in 15mL6% ammoniacal liquor, ultrasonic dissolution, cross 0.55 μ m filter membrane, get the C-18 pillar that 10mL filtrate is crossed filler, the methyl alcohol 6% ammonia spirit flushing of 30mL30%, abandon washing fluid, with the 30ml washing, abandon water lotion again, use methanol-eluted fractions again, collect meoh eluate 5mL volumetric flask, constant volume, centrifugal standby, sample size 15 μ L;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is 0.01% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.01% glacial acetic acid; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 80% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 20% 0.01% glacial acetic acid; In the time of 15 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid; In the time of 25 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid; In the time of 40 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid; In the time of 65 minutes, mobile phase A is that 42% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 58% 0.01% glacial acetic acid; In the time of 75 minutes, mobile phase A is that 25% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 75% 0.01% glacial acetic acid; Flow velocity 0.8mLmin
-1Detect wavelength 203nm; 30 ℃ of column temperatures;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains the finger-print of pseudo-ginseng component chemical composition.
The assay method of embodiment ten fingerprint pattern of compound salvia dropping pills
(a) preparation of compound danshen dripping pills need testing solution: take by weighing compound danshen dripping pills 1.0g, be dissolved in 10mL4% ammoniacal liquor, ultrasonic dissolution, cross 0.45 μ m filter membrane, get the C-18 pillar that 5mL filtrate is crossed the 500mg filler, the methyl alcohol 4% ammonia spirit flushing of 20mL20%, abandon washing fluid, with the 20ml washing, abandon water lotion again, use methanol-eluted fractions again, collect meoh eluate in the 5mL volumetric flask, constant volume, centrifugal standby, parallel 2 parts, sample size 20 μ L;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is 0.01% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.01% glacial acetic acid; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 80% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 20% 0.01% glacial acetic acid; In the time of 15 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid; In the time of 25 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid; In the time of 40 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid; In the time of 65 minutes, mobile phase A is that 42% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 58% 0.01% glacial acetic acid; In the time of 75 minutes, mobile phase A is that 25% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 75% 0.01% glacial acetic acid; Flow velocity 0.8mLmin
-1Detect wavelength 203nm; 30 ℃ of column temperatures;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains the finger-print of pseudo-ginseng component chemical composition;
In the described finger-print, pseudo-ginseng component chemical composition absorption peak has 14 in the compound danshen dripping pills, and the absorption peak that wherein unimodal area surpasses total peak area 2% has 13, they are that No. 1 average retention time RT in peak is 10.87min, RSD is 0.31%, and peak area is 506.92, and RSD is 13.03%; No. 2 the average retention time RT in peak is 12.12min, and RSD is 0.34%, and peak area is 2723.73, and RSD is 12.11%; No. 3 the average retention time RT in peak is 20.34min, and RSD is 0.23%, and peak area is 1684.49, and RSD is 18.59%; No. 5 the average retention time RT in peak is 23.29min, and RSD is 0.25%, and peak area is 565.78, and RSD is 13.80%; No. 6 the average retention time RT in peak is 24.48min, and RSD is 0.28%, and peak area is 309.39, and RSD is 18.98%; No. 7 the average retention time RT in peak is 29.00min, and RSD is 0.62%, and peak area is 345.32, and RSD is 14.28%; No. 8 the average retention time RT in peak is 41.21min, and RSD is 0.25%, and peak area is 436.06, and RSD is 10.88%; No. 9 the average retention time RT in peak is 42.94min, and RSD is 0.21%, and peak area is 665.38, and RSD is 11.82%; No. 10 the average retention time RT in peak is 44.16min, and RSD is 0.21%, and peak area is 472.84, and RSD is 14.59%; No. 11 the average retention time RT in peak is 45.68min, and RSD is 0.23%, and peak area is 947.39, and RSD is 16.06%; No. 12 the average retention time RT in peak is 47.88min, and RSD is 0.25%, and peak area is 1547.80, and RSD is 13.25%; No. 13 the average retention time RT in peak is 68.21min, and RSD is 0.13%, and peak area is 622.91, and RSD is 10.39%; No. 14 the average retention time RT in peak is 69.43min, and RSD is 0.12%, and peak area is 820.24, and RSD is 12.94%;
In the described finger-print, pseudo-ginseng component chemical composition absorption peak has 14 in the compound danshen dripping pills, and the absorption peak that wherein unimodal area surpasses total peak area 5% has 7, they are that No. 2 average retention time RT in peak are 12.12min, RSD is 0.34%, and peak area is 2723.73, and RSD is 12.11%; No. 3 the average retention time RT in peak is 20.34min, and RSD is 0.23%, and peak area is 1684.49, and RSD is 18.59%; No. 9 the average retention time RT in peak is 42.94min, and RSD is 0.21%, and peak area is 665.38, and RSD is 11.82%; No. 11 the average retention time RT in peak is 45.68min, and RSD is 0.23%, and peak area is 947.39, and RSD is 16.06%; No. 12 the average retention time RT in peak is 47.88min, and RSD is 0.25%, and peak area is 1547.80, and RSD is 13.25%; No. 13 the average retention time RT in peak is 68.21min, and RSD is 0.13%, and peak area is 622.91, and RSD is 10.39%; No. 14 the average retention time RT in peak is 69.43min, and RSD is 0.12%, and peak area is 820.24, and RSD is 12.94%;
In the described finger-print, pseudo-ginseng component chemical composition absorption peak has 14 in the compound danshen dripping pills, and the absorption peak that wherein unimodal area surpasses total peak area 10% has 3, they are that No. 2 average retention time RT in peak are 12.12min, RSD is 0.34%, and peak area is 2723.73, and RSD is 12.11%; No. 3 the average retention time RT in peak is 20.34min, and RSD is 0.23%, and peak area is 1684.49, and RSD is 18.59%; No. 12 the average retention time RT in peak is 47.88min, and RSD is 0.25%, and peak area is 1547.80, and RSD is 13.25%.
Claims (6)
1, a kind of assay method of fingerprint pattern of compound salvia dropping pills, its feature comprises the steps:
(a) preparation of compound danshen dripping pills solution: take by weighing compound danshen dripping pills, be dissolved in the ammoniacal liquor ultrasonic dissolution, filtering membrane, get the C-18 pillar that filtrate is crossed filler, washing fluid is abandoned in the flushing of methanol aqueous ammonia solution, wash with water again, abandon water lotion, use methanol-eluted fractions again, collect meoh eluate in volumetric flask, constant volume, centrifugal standby;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is a glacial acetic acid aqueous solution, and Mobile phase B is the glacial acetic acid acetonitrile solution; Detect wavelength 200～210nm;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains finger-print.
2, the assay method of fingerprint pattern of compound salvia dropping pills as claimed in claim 1 is characterized in that: the methanol aqueous ammonia solution described in the described step (a) is 5～40% methyl alcohol, 0.5～10% ammonia spirits.
3, the assay method of fingerprint pattern of compound salvia dropping pills as claimed in claim 1 is characterized in that:
The preparation of compound danshen dripping pills solution in the described step (a): take by weighing compound danshen dripping pills, be dissolved in the ammoniacal liquor of 1～20mL1～8% ultrasonic dissolution, cross 0.30～0.60 μ m filter membrane, get the C-18 pillar that 2～10mL filtrate is crossed filler, washing fluid is abandoned in methyl alcohol 1～8% ammonia spirit flushing of 5～40mL8～35%, wash with 5～35ml again, abandon water lotion, use methanol-eluted fractions again, collect meoh eluate in 1～15mL volumetric flask, constant volume, centrifugal standby;
Chromatographic condition in the described step (b): sample size 5～40 μ L; Detect wavelength 201～205nm; 20～40 ℃ of column temperatures.
4, the assay method of fingerprint pattern of compound salvia dropping pills as claimed in claim 1 is characterized in that:
(a) preparation of compound danshen dripping pills need testing solution: take by weighing compound danshen dripping pills, be dissolved in 5～15mL2～6% ammoniacal liquor, ultrasonic dissolution, cross 0.35～0.55 μ m filter membrane, get the C-18 pillar that 2～10mL filtrate is crossed filler, methyl alcohol 2～6% ammonia spirits flushing of 10～30mL10～30%, abandon washing fluid, with 10～30ml washing, abandon water lotion again, use methanol-eluted fractions again, collect meoh eluate in 1～10mL volumetric flask, constant volume, centrifugal standby, sample size 15～25 μ L;
(b) chromatographic condition: chromatographic column is that octadecylsilane chemically bonded silica is a filler; Adopt gradient elution, mobile phase A is 0.01% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.01% glacial acetic acid; The gradient elution program is as follows:
In the time of 0 minute, mobile phase A is that 80% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 20% 0.01% glacial acetic acid;
In the time of 15 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid;
In the time of 25 minutes, mobile phase A is that 65% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 35% 0.01% glacial acetic acid;
In the time of 40 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid;
In the time of 50 minutes, mobile phase A is that 57% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 43% 0.01% glacial acetic acid;
In the time of 65 minutes, mobile phase A is that 42% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 58% 0.01% glacial acetic acid;
In the time of 75 minutes, mobile phase A is that 25% 0.01% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 75% 0.01% glacial acetic acid; Flow velocity 0.8mLmin
-1Detect wavelength 202～204nm; 28～32 ℃ of column temperatures;
(c) measure: the accurate need testing solution of drawing injects liquid chromatograph, according to high effective liquid chromatography for measuring, obtains finger-print.
5, the assay method of fingerprint pattern of compound salvia dropping pills as claimed in claim 1 is characterized in that:
The preparation of compound danshen dripping pills solution in the described step (a): take by weighing compound danshen dripping pills 0.4～1.5g, be dissolved in 10mL4% ammoniacal liquor, ultrasonic dissolution, cross 0.45 μ m filter membrane, get the C-18 pillar that 5mL filtrate is crossed the 500mg filler, washing fluid is abandoned in the methyl alcohol 4% ammonia spirit flushing of 20mL20%, wash with 20ml again, abandon water lotion, use methanol-eluted fractions again, collect meoh eluate in the 5mL volumetric flask, constant volume, centrifugal standby;
Chromatographic condition in the described step (b): sample size 20 μ L; Flow velocity 0.8mLmin
-1Detect wavelength 203nm; 30 ℃ of column temperatures.
6, as the assay method of claim 1,2 or 3 described fingerprint pattern of compound salvia dropping pills, the configuration proportion that it is characterized in that mobile phase A solution is to prepare by volume, and the configuration proportion of Mobile phase B solution is to prepare by volume.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100552648A CN100442049C (en) | 2004-03-17 | 2005-03-17 | Method for measuring fingerprint pattern of compound salvia dropping pills |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200410018757.X | 2004-03-17 | ||
| CN200410018757 | 2004-03-17 | ||
| CNB2005100552648A CN100442049C (en) | 2004-03-17 | 2005-03-17 | Method for measuring fingerprint pattern of compound salvia dropping pills |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1670525A true CN1670525A (en) | 2005-09-21 |
| CN100442049C CN100442049C (en) | 2008-12-10 |
Family
ID=35041861
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100552648A Active CN100442049C (en) | 2004-03-17 | 2005-03-17 | Method for measuring fingerprint pattern of compound salvia dropping pills |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100442049C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110274978A (en) * | 2018-03-13 | 2019-09-24 | 天士力医药集团股份有限公司 | The more saponin constituent content assaying methods of Radix Notoginseng in a kind of QISHEN YIQI DIWAN |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| HUP0303906A3 (en) * | 2000-12-22 | 2005-05-30 | Cardionat Inc Whitestone | Herbal composition for angina pectoris, method to prepare same and uses thereof |
| CN100381813C (en) * | 2004-03-16 | 2008-04-16 | 天津天士力制药股份有限公司 | Quality control of compound Danshen root drops |
-
2005
- 2005-03-17 CN CNB2005100552648A patent/CN100442049C/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110274978A (en) * | 2018-03-13 | 2019-09-24 | 天士力医药集团股份有限公司 | The more saponin constituent content assaying methods of Radix Notoginseng in a kind of QISHEN YIQI DIWAN |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100442049C (en) | 2008-12-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1895617A (en) | Kidney-warming and heart-nourishing Chinese-medicinal preparation, its making method and quality control | |
| CN1669573A (en) | Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases and preparing process thereof | |
| CN101036748A (en) | Quality control method of the Yixinshu Chinese traditional medicine for supplementing qi and for promoting blood circulation by removing blood stasis | |
| CN1670529A (en) | Method for constructing Compound Xueshuantong preparation HPLC fingerprint pattern and method standard fingerprint pattern thereof | |
| CN1806846A (en) | Chinese medicinal composition, its preparation process and quality control method | |
| CN1785380A (en) | Quality control method of Chinese medicinal preparation | |
| CN1843425A (en) | Dispersion tablet for getting brain and heart unobstructed and preparation method and its quality control method | |
| CN1679870A (en) | Quality control of compound Danshen root drops | |
| CN1876089A (en) | A pharmaceutical composition for treating kidney-qi deficiency syndrome and preparation method thereof | |
| CN102068657B (en) | Quality control method of venation relaxing pill as Chinese herbal preparation | |
| CN1670524A (en) | Method for measuring fingerprint pattern of compound salvia dropping pills | |
| CN1843424A (en) | Cassia twig tuckahoe effervescence tablet and preparation method and its quality control method | |
| CN1679811A (en) | Danshen root extract, production, medicine preparation and inspection thereof | |
| CN1872199A (en) | Composition of Chinese traditional medicine, and preparation method | |
| CN1785347A (en) | Quality control method of Chinese medicinal preparation for treating child hyperpyrexia | |
| CN1670525A (en) | Method for measuring fingerprint pattern of compound salvia dropping pills | |
| CN1679871A (en) | Quality control of compound Danshen root drops | |
| CN1785220A (en) | Method for quality control of blood-bonifying Chinese angelica prepn. | |
| CN1785281A (en) | Quality control method of spirit quieting oral liquid preparation | |
| CN1257710C (en) | Preparation made from effective position of red sage root, and its prepn. method | |
| CN1472532A (en) | Method for determining salviolic acid beta content in red sage root | |
| CN1562310A (en) | Soft capsule preparation of Tibet medicine 'unique taste' | |
| CN1586612A (en) | Process for preparing granular powder for treating blood stasis disease and quality control method | |
| CN101066437A (en) | Quality control method for compound cantharis oral liquid | |
| CN1857588A (en) | Quality control method for Xianlinggubao preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: TASLY PHARMACEUTICAL GROUP CO., LTD. Free format text: FORMER NAME: TIANJIN TASLY PHARMACEUTICAL CO., LTD. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee after: Tasly Pharmaceutical Group Co., Ltd. Address before: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee before: Tianjin Tianshili Pharmaceutical Co., Ltd. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee after: Tasly Pharmaceutical Group Limited by Share Ltd Address before: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee before: Tasly Pharmaceutical Group Co., Ltd. |
|
| CP01 | Change in the name or title of a patent holder |