CN104569199B - A kind of assay method of Radix Astragali finger printing - Google Patents
A kind of assay method of Radix Astragali finger printing Download PDFInfo
- Publication number
- CN104569199B CN104569199B CN201410857192.8A CN201410857192A CN104569199B CN 104569199 B CN104569199 B CN 104569199B CN 201410857192 A CN201410857192 A CN 201410857192A CN 104569199 B CN104569199 B CN 104569199B
- Authority
- CN
- China
- Prior art keywords
- radix astragali
- peak
- reference substance
- finger printing
- chromatographic
- 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.)
- Active
Links
- 239000009636 Huang Qi Substances 0.000 title claims abstract description 112
- 238000007639 printing Methods 0.000 title claims abstract description 42
- 238000003556 assay Methods 0.000 title claims abstract description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 114
- 238000012360 testing method Methods 0.000 claims abstract description 47
- 239000013558 reference substance Substances 0.000 claims abstract description 40
- 238000000605 extraction Methods 0.000 claims abstract description 23
- 239000000706 filtrate Substances 0.000 claims abstract description 20
- 239000000843 powder Substances 0.000 claims abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 10
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 239000012982 microporous membrane Substances 0.000 claims abstract description 8
- ZZAJQOPSWWVMBI-UHFFFAOYSA-N Calycosin Natural products C1=C(O)C(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZZAJQOPSWWVMBI-UHFFFAOYSA-N 0.000 claims description 26
- HKQYGTCOTHHOMP-UHFFFAOYSA-N formononetin Chemical compound C1=CC(OC)=CC=C1C1=COC2=CC(O)=CC=C2C1=O HKQYGTCOTHHOMP-UHFFFAOYSA-N 0.000 claims description 21
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 claims description 16
- 230000014759 maintenance of location Effects 0.000 claims description 13
- 229930182490 saponin Natural products 0.000 claims description 13
- 150000007949 saponins Chemical class 0.000 claims description 13
- SMDOOINVMJSDPS-UHFFFAOYSA-N Astragaloside Natural products C1=C(O)C(OC)=CC(C2=C(C(=O)C3=C(O)C=C(O)C=C3O2)OC2C(C(OC3C(C(O)C(O)C(CO)O3)O)C(O)C(CO)O2)O)=C1 SMDOOINVMJSDPS-UHFFFAOYSA-N 0.000 claims description 12
- QMNWISYXSJWHRY-XWJCTJPOSA-N astragaloside 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)C4[C@@H](O[C@H]4[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O4)O)CC3[C@]2(C)C[C@@H]1O QMNWISYXSJWHRY-XWJCTJPOSA-N 0.000 claims description 12
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 12
- LAWPHHZXTUPSDG-UHFFFAOYSA-N Astragaloside I Natural products CC(=O)OC1C(O)COC(OC2CCC34CC35CCC6(C)C(C)(CCC6(O)C7(C)CCC(O7)C(C)(C)O)C5CC(OC8OC(CO)C(O)C(O)C8O)C4C2(C)C)C1OC(=O)C LAWPHHZXTUPSDG-UHFFFAOYSA-N 0.000 claims description 11
- KXHCYYSIAXMSPA-UHFFFAOYSA-N Astragaloside-I Chemical compound CC(=O)OC1C(OC(=O)C)C(O)COC1OC1C(C)(C)C2C(OC3C(C(O)C(O)C(CO)O3)O)CC3C4(C)CC(O)C(C5(C)OC(CC5)C(C)(C)O)C4(C)CCC43CC42CC1 KXHCYYSIAXMSPA-UHFFFAOYSA-N 0.000 claims description 11
- MGJLSBDCWOSMHL-WFMNFSIZSA-N Ononin Natural products O(C)c1ccc(C=2C(=O)c3c(OC=2)cc(O[C@H]2[C@@H](O)[C@@H](O)[C@H](O)[C@H](CO)O2)cc3)cc1 MGJLSBDCWOSMHL-WFMNFSIZSA-N 0.000 claims description 11
- HJCCJZGOBHFQSX-UHFFFAOYSA-N cyclosieversioside B Natural products CC(CCC(O)C(C)(C)OC1OC(CO)C(O)C(O)C1O)C2C(CC3(C)C4CC(O)C5C(C)(C)C(CCC56CC46CCC23C)OC7OCC(O)C(O)C7O)OC8OC(CO)C(O)C(O)C8O HJCCJZGOBHFQSX-UHFFFAOYSA-N 0.000 claims description 11
- 238000001514 detection method Methods 0.000 claims description 11
- RIKPNWPEMPODJD-UHFFFAOYSA-N formononetin Natural products C1=CC(OC)=CC=C1C1=COC2=CC=CC=C2C1=O RIKPNWPEMPODJD-UHFFFAOYSA-N 0.000 claims description 11
- MGJLSBDCWOSMHL-MIUGBVLSSA-N ononin Chemical compound C1=CC(OC)=CC=C1C1=COC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=CC=C2C1=O MGJLSBDCWOSMHL-MIUGBVLSSA-N 0.000 claims description 11
- MGJLSBDCWOSMHL-UHFFFAOYSA-N ononoside Natural products C1=CC(OC)=CC=C1C1=COC2=CC(OC3C(C(O)C(O)C(CO)O3)O)=CC=C2C1=O MGJLSBDCWOSMHL-UHFFFAOYSA-N 0.000 claims description 11
- 229930182478 glucoside Natural products 0.000 claims description 8
- -1 calycosin glucoside Chemical class 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 5
- WACBUPFEGWUGPB-MIUGBVLSSA-N calycosin-7-O-beta-D-glucoside Chemical compound C1=C(O)C(OC)=CC=C1C1=COC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)=CC=C2C1=O WACBUPFEGWUGPB-MIUGBVLSSA-N 0.000 claims description 3
- WACBUPFEGWUGPB-UHFFFAOYSA-N calycosin-7-O-beta-D-glucoside Natural products C1=C(O)C(OC)=CC=C1C1=COC2=CC(OC3C(C(O)C(O)C(CO)O3)O)=CC=C2C1=O WACBUPFEGWUGPB-UHFFFAOYSA-N 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims 1
- 235000009392 Vitis Nutrition 0.000 claims 1
- 241000219095 Vitis Species 0.000 claims 1
- 150000002576 ketones Chemical class 0.000 claims 1
- RYVMUASDIZQXAA-UHFFFAOYSA-N pyranoside Natural products O1C2(OCC(C)C(OC3C(C(O)C(O)C(CO)O3)O)C2)C(C)C(C2(CCC3C4(C)CC5O)C)C1CC2C3CC=C4CC5OC(C(C1O)O)OC(CO)C1OC(C1OC2C(C(OC3C(C(O)C(O)C(CO)O3)O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OCC(O)C(O)C1O RYVMUASDIZQXAA-UHFFFAOYSA-N 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 16
- 239000000203 mixture Substances 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 9
- 239000000047 product Substances 0.000 abstract description 6
- 238000003908 quality control method Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 45
- 239000012071 phase Substances 0.000 description 17
- 239000002904 solvent Substances 0.000 description 14
- 239000000523 sample Substances 0.000 description 13
- 238000010521 absorption reaction Methods 0.000 description 12
- 238000002360 preparation method Methods 0.000 description 12
- 238000005070 sampling Methods 0.000 description 12
- 238000004128 high performance liquid chromatography Methods 0.000 description 10
- 235000017709 saponins Nutrition 0.000 description 10
- 239000012467 final product Substances 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 8
- 238000001228 spectrum Methods 0.000 description 8
- 239000000284 extract Substances 0.000 description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 6
- 230000003595 spectral effect Effects 0.000 description 6
- 239000008346 aqueous phase Substances 0.000 description 5
- 229960004756 ethanol Drugs 0.000 description 5
- 241000545442 Radix Species 0.000 description 4
- 229960000935 dehydrated alcohol Drugs 0.000 description 4
- 229930182470 glycoside Natural products 0.000 description 4
- 150000002338 glycosides Chemical class 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000003672 processing method Methods 0.000 description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 3
- 241000045403 Astragalus propinquus Species 0.000 description 3
- 206010019133 Hangover Diseases 0.000 description 3
- 241000700159 Rattus Species 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 229930003944 flavone Natural products 0.000 description 3
- 235000011949 flavones Nutrition 0.000 description 3
- 235000019253 formic acid Nutrition 0.000 description 3
- 239000012567 medical material Substances 0.000 description 3
- 239000000401 methanolic extract Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000013112 stability test Methods 0.000 description 3
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 2
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 description 2
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 235000006533 astragalus Nutrition 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 150000002212 flavone derivatives Chemical class 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 238000005464 sample preparation method Methods 0.000 description 2
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical compound C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 description 2
- 235000021286 stilbenes Nutrition 0.000 description 2
- 230000009182 swimming Effects 0.000 description 2
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 241000255789 Bombyx mori Species 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 206010027514 Metrorrhagia Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000596467 Ononis Species 0.000 description 1
- DCTLJGWMHPGCOS-UHFFFAOYSA-N Osajin Chemical compound C1=2C=CC(C)(C)OC=2C(CC=C(C)C)=C(O)C(C2=O)=C1OC=C2C1=CC=C(O)C=C1 DCTLJGWMHPGCOS-UHFFFAOYSA-N 0.000 description 1
- 230000010718 Oxidation Activity Effects 0.000 description 1
- 241000219506 Phytolacca Species 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- OUZCFMSJGDEXRT-UHFFFAOYSA-N Scandinone Natural products O=C1C=2C(OC)=C(CC=C(C)C)C=3OC(C)(C)C=CC=3C=2OC=C1C1=CC=C(O)C=C1 OUZCFMSJGDEXRT-UHFFFAOYSA-N 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 208000022531 anorexia Diseases 0.000 description 1
- 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 1
- 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 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012846 chemical reference substance Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 208000019902 chronic diarrheal disease Diseases 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 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 1
- 230000007423 decrease Effects 0.000 description 1
- 206010061428 decreased appetite Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 210000001840 diploid cell Anatomy 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002118 epoxides Chemical class 0.000 description 1
- QOLIPNRNLBQTAU-UHFFFAOYSA-N flavan Chemical compound C1CC2=CC=CC=C2OC1C1=CC=CC=C1 QOLIPNRNLBQTAU-UHFFFAOYSA-N 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- NNQSGBRGJHSRFN-UHFFFAOYSA-N isoflavan Chemical compound C1OC2=CC=CC=C2CC1C1=CC=CC=C1 NNQSGBRGJHSRFN-UHFFFAOYSA-N 0.000 description 1
- 235000002324 isoflavanes Nutrition 0.000 description 1
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 1
- 235000008696 isoflavones Nutrition 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003387 muscular Effects 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000011524 similarity measure Methods 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 150000008505 β-D-glucopyranosides Chemical class 0.000 description 1
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The present invention relates to the assay method of a kind of Radix Astragali finger printing, including: (1) takes powder in the Radix Astragali, accurately weighed, adds methanol, supersound extraction 60min, filters, and filtrate volatilizes, residue add methanol dissolve, constant volume with filtering with microporous membrane, take subsequent filtrate, obtain need testing solution；(2) reference substance solution is prepared；(3) precision draws need testing solution and reference substance solution respectively, injects high performance liquid chromatograph and measures, obtains Radix Astragali finger printing.The peak of gained Radix Astragali finger printing of the present invention is many, and peak shape is good, peak good separating effect, it is easy to differentiate, similarity is high, accurately and reliably, determines that 26 chromatographic peaks, for total chromatographic peak, determine 7 compositions through control comparisons product solution chromatographic peak；The inventive method is easy, quick, accurate, stable, reliable, can be used for the quality control of Radix Astragali sample, has a good application prospect.
Description
Technical field
The invention belongs to Chinese medicine fingerprint field, particularly to the assay method of a kind of Radix Astragali finger printing.
Background technology
The Original plant of the Radix Astragali is leguminous plant Radix Astagali Astragalus membranaceus (Fisch.)
Bge.var.mongholicus (Bge.) hsiao or the root of Radix Astragali A.membranaceus (Fisch.) Bge..In the Radix Astragali
Containing multiple chemical composition, such as compositions such as flavone, saponins and polysaccharides. flavones ingredient is main such as awns with osajin composition
Handle xanthin, 3'-hydroxyl formononetin (calycosin) and glycoside, 2', 3'-dihydroxy-7,4'-dimethoxy
Isoflavone, 7,2'-dihydroxy-3', 4'-dimethoxy different Radix Astragali flavane and glycoside, 7,3'-dihydroxy-4', 5'-bis-
Methoxyl group isoflavan, 3-hydroxyl-9,10-dimethoxy Lignum pterocarpi indici alkane and glucoside thereof etc. stronger the resisting of some of which composition tool
Oxidation activity. saponin component has astragaloside I～VIII and Sayasaponin Ⅰ；Astragaloside (i.e. astragaloside Ⅳ) and Radix Astragali second
Glycosides is main component therein.
The Radix Astragali has invigorating QI to consolidate the body surface resistance, diuresis poison holding, evacuation of pus, effect of expelling pus and promoting granulation.Weak for the deficiency of vital energy, anorexia and loose stool,
Sinking of QI of middle-JIAO, chronic diarrhea proctoptosis, metrorrhagia of having blood in stool, exterior deficiency spontaneous perspiration, carbuncle difficulty is burst, and bursts for a long time and does not holds back, and blood deficiency and yellow complexion, interior-heat is quenched one's thirst.Modern
Research shows that the Radix Astragali has enhancing non-specific immunity function, and the Radix Astragali can dramatically increase the total white blood cells in blood, in promotion
Property granulocyte and the phagocytic function of macrophage and sterilizing ability.The enhancing specific immune function Radix Astragali can be remarkably reinforced cell and exempt from
Epidemic disease, promotes the lymphocyte transformation that PHA, COnA, PWM (phytolacca american) cause.Strengthening the effect of resisting fatigue, the Radix Astragali can strengthen greatly
Mus and the muscular strength of mice.Rat oral gavage Astragalus membranaceus 6 days or 14 days, can strengthen the fatigue proof effect of rats'swimming, and make trip
Swimming stress rats plasma cortisol substantially increases, and exceedes blank stress group and the level of Normal group.Slow down nature to decline
Old effect, the Radix Astragali can extend the average life of silkworm and fruit bat, slows down naturally declining of HFL's diploid cell In vitro culture
Old process, makes cell life extension reach for 98 generations, and matched group is only 61-66 generation, makes life l/3.
Summary of the invention
The technical problem to be solved is to provide the assay method of a kind of Radix Astragali finger printing, and the method gained is yellow
The peak of stilbene finger printing is many, and peak shape is good, peak good separating effect, it is easy to differentiate, similarity is high, accurately and reliably, determines 26 chromatographs
Peak is total chromatographic peak, determines 7 compositions through control comparisons product solution chromatographic peak.
A kind of assay method of the Radix Astragali finger printing of the present invention, including:
(1) powder in the Radix Astragali is taken, accurately weighed, add methanol, 35KHz supersound extraction 60min, filter, filtrate volatilizes, residue
Add methanol dissolve, constant volume with filtering with microporous membrane, take subsequent filtrate, obtain need testing solution；
(2) calycosin glucoside reference substance, ononin reference substance, 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-are taken
β-D glucopyranoside reference substance, calycosin reference substance, astragaloside reference substance, Radix Astragali saponin III reference substance, Radix Ononis hircinae
Element reference substance, astragaloside I reference substance, accurately weighed, it is separately added into methanol and makes reference substance solution；
(3) precision draws need testing solution and reference substance solution respectively, injects high performance liquid chromatograph and measures, obtains the Radix Astragali
Finger printing；Wherein, chromatographic condition is: chromatographic column: Athena C18-WP chromatographic column；Column temperature: 30 DEG C；Detection wavelength: 208nm；
Sampling volume: 10 μ l；Flow velocity: 1ml min-1；Flowing phase: mobile phase A is water, and Mobile phase B is acetonitrile；Gradient elution program: 0
～20min:5%B → 20%B, 20～70min:20%B → 40%B, 70～85min:40%B → 60%B.
In the Radix Astragali in described step (1), powder is 1g:20ml with the ratio of methanol.
The aperture of the microporous filter membrane in described step (1) is 0.22 μm.
Reference substance in described step (2) is 0.05～1.5mg:1ml with the ratio of methanol.
Reference substance solution in described step (2) is particularly as follows: every 1ml is respectively containing Calycosin-7-O-BETA-D-glucoside 0.2mg, ononin
0.05mg, 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-β-D glucopyranoside 0.08mg, calycosin 0.2mg, astragaloside
1.4mg, Radix Astragali saponin III 1.2mg, formononetin 0.05mg, the mixed solution of astragaloside I 1.2mg.
In Radix Astragali finger printing in described step (3), 26 chromatographic peaks are confirmed as total chromatographic peak, by with compare
The comparison of product retention time, determines that peak 10 is for calycosin glucoside；Peak 12 is ononin；Peak 13 is 9,10-diformazan
Epoxide Lignum pterocarpi indici alkane-3-O-β-D glucopyranoside；Peak 16 is calycosin；Peak 20 is astragaloside；Peak 21 is Radix Astragali saponin
Ⅲ；Peak 22 is formononetin；Peak 26 is astragaloside I.
Beneficial effect
The peak of gained Radix Astragali finger printing of the present invention is many, and peak shape is good, peak good separating effect, it is easy to differentiate, similarity is high, accurate
The most reliable, determine that 26 chromatographic peaks, for total chromatographic peak, determine 7 compositions through control comparisons product solution chromatographic peak；This
Bright method is easy, quick, accurate, stable, reliable, can be used for the quality control of Radix Astragali sample, has a good application prospect.
Accompanying drawing explanation
Fig. 1 is 7 kinds of composition reference substances full wavelength scanner result and absorption maximum in ultraviolet interval present in the Radix Astragali
Wavelength；Wherein, a is calycosin glucoside, and b is ononin, and c is calycosin, and d is astragaloside, and e is the Radix Astragali
Saponin III, f is formononetin, and g is astragaloside I；
Fig. 2 is Radix Astragali finger printing under the conditions of different wave length；
Fig. 3 is the Radix Astragali fingerprint chromatogram of Kromasil 100-5C18 chromatographic column；Wherein, a-c is different gradient
Program；
Fig. 4 is the Radix Astragali fingerprint chromatogram of Athena C18 chromatographic column；Wherein, a-b is different gradient program；
Fig. 5 is the Radix Astragali fingerprint chromatogram of Waters XSelect CSH C18 chromatographic column；
Fig. 6 is Athena C18-WP chromatographic column Radix Astragali fingerprint chromatogram；Wherein, a-c is different gradient program；
Fig. 7 is Radix Astragali HPLC chromatogram under different column temperature；
Fig. 8 is different extracting mode need testing solution HPLC chromatogram；
Fig. 9 is different solvents need testing solution HPLC chromatogram；
Figure 10 is various extracting conditions need testing solution HPLC chromatogram；
Figure 11 is variable concentrations need testing solution, different sampling volume HPLC chromatogram；
Figure 12 is mixed chemical reference substance finger printing；
Figure 13 is different sources batch Radix Astragali finger printing；
Figure 14 is Radix Astragali standard finger-print；
Figure 15 is that two batch Radixs Astragali compare with Radix Astagali and Radix Astragali finger printing.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate the present invention
Rather than restriction the scope of the present invention.In addition, it is to be understood that after having read the content that the present invention lectures, people in the art
The present invention can be made various changes or modifications by member, and these equivalent form of values fall within the application appended claims equally and limited
Scope.
Embodiment 1
1. the selection of instrument parameter
(1) selection of wavelength is detected
To 7 kinds of composition reference substances full wavelength scanner result and maximum absorption wavelengths in ultraviolet interval present in the Radix Astragali
As shown in Fig. 1 a-g.It can be seen that calycosin glucoside maximum absorption wave a length of 220nm, 258nm, ononin and
The a length of 250nm of formononetin maximum absorption wave, calycosin maximum absorption wave a length of 220nm, 258nm, astragaloside, the Radix Astragali
Saponin III and astragaloside I end absorption maximum intensity.So, the ultraviolet full wavelength absorption scanning of each reference substance of Integrated comparative
Figure, Radix Astragali finger printing detection wavelength should select at end absorption, investigates the prepared sample of the Radix Astragali below respectively at five
The finger printing obtained is detected to selecting optimum absorb wavelength under wavelength (203nm, 208nm, 265nm, 250nm, 280nm).
Sample preparation methods: take powder in the 2.0g Radix Astragali, accurately weighed, add methanol 40ml ultrasonic (53KHz, 280W) and extract
60min, filters, and filtrate volatilizes, and residue proper amount of methanol is dissolved and is settled to 10ml.
Chromatographic condition: volume flow: 1.0ml min-1；Sampling volume: 10 μ l；Column temperature: 30 DEG C；Flowing phase: water (A)-second
Nitrile (B), gradient elution: 0～5min:5%B, 5～12min:5%B → 10%B, 12～60min:10%B → 30%B, 60～
90min:30%B → 55%B, 90～95min:55%B → 100%B.Finger printing result is as shown in Figure 2.It can be seen that
Under 208nm, the finger printing peak height of detection is higher, and baseline drift is less, therefore selects 208nm as the inspection of Radix Astragali finger printing
Survey wavelength.(2) chromatographic column selection and flowing phase elution program optimization
1. Kromasil 100-5C18 chromatographic column
Selecting appropriate solvent, strive for that composition as much as possible is detected, this experiment initial option methanol is molten as extracting
Agent, carries out supersound extraction, and carries out efficient liquid phase chromatographic analysis powder in medical material.
The preparation of need testing solution: take powder 2.0g in the Radix Astragali, accurately weighed, add methanol 30ml ultrasonic (53KHz, 280W) and carry
Take 60min, let cool, supply the weight of solvent of less loss, cross 0.22 μm microporous filter membrane, take subsequent filtrate, to obtain final product.
Chromatographic condition: column temperature is 30 DEG C, flowing is water (A)-acetonitrile (B) mutually, and flow velocity is 0.8ml min-1, sampling volume
It is 10 μ l, gradient elution program: 0～70min:5%B → 100%B.Detection wavelength is 190-400nm full wavelength scanner.Record
Shown in HPLC chromatogram as Fig. 3 a.It can be seen that after 40min (B phase reaches about 55%), only 1 characteristic peak occurs.
Continue to optimize chromatographic condition.
It is as follows that above-mentioned liquid-phase condition only changes gradient program: 0～10min:5%B, 10～15min:5%B → 15%
B, 15～35min:15%B → 35%B, 35～50min:30%B → 60%B, 50～65min:60%B → 100%B.Gained
Chromatogram is as shown in Figure 3 b.It can be seen that the drift of 208nm wave spectrum is serious, 220nm spectrogram has 13 peaks, and 254nm spectrogram has 10
, wherein there are two peak hangovers at peak, prolong before a peak.Still need to further eluent gradient change is adjusted, so that chromatographic peak
Preferably separated.
In process of the test, utilize DAD detector 3D spectrogram that sample is compared from the chromatographic absorption of 190nm-400nm
Relatively, result shows, the methanol extract liquid of the Radix Astragali has obvious end absorption, and at 208nm, the absorption response value of each composition is higher, has
Several peaks prolong or conditions of streaking before existing, it is considered to aqueous phase changes 0.2% formic acid into.
Flowing phase: 0.2% formic acid (A)-acetonitrile (B)；Gradient program: 0～5min:5%B, 5～35min:5%B →
35%B, 35～55min:35%B → 60%B, 55～70min:60%B → 100%B.Gained spectrogram is as shown in Figure 3 c.Permissible
Finding out, after adding formic acid in aqueous phase, the drift of 240nm, 203nm, 208nm wave spectrum is serious, and 254nm, 260nm, 280nm wave spectrum is more flat
Surely.If optimizing further, drift serious problems still cannot solve, therefore aqueous phase should select pure water.
2. Athena C18 chromatographic column
Flowing phase: water (A)-acetonitrile (B)；Gradient program is: 0～5min:5%B, 5～40min:5%B → 35%
B, 40～55min:35%B → 60%B, 55～70min:60%B → 100%B.Remaining chromatographic condition is ibid；Test sample processes
Method is ibid.Gained spectrogram as shown in fig. 4 a it can be seen that under these conditions wave spectrum relatively steady, drift about little, after 60min, have valency
Value peak is few.Continue to optimize gradient program.
Gradient program is: 0～8min:5%B, 8～12min:5%B → 12%B, 12～55min:12%B →
40%B, 55～75min:40%B → 55%B, 75～85min:55%B → 65%B.Spectrogram is as shown in Figure 4 b.It can be seen that
Having 19 peaks under these conditions, there is hangover, front phenomenon of prolonging in indivedual peaks, without peak after 60min.
3. Waters XSelect CSH C18 chromatographic column
Flowing phase: water (A)-acetonitrile (B)；Gradient program is: 0～10min:5%B → 15%B, 10～50min:
15%B → 35%B, 50～75min:35%B → 60%B.Remaining chromatographic condition is ibid；Test sample processing method is ibid.Gained
Spectrogram is as shown in Figure 5.It can be seen that this chromatographic column is for going out the number at peak, separating degree etc. without much improvement.
4. Athena C18-WP chromatographic column
The preparation of need testing solution: take powder 2.0g in the Radix Astragali, accurately weighed, add methanol 40ml ultrasonic (53KHz, 280W) and carry
Taking 30min, filter, filtrate volatilizes, and residue methanol dissolves and be settled to 5ml, crosses 0.22 μm microporous filter membrane, takes subsequent filtrate, i.e.
?.
Flowing phase: water (A)-acetonitrile (B)；Gradient program is: 0～5min:0%B, 5～90min:0%B → 60%
B.Remaining chromatographic condition is ibid；Test sample processing method is ibid.Remaining chromatographic condition is ibid.Gained spectrogram is as shown in Figure 6 a.Can
To find out, going out peak more under this chromatographic condition, peak shape is preferable, without substantially hangover, leading peak, therefore selects this chromatographic column for analyzing
The finger printing of the Radix Astragali.But using 100% aqueous phase as initial flow phase time, miscellaneous peak height in said method, and divide with postpeak
From without much improvement, extend analysis time simultaneously, therefore examination changes work 95% aqueous phase as initial flow phase.
Flowing phase: water (A)-acetonitrile (B)；Gradient program is: 0～8min:5%B, 8～10min:5%B → 12%
B, 10～35min:12%B → 20%B, 35～65min:20%B → 40%B, 65～75min:40%B → 50%B.Remaining color
Spectral condition is ibid；Test sample processing method is ibid.Spectrogram is as shown in Figure 6 b.It can be seen that occur without peak in 8-15min, 15-
In 18min, peak is intensive.Continue to optimize gradient.
Gradient program is: 0～20min:5%B → 20%B, 20～70min:20%B → 40%B, 70～85min:
40%B → 60%B.Remaining chromatographic condition is ibid；Test sample processing method is ibid.Spectrogram is as fig. 6 c.Therefore select Athena
C18-WP chromatographic column, the spectrogram effect obtained by the gradient determined is preferable.
(3) selection of chromatographic column temperature
The preparation of need testing solution: take powder 2.0g in the Radix Astragali, accurately weighed, add methanol 40ml ultrasonic (53KHz, 280W)
Extracting 30min, extracting solution filters, and filtrate volatilizes, and residue adds methanol and dissolves and be settled to 10ml, by 0.22 μm microporous filter membrane mistake
Filter, to obtain final product.
Chromatographic condition: chromatographic column is Athena C18-WP chromatographic column, flow velocity is 1ml min-1, sampling volume is 10 μ l,
Detection wavelength is 208nm, and flow phase: water (A)-acetonitrile (B)；Gradient program is: 0～20min:5%B → 20%B, 20～
70min:20%B → 40%B, 70～85min:40%B → 60%B.Different need testing solutions are respectively at 25 DEG C, 30 DEG C, 35 DEG C
Detecting under three column temperatures, finger printing is as shown in Figure 7.Understanding, when column temperature selects 25 DEG C or 35 DEG C, characteristic peak quantity is the most relatively
Few, therefore column temperature selects 30 DEG C preferably.
2. the selection and optimization of need testing solution preparation condition
(1) selection of extracting mode
Two kinds of need testing solutions that supersound extraction and reflux, extract, are obtained by the present embodiment carry out efficient liquid phase chromatographic analysis,
By the finger printing of relatively different extracting modes, to determine applicable extracting method.
Test sample preparation method: take 2 parts of each 2.0g of powder in the Radix Astragali, accurately weighed, it is separately added into methanol 40ml, respectively according to table
A kind of method in 1 is extracted, and extracting solution filters and volatilizes, and residue methanol dissolves and is settled to 10ml, micro-by 0.22 μm
Hole membrane filtration, takes subsequent filtrate, to obtain final product.
Table 1 need testing solution extracting method table
| Numbering | Extracting method | Extraction time |
| 1 | Backflow | 180min |
| 2 | Ultrasonic | 60min |
Chromatographic condition: chromatographic column is Athena C18-WP chromatographic column, column temperature is 30 DEG C, and flowing is water (A)-acetonitrile mutually
(B), flow velocity is 1ml min-1, sampling volume is 10 μ l, and detection wavelength is 208nm, gradient program: 0～5min:5%B,
5～12min:5%B → 10%B, 12～60min:10%B → 30%B, 60～90min:30%B → 55%B, 90～95min:
55%B → 100%B.Result is shown in Fig. 8.It can be seen that in the quantity of shown chromatographic peak, two kinds of extracting method zero differences；?
In the intensity of chromatographic peak, circumfluence method is slightly higher, but circumfluence method operation is cumbersome, and therefore, the present embodiment selects ultrasonic as extraction side
Formula.
(2) selection of Extraction solvent
This experiment selects with the solvent of 6 kinds of variable concentrations: dehydrated alcohol, 50% ethanol, 70% ethanol, 50% methanol,
70% methanol and methanol, respectively as Extraction solvent, carry out supersound extraction, and carry out efficient liquid phase chromatographic analysis powder in medical material.
In chromatogram, the number of chromatographic peak, peak shape, separating degree, absorption intensity, baseline characteristic are as inspection target, extract different solvents
The chromatogram of liquid is analyzed comparing.By the finger printing of relatively different need testing solutions, filter out the extraction being more suitable for
Solvent.
The preparation of need testing solution: take 6 parts of each 2.0g of powder in the Radix Astragali, accurately weighed, it is separately added into Extraction solvent (being shown in Table 2)
40ml, ultrasonic (53KHz, 280W) extracts 30min, and extracting solution filters, and filtrate volatilizes, and residue is with the solvent identical with Extraction solvent
Carry out dissolving and being settled to 5ml, with 0.22 μm filtering with microporous membrane, take subsequent filtrate, to obtain final product.
The Extraction solvent table that table 2 need testing solution selects
| Numbering | Extraction solvent |
| 1 | Dehydrated alcohol |
| 2 | 50% ethanol |
| 3 | 70% ethanol |
| 4 | 50% methanol |
| 5 | 70% methanol |
| 6 | Methanol |
Chromatographic condition: chromatographic column is Athena C18-WP chromatographic column, column temperature is 30 DEG C, and flowing is water (A)-acetonitrile mutually
(B), flow velocity is 1ml min-1, sampling volume is 10 μ l, and detection wavelength is 208nm, gradient program: 0～5min:5%B,
5～12min:5%B → 10%B, 12～60min:10%B → 30%B, 60～90min:30%B → 55%B, 90～95min:
55%B → 100%B.The finger printing of need testing solution 1-solution 6 is as shown in Figure 9.It can be seen that 50% ethanol, 70% second
Alcohol, 50% methanol, 70% methanol extraction rate are relatively low, and wave spectrum baseline drift is serious.The wave spectrum base of dehydrated alcohol and methanol extract liquid
Line is the most stable, but the chromatographic peak of dehydrated alcohol extraction liquid is less, and the separating degree of chromatographic peak is poor, and the chromatographic peak of methanol extract liquid
Intensity is higher, and the separating degree of chromatographic peak and peak shape more preferable.Therefore, initial option methanol is as Extraction solvent.
(3) selection of extraction conditions (extraction time and supersonic frequency)
The preparation of need testing solution: take 4 parts of each 2.0g of powder in the Radix Astragali, accurately weighed, it is separately added into methanol 40ml, by table 3
Shown in supersound extraction different time at different frequencies, extracting solution filters and volatilizes, and residue methanol dissolves, is settled to 5ml also
With 0.22 μm filtering with microporous membrane, take subsequent filtrate, to obtain final product.
Table 3 need testing solution ultrasonic time, frequency meter
Chromatographic condition: chromatographic column is Athena C18-WP chromatographic column, column temperature is 30 DEG C, and flowing is water (A)-acetonitrile mutually
(B), flow velocity is 1ml min-1, sampling volume is 10 μ l, and detection wavelength is 208nm, and flow phase: water (A)-acetonitrile (B)；Eluting
Gradient program is: 0～5min:5%B, 5～12min:5%B → 10%B, 12～60min:10%B → 30%B, 60～
90min:30%B → 55%B, 90～95min:55%B → 100%B.Finger printing such as Figure 10 institute of different need testing solutions
Show.It can be seen that 90min with 60min extraction ratio is close, being all higher than 30min extraction ratio, 35KHz extraction ratio is more than 53KHz, therefore
Consider selection ultrasonic time and frequency is respectively 35KHz, 60min.
(4) need testing solution concentration and the selection of sampling volume
The preparation of need testing solution: take 3 parts of powder (being shown in Table 4) in the Radix Astragali, accurately weighed, it is separately added into methanol 40ml, ultrasonic
(35KHz, 280W) extracts 60min, and extracting solution filters, and filtrate volatilizes, and residue methanol dissolves and is settled to 10ml, uses 0.22 μm
Filtering with microporous membrane, takes subsequent filtrate, to obtain final product.
Opaque amount, sampling volume table in table 4 Radix Astragali
Chromatographic condition: chromatographic column is Athena C18-WP chromatographic column, column temperature is 30 DEG C, and flowing is water (A)-acetonitrile mutually
(B), flow velocity is 1ml min-1, sampling volume is shown in Table 1-10, and detection wavelength is 208nm, and flow phase: water (A)-acetonitrile (B)；Wash
De-Gradient program is: 0～20min:5%B → 20%B, 20～70min:20%B → 40%B, 70～85min:40%B →
60%B.The finger printing of different need testing solutions is as shown in figure 11.Known by figure: take powder in the 2.0g Radix Astragali and prepare test sample in accordance with the law
Solution, takes 10 μ l sample introduction effects best.
3. brief summary
Through above-mentioned experiment, determine that Radix Astragali fingerprint map analyzing condition and need testing solution preparation method are as follows:
Radix Astragali HPLC fingerprint map analyzing condition: chromatographic column: Athena C18-WP chromatographic column；Column temperature: 30 DEG C；Detection ripple
Long: 208nm；Sampling volume: 10 μ l；Flow velocity is 1ml min-1；Flowing phase: water (A)-acetonitrile (B)；Gradient elution program: 0～
20min:5%B → 20%B, 20～70min:20%B → 40%B, 70～85min:40%B → 60%B.
The preparation of need testing solution: take powder 2.0g in the Radix Astragali, accurately weighed, add methanol 40ml ultrasonic (35KHz, 280W)
Extracting 60min, extracting solution filters, and filtrate volatilizes, and residue methanol dissolves and is settled to 10ml, with 0.22 μm filtering with microporous membrane
Extracting solution, to obtain final product.
Embodiment 2
1. the preparation of solution
(1) preparation of need testing solution: take each 2.0g of powder in the different batches Radix Astragali, accurately weighed, it is separately added into methanol
40ml, ultrasonic (35KHz, 280W) extracts 60min, filters, and filtrate volatilizes, and residue adds methanol and dissolves, be settled to 10ml and use
0.22 μm filtering with microporous membrane, takes subsequent filtrate, to obtain final product.
(2) preparation of reference substance solution: take Calycosin-7-O-BETA-D-glucoside reference substance, ononin reference substance, 9,10-dimethoxy
Lignum pterocarpi indici alkane-3-O-β-D glucopyranoside reference substance, calycosin reference substance, astragaloside reference substance, Radix Astragali saponin III are right
According to product, formononetin reference substance, astragaloside I reference substance in right amount, accurately weighed, add methanol and make every 1ml respectively containing Mao Ruiyi Huang
Ketoside 0.2mg, ononin 0.05mg, 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-β-D glucopyranoside 0.08mg, Mao Ruiyi
Flavone 0.2mg, astragaloside 1.4mg, Radix Astragali saponin III 1.2mg, formononetin 0.05mg, astragaloside I 1.2mg mixing molten
Liquid, to obtain final product.
The most the places of production and the analysis of batch Radix Astragali finger printing
(1) selection of reference colored spectral peak: by the HPLC chromatogram collection of illustrative plates of 13 different batches Radixs Astragali is carried out system thinking,
At discovery 49.3min, the separating degree of chromatographic peak (peak 16) is good, content is high and stablizes, and theoretical cam curve is no less than 70000, therefore selects
Take peak 16 as reference colored spectral peak.
(2) determination of total chromatographic peak: by calculating, comparing each chromatograph in 13 different batches Radix Astragali sample finger printing
The relative retention time at the peak (ratio of each chromatographic peak retention time and the retention time of wherein reference colored spectral peak in the most same spectrogram
Value), wherein 26 chromatographic peaks are confirmed as total chromatographic peak.
(3) confirm the composition of the total chromatographic peak of part: by the comparison with reference substance retention time, determine that peak 10 is hair stamen
Isoflavone glucoside；Peak 12 is ononin；Peak 13 is 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-β-D glucopyranoside；
Peak 16 is calycosin；Peak 20 is astragaloside；Peak 21 is Radix Astragali saponin III；Peak 22 is formononetin；Peak 26 is Radix Astragali soap
Glycosides I.
As shown in figure 12, batch finger printing is as shown in figure 13 prolifically for gained reference substance finger printing, standard fingerprint figure
Spectrum is as shown in figure 14.
Embodiment 3
Radix Astragali HPLC fingerprint spectrum method is investigated
1. precision test
Take the same place of production batch Radix Astragali (place of production is Gansu, and lot number is 130313) need testing solution, continuous sample introduction 6 times, examine
The relative retention time of observation of complexion spectral peak, the concordance of relative peak area ratio, thus investigate the precision of instrument, the results are shown in Table 5,
Shown in table 6.Result shows, the relative retention time of each total chromatographic peak and the RSD value of relative peak area ratio all 3% with
In, show that the precision of instrument is good.
Table 5 Radix Astragali essenc density test relative retention time
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | RSD (%) |
| 1 | 0.104 | 0.104 | 0.104 | 0.104 | 0.104 | 0.104 | 0.132 |
| 2 | 0.142 | 0.142 | 0.142 | 0.142 | 0.141 | 0.141 | 0.104 |
| 3 | 0.159 | 0.159 | 0.159 | 0.159 | 0.159 | 0.159 | 0.077 |
| 4 | 0.202 | 0.202 | 0.202 | 0.202 | 0.201 | 0.201 | 0.096 |
| 5 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.082 |
| 6 | 0.345 | 0.345 | 0.344 | 0.344 | 0.344 | 0.344 | 0.173 |
| 7 | 0.441 | 0.440 | 0.440 | 0.440 | 0.439 | 0.439 | 0.134 |
| 8 | 0.457 | 0.457 | 0.456 | 0.456 | 0.456 | 0.456 | 0.106 |
| 9 | 0.487 | 0.487 | 0.487 | 0.487 | 0.487 | 0.487 | 0.011 |
| 10 | 0.551 | 0.550 | 0.550 | 0.550 | 0.550 | 0.550 | 0.019 |
| 11 | 0.643 | 0.643 | 0.643 | 0.643 | 0.643 | 0.643 | 0.023 |
| 12 | 0.812 | 0.813 | 0.812 | 0.812 | 0.812 | 0.812 | 0.025 |
| 13 | 0.895 | 0.895 | 0.895 | 0.895 | 0.895 | 0.895 | 0.023 |
| 14 | 0.920 | 0.920 | 0.920 | 0.920 | 0.920 | 0.920 | 0.018 |
| 15 | 0.941 | 0.941 | 0.940 | 0.940 | 0.940 | 0.940 | 0.017 |
| 16 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.000 |
| 17 | 1.146 | 1.145 | 1.145 | 1.145 | 1.145 | 1.145 | 0.020 |
| 18 | 1.212 | 1.211 | 1.211 | 1.211 | 1.212 | 1.213 | 0.063 |
| 19 | 1.293 | 1.293 | 1.293 | 1.293 | 1.293 | 1.293 | 0.022 |
| 20 | 1.323 | 1.322 | 1.323 | 1.322 | 1.322 | 1.322 | 0.022 |
| 21 | 1.346 | 1.345 | 1.346 | 1.345 | 1.345 | 1.345 | 0.025 |
| 22 | 1.418 | 1.417 | 1.417 | 1.417 | 1.417 | 1.417 | 0.022 |
| 23 | 1.450 | 1.449 | 1.449 | 1.449 | 1.449 | 1.449 | 0.031 |
| 24 | 1.475 | 1.474 | 1.474 | 1.474 | 1.473 | 1.474 | 0.035 |
| 25 | 1.520 | 1.519 | 1.520 | 1.519 | 1.519 | 1.519 | 0.036 |
| 26 | 1.666 | 1.665 | 1.666 | 1.665 | 1.665 | 1.665 | 0.027 |
Table 6 Radix Astragali essenc density test relative peak area
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | RSD (%) |
| 1 | 0.188 | 0.189 | 0.189 | 0.187 | 0.186 | 0.188 | 0.697 |
| 2 | 0.621 | 0.627 | 0.619 | 0.631 | 0.614 | 0.618 | 0.997 |
| 3 | 0.138 | 0.136 | 0.136 | 0.138 | 0.135 | 0.135 | 0.939 |
| 4 | 0.078 | 0.078 | 0.078 | 0.078 | 0.078 | 0.078 | 0.287 |
| 5 | 0.072 | 0.071 | 0.072 | 0.072 | 0.071 | 0.072 | 0.802 |
| 6 | 0.353 | 0.345 | 0.349 | 0.346 | 0.344 | 0.345 | 1.062 |
| 7 | 0.151 | 0.148 | 0.151 | 0.149 | 0.148 | 0.150 | 0.934 |
| 8 | 0.132 | 0.134 | 0.134 | 0.132 | 0.132 | 0.135 | 0.988 |
| 9 | 0.005 | 0.004 | 0.005 | 0.004 | 0.004 | 0.004 | 2.033 |
| 10 | 0.322 | 0.319 | 0.317 | 0.320 | 0.317 | 0.323 | 0.787 |
| 11 | 0.080 | 0.079 | 0.080 | 0.079 | 0.078 | 0.079 | 0.655 |
| 12 | 0.061 | 0.060 | 0.060 | 0.060 | 0.060 | 0.060 | 0.600 |
| 13 | 0.211 | 0.208 | 0.212 | 0.209 | 0.210 | 0.208 | 0.751 |
| 14 | 0.063 | 0.063 | 0.062 | 0.062 | 0.065 | 0.063 | 1.805 |
| 15 | 0.665 | 0.658 | 0.665 | 0.655 | 0.659 | 0.664 | 0.617 |
| 16 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.000 |
| 17 | 0.122 | 0.121 | 0.124 | 0.122 | 0.122 | 0.122 | 0.936 |
| 18 | 0.032 | 0.033 | 0.032 | 0.031 | 0.033 | 0.032 | 1.974 |
| 19 | 0.112 | 0.116 | 0.116 | 0.117 | 0.116 | 0.116 | 1.525 |
| 20 | 0.136 | 0.138 | 0.140 | 0.138 | 0.140 | 0.141 | 1.313 |
| 21 | 0.054 | 0.054 | 0.054 | 0.054 | 0.053 | 0.054 | 0.546 |
| 22 | 0.362 | 0.364 | 0.363 | 0.367 | 0.372 | 0.367 | 0.944 |
| 23 | 0.203 | 0.200 | 0.202 | 0.201 | 0.199 | 0.200 | 0.720 |
| 24 | 1.173 | 1.177 | 1.174 | 1.183 | 1.178 | 1.181 | 0.317 |
| 25 | 2.880 | 2.873 | 2.881 | 2.873 | 2.861 | 2.881 | 0.274 |
| 26 | 0.027 | 0.027 | 0.027 | 0.026 | 0.027 | 0.027 | 1.308 |
2. stability test
Take the need testing solution of the same place of production batch Radix Astragali (place of production is Gansu, and lot number is 130313), respectively 0h, 2h,
4h, 8h, 12h, 24h detect, and to investigate the stability of sample, the results are shown in Table 7, shown in table 8.Result shows, each total color
The relative retention time of spectral peak and the RSD value of relative peak area ratio are respectively less than 3%, illustrate that sample solution is the most steady in 24h
Fixed.
Table 7 Radix Astragali stability test relative retention time
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | RSD (%) |
| 1 | 0.104 | 0.104 | 0.104 | 0.104 | 0.104 | 0.104 | 0.050 |
| 2 | 0.142 | 0.142 | 0.142 | 0.142 | 0.142 | 0.142 | 0.058 |
| 3 | 0.159 | 0.159 | 0.159 | 0.159 | 0.159 | 0.159 | 0.034 |
| 4 | 0.202 | 0.202 | 0.202 | 0.202 | 0.202 | 0.202 | 0.060 |
| 5 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.037 |
| 6 | 0.344 | 0.344 | 0.344 | 0.344 | 0.344 | 0.344 | 0.040 |
| 7 | 0.433 | 0.433 | 0.433 | 0.433 | 0.433 | 0.433 | 0.037 |
| 8 | 0.454 | 0.454 | 0.453 | 0.454 | 0.454 | 0.454 | 0.047 |
| 9 | 0.487 | 0.487 | 0.487 | 0.487 | 0.487 | 0.487 | 0.034 |
| 10 | 0.552 | 0.552 | 0.552 | 0.553 | 0.552 | 0.552 | 0.020 |
| 11 | 0.646 | 0.646 | 0.646 | 0.646 | 0.645 | 0.645 | 0.037 |
| 12 | 0.816 | 0.816 | 0.816 | 0.816 | 0.816 | 0.816 | 0.037 |
| 13 | 0.898 | 0.898 | 0.898 | 0.899 | 0.898 | 0.898 | 0.013 |
| 14 | 0.924 | 0.924 | 0.924 | 0.924 | 0.924 | 0.924 | 0.014 |
| 15 | 0.944 | 0.944 | 0.944 | 0.944 | 0.944 | 0.944 | 0.009 |
| 16 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.000 |
| 17 | 1.149 | 1.149 | 1.149 | 1.149 | 1.149 | 1.149 | 0.010 |
| 18 | 1.218 | 1.218 | 1.217 | 1.217 | 1.217 | 1.217 | 0.022 |
| 19 | 1.290 | 1.290 | 1.289 | 1.289 | 1.290 | 1.290 | 0.010 |
| 20 | 1.327 | 1.327 | 1.327 | 1.327 | 1.327 | 1.327 | 0.007 |
| 21 | 1.350 | 1.350 | 1.350 | 1.350 | 1.350 | 1.350 | 0.007 |
| 22 | 1.419 | 1.418 | 1.418 | 1.418 | 1.418 | 1.418 | 0.014 |
| 23 | 1.455 | 1.455 | 1.455 | 1.454 | 1.455 | 1.455 | 0.012 |
| 24 | 1.479 | 1.479 | 1.479 | 1.479 | 1.479 | 1.479 | 0.010 |
| 25 | 1.525 | 1.525 | 1.525 | 1.524 | 1.525 | 1.525 | 0.007 |
| 26 | 1.671 | 1.671 | 1.671 | 1.671 | 1.671 | 1.671 | 0.005 |
Table 8 Radix Astragali stability test relative peak area
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | RSD (%) |
| 1 | 0.215 | 0.212 | 0.211 | 0.212 | 0.213 | 0.212 | 0.585 |
| 2 | 0.753 | 0.739 | 0.736 | 0.736 | 0.738 | 0.737 | 0.880 |
| 3 | 0.137 | 0.139 | 0.139 | 0.138 | 0.137 | 0.135 | 0.977 |
| 4 | 0.034 | 0.034 | 0.034 | 0.034 | 0.034 | 0.034 | 0.545 |
| 5 | 0.129 | 0.129 | 0.127 | 0.131 | 0.128 | 0.127 | 1.231 |
| 6 | 0.372 | 0.368 | 0.367 | 0.371 | 0.372 | 0.370 | 0.595 |
| 7 | 0.140 | 0.141 | 0.141 | 0.142 | 0.142 | 0.142 | 0.538 |
| 8 | 0.187 | 0.186 | 0.185 | 0.186 | 0.189 | 0.184 | 1.000 |
| 9 | 0.430 | 0.423 | 0.421 | 0.421 | 0.423 | 0.420 | 0.896 |
| 10 | 0.281 | 0.279 | 0.277 | 0.281 | 0.280 | 0.278 | 0.592 |
| 11 | 0.067 | 0.066 | 0.065 | 0.067 | 0.065 | 0.066 | 1.724 |
| 12 | 0.081 | 0.081 | 0.080 | 0.081 | 0.081 | 0.081 | 0.495 |
| 13 | 0.178 | 0.176 | 0.174 | 0.176 | 0.175 | 0.177 | 0.695 |
| 14 | 0.059 | 0.059 | 0.059 | 0.059 | 0.060 | 0.060 | 1.289 |
| 15 | 0.528 | 0.531 | 0.539 | 0.538 | 0.546 | 0.542 | 1.273 |
| 16 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.000 |
| 17 | 0.121 | 0.122 | 0.120 | 0.123 | 0.124 | 0.122 | 1.155 |
| 18 | 0.021 | 0.021 | 0.021 | 0.021 | 0.021 | 0.021 | 0.401 |
| 19 | 0.076 | 0.077 | 0.075 | 0.076 | 0.075 | 0.077 | 0.986 |
| 20 | 0.150 | 0.149 | 0.148 | 0.149 | 0.150 | 0.149 | 0.583 |
| 21 | 0.055 | 0.055 | 0.054 | 0.054 | 0.054 | 0.054 | 0.625 |
| 22 | 0.363 | 0.366 | 0.367 | 0.366 | 0.370 | 0.366 | 0.639 |
| 23 | 0.162 | 0.163 | 0.160 | 0.161 | 0.163 | 0.162 | 0.624 |
| 24 | 1.267 | 1.266 | 1.251 | 1.258 | 1.257 | 1.257 | 0.464 |
| 25 | 2.960 | 2.938 | 2.923 | 2.933 | 2.945 | 2.932 | 0.435 |
| 26 | 0.022 | 0.021 | 0.021 | 0.021 | 0.021 | 0.021 | 1.623 |
3. replica test
Take the same place of production batch Radix Astragali (place of production is Gansu, and lot number is 130313) 6 parts, prepare respectively need testing solution and by
Detect according to selected chromatographic condition, the results are shown in Table 9, shown in table 10.Visible, the relative retention time of each total chromatographic peak and
Relative peak area RSD value is respectively less than 3%, shows that repeatability is good.
Table 9 Radix Astragali replica test relative retention time
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | RSD (%) |
| 1 | 0.104 | 0.104 | 0.104 | 0.104 | 0.104 | 0.104 | 0.073 |
| 2 | 0.142 | 0.142 | 0.142 | 0.142 | 0.142 | 0.142 | 0.053 |
| 3 | 0.159 | 0.159 | 0.159 | 0.159 | 0.159 | 0.159 | 0.032 |
| 4 | 0.202 | 0.202 | 0.202 | 0.202 | 0.202 | 0.202 | 0.073 |
| 5 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.241 | 0.051 |
| 6 | 0.342 | 0.342 | 0.342 | 0.342 | 0.342 | 0.342 | 0.048 |
| 7 | 0.432 | 0.432 | 0.432 | 0.432 | 0.432 | 0.432 | 0.052 |
| 8 | 0.452 | 0.452 | 0.452 | 0.452 | 0.452 | 0.452 | 0.046 |
| 9 | 0.484 | 0.484 | 0.484 | 0.484 | 0.484 | 0.484 | 0.031 |
| 10 | 0.552 | 0.552 | 0.552 | 0.552 | 0.552 | 0.552 | 0.024 |
| 11 | 0.646 | 0.646 | 0.646 | 0.645 | 0.645 | 0.646 | 0.029 |
| 12 | 0.816 | 0.816 | 0.816 | 0.816 | 0.816 | 0.816 | 0.030 |
| 13 | 0.898 | 0.898 | 0.898 | 0.898 | 0.897 | 0.898 | 0.024 |
| 14 | 0.924 | 0.924 | 0.924 | 0.923 | 0.923 | 0.923 | 0.027 |
| 15 | 0.944 | 0.944 | 0.944 | 0.943 | 0.943 | 0.943 | 0.021 |
| 16 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.000 |
| 17 | 1.147 | 1.147 | 1.147 | 1.147 | 1.147 | 1.147 | 0.007 |
| 18 | 1.218 | 1.218 | 1.218 | 1.218 | 1.218 | 1.218 | 0.016 |
| 19 | 1.290 | 1.290 | 1.290 | 1.290 | 1.290 | 1.290 | 0.017 |
| 20 | 1.325 | 1.325 | 1.325 | 1.325 | 1.325 | 1.325 | 0.007 |
| 21 | 1.349 | 1.348 | 1.348 | 1.348 | 1.348 | 1.348 | 0.020 |
| 22 | 1.418 | 1.418 | 1.418 | 1.418 | 1.419 | 1.419 | 0.015 |
| 23 | 1.453 | 1.453 | 1.453 | 1.453 | 1.453 | 1.453 | 0.009 |
| 24 | 1.477 | 1.477 | 1.477 | 1.477 | 1.477 | 1.477 | 0.007 |
| 25 | 1.523 | 1.523 | 1.523 | 1.523 | 1.523 | 1.523 | 0.012 |
| 26 | 1.671 | 1.670 | 1.670 | 1.670 | 1.671 | 1.671 | 0.027 |
Table 10 Radix Astragali replica test relative peak area
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | RSD (%) |
| 1 | 0.200 | 0.199 | 0.200 | 0.203 | 0.194 | 0.193 | 1.980 |
| 2 | 0.704 | 0.701 | 0.707 | 0.717 | 0.685 | 0.684 | 1.891 |
| 3 | 0.125 | 0.125 | 0.125 | 0.127 | 0.122 | 0.121 | 1.810 |
| 4 | 0.062 | 0.062 | 0.062 | 0.063 | 0.060 | 0.061 | 1.774 |
| 5 | 0.087 | 0.087 | 0.088 | 0.088 | 0.085 | 0.084 | 1.936 |
| 6 | 0.304 | 0.303 | 0.305 | 0.310 | 0.295 | 0.295 | 1.914 |
| 7 | 0.055 | 0.054 | 0.055 | 0.055 | 0.053 | 0.053 | 1.716 |
| 8 | 0.174 | 0.173 | 0.175 | 0.177 | 0.169 | 0.169 | 1.931 |
| 9 | 0.456 | 0.453 | 0.454 | 0.448 | 0.460 | 0.441 | 1.441 |
| 10 | 0.265 | 0.264 | 0.266 | 0.270 | 0.257 | 0.257 | 1.928 |
| 11 | 0.066 | 0.066 | 0.066 | 0.067 | 0.065 | 0.064 | 1.843 |
| 12 | 0.074 | 0.074 | 0.074 | 0.075 | 0.072 | 0.072 | 1.877 |
| 13 | 0.165 | 0.164 | 0.165 | 0.168 | 0.160 | 0.160 | 1.883 |
| 14 | 0.062 | 0.062 | 0.062 | 0.062 | 0.060 | 0.060 | 1.593 |
| 15 | 0.547 | 0.545 | 0.552 | 0.556 | 0.565 | 0.565 | 1.593 |
| 16 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 1.000 | 0.000 |
| 17 | 0.122 | 0.120 | 0.124 | 0.119 | 0.121 | 0.121 | 1.392 |
| 18 | 0.027 | 0.027 | 0.027 | 0.027 | 0.026 | 0.026 | 1.946 |
| 19 | 0.076 | 0.076 | 0.076 | 0.077 | 0.074 | 0.074 | 1.948 |
| 20 | 0.136 | 0.136 | 0.137 | 0.139 | 0.133 | 0.133 | 1.883 |
| 21 | 0.047 | 0.046 | 0.047 | 0.047 | 0.045 | 0.045 | 1.878 |
| 22 | 0.368 | 0.363 | 0.363 | 0.368 | 0.366 | 0.366 | 0.635 |
| 23 | 0.160 | 0.160 | 0.161 | 0.163 | 0.156 | 0.156 | 1.966 |
| 24 | 1.281 | 1.296 | 1.276 | 1.308 | 1.249 | 1.244 | 1.970 |
| 25 | 3.002 | 3.013 | 3.010 | 3.095 | 2.961 | 2.957 | 1.656 |
| 26 | 0.023 | 0.023 | 0.023 | 0.023 | 0.022 | 0.022 | 1.863 |
The most the places of production, the similarity evaluations of multiple batch Radix Astragali finger printing
Use similarity evaluation (2012.130723 version) to different batches Radix Astragali HPLC
Spectrogram carries out fingerprint similarity calculating, the results are shown in Table 11.
Table 11 different sources batch Radix Astragali fingerprint similarity result of calculation
| S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | S9 | S10 | S11 | S12 | S13 | R* | |
| S1 | 1.000 | |||||||||||||
| S2 | 0.930 | 1.000 |
| S3 | 0.958 | 0.963 | 1.000 | |||||||||||
| S4 | 0.860 | 0.895 | 0.922 | 1.000 | ||||||||||
| S5 | 0.869 | 0.851 | 0.920 | 0.961 | 1.000 | |||||||||
| S6 | 0.800 | 0.895 | 0.850 | 0.903 | 0.817 | 1.000 | ||||||||
| S7 | 0.756 | 0.855 | 0.812 | 0.905 | 0.802 | 0.933 | 1.000 | |||||||
| S8 | 0.887 | 0.869 | 0.878 | 0.882 | 0.864 | 0.847 | 0.830 | 1.000 | ||||||
| S9 | 0.815 | 0.785 | 0.894 | 0.852 | 0.866 | 0.705 | 0.702 | 0.697 | 1.000 | |||||
| S10 | 0.806 | 0.895 | 0.850 | 0.868 | 0.813 | 0.884 | 0.885 | 0.874 | 0.675 | 1.000 | ||||
| S11 | 0.777 | 0.889 | 0.859 | 0.908 | 0.831 | 0.948 | 0.933 | 0.856 | 0.741 | 0.913 | 1.000 | |||
| S12 | 0.909 | 0.914 | 0.948 | 0.955 | 0.953 | 0.874 | 0.876 | 0.918 | 0.850 | 0.884 | 0.892 | 1.000 | ||
| S13 | 0.847 | 0.836 | 0.910 | 0.948 | 0.959 | 0.807 | 0.828 | 0.869 | 0.885 | 0.816 | 0.847 | 0.963 | 1.000 | |
| R* | 0.909 | 0.942 | 0.962 | 0.977 | 0.948 | 0.922 | 0.915 | 0.919 | 0.871 | 0.918 | 0.940 | 0.980 | 0.953 | 1.0 00 |
Table note: * R is standard finger-print；Time window width 0.1, intercepts 5--85 minute spectrogram, full peak match
The most numbered Huang of 9,10 is calculated by similarity evaluation 2012.130723 version
Stilbene differs relatively big with the Radix Astragali similarity of other batches, and remaining 11 batch Radix Astragali is equal with the similarity result of standard finger-print
More than 0.9.
It addition, take the Radix Astragali, the Radix Astragali of numbered 12, Radix Astagali control medicinal material and the Radix Astragali of numbered 6 respectively
Control medicinal material carries out fingerprint map analyzing, and finger printing comparison diagram is shown in Figure 15 (S1: Radix Astragali of numbered 12；S2: numbered 6
The Radix Astragali；S3: Radix Astagali control medicinal material；S4: Radix Astragali control medicinal material), Similarity Measure the results are shown in Table 12.
12 two batch Radixs Astragali of table and Radix Astagali and Radix Astragali fingerprint similarity result of calculation table
| The Radix Astragali of numbered 12 | The Radix Astragali of numbered 6 | Radix Astagali | Radix Astragali | |
| The Radix Astragali of numbering 12 | 1.000 | |||
| The Radix Astragali of numbering 6 | 0.894 | 1.000 | ||
| Radix Astagali | 0.882 | 0.727 | 1.000 | |
| Radix Astragali | 0.710 | 0.747 | 0.699 | 1.000 |
Compared by finger printing and can be seen that, the Radix Astragali of two batches and Radix Astagali and the total chromatographic peak one of Radix Astragali
Cause, but each peak-to-peak area ratio has different, characteristic peak can determine whether that they are certified products medical material.By similarity result the most not
The source that can judge two batch Radixs Astragali is Radix Astagali or Radix Astragali.
Claims (6)
1. an assay method for Radix Astragali finger printing, including:
(1) powder in the Radix Astragali is taken, accurately weighed, add methanol, 35KHz supersound extraction 60min, filter, filtrate volatilizes, and residue adds first
Alcohol dissolves, constant volume with filtering with microporous membrane, takes subsequent filtrate, obtains need testing solution；
(2) calycosin glucoside reference substance, ononin reference substance, 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-β-D are taken
Glucopyranoside reference substance, calycosin reference substance, astragaloside reference substance, Radix Astragali saponin III reference substance, formononetin
Reference substance, astragaloside I reference substance, accurately weighed, it is separately added into methanol and makes reference substance solution；
(3) precision draws need testing solution and reference substance solution respectively, injects high performance liquid chromatograph and measures, obtains Radix Astragali fingerprint
Collection of illustrative plates；Wherein, chromatographic condition is: chromatographic column: Athena C18-WP chromatographic column；Column temperature: 30 DEG C；Detection wavelength: 208nm；Sample introduction
Volume: 10 μ l；Flow velocity: 1ml min-1；Flowing phase: mobile phase A is water, and Mobile phase B is acetonitrile；Gradient elution program: 0～
20min:5%B → 20%B, 20～70min:20%B → 40%B, 70～85min:40%B → 60%B.
The assay method of a kind of Radix Astragali finger printing the most according to claim 1, it is characterised in that: in described step (1)
The Radix Astragali in the ratio of powder and methanol be 1g:20ml.
The assay method of a kind of Radix Astragali finger printing the most according to claim 1, it is characterised in that: in described step (1)
The aperture of microporous filter membrane be 0.22 μm.
The assay method of a kind of Radix Astragali finger printing the most according to claim 1, it is characterised in that: in described step (2)
The ratio of reference substance and methanol be 0.05～1.5mg:1ml.
The assay method of a kind of Radix Astragali finger printing the most according to claim 1, it is characterised in that: in described step (2)
Reference substance solution particularly as follows: every 1ml solution containing Calycosin-7-O-BETA-D-glucoside 0.2mg；Every 1ml contains the molten of ononin 0.05mg
Liquid；Every 1ml solution containing 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-β-D glucopyranoside 0.08mg；Every 1ml is yellow containing Mao Ruiyi
The solution of ketone 0.2mg；Every 1ml solution containing astragaloside 1.4mg；Every 1ml solution containing Radix Astragali saponin III 1.2mg；Every 1ml contains
The solution of formononetin 0.05mg；Every 1ml solution containing astragaloside I 1.2mg.
The assay method of a kind of Radix Astragali finger printing the most according to claim 1, it is characterised in that: in described step (3)
Radix Astragali finger printing in 26 chromatographic peaks be confirmed as total chromatographic peak, by the comparison with reference substance retention time, determine
Peak 10 is calycosin glucoside；Peak 12 is ononin；Peak 13 is 9,10-dimethoxy Lignum pterocarpi indici alkane-3-O-β-D Fructus Vitis viniferae
Pyranoside；Peak 16 is calycosin；Peak 20 is astragaloside；Peak 21 is Radix Astragali saponin III；Peak 22 is formononetin；Peak 26
For astragaloside I.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410857192.8A CN104569199B (en) | 2014-12-30 | 2014-12-30 | A kind of assay method of Radix Astragali finger printing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410857192.8A CN104569199B (en) | 2014-12-30 | 2014-12-30 | A kind of assay method of Radix Astragali finger printing |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104569199A CN104569199A (en) | 2015-04-29 |
| CN104569199B true CN104569199B (en) | 2016-08-24 |
Family
ID=53085760
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410857192.8A Active CN104569199B (en) | 2014-12-30 | 2014-12-30 | A kind of assay method of Radix Astragali finger printing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104569199B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792652B (en) * | 2015-05-02 | 2017-07-25 | 浙江大学 | A kind of Milkvetch Root multiple index quick detecting method |
| CN105021751B (en) * | 2015-06-30 | 2016-09-07 | 广州市香雪制药股份有限公司 | The content assaying method of Multiple components in Yupingfeng preparation |
| CN106324167B (en) * | 2015-07-06 | 2020-04-03 | 天津天士力现代中药资源有限公司 | Method for determining flavonoid components in astragalus extract by UPLC |
| CN107045030A (en) * | 2017-04-17 | 2017-08-15 | 广西壮族自治区梧州食品药品检验所 | A kind of method of calycosin glucoside in ASE methods extraction Radix Astragali |
| CN106950324B (en) * | 2017-05-18 | 2019-06-28 | 山西大学 | A kind of Radix Astragali builds the construction method of middle ball finger-print |
| CN107907619A (en) * | 2017-10-25 | 2018-04-13 | 中山市中智药业集团有限公司 | A kind of blood concentration quantitative analysis method of radix astragali broken wall medicine materical crude slice active ingredient |
| CN108042603A (en) * | 2018-01-16 | 2018-05-18 | 山西大学 | Radix Astragali flavone extract is preparing the application in treating nephrotic syndrome drug |
| CN109828062A (en) * | 2019-04-10 | 2019-05-31 | 山西大学 | A kind of Radix Astragali builds the measuring method of 18 kinds of component contents in middle ball |
| CN111024871A (en) * | 2019-12-06 | 2020-04-17 | 中国药科大学 | Radix astragali general cargo and goods selection grading marker, detection kit and application thereof |
| CN112834650B (en) * | 2021-01-12 | 2022-10-21 | 湖南天劲制药有限责任公司 | Quality detection method of bone-strengthening and blood-generating oral liquid |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857380A (en) * | 2006-03-22 | 2006-11-08 | 四川科伦药业股份有限公司 | Method for detecting medicine material astragalus quality |
| CN101327246A (en) * | 2008-07-29 | 2008-12-24 | 四川百利药业有限责任公司 | Radix astragali medicinal materials, intermediate body and method for testing fingerprint of formulation thereof as well as standard fingerprint |
| CN102353729A (en) * | 2006-03-22 | 2012-02-15 | 四川科伦药业股份有限公司 | Quality detection method of radix astragali medicinal material |
| CN102879486A (en) * | 2012-09-05 | 2013-01-16 | 山东大学 | Method for screening traditional Chinese medicine effect related ingredients and model building method |
| CN103149300A (en) * | 2013-03-05 | 2013-06-12 | 贵州师范大学 | Measurement method of radix astragali granule fingerprint and characteristic fingerprint thereof |
| CN103969355A (en) * | 2013-02-02 | 2014-08-06 | 西安世纪盛康药业有限公司 | Identification method for fingerprint spectrum of astragalus medicinal material |
| CN104181186A (en) * | 2014-09-03 | 2014-12-03 | 山西大学 | Construction method of <1>H-NMR fingerprint of radix astragali injection |
| CN104198600A (en) * | 2014-07-31 | 2014-12-10 | 甘肃中天药业有限责任公司 | Method for detecting radix astragali |
-
2014
- 2014-12-30 CN CN201410857192.8A patent/CN104569199B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1857380A (en) * | 2006-03-22 | 2006-11-08 | 四川科伦药业股份有限公司 | Method for detecting medicine material astragalus quality |
| CN102353729A (en) * | 2006-03-22 | 2012-02-15 | 四川科伦药业股份有限公司 | Quality detection method of radix astragali medicinal material |
| CN101327246A (en) * | 2008-07-29 | 2008-12-24 | 四川百利药业有限责任公司 | Radix astragali medicinal materials, intermediate body and method for testing fingerprint of formulation thereof as well as standard fingerprint |
| CN102879486A (en) * | 2012-09-05 | 2013-01-16 | 山东大学 | Method for screening traditional Chinese medicine effect related ingredients and model building method |
| CN103969355A (en) * | 2013-02-02 | 2014-08-06 | 西安世纪盛康药业有限公司 | Identification method for fingerprint spectrum of astragalus medicinal material |
| CN103149300A (en) * | 2013-03-05 | 2013-06-12 | 贵州师范大学 | Measurement method of radix astragali granule fingerprint and characteristic fingerprint thereof |
| CN104198600A (en) * | 2014-07-31 | 2014-12-10 | 甘肃中天药业有限责任公司 | Method for detecting radix astragali |
| CN104181186A (en) * | 2014-09-03 | 2014-12-03 | 山西大学 | Construction method of <1>H-NMR fingerprint of radix astragali injection |
Non-Patent Citations (3)
| Title |
|---|
| Liu, Xiao-hua;Zhao, Liang-gong;Liang, Jing.Component analysis and structure identification of active substances for anti-gastric ulcer effects in Radix Astragali by liquid chromatography and tandem mass spectrometry.《JOURNAL OF CHROMATOGRAPHY》.2014,第960卷 * |
| 膜荚黄芪与蒙古黄芪化学成分的高效液相色谱-质谱研究;李锐 等;《分析化学》;20051231;第33卷(第12期);1676-1680 * |
| 蒙古黄芪主产区2 种不同种植模式黄芪药材的质量比较;杜国军 等;《中草药》;20131215;第44卷(第23期);3386-3393 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104569199A (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104569199B (en) | A kind of assay method of Radix Astragali finger printing | |
| CN101850070B (en) | Detection method for Chinese medicament Tangcao tablets | |
| CN104713956B (en) | Method for determining fingerprint chromatography of radix astragali and ligusticum wallichii extract products | |
| CN102749348B (en) | Method for identifying active components in medicinal plant | |
| CN101732607B (en) | Method for detecting quality of huaqi Chinese medicinal preparation | |
| CN108226313A (en) | In glutinous rehmannia while methods of glycosides measure and fingerprint map construction method | |
| CN103800523A (en) | Method for preparing anti-virus traditional Chinese medicinal composition and finger-print detection method | |
| CN106442789A (en) | Establishment and active component quantitative analysis methods of compound Xuezhining extract fingerprint map | |
| CN106370763B (en) | UPLC method for detecting kudzu root, kudzu root extract and kudzu root preparation component | |
| Yin et al. | Quality analysis of different specification grades of Astragalus membranaceus var. mongholicus (Huangqi) from Hunyuan, Shanxi | |
| CN103592391B (en) | The content assaying method of Specnuezhenide in Zhenqi Fuzheng prepn | |
| Qi et al. | HPLC fingerprint and LC-TOF-MS analysis on extract from roots of Gentiana macrophylla | |
| CN102488819A (en) | Preparing method for daylily flower extract | |
| CN106226435A (en) | A kind of fingerprint atlas detection method of Yixinshu tablet saponin component | |
| CN106596426A (en) | Detection method applied in gynostemma total saponin extraction and separation process | |
| CN107643343B (en) | HPLC fingerprint spectrum determination method of Yunv Jian standard soup | |
| CN108872417A (en) | A kind of construction method and HPLC finger-print of finger-print | |
| CN103969355B (en) | A kind of discrimination method of the finger printing of Milkvetch Root | |
| CN116124937A (en) | Construction method and application of finger print of Zhuanyanming tablet | |
| CN102628838B (en) | Fujian herb cusia HPLC fingerprint construction method | |
| CN102008541B (en) | Method for simultaneously detecting three main active ingredients in sugar-free type compound wintercreeper preparation | |
| Liu et al. | Simultaneous determination of ginsenosides and lignans in Sheng-mai injection by ultra-performance liquid chromatography with diode array detection | |
| CN103301291A (en) | Qi-replenishing and blood-enriching longan soup as well as preparation method and quality detection method thereof | |
| CN106442801A (en) | Method of simultaneously measuring content of 11 active ingredients in folium isatidis | |
| CN103115976A (en) | Method for measuring content of in-blood chemical component-allantoin in medicinal material-psammosilene tunicoides |
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 | ||
| CP01 | Change in the name or title of a patent holder |
Address after: Room 3301-3302, No. 1027, Changning Road, Changning District, Shanghai 200050 Patentee after: Shanghai Baiyang Pharmaceutical Co.,Ltd. Address before: Room 3301-3302, No. 1027, Changning Road, Changning District, Shanghai 200050 Patentee before: Shanghai Sundise Chinese Medicine Technology Development Co.,Ltd. |
|
| CP01 | Change in the name or title of a patent holder |