CN107793465A - A kind of extracting method and the application of ginsenoside Rg 5 and ginsenoside Rk1 - Google Patents
A kind of extracting method and the application of ginsenoside Rg 5 and ginsenoside Rk1 Download PDFInfo
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- CN107793465A CN107793465A CN201710865039.3A CN201710865039A CN107793465A CN 107793465 A CN107793465 A CN 107793465A CN 201710865039 A CN201710865039 A CN 201710865039A CN 107793465 A CN107793465 A CN 107793465A
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- ginsenoside
- organic solvent
- containing water
- water elution
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- KWDWBAISZWOAHD-MHOSXIPRSA-N (2s,3r,4s,5s,6r)-2-[(2r,3r,4s,5s,6r)-4,5-dihydroxy-6-(hydroxymethyl)-2-[[(3s,5r,8r,9r,10r,12r,13r,14r,17s)-12-hydroxy-4,4,8,10,14-pentamethyl-17-(6-methylhepta-1,5-dien-2-yl)-2,3,5,6,7,9,11,12,13,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-3-yl]oxy]o Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1CC[C@]2(C)[C@H]3C[C@@H](O)[C@H]4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@@H]4C(=C)CCC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O KWDWBAISZWOAHD-MHOSXIPRSA-N 0.000 title claims abstract description 118
- KWDWBAISZWOAHD-UHFFFAOYSA-N Ginsenoside Rk1 Natural products CC(C)=CCCC(=C)C1CCC(C2(CCC3C4(C)C)C)(C)C1C(O)CC2C3(C)CCC4OC1OC(CO)C(O)C(O)C1OC1OC(CO)C(O)C(O)C1O KWDWBAISZWOAHD-UHFFFAOYSA-N 0.000 title claims abstract description 118
- NJUXRKMKOFXMRX-RNCAKNGISA-N Ginsenoside Rg5 Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1CC[C@]2(C)[C@H]3C[C@@H](O)[C@H]4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@@H]4C(/C)=C/CC=C(C)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O NJUXRKMKOFXMRX-RNCAKNGISA-N 0.000 title claims abstract description 117
- NJUXRKMKOFXMRX-AXUZFSSLSA-N ginsenoside Rg5 Natural products CC(=CCC=C(C)[C@H]1CC[C@]2(C)[C@@H]1[C@H](O)C[C@@H]3[C@@]4(C)CC[C@@H](O[C@H]5O[C@H](CO)[C@@H](O)[C@H](O)[C@H]5O[C@H]6O[C@H](CO)[C@@H](O)[C@H](O)[C@H]6O)C(C)(C)[C@@H]4CC[C@@]23C)C NJUXRKMKOFXMRX-AXUZFSSLSA-N 0.000 title claims abstract description 117
- NJUXRKMKOFXMRX-UHFFFAOYSA-N ginsenoside Rz1 Natural products CC(C)=CCC=C(C)C1CCC(C2(CCC3C4(C)C)C)(C)C1C(O)CC2C3(C)CCC4OC1OC(CO)C(O)C(O)C1OC1OC(CO)C(O)C(O)C1O NJUXRKMKOFXMRX-UHFFFAOYSA-N 0.000 title claims abstract description 117
- 238000000034 method Methods 0.000 title claims abstract description 37
- SYXUBXTYGFJFEH-UHFFFAOYSA-N oat triterpenoid saponin Chemical compound CNC1=CC=CC=C1C(=O)OC1C(C=O)(C)CC2C3(C(O3)CC3C4(CCC5C(C)(CO)C(OC6C(C(O)C(OC7C(C(O)C(O)C(CO)O7)O)CO6)OC6C(C(O)C(O)C(CO)O6)O)CCC53C)C)C4(C)CC(O)C2(C)C1 SYXUBXTYGFJFEH-UHFFFAOYSA-N 0.000 claims abstract description 61
- 239000011347 resin Substances 0.000 claims abstract description 58
- 229920005989 resin Polymers 0.000 claims abstract description 58
- 239000000178 monomer Substances 0.000 claims abstract description 55
- 239000002904 solvent Substances 0.000 claims abstract description 49
- 239000003463 adsorbent Substances 0.000 claims abstract description 40
- 238000004440 column chromatography Methods 0.000 claims abstract description 36
- 241000208343 Panax Species 0.000 claims abstract description 32
- 239000002994 raw material Substances 0.000 claims abstract description 29
- 239000000284 extract Substances 0.000 claims abstract description 25
- UFNDONGOJKNAES-UHFFFAOYSA-N Ginsenoside Rb1 Natural products CC(=CCCC(C)(OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CC(O)C45C)C UFNDONGOJKNAES-UHFFFAOYSA-N 0.000 claims abstract description 19
- GZYPWOGIYAIIPV-JBDTYSNRSA-N ginsenoside Rb1 Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)O[C@@](C)(CCC=C(C)C)[C@@H]1[C@@H]2[C@@]([C@@]3(CC[C@H]4C(C)(C)[C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)CC[C@]4(C)[C@H]3C[C@H]2O)C)(C)CC1)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GZYPWOGIYAIIPV-JBDTYSNRSA-N 0.000 claims abstract description 19
- TXEWRVNOAJOINC-UHFFFAOYSA-N ginsenoside Rb2 Natural products CC(=CCCC(OC1OC(COC2OCC(O)C(O)C2O)C(O)C(O)C1O)C3CCC4(C)C3C(O)CC5C6(C)CCC(OC7OC(CO)C(O)C(O)C7OC8OC(CO)C(O)C(O)C8O)C(C)(C)C6CCC45C)C TXEWRVNOAJOINC-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000010992 reflux Methods 0.000 claims abstract description 18
- 229930182494 ginsenoside Natural products 0.000 claims abstract description 10
- 229940089161 ginsenoside Drugs 0.000 claims abstract description 10
- 238000010025 steaming Methods 0.000 claims abstract description 8
- 239000003960 organic solvent Substances 0.000 claims description 325
- 238000010828 elution Methods 0.000 claims description 277
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 256
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 156
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 128
- 229930182490 saponin Natural products 0.000 claims description 59
- 150000007949 saponins Chemical class 0.000 claims description 59
- 239000000243 solution Substances 0.000 claims description 57
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 52
- 238000000605 extraction Methods 0.000 claims description 29
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 26
- 238000000746 purification Methods 0.000 claims description 24
- 239000002250 absorbent Substances 0.000 claims description 18
- 230000002745 absorbent Effects 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 13
- 238000001914 filtration Methods 0.000 claims description 6
- 239000012535 impurity Substances 0.000 claims description 6
- 238000011068 loading method Methods 0.000 claims description 6
- 238000012545 processing Methods 0.000 claims description 6
- 210000005036 nerve Anatomy 0.000 claims description 4
- 235000002791 Panax Nutrition 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 229930002875 chlorophyll Natural products 0.000 claims description 2
- 235000019804 chlorophyll Nutrition 0.000 claims description 2
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 claims description 2
- 241000252212 Danio rerio Species 0.000 abstract description 30
- 208000024827 Alzheimer disease Diseases 0.000 abstract description 9
- 206010039966 Senile dementia Diseases 0.000 abstract description 8
- 238000010521 absorption reaction Methods 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 2
- 239000011148 porous material Substances 0.000 abstract description 2
- 235000017709 saponins Nutrition 0.000 description 55
- 244000131316 Panax pseudoginseng Species 0.000 description 27
- 238000004128 high performance liquid chromatography Methods 0.000 description 27
- 235000003140 Panax quinquefolius Nutrition 0.000 description 23
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 22
- 235000008434 ginseng Nutrition 0.000 description 22
- 229930182470 glycoside Natural products 0.000 description 10
- 150000002338 glycosides Chemical class 0.000 description 10
- 239000000843 powder Substances 0.000 description 10
- 239000000344 soap Substances 0.000 description 10
- 241000180649 Panax notoginseng Species 0.000 description 8
- 235000003143 Panax notoginseng Nutrition 0.000 description 8
- ADEBPBSSDYVVLD-UHFFFAOYSA-N donepezil Chemical compound O=C1C=2C=C(OC)C(OC)=CC=2CC1CC(CC1)CCN1CC1=CC=CC=C1 ADEBPBSSDYVVLD-UHFFFAOYSA-N 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 7
- 206010012289 Dementia Diseases 0.000 description 5
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 239000000469 ethanolic extract Substances 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 239000007900 aqueous suspension Substances 0.000 description 4
- 229960003530 donepezil Drugs 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 230000037005 anaesthesia Effects 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012156 elution solvent Substances 0.000 description 3
- 239000000401 methanolic extract Substances 0.000 description 3
- 210000000653 nervous system Anatomy 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 208000012661 Dyskinesia Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000005238 degreasing Methods 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 230000007087 memory ability Effects 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 239000013558 reference substance Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 108090000312 Calcium Channels Proteins 0.000 description 1
- 102000003922 Calcium Channels Human genes 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 241000448255 Congiopodus peruvianus Species 0.000 description 1
- 241000283070 Equus zebra Species 0.000 description 1
- 102000001398 Granzyme Human genes 0.000 description 1
- 108060005986 Granzyme Proteins 0.000 description 1
- ZRJBHWIHUMBLCN-SEQYCRGISA-N Huperzine A Natural products N1C(=O)C=CC2=C1C[C@H]1/C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-SEQYCRGISA-N 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 102000001490 Opioid Peptides Human genes 0.000 description 1
- 108010093625 Opioid Peptides Proteins 0.000 description 1
- 240000005373 Panax quinquefolius Species 0.000 description 1
- ZRJBHWIHUMBLCN-UHFFFAOYSA-N Shuangyiping Natural products N1C(=O)C=CC2=C1CC1C(=CC)C2(N)CC(C)=C1 ZRJBHWIHUMBLCN-UHFFFAOYSA-N 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000002269 analeptic agent Substances 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000003542 behavioural effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 230000004715 cellular signal transduction Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000011208 chromatographic data Methods 0.000 description 1
- 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 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 1
- ZRJBHWIHUMBLCN-YQEJDHNASA-N huperzine A Chemical compound N1C(=O)C=CC2=C1C[C@H]1\C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-YQEJDHNASA-N 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000000366 juvenile effect Effects 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- HZVOZRGWRWCICA-UHFFFAOYSA-N methanediyl Chemical compound [CH2] HZVOZRGWRWCICA-UHFFFAOYSA-N 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 239000003399 opiate peptide Substances 0.000 description 1
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- ZRJBHWIHUMBLCN-BMIGLBTASA-N rac-huperzine A Natural products N1C(=O)C=CC2=C1C[C@@H]1C(=CC)[C@@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-BMIGLBTASA-N 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940072040 tricaine Drugs 0.000 description 1
- FQZJYWMRQDKBQN-UHFFFAOYSA-N tricaine methanesulfonate Chemical compound CS([O-])(=O)=O.CCOC(=O)C1=CC=CC([NH3+])=C1 FQZJYWMRQDKBQN-UHFFFAOYSA-N 0.000 description 1
- 150000008130 triterpenoid saponins Chemical class 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
- C07J17/005—Glycosides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
Abstract
The present invention relates to a kind of ginsenoside Rg 5 and ginsenoside Rk1 extracting method and application, belong to pharmaceutical technology field.Panax species raw material after the present invention will steam is extracted with solvent refluxing, extract solution obtains the total saposins containing ginsenoside Rg 5 and Rk1 through macroporous adsorbent resin column chromatography, total saposins, the general ginsenoside through steaming, arasaponin or ginsenoside Rb1 through steaming through macroporous adsorbent resin column chromatography are obtained into ginsenoside Rg 5 and Rk1 total contents be more than 97% group saponine, through large pore resin absorption column, column chromatography obtains the monomer ginsenoside Rg 5 and monomer ginsenoside Rk1 that purity is more than 90% to group saponine repeatedly again.Total saposins, group saponine and the monomer ginsenoside Rg 5 and monomer ginsenoside Rk1 that the present invention extracts can significantly improve zebra fish locomitivity and respond in alchlor induces senile dementia zebra fish model.
Description
Technical field
The present invention relates to a kind of ginsenoside Rg 5 and ginsenoside Rk1 extracting method and application, belongs to medical science neck
Domain.
Background technology
Pseudo-ginseng and the panax species such as ginseng, American Ginseng contain similar chemical composition, including saponin(e, flavones, carbene alcohol,
Polysaccharide etc., dammarane type four-ring triterpenoid saponin are one of major physiological active component of Panax medicinal plant.So far, it has been found that
Arasaponin and some monomeric compounds are in the side such as cardiovascular system, nervous system, metabolism and anti-inflammatory, antitumor
Face has preferable pharmacological activity, and the active reporter especially in terms of nervous system obtains more and more.Such as arasaponin and
Monomer saponin Rb1, Rg1 can significantly increase the learning and memory ability of mouse；Arasaponin has calm, analgesic activity, it
It is a kind of opioid peptides sample receptor stimulating agent, and it is not additive.General ginsenoside can be significantly improved with Alzheimer disease
The learning and memory ability of mouse, its therapeutic equivalence is in huperzine A；There is ginsenoside Rg 5 very strong NSC to activate
Effect, EGF reaction nerve stem cell directional can be induced to be divided into neuronal cell, and voltage door can be activated
The calcium channel of control, increase the calcium ion concentration in NSC.Ginsenoside Rg 5 generates available for Studies On Neuronal
During cellular signal transduction pathways, it is also possible to develop into treatment the nervous system disease new drug.But ginsenoside Rg 5
It is the micro constitutent in panax species, the yield that ginsenoside Rg 5 is isolated from panax species reported only has
0.0027%, this greatly limits ginsenoside Rg 5 to develop.Ginsenoside Rk1 be also in panax species it is micro into
Point, early-stage Study shows that ginsenoside Rk1 has and suppresses the bioactivity such as granzyme-induced Apoptosis and blocking vascular leakage.
But the report without ginsenoside Rk1 to nervous system effect.
The content of the invention
In view of the deficiencies of the prior art, the present invention provides the extracting method of a kind of ginsenoside Rg 5 and ginsenoside Rk1,
Ginsenoside Rg 5 and Rk1 content in panax species can be increased considerably and group saponine and monomer saponin can be obtained.
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）Panax species raw material is subjected to steam treatment, is then extracted to obtain extract solution with solvent refluxing, wherein solvent is second
Alcohol, methanol or water；
（2）By step（1）Solvent in gained extract solution volatilizees to obtain extract, and extract is dissolved in solvent B to obtain B molten
Liquid, filtering and impurity removing, then through macroporous adsorbent resin column chromatography, be eluted with water to sugar-free and distribute, then eluted with elution organic solvent C
Flow out to obtain the total saposins containing ginsenoside Rg 5 and Rk1 to without saponin(e, wherein solvent B be water, aqueous methanol, hydrous ethanol or
Aqueous acetone；
（3）General ginsenoside, arasaponin or ginsenoside Rb1 are subjected to steam treatment；
（4）By step（2）Gained total saposins or step（3）Gained, which steams product and is dissolved in, to be obtained total saposins in organic solvent D and has
Machine solution, total saposins organic solution are eluted, then washed with containing through macroporous adsorbent resin column chromatography with the E of organic solvent containing water elution
De- organic solvent F is eluted to no saponin(e and flows out to obtain the group saponine of ginsenoside Rg 5 and Rk1 total contents more than 97%, wherein having
Solvent D is ethanol or methanol；
（5）Purification：By step（4）Gained group saponine, which is dissolved in organic solvent H, obtains group saponine organic solution, its
Middle organic solvent H is ethanol, methanol, and group saponine organic solution is through macroporous adsorbent resin column chromatography, with organic molten containing water elution
Agent I elute, then respectively with concentration be 70 ~ 71% the J of organic solvent containing water elution, 71 ~ 72% organic solvent containing water elution K, 72 ~
73% L of organic solvent containing water elution, 73 ~ 74% organic solvent containing water elution M, 74 ~ 76% organic solvent containing water elution N enter
Row gradient elution flows out to without saponin(e, respectively obtains the flow point containing ginsenoside Rg 5 and Rk1, merges contain ginsenoside Rg 5 respectively
The flow point of ginsenoside Rg 5 and ginsenoside Rk1 flow points are obtained with Rk1 flow point；
（6）By step（5）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%.
The elution organic solvent is ethanol, methanol or acetone；
The step（1）The solid-to-liquid ratio kg of middle panax species raw material and solvent:L is 1:(8 ~ 10), the number of refluxing extraction is 3
~ 5 times, the time of each refluxing extraction is 2 ~ 4 h；
The step（2）The mass ratio of middle extract and macroporous absorbent resin is 1:(1.5 ~ 2.0), large pore resin absorption column layer
The loading flow velocity of B solution is 0.5 ~ 1 BV/h during analysis, during water elution the flow velocity of water be 2 ~ 3 BV/h, flow be 2 ~ 3 BV, elution
The flow velocity that organic solvent C is eluted when organic solvent C is eluted is 1 ~ 2 BV/h；
The step（4）Middle total saposins or the mass ratio for steaming product and macroporous absorbent resin are 1:(5 ~ 10), have containing water elution
Organic solvent E mass percent content is 65 ~ 70% in solvent E, organic solvent F matter in the F of organic solvent containing water elution
It is 70 ~ 75% to measure relative content, and the E of organic solvent containing water elution flow velocity is 1 ~ 3 BV/ when the E of organic solvent containing water elution is eluted
H, flow is 2 ~ 3 BV, and the F of organic solvent containing water elution flow velocity is 1 ~ 3 BV/h when the F of organic solvent containing water elution is eluted；
The step（5）The mass ratio of middle group saponine and macroporous absorbent resin is 1:(5 ~ 10), organic solvent I containing water elution
The mass percent content of middle organic solvent I is 70 ~ 75%, organic solvent I containing water elution when organic solvent I containing water elution elutes
Flow velocity be 2 ~ 3 BV/h, flow be 2 ~ 3 BV, the J of organic solvent containing water elution during gradient elution, the K of organic solvent containing water elution,
The L of organic solvent containing water elution, the M of organic solvent containing water elution, the N of organic solvent containing water elution flow velocity are 1 ~ 3 BV/h.
The panax species raw material is cauline leaf, step（2）Also include removing in the aqueous solution before macroporous adsorbent resin column chromatography
The processing procedure of chlorophyll；
The steaming method can be：Panax species raw material, general ginsenoside, arasaponin or ginsenoside Rb1 are put
Steamed in 120 ~ 135 DEG C of pressure cooker 12 ~ 15 hours；Or by panax species raw material, general ginsenoside, arasaponin
Or ginsenoside Rb1 is placed in 105 ~ 110 DEG C of reactor and steamed 10 ~ 12 hours；It is or panax species raw material, ginseng is total
The baking box that saponin(e, arasaponin or ginsenoside Rb1 are placed in 120 ~ 135 DEG C steams 12 ~ 15 hours；Or Panax is planted
Raw material, general ginsenoside, arasaponin or ginsenoside Rb1 are placed in 120 ~ 135 DEG C of autoclave that to steam 12 ~ 15 small
When；Or panax species raw material, general ginsenoside, arasaponin or ginsenoside Rb1 are placed in 100 ~ 125 DEG C of steaming
Steamed 18 ~ 24 hours in pot；
Water is added to soak panax species raw material, general ginsenoside, arasaponin or ginsenoside Rb1, then with gauze bag
Prick and carry out steam treatment, panax species are by steaming, and ginsenoside Rg 5 and Rk1 content can reach more than 4%, and ginseng is total
By steaming, ginsenoside Rg 5 and Rk1 content can reach more than 15% for saponin(e, arasaponin；Ginsenoside Rb1 passes through
Steam, ginsenoside Rg 5 and Rk1 content can reach more than 50%；
It is a further object of the present invention to provide what the extracting method of the ginsenoside Rg 5 and ginsenoside Rk1 were extracted to contain people
Total saposins, ginsenoside Rg 5 and the Rk1 total contents for joining saponin(e Rg5 and Rk1 are more than 97% group saponine or monomer ginsenoside
Rg5 and monomer ginsenoside Rk1 is improving the application of nerve retrograde affection.
Beneficial effects of the present invention：
（1）The inventive method can quickly dramatically increase content of the micro saponin(e in plant；
（2）The inventive method safety and environmental protection is rapidly and efficiently；
（3）The drug regimen that the present invention is prepared can effectively improve or alleviate symptom caused by senile dementia, available for old
The treatment of the nerve degenerative diseases such as dementia disease.
Brief description of the drawings
Fig. 1 is the structure chart of monomer saponin ginsenoside Rb1, Rg5 and Rk1；
Fig. 2 is the HPLC analysis result figures for the total saposins that embodiment 1 is extracted；
Fig. 3 is the HPLC analysis result figures for the group saponine that embodiment 1 is extracted；
Fig. 4 is the HPLC analysis result figures that the ginsenoside Rb1 of embodiment 3 cracks saponin(e；
Fig. 5 is the HPLC analysis result figures for the monomer saponin ginsenoside Rg 5 that embodiment 1 is extracted；
Fig. 6 is the HPLC analysis result figures for the monomer saponin ginsenoside Rk1 that embodiment 1 is extracted；
Fig. 7 be embodiment 1 extract ginsenoside Rk1 in 60 min to the influence figure of AD zebra fish movement velocitys, wherein #7
For ginsenoside Rk1, DPZ is positive controls（Donepezil）；
Fig. 8 is total saposins, group saponine monomer saponin ginsenoside Rg 5 and the Rk1 of the extraction of embodiment 1 to zebra fish movement velocity
Resume treatment efficiency chart；
Fig. 9 is total saposins, group saponine, monomer saponin ginsenoside Rg 5 and the Rk1 of the extraction of embodiment 1 to zebra fish speed difference
It is worth the therapeutic efficiency recovered.
Embodiment
The present invention is described in further detail with reference to embodiment, but protection scope of the present invention and unlimited
In the content.
HPLC analytic approach：
Embodiments of the invention select the high performance liquid chromatographs of Waters 2695/2996, Millennium32 chromatographic data pipes
Reason system；Reagent is from trifluoroacetic acid aqueous solution, ultra-pure water；Reference substance is voluntarily to isolate and purify；
Chromatographic condition：Agilent HC-C18（4.6 × 250 mm, 5 μm）Chromatographic column；The nm of Detection wavelength 203；25 DEG C of column temperature；Stream
It is mutually acetonitrile (A)-water (B) system gradient elution to move, the mL/min of flow velocity 1.Table 1 specific as follows,
Table 1
The preparation of reference substance solution：Precision weighs ginsenoside Rg respectively5The 5.19 mg and mg of ginsenoside Rk1 5.23, put
In 5 mL volumetric flasks, dissolved with methanol and be settled to graduation mark, shaken up.
The structure chart of monomer saponin ginsenoside Rb1, Rg5 and Rk1 is as shown in figure 1, monomer saponin ginseng soap as can be seen from Figure 1
Glycosides Rb1, Rg5 and Rk1 structure.
Embodiment 1：The present embodiment panax species raw material is beaten powder, is crossed 40 mesh sieves from dry Roots of Panax Notoginseng；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）By panax species raw material（Radix Notoginseng powder）Steam treatment is carried out, then uses solvent（Solvent is ethanol）Refluxing extraction obtains
To extract solution, wherein panax species raw material（Radix Notoginseng powder）With solvent（Ethanol）Solid-to-liquid ratio kg:L is 1:10, refluxing extraction
Number is 5 times, and the time of each refluxing extraction is 2 h；The method of steam treatment is that Radix Notoginseng powder is added into water-soaked, with gauze bag
Wrap up in, be placed in 120 DEG C of autoclave and steam 12 hours；
（2）By step（1）Solvent in gained extract solution（Ethanol）Volatilization obtains ethanol extract, and ethanol extract is dissolved in
Solvent B（Solvent B is hydrous ethanol）In obtain B solution, filtering and impurity removing, then through macroporous adsorbent resin column chromatography, be eluted with water to
Sugar-free distributes, then with elution organic solvent C（It is ethanol to elute organic solvent C）No saponin(e is eluted to flow out to obtain soap containing ginseng
Glycosides Rg5 and Rk1 total saposins, are analyzed through HPLC（See Fig. 2）, the quality percentage of ginsenoside Rg 5 in the total saposins of the present embodiment
Number content is 4.5%, and ginsenoside Rk1 mass percent content is 4.1%；Wherein ethanol extract and macroporous absorbent resin
Mass ratio is 1:1.5, the loading flow velocity of B solution is 0.5BV/h during macroporous adsorbent resin column chromatography, and the flow velocity of water is during water elution
2BV/h, flow 2BV, elution organic solvent C elutes organic solvent C flow velocity when eluting is 1BV/h；
（3）By step（2）Gained total saposins are dissolved in organic solvent D（Organic solvent D is ethanol）In to obtain total saposins organic molten
Liquid, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is second to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is ethanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC（See Fig. 3）, the matter of ginsenoside Rg 5 in group saponine
It is 56.7% to measure relative content, and ginsenoside Rk1 mass percent content is 44.1%；Wherein total saposins and macroporous absorption tree
The mass ratio of fat is 1:Organic solvent E in 10, the E of organic solvent containing water elution（Ethanol）Mass percent content be 65%, it is aqueous
Elute organic solvent F in organic solvent F（Ethanol）Mass percent content be 70%, the E of organic solvent containing water elution（Ethanol）Wash
The E of organic solvent containing water elution when de-（Ethanol）Flow velocity be 2BV/h, flow be 2 BV, the F of organic solvent containing water elution（Ethanol）Wash
The F of organic solvent containing water elution when de-（Ethanol）Flow velocity be 1BV/h；
（4）Purification：By step（3）Gained group saponine is dissolved in organic solvent H（Organic solvent H is ethanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
Again respectively with concentration be 71% the J of organic solvent containing water elution, 72% organic solvent containing water elution K, 73% containing water elution it is organic
Solvent L, 74% organic solvent containing water elution M, 75% organic solvent containing water elution N carry out gradient elution and flowed out to without saponin(e,
The flow point containing ginsenoside Rg 5 and Rk1 is respectively obtained, merges the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtains ginsenoside
Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is hydrous ethanol, group saponine and macroporous absorption
The mass ratio of resin is 1:10, organic solvent I in organic solvent I containing water elution（Ethanol）Mass percent content be 70%, contain
When water elution organic solvent I elutes the flow velocity of the organic solvent I containing water elution be 2BV/h, flow 2BV, it is aqueous during gradient elution
Elute organic solvent J, the K of organic solvent containing water elution, the L of organic solvent containing water elution, the M of organic solvent containing water elution, containing water elution
Organic solvent N flow velocity is 1BV/h；
（5）By step（4）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the monomer ginsenoside Rg 5 of the present embodiment purification（See Fig. 5）Content be 90.5 %, purifying carries
The monomer ginsenoside Rk1 taken（See Fig. 6）Content be 93.6 %.
Embodiment 2：The present embodiment is raw material from arasaponin；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）Arasaponin is subjected to steam treatment；The method of steam treatment is that arasaponin is added into aqueous suspension, is placed in 105
DEG C reactor in steam 10 hours；
（2）By step（1）Gained steams product and is dissolved in organic solvent D（Organic solvent D is ethanol）In to obtain total saposins organic
Solution, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is second to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is ethanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 55.3%, and ginsenoside Rk1 mass percent content is 42.7%；Wherein steam product and macroporous absorbent resin
Mass ratio is 1:Organic solvent E in 5, the E of organic solvent containing water elution（Ethanol）Mass percent content be 70 %, containing water elution
Organic solvent F in organic solvent F（Ethanol）Mass percent content be 71 %, the E of organic solvent containing water elution（Ethanol）Elution
When organic solvent containing water elution E（Ethanol）Flow velocity be 2.5 BV/h, flow be 2.5 BV, the F of organic solvent containing water elution（Second
Alcohol）The F of organic solvent containing water elution during elution（Ethanol）Flow velocity be 2 BV/h；
（3）Purification：By step（2）Gained group saponine is dissolved in organic solvent H（Organic solvent H is ethanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
Contain water elution with the J of organic solvent containing water elution that concentration is 70%, the 71 % K of organic solvent containing water elution, 72% respectively again
Organic solvent L, the 73 % M of organic solvent containing water elution, 74% organic solvent containing water elution N carry out gradient elution to without saponin(e
Outflow, respectively obtains the flow point containing ginsenoside Rg 5 and Rk1, merges the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtain people
Join saponin(e Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is hydrous ethanol, group saponine with it is big
The mass ratio of macroporous adsorbent resin is 1:8, organic solvent I in organic solvent I containing water elution（Ethanol）Mass percent content be
70%, organic solvent I containing water elution elute when the organic solvent I containing water elution flow velocity be 2.5 BV/h, flow be 2.5 BV, ladder
It is organic solvent containing water elution J during degree elution, the K of organic solvent containing water elution, the L of organic solvent containing water elution, organic molten containing water elution
Agent M, the N of organic solvent containing water elution flow velocity are 2.5 BV/h；
（4）By step（3）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the content of the monomer ginsenoside Rg 5 of the present embodiment purification is 90.4%, the monomer of purification
Ginsenoside Rk1 content is 94.1%.
Embodiment 3：The present embodiment is raw material from ginsenoside Rb1；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）Ginsenoside Rb1 is subjected to steam treatment；The method of steam treatment is that arasaponin is added into aqueous suspension, is placed in 105
DEG C reactor in steam 10 hours；
（2）By step（1）Gained steams product and is dissolved in organic solvent D（Organic solvent D is ethanol）In to obtain total saposins organic
Solution, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is second to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is ethanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 50.4%, and ginsenoside Rk1 mass percent content is 47.9%；Wherein steam product and macroporous absorbent resin
Mass ratio is 1:Organic solvent E in 5, the E of organic solvent containing water elution（Ethanol）Mass percent content be 70 %, containing water elution
Organic solvent F in organic solvent F（Ethanol）Mass percent content be 71 %, the E of organic solvent containing water elution（Ethanol）Elution
When organic solvent containing water elution E（Ethanol）Flow velocity be 2.5 BV/h, flow be 2.5 BV, the F of organic solvent containing water elution（Second
Alcohol）The F of organic solvent containing water elution during elution（Ethanol）Flow velocity be 2 BV/h；
（3）By step（2）Gained group saponine is dissolved in organic solvent H（Organic solvent H is hydrous ethanol）In, through macroporous absorption
Resin column chromatography, eluted with organic solvent I containing water elution, then respectively with the J of organic solvent containing water elution that concentration is 70 %, 71%
The K of organic solvent containing water elution, the 72 % L of organic solvent containing water elution, 73% organic solvent containing water elution M, 75% it is aqueous
Elute organic solvent N and carry out gradient elution to without saponin(e outflow, respectively obtain the flow point containing ginsenoside Rg 5 and Rk1, close respectively
And the flow point containing ginsenoside Rg 5 and Rk1 obtains the flow point of ginsenoside Rg 5 and ginsenoside Rk1 flow points；Wherein have containing water elution
Solvent is hydrous ethanol, and the mass ratio of group saponine and macroporous absorbent resin is 1:10, in organic solvent I containing water elution
Organic solvent I（Ethanol）Mass percent content be 70 %, organic solvent containing water elution when organic solvent I containing water elution elutes
I flow velocity is 3BV/h, flow 3BV, the J of organic solvent containing water elution during gradient elution, the K of organic solvent containing water elution, aqueous
It is 3 BV/h to elute organic solvent L, the M of organic solvent containing water elution, the N of organic solvent containing water elution flow velocity；
（4）By step（3）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
The present embodiment ginsenoside Rb1 cracks the HPLC analysis results figure of saponin(e as shown in figure 4, being analyzed through HPLC, the present embodiment
The content of the monomer ginsenoside Rg 5 of purification is 95.1%, and the monomer ginsenoside Rk1 of purification content is
94.1%。
Embodiment 4：The present embodiment panax species raw material is beaten powder, is crossed 40 mesh sieves from dry ginseng；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）By panax species raw material（Ginseng pulverate）Steam treatment is carried out, then uses solvent（Solvent is ethanol）Refluxing extraction obtains
To extract solution, wherein panax species raw material（Ginseng pulverate）With solvent（Ethanol）Solid-to-liquid ratio kg:L is 1:8, time of refluxing extraction
Number is 3, and the time of each refluxing extraction is 4 h；The method of steam treatment is that ginseng pulverate is added into water-soaked, is placed in 120 DEG C of height
Steamed 15 hours in pressure pot；
（2）By step（1）Solvent in gained extract solution（Ethanol）Volatilization obtains ethanol extract, is mixed in ethanol extract plus water
Close and uniformly obtain suspension, then use petroleum ether extraction suspension respectively 5 times, degreasing, evaporation removes water and obtains extract, will extract
Thing is dissolved in solvent B（Solvent B is aqueous methanol）In obtain B solution, filtering and impurity removing, then through macroporous adsorbent resin column chromatography, use
Water elution to sugar-free distributes, then with elution organic solvent C（It is ethanol to elute organic solvent C）No saponin(e is eluted to flow out to obtain
Total saposins containing ginsenoside Rg 5 and Rk1, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in the total saposins of the present embodiment
Number content is 4.3%, and ginsenoside Rk1 mass percent content is 4.0%；The wherein quality of extract and macroporous absorbent resin
Than for 1:2, the loading flow velocity of B solution is 0.6BV/h during macroporous adsorbent resin column chromatography, and the flow velocity of water is 3BV/ during water elution
H, flow is 2 BV, and elution organic solvent C elutes organic solvent C flow velocity when eluting is 1.5 BV/h；
（3）By step（2）Gained total saposins are dissolved in organic solvent D（Organic solvent D is ethanol）In to obtain total saposins organic molten
Liquid, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is second to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is ethanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 50.2%, and ginsenoside Rk1 mass percent content is 47.6%；The wherein matter of total saposins and macroporous absorbent resin
Amount is than being 1:Organic solvent E in 9, the E of organic solvent containing water elution（Ethanol）Mass percent content be 70%, containing water elution
Organic solvent F in organic solvent F（Ethanol）Mass percent content be 75%, the E of organic solvent containing water elution（Ethanol）Elution
When organic solvent containing water elution E（Ethanol）Flow velocity be 3.0BV/h, flow be 2.5 BV, the F of organic solvent containing water elution（Ethanol）
The F of organic solvent containing water elution during elution（Ethanol）Flow velocity be 1.8 BV/h；
（4）Purification：By step（3）Gained group saponine is dissolved in organic solvent H（Organic solvent H is ethanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
The J of organic solvent containing water elution, 71% organic solvent containing water elution K, 72 % for being respectively again 70% with concentration have containing water elution
Solvent L, 73% organic solvent containing water elution M, the 75 % N of organic solvent containing water elution carry out gradient elution to without saponin(e stream
Go out, respectively obtain the flow point containing ginsenoside Rg 5 and Rk1, merge the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtain ginseng
Saponin(e Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is hydrous ethanol, group saponine and macropore
The mass ratio of polymeric adsorbent is 1:5, organic solvent I in organic solvent I containing water elution（Ethanol）Mass percent content be 75
%, when organic solvent I containing water elution elutes the flow velocity of the organic solvent I containing water elution be 3 BV/h, flow be 3 BV, gradient elution
When organic solvent containing water elution J, the K of organic solvent containing water elution, the L of organic solvent containing water elution, the M of organic solvent containing water elution, contain
Water elution organic solvent N flow velocity is 1.5 BV/h；
（5）By step（4）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the content of the monomer ginsenoside Rg 5 of the present embodiment purification is 90.4%, the monomer of purification
Ginsenoside Rk1 content is 93.9%.
Embodiment 5：The present embodiment panax species raw material is beaten powder, is crossed 40 mesh sieves from dry Roots of Panax Notoginseng；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）By panax species raw material（Radix Notoginseng powder）Steam treatment is carried out, then uses solvent（Solvent is methanol）Refluxing extraction obtains
To extract solution, wherein panax species raw material（Radix Notoginseng powder）With solvent（Methanol）Solid-to-liquid ratio kg:L is 1:9, refluxing extraction
Number is 4, and the time of each refluxing extraction is 3 h；The method of steam treatment is that Radix Notoginseng powder is added into water-soaked, with gauze wrapped,
It is placed in 105 DEG C of reactor and steams 10 hours；
（2）By step（1）Solvent in gained extract solution（Methanol）Volatilization obtains methanolic extract, and methanolic extract is dissolved in
Solvent B（Solvent B is water）In obtain B solution, filtering and impurity removing, then through macroporous adsorbent resin column chromatography, be eluted with water to without sugar
Outflow, then with elute organic solvent C（It is methanol to elute organic solvent C）It is eluted to no saponin(e and flows out to obtain and contains ginsenoside Rg 5
With Rk1 total saposins, to be analyzed through HPLC, the mass percent content of ginsenoside Rg 5 is 5.2% in the total saposins of the present embodiment,
Ginsenoside Rk1 mass percent content is 4.3%；Wherein the mass ratio of methanolic extract and macroporous absorbent resin is 1:2.0
, the loading flow velocity of B solution is 1.0BV/h during macroporous adsorbent resin column chromatography, during water elution the flow velocity of water be 2.8 BV/h, stream
Measure as 2.7 BV, the flow velocity that organic solvent C is eluted during the C elutions of elution organic solvent is 1.6 BV/h；
（3）By step（2）Gained total saposins are dissolved in organic solvent D（Organic solvent D is methanol）In to obtain total saposins organic molten
Liquid, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is first to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is methanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 50.9%, and ginsenoside Rk1 mass percent content is 46.5%；The wherein matter of total saposins and macroporous absorbent resin
Amount is than being 1:Organic solvent E in 5, the E of organic solvent containing water elution（Methanol）Mass percent content be 65%, containing water elution
Organic solvent F in organic solvent F（Methanol）Mass percent content be 70%, the E of organic solvent containing water elution（Methanol）During elution
The E of organic solvent containing water elution（Methanol）Flow velocity be 2 BV/h, flow be 3.0 BV, the F of organic solvent containing water elution（Methanol）Wash
The F of organic solvent containing water elution when de-（Methanol）Flow velocity be 3 BV/h；
（4）Purification：By step（3）Gained group saponine is dissolved in organic solvent H（Organic solvent H is methanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
Contain water elution with the J of organic solvent containing water elution that concentration is 70%, the 71 % K of organic solvent containing water elution, 72% respectively again
Organic solvent L, 74% organic solvent containing water elution M, 75% organic solvent containing water elution N carry out gradient elution to without saponin(e stream
Go out, respectively obtain the flow point containing ginsenoside Rg 5 and Rk1, merge the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtain ginseng
Saponin(e Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is aqueous methanol, group saponine and macropore
The mass ratio of polymeric adsorbent is 1:8, organic solvent I in organic solvent I containing water elution（Methanol）Mass percent content be
75%, when organic solvent I containing water elution elutes the flow velocity of the organic solvent I containing water elution be 3BV/h, flow be 3 BV, gradient elution
When organic solvent containing water elution J, the K of organic solvent containing water elution, the L of organic solvent containing water elution, the M of organic solvent containing water elution, contain
Water elution organic solvent N flow velocity is 3BV/h；
（5）By step（4）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the content of the monomer ginsenoside Rg 5 of the present embodiment purification is 90.5%, the monomer of purification
Ginsenoside Rk1 content is 93.8%.
Embodiment 6：The present embodiment is raw material from arasaponin；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）Arasaponin is subjected to steam treatment；The method of steam treatment is that arasaponin is added into aqueous suspension, is placed in 120
DEG C high-pressure sterilizing pot in steam 12 hours；
（2）By step（1）Gained steams product and is dissolved in organic solvent D（Organic solvent D is methanol）In to obtain total saposins organic
Solution, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is first to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is methanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 52.7%, and ginsenoside Rk1 mass percent content is 44.6%；The wherein matter of total saposins and macroporous absorbent resin
Amount is than being 1:Organic solvent E in 7, the E of organic solvent containing water elution（Methanol）Mass percent content be 68%, have containing water elution
Organic solvent F in solvent F（Methanol）Mass percent content be 72%, the E of organic solvent containing water elution（Methanol）Contain during elution
Water elution organic solvent E（Methanol）Flow velocity be 1.9 BV/h, flow be 2.8 BV, the F of organic solvent containing water elution（Methanol）Wash
The F of organic solvent containing water elution when de-（Methanol）Flow velocity be 2.6 BV/h；
（3）Purification：By step（2）Gained group saponine is dissolved in organic solvent H（Organic solvent H is methanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
The J of organic solvent containing water elution, 72% organic solvent containing water elution K, 73 % for being respectively again 70% with concentration have containing water elution
Solvent L, 74% organic solvent containing water elution M, 75% organic solvent containing water elution N carry out gradient elution to without saponin(e stream
Go out, respectively obtain the flow point containing ginsenoside Rg 5 and Rk1, merge the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtain ginseng
Saponin(e Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is aqueous methanol, group saponine and macropore
The mass ratio of polymeric adsorbent is 1:6, organic solvent I in organic solvent I containing water elution（Methanol）Mass percent content be
72%, when organic solvent I containing water elution elutes the flow velocity of the organic solvent I containing water elution be 3 BV/h, flow 3BV, gradient washes
The J of organic solvent containing water elution when de-, the K of organic solvent containing water elution, the L of organic solvent containing water elution, the M of organic solvent containing water elution,
The N of organic solvent containing water elution flow velocity is 3BV/h；
（4）By step（3）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the content of the monomer ginsenoside Rg 5 of the present embodiment purification is 91.4%, the monomer of purification
Ginsenoside Rk1 content is 94.3%.
Embodiment 7：The present embodiment is raw material from ginsenoside Rb1；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）Ginsenoside Rb1 is subjected to steam treatment；The method of steam treatment is that ginsenoside Rb1 is added into aqueous suspension, is placed in
Steamed in 120 DEG C of high-pressure sterilizing pot 15 hours；
（2）By step（1）Gained steams product and is dissolved in organic solvent D（Organic solvent D is methanol）In to obtain total saposins organic
Solution, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is first to elute organic solvent E
Alcohol）Elution, then with the F of organic solvent containing water elution（It is methanol to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 50.6%, and ginsenoside Rk1 mass percent content is 47.8%；The wherein matter of total saposins and macroporous absorbent resin
Amount is than being 1:Organic solvent E in 8, the E of organic solvent containing water elution（Methanol）Mass percent content be 69%, containing water elution
Organic solvent F in organic solvent F（Methanol）Mass percent content be 70 %, the E of organic solvent containing water elution（Methanol）Elution
When organic solvent containing water elution E（Methanol）Flow velocity be 2 BV/h, flow be 2 BV, the F of organic solvent containing water elution（Methanol）Wash
The F of organic solvent containing water elution when de-（Methanol）Flow velocity be 2BV/h；
（3）Purification：By step（2）Gained group saponine is dissolved in organic solvent H（Organic solvent H is methanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
That uses the J of organic solvent containing water elution, 72% organic solvent containing water elution K, 73% that concentration is 71% respectively again contains water elution
Organic solvent L, 74% organic solvent containing water elution M, 76% organic solvent containing water elution N carry out gradient elution to without saponin(e
Outflow, respectively obtains the flow point containing ginsenoside Rg 5 and Rk1, merges the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtain people
Join saponin(e Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is aqueous methanol, group saponine with it is big
The mass ratio of macroporous adsorbent resin is 1:10, organic solvent I in organic solvent I containing water elution（Methanol）Percentage by volume content
For 70%, when organic solvent I containing water elution elutes the flow velocity of the organic solvent I containing water elution be 3BV/h, flow 2BV, gradient washes
The J of organic solvent containing water elution when de-, the K of organic solvent containing water elution, the L of organic solvent containing water elution, the M of organic solvent containing water elution,
The N of organic solvent containing water elution flow velocity is 1BV/h；
（4）By step（3）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the content of the monomer ginsenoside Rg 5 of the present embodiment purification is 95.3%, the monomer of purification
Ginsenoside Rk1 content is 94.4%.
Embodiment 8：The present embodiment panax species raw material is beaten powder, is crossed 40 mesh sieves from dry pseudo-ginseng fibrous root；
A kind of extracting method of ginsenoside Rg 5 and ginsenoside Rk1, is comprised the following steps that：
（1）By panax species raw material（Pseudo-ginseng fibrous root）Steam treatment is carried out, then uses solvent（Solvent is water）Refluxing extraction obtains
To extract solution, wherein panax species raw material（Pseudo-ginseng fibrous root）With solvent（Water）Solid-to-liquid ratio kg:L is 1:9, refluxing extraction
Number is 4, and the time of each refluxing extraction is 3h；The method of steam treatment is that pseudo-ginseng fibrous root point is added into water-soaked, with gauze bag
Wrap up in and be placed in closed vessel, be placed in baking box and steam 15 hours；
（2）By step（1）Solvent in gained extract solution（Water）Volatilization obtains water extract, is mixed with water in water extract
Suspension is obtained, then uses petroleum ether extraction suspension respectively 5 times, degreasing, evaporation removes water and obtains extract, and extract is dissolved
In solvent B（Solvent B is aqueous acetone）In obtain B solution, filtering and impurity removing, then through macroporous adsorbent resin column chromatography, be eluted with water
Distributed to sugar-free, then with elution organic solvent C（It is acetone to elute organic solvent C）It is eluted to no saponin(e and flows out to obtain and contains ginseng
Saponin(e Rg5 and Rk1 total saposins, are analyzed through HPLC, the mass percent content of ginsenoside Rg 5 in the total saposins of the present embodiment
For 4.1%, ginsenoside Rk1 mass percent content is 4.0%；Wherein the mass ratio of extract and macroporous absorbent resin is 1:
1.8, the loading flow velocity of B solution is 0.8 BV/h during macroporous adsorbent resin column chromatography, during water elution the flow velocity of water be 2.5 BV/h,
Flow is 2.5 BV, and elution organic solvent C elutes organic solvent C flow velocity when eluting is 2.0 BV/h；
（3）By step（2）Gained total saposins are dissolved in organic solvent D（Organic solvent D is methanol）In to obtain total saposins organic molten
Liquid, total saposins organic solution is through macroporous adsorbent resin column chromatography, with the E of organic solvent containing water elution（It is third to elute organic solvent E
Ketone）Elution, then with the F of organic solvent containing water elution（It is acetone to elute organic solvent F）No saponin(e is eluted to flow out to obtain ginseng soap
Glycosides Rg5 and Rk1 total content are more than 97% group saponine, are analyzed through HPLC, the quality percentage of ginsenoside Rg 5 in group saponine
Number content is 50.2%, and ginsenoside Rk1 mass percent content is 47.6%；The wherein matter of total saposins and macroporous absorbent resin
Amount is than being 1:Organic solvent E in 10, the E of organic solvent containing water elution（Acetone）Percentage by volume content be 68%, have containing water elution
Organic solvent F in solvent F（Acetone）Percentage by volume content be 70%, the E of organic solvent containing water elution（Acetone）Contain during elution
Water elution organic solvent E（Acetone）Flow velocity be 1BV/h, flow 2BV, the F of organic solvent containing water elution（Acetone）Contain during elution
Water elution organic solvent F（Acetone）Flow velocity be 3 BV/h；
（4）Purification：By step（3）Gained group saponine is dissolved in organic solvent H（Organic solvent H is methanol）In divided
Saponins organic solution, group saponine organic solution are eluted with organic solvent I containing water elution through macroporous adsorbent resin column chromatography,
Have respectively with the J of organic solvent containing water elution that concentration is 70%, 71% organic solvent containing water elution K, 72% containing water elution again
Solvent L, 73% organic solvent containing water elution M, 75% organic solvent containing water elution N carry out gradient elution to without saponin(e stream
Go out, respectively obtain the flow point containing ginsenoside Rg 5 and Rk1, merge the flow point containing ginsenoside Rg 5 and Rk1 respectively and obtain ginseng
Saponin(e Rg5 flow points and ginsenoside Rk1 flow points；Wherein organic solvent containing water elution is aqueous methanol, group saponine and macropore
The mass ratio of polymeric adsorbent is 1:10, organic solvent I in organic solvent I containing water elution（Methanol）Mass percent content be
70%, when organic solvent I containing water elution elutes the flow velocity of the organic solvent I containing water elution be 2BV/h, flow 2BV, gradient washes
The J of organic solvent containing water elution when de-, the K of organic solvent containing water elution, the L of organic solvent containing water elution, the M of organic solvent containing water elution,
The N of organic solvent containing water elution flow velocity is 2BV/h；
（5）By step（4）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%；
Analyzed through HPLC, the content of the monomer ginsenoside Rg 5 of the present embodiment purification is 92.7%, the monomer of purification
Ginsenoside Rk1 content is 95.1%.
Embodiment 9：Total saposins, group saponine, ginsenoside Rg 5 and the list that the present embodiment is prepared using embodiment 1
Body ginsenoside Rk1 activity
The experimental animal that the present embodiment uses for zebra fish, zebra fish juvenile fish by zebra fingerling fish produce embryo be incubated naturally and
Into；Fish culture water water quality：200 mg Instant Oceans are added in per 1L reverse osmosis waters, electrical conductivity is 480 ~ 510 μ S/cm；PH is
6.9~7.2；Hardness is 53.7 ~ 71.6 mg/L CaCO3；After the completion of experiment, with 0.25mg/ml tricaine methanesulfonic acid pair
The zebra fish of each stage of development carries out anesthesia execution.The operating procedure that anesthesia is put to death meets American Veterinary association（AVMA）To dynamic
The code requirement that thing anesthesia is put to death；
The present embodiment sets 5 groups of experiments altogether, and every group of experiment includes 1 blank control group, 1 solvent control group, 1 senile dementia
Model group, 1 positive controls（Donepezil）, 3-5 testing compound group；The primary dcreening operation concentration of testing compound is 30 μM,
If there is dead or deformity, choose 10 μM and tested；The zebra fish of alchlor processing is Senlie dementia model group；Three
Aluminium chloride is positive controls with the DPZ zebra fish being jointly processed by；The zebra fish that alchlor is jointly processed by with testing compound
For testing compound group；The zebra fish of 0.1% DMSO processing is negative control group（Solvent control group）；Untreated zebra fish
For blank control group, each experimental group handles 30 tail zebra fish；The fortune of zebra fish in 60 min is recorded using behavioural analysis instrument
Dynamic rail mark, is changed by light dark period, 10 min of dark, and illumination 10 min, 60 min point are three light dark periods, then right
The movement velocity of zebra fish（Dyskinesia）, light and shade period velocity change (respond）Carry out statistical analysis；
Active testing result meets quality control standard：1. all experimental groups do not occur zebra fish death in whole experiment process
Or deformity；2. model group and solvent control group compare with significant difference（p < 0.001）;3. blank pair in all experiments
According to group compared with solvent control group no difference of science of statistics（p > 0.05）；4. positive controls（Donepezil, DPZ）With model
Group compares with significant difference（p < 0.05）；
To the restitution of senile dementia zebra fish dyskinesia（See Fig. 8 and table 2）
The therapeutic efficiency of the senile dementia zebra fish movement velocity of table 2
Influence figures of the ginsenoside Rk1 of the present embodiment in 60 min to AD zebra fish movement velocitys is as shown in fig. 7, wherein #
7 be that ginsenoside Rk1, DPZ are positive controls（Donepezil）, as can be known from Fig. 7, ginsenoside Rk1 can effectively improve three
The movement velocity of the senile dementia zebra fish of aluminum oxide induction；
It was found from Fig. 8 and table 2, all experiment Senlie dementia model group zebra fish movement velocitys are remarkably decreased, with solvent control group
Compare statistically significant (p< 0.001)；Positive drug (DPZ) group zebra fish movement velocity is significantly gone up, with model group ratio
Relatively there is statistical significance (p<0.01 or p< 0.001).Within the 60min light and shade cycles, its movement velocity has zebra fish
Regular, periodicity and stability；AD spots after total saposins, group saponine and monomer saponin processing that embodiment 1 is prepared
The movement velocity of horse fish significantly improves (p compared with model group< 0.05, p <0.01 or p<0.001), significantly recover old
The movement velocity of dementia disease zebra fish；
To the restitution of senile dementia zebra fish respond（See Fig. 9 and table 3）
The therapeutic efficiency of the senile dementia zebra fish respond of table 3
It was found from Fig. 9 and table 3, in all experiments, Senlie dementia model group zebra fish respond declines（Light and shade period velocity is poor
Value is remarkably decreased）, with the more statistically significant (p of solvent control group< 0.001)；Positive drug (DPZ) group zebra fish reaction
Part ability recovers（Light and shade period velocity difference is significantly gone up）, with statistical significance (p compared with model group<0.05 or p
< 0.001).Recovery AD that can be in various degree after total saposins that embodiment 1 is prepared, group saponine and monomer saponin processing
The reaction speed of zebra fish（Light and shade period velocity difference is significantly gone up）, compared with model group, significant difference is significantly or extremely aobvious
Write (p< 0.05, p <0.01 or p< 0.001).
Claims (8)
1. a kind of ginsenoside Rg 5 and ginsenoside Rk1 extracting method, it is characterised in that comprise the following steps that：
（1）Panax species raw material is subjected to steam treatment, is then extracted to obtain extract solution with solvent refluxing, wherein solvent is second
Alcohol, methanol or water；
（2）By step（1）Solvent in gained extract solution volatilizees to obtain extract, and extract is dissolved in solvent B to obtain B molten
Liquid, filtering and impurity removing, then through macroporous adsorbent resin column chromatography, be eluted with water to sugar-free and distribute, then eluted with elution organic solvent C
Flow out to obtain the total saposins containing ginsenoside Rg 5 and Rk1 to without saponin(e, wherein solvent B be water, aqueous methanol, hydrous ethanol or
Aqueous acetone；
（3）General ginsenoside, arasaponin or ginsenoside Rb1 are subjected to steam treatment；
（4）By step（2）Gained total saposins or step（3）Gained, which steams product and is dissolved in, to be obtained total saposins in organic solvent D and has
Machine solution, total saposins organic solution are eluted, then washed with containing through macroporous adsorbent resin column chromatography with the E of organic solvent containing water elution
De- organic solvent F is eluted to no saponin(e and flows out to obtain the group saponine of ginsenoside Rg 5 and Rk1 total contents more than 97%, wherein having
Solvent D is ethanol or methanol；
（5）Purification：By step（4）Gained group saponine, which is dissolved in organic solvent H, obtains group saponine organic solution, its
Middle organic solvent H is ethanol, methanol, and group saponine organic solution is through macroporous adsorbent resin column chromatography, with organic molten containing water elution
Agent I elute, then respectively with concentration be 70 ~ 71% the J of organic solvent containing water elution, 71 ~ 72% organic solvent containing water elution K, 72 ~
73% L of organic solvent containing water elution, 73 ~ 74% organic solvent containing water elution M, 74 ~ 76% organic solvent containing water elution N enter
Row gradient elution flows out to without saponin(e, respectively obtains the flow point containing ginsenoside Rg 5 and Rk1, merges contain ginsenoside Rg 5 respectively
The flow point of ginsenoside Rg 5 and ginsenoside Rk1 flow points are obtained with Rk1 flow point；
（6）By step（5）Gained ginsenoside Rg5 flow points and ginsenoside Rk1 flow points repeat step respectively（4）Purifying
Extraction process obtains the monomer ginsenoside Rk1 of monomer ginsenoside Rg 5 of the content more than 90% and content more than 90%.
2. ginsenoside Rg 5 and ginsenoside Rk1 extracting method according to claim 1, it is characterised in that：Elute organic
Solvent is ethanol, methanol or acetone.
3. ginsenoside Rg 5 and ginsenoside Rk1 extracting method according to claim 1, it is characterised in that：Step（1）
The solid-to-liquid ratio kg of middle panax species raw material and solvent:L is 1:(8 ~ 10), the number of refluxing extraction is 3 ~ 5 times, and backflow every time carries
The time taken is 2 ~ 4 h.
4. ginsenoside Rg 5 and ginsenoside Rk1 extracting method according to claim 1, it is characterised in that：Step（2）
The mass ratio of middle extract and macroporous absorbent resin is 1:(1.5 ~ 2.0), the loading of B solution during macroporous adsorbent resin column chromatography
Flow velocity is 0.5 ~ 1BV/h, during water elution the flow velocity of water be 2 ~ 3BV/h, flow be 2 ~ 3BV, elute during the C elutions of elution organic solvent
Organic solvent C flow velocity is 1 ~ 2BV/h.
5. ginsenoside Rg 5 and ginsenoside Rk1 extracting method according to claim 1, it is characterised in that：Step（4）
Middle total saposins or the mass ratio for steaming product and macroporous absorbent resin are 1:(5 ~ 10), it is organic molten in the E of organic solvent containing water elution
Agent E mass percent content is 65 ~ 70%, and organic solvent F mass percent content is in the F of organic solvent containing water elution
When 70 ~ 75%, the E of organic solvent containing water elution are eluted organic solvent containing water elution E flow velocity be 1 ~ 3BV/h, flow be 2 ~ 3 BV,
The F of organic solvent containing water elution flow velocity is 1 ~ 3 BV/h when the F of organic solvent containing water elution is eluted.
6. ginsenoside Rg 5 and ginsenoside Rk1 extracting method according to claim 1, it is characterised in that：Step（5）
The mass ratio of middle group saponine and macroporous absorbent resin is 1:(5 ~ 10), the matter of organic solvent I in organic solvent I containing water elution
It is 70 ~ 75% to measure relative content, and the flow velocity of the organic solvent I containing water elution is 2 ~ 3 BV/ when organic solvent I containing water elution elutes
H, flow is 2 ~ 3 BV, the J of organic solvent containing water elution, the K of organic solvent containing water elution, organic solvent containing water elution during gradient elution
L, the M of organic solvent containing water elution, the N of organic solvent containing water elution flow velocity are 1 ~ 3 BV/h.
7. ginsenoside Rg 5 and ginsenoside Rk1 extracting method according to claim 1, it is characterised in that：Panax is planted
Raw material is cauline leaf, step（2）Also include the processing procedure except aqueous solution Determination of Chlorophyll before macroporous adsorbent resin column chromatography.
8. the He containing ginsenoside Rg 5 that ginsenoside Rg 5 and ginsenoside Rk1 extracting method described in claim 1 are extracted
Group saponine of Rk1 total saposins, ginsenoside Rg 5 and the Rk1 total contents more than 90% or monomer ginsenoside Rg 5 and monomer people
Joining saponin(e Rk1 is improving the application of nerve retrograde affection.
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| CN111087438A (en) * | 2020-01-04 | 2020-05-01 | 丁传波 | Preparation method of rare saponins of panax ginseng Rk1 and Rg5 |
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| CN103030678A (en) * | 2012-12-24 | 2013-04-10 | 吉林农业大学 | Method for preparing rare ginsenoside from malonyl ginsenoside |
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| CN103030678A (en) * | 2012-12-24 | 2013-04-10 | 吉林农业大学 | Method for preparing rare ginsenoside from malonyl ginsenoside |
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| DONG WANG ET AL.: "The processing of Panax notoginseng and the transformation of its saponin components", 《FOOD CHEMISTRY》 * |
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