CN116925163A - Method for obtaining rose glycoside by low-frequency ultrasound - Google Patents
Method for obtaining rose glycoside by low-frequency ultrasound Download PDFInfo
- Publication number
- CN116925163A CN116925163A CN202310833880.XA CN202310833880A CN116925163A CN 116925163 A CN116925163 A CN 116925163A CN 202310833880 A CN202310833880 A CN 202310833880A CN 116925163 A CN116925163 A CN 116925163A
- Authority
- CN
- China
- Prior art keywords
- rose
- ultrasonic
- aqueous solution
- reduced pressure
- glycoside
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000220317 Rosa Species 0.000 title claims abstract description 88
- 229930182470 glycoside Natural products 0.000 title claims abstract description 59
- 150000002338 glycosides Chemical class 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 31
- 238000002604 ultrasonography Methods 0.000 title claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 87
- 239000000243 solution Substances 0.000 claims abstract description 67
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 46
- 239000002202 Polyethylene glycol Substances 0.000 claims abstract description 34
- 229920001223 polyethylene glycol Polymers 0.000 claims abstract description 34
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 33
- 238000000605 extraction Methods 0.000 claims abstract description 29
- 239000011259 mixed solution Substances 0.000 claims abstract description 27
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 23
- 239000011780 sodium chloride Substances 0.000 claims abstract description 23
- 239000002994 raw material Substances 0.000 claims abstract description 20
- -1 polyethylene Polymers 0.000 claims abstract description 12
- 238000007670 refining Methods 0.000 claims abstract description 11
- 239000004698 Polyethylene Substances 0.000 claims abstract description 10
- 229920000573 polyethylene Polymers 0.000 claims abstract description 10
- FWFGVMYFCODZRD-UHFFFAOYSA-N oxidanium;hydrogen sulfate Chemical compound O.OS(O)(=O)=O FWFGVMYFCODZRD-UHFFFAOYSA-N 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 46
- 239000007864 aqueous solution Substances 0.000 claims description 42
- 238000001704 evaporation Methods 0.000 claims description 38
- 230000008020 evaporation Effects 0.000 claims description 38
- 239000006228 supernatant Substances 0.000 claims description 37
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 36
- 238000002156 mixing Methods 0.000 claims description 36
- 239000012528 membrane Substances 0.000 claims description 22
- 239000000843 powder Substances 0.000 claims description 22
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 20
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 19
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 19
- 241000628997 Flos Species 0.000 claims description 18
- CTSPAMFJBXKSOY-UHFFFAOYSA-N ellipticine Chemical compound N1=CC=C2C(C)=C(NC=3C4=CC=CC=3)C4=C(C)C2=C1 CTSPAMFJBXKSOY-UHFFFAOYSA-N 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 18
- 238000000108 ultra-filtration Methods 0.000 claims description 17
- 238000005341 cation exchange Methods 0.000 claims description 16
- 238000001914 filtration Methods 0.000 claims description 13
- 238000004140 cleaning Methods 0.000 claims description 9
- 238000001816 cooling Methods 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 238000000643 oven drying Methods 0.000 claims description 9
- 238000010298 pulverizing process Methods 0.000 claims description 9
- 239000003480 eluent Substances 0.000 claims description 8
- 239000002244 precipitate Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 5
- 239000011148 porous material Substances 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 238000007738 vacuum evaporation Methods 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 2
- 239000002246 antineoplastic agent Substances 0.000 claims description 2
- 229940041181 antineoplastic drug Drugs 0.000 claims description 2
- 239000003963 antioxidant agent Substances 0.000 claims description 2
- 230000003078 antioxidant effect Effects 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- ZIXGXMMUKPLXBB-UHFFFAOYSA-N Guatambuinine Natural products N1C2=CC=CC=C2C2=C1C(C)=C1C=CN=C(C)C1=C2 ZIXGXMMUKPLXBB-UHFFFAOYSA-N 0.000 claims 1
- SUYXJDLXGFPMCQ-INIZCTEOSA-N SJ000287331 Natural products CC1=c2cnccc2=C(C)C2=Nc3ccccc3[C@H]12 SUYXJDLXGFPMCQ-INIZCTEOSA-N 0.000 claims 1
- 238000007605 air drying Methods 0.000 claims 1
- 238000001291 vacuum drying Methods 0.000 claims 1
- 229920002678 cellulose Polymers 0.000 abstract description 15
- 239000001913 cellulose Substances 0.000 abstract description 15
- 125000002091 cationic group Chemical group 0.000 abstract description 13
- 102000004169 proteins and genes Human genes 0.000 abstract description 6
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 6
- 239000012535 impurity Substances 0.000 abstract description 5
- 239000004480 active ingredient Substances 0.000 abstract description 3
- 239000000341 volatile oil Substances 0.000 abstract description 3
- 229930013930 alkaloid Natural products 0.000 abstract description 2
- 229930003658 monoterpene Natural products 0.000 abstract description 2
- 150000002773 monoterpene derivatives Chemical class 0.000 abstract description 2
- 235000002577 monoterpenes Nutrition 0.000 abstract description 2
- 239000003960 organic solvent Substances 0.000 abstract description 2
- 150000008442 polyphenolic compounds Chemical class 0.000 abstract description 2
- 235000013824 polyphenols Nutrition 0.000 abstract description 2
- 238000005406 washing Methods 0.000 description 28
- 235000019441 ethanol Nutrition 0.000 description 22
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 21
- 229910052799 carbon Inorganic materials 0.000 description 15
- 230000000052 comparative effect Effects 0.000 description 15
- 238000012360 testing method Methods 0.000 description 15
- 230000007935 neutral effect Effects 0.000 description 14
- 238000002835 absorbance Methods 0.000 description 11
- 206010028980 Neoplasm Diseases 0.000 description 9
- 238000005119 centrifugation Methods 0.000 description 9
- 238000001035 drying Methods 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 210000000952 spleen Anatomy 0.000 description 8
- 241000699670 Mus sp. Species 0.000 description 7
- 239000008367 deionised water Substances 0.000 description 7
- 229910021641 deionized water Inorganic materials 0.000 description 7
- 239000012467 final product Substances 0.000 description 7
- 150000004676 glycans Chemical class 0.000 description 7
- 238000011068 loading method Methods 0.000 description 7
- 239000012982 microporous membrane Substances 0.000 description 6
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 6
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 4
- 238000005374 membrane filtration Methods 0.000 description 4
- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 description 4
- 150000003254 radicals Chemical class 0.000 description 4
- 230000002000 scavenging effect Effects 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 241001303601 Rosacea Species 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 230000007760 free radical scavenging Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 201000004700 rosacea Diseases 0.000 description 3
- 229960004889 salicylic acid Drugs 0.000 description 3
- 238000010998 test method Methods 0.000 description 3
- WRMNZCZEMHIOCP-UHFFFAOYSA-N 2-phenylethanol Chemical compound OCCC1=CC=CC=C1 WRMNZCZEMHIOCP-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- GLZPCOQZEFWAFX-UHFFFAOYSA-N Geraniol Chemical compound CC(C)=CCCC(C)=CCO GLZPCOQZEFWAFX-UHFFFAOYSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 2
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 description 2
- 235000004789 Rosa xanthina Nutrition 0.000 description 2
- 206010039491 Sarcoma Diseases 0.000 description 2
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000004305 biphenyl Substances 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 229920003086 cellulose ether Polymers 0.000 description 2
- QMVPMAAFGQKVCJ-UHFFFAOYSA-N citronellol Chemical compound OCCC(C)CCC=C(C)C QMVPMAAFGQKVCJ-UHFFFAOYSA-N 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 2
- RRAFCDWBNXTKKO-UHFFFAOYSA-N eugenol Chemical compound COC1=CC(CC=C)=CC=C1O RRAFCDWBNXTKKO-UHFFFAOYSA-N 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 229940075930 picrate Drugs 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 229940079877 pyrogallol Drugs 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000005728 strengthening Methods 0.000 description 2
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 2
- QMVPMAAFGQKVCJ-SNVBAGLBSA-N (R)-(+)-citronellol Natural products OCC[C@H](C)CCC=C(C)C QMVPMAAFGQKVCJ-SNVBAGLBSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- NPBVQXIMTZKSBA-UHFFFAOYSA-N Chavibetol Natural products COC1=CC=C(CC=C)C=C1O NPBVQXIMTZKSBA-UHFFFAOYSA-N 0.000 description 1
- 241001391944 Commicarpus scandens Species 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 239000005770 Eugenol Substances 0.000 description 1
- 206010019133 Hangover Diseases 0.000 description 1
- GLZPCOQZEFWAFX-JXMROGBWSA-N Nerol Natural products CC(C)=CCC\C(C)=C\CO GLZPCOQZEFWAFX-JXMROGBWSA-N 0.000 description 1
- 231100000674 Phytotoxicity Toxicity 0.000 description 1
- UVMRYBDEERADNV-UHFFFAOYSA-N Pseudoeugenol Natural products COC1=CC(C(C)=C)=CC=C1O UVMRYBDEERADNV-UHFFFAOYSA-N 0.000 description 1
- 235000011449 Rosa Nutrition 0.000 description 1
- 241000109329 Rosa xanthina Species 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- 244000178231 Rosmarinus officinalis Species 0.000 description 1
- 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 1
- 239000013543 active substance Substances 0.000 description 1
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 229930002877 anthocyanin Natural products 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 235000013734 beta-carotene Nutrition 0.000 description 1
- 239000011648 beta-carotene Substances 0.000 description 1
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 1
- JGQFVRIQXUFPAH-UHFFFAOYSA-N beta-citronellol Natural products OCCC(C)CCCC(C)=C JGQFVRIQXUFPAH-UHFFFAOYSA-N 0.000 description 1
- 229960002747 betacarotene Drugs 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000000484 citronellol Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- CGEBIRQTPVFDNU-UHFFFAOYSA-N diazanium ethanol sulfate hydrate Chemical compound [NH4+].[NH4+].O.CCO.[O-]S([O-])(=O)=O CGEBIRQTPVFDNU-UHFFFAOYSA-N 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229960002217 eugenol Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000010685 fatty oil Substances 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229910000358 iron sulfate Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000002175 menstrual effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000001054 red pigment Substances 0.000 description 1
- BTIHMVBBUGXLCJ-OAHLLOKOSA-N seliciclib Chemical compound C=12N=CN(C(C)C)C2=NC(N[C@@H](CO)CC)=NC=1NCC1=CC=CC=C1 BTIHMVBBUGXLCJ-OAHLLOKOSA-N 0.000 description 1
- 206010041569 spinal fracture Diseases 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Toxicology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The application discloses a method for obtaining rose glycoside by low-frequency ultrasound, which comprises the following steps: (1) raw material pretreatment; (2) ultrasonic extraction of raw materials; (3) treating the mixed solution; and (4) treating the extracting solution and (5) decoloring and refining. According to the application, the rose is dried and crushed, so that the contact area between the rose and the extractant is enlarged, and the extraction rate of the rose glycoside is improved. Meanwhile, polyethylene glycol-ammonium sulfate-water is used as an extracting agent, so that the volatile oil, monoterpenes, polyphenols, alkaloids and other substances of the active ingredients in the rose can be carried out, the polyethylene glycol can be evaporated and removed under reduced pressure, and no organic solvent residue is left, so that the safety is ensured. Dissolving the extract in a mixed solution of ethanol and sodium chloride, and quickly passing through cationic cellulose at room temperature to adsorb impurity proteins on the cationic cellulose and then removing the impurity proteins. The application has the advantages of rapidness, high efficiency, simple operation and the like, and has higher application value and development prospect.
Description
Technical Field
The application relates to the field of plant glycoside extraction, in particular to a method for obtaining rose glycoside by low-frequency ultrasound.
Background
Roses are the generic name for a variety of plants and cultivated flowers of the rosaceae, rosa. The modern medical community considers that the rose has the effects of strengthening liver, relieving stuffiness, resolving depression, detoxifying, strengthening lung, removing phytotoxicity caused by antibiotics, balancing endocrine, relieving nerve fatigue, hangover and the like. The rose contains a plurality of volatile components such as volatile oil, citronellol, nerol, eugenol, phenethyl alcohol and other substances with fragrance, also contains canopy, boots, wax, fatty oil, organic acid, rose rate red pigment, beta-carotene, amino acid, a plurality of vitamins and trace elements, plays a great role in regulating qi and resolving depression, regulating blood and dispelling blood stasis, astringing and relieving diarrhea and softening liver and restoring consciousness, and anthocyanin, flavone, polysaccharide and nucleic acid in the rose can be recycled. Glycosides, also known as glycoside compounds, are compounds formed by linking a terminal carbon atom of a sugar or sugar derivative to another class of non-sugar substances, known as aglycones, ligands or aglycones. The rosacea is also called as the rosacea, and the rosacea is recognized by the food and medical community as a physiologically active substance and a specific health care effect. The effect of the rose glycoside is relatively wide, and the rose glycoside not only can moisten skin and nourish face, reduce fat and lose weight, but also can relieve constipation by proper drinking of the rose glycoside with the functions of promoting blood circulation to remove blood stasis, has a large amount of cellulose, can promote gastrointestinal peristalsis, and achieves the effects of reducing fat and losing weight. The rose glycoside also has the effects of promoting blood circulation and replenishing blood, and can be used for regulating female irregular menstruationAnd menstrual abdominal pain. The existing extraction method of the rose glycoside has the following defects: (1) water extraction and alcohol precipitation method: the time consumption is long, the extraction rate is low, and the long-time extraction can also have a certain influence on the activity and structure of polysaccharide; (2) microwave method: the polysaccharide structure is easy to break; (3) enzymatic extraction: the cost of enzyme extraction by an enzyme method is high, and the cost of enzyme extraction by an industrial method is too high to be suitable for mass production; (4) Supercritical CO 2 Extraction: has high requirements on operation technology. Therefore, a method for obtaining the rose glycoside by low-frequency ultrasonic waves, which is simple to operate and high in extraction rate, is urgently needed.
Disclosure of Invention
Aiming at the defects in the prior art, the application provides a method for obtaining the ellipticin by low-frequency ultrasonic waves, which aims to solve the problems in the prior art.
In order to achieve the above purpose, the present application provides the following technical solutions: a method for obtaining a rose glycoside by low-frequency ultrasonic, which comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1: 5-8, uniformly stirring, and performing ultrasonic extraction for 20-30 min to obtain a mixed solution; wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:3-5: 7-9, wherein the ultrasonic is low-frequency ultrasonic, the ultrasonic power is 1-3 kW, the ultrasonic frequency is 20-28 kHz, and the extraction temperature is 35-45 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 1-3 times, and completely combining the supernatant; sequentially filtering the combined supernatant by vacuum evaporation and ultrafiltration membrane, and recovering polyethylene glycol and ammonium sulfate to obtain an extract; wherein the centrifugal speed is 2000-4000 rpm, the centrifugal time is 10-15 min, the reduced pressure evaporation pressure is 0.03-0.05 MPa, the reduced pressure evaporation temperature is 200-260 ℃, and the reduced pressure evaporation time is 8-10 min;
s4, treating the extracting solution: adding an ethanol aqueous solution and a sodium chloride aqueous solution which are 4-6 times of the total mass into the extract obtained in the step S3, wherein the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 2-4:1, heating to 45-50 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with an absolute ethanol solution through a cation exchange chromatographic column, collecting and combining eluents, and concentrating under reduced pressure to obtain a target extract;
s5, decoloring and refining: adding powdered activated carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the activated carbon is 0.1-0.3 g/mL, and uniformly mixing, and then carrying out constant-temperature water bath for 1-3 h; centrifuging, taking supernatant, carrying out suction filtration for 1-3 times by using a microporous filter membrane, and removing active carbon residues to obtain the rose glycoside; wherein the centrifugal speed is 1000-3000 rpm, and the pore size of the microporous filter membrane is 0.22-0.70 mu m.
Further, the rose used in the present application is a Bulgarian variety from Hainan Rose valley industries development Co.
Further, the polyethylene glycol was PEG4000, commercially available from Shanghai chain collection chemical Co., ltd.
Further, in the step S2, the ultrasonic equipment is an ultrasonic extraction machine Y-TC-5, which is purchased from Shanghai Yu inkstone mechanical equipment Co.
Further, mixing rose powder and an extracting agent in the step S2 according to a mass ratio of 1:7, and performing ultrasonic extraction for 25min, wherein the mass ratio of polyethylene glycol, ammonium sulfate and water is 1:4:8, the ultrasonic power is 2kW, the ultrasonic frequency is 25kHz, and the extraction temperature is 40 ℃.
Further, the model of the centrifuge in the step S3 is TD-5M, which is purchased from the biotechnology company of Jinan Tian Ling; the reduced pressure evaporator was a rotary evaporator RE-10L available from Zhengzhou scale equipment Co., ltd.
Further, in the step S3, the ultrasonic extraction is carried out for 2 times, the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, the reduced pressure evaporation time is 9min, and the ultrafiltration membrane size is 0.01 μm.
Further, in the step S4, the mass concentration of the ethanol aqueous solution and the sodium chloride aqueous solution is 5 times of the total mass, the mass concentration of the ethanol aqueous solution is 95%, the mass concentration of the sodium chloride aqueous solution is 0.1mol/L, the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 3:1, and the mixture is heated at 48 ℃ and stirred uniformly. The cation exchange chromatographic column is prepared by dissolving 8g of cationic cellulose in 50mL of deionized water, and loading into column. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Further, the cationic cellulose is a chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether available from Jining Tang Yi chemical Co.
Further, the addition amount of the activated carbon in the step S5 is 0.2g/mL, and after the mixture is uniformly mixed, the constant temperature is 45 ℃, and the water bath is performed for 2 hours; and taking supernatant after centrifugation, and carrying out suction filtration for 2 times by using a microporous filter membrane to remove active carbon residues, wherein the pore size of the microporous filter membrane is 0.45 mu m, the centrifugation rotating speed is 2000rpm, and the microporous filter membrane is purchased from Guangdong CycloKai microorganism technology Co.
Still further, the rose is any one of fresh rose or rose dreg.
Furthermore, the extracted rose glycoside is applied to the preparation of antioxidant drugs.
Furthermore, the application of the extracted rose glycoside in preparing antitumor drugs
Compared with the prior art, the application has the beneficial effects that: the application provides a method for obtaining the rose glycoside by low-frequency ultrasonic, which is used for drying and crushing rose flowers, expanding the contact area with an extracting agent and being beneficial to improving the extraction rate of the rose glycoside. The polyethylene glycol-ammonium sulfate is used as an extracting agent, so that the volatile oil, monoterpenes, polyphenols, alkaloids and other substances of the active ingredients in the rose can be carried out, the polyethylene glycol can be evaporated and removed, and no organic solvent residue is left to ensure safety. The concentrated solution is dissolved in the mixed solution of ethanol and sodium chloride, and under the condition of room temperature, the concentrated solution rapidly passes through the cationic cellulose, so that proteins are adsorbed on the cationic cellulose and then removed, impurity proteins are effectively removed, and the purity of the rose glycoside is ensured. The application can fully extract the effective components in the rose dreg and avoid waste.
Detailed Description
The technical scheme of the application is further described in detail below with reference to the specific embodiments.
The rose used in this experiment was a Bulgarian variety from Hainan Rose grain industry development Co.
The polyethylene glycol was PEG4000, commercially available from Shanghai chain collection chemical Co.
The ultrasonic equipment is an ultrasonic extraction machine Y-TC-5, which is purchased from Shanghai Yu inkstone mechanical equipment Co.
The model of the centrifuge is TD-5M, and is purchased from Biotechnology limited company of Jinan Tiankou; the reduced pressure evaporator was a rotary evaporator RE-10L available from Zhengzhou scale equipment Co., ltd.
Microporous filters were purchased from Guangdong CycloKai microbiological technologies Co.
The cationic cellulose is chlorinated-2-hydroxy-3- (trimethylamino) propyl polyethylene oxide cellulose ether, available from Jining Tang Yi chemical Co.
Example 1
The method for obtaining the rose glycoside by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1:5, mixing, stirring uniformly, and performing ultrasonic extraction for 20min to obtain a mixed solution; wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:3:7, ultrasonic is low-frequency ultrasonic, ultrasonic power is 1kW, ultrasonic frequency is 20kHz, and extraction temperature is 35 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out the supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 1 time, and completely combining the supernatants; and (3) sequentially carrying out reduced pressure evaporation and ultrafiltration membrane filtration on the combined supernatant, and recovering polyethylene glycol and ammonium sulfate to obtain an extracting solution. Wherein the centrifugal speed is 2000rpm, the centrifugal time is 10min, the reduced pressure evaporation pressure is 0.03MPa, the reduced pressure evaporation temperature is 200 ℃, the reduced pressure evaporation time is 8min, and the ultrafiltration membrane size is 0.01 μm.
S4, treating the extracting solution: adding 95% ethanol aqueous solution and 0.1mol/L sodium chloride aqueous solution with the mass concentration of 4 times of the total mass into the extracting solution obtained in the step S3, heating to 45 ℃ and uniformly stirring and mixing, cooling to room temperature, eluting with absolute ethanol solution by using a cation exchange chromatographic column, collecting and combining the eluents, and concentrating under reduced pressure to obtain a target extracting solution;
s5, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the active carbon is 0.1g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 1h; centrifuging, collecting supernatant, and vacuum filtering with 0.22 μm microporous membrane for 1 time to remove active carbon residue to obtain the final product. Wherein the centrifugation speed is 1000rpm.
The cation exchange chromatographic column is prepared by dissolving 5g of cationic cellulose in 30mL of deionized water, and loading. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Example 2
The method for obtaining the rose glycoside by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1:8, mixing, stirring uniformly, and performing ultrasonic extraction for 30min to obtain a mixed solution. Wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:5:9, ultrasonic is low-frequency ultrasonic, ultrasonic power is 3kW, ultrasonic frequency is 28kHz, and extraction temperature is 45 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out the supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 3 times, and combining all the supernatants. Sequentially filtering the combined supernatant by vacuum evaporation and ultrafiltration membrane, and recovering polyethylene glycol and ammonium sulfate to obtain an extract; wherein the centrifugal speed is 4000rpm, the centrifugal time is 15min, the reduced pressure evaporation pressure is 0.05MPa, the reduced pressure evaporation temperature is 260 ℃, the reduced pressure evaporation time is 10min, and the ultrafiltration membrane size is 0.01 mu m;
s4, treating the extracting solution: adding 95% ethanol aqueous solution and 0.1mol/L sodium chloride aqueous solution with the mass concentration of 6 times of the total mass into the extracting solution obtained in the step S3, heating to 50 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with absolute ethanol solution by using a cation exchange chromatographic column, collecting and combining the eluents, and concentrating under reduced pressure to obtain a target extracting solution;
s5, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the active carbon is 0.3g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 3 hours; centrifuging, collecting supernatant, and vacuum filtering with 0.70 μm microporous membrane for 3 times to remove active carbon residue to obtain the final product. Wherein the centrifugation speed was 3000rpm.
The cation exchange chromatographic column is prepared by dissolving 10g of cationic cellulose in 60mL of deionized water, and loading into column. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Example 3
The method for obtaining the rose glycoside by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1:7, mixing, stirring uniformly, and performing ultrasonic extraction for 25min to obtain a mixed solution. Wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:4:8, ultrasonic is low-frequency ultrasonic, ultrasonic power is 2kW, ultrasonic frequency is 25kHz, and extraction temperature is 40 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out the supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 2 times, and combining all the supernatants. And (3) sequentially carrying out reduced pressure evaporation and ultrafiltration membrane filtration on the combined supernatant, and recovering polyethylene glycol and ammonium sulfate to obtain an extracting solution. Wherein the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, the reduced pressure evaporation time is 9min, and the ultrafiltration membrane size is 0.01 μm.
S4, treating the extracting solution: adding 95% ethanol aqueous solution and 0.1mol/L sodium chloride aqueous solution with the mass concentration of 5 times of the total mass into the extracting solution obtained in the step S3, heating to 48 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with absolute ethanol solution by using a cation exchange chromatographic column, collecting and combining the eluents, and concentrating under reduced pressure to obtain a target extracting solution;
s5, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the active carbon is 0.2g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 2 hours; centrifuging, collecting supernatant, and vacuum filtering with 0.45 μm microporous membrane for 2 times to remove active carbon residue to obtain the final product. Wherein the centrifugation speed was 2000rpm.
The cation exchange chromatographic column is prepared by taking 8g of cationic cellulose, adding 50mL of deionized water, dissolving and loading into a column. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Comparative example 1
The difference between this comparative example and example 3 is that: the ultrasonic extraction step is not needed, and the contents of other steps and components are unchanged.
The method for obtaining the ellipticin by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1:7, mixing, stirring uniformly, centrifuging, filtering the combined supernatant by a decompression evaporation and ultrafiltration membrane in sequence, and recovering polyethylene glycol and ammonium sulfate to obtain an extract; wherein the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, the reduced pressure evaporation time is 9min, and the ultrafiltration membrane size is 0.01 μm;
s3, treating an extracting solution: adding 95% ethanol aqueous solution with a mass concentration of 5 times of the total mass and 0.1mol/L sodium chloride aqueous solution into the extracting solution obtained in the step S2, wherein the volume ratio of the ethanol aqueous solution to the sodium chloride is 3:1, heating to 48 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with absolute ethanol solution through a cation exchange chromatographic column, collecting and combining eluents, and concentrating under reduced pressure to obtain a target extract;
s4, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S3, wherein the adding amount of the active carbon is 0.2g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 2 hours; centrifuging, collecting supernatant, and vacuum filtering with 0.45 μm microporous membrane for 2 times to remove active carbon residue to obtain the final product. Wherein the centrifugation speed was 2000rpm.
The cation exchange chromatographic column is prepared by taking 8g of cationic cellulose, adding 50mL of deionized water, dissolving and loading into a column. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Comparative example 2
The difference between this comparative example and example 3 is that: the concentrated solution has no cation exchange chromatographic column eluting step, and the rest steps and component content are unchanged.
The method for obtaining the ellipticin by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1:7, mixing, stirring uniformly, and performing ultrasonic extraction for 25min to obtain a mixed solution. Wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:4:8, ultrasonic is low-frequency ultrasonic, ultrasonic power is 2kW, ultrasonic frequency is 25kHz, and extraction temperature is 40 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out the supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 2 times, and completely combining the supernatants; . And (3) sequentially carrying out reduced pressure evaporation and ultrafiltration membrane filtration on the combined supernatant, and recovering polyethylene glycol and ammonium sulfate to obtain an extracting solution. The method comprises the steps of carrying out a first treatment on the surface of the Wherein the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, the reduced pressure evaporation time is 9min, and the ultrafiltration membrane size is 0.01 μm;
s4, treating the extracting solution: adding 95% ethanol aqueous solution with a mass concentration of 5 times of the total mass and 0.1mol/L sodium chloride aqueous solution into the extracting solution obtained in the step S3, wherein the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 3:1, heating to 48 ℃, stirring and mixing uniformly, cooling to room temperature, and concentrating under reduced pressure to obtain a target extract;
s5, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the active carbon is 0.2g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 2 hours; centrifuging, collecting supernatant, and vacuum filtering with 0.45 μm microporous membrane for 2 times to remove active carbon residue to obtain the final product. Wherein the centrifugation speed was 2000rpm.
Comparative example 3
The difference between this comparative example and example 3 is that: the decoloring and refining process has no microporous filtering step, and the other steps and the component content are unchanged.
The method for obtaining the ellipticin by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1:7, mixing, stirring uniformly, and performing ultrasonic extraction for 25min to obtain a mixed solution. Wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:4:8, ultrasonic is low-frequency ultrasonic, ultrasonic power is 2kW, ultrasonic frequency is 25kHz, and extraction temperature is 40 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out the supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 2 times, and combining all the supernatants. Sequentially filtering the combined supernatant by vacuum evaporation and ultrafiltration membrane, and recovering polyethylene glycol and ammonium sulfate to obtain an extract; wherein the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, the reduced pressure evaporation time is 9min, and the ultrafiltration membrane size is 0.01 μm;
s4, treating the extracting solution: adding 95% ethanol aqueous solution with a mass concentration of 5 times of the total mass and 0.1mol/L sodium chloride aqueous solution into the extracting solution obtained in the step S3, wherein the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 3:1, heating to 48 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with absolute ethanol solution through a cation exchange chromatographic column, collecting and combining eluents, and concentrating under reduced pressure to obtain a target extract;
s5, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the active carbon is 0.2g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 2 hours; centrifuging, and collecting supernatant to obtain the final product. Wherein the centrifugation speed was 2000rpm.
The cation exchange chromatographic column is prepared by taking 8g of cationic cellulose, adding 50mL of deionized water, dissolving and loading into a column. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Comparative example 4
The difference between this comparative example and example 3 is that: in the step S2, ethanol is used as the ultrasonic extractant instead of polyethylene glycol, and the content of the rest steps and components is unchanged.
The method for obtaining the ellipticin by low-frequency ultrasonic comprises the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally drying, oven drying in vacuum oven at 80deg.C, and pulverizing into 70 mesh flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: ethanol-ammonium sulfate-water is used as an extracting agent, and the rose powder obtained in the step S1 and the extracting agent are mixed according to the mass ratio of 1:7, mixing, stirring uniformly, and performing ultrasonic extraction for 25min to obtain a mixed solution. Wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:4:8, ultrasonic is low-frequency ultrasonic, ultrasonic power is 2kW, ultrasonic frequency is 25kHz, and extraction temperature is 40 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out the supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 2 times, and combining all the supernatants. And (3) sequentially carrying out reduced pressure evaporation and ultrafiltration membrane filtration on the combined supernatant, and recovering polyethylene glycol and ammonium sulfate to obtain an extracting solution. Wherein the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, and the reduced pressure evaporation time is 9min; the ultrafiltration membrane size was 0.01. Mu.m.
S4, treating the extracting solution: adding 95% ethanol aqueous solution with a mass concentration of 5 times of the total mass and 0.1mol/L sodium chloride aqueous solution into the extracting solution obtained in the step S3, wherein the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 3:1, heating to 48 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with absolute ethanol solution through a cation exchange chromatographic column, collecting and combining eluents, and concentrating under reduced pressure to obtain a target extract;
s5, decoloring and refining: adding powdery active carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the active carbon is 0.2g/mL, and uniformly mixing, and then carrying out constant-temperature water bath at 45 ℃ for 2 hours; centrifuging, collecting supernatant, and vacuum filtering with 0.45 μm microporous membrane for 2 times to remove active carbon residue to obtain the final product. Wherein the centrifugation speed was 2000rpm.
The cation exchange chromatographic column is prepared by taking 8g of cationic cellulose, adding 50mL of deionized water, dissolving and loading into a column. Firstly, adding 0.2mmol/L NaOH solution for washing, adding water for washing to be neutral, then adding 0.2mmol/L HCl for washing, adding water for washing to be neutral, and preserving for later use.
Test example 1 extraction yield of ellipticin
1.1 test materials
The rose glycoside prepared in examples 1 to 3 and comparative examples 1 to 4 was used.
Alpha-1502 ultraviolet-visible spectrophotometer: purchased from Shanghai Spectrum instruments Co., ltd
1.2 test methods
Accurately sucking 4.0mL of the rose glycoside extract in a 100mL volumetric flask, metering the volume to a scale with distilled water, and mixing uniformly. Absorbance was measured at 490 nm. The calculation formula is as follows:
wherein: c is mass concentration of the rose glycoside, mg/mL; n is dilution multiple; v is the volume mL of the constant volume of the rose glycoside extracting solution; m is the mass of rose powder, g.
1.3 test results
TABLE 1
| Sequence number | Example 1 | Example 2 | Example 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Extraction rate | 70.45％ | 68.12％ | 78.56％ | 20.26％ | 59.75％ | 61.16％ | 65.12％ |
1.4 test results
As can be seen from table 1, in the method for obtaining the rose glycoside by low-frequency ultrasound, the extraction rate of the rose glycoside in example 3 is highest, which reaches 78.56%; as can be obtained by comparing example 3 with comparative examples 1 to 4, respectively, the rose powder was not subjected to ultrasonic treatment, and the active ingredient in the rose could not be extracted, resulting in a decrease in the extraction rate of the rose glycoside; the concentrated solution is not eluted by an ion cellulose column, so that the concentrated solution contains a large amount of impurity proteins, and the extraction rate of the rose glycoside is affected; the target extracting solution is not subjected to microporous filtration and contains a large amount of pigments, so that the extraction rate of the rose glycoside is reduced; the extraction rate of the polyethylene glycol used as the ultrasonic extractant is better than that of ethanol.
Test example 2 in vitro oxidation of rosmarin
2.1 test materials and reagents
Using the rose glycoside prepared in example 3 and comparative examples 1 to 4, a commercial rose glycoside (available from vickers biotechnology limited, sichuan province, CAS No. 54835-70-0) was used as a positive control group; absolute ethyl alcohol: purchased from Cangzhou Heyue chemical Co., ltd; 1, 2-diphenyl picrate hydrazyl: purchased from Sigma, usa; iron sulfate: purchased from Tianjin chemical reagent-works; salicylic acid: purchased from Shanghai pacifying business Co., ltd; h 2 O 2 Aqueous solution: purchased from Tianjin, inc. of the chemical industry;
2.2 test instruments
Alpha-1502 ultraviolet-visible spectrophotometer: purchased from Shanghai Spectrum instruments Co., ltd
2.3 test methods
(1) Determination of the ability to scavenge 1, 2-diphenyl picrate hydrazyl radical (DPPH-): preparing 2mg/mL of rose glycoside solution and 0.2mmol/L of DPPH solution with absolute ethyl alcohol, respectively taking 2mL of rose glycoside solution and DPPH solution, placing in a test tube, shaking uniformly, standing at room temperature for 30min, and obtaining absorbance A 1 The method comprises the steps of carrying out a first treatment on the surface of the Simultaneously measuring absorbance A after mixing 2mL DPPH solution and 2mL distilled water 2 And absorbance of 2mL of rose glycoside solution mixed with 2mL of absolute ethyl alcohol is A 3 And (5) carrying out light-shielding reaction. At 517nm, the absorbance was measured, and the clearance was calculated as:
(2) Determination of the ability to scavenge hydroxyl (-OH) radicals: the test adopts a salicylic acid method, and 9mmol/L FeSO is added into a colorimetric tube in sequence 4 2mL,9mmol/L salicylic acid 2mL, ethanol 2mL, 2mL of a rose glycoside solution, 8.8mmol/L H 2 O 2 2mL, where H 2 O 2 The whole reaction was started, and after 0.5h at 37℃the absorbance A of the sample was measured at 510nm 1 Measuring absorbance A by taking distilled water instead of the rose glycoside solution as blank for zeroing 0 The formula is as follows:
(3) Determination of the ability to scavenge superoxide anion radical: adopting a pyrogallol oxidation method, taking 4.5mL of 0.05mol/L, pH Tris-HCL buffer solution in a test tube, and measuring absorbance A in a water bath at 25 ℃ for 20min 0 Adding a solution of rose glycoside, and measuring absorbance A 1 Then 0.5mL of a solution of pyrogallol with the concentration of 2.5mmol/L is added, the mixture is shaken and uniformly mixed in a water bath with the temperature of 25 ℃ for 5min, and the reaction is immediately stopped by 0.1mL of a solution of HCL with the concentration of 8mmol/L, and the absorbance A is measured 2 The absorbance was measured at 320nm, and the formula was:
2.4 test results
TABLE 2
As is clear from Table 2, the extraction rate of the rose glycoside is positively correlated with the scavenging ability of the free radical, and example 3 has good scavenging ability to DPPH free radical scavenging rate, hydroxyl free radical scavenging rate and superoxide anion free radical scavenging rate, and the scavenging ability to the free radical is reduced due to the influence of the existence of impurity proteins and pigments, and the scavenging ability of the rose glycoside prepared in example 3 is slightly better than that of the commercially available rose glycoside by using polyethylene glycol as an ultrasonic extractant rather than ethanol.
Test example 3 anti-tumor and spleen index Effect of roscovitine
3.1 test materials
The rose glycoside prepared in example 3 was used; 50 swiss mice, 10 per group.
S180 mouse sarcoma strain: purchased from Shanghai Qincheng Biotech Inc.
3.2 test methods
Taking S180 mouse sarcoma strain, diluting with physiological saline to 5×10 6 The single cell suspension/mL was centrifuged at 3500r/min for 10min, the supernatant was discarded, and diluted 1X 10 with physiological saline 10 The mice were inoculated with 0.2mL of a suspension on the sterilized right side underarm and on the subcutaneous side of the arm to form solid tumors. Wherein the blank group is not connected with tumor cells; the negative control group only receives tumor cells, and does not inject the rose glycoside; the test groups were inoculated with tumor cells and injected with the amounts of rose glycoside according to Table 3.
TABLE 3 Table 3
Note that: blank control group, no tumor; other groups all receive S180 tumor cells
After 10 days of rose glycoside injection, each group of mice was weighed, sacrificed by vertebral fracture, spleens were removed, and solid tumors were peeled off.
(1) The anti-tumor inhibition rate calculation method comprises the following steps:
wherein C-average tumor weight (g) of control group
Average tumor weight (g) of T-injection group
(2) Calculation formula of spleen index:
spleen index = spleen weight (mg)/body weight (g)
3.3 test results
TABLE 4 Rose glycoside pair S 180 Influence of the tumor inhibition in mice
As can be seen from Table 4, with increasing dose of the injected rose glycoside, the tumor inhibition effect is more and more obvious, and the tumor of the mice in the negative control group is obviously enlarged. The three different dose groups (low, medium and high) of the rose glycoside have different degrees of inhibition on the armpit tumors, the inhibition rate of the low dose group is 35.62%, the inhibition rate of the medium dose group is 40.15%, and the inhibition rate of the high dose group is 72.48%.
TABLE 5 Rose glycoside pair S 180 Influence of the inhibition of the spleen index in mice
As can be seen from table 5, the spleen index of the mice in the three different dose groups was higher than that in the negative control group and the blank group, and the spleen index was increased with the increase of the dose, showing a significant dose-effect relationship.
The foregoing description of the preferred embodiments of the application is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the application.
Claims (9)
1. The method for obtaining the ellipticin by low-frequency ultrasonic is characterized by comprising the following steps:
s1, pretreatment of raw materials: cleaning flos Rosae Rugosae, naturally air drying, oven drying in vacuum drying oven, and pulverizing into flos Rosae Rugosae powder;
s2, ultrasonic extraction of raw materials: taking polyethylene glycol-ammonium sulfate-water as an extracting agent, and mixing the rose powder obtained in the step S1 with the extracting agent according to the mass ratio of 1: 5-8, uniformly stirring, and performing ultrasonic extraction for 20-30 min to obtain a mixed solution; wherein the mass ratio of polyethylene glycol to ammonium sulfate to water is 1:3 to 5: 7-9, wherein the ultrasonic power is 1-3 kW, the ultrasonic frequency is 20-28 kHz, and the extraction temperature is 35-45 ℃;
s3, treating the mixed solution: centrifuging the mixed solution obtained in the step S2, taking out supernatant, adding polyethylene glycol into the precipitate, performing ultrasonic extraction for 1-3 times, and completely combining the supernatant; sequentially filtering the combined supernatant by vacuum evaporation and ultrafiltration membrane, and recovering polyethylene glycol and ammonium sulfate to obtain an extract; wherein the centrifugal speed is 2000-4000 rpm, the centrifugal time is 10-15 min, the reduced pressure evaporation pressure is 0.03-0.05 MPa, the reduced pressure evaporation temperature is 200-260 ℃, and the reduced pressure evaporation time is 8-10 min;
s4, treating the extracting solution: adding an ethanol aqueous solution and a sodium chloride aqueous solution which are 4-6 times of the total mass into the extract obtained in the step S3, wherein the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 2-4:1, heating to 45-50 ℃, stirring and mixing uniformly, cooling to room temperature, eluting with an absolute ethanol solution through a cation exchange chromatographic column, collecting and combining eluents, and concentrating under reduced pressure to obtain a target extract;
s5, decoloring and refining: adding powdered activated carbon into the target extracting solution obtained in the step S4, wherein the adding amount of the activated carbon is 0.1-0.3 g/mL, and uniformly mixing, and then carrying out constant-temperature water bath for 1-3 h; centrifuging, taking supernatant, carrying out suction filtration for 1-3 times by using a microporous filter membrane, and removing active carbon residues to obtain the rose glycoside; wherein the centrifugal speed is 1000-3000 rpm, and the pore size of the microporous filter membrane is 0.22-0.70 mu m.
2. The method for obtaining the rose glycoside by low-frequency ultrasound according to claim 1, wherein in the step S2, rose powder and an extracting agent are mixed according to a mass ratio of 1:7, the ultrasonic extraction is carried out for 25min, and the mass ratio of polyethylene glycol, ammonium sulfate and water is 1:4:8, the ultrasonic power is 2kW, the ultrasonic frequency is 25kHz, and the extraction temperature is 40 ℃.
3. The method for obtaining the ellipticin by low-frequency ultrasonic according to claim 1, wherein in the step S3, the ultrasonic extraction is carried out for 2 times, the centrifugal speed is 3000rpm, the centrifugal time is 12min, the reduced pressure evaporation pressure is 0.04MPa, the reduced pressure evaporation temperature is 250 ℃, and the reduced pressure evaporation time is 9min.
4. The method for obtaining the ellipticine by low-frequency ultrasound according to claim 1, wherein in the step S4, the mass concentration of the ethanol aqueous solution and the mass concentration of the sodium chloride aqueous solution are 5 times of the total mass, the mass concentration of the ethanol aqueous solution is 95%, the mass concentration of the sodium chloride aqueous solution is 0.1mol/L, the volume ratio of the ethanol aqueous solution to the sodium chloride aqueous solution is 3:1, and the mixture is heated at 48 ℃ and stirred and mixed uniformly.
5. The method for obtaining the rose glycoside by low-frequency ultrasound according to claim 1, wherein the adding amount of the activated carbon in the step S5 is 0.2g/mL, and after the mixture is uniformly mixed, the constant temperature is 45 ℃, and the constant temperature is carried out for 2 hours; and centrifuging, taking supernatant, and carrying out suction filtration for 2 times by using a microporous filter membrane to remove active carbon residues, wherein the centrifuging speed is 2000rpm, and the pore size of the microporous filter membrane is 0.45 mu m.
6. The method for obtaining a rose glycoside by low-frequency ultrasound according to claim 1, wherein the rose is either fresh rose or rose dreg.
7. A rose glycoside extracted by the method of any one of claims 1 to 6.
8. Use of the rose glycoside according to claim 7 in the preparation of an antioxidant medicament.
9. Use of the rose glycoside according to claim 7 in the preparation of an antitumor drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310833880.XA CN116925163A (en) | 2023-07-10 | 2023-07-10 | Method for obtaining rose glycoside by low-frequency ultrasound |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310833880.XA CN116925163A (en) | 2023-07-10 | 2023-07-10 | Method for obtaining rose glycoside by low-frequency ultrasound |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116925163A true CN116925163A (en) | 2023-10-24 |
Family
ID=88388813
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310833880.XA Pending CN116925163A (en) | 2023-07-10 | 2023-07-10 | Method for obtaining rose glycoside by low-frequency ultrasound |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116925163A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117586952A (en) * | 2024-01-19 | 2024-02-23 | 首都医科大学宣武医院 | Separation reagent and separation method for separating plasma exosomes |
-
2023
- 2023-07-10 CN CN202310833880.XA patent/CN116925163A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117586952A (en) * | 2024-01-19 | 2024-02-23 | 首都医科大学宣武医院 | Separation reagent and separation method for separating plasma exosomes |
| CN117586952B (en) * | 2024-01-19 | 2024-04-26 | 首都医科大学宣武医院 | Separation reagent and separation method for separating plasma exosomes |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109939027B (en) | Method for preparing ergothioneine-containing cosmetic stock solution by fermenting hericium erinaceus | |
| CN106117387B (en) | A kind of low molecular weight tremella polysaccharides and the preparation method and application thereof | |
| CN113150867B (en) | Preparation method of ganoderma lucidum extract oil rich in ganoderma lucidum triterpenes | |
| CN108823261B (en) | Ultralow-molecular-weight dendrobium officinale polysaccharide and preparation and application thereof | |
| CN112430546B (en) | Heterotrophic fermentation preparation method and application of chlamydomonas reinhardtii and chlamydomonas reinhardtii powder | |
| CN110128562B (en) | An antitumor fructus Psoraleae polysaccharide, its extraction and separation method, and its application in preparing antitumor drugs | |
| CN116925163A (en) | Method for obtaining rose glycoside by low-frequency ultrasound | |
| CN108250320B (en) | Low-ash ganoderma lucidum polysaccharide extract and preparation method thereof | |
| CN113024685A (en) | Low-molecular-weight Dictyophora indusiata (Vent. Ex pers) Fisch trum-Dictyophora (Vent. Ex pers) Fisch trum et Schott polysaccharide, and preparation method and application thereof | |
| CN111533680A (en) | Folium Mori extract and folium Mori multicomponent mixture with blood sugar lowering effect prepared from the same | |
| CN114105747A (en) | Method for improving curcumin extraction rate and purification effect | |
| CN110606900B (en) | Method for separating and purifying fructus Jujubae polysaccharide with antioxidant effect | |
| CN114146042A (en) | Saffron crocus fermentation product and preparation method and application thereof | |
| CN101229335B (en) | Enzyme method for preparing smilax scobinicaulis total saponin extract | |
| CN112480279A (en) | Low-molecular-weight sulfated polysaccharide of kelp and preparation and application thereof | |
| CN107119096B (en) | Preparation method and application of pholiota nameko active peptide | |
| CN107286264A (en) | The deep working method of Chinese date nutrient material separation | |
| CN109160954B (en) | Muskmelon eggplant acidic polysaccharide and purification method and application thereof | |
| CN110922499B (en) | Selenium-enriched sparassis crispa polysaccharide and preparation method and application thereof | |
| CN115594775B (en) | Preparation method and application of tremella polysaccharide | |
| CN114031695B (en) | Preparation process of high-purity shark chondroitin sulfate with anti-tumor effect | |
| CN113694152B (en) | Method for obtaining coix seed extracting solution by high-stability enzymolysis method | |
| CN112143769B (en) | A method for preparing radix Puerariae polypeptide extract from radix Puerariae residue and radix Puerariae polypeptide extract prepared thereby | |
| CN114632105A (en) | Comprehensive utilization method and application of peony seed meal | |
| KR20140102001A (en) | An method of Extracts for A GABA ingredient from germinated brown rice and that of using health function composition for a diabetic |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |