CN108191804A - The method of purification of Flavonoid substances Quercetin in a kind of Guava Leaf - Google Patents
The method of purification of Flavonoid substances Quercetin in a kind of Guava Leaf Download PDFInfo
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- CN108191804A CN108191804A CN201711466673.6A CN201711466673A CN108191804A CN 108191804 A CN108191804 A CN 108191804A CN 201711466673 A CN201711466673 A CN 201711466673A CN 108191804 A CN108191804 A CN 108191804A
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- guava leaf
- purification
- quercetin
- ultrafiltration
- solution
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- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 title claims abstract description 60
- 241000508269 Psidium Species 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 37
- 238000000746 purification Methods 0.000 title claims abstract description 35
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 title claims abstract description 30
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 title claims abstract description 30
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 title claims abstract description 30
- 229960001285 quercetin Drugs 0.000 title claims abstract description 30
- 235000005875 quercetin Nutrition 0.000 title claims abstract description 30
- 239000000126 substance Substances 0.000 title claims abstract description 20
- 229930003935 flavonoid Natural products 0.000 title claims abstract description 19
- 150000002215 flavonoids Chemical class 0.000 title claims abstract description 19
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 19
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 30
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 21
- 229930003944 flavone Natural products 0.000 claims abstract description 20
- 235000011949 flavones Nutrition 0.000 claims abstract description 20
- 239000000178 monomer Substances 0.000 claims abstract description 18
- 229920000344 molecularly imprinted polymer Polymers 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 150000002213 flavones Chemical class 0.000 claims abstract description 12
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 10
- 239000012043 crude product Substances 0.000 claims abstract description 10
- 238000001914 filtration Methods 0.000 claims abstract description 10
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 10
- 238000005119 centrifugation Methods 0.000 claims abstract description 9
- 230000009514 concussion Effects 0.000 claims abstract description 7
- 239000000284 extract Substances 0.000 claims abstract description 7
- 229920000642 polymer Polymers 0.000 claims abstract description 7
- 239000006228 supernatant Substances 0.000 claims abstract description 7
- 229920002554 vinyl polymer Polymers 0.000 claims abstract description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 26
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 239000003431 cross linking reagent Substances 0.000 claims description 12
- 239000003999 initiator Substances 0.000 claims description 10
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims description 9
- 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 claims description 8
- 239000000843 powder Substances 0.000 claims description 8
- OZAIFHULBGXAKX-UHFFFAOYSA-N 2-(2-cyanopropan-2-yldiazenyl)-2-methylpropanenitrile Chemical compound N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 claims description 7
- 238000001291 vacuum drying Methods 0.000 claims description 7
- ZCHPKWUIAASXPV-UHFFFAOYSA-N acetic acid;methanol Chemical compound OC.CC(O)=O ZCHPKWUIAASXPV-UHFFFAOYSA-N 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 239000000047 product Substances 0.000 claims description 6
- 238000012719 thermal polymerization Methods 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 150000002212 flavone derivatives Chemical class 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 230000008676 import Effects 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 2
- 238000000227 grinding Methods 0.000 claims description 2
- 238000006116 polymerization reaction Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- ICBJBNAUJWZPBY-UHFFFAOYSA-N 2-hydroxyethyl 3-methylbut-2-enoate Chemical group CC(=CC(=O)OCCO)C ICBJBNAUJWZPBY-UHFFFAOYSA-N 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 7
- 239000007788 liquid Substances 0.000 abstract description 5
- 238000007670 refining Methods 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract description 4
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 28
- 239000012528 membrane Substances 0.000 description 15
- 238000005516 engineering process Methods 0.000 description 12
- 238000000926 separation method Methods 0.000 description 11
- 229960000583 acetic acid Drugs 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- -1 Ketone compounds Chemical class 0.000 description 6
- 239000012460 protein solution Substances 0.000 description 6
- 239000012086 standard solution Substances 0.000 description 6
- DBCAQXHNJOFNGC-UHFFFAOYSA-N 4-bromo-1,1,1-trifluorobutane Chemical group FC(F)(F)CCCBr DBCAQXHNJOFNGC-UHFFFAOYSA-N 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 229940098773 bovine serum albumin Drugs 0.000 description 4
- 239000000287 crude extract Substances 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- STVZJERGLQHEKB-UHFFFAOYSA-N ethylene glycol dimethacrylate Substances CC(=C)C(=O)OCCOC(=O)C(C)=C STVZJERGLQHEKB-UHFFFAOYSA-N 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 239000012488 sample solution Substances 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000005977 Ethylene Substances 0.000 description 2
- 108010058846 Ovalbumin Proteins 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000000205 computational method Methods 0.000 description 2
- 238000010494 dissociation reaction Methods 0.000 description 2
- 230000005593 dissociations Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 229940092253 ovalbumin Drugs 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 239000001648 tannin Substances 0.000 description 2
- 235000018553 tannin Nutrition 0.000 description 2
- 229920001864 tannin Polymers 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 206010006458 Bronchitis chronic Diseases 0.000 description 1
- 206010007191 Capillary fragility Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- STECJAGHUSJQJN-GAUPFVANSA-N Hyoscine Natural products C1([C@H](CO)C(=O)OC2C[C@@H]3N([C@H](C2)[C@@H]2[C@H]3O2)C)=CC=CC=C1 STECJAGHUSJQJN-GAUPFVANSA-N 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 241000219926 Myrtaceae Species 0.000 description 1
- STECJAGHUSJQJN-UHFFFAOYSA-N N-Methyl-scopolamin Natural products C1C(C2C3O2)N(C)C3CC1OC(=O)C(CO)C1=CC=CC=C1 STECJAGHUSJQJN-UHFFFAOYSA-N 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 244000236580 Psidium pyriferum Species 0.000 description 1
- 235000013929 Psidium pyriferum Nutrition 0.000 description 1
- 244000294611 Punica granatum Species 0.000 description 1
- 235000014360 Punica granatum Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000000954 anitussive effect Effects 0.000 description 1
- 230000001088 anti-asthma Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
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- 230000009286 beneficial effect Effects 0.000 description 1
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- 206010006451 bronchitis Diseases 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000007451 chronic bronchitis Diseases 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 125000004356 hydroxy functional group Chemical group O* 0.000 description 1
- 230000001631 hypertensive effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- STECJAGHUSJQJN-FWXGHANASA-N scopolamine Chemical compound C1([C@@H](CO)C(=O)O[C@H]2C[C@@H]3N([C@H](C2)[C@@H]2[C@H]3O2)C)=CC=CC=C1 STECJAGHUSJQJN-FWXGHANASA-N 0.000 description 1
- 229960002646 scopolamine Drugs 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000003883 substance clean up Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a kind of methods of purification of Flavonoid substances Quercetin in Guava Leaf, include the following steps:(1) water is added in after Guava Leaf is crushed, is heated, filtering obtains guava leaf extract flavones crude product;(2) ethanol solution is dissolved in, supernatant is taken to carry out ultrafiltration cycle after filtering, obtains ultrafiltration membrance filter solution;(3) it is dissolved in acetonitrile using Quercetin monomer as template molecule, addition 4 vinyl of function monomer adjoins throat and prepares molecularly imprinted polymer;(4) ultrafiltration membrance filter solution is added in molecularly imprinted polymer, closed concussion is carried out in constant-temperature table, the Flavonoid substances Quercetin is obtained after centrifugation.The method of purification of the present invention, the feed liquid separated further refine purification with the imprinted polymer of synthesis again, obtain the Guava Leaf Flavonoid substances Quercetin of purifying, environmental protection simple for process, income at low cost is high, and refining effect is good.
Description
Technical field
The invention belongs to Flavonoid substances quercitrins in Guava Leaf compound purification field more particularly to a kind of Guava Leaf
The method of purification of element.
Background technology
Guava (Psidium guajava Linn.) Myrtaceae arbor originates in South America.In South China various regions plant
Training, common to have ease for wild species, north reaches the Anning Valley in Sichuan Province of South-west Sichuan, is born on wasteland or haugh;Fruit is edible;Ye Han
Volatile oil and tannin etc., hyoscine have stop dysentery, hemostasis, stomach invigorating and other effects;Ye Jing, which boils, removes tannin, dries and is used as tealeaves, taste
It is sweet, there is heat-clearing effect.The chemical constitution study of Guava Leaf is shown:Contain protein, polysaccharide, polyphenol, Huang in Guava Leaf
Ketone compounds and triterpene compound etc..
Flavone compound (flavonoids) is that one kind is present in nature, has 2- phenyl chromones
(flavone) compound of structure.There are one ketone carbonyl group in their molecules, the oxygen atom tool alkalinity on first, can with it is strong
Acid is into salt, and hydroxy derivatives have a yellow more, therefore also known as flavone or flavones.Flavone compound in plant usually with sugar
Glycoside is combined into, fraction exists in the form of free state (aglycon).All contain flavone compound in most plants,
It plant growth, develop, bloom, result and antibacterial diseases prevention etc. play an important role.
Quercetin also known as quercetin are one kind of flavone compound, and Quercetin is dissolved in glacial acetic acid, and alkaline aqueous solution is in
Yellow, is practically insoluble in water, and ethanol solution taste is very bitter.Drug can be used as, there is preferable eliminating the phlegm, antitussive action, and is had certain
Antiasthmatic effect.In addition with reduce blood pressure, enhance capillary resistance, reduce capillary fragility, reducing blood lipid, expansion hat
The effects that shape artery, increase coronary blood flow.For treating chronic bronchitis, also there is auxiliary to coronary heart disease and hypertensive patient
Therapeutic effect.
The separation of traditional guava leaf extract mainly uses solvent method, macroreticular resin absorbing method, microwave loss mechanisms, surpasses
Sound wave extraction method, supercritical extraction, but these method complex process, refining effect is poor, does not meet energy-saving and environment-friendly requirement, into
This height and income is low.And capillary electrophoresis, enzyme process are generally used for purifying micro or trace substance, to purification condition requirement compared with
Height is not suitable for industrial purification.Therefore it researches and develops new purification technique and is used for industry, to obtain higher general flavone quality point
Several guava leaf extract products is particularly significant.
Invention content
The technical problems to be solved by the invention are to overcome the shortcomings of to mention in background above technology and defect, provide one
The method of purification of Flavonoid substances Quercetin in kind Guava Leaf.
In order to solve the above technical problems, technical solution proposed by the present invention is provides Flavonoid substances in a kind of Guava Leaf
The method of purification of Quercetin, includes the following steps:
(1) by Guava Leaf grinding and sieving, water is added in obtained Guava Leaf powder, is heated, filtering obtains a kind stone
Pomegranate leaf extract flavones crude product;
(2) the flavones crude product obtained after the step (1) is dissolved in ethanol solution, supernatant is taken to import after filtering super
It filters and ultrafiltration cycle is carried out in device, obtain ultrafiltration membrance filter solution;
(3) it is dissolved in acetonitrile using Quercetin monomer as template molecule, adds in function monomer 4- vinyl and adjoin throat after room
Temperature is lower to be stirred, and template molecule is made fully to be acted on function monomer, crosslinking agent and initiator is then added in, is surpassed after abundant mixing
Sound is passed through nitrogen and forms oxygen-free environment, sealing, which is placed in thermostatical oil bath, carries out thermal polymerization, polymerize to remove the oxygen of dissolving
Product is eluted with methanol-acetic acid solution in Soxhlet extractor, and to remove template molecule, again with methanol washes away excessive acetic acid,
It is dried in vacuo in vacuum drying oven, the molecularly imprinted polymer after being synthesized;
(4) molecularly imprinted polymer obtained after the step (3) synthesis is accurately weighed, is obtained after adding in the step (2)
The ultrafiltration membrance filter solution arrived, carries out closed concussion in constant-temperature table, obtains being adsorbed with the molecularly imprinted polymer of Quercetin,
The Flavonoid substances Quercetin is obtained after centrifugation, the molecularly imprinted polymer after centrifugation may be reused.
Above-mentioned method of purification, it is preferred that in the step (1), be sieved for 40-60 mesh sieve, the water volume of addition is
38-42 times of Guava Leaf powder product.
Preferably, in the step (1), heating temperature is 50-70 DEG C, heating time 25-30min.
Preferably, in the step (2), the film that ultrafiltration cycle uses is retains the film that relative molecular mass is 10000.
Preferably, in the step (2), the time of ultrafiltration cycle is 2-3h, pressure 0.3-0.9MPa, temperature 30-
60℃。
Preferably, in the step (3), acetonitrile is no less than 40mL, and mixing time is no less than 4h, ultrasonic time 10-
20min is passed through the nitrogen time as 20-30min.
Preferably, in the step (3), crosslinking agent is ethylene glycol dimethacrylate (EDMA), and initiator is azo
Bis-isobutyronitrile (AIBN), the dosage of the initiator is function monomer and the 1-2% of crosslinking agent gross mass.
Preferably, in the step (3), oil bath pot temperature be 50-70 DEG C, the thermal polymerization time be 22-26h, methanol-acetic acid
The volume ratio of solution is methanol: acetic acid=9: 1, elution time 46-50h, and vacuum drying temperature is 30-50 DEG C, vacuum drying
Time is 22-26h.
Preferably, in the step (4), ultrafiltration membrance filter solution 10mL is added in every 30mg molecularly imprinted polymers.
Preferably, in the step (4), the temperature of constant-temperature table is 30-50 DEG C, and the concussion time is 22-26h.
Membrane separation technique and molecular imprinting technology are the purification new technologies researched and developed in recent years.Membrane separation technique has normal
The advantages that temperature operation, high selectivity, adaptable, simple for process, pollution are lacked.Molecular imprinting technology be according to antibody and antigen,
Enzyme-to-substrate etc. is principle there are the Molecular Recognization of nature, has identification target point using target molecule as templated synthesis
A kind of technology of the macromolecule imprinted polymer of son.It is of the invention by UF membrane since the impurity contained in flavones crude product is more
Technology is combined with molecular imprinting technology, and Guava Leaf crude extract is detached using membrane separation technique, is separated
Feed liquid further refines purification with the imprinted polymer of synthesis again, obtains the Guava Leaf Flavonoid substances Quercetin of purifying, from
Molecularly imprinted polymer after the heart may be reused.
Compared with prior art, beneficial effects of the present invention are:
1st, method of purification of the invention, membrane separation technique is combined with molecular imprinting technology, using membrane separation technique pair
Guava Leaf crude extract is detached, and the feed liquid separated further refines purification with the imprinted polymer of synthesis again, obtains
The Guava Leaf Flavonoid substances Quercetin that must be purified, environmental protection simple for process, income at low cost is high, and refining effect is good.
2nd, method of purification of the invention, membrane separation technique have normal-temperature operation, high selectivity, it is adaptable, simple for process,
The advantages that pollution is few, the accuracy of molecular imprinting technology identification target molecule is high, and high selectivity is pollution-free, and reliability is high, and
The molecularly imprinted polymer obtained after centrifugation may be reused, and be effectively saved process costs.
Specific embodiment
It is of the invention for the ease of understanding, the present invention is done below in conjunction with preferred embodiment and more comprehensively, is meticulously described,
But protection scope of the present invention is not limited to specific examples below.
Unless otherwise defined, all technical terms used hereinafter are generally understood meaning phase with those skilled in the art
Together.Technical term used herein is intended merely to the purpose of description specific embodiment, is not intended to the limitation present invention's
Protection domain.
Unless otherwise specified, various raw material, reagent, the instrument and equipment etc. used in the present invention can pass through city
Field is commercially available or can be prepared by existing method.
Embodiment 1:
The method of purification of Flavonoid substances Quercetin, includes the following steps in a kind of Guava Leaf of the present invention:
(1) Guava Leaf is smashed it through into 40 mesh sieve, water is added in obtained Guava Leaf powder, the water volume of addition is
40 times of Guava Leaf powder product, are heated to 60 DEG C and maintain 25min, filter, obtain guava leaf extract flavones crude product;
(2) the flavones crude product obtained after the step (1) is dissolved in ethanol solution, supernatant is taken to import after filtering super
It filters and ultrafiltration cycle 2h is carried out in device, film of the film used to retain relative molecular mass 10000, operating pressure 0.3MPa,
The temperature of supernatant is 30 DEG C, obtains ultrafiltration membrance filter solution;
(3) it is dissolved in Quercetin monomer as template molecule in acetonitrile (no less than 40mL), adds in function monomer 4- ethylene
Base adjoins throat and is no less than 4h after stirring at room temperature, and template molecule is made fully to be acted on function monomer, then adds in crosslinking agent and draws
Agent is sent out, crosslinking agent is ethylene glycol dimethacrylate, and initiator is azodiisobutyronitrile, and the dosage of the initiator is function
The 1.5% of monomer and crosslinking agent gross mass carries out ultrasound 10min after abundant mixing to remove the oxygen of dissolving, is passed through nitrogen
20min forms oxygen-free environment, and sealing is placed in 60 DEG C of thermostatical oil baths and carries out thermal polymerization for 24 hours, and polymerizate is with volume ratio
Methanol: acetic acid=9: 1 methanol-acetic acid solution elutes 48h in Soxhlet extractor, again with methanol washes away excessive acetic acid, in
It is dried in vacuo for 24 hours for 40 DEG C in vacuum drying oven, the molecularly imprinted polymer after being synthesized;
(4) the molecularly imprinted polymer 30mg obtained after the step (3) synthesis is accurately weighed, adds in the step (2)
The ultrafiltration membrance filter solution 10mL obtained afterwards carries out closed concussion in 40 DEG C of constant-temperature tables and obtains the Huang for 24 hours, after centrifugation
Letones Quercetin.
After testing, it is 64.69% to measure the rejection after the step (2), and the flavones mass fraction obtained after purification is
72% in ultrafiltration membrance filter solution, Quercetin maximum apparent adsorption quantity QmaxFor 3.675mg/g.
The test method of rejection:Protein content in solution is tested with spectrophotometry, calculates rejection, step is such as
Under:
1. the configuration of standard solution:Bovine serum albumin(BSA) is dried under vacuum to constant weight at 105 DEG C of temperature, accurately weighs ox blood
Pure albumen 1.000g is dissolved in 1000ml volumetric flasks, respectively draw bovine serum albumin solution 0.5,1.0,1.5,2.0,2.5,
3.0th, 3.5,4.0,4.5,5.0,6.0,7.0,8.0,9.0,10.0ml is in 10ml volumetric flasks plus distilled water is diluted to scale, matches
Be set to a concentration of 50,100,150,200,250,300,350,400,450,500,600,700,800,900, the ox of 1000mg/l
Seralbumin standard solution.
2. the making of standard curve:Protein has the ultraviolet light of 280nm absorption maximum, and protein solution 280nm absorbs
Value is directly proportional to its concentration.By 7.3.1 standard solution under wavelength 280nm, with 1cm cuvettes, on ultraviolet specrophotometer
Optical density is measured, distilled water is blank.Using protein concentration C as abscissa, absorbance A is charted for ordinate, makes standard song
Line.
3. the preparation of sample solution:The protein of a certain molecular weight is selected, is configured to a concentration of 1000mg/l -3000mg/
The protein solution of l uses after ovalbumin solution centrifugation or filtering, and the protein solution of preparation is commented as ultrafiltration film properties
The solution of valency uses.
4. the evaluation of ultrafiltration membrane:The sample solution being configured under 0.3MPa, normal temperature condition, is operated by ultrafiltration membrane,
Ultrafiltrate is collected after 20min, stoste measures optical density in 280nm ultraviolet regions respectively with ultrafiltrate, checked in from standard curve
Corresponding concentration.
5. the calculating of rejection:Rejection=(1-C1/C2) × 100%, wherein, C1For ultrafiltrate concentration, C2It is dense for stoste
Degree.
Maximum apparent adsorption quantity computational methods:
The bond type of absorption point position can be obtained, with reference to the equilibrium constant and Bmax etc. by Scatchard molecules
Important information.Scatchard equations are:
In formula, Q and QmaxFor balance binding capacity and maximum performance binding capacity;CeFor equilibrium concentration in solution;KDFor bound site
The equilibrium dissociation constant of point.It, can be in the hope of K according to linear relationship slope and intercept when being mapped with Q/C to QDAnd Qmax, two
Parameter.
Membrane separation technique is combined, using membrane separation technique pair by the method for purification of the present embodiment with molecular imprinting technology
Guava Leaf crude extract is detached, and the feed liquid separated further refines purification with the imprinted polymer of synthesis again, obtains
The Guava Leaf Flavonoid substances Quercetin that must be purified, environmental protection simple for process, income at low cost is high, and refining effect is good.
Embodiment 2:
The method of purification of Flavonoid substances Quercetin, includes the following steps in a kind of Guava Leaf of the present invention:
(1) Guava Leaf is smashed it through into 60 mesh sieve, water is added in obtained Guava Leaf powder, the water volume of addition is
40 times of Guava Leaf powder product, are heated to 60 DEG C and maintain 30min, filter, obtain guava leaf extract flavones crude product;
(2) the flavones crude product obtained after the step (1) is dissolved in ethanol solution, supernatant is taken to import after filtering super
It filters and ultrafiltration cycle 3h is carried out in device, film of the film used to retain relative molecular mass 10000, operating pressure 0.4MPa,
The temperature of supernatant is 50 DEG C, obtains ultrafiltration membrance filter solution;
(3) it is dissolved in Quercetin monomer as template molecule in acetonitrile (no less than 40mL), adds in function monomer 4- ethylene
Base adjoins throat and is no less than 4h after stirring at room temperature, and template molecule is made fully to be acted on function monomer, then adds in crosslinking agent and draws
Agent is sent out, crosslinking agent is ethylene glycol dimethacrylate, and initiator is azodiisobutyronitrile, and the dosage of the initiator is function
The 1.5% of monomer and crosslinking agent gross mass carries out ultrasound 20min after abundant mixing to remove the oxygen of dissolving, is passed through nitrogen
30min forms oxygen-free environment, and sealing is placed in 60 DEG C of thermostatical oil baths and carries out thermal polymerization for 24 hours, and polymerizate is with volume ratio
Methanol: acetic acid=9: 1 methanol-acetic acid solution elutes 48h in Soxhlet extractor, again with methanol washes away excessive acetic acid, in
It is dried in vacuo for 24 hours for 30 DEG C in vacuum drying oven, the molecularly imprinted polymer after being synthesized;Blank polymerization is prepared in the same way
Object;
(4) the molecularly imprinted polymer 30mg obtained after the step (3) synthesis is accurately weighed, adds in the step (2)
The ultrafiltration membrance filter solution 10mL obtained afterwards carries out closed concussion in 40 DEG C of constant-temperature tables and obtains the Huang for 24 hours, after centrifugation
Letones Quercetin.
After testing, it is 64.37% to measure the rejection after the step (2), and the flavones mass fraction obtained after purification is
71% in ultrafiltration membrance filter solution.Quercetin maximum apparent adsorption quantity QmaxFor 3.038mg/g.
The test method of rejection:Protein content in solution is tested with spectrophotometry, calculates rejection, step is such as
Under:
1. the configuration of standard solution:Bovine serum albumin(BSA) is dried under vacuum to constant weight at 105 DEG C of temperature, accurately weighs ox blood
Pure albumen 1.000g is dissolved in 1000ml volumetric flasks, respectively draw bovine serum albumin solution 0.5,1.0,1.5,2.0,2.5,
3.0th, 3.5,4.0,4.5,5.0,6.0,7.0,8.0,9.0,10.0ml is in 10ml volumetric flasks plus distilled water is diluted to scale, matches
Be set to a concentration of 50,100,150,200,250,300,350,400,450,500,600,700,800,900, the ox of 1000mg/l
Seralbumin standard solution.
2. the making of standard curve:Protein has the ultraviolet light of 280nm absorption maximum, and protein solution 280nm absorbs
Value is directly proportional to its concentration.By 7.3.1 standard solution under wavelength 280nm, with 1cm cuvettes, on ultraviolet specrophotometer
Optical density is measured, distilled water is blank.Using protein concentration C as abscissa, absorbance A is charted for ordinate, makes standard song
Line.
3. the preparation of sample solution:The protein of a certain molecular weight is selected, is configured to a concentration of 1000mg/l -3000mg/
The protein solution of l uses after ovalbumin solution centrifugation or filtering, and the protein solution of preparation is commented as ultrafiltration film properties
The solution of valency uses.
4. the evaluation of ultrafiltration membrane:The sample solution being configured under 0.3MPa, normal temperature condition, is operated by ultrafiltration membrane,
Ultrafiltrate is collected after 20min, stoste measures optical density in 280nm ultraviolet regions respectively with ultrafiltrate, checked in from standard curve
Corresponding concentration.
5. the calculating of rejection:Rejection=(1-C1/C2) × 100%, wherein, C1For ultrafiltrate concentration, C2It is dense for stoste
Degree.
Maximum apparent adsorption quantity computational methods:
The bond type of absorption point position can be obtained, with reference to the equilibrium constant and Bmax etc. by Scatchard molecules
Important information.Scatchard equations are:
In formula, Q and QmaxFor balance binding capacity and maximum performance binding capacity;CeFor equilibrium concentration in solution;KDFor bound site
The equilibrium dissociation constant of point.It, can be in the hope of K according to linear relationship slope and intercept when being mapped with Q/C to QDAnd QmaxTwo ginsengs
Number.
Membrane separation technique is combined, using membrane separation technique pair by the method for purification of the present embodiment with molecular imprinting technology
Guava Leaf crude extract is detached, and the feed liquid separated further refines purification with the imprinted polymer of synthesis again, obtains
The Guava Leaf Flavonoid substances Quercetin that must be purified, environmental protection simple for process, income at low cost is high, and refining effect is good.
Claims (10)
1. the method for purification of Flavonoid substances Quercetin, includes the following steps in a kind of Guava Leaf:
(1) by Guava Leaf grinding and sieving, water is added in obtained Guava Leaf powder, is heated, filtering obtains Guava Leaf
Extract flavone crude product;
(2) the flavones crude product obtained after the step (1) is dissolved in ethanol solution, supernatant is taken to import ultrafiltration dress after filtering
Middle progress ultrafiltration cycle is put, obtains ultrafiltration membrance filter solution;
(3) it is dissolved in acetonitrile using Quercetin monomer as template molecule, adds in function monomer 4- vinyl and adjoin throat after at room temperature
Stirring, makes template molecule fully be acted on function monomer, then adds in crosslinking agent and initiator, carried out after abundant mixing it is ultrasonic with
The oxygen of dissolving is removed, nitrogen is passed through and forms oxygen-free environment, sealing, which is placed in thermostatical oil bath, carries out thermal polymerization, polymerizate
It is eluted in Soxhlet extractor with methanol-acetic acid solution, to remove template molecule, again with methanol washes away excessive acetic acid, Yu Zhen
It is dried in vacuo in empty baking oven, the molecularly imprinted polymer after being synthesized;
(4) obtained molecularly imprinted polymer after the step (3) synthesis is accurately weighed, adds in after the step (2) what is obtained
Ultrafiltration membrance filter solution carries out closed concussion in constant-temperature table, obtains being adsorbed with the molecular engram of Flavonoid substances Quercetin
Polymer obtains the Flavonoid substances Quercetin after centrifugation.
2. method of purification according to claim 1, which is characterized in that in the step (1), be sieved for 40-60 mesh sieve,
The water volume of addition is 38-42 times of Guava Leaf powder product.
3. method of purification according to claim 1 or 2, which is characterized in that in the step (1), heating temperature 50-70
DEG C, heating time 25-30min.
4. method of purification according to claim 1, which is characterized in that in the step (2), the ultrafiltration film that uses of cycle for
Retain the film that relative molecular mass is 10000.
5. the method for purification according to claim 1 or 4, which is characterized in that in the step (2), the time of ultrafiltration cycle
For 2-3h, pressure 0.3-0.9MPa, temperature is 30-60 DEG C.
6. method of purification according to claim 1, which is characterized in that in the step (3), acetonitrile is no less than 40mL, stirs
Mixing the time is no less than 4h, ultrasonic time 10-20min, is passed through the nitrogen time as 20-30min.
7. the method for purification according to claim 1 or 6, which is characterized in that in the step (3), crosslinking agent is ethylene glycol
Dimethylacrylate, initiator are azodiisobutyronitrile, and the dosage of the initiator is function monomer and crosslinking agent gross mass
1-2%.
8. the method for purification according to claim 1 or 6, which is characterized in that in the step (3), oil bath pot temperature is 50-
70 DEG C, the thermal polymerization time is 22-26h, and the volume ratio of methanol-acetic acid solution is methanol: acetic acid=9: 1, elution time 46-
50h, vacuum drying temperature are 30-50 DEG C, vacuum drying time 22-26h.
9. method of purification according to claim 1, which is characterized in that in the step (4), per the polymerization of 30mg molecular engrams
Ultrafiltration membrance filter solution 10mL is added in object.
10. method of purification according to claim 1, which is characterized in that in the step (4), the temperature of constant-temperature table is
30-50 DEG C, the concussion time is 22-26h.
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110669172A (en) * | 2019-09-03 | 2020-01-10 | 湖州耕香生物科技有限公司 | Preparation method and application of molecularly imprinted polymer |
CN112513162A (en) * | 2018-07-05 | 2021-03-16 | Upl有限公司 | Novel compositions for bittering agents |
CN115433315A (en) * | 2022-10-18 | 2022-12-06 | 江西省科学院应用化学研究所 | Molecularly imprinted polymer, preparation method and application thereof, and method for simultaneously extracting three flavonoid glycoside compounds from polygonum multiflorum leaves |
CN115478026A (en) * | 2022-06-29 | 2022-12-16 | 中南大学 | Radioresistant coccus and application thereof |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050283851A1 (en) * | 2004-03-09 | 2005-12-22 | Sharon Abrahams | Novel genes encoding proteins involved in proanthocyanidin synthesis |
CN103055831A (en) * | 2011-10-20 | 2013-04-24 | 中国科学院兰州化学物理研究所 | Preparation method of inorganic core-shell type quercetin molecularly imprinted polymer microsphere |
CN106749145A (en) * | 2016-12-14 | 2017-05-31 | 佛山科学技术学院 | A kind of method that Quercetin is extracted from Guava Leaf |
-
2017
- 2017-12-28 CN CN201711466673.6A patent/CN108191804B/en not_active Expired - Fee Related
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050283851A1 (en) * | 2004-03-09 | 2005-12-22 | Sharon Abrahams | Novel genes encoding proteins involved in proanthocyanidin synthesis |
CN103055831A (en) * | 2011-10-20 | 2013-04-24 | 中国科学院兰州化学物理研究所 | Preparation method of inorganic core-shell type quercetin molecularly imprinted polymer microsphere |
CN106749145A (en) * | 2016-12-14 | 2017-05-31 | 佛山科学技术学院 | A kind of method that Quercetin is extracted from Guava Leaf |
Non-Patent Citations (2)
Title |
---|
VUSUMZI PAKADE ET AL.: "Molecular imprinted polymer for solid-phase extraction of flavonol aglycones from Moringa oleifera extracts", 《J. SEP. SCI.》 * |
朱俊访 等: "分子印迹聚合物在黄酮类化合物分离中的应用进展", 《中成药》 * |
Cited By (7)
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---|---|---|---|---|
CN112513162A (en) * | 2018-07-05 | 2021-03-16 | Upl有限公司 | Novel compositions for bittering agents |
EP3818102A4 (en) * | 2018-07-05 | 2022-03-30 | UPL Ltd | Novel compositions for bitterants |
CN110669172A (en) * | 2019-09-03 | 2020-01-10 | 湖州耕香生物科技有限公司 | Preparation method and application of molecularly imprinted polymer |
CN115478026A (en) * | 2022-06-29 | 2022-12-16 | 中南大学 | Radioresistant coccus and application thereof |
CN115478026B (en) * | 2022-06-29 | 2023-07-04 | 中南大学 | Radioresistant coccus and application thereof |
CN115433315A (en) * | 2022-10-18 | 2022-12-06 | 江西省科学院应用化学研究所 | Molecularly imprinted polymer, preparation method and application thereof, and method for simultaneously extracting three flavonoid glycoside compounds from polygonum multiflorum leaves |
CN115433315B (en) * | 2022-10-18 | 2023-12-22 | 江西省科学院应用化学研究所 | Molecularly imprinted polymer, preparation method and application thereof, and method for simultaneously extracting three flavonoid glycoside compounds from polygonum multiflorum leaves |
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