CN106045958B - A method of separating-purifying myricetin and dihydromyricetin from vine tea - Google Patents
A method of separating-purifying myricetin and dihydromyricetin from vine tea Download PDFInfo
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- CN106045958B CN106045958B CN201610621955.8A CN201610621955A CN106045958B CN 106045958 B CN106045958 B CN 106045958B CN 201610621955 A CN201610621955 A CN 201610621955A CN 106045958 B CN106045958 B CN 106045958B
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- KJXSIXMJHKAJOD-LSDHHAIUSA-N (+)-dihydromyricetin Chemical compound C1([C@@H]2[C@H](C(C3=C(O)C=C(O)C=C3O2)=O)O)=CC(O)=C(O)C(O)=C1 KJXSIXMJHKAJOD-LSDHHAIUSA-N 0.000 title claims abstract description 122
- KQILIWXGGKGKNX-UHFFFAOYSA-N dihydromyricetin Natural products OC1C(=C(Oc2cc(O)cc(O)c12)c3cc(O)c(O)c(O)c3)O KQILIWXGGKGKNX-UHFFFAOYSA-N 0.000 title claims abstract description 62
- IKMDFBPHZNJCSN-UHFFFAOYSA-N Myricetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC(O)=C(O)C(O)=C1 IKMDFBPHZNJCSN-UHFFFAOYSA-N 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 49
- 229940116852 myricetin Drugs 0.000 title claims abstract description 49
- PCOBUQBNVYZTBU-UHFFFAOYSA-N myricetin Natural products OC1=C(O)C(O)=CC(C=2OC3=CC(O)=C(O)C(O)=C3C(=O)C=2)=C1 PCOBUQBNVYZTBU-UHFFFAOYSA-N 0.000 title claims abstract description 49
- 235000007743 myricetin Nutrition 0.000 title claims abstract description 49
- 241001122767 Theaceae Species 0.000 title claims abstract description 42
- 239000006228 supernatant Substances 0.000 claims abstract description 89
- 238000001556 precipitation Methods 0.000 claims abstract description 69
- 239000000284 extract Substances 0.000 claims abstract description 64
- 238000000703 high-speed centrifugation Methods 0.000 claims abstract description 55
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- 238000000605 extraction Methods 0.000 claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000000843 powder Substances 0.000 claims abstract description 29
- 238000001291 vacuum drying Methods 0.000 claims abstract description 19
- 239000000706 filtrate Substances 0.000 claims abstract description 17
- 239000007788 liquid Substances 0.000 claims abstract description 15
- 230000010355 oscillation Effects 0.000 claims abstract description 12
- 238000001914 filtration Methods 0.000 claims abstract description 9
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 135
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 28
- 239000013078 crystal Substances 0.000 claims description 26
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 14
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 14
- 238000001035 drying Methods 0.000 claims description 11
- 238000000746 purification Methods 0.000 claims description 11
- 238000010992 reflux Methods 0.000 claims description 11
- 239000002244 precipitate Substances 0.000 claims description 10
- 229920002678 cellulose Polymers 0.000 claims description 9
- 239000001913 cellulose Substances 0.000 claims description 9
- 238000005119 centrifugation Methods 0.000 claims description 7
- 238000002425 crystallisation Methods 0.000 claims description 7
- 230000008025 crystallization Effects 0.000 claims description 7
- 238000000151 deposition Methods 0.000 claims description 7
- 230000008021 deposition Effects 0.000 claims description 7
- 238000000926 separation method Methods 0.000 claims description 7
- 244000132436 Myrica rubra Species 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims description 5
- 239000004327 boric acid Substances 0.000 claims description 5
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims description 5
- 244000144992 flock Species 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 238000013019 agitation Methods 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims 1
- 150000003839 salts Chemical class 0.000 claims 1
- 239000013049 sediment Substances 0.000 claims 1
- 239000002904 solvent Substances 0.000 abstract description 4
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
- 230000003000 nontoxic effect Effects 0.000 abstract description 3
- 238000011084 recovery Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 235000019439 ethyl acetate Nutrition 0.000 description 41
- 235000019441 ethanol Nutrition 0.000 description 21
- 239000000243 solution Substances 0.000 description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Substances OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 230000001476 alcoholic effect Effects 0.000 description 6
- 238000002955 isolation Methods 0.000 description 6
- 230000020477 pH reduction Effects 0.000 description 6
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000004064 recycling Methods 0.000 description 4
- 238000005057 refrigeration Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 241000219000 Populus Species 0.000 description 3
- 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 3
- 238000004090 dissolution Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 229930003944 flavone Natural products 0.000 description 3
- 235000011949 flavones Nutrition 0.000 description 3
- 238000002386 leaching Methods 0.000 description 3
- 238000005498 polishing Methods 0.000 description 3
- 239000013558 reference substance Substances 0.000 description 3
- 208000006379 syphilis Diseases 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 235000009388 Parthenocissus quinquefolia Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 150000002212 flavone derivatives Chemical class 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 241000563984 Ampelopsis Species 0.000 description 1
- 241000345998 Calamus manan Species 0.000 description 1
- 206010017553 Furuncle Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 241001018563 Nekemias grossedentata Species 0.000 description 1
- 241000219099 Parthenocissus quinquefolia Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 206010000269 abscess Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 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
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005485 electric heating Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- -1 flavone compound Chemical class 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000000874 microwave-assisted extraction Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 235000012950 rattan cane Nutrition 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
- 230000001568 sexual effect Effects 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
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
- C07D311/32—2,3-Dihydro derivatives, e.g. flavanones
-
- 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
Abstract
The method of the invention discloses a kind of from vine tea separating-purifying myricetin and dihydromyricetin, this method comprises the following steps：Vine tea is weighed, it is selected, it crushes, crosses 35 45 mesh sieve；Alcohol is added, impregnates 6 10 hours, is then refluxed for extraction 35 hours；Filtrate is obtained by filtration in extraction liquid；Recovered alcohol to filtrate alcohol content is 15% hereinafter, being then concentrated in vacuo to obtain the medicinal extract of proportion 1.0 1.5 from filtrate；Concentrated extract is sufficiently stirred with hot water and is used ultrasonic oscillation, high speed centrifugation collect supernatant；The supernatant refrigerated overnight that will be obtained, high speed centrifugation collect precipitation, and vacuum drying obtains yellow greenish powder primary extract；It isolates and purifies；Obtain 98% or more high-purity myricetin and dihydromyricetin sterling.The method of the present invention equipment is simple, simple for process feasible, is convenient for industrialized production；Solvent for use nontoxic residue-free, not generation environment pollution；Utilization rate that the present invention isolates and purifies is high, the rate of recovery is high, and gained myricetin, dihydromyricetin purity are high.
Description
Technical field
The present invention relates to technical field of Chinese medicine, are more particularly to one kind separating-purifying myricetin and dihydro poplar from vine tea
The method of syphilis.
Background technology
Vine tea category Vitaceae, Ampelopsis, scientific name are ampelopsis grossdentata (Ampelopsis grossedentata), taste
It is sweet, light, it is cool in nature, there is clearing heat and detoxicating, wind-damp dispelling, strengthening the bones and muscles, anti-inflammatory, analgesia and other effects.It is civil that guarantor is made in its young young stem and leaf
Strong tea, has the history of centuries for treating the diseases such as cold, fever, abscess of throat, icteric hepatitis, blister furuncle, is a kind of typical case
Medical and edible dual purpose plant.Vine tea principle active component is flavone compound, and with dihydromyricetin
(Dihydromyricetin, abbreviation DMY) content highest.Studies have shown that general flavone content is 43.4%~44.0% in vine tea
(mass fraction), dihydromyricetin cellulose content are up to 37.4%~40%, but related with the factors such as geography, climatic environment.In recent years,
Existing some scholars have carried out the research that dihydromyricetin is extracted from plant both at home and abroad, are typically extraction with organic solvent
Agent, such as ethyl acetate, n-butanol, ethyl alcohol.This method extraction cost is high, the high energy consumption of recycling design, there is also it is time-consuming, carry
In place of the deficiencies of taking low rate, poor reproducibility.And solvent for use is usually toxic, and it is dangerous, easily environment and operating personnel are caused
Harm.Though supercritical extract dihydromyricetin has many advantages, such as to save reagent, pollution-free, the rate of recovery is poor, super in order to obtain
Critical condition, equipment one-time investment is larger, and operating cost is high.In addition, also macroreticular resin absorbing method, microwave extraction method, inverse
Stream method, ethyl alcohol auxiliary law and ultrasonic wave secondary solvent extraction method.
Invention content
(1) technical problems to be solved
The technical problem to be solved in the present invention is to how low cost, low energy consumption, safe and nontoxic, free of contamination extraction rattan
Active ingredient in tea, and a kind of method of separating-purifying myricetin and dihydromyricetin from vine tea is provided
(2) technical solution
In order to solve the above technical problem, the present invention provides one kind separating-purifying myricetin and dihydromyricetins from vine tea
The method (raw materials used acquisition purchased in market) of element, this method comprises the following steps：
Step 1：Preparation：Vine tea is weighed, it is selected, it crushes, crosses 35-45 mesh sieve；Too thin, too coarse extraction is inefficient；
Step 2：Extraction：The alcohol of the 75-85% of 18-22 times of vine tea weight is added, is adjusted with 5% sodium bicarbonate solution
PH value is 7.0-8.0, is impregnated 6-10 hours, then reflux extracts 3-5 hours at a temperature of 35-45 DEG C；Alkaline condition is conducive to
Extraction；
Step 3：Secondary extraction：The alcohol of the 75-85% of 8-12 times of vine tea weight is added, flows back at a temperature of 35-45 DEG C
Extraction 3-5 hours；
Step 4：Filtering：Merging extracts liquid twice, and filtrate is obtained by filtration；Filter out impurity；
Step 5：Recovered alcohol：Recovered alcohol to alcohol content is 15% hereinafter, reach alcohol-free taste from filtrate,
Then it is concentrated in vacuo to obtain the medicinal extract of proportion 1.0-1.5；
Step 6：Water is heavy：Concentrated extract is sufficiently stirred with 90-98 DEG C of hot water of 18-22 times of weight and uses ultrasonic wave
Concussion 2.5-3.5 hours, high speed centrifugation at a temperature of 80-90 DEG C collect supernatant；
Step 7：Centrifugation：The supernatant refrigerated overnight that step 6 is obtained carries out high speed centrifugation in second day, collects precipitation,
Vacuum drying obtains yellow greenish powder primary extract；Refrigeration is conducive to myricetin and dihydromyricetin crystalline deposit, ensures target overnight
Object sufficient crystallising is precipitated；
Step 8：It isolates and purifies；The primary extract that step 7 obtains is isolated and purified to obtain 98% or more high-purity red bayberry
Element and dihydromyricetin sterling.
Preferably, in step 8, the purification procedures are as follows：
1) yellow greenish powder primary extract will be obtained in step 7, yellowish green toner is added in the ethyl acetate of point 3 use, 9 times of weight
In last primary extract, uniform stirring and with ultrasonic vibration 2.5-3.5 hours, ethyl acetate layer becomes yellow green, and merging obtains three times
Ethyl acetate supernatant and the white depositions that obtain three times；
2) 8-12 times of 5% sodium bicarbonate solution of white depositions that step 1) obtains is dissolved, high speed centrifugation takes supernatant
Supernatant is acidified by liquid, refrigerated overnight, obtains white crystals precipitation, and drying obtains the dihydromyricetin of purity at least 90%；Again
90 DEG C or more the hot water that white crystals are precipitated to 5 times of weight are sufficiently stirred dissolving, high speed centrifugation, and precipitation and separation collects supernatant
Liquid, by supernatant refrigerated overnight, high speed centrifugation, collection obtains white, needle-shaped crystals, with pure water 3 times, obtains white needles
Crystallization, drying obtain the white needles dihydromyricetin cellulose crystal of 98% or more purity；
3) ethyl acetate is recycled in the supernatant obtained from step 1), vacuum drying obtains yellow powder, by yellow powder
It is dissolved using 5% sodium bicarbonate solution, high speed centrifugation, removes precipitation and collect supernatant, supernatant is acidified, is chilled overnight, point
The needle-shaped myricetin crystallization of 98% or more yellow green can be obtained from precipitating and being dried in vacuo.
Preferably, in step 8, the purification procedures are as follows：
1) yellow greenish powder primary extract will be obtained in step 7,5% boric acid solution of 10 times of primary extract weight is added, fully
Simultaneously ultrasonic vibration 2.5-3.5 hours are stirred, overnight precipitation is then allowed to stand；High speed centrifugation removes precipitation, collects supernatant；
2) 5% calcium hydroxide solution is added in supernatant, is slowly stirred, constantly generates white flock precipitate, adjusts steady
Determine until PH 7.5, then slowly more than half an hour, high speed centrifugation removes precipitation, collection supernatant for agitation；
3) supernatant being collected into hydrochloric acid is adjusted into pH value to 4.0, refrigerated overnight, high speed centrifugation precipitates, and collection obtains
The only precipitation of the mixture of myricetin and dihydromyricetin；
4) it is added three times the ethyl acetate of 9 times of weight in the precipitation that step 3) obtains, is sufficiently stirred simultaneously ultrasonic oscillation
It 3 hours or more, stands overnight；High speed centrifugation collects precipitation and supernatant respectively；
5) the white needles dihydromyricetin cellulose crystal for precipitating vacuum drying to get purity 98% for obtaining step 4)；
6) vacuum recycles ethyl acetate in the supernatant obtained from step 4), obtains body of paste, is added 10 times 95 DEG C or more
Hot water, fully dissolving, are filtered, cooling, and overnight, high speed centrifugation is collected and obtains the yellow needles myricetin crystalline substance of 98% or more purity
Body.
Preferably, in step 1, the sieve is 40 mesh.
Preferably, in step 2 and step 3, the extraction temperature is 40 DEG C.
Preferably, in step 5, the proportion is 1.15-1.20.
Preferably, in step 6, the heavy temperature of the water is 95 DEG C.
Preferably, in step 3), vacuum degree is -0.08--0.09MPa.
Preferably, described method includes following steps：
Step 1：Preparation：Vine tea is weighed, it is selected, it crushes, crosses 40 mesh sieve；
Step 2：Extraction：The alcohol of the weight concentration 80% of 20 times of vine tea weight is added, with 5% sodium bicarbonate solution tune
It is 7.5 to save pH value, is impregnated 8 hours, then reflux extracts 4 hours at a temperature of 40 DEG C；
Step 3：Secondary extraction：The alcohol of the weight concentration 80% of 10 times of vine tea weight is added, flows back at a temperature of 40 DEG C
Extraction 4 hours；
Step 4：Filtering：Merging extracts liquid twice, and filtrate is obtained by filtration；
Step 5：Recovered alcohol：Recovered alcohol to alcohol concentration is 15% hereinafter, being then concentrated in vacuo to from filtrate
To the medicinal extract of proportion 1.2；
Step 6：Water is heavy：95 DEG C of hot water of 20 times of weight of concentrated extract is sufficiently stirred and uses ultrasonic oscillation 3 small
When, high speed centrifugation at a temperature of 85 DEG C collects supernatant；
Step 7：Centrifugation：The supernatant refrigerated overnight that step 6 is obtained carries out high speed centrifugation in second day, collects precipitation,
Vacuum drying obtains yellow greenish powder primary extract；
Step 8：It isolates and purifies；The primary extract that step 7 obtains is isolated and purified to obtain 98% or more high-purity red bayberry
Element and dihydromyricetin sterling.
The present invention also provides above-mentioned from vine tea obtained by the method for separating-purifying myricetin and dihydromyricetin
Myricetin and dihydromyricetin.
(3) advantageous effect
The used equipment of the method for the present invention is simple, simple for process feasible, is convenient for industrialized production；Solvent for use is nontoxic without residual
It stays, generation environment does not pollute, and the organic solvent usage amount of final wash crystallization is small；The present invention utilizes vine tea extraction separation and purification
Utilization rate is high, the gained rate of recovery is high, all 90% or more, gained myricetin, dihydromyricetin purity are also very high, can directly use
In pharmaceutical industries.
Specific implementation mode
Embodiments of the present invention are described in further detail with reference to embodiment.Following embodiment is for illustrating this
Invention, but cannot be used for limiting the scope of the invention.
Used solvent, HCL, sodium bicarbonate, ethyl acetate are that food grade or analysis are pure below, must not use work
Industry grade.
Embodiment 1
1) 1000 kilograms, the selected removal of impurities of vine tea, vine tea crushes, and crosses 40 mesh sieve, but should not be meticulous, 1000 kilograms of polishing of being weighed
Total amount；
2) it extracts for the first time：20 ton of 80% alcoholic solution adjusts PH to 7.5 with sodium bicarbonate, and 10 tons of extractors are added and impregnate
Reflux extraction 4 hours at a temperature of 8 hours and then 40 DEG C；
3) it extracts for second：Reflux extraction 4 hours at a temperature of 10 ton of 80% 40 DEG C of alcoholic solution；
4) it is filtered for the first time with plate and frame type filter-press with second of leaching liquor, obtains totally 28.6 tons of the filtrate of merging；
5) recovered alcohol and concentration：Then recovered alcohol after filtrate merges is concentrated in vacuo to until alcohol concentration is less than 15%
The medicinal extract of proportion 1.2 is obtained, medicinal extract weight is 1340 kilograms；
6) medicinal extract is sufficiently stirred with 27 tons 95 DEG C or more of hot water and uses ultrasonic oscillation 3 hours, high under (85 DEG C) while hot
25.9 tons of supernatant is collected by centrifugation in speed；
7) supernatant enters freezer, and refrigerated overnight, high speed centrifugation collects precipitation within second day, and vacuum drying obtains yellow greenish powder
632 kilograms of primary extract；
8) 632 kilograms of yellow green primary extracts are isolated and purified with the myricetin and dihydromyricetin of 98% or more high-purity of extraction
Sterling；
8.1) the first isolation and purification method；
8.1.1 it is (every to analyze pure ethyl acetates point 3 times with 2700 kilograms for) 312 kilograms of yellow green primary extract obtained by step 7)
Secondary 900 kilograms) amount is added in yellow greenish powder, and it constantly mixes slowly and with ultrasonic vibration 3 hours, ethyl acetate layer became yellowish green
Color, 3 times of ethyl acetate layers of last time are basically unchanged color；
8.1.2 (1) 210 kilogram of white depositions) are respectively obtained and ethyl acetate supernatant combined wt 2800 is public three times
Jin；
8.1.3 dissolving, high speed centrifugation, supernatant acidification refrigeration) are easy to 1000 kilogram of 5% sodium bicarbonate of precipitation (1)
Overnight white crystals precipitation dry at least 129 kilogram 90% purity dihydromyricetin, white, needle-shaped crystals are with 600 kilograms
95 DEG C or more hot water are sufficiently stirred dissolving, and the precipitation that is centrifuged at a high speed collects supernatant, and supernatant refrigerated overnight high speed centrifugation is received
Collect white, needle-shaped crystals, after pure water 3 times yellow needles drying can obtain 98% or more white needles dihydromyricetin
123.3 kilograms of cellulose crystal.
8.1.4) 2800 kilograms of ethyl acetate supernatant；Supernatant recycles ethyl acetate, and vacuum drying obtains yellow powder 9.1
Kilogram, dissolving high speed centrifugation removing precipitation is easy using 100 kilogram of 5% sodium bicarbonate to yellow powder and collects supernatant 106.4
Kilogram, supernatant acidification is chilled overnight precipitation and separation and vacuum drying can obtain the needle-shaped myricetin of 98% or more yellow green and crystallize 8.7
Kilogram；
8.2) second of isolation and purification method；
8.2.1) 316 kilograms of yellow greenish powder primary extract obtained by step 7), it is molten to be added 3000 kilogram weights of 5% boric acid
Liquid is sufficiently stirred simultaneously ultrasonic vibration 3 hours, is then allowed to stand overnight precipitation；
8.2.2) rear high speed centrifugation removing precipitates to obtain 3260 kilograms of supernatant overnight；
8.2.3) 5% calcium hydroxide solution is added in supernatant, is slowly stirred, and constantly generates white flock precipitate, adjusts steady
Fixed slowly to be stirred more than half an hour until PH7.5, high speed centrifugation removes precipitation, collects 2980 kilograms of supernatant；
8.2.4 the supernatant) being collected into adjusts PH to 4.0, refrigerated overnight with hydrochloric acid；
8.2.5) rear high speed centrifugation precipitation, collection obtain the precipitation of the mixture of only myricetin and dihydromyricetin overnight
(2) 322 kilograms；
8.2.6) precipitation (2) detach myricetin and dihydromyricetin with 2700 kilograms of analysis pure ethyl acetate, and 900
Kilogram primary analysis pure ethyl acetate point is sufficiently stirred for 3 times and ultrasonic oscillation 3 hours or more, and high speed centrifugation precipitation collects supernatant
Liquid, precipitation with 900 kilograms of ethyl acetate ultrasounds and stirs be centrifuged at a high speed again precipitation and supernatant again, for the third time with 900
Kilogram ethyl acetate is stripped and centrifuges to precipitation；
8.2.7 the dihydromyricetin 121.7 of white needle-like crystals is obtained after the crystallization drying of (3) 119 kg whites) must be precipitated
Kilogram, 2690 kilograms of the supernatant three times of merging；
8.2.8) vacuum recycling ethyl acetate obtains 24.5 kilograms of body of paste, and 250 kilograms of 95 DEG C or more hot water are added in body of paste
Abundant dissolution filter, it is cold go to stay overnight high speed centrifugation collect yellow needles body and vacuum drying and obtain 8.5 kilograms of myricetin purity
98% or more.
Embodiment 2
1) 1000 kilograms, the selected removal of impurities of vine tea, vine tea crushes, and crosses 40 mesh sieve, but should not be meticulous, 1000 kilograms of polishing of being weighed
Total amount；
2) it extracts for the first time：20 ton of 80% alcoholic solution adjusts PH to 7.5 with sodium bicarbonate, and 10 tons of extractors are added and impregnate
Reflux extraction 4 hours at a temperature of 8 hours and then 40 DEG C；
3) it extracts for second：Reflux extraction 4 hours at a temperature of 10 ton of 80% 40 DEG C of alcoholic solution；
4) it is filtered for the first time with plate and frame type filter-press with second of leaching liquor, obtains totally 28.4 tons of the filtrate of merging；
5) recovered alcohol and concentration：Then recovered alcohol after filtrate merges is concentrated in vacuo to until alcohol concentration is less than 15%
The medicinal extract of proportion 1.2 is obtained, medicinal extract weight is 1290 kilograms；
6) medicinal extract is sufficiently stirred with 26 tons 95 DEG C or more of hot water and uses ultrasonic oscillation 3 hours, high under (85 DEG C) while hot
25.1 tons of supernatant is collected by centrifugation in speed；
7) supernatant enters freezer, and refrigerated overnight, high speed centrifugation collects precipitation within second day, and vacuum drying obtains yellow greenish powder
628 kilograms of primary extract；
8) 628 kilograms of yellow green primary extracts are isolated and purified with the myricetin and dihydromyricetin of 98% or more high-purity of extraction
Sterling；
8.1) the first isolation and purification method；
8.1.1 it is (every to analyze pure ethyl acetates point 3 times with 2700 kilograms for) 314 kilograms of yellow green primary extract obtained by step 7)
Secondary 900 kilograms) amount is added in yellow greenish powder, and it constantly mixes slowly and with ultrasonic vibration 3 hours, ethyl acetate layer became yellowish green
Color, 3 times of ethyl acetate layers of last time are basically unchanged color；
8.1.2 (1) 201 kilogram of white depositions) are respectively obtained and ethyl acetate supernatant combined wt 2880 is public three times
Jin；
8.1.3 dissolving, high speed centrifugation, supernatant acidification refrigeration) are easy to 1000 kilogram of 5% sodium bicarbonate of precipitation (1)
Overnight white crystals precipitation dry at least 132.7 kilogram 90% purity dihydromyricetin, white, needle-shaped crystals are public with 650
95 DEG C of jin or more hot water is sufficiently stirred dissolving, and the precipitation that is centrifuged at a high speed collects supernatant, supernatant refrigerated overnight high speed centrifugation
Collect white, needle-shaped crystals, after pure water 3 times yellow needles drying can obtain 98% or more white needles dihydro poplar
120.2 kilograms of syphilis crystal.
8.1.4) 2880 kilograms of ethyl acetate supernatant；Supernatant recycles ethyl acetate, and vacuum drying obtains yellow powder 9.2
Kilogram, dissolving high speed centrifugation removing precipitation is easy using 100 kilogram of 5% sodium bicarbonate to yellow powder and collects supernatant 110.1
Kilogram, supernatant acidification is chilled overnight precipitation and separation and vacuum drying can obtain the needle-shaped myricetin of 98% or more yellow green and crystallize 8.6
Kilogram；
8.2) second of isolation and purification method；
8.2.1) 314 kilograms of yellow greenish powder primary extract obtained by step 7), it is molten to be added 3000 kilogram weights of 5% boric acid
Liquid is sufficiently stirred simultaneously ultrasonic vibration 3 hours, is then allowed to stand overnight precipitation；
8.2.2) rear high speed centrifugation removing precipitates to obtain 3310 kilograms of supernatant overnight；
8.2.3) 5% calcium hydroxide solution is added in supernatant, is slowly stirred, and constantly generates white flock precipitate, adjusts steady
Fixed slowly to be stirred more than half an hour until PH7.5, high speed centrifugation removes precipitation, collects 3130 kilograms of supernatant；
8.2.4 the supernatant) being collected into adjusts PH to 4.0, refrigerated overnight with hydrochloric acid；
8.2.5) rear high speed centrifugation precipitation, collection obtain the precipitation of the mixture of only myricetin and dihydromyricetin overnight
(2) 317 kilograms；
8.2.6) precipitation (2) detach myricetin and dihydromyricetin with 2700 kilograms of analysis pure ethyl acetate, and 900
Kilogram primary analysis pure ethyl acetate point is sufficiently stirred for 3 times and ultrasonic oscillation 3 hours or more, and high speed centrifugation precipitation collects supernatant
Liquid, precipitation with 900 kilograms of ethyl acetate ultrasounds and stirs be centrifuged at a high speed again precipitation and supernatant again, for the third time with 900
Kilogram ethyl acetate is stripped and centrifuges to precipitation；
8.2.7 the dihydromyricetin of white needle-like crystals is obtained after the crystallization drying of (3) 129.6 kg whites) must be precipitated
119.8 kilograms, 2700 kilograms of the supernatant three times of merging；
8.2.8) vacuum recycling ethyl acetate obtains 25.5 kilograms of body of paste, and 250 kilograms of 95 degree or more hot water are added in body of paste
Abundant dissolution filter, it is cold go to stay overnight high speed centrifugation collect yellow needles body and vacuum drying and obtain 8.3 kilograms of myricetin purity
98% or more.
Embodiment 3
1) 1000 kilograms, the selected removal of impurities of vine tea, vine tea crushes, and crosses 40 mesh sieve, but should not be meticulous, 1000 kilograms of polishing of being weighed
Total amount；
2) it extracts for the first time：20 ton of 80% alcoholic solution adjusts PH to 7.5 with sodium bicarbonate, and 10 tons of extractors are added and impregnate
Reflux extraction 4 hours at a temperature of 8 hours and then 40 DEG C；
3) it extracts for second：Reflux extraction 4 hours at a temperature of 10 ton of 80% 40 DEG C of alcoholic solution；
4) it is filtered for the first time with plate and frame type filter-press with second of leaching liquor, obtains totally 29.1 tons of the filtrate of merging；
5) recovered alcohol and concentration：Then recovered alcohol after filtrate merges is concentrated in vacuo to until alcohol concentration is less than 15%
The medicinal extract of proportion 1.2 is obtained, medicinal extract weight is 1295 kilograms；
6) medicinal extract is sufficiently stirred with 27 tons 95 DEG C or more of hot water and uses ultrasonic oscillation 3 hours, high under (85 DEG C) while hot
25.7 tons of supernatant is collected by centrifugation in speed；
7) supernatant enters freezer, and refrigerated overnight, high speed centrifugation collects precipitation within second day, and vacuum drying obtains yellow greenish powder
638 kilograms of primary extract；
8) 638 kilograms of yellow green primary extracts are isolated and purified with the myricetin and dihydromyricetin of 98% or more high-purity of extraction
Sterling；
8.1) the first isolation and purification method；
8.1.1 it is (every to analyze pure ethyl acetates point 3 times with 2700 kilograms for) 319 kilograms of yellow green primary extract obtained by step 7)
Secondary 900 kilograms) amount is added in yellow greenish powder, and it constantly mixes slowly and with ultrasonic vibration 3 hours, ethyl acetate layer became yellowish green
Color, 3 times of ethyl acetate layers of last time are basically unchanged color；
8.1.2 (1) 218 kilogram of white depositions) are respectively obtained and ethyl acetate supernatant combined wt 2770 is public three times
Jin；
8.1.3 dissolving, high speed centrifugation, supernatant acidification refrigeration) are easy to 1000 kilogram of 5% sodium bicarbonate of precipitation (1)
Overnight white crystals precipitation dry at least 128.6 kilogram 90% purity dihydromyricetin, white, needle-shaped crystals are public with 600
95 DEG C of jin or more hot water is sufficiently stirred dissolving, and the precipitation that is centrifuged at a high speed collects supernatant, supernatant refrigerated overnight high speed centrifugation
Collect white, needle-shaped crystals, after pure water 3 times yellow needles drying can obtain 98% or more white needles dihydro poplar
123.9 kilograms of syphilis crystal.
8.1.4) 2770 kilograms of ethyl acetate supernatant；Supernatant recycles ethyl acetate, and vacuum drying obtains yellow powder 8.8
Kilogram, dissolving high speed centrifugation removing precipitation is easy using 100 kilogram of 5% sodium bicarbonate to yellow powder and collects supernatant 112.3
Kilogram, supernatant acidification is chilled overnight precipitation and separation and vacuum drying can obtain the needle-shaped myricetin of 98% or more yellow green and crystallize 8.4
Kilogram；
8.2) second of isolation and purification method；
8.2.1) 319 kilograms of yellow greenish powder primary extract obtained by step 7), it is molten to be added 3000 kilogram weights of 5% boric acid
Liquid is sufficiently stirred simultaneously ultrasonic vibration 3 hours, is then allowed to stand overnight precipitation；
8.2.2) rear high speed centrifugation removing precipitates to obtain 3350 kilograms of supernatant overnight；
8.2.3) 5% calcium hydroxide solution is added in supernatant, is slowly stirred, and constantly generates white flock precipitate, adjusts steady
Fixed slowly to be stirred more than half an hour until PH7.5, high speed centrifugation removes precipitation, collects 3030 kilograms of supernatant；
8.2.4 the supernatant) being collected into adjusts PH to 4.0, refrigerated overnight with hydrochloric acid；
8.2.5) rear high speed centrifugation precipitation, collection obtain the precipitation of the mixture of only myricetin and dihydromyricetin overnight
(2) 317 kilograms；
8.2.6) precipitation (2) detach myricetin and dihydromyricetin with 2700 kilograms of analysis pure ethyl acetate, and 900
Kilogram primary analysis pure ethyl acetate point is sufficiently stirred for 3 times and ultrasonic oscillation 3 hours or more, and high speed centrifugation precipitation collects supernatant
Liquid, precipitation with 900 kilograms of ethyl acetate ultrasounds and stirs be centrifuged at a high speed again precipitation and supernatant again, for the third time with 900
Kilogram ethyl acetate is stripped and centrifuges to precipitation；
8.2.7 the dihydromyricetin of white needle-like crystals is obtained after the crystallization drying of (3) 133.3 kg whites) must be precipitated
124.1 kilograms, 2720 kilograms of the supernatant three times of merging；
8.2.8) vacuum recycling ethyl acetate obtains 23.7 kilograms of body of paste, and 250 kilograms of 95 degree or more hot water are added in body of paste
Abundant dissolution filter, it is cold go to stay overnight high speed centrifugation collect yellow needles body and vacuum drying and obtain 8.4 kilograms of myricetin purity
98% or more.
Contrast experiment
Xiangxi Zhangjiajie vine tea analyzes general flavone content through us and reaches 36.5%, and wherein dihydromyricetin contains
Amount is 27.8%, and red bayberry cellulose content is 2.3%, and by searching for data, there is presently no a kind of commercial runs of maturation to extract simultaneously
Myricetin and dihydromyricetin, red bayberry cellulose content is low in industrial vine tea, generally extracts myricetin from red bayberry, in document also not
There are the extraction process of industrialization high-purity myricetin and dihydromyricetin, the present invention to have pioneering and initiative.
Experimental example 1
The content assaying method (Deng Danwen, University Of Nanchang's Food Science and technology National Key Laboratory) of myricetin
1, instrument
Aglient1260 high performance liquid chromatographs (quaternary pump, UV detector), 18 solid phases of Simon Aldrich-C extraction
Pillar is taken, JT2003 precise electronics balance (the Hebei road limited branch company of instrument highway instrument), the swing high speeds of DFY-500 are omnipotent
Pulverizer (Lin great Machinery Co., Ltd.s of Wenling city), Rotary Evaporators RE-52 (Shanghai Yarong Biochemical Instrument Plant), DHG-9053
Type electric heating constant-temperature blowing drying box (sends out scientific instrument Co., Ltd in Shanghai three), DKS-24 type electric-heated thermostatic water bath (Jiaxing Cities
Zhong Xin Medical Instruments Co., Ltd).
2, chromatographic condition
HPLC testing conditions chromatographic columns:18 liquid-phase chromatographic columns of Thermo-C (4.6mm × 250mm, 5 μm)；Mobile phase:
0.005mol/L potassium dihydrogen phosphates-methanol (volume ratio 30: 70)；Flow velocity is 0.5mL/min, 10 μ L of sample introduction, 25 DEG C of column temperature,
Detection wavelength 360nm.
3 Specification Curve of Increasing
It accurately weighs myricetin mark product 10mg to set in 100mL volumetric flasks, with methanol dissolving and constant volume, product are molten as a contrast
Liquid.With methanol dilution and constant volume, it is made into the myricetin mark product solution of 10,20,30,40,50 μ g/mL series concentrations, 10 μ L sample introductions,
It is analyzed into HPLC under 1.3.5.2 chromatographic conditions, using peak area as ordinate (Y), it is bent that standard is drawn with a concentration of abscissa (X)
Line.Gained regression equation is Y=54.155X-161.47, R=0.999 6, shows that sample linearly closes in 10~50 μ g/mL
System is good.
Experimental example 2
Dihydromyricetin content assaying method (Wang Jiasheng, Chinese patent drug in January, 2012 Guizhou University's Life Science College)
1, laboratory apparatus
Test apparatus and equipment mainly have KQ-500DE types ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.), point
Balance (ten a ten thousandths, plum Teller-support benefit Instrument Ltd.) is analysed, (Shanghai river letter science and technology has multifunctional crusher LX-02
Limit company)；Agilent1200LC (Anjelen Sci. ＆ Tech. Inc of the U.S.)；Dikma chromatographic columns (250mm × 4.60mm, 5 μm,
Chinese Di Ma companies).
2, chromatographic condition
10 μ L of chromatographic condition sample size；Ultraviolet detection wavelength 292nm；40 DEG C of column temperature；Mobile phase is 0.1% phosphoric acid-methanol-
Acetonihile gradient elution (0~10min, 74.4: 8: 17.6；10~15min, 36: 20: 44；15.1~18min, 74.4: 8:
17.6)；Volume flow 1mL/min.
3, the preparation of reference substance solution
Precision weighs dihydromyricetin 100.00mg, is placed in 100mL brown measuring bottles, is dissolved with methanol and is diluted to quarter
Degree, shake up, obtain mass concentration be 1.00mg/mL dihydromyricetin reference substance solution, respectively measure 1.0,2.0,4.0,
5.0,8.0,10.0mL, methanol constant volume to 10mL obtain control series product solution.
4, linear relationship
10 μ L of reference substance solution sample introduction, measure peak area, and each mass concentration sample introduction 3 times is averaged.With sample introduction quality
A concentration of abscissa (X), peak area are ordinate (Y), draw standard curve, and the regression equation for obtaining dihydromyricetin is Y=2
832.7X-101.24 (r2=0.996 0), the range of linearity are 0.5~10.0 μ g, show dihydromyricetin line within this range
Sexual intercourse is good.
5, sample measures
10 μ l of test solution sample introduction obtain sample chromatogram figure, pass through the amount of regression equation calculation dihydromyricetin.
Table：Testing result
| Sample | Myricetin | Dihydromyricetin |
| Embodiment 1 | Method 1,98.3% method 2,98.5% | Method 1,98.7%, method 2,99.1% |
| Embodiment 2 | Method 1,98.0% method 2,98.1% | Method 1,99.2%, method 2,98.9% |
| Embodiment 3 | Method 1,98.4% method 2,98.5% | Method 1,99.0%, method 2,99.1% |
Embodiment of above is merely to illustrate the present invention rather than limitation of the present invention.Although with reference to embodiment to this hair
It is bright to be described in detail, it will be understood by those of ordinary skill in the art that, to technical scheme of the present invention carry out it is various combination,
Modification or equivalent replacement, without departure from the spirit and scope of technical solution of the present invention, the right that should all cover in the present invention is wanted
It asks in range.
Claims (8)
1. a kind of method of separating-purifying myricetin and dihydromyricetin from vine tea, which is characterized in that this method includes as follows
Step：
Step 1：Preparation：Vine tea is weighed, it is selected, it crushes, crosses 35-45 mesh sieve；
Step 2：Extraction：The alcohol of the 75-85% of 18-22 times of vine tea weight is added, pH value is adjusted with 5% sodium bicarbonate solution
For 7.0-8.0, impregnate 6-10 hours, then reflux extracts 3-5 hours at a temperature of 35-45 DEG C；
Step 3：Secondary extraction：The alcohol of the 75-85% of 8-12 times of vine tea weight is added, flow back extraction at a temperature of 35-45 DEG C
3-5 hours；
Step 4：Filtering：Merging extracts liquid twice, and filtrate is obtained by filtration；
Step 5：Recovered alcohol：Recovered alcohol is compared for 15% hereinafter, being then concentrated in vacuo to alcohol concentration from filtrate
The medicinal extract of weight 1.0-1.5；
Step 6：Water is heavy：Concentrated extract is sufficiently stirred with 90-98 DEG C of hot water of 18-22 times of weight and uses ultrasonic oscillation
2.5-3.5 hours, high speed centrifugation at a temperature of 80-90 DEG C collected supernatant；
Step 7：Centrifugation：The supernatant refrigerated overnight that step 6 is obtained carries out high speed centrifugation in second day, collects precipitation, vacuum
Dry to obtain yellow greenish powder primary extract；
Step 8：It isolates and purifies：To the primary extract that step 7 obtains isolate and purify to obtain 98% or more high-purity myricetin and
Dihydromyricetin sterling, the purification procedures are as follows：
1) yellow greenish powder is added in the ethyl acetate of the yellow greenish powder primary extract that will be obtained in step 7, point 3 use, 9 times of weight
In primary extract, uniform stirring and with ultrasonic vibration 2.5-3.5 hours, ethyl acetate layer becomes yellow green, merges and obtains three times
Ethyl acetate supernatant and the white depositions obtained three times；
2) 8-12 times of 5% sodium bicarbonate solution of white depositions that step 1) obtains being dissolved, high speed centrifugation takes supernatant,
Supernatant is acidified, refrigerated overnight, obtains white crystals precipitation, drying obtains the dihydromyricetin of purity at least 90%；Again will
90 DEG C or more the hot water that white crystals precipitate 5 times of weight are sufficiently stirred dissolving, high speed centrifugation, and precipitation and separation collects supernatant,
By supernatant refrigerated overnight, high speed centrifugation, collection obtains white, needle-shaped crystals, with pure water 3 times, obtains white needles knot
Crystalline substance, drying obtain the white needles dihydromyricetin cellulose crystal of 98% or more purity；
3) ethyl acetate is recycled in the supernatant obtained from step 1), vacuum drying obtains yellow powder, and yellow powder is used
5% sodium bicarbonate solution dissolves, high speed centrifugation, removes precipitation and collects supernatant, supernatant is acidified, is chilled overnight, separation is heavy
The needle-shaped myricetin crystallization of 98% or more yellow green can be obtained by forming sediment and being dried in vacuo.
2. the method for separating-purifying myricetin and dihydromyricetin according to claim 1 from vine tea, which is characterized in that
In step 8, the purification procedures are as follows：
1) the yellow greenish powder primary extract that will be obtained in step 7 is added 5% boric acid solution of 10 times of primary extract weight, fully stirs
Simultaneously ultrasonic vibration 2.5-3.5 hours are mixed, overnight precipitation is then allowed to stand；High speed centrifugation removes precipitation, collects supernatant；
2) 5% calcium hydroxide solution is added in supernatant, is slowly stirred, constantly generates white flock precipitate, adjust stablize to
Until pH 7.5, then slowly more than half an hour, high speed centrifugation removes precipitation for agitation, collects supernatant；
3) by the supernatant being collected into salt acid for adjusting pH value to 4.0, refrigerated overnight, high speed centrifugation precipitation, collection only had
The precipitation of the mixture of myricetin and dihydromyricetin；
4) it is added three times the ethyl acetate of 9 times of weight in the precipitation that step 3) obtains, is sufficiently stirred and ultrasonic oscillation 3 is small
When more than, stand overnight；High speed centrifugation collects precipitation and supernatant respectively；
5) the white needles dihydromyricetin cellulose crystal for precipitating vacuum drying to get purity 98% for obtaining step 4)；
6) vacuum recycles ethyl acetate in the supernatant obtained from step 4), obtains body of paste, and 10 times of 95 DEG C or more hot water are added,
Fully dissolving is filtered, cooling, and overnight, high speed centrifugation, collection obtains the yellow needles red bayberry cellulose crystal of 98% or more purity.
3. the method for separating-purifying myricetin and dihydromyricetin according to claim 1 from vine tea, which is characterized in that
In step 1, the sieve is 40 mesh.
4. the method for separating-purifying myricetin and dihydromyricetin according to claim 1 from vine tea, which is characterized in that
In step 2 and step 3, the extraction temperature is 40 DEG C.
5. the method for separating-purifying myricetin and dihydromyricetin according to claim 1 from vine tea, which is characterized in that
In step 5, the proportion is 1.15-1.20.
6. the method for separating-purifying myricetin and dihydromyricetin according to claim 1 from vine tea, which is characterized in that
In step 6, the heavy temperature of the water is 95 DEG C.
7. the method for separating-purifying myricetin and dihydromyricetin according to claim 1 from vine tea, which is characterized in that
In the step 3) of step 8, vacuum degree is -0.08--0.09MPa.
8. according to the method for claim 1-7 any one of them separating-purifying myricetin and dihydromyricetin from vine tea,
It is characterized in that, this method comprises the following steps：
Step 1：Preparation：Vine tea is weighed, it is selected, it crushes, crosses 40 mesh sieve；
Step 2：Extraction：80% alcohol of 20 times of vine tea weight is added, adjusting pH value with 5% sodium bicarbonate solution is
7.5, it impregnates 8 hours, then reflux extracts 4 hours at a temperature of 40 DEG C；
Step 3：Secondary extraction：80% alcohol of 10 times of vine tea weight is added, reflux extraction 4 hours at a temperature of 40 DEG C；
Step 4：Filtering：Merging extracts liquid twice, and filtrate is obtained by filtration；
Step 5：Recovered alcohol：Recovered alcohol is compared for 15% hereinafter, being then concentrated in vacuo to alcohol content from filtrate
Weigh 1.2 medicinal extract；
Step 6：Water is heavy：95 DEG C of hot water of 20 times of weight of concentrated extract is sufficiently stirred and uses ultrasonic oscillation 3 hours,
High speed centrifugation at a temperature of 85 DEG C collects supernatant；
Step 7：Centrifugation：The supernatant refrigerated overnight that step 6 is obtained carries out high speed centrifugation in second day, collects precipitation, vacuum
Dry to obtain yellow greenish powder primary extract；
Step 8：It isolates and purifies：To the primary extract that step 7 obtains isolate and purify to obtain 98% or more high-purity myricetin and
Dihydromyricetin sterling.
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Denomination of invention: Method for separating and purifying myricetin and dihydromyricetin from ampelopsis grossedentata Effective date of registration: 20191125 Granted publication date: 20180928 Pledgee: Bank of Changsha Co., Ltd. Kaifu sub branch Pledgor: HUNAN NUTRAMAX INC. Registration number: Y2019430000047 |
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Date of cancellation: 20201217 Granted publication date: 20180928 Pledgee: Bank of Changsha Co.,Ltd. Kaifu sub branch Pledgor: HUNAN NUTRAMAX Inc. Registration number: Y2019430000047 |
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Denomination of invention: A method for separation and purification of myricetin and Dihydromyricetin from Ampelopsis grossedentata Effective date of registration: 20201218 Granted publication date: 20180928 Pledgee: Bank of Changsha Co.,Ltd. Kaifu sub branch Pledgor: HUNAN NUTRAMAX Inc. Registration number: Y2020980009579 |
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Date of cancellation: 20220615 Granted publication date: 20180928 Pledgee: Bank of Changsha Co.,Ltd. Kaifu sub branch Pledgor: HUNAN NUTRAMAX Inc. Registration number: Y2020980009579 |