CN100484948C - Method for separating derivative of vitexin - Google Patents
Method for separating derivative of vitexin Download PDFInfo
- Publication number
- CN100484948C CN100484948C CNB2007100112416A CN200710011241A CN100484948C CN 100484948 C CN100484948 C CN 100484948C CN B2007100112416 A CNB2007100112416 A CN B2007100112416A CN 200710011241 A CN200710011241 A CN 200710011241A CN 100484948 C CN100484948 C CN 100484948C
- Authority
- CN
- China
- Prior art keywords
- vitexin
- glucoside
- alcohol
- rhamnoside
- silica gel
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Saccharide Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
This invention discloses a method for separating vitexin derivatives, vitexin-4'-O-glucoside and vitexin-2''-O-glucoside. The method comprises: (1) pulverizing hawthorn leaves into coarse powder, extracting by refluxing in alcohol aqueous solution, filtering, incorporating the filtrates, vacuum-concentrating to no alcohol odor, removing impurities with macroporous adsorption resin AB-8 or D101, eluting with 30-80% ethanol, vacuum-concentrating to no alcohol odor, decolorizing with petroleum ether, extracting with n-butanol, vacuum-recovering n-butanol, separating the extract by silica gel column chromatography, eluting with chloroform-methanol at a ratio of (4-3):1, vacuum-concentrating the eluate, separating by silica gel column chromatography,, eluting with ethyl acetate-butanone-methanoic acid-water or ethyl acetate-acetone-methanoic acid-water, and standing at 0-15 deg.C to obtain vitexin-4'-O-glucoside and vitexin-2''-O-glucoside. The method has such advantages as high yield, high purity, simple process and low toxicity, and is environmentally friendly.
Description
Technical field
The present invention relates to separate a kind of derivative of vitexin by the ordinary silicon plastic column chromatography: Vitexin-4 '-method of O-glucoside and vitexin-2 ''-O-rhamnoside, what The present invention be more particularly directed to is to adopt silica gel column chromatography (used filler is a tlc silica gel), with ethyl acetate-butanone (or acetone)-formic acid-water is eluent, from Fruit of Pashi Pear leaf or Folium Crataegi extract, directly separate Vitexin-4 '-O-glucoside and Vitexin-" method of O-rhamnoside.
Background technology
Howthorn Leaf is the dry leave of rosaceous plant Fruit of Pashi Pear (Crataegus pinnatifida Bge.var major N.E.Br.), hawthorn (C.pinnatifida Bge.), records in the Pharmacopoeia of the People's Republic of China 2005 editions, 22 pages.Have promoting blood circulation and removing blood stasisly, regulate the flow of vital energy and promote blood circulation, be used for qi depression to blood stasis, chest distress, palpitaition is seen and is forgotten, dizzy tinnitus.Result of study shows derivative of vitexin: Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside are to be proved the treatment cardiovascular disorder, and the synthetic and antioxygenation of the very strong inhibition mankind's breast cancer cell DNA is arranged.Silica gel chromatography is fit to the separating flavone constituents, therefore the present invention is with the silica gel column chromatography technology and in conjunction with the thin-layer chromatography filler, successfully be used to separate Vitexin-4 '-O-glucoside and Vitexin-" the O-rhamnoside, two kinds of compounds can be used as the raw material of treatment relative disease medicine.
At present Vitexin-4 '-separation method of O-glucoside and vitexin-2 ''-O-rhamnoside mainly adopts alcohol extracting, the solvent extraction method of binding silica gel column chromatography simultaneously.The bibliographical information characteristics are cost height, and the cycle is long, and yield is low.[Fruit of Pashi Pear phyllody composition Study Chinese medicinal materials such as Liu Ronghua, 2006,29 (11); Howthorn Leaf The Chemical Constituents CHINA JOURNAL OF CHINESE MATERIA MEDICAs such as fourth apricot bud, 1990; 15 (5)]
Summary of the invention
The invention provides easy, environmental protection, fast, the purity height, a kind of derivative of vitexin that productive rate is high: Vitexin-4 '-separation method of O-glucoside and vitexin-2 ''-O-rhamnoside.
The object of the present invention is achieved like this:
A kind of separation derivative of vitexin: Vitexin-4 '-method of O-glucoside and vitexin-2 ''-O-rhamnoside, be to get Fruit of Pashi Pear leaf or Howthorn Leaf medicinal material, be ground into meal, use the aqueous alcohol refluxing extraction, extracting liquid filtering, merging filtrate, be evaporated to medicinal extract and do not have the alcohol flavor, through the removal of impurities of AB-8 (D101) macroporous adsorbent resin, end to no longer detecting flavones with the 30-80% ethanol elution, behind the sherwood oil depigmentation, colourless to elutriant with n-butanol extraction again, the reclaim under reduced pressure propyl carbinol is to doing, and the dry thing of gained separates through silica gel column chromatography (wherein silica gel is with the 200-260 order), use the chloroform-methanol wash-out, with chloroform, all flow points of methyl alcohol (4-3:1) position (1-2 column volume) elutriant is evaporated to dried, and dry thing is eluted to two visible main colour bands with ethyl acetate-butanone (or acetone)-formic acid-water (4-3:3:1:1) and flows out again through the tlc silica gel column chromatography, directly separate out crystallization under 0-15 ℃ of temperature, purity is 90-99%.
Fruit of Pashi Pear leaf or Howthorn Leaf meal according to actual production, can adopt extraction using alcohol to be advisable, consumption be medicinal material 8-16 doubly, can adopt modes such as cold soaking, ultrasonic and refluxing extraction, wherein the mode of reflux is a preferred version.
In the refluxing extraction operation of aqueous alcohol, the concentration of aqueous alcohol is 30-80%, and consumption is 8-16 a times of medicinal material, refluxing extraction 2-3 time, and each 1-3 hour, alcohol can be selected methyl alcohol or ethanol for use, according to actual production and toxicity, selects for use ethanol the most suitable.
Alcohol extract filters, and being evaporated to does not have the alcohol flavor, through the removal of impurities of AB-8 (D101) macroporous adsorbent resin, with the 30-80% ethanol elution, behind the sherwood oil depigmentation, use n-butanol extraction again, the reclaim under reduced pressure propyl carbinol extract obtainedly separates through silica gel column chromatography (wherein silica gel is with the 200-260 order).
Silicagel column chloroform-methanol wash-out, all flow points of chloroform-methanol (4-3:1) position (1-2 column volume) eluate concentrating under reduced pressure, dry thing separates through silica gel column chromatography again, wherein filler is a tlc silica gel, flow out with the main visible colour band of ethyl acetate-butanone (or acetone)-formic acid-water elution to two, under 0-15 ℃ of temperature, directly separate out Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside crystallization.Ketone can be selected butanone or acetone for use, adopts ethyl acetate-butanone (or acetone)-formic acid-water (4-3:3:1:1).
Present method prepared Vitexin-4 '-extract yield of O-glucoside and vitexin-2 ''-O-rhamnoside is 0.015-0.05%, Vitexin-4 '-content of O-glucoside and vitexin-2 ''-O-rhamnoside is 90-99% (adopt high performance liquid chromatography, normalization method is measured).This extract can separately or be united as bulk drug and is applied in the healing potion of various relative diseases.
What the present invention was different with existing document is: adopt single silica gel column chromatography to divide to be the derivative of vitexin in separable Fruit of Pashi Pear leaf or the Howthorn Leaf: Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside, avoided operational difficulty, shortcoming that cost is high.Use this law isolating Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside yield height, purity height, little, the environmental protection of toxicity.Present method is easy, quick, practical, is fit to suitability for industrialized production.
Embodiment
Embodiment one
The Fruit of Pashi Pear leaf is ground into meal, with 10 times of amount 70% alcohol reflux 3 times, is respectively 2,1,1 hours at every turn, filter, and merging filtrate, being evaporated to does not have the alcohol flavor.Gained medicinal extract no longer detects through AB-8 macroporous adsorbent resin removal of impurities to sugar, end to no longer detecting flavones with 50% ethanol elution again, alcohol eluen is evaporated to does not have the alcohol flavor, behind the sherwood oil depigmentation, n-butanol extraction is colourless to elutriant, the reclaim under reduced pressure propyl carbinol is to doing, the dry thing of gained separates through silica gel column chromatography (wherein silica gel is with the 200-260 order), use the chloroform-methanol wash-out, with chloroform, all flow points of methyl alcohol (4-3:1) position (1-2 column volume) eluate concentrating under reduced pressure, dry thing is through the tlc silica gel post, being eluted to two main visible colour bands with ethyl acetate-butanone-formic acid-water (4:3:1:1) flows out, under 0-15 ℃ of temperature, directly separate out Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside crystallization, purity is (through high performance liquid chromatography, normalization method is measured) all greater than 90%.
Embodiment two
Howthorn Leaf is ground into meal, with 10 times of amount 50% alcohol reflux 2 times, and each 2 hours, filter, merging filtrate, being evaporated to does not have the alcohol flavor.Gained medicinal extract no longer detects through D101 macroporous adsorbent resin removal of impurities to sugar, end to no longer detecting flavones with 60% ethanol elution again, alcohol eluen is evaporated to does not have the alcohol flavor, behind the sherwood oil depigmentation, n-butanol extraction is colourless to elutriant, the reclaim under reduced pressure propyl carbinol, dry, the dry thing of gained separates through silica gel column chromatography (wherein silica gel is with the 200-260 order), use the chloroform-methanol wash-out, with chloroform, all flow points of methyl alcohol (4-3:1) position (1-2 column volume) eluate concentrating under reduced pressure, dry thing is eluted to two main visible colour bands with ethyl acetate-acetone-formic acid-water (3:3:1:1) and flows out through the tlc silica gel post, under 0-15 ℃ of temperature, directly separate out Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside crystallization, purity is 99% (through high performance liquid chromatography, normalization method is measured).
Embodiment three
The Fruit of Pashi Pear leaf is ground into meal, with 16 times of amount 60% alcohol reflux 2 times, and each 2 hours, filter, merging filtrate, being evaporated to does not have the alcohol flavor.Gained medicinal extract no longer detects through AB-8 macroporous adsorbent resin removal of impurities to sugar, end to no longer detecting flavones with 70% ethanol elution again, alcohol eluen is evaporated to does not have the alcohol flavor, behind the sherwood oil depigmentation, n-butanol extraction is colourless to elutriant, the reclaim under reduced pressure propyl carbinol, dry, the dry thing of gained separates through silica gel column chromatography (wherein silica gel is with the 200-260 order), use the chloroform-methanol wash-out, with chloroform, all flow points of methyl alcohol (4-3:1) position (1-2 column volume) eluate concentrating under reduced pressure, dry thing is eluted to two main visible colour bands with ethyl acetate-butanone-formic acid-water (4:3:1:1) and flows out through the tlc silica gel post, under 0-15 ℃ of temperature, directly separate out Vitexin-4 '-O-glucoside and vitexin-2 ''-O-rhamnoside crystallization, purity is all greater than 95% (through high performance liquid chromatography, normalization method is measured).
Claims (4)
1, a kind of separation derivative of vitexin: the method for Vitexin-4 ＇-O-glucoside and vitexin-2 ''-O-rhamnoside, it is characterized in that getting Fruit of Pashi Pear leaf and Howthorn Leaf medicinal material, be ground into meal, with aqueous alcohol concentration is 30-80%, the water alcohol reflux extracts 2-3 time, each 1-3 hour, consumption is that the 8-16 of medicinal material doubly measures, extracting liquid filtering, merging filtrate, be evaporated to and do not have the alcohol flavor, through AB-8 or the removal of impurities of D101 macroporous adsorbent resin, end to no longer detecting flavones with the 30-80% ethanol elution, alcohol eluen is evaporated to does not have the alcohol flavor, behind the sherwood oil depigmentation, colourless with n-butanol extraction again to elutriant, the reclaim under reduced pressure propyl carbinol is to doing, the dry thing of gained separates through 200-260 order silica gel column chromatography, uses the chloroform-methanol wash-out, chloroform, the methyl alcohol ratio of components is 4-3:1, consumption is 1-2 a times of column volume, elutriant is evaporated to dried, the dry thing of gained is through silica gel column chromatography, and filler is a tlc silica gel, flow out with ethyl acetate-butanone-formic acid-water elution or ethyl acetate-acetone-formic acid-water elution to two main visible colour band again, under 0-15 ℃ of temperature, directly separate out Vitexin-4 ＇-O-glucoside and vitexin-2 ''-O-rhamnoside crystallization.
2, require 1 described a kind of separation derivative of vitexin according to letter of authorization: the method for Vitexin-4 ＇-O-glucoside and vitexin-2 ''-O-rhamnoside is characterized in that described aqueous alcohol is an ethanolic soln.
3, require 1 described a kind of separation derivative of vitexin according to letter of authorization: the method for Vitexin-4 ＇-O-glucoside and vitexin-2 ''-O-rhamnoside, the ratio of components that it is characterized in that ethyl acetate-butanone-formic acid-water or ethyl acetate-acetone-formic acid-water is 4-3:3:1:1.
4, a kind of separation derivative of vitexin according to claim 1: the method for Vitexin-4 ＇-O-glucoside and vitexin-2 ''-O-rhamnoside, it is characterized in that gained Vitexin-4 ＇-O-glucoside and vitexin-2 ''-O-rhamnoside purity through high performance liquid chromatography, normalization method is measured and is 90-99%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100112416A CN100484948C (en) | 2007-05-10 | 2007-05-10 | Method for separating derivative of vitexin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100112416A CN100484948C (en) | 2007-05-10 | 2007-05-10 | Method for separating derivative of vitexin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101050226A CN101050226A (en) | 2007-10-10 |
| CN100484948C true CN100484948C (en) | 2009-05-06 |
Family
ID=38781848
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2007100112416A Expired - Fee Related CN100484948C (en) | 2007-05-10 | 2007-05-10 | Method for separating derivative of vitexin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100484948C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102234299A (en) * | 2010-04-28 | 2011-11-09 | 辽宁中医药大学 | Separation method of hyperin |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101849918A (en) * | 2010-05-27 | 2010-10-06 | 辽宁中医药大学 | Vitexin-2''-O-rhamnoside pill and preparation method |
| CN101879436B (en) * | 2010-07-15 | 2011-11-30 | 南京工业大学 | Decolorizing column and method for performing normal temperature decolorization on DHA grease by employing same |
| CN106265706A (en) * | 2015-05-20 | 2017-01-04 | 辽宁中医药大学 | A kind of male ice drop pill treating acute myocardial ischemia and preparation method thereof |
| CN104974202B (en) * | 2015-06-26 | 2017-11-28 | 广西中医药大学 | A kind of method of the extraction separation Vitexin xyloside from natural products |
| CN105254691B (en) * | 2015-10-27 | 2018-07-06 | 中国科学院华南植物园 | A kind of method for preparing 2 "-O- β-D-xylosylvitexin |
| CN108689849B (en) * | 2018-07-13 | 2021-08-31 | 鲁东大学 | Method for simultaneously separating flavonoid and chlorogenic acid compounds from hawthorn leaf crude extract |
| IL275222A (en) * | 2020-06-08 | 2022-01-01 | The Open Univ | The chinese herb shan-zha for the treatment of depression and anxiety disorders; efficiency, safety and biological correlates |
-
2007
- 2007-05-10 CN CNB2007100112416A patent/CN100484948C/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 山楂叶化学成分的研究. 丁杏苞等.中国中药杂质，第15卷第5期. 1990 |
| 山楂叶化学成分的研究. 丁杏苞等.中国中药杂质，第15卷第5期. 1990 * |
| 山楂叶植物学及化学成分研究近况. 英锡相等.辽宁中医学院学报，第3卷第2期. 2001 |
| 山楂叶植物学及化学成分研究近况. 英锡相等.辽宁中医学院学报，第3卷第2期. 2001 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102234299A (en) * | 2010-04-28 | 2011-11-09 | 辽宁中医药大学 | Separation method of hyperin |
| CN102234299B (en) * | 2010-04-28 | 2014-10-08 | 辽宁中医药大学 | Separation method of hyperin |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101050226A (en) | 2007-10-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100484948C (en) | Method for separating derivative of vitexin | |
| CN102451235B (en) | Preparation method of olive leaf extract | |
| CN101220062A (en) | Method for preparing stevioside and rebaudiodside A simultaneously | |
| CN102258588B (en) | Preparation method of peony general glycoside | |
| CN101985421A (en) | Method for simultaneously preparing chlorogenic acid and luteoloside from honeysuckle flower | |
| CN101690739A (en) | Preparation method of dogbane leaf extractive | |
| CN101259159A (en) | Preparation of alhagi sparsifolia total flavones and uses thereof | |
| CN107998212A (en) | A kind of preparation method of glutinous rehmannia cyclic olefine ether terpinyl side like extractive | |
| CN102746362A (en) | Method for extracting refined astragaloside from astragaliradix | |
| CN103483402A (en) | Method for purifying and preparing stevioside and rebaudioside-A | |
| CN103819445A (en) | Method for preparing two neo-pentenyl flavonoid compounds with hypolipidemic activity in fructus podophylli | |
| CN102552371B (en) | Gynostemma pentaphylla extractive and application thereof to preparing medicine for treating tumor | |
| CN101348474A (en) | Method for preparing salvianolic acid B and tanshinol from Salvia miltiorrhiza stem | |
| CN102180938A (en) | Method for preparing capilliposide | |
| CN109879919B (en) | Method for separating and preparing three flavonoid glycosides from spina date seeds | |
| CN102228488A (en) | Preparation of Lysimachia capillipes Hemsl total saponin | |
| CN100586449C (en) | Method for preparing white peony root total glycosides extract | |
| CN106045960B (en) | A kind of method that Diphenylheptane class substance is extracted from galangal | |
| CN102503996B (en) | Method for extracting active constituent from Swertia mussotii plant | |
| CN102464693B (en) | Ginsenoside Re extraction and separation method | |
| CN102002087A (en) | Method for preparing bryonia alcohol acid | |
| CN102234299B (en) | Separation method of hyperin | |
| CN101696381B (en) | Novel process for preparing highland barley flavone extract and application thereof in health wine | |
| CN103951717B (en) | The method of benzoylpaeoniflorin and benzoyl lactone glucoside of Radix Paeoniae is prepared in a kind of extraction | |
| CN109776515A (en) | The method of mangiferin is extracted from myrica rubra leaf |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Dalian Tianyu Olson Pharmaceutical Co. Ltd. Assignor: Liaoning University of TCM Contract fulfillment period: 2009.10.26 to 2014.10.25 contract change Contract record no.: 2009210000269 Denomination of invention: Method for separating derivative of vitexin Granted publication date: 20090506 License type: Exclusive license Record date: 20091110 |
|
| LIC | Patent licence contract for exploitation submitted for record |
Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2009.10.26 TO 2014.10.25; CHANGE OF CONTRACT Name of requester: DALIAN TIANYU AOSEN PHARMACEUTICAL CO., LTD. Effective date: 20091110 |
|
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090506 Termination date: 20140510 |