CN106336440A - Method for extracting and separating out oleanolic acid from olive leaves - Google Patents
Method for extracting and separating out oleanolic acid from olive leaves Download PDFInfo
- Publication number
- CN106336440A CN106336440A CN201610728088.8A CN201610728088A CN106336440A CN 106336440 A CN106336440 A CN 106336440A CN 201610728088 A CN201610728088 A CN 201610728088A CN 106336440 A CN106336440 A CN 106336440A
- Authority
- CN
- China
- Prior art keywords
- oleanolic acid
- solution
- sodium bicarbonate
- concentration
- growing leaves
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H13/00—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids
- C07H13/02—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids
- C07H13/08—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids having the esterifying carboxyl radicals directly attached to carbocyclic rings
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Compounds Of Unknown Constitution (AREA)
Abstract
The invention discloses a method for extracting and separating out oleanolic acid from olive leaves. The method includes the following steps of firstly, smashing the olive leaves to be extracted through an ethyl alcohol solution with volume concentration of 70-90%, filtering an extraction solution, and decompressing filtrate to recover ethyl alcohol to obtain crude extracts; secondly, adding the crude extracts into an ethyl alcohol solution with volume concentration of 50-65% to be dissolved again to obtain a mixed solution; thirdly, applying the mixed solution to an AB-8 type macroporous adsorption resin column, conducting eluting through water till being colorless, then conducting eluting through a sodium bicarbonate solution at temperature of 40-50 DEG C and concentration of 0.1-0.15 mol/L, conducting eluting through a sodium bicarbonate solution at temperature of 68-72 DEG C and concentration of 1-1.5 mol/L, conducting eluent obtained in three times, adjusting pH to be 3-5, conducting filtering, and taking filter residues to be dried to obtain oleanolic acid. Extracted oleanolic acid is high in purity and high in yield.
Description
Technical field
The invention belongs to technical field of biological extraction is and in particular to a kind of extraction from olive growing leaves separates oleanolic acid
Method.
Background technology
Olive (olea europare l.) is world-renowned woody oil tree species, has the cultivation of more than 4000 year to go through
History.The decoction of olive growing leaves or preserved material have certain curative effect in mediterranean country frequently as the medication among the people of diabetes.Olive
Leaf extract is used as dietary supplements to strengthen immunologic function in America and Europe.Modern data shows, contains in olive growing leaves
The active ingredients such as flavonoids, triterpenes, carbohydrate, row iridoids.Triterpene compound is mainly by 6 isoprene structures
Unit is constituted, and major part is the terpenoid containing 30 carbon atoms.In olive growing leaves, triterpenes components mainly include olive
Acid, crataegolic acid and ursolic acid, and these three structures of matter are similar, generally carried out separating using Column chromatography techniques, but repeatedly
Silica gel column chromatography separates and recrystallization leads to the sample size finally giving little.Again because Column chromatography techniques generally existing processes week
Phase is long, and continuous operation property is not strong, is difficult to be used for the industrial production process of bioactivator as separating medium.
Content of the invention
Present invention solves the technical problem that being to provide a kind of method extracting oleanolic acid from olive growing leaves, the method letter
Single, cycle is short, the oleanolic acid purity obtaining is high, and yield is big.
The technical scheme that the present invention provides is the method extracting oleanolic acid from olive growing leaves, comprises the following steps:
1), after olive growing leaves being pulverized, extracted with the ethanol solution that volumetric concentration is 70～90%, extract is filtered, takes
Decompression filtrate recycling ethanol, obtains crude extract；
2) crude extract addition volumetric concentration is dissolved again for 50～65% ethanol solutions, obtain mixed solution；
3) by ab-8 large pore resin absorption column on mixed solution, first wash with water to colourless, then adopt 40～50 DEG C,
The sodium bicarbonate solution of 0.1～0.15mol/l is eluted, then is entered with the sodium bicarbonate solution of 68～72 DEG C, 1～1.5mol/l
Row wash-out, collects third time eluent, adjusts ph to 3～5, filters, takes filter residue and drying, obtain final product.
In step 1) in, olive growing leaves are extracted using the ethanol solution that volumetric concentration is 70～90%, can abundant extract oil
Oleanolic acid in olive leaf, crataegolic acid and ursolic acid.Olive growing leaves are 1g:10～20ml with the solid-liquid ratio of ethanol solution.
Described be extracted as heating and refluxing extraction, extraction time be 1～3 time, each extraction time be 1～3h.
Step 2) in, the ethanol solution dissolving crude extract being 50～65% using volumetric concentration, pole in removable crude extract
The less impurity of property, plays the effect of preliminary purification.Ethanol solution to crude extract concentration is added to be 0.06～0.10g/ml, now
The adsorption effect of upper prop is optimal.
Step 3) in, the macroreticular resin of ab-8 model is low pole, and average pore size is 13～14nm, and specific surface area is 480
～520m2/g.Because oleanolic acid, crataegolic acid, ursolic acid are low pole material, and there is certain hydrophobicity, generate hydrogen
The ability of key is weaker, thus is easily adsorbed by nonpolar and low pole resin.Secondly, during macroporous absorbent resin adsorbent, quilt
Absorbed component is to be diffused into resin internal surface of hole by the aperture of resin and be adsorbed, when resin aperture is less than effective one-tenth of absorption
During fractionated molecule, active ingredient molecule can not enter inside resin, can only adsorb in resin outer surface, when resin aperture is more than effectively
During component molecules, active ingredient can be adsorbed and also be can be adsorbed on inner surface in resin outer surface, to crataegolic acid, oleanolic acid and
For ursolic acid, its molecular weight is respectively 472,456 and 456, therefore adopts the adsorption effect of ab-8 macroreticular resin preferable.Resin
The blade diameter length ratio of post is advisable for 1:3～6, optimal with 1:4.
Oleanolic acid, crataegolic acid and ursolic acid structure are very much like, and wherein ursolic acid and oleanolic acid are even more isomerism
Body, common column chromatography is difficult to be isolated and comes, but the present invention adopts 40～50 DEG C, the bicarbonate of 0.1～0.15mol/l
Sodium solution can be larger with polarity in resin hawthorn acid reaction generate soluble sodium salt, and then elute, and polarity be less than normal
Oleanolic acid and the carboxyl of ursolic acid and the polarizability of hydrone poor it is impossible to ionize out h+ it is impossible to enter with sodium acid carbonate
Row reaction generates sodium salt, then stay in resin column.The elution speed of sodium bicarbonate solution be 1～1.5bv/h, its consumption be 1～
During 1.5bv, wash-out effect is preferable.
Sodium bicarbonate solution using 68～72 DEG C, 1～1.5mol/l is eluted, and now can elute oleanolic acid,
And ursolic acid is not eluted, because oleanolic acid and ursolic acid structure are isomer, therefore, using elution speed faster
2～2.5bv/h, less amount of eluent is 0.5～1bv, at utmost can elute oleanolic acid, and ursolic acid is not eluted.
The method of the present invention is simple, cycle is short, and the oleanolic acid purity of extraction is high, and yield is big.
Specific embodiment
The present invention is further elaborated for specific examples below, but not as a limitation of the invention.
Embodiment 1
1) olive growing leaves are crushed to 100 mesh, the ethanol solution being 70% with volumetric concentration is by solid-liquid ratio for 1g:10ml's
Ratio mixes, and extract is filtered, takes decompression filtrate recycling ethanol, obtains crude extract by heating and refluxing extraction 3h；
2) crude extract being added volumetric concentration is 0.06g/ml for 50% ethanol solution to crude extract concentration, stirs evenly, obtains
Mixed solution；
3) by ab-8 large pore resin absorption column on mixed solution, resin column blade diameter length ratio is 1:3, first washes with water to colourless,
Be subsequently adding 40 DEG C, the sodium bicarbonate solution of 0.1mol/l eluted, the elution speed of sodium bicarbonate solution is 1bv/h, its
Consumption is 1bv, then is eluted with the sodium bicarbonate solution of 68 DEG C, 1mol/l, and elution speed is 2bv/h, and its consumption is
0.5bv, collects third time eluent (i.e. the sodium acid carbonate eluent of 1mol/l), adjusts ph to 3, filters, takes filter residue and drying, that is,
?.
It is measured according to high performance liquid chromatography, its purity is 99.1%, yield is 93.3%.
Embodiment 2
1) olive growing leaves are crushed to 180 mesh, the ethanol solution being 90% with volumetric concentration is by solid-liquid ratio for 1g:20ml's
Ratio mixes, heating and refluxing extraction 3 times, and each extraction time is 3h, extract is filtered, takes decompression filtrate recycling ethanol, obtain
To crude extract；
2) crude extract being added volumetric concentration is 0.10g/ml for 65% ethanol solution to crude extract concentration, stirs evenly, obtains
Mixed solution；
3) by ab-8 large pore resin absorption column on mixed solution, resin column blade diameter length ratio is 1:6, first washes with water to colourless,
Be subsequently adding 50 DEG C, the sodium bicarbonate solution of 0.15mol/l eluted, the elution speed of sodium bicarbonate solution is 1.5bv/h,
Its consumption is 1.5bv, then is eluted with the sodium bicarbonate solution of 72 DEG C, 1.5mol/l, and elution speed is 2.5bv/h, its use
Measure as 1bv, collect third time eluent (i.e. the sodium acid carbonate eluent of 1.5mol/l), adjust ph to 5, filter, take filter residue to do
Dry, obtain final product.
It is measured according to high performance liquid chromatography, its purity is 99.0%, yield is 93.0%.
Embodiment 3
1) olive growing leaves are crushed to 150 mesh, the ethanol solution being 90% with volumetric concentration is by solid-liquid ratio for 1g:20ml's
Ratio mixes, heating and refluxing extraction 2 times, and each extraction time is 2h, extract is filtered, takes decompression filtrate recycling ethanol, obtain
To crude extract；
2) crude extract being added volumetric concentration is 0.08g/ml for 55% ethanol solution to crude extract concentration, stirs evenly, obtains
Mixed solution；
3) by ab-8 large pore resin absorption column on mixed solution, resin column blade diameter length ratio is 1:4, first washes with water to colourless,
Be subsequently adding 45 DEG C, the sodium bicarbonate solution of 0.12mol/l eluted, the elution speed of sodium bicarbonate solution is 1.2bv/h,
Its consumption is 1.2bv, then is eluted with the sodium bicarbonate solution of 70 DEG C, 1.35mol/l, and elution speed is 2.5bv/h, its use
Measure as 0.5bv, collect third time eluent (the sodium acid carbonate eluent of 1.35mol/l), adjust ph to 4, filter, take filter residue to do
Dry, obtain final product.
It is measured according to high performance liquid chromatography, its purity is 99.5%, yield is 94.5%.
Embodiment 4
1) olive growing leaves are crushed to 200 mesh, the ethanol solution being 70% with volumetric concentration is by solid-liquid ratio for 1g:20ml's
Ratio mixes, heating and refluxing extraction 3 times, and each extraction time is 1h, extract is filtered, takes decompression filtrate recycling ethanol, obtain
To crude extract；
2) crude extract being added volumetric concentration is 0.06g/ml for 65% ethanol solution to crude extract concentration, stirs evenly, obtains
Mixed solution；
3) by ab-8 large pore resin absorption column on mixed solution, resin column blade diameter length ratio is 1:3, first washes with water to colourless,
Be subsequently adding 50 DEG C, the sodium bicarbonate solution of 0.1mol/l eluted, the elution speed of sodium bicarbonate solution is 1.5bv/h,
Its consumption is 1bv, then is eluted with the sodium bicarbonate solution of 68 DEG C, 1.5mol/l, and elution speed is 2bv/h, and its consumption is
1bv, collects third time eluent (i.e. the sodium acid carbonate eluent of 1.5mol/l), adjusts ph to 3～5, filters, and takes filter residue to do
Dry, obtain final product.
It is measured according to high performance liquid chromatography, its purity is 99.0%, yield is 92.8%.
Claims (6)
1. from olive growing leaves extract oleanolic acid method it is characterised in that: comprise the following steps:
1), after olive growing leaves being pulverized, extracted with the ethanol solution that volumetric concentration is 70～90%, extract is filtered, takes filtrate
Decompression recycling ethanol, obtains crude extract；
2) crude extract addition volumetric concentration is dissolved again for 50～65% ethanol solutions, obtain mixed solution；
3) by ab-8 large pore resin absorption column on mixed solution, first wash with water to colourless, then adopt 40～50 DEG C, 0.1～
The sodium bicarbonate solution of 0.15mol/l is eluted, then is washed with the sodium bicarbonate solution of 68～72 DEG C, 1～1.5mol/l
De-, collect third time eluent, adjust ph to 3～5, filter, take filter residue and drying, obtain final product.
2. according to claim 1 from olive growing leaves extract oleanolic acid method it is characterised in that: step 1) in,
Olive growing leaves are 1g:10～20ml with the solid-liquid ratio of ethanol solution.
3. according to claim 1 from olive growing leaves extract oleanolic acid method it is characterised in that: step 1) in,
Described extraction time 1～3 time, each extraction time is 1～3h.
4. according to claim 1 from olive growing leaves extract oleanolic acid method it is characterised in that: step 2) in,
Ethanol solution to crude extract concentration is added to be 0.06～0.10g/ml.
5. according to claim 1 from olive growing leaves extract oleanolic acid method it is characterised in that: step 3) in,
Concentration is the elution speed of the sodium bicarbonate solution of 0.1～0.15mol/l is 1～1.5bv/h, and its consumption is 1～1.5bv.
6. according to claim 1 from olive growing leaves extract oleanolic acid method it is characterised in that: step 3) in,
Concentration is the elution speed of the sodium bicarbonate solution of 1～1.5mol/l is 2～2.5bv/h, and its consumption is 0.5～1bv.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610728088.8A CN106336440B (en) | 2016-08-25 | 2016-08-25 | The method of extraction separation oleanolic acid from olive growing leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610728088.8A CN106336440B (en) | 2016-08-25 | 2016-08-25 | The method of extraction separation oleanolic acid from olive growing leaves |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106336440A true CN106336440A (en) | 2017-01-18 |
| CN106336440B CN106336440B (en) | 2018-04-10 |
Family
ID=57822157
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610728088.8A Active CN106336440B (en) | 2016-08-25 | 2016-08-25 | The method of extraction separation oleanolic acid from olive growing leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106336440B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111303236A (en) * | 2020-03-02 | 2020-06-19 | 陕西富恒生物科技有限公司 | Method for simultaneously extracting and separating maslinic acid, oleuropein and oleanolic acid from olive leaves |
| CN113527402A (en) * | 2021-06-04 | 2021-10-22 | 湖南朗林生物资源股份有限公司 | Method for simultaneously extracting oleuropein, maslinic acid and oleanolic acid from olive leaves |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102370713A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Preparation method for momordin |
| CN103169771A (en) * | 2011-12-23 | 2013-06-26 | 中国科学院兰州化学物理研究所 | Method for extracting maslinic acid and oleanolic acid containing mixture from olea europaea L. pomace |
| CN104876998A (en) * | 2015-05-28 | 2015-09-02 | 天津大学 | Method for adopting pH gradient elution method to synchronously separate triterpenoid saponins and licorice flavonoid |
-
2016
- 2016-08-25 CN CN201610728088.8A patent/CN106336440B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102370713A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Preparation method for momordin |
| CN103169771A (en) * | 2011-12-23 | 2013-06-26 | 中国科学院兰州化学物理研究所 | Method for extracting maslinic acid and oleanolic acid containing mixture from olea europaea L. pomace |
| CN104876998A (en) * | 2015-05-28 | 2015-09-02 | 天津大学 | Method for adopting pH gradient elution method to synchronously separate triterpenoid saponins and licorice flavonoid |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111303236A (en) * | 2020-03-02 | 2020-06-19 | 陕西富恒生物科技有限公司 | Method for simultaneously extracting and separating maslinic acid, oleuropein and oleanolic acid from olive leaves |
| CN111303236B (en) * | 2020-03-02 | 2022-10-04 | 陕西富恒生物科技有限公司 | Method for simultaneously extracting and separating maslinic acid, oleuropein and oleanolic acid from olive leaves |
| CN113527402A (en) * | 2021-06-04 | 2021-10-22 | 湖南朗林生物资源股份有限公司 | Method for simultaneously extracting oleuropein, maslinic acid and oleanolic acid from olive leaves |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106336440B (en) | 2018-04-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2020119001A1 (en) | Method for preparing cannabidiol by means of high-speed countercurrent chromatography separation and purification | |
| CN106349324B (en) | The method of extraction separation crataegolic acid from olive growing leaves | |
| CN106967137B (en) | Method for separating high-purity oleuropein by liquid chromatography through macroporous resin combined preparation | |
| CN101229207B (en) | Decoloring refined technology of notoginseng total saponin duolite method | |
| CN103467540A (en) | Method for extracting salidroside from rhodiola | |
| CN105481711B (en) | Hydroxy-beta-sanshool monomer preparation method | |
| CN112321450B (en) | Preparation method of hydroxyl-alpha-sanshool monomer | |
| CN104262446B (en) | The method of mogroside Ⅴ is extracted from Grosvenor Momordica | |
| CN109879919B (en) | Method for separating and preparing three flavonoid glycosides from spina date seeds | |
| CN105713053B (en) | A kind of glucosinolate extraction separation method | |
| CN107522761A (en) | A kind of method and its hypoglycemic purposes for isolating and purifying the O mulberry cloth disaccharide glycosides of delphinidin 3 | |
| CN106336440B (en) | The method of extraction separation oleanolic acid from olive growing leaves | |
| CN104644808A (en) | Process technology of extracting crocin-type component from gardenia | |
| CN105175426B (en) | A kind of method of the extraction purification Bergenin from treebine stem | |
| CN106397529B (en) | The method of extraction separation crataegolic acid from olive pomace | |
| CN101991635A (en) | Preparation method of chickpea total saponin | |
| CN101823964A (en) | Technology for preparing chlorogenic acid in viburnum sargentii koehne leaves | |
| CN109021042B (en) | Method for extracting high-purity oleuropein from olive leaves | |
| CN107213180B (en) | Separation and extraction method of notoginseng flavone | |
| CN101987856A (en) | Preparation method of anthocyanin monomer cornflower-3-glucoside | |
| CN102670935B (en) | Method for extracting total saponins from allium chinense | |
| CN106349321B (en) | The method of extraction separation oleanolic acid from olive pomace | |
| CN108997359A (en) | A method of chlorophyll is extracted from stevioside production waste residue | |
| CN105418572B (en) | It is extracted using microwave counter current, the method for macroporous absorbent resin adsorption and purification whortle anthocyanidin | |
| CN106349325B (en) | The method that separating ursolic acid is extracted from olive growing leaves |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |