CN106349324A - Method for extracting and separating maslinic acid from olive leaves - Google Patents
Method for extracting and separating maslinic acid from olive leaves Download PDFInfo
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- CN106349324A CN106349324A CN201610728726.6A CN201610728726A CN106349324A CN 106349324 A CN106349324 A CN 106349324A CN 201610728726 A CN201610728726 A CN 201610728726A CN 106349324 A CN106349324 A CN 106349324A
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- oleae europaeae
- fructus oleae
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
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Abstract
The invention discloses a method for extracting maslinic acid from olive leaves. The method comprises the following steps: 1) extracting olive leaves with a 70-90 vol% ethanol solution, filtering the extracting solution, taking the filtrate, and recovering the ethanol under reduced pressure to obtain a crude extract; 2) adding the crude extract into a 50-65 vol% ethanol solution to be redissolved, thereby obtaining a mixed solution; and 3) passing the mixed solution through an AB-8 macroporous adsorbent resin column, eluting with water to a colorless state, eluting with a 40-50-DEG C 0.1-0.15 mol/L sodium bicarbonate solution, collecting the sodium bicarbonate eluate, regulating the pH value to 3-5, filtering, taking the filter residue, and drying. The maslinic acid extracted by the method has the advantages of high purity and high yield.
Description
Technical field
The invention belongs to technical field of biological extraction is and in particular to a kind of extract the side separating Crataegolic acid from leaf of Fructus oleae europaeae
Method.
Background technology
Fructus oleae europaeae (olea europare l.) is world-renowned woody oil tree species, has the cultivation of more than 4000 year to go through
History.The decoct of leaf of Fructus oleae europaeae or preserved material have certain curative effect in mediterranean country frequently as the medication among the people of diabetes.Fructus oleae europaeae
Leaf extract is used as dietary supplement to strengthen immunologic function in America and Europe.Modern data shows, contains in leaf of Fructus oleae europaeae
The effective ingredient such as flavonoid, triterpeness, saccharide, row iridoidses.Triterpenoid compound is mainly by 6 isoprene structures
Unit is constituted, and major part is the terpenoid containing 30 carbon atoms.In leaf of Fructus oleae europaeae, 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 commercial production process of bioactive substance as separating medium.
Content of the invention
Present invention solves the technical problem that being to provide a kind of method extracting Crataegolic acid from leaf of Fructus oleae europaeae, the method letter
Single, cycle is short, the Crataegolic acid purity obtaining is high, and yield is big.
The technical scheme that the present invention provides is the method extracting Crataegolic acid from leaf of Fructus oleae europaeae, comprises the following steps:
1) leaf of Fructus oleae europaeae is pulverized, extracted with the ethanol solution that volumetric concentration is 70～90%, extracting solution is filtered, takes filter
Liquid decompression recycling ethanol, obtains crude extract；
2) crude extract addition volumetric concentration is dissolved again for 50～65% ethanol solution, obtain mixed solution；
3) by ab-8 macroporous adsorptive resins on mixed solution, first wash with water to colourless, be subsequently adding 40～50 DEG C,
The sodium bicarbonate solution of 0.1～0.15mol/l carries out eluting, collects sodium bicarbonate eluent, adjusts ph to 3～5, filters, takes
Filter residue and drying, obtains final product.
In step 1) in, leaf of Fructus oleae europaeae is extracted using the ethanol solution that volumetric concentration is 70～90%, can abundant extract oil
Oleanolic acid in Folium olive, Crataegolic acid and ursolic acid.Leaf of Fructus oleae europaeae is 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 macroporous 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 absorption with macroporous adsorbent resin material, 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, effective ingredient molecule can not enter inside resin, can only adsorb in resin outer surface, when resin aperture is more than effectively
During component molecules, effective ingredient can adsorb and also 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 macroporous 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 Fructus Crataegi 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 bicarbonate
Row reaction generates sodium salt, then stay in resin column.
The method of the present invention is simple, cycle is short, and the Crataegolic 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) leaf of Fructus oleae europaeae is crushed to 100 mesh, the ethanol solution being 70% with volumetric concentration is by solid-liquid ratio for 1g:10ml's
Ratio mixes, and extracting solution 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 macroporous adsorptive resins 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 carry out eluting, the elution speed of sodium bicarbonate solution is 1bv/h, its
Consumption is 1bv, collects sodium bicarbonate eluent, adjusts ph to 3, filters, takes filter residue and drying, obtain final product.According to high performance liquid chromatography
Method is measured, and its purity is 99.1%, and yield is 95.3%.
Embodiment 2
1) leaf of Fructus oleae europaeae is 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, extracting solution 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 macroporous adsorptive resins 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 carry out eluting, the elution speed of sodium bicarbonate solution is 1.5bv/h,
Its consumption is 1.5bv, collects sodium bicarbonate eluent, adjusts ph to 5, filters, takes filter residue and drying, obtain final product.According to efficient liquid phase
Chromatography is measured, and its purity is 99.0%, and yield is 95.1%.
Embodiment 3
1) leaf of Fructus oleae europaeae is crushed to 200 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, extracting solution 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 macroporous adsorptive resins 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 carry out eluting, the elution speed of sodium bicarbonate solution is 1.2bv/h,
Its consumption is 1.2bv, collects sodium bicarbonate eluent, adjusts ph to 4, filters, takes filter residue and drying, obtain final product.According to efficient liquid phase
Chromatography is measured, and its purity is 99.5%, and yield is 96.8%.
Embodiment 4
1) leaf of Fructus oleae europaeae is crushed to 150 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, extracting solution 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 macroporous adsorptive resins 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 carry out eluting, the elution speed of sodium bicarbonate solution is 1.5bv/h,
Its consumption is 1bv, collects sodium bicarbonate eluent, adjusts ph to 5, filters, takes filter residue and drying, obtain final product.According to high-efficient liquid phase color
Spectrometry is measured, and its purity is 99.0%, and yield is 95.4%.
Claims (5)
1. from leaf of Fructus oleae europaeae extract Crataegolic acid method it is characterised in that: comprise the following steps:
1) leaf of Fructus oleae europaeae is pulverized, extracted with the ethanol solution that volumetric concentration is 70～90%, extracting solution is filtered, takes filtrate to subtract
Push back receipts ethanol, obtain crude extract；
2) crude extract addition volumetric concentration is dissolved again for 50～65% ethanol solution, obtain mixed solution；
3) by ab-8 macroporous adsorptive resins on mixed solution, first wash with water to colourless, be subsequently adding 40～50 DEG C, 0.1～
The sodium bicarbonate solution of 0.15mol/l carries out eluting, collects sodium bicarbonate eluent, adjusts ph to 3～5, filters, and takes filtering residue to do
Dry, obtain final product.
2. according to claim 1 from leaf of Fructus oleae europaeae extract Crataegolic acid method it is characterised in that: step 1) in, oil
Folium olive is 1g:10～20ml with the solid-liquid ratio of ethanol solution.
3. according to claim 1 from leaf of Fructus oleae europaeae extract Crataegolic acid method it is characterised in that: step 1) in, institute
State extraction time 1～3 time, each extraction time is 1～3h.
4. according to claim 1 from leaf of Fructus oleae europaeae extract Crataegolic acid method it is characterised in that: step 2) in, plus
Entering ethanol solution to crude extract concentration is 0.06～0.10g/ml.
5. according to claim 1 from leaf of Fructus oleae europaeae extract Crataegolic acid method it is characterised in that: step 3) in, carbon
The elution speed of sour hydrogen sodium solution is 1～1.5bv/h, and its consumption is 1～1.5bv.
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| CN201610728726.6A CN106349324B (en) | 2016-08-25 | 2016-08-25 | The method of extraction separation crataegolic acid from olive growing leaves |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109371094A (en) * | 2018-12-19 | 2019-02-22 | 湖南杰萃生物技术有限公司 | A method of extracting dehydrogenation hawthorn acid from olive leaf |
| 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 |
| CN112409437A (en) * | 2020-12-12 | 2021-02-26 | 南京康齐生物科技有限公司 | Method for extracting and separating high-purity maslinic 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 |
| CN113952269A (en) * | 2021-12-10 | 2022-01-21 | 东莞波顿香料有限公司 | Preparation method and application of olive leaf extract |
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| 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 |
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2016
- 2016-08-25 CN CN201610728726.6A patent/CN106349324B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109371094A (en) * | 2018-12-19 | 2019-02-22 | 湖南杰萃生物技术有限公司 | A method of extracting dehydrogenation hawthorn acid from olive leaf |
| 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 |
| CN112409437A (en) * | 2020-12-12 | 2021-02-26 | 南京康齐生物科技有限公司 | Method for extracting and separating high-purity maslinic 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 |
| CN113952269A (en) * | 2021-12-10 | 2022-01-21 | 东莞波顿香料有限公司 | Preparation method and application of olive leaf extract |
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