CN106349321B - The method of extraction separation oleanolic acid from olive pomace - Google Patents
The method of extraction separation oleanolic acid from olive pomace Download PDFInfo
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- CN106349321B CN106349321B CN201610726918.3A CN201610726918A CN106349321B CN 106349321 B CN106349321 B CN 106349321B CN 201610726918 A CN201610726918 A CN 201610726918A CN 106349321 B CN106349321 B CN 106349321B
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- 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 kind of method that oleanolic acid is extracted from olive pomace, comprise the following steps：1) ethanol solution that olive pomace volumetric concentration is 70~90% is extracted, extract solution is filtered, takes decompression filtrate recycling ethanol, obtains crude extract；2) it is that 50~65% ethanol solutions dissolve again crude extract to be added into volumetric concentration, obtains mixed solution；3) by the large pore resin absorption columns of AB on mixed solution 8, first it is eluted with water to colourless, then eluted using 40~50 DEG C, 0.1~0.15mol/L sodium bicarbonate solution, eluted again with the sodium bicarbonate solution of 68~72 DEG C, 1~1.5mol/L, third time eluent is collected, adjusts pH to 3~5, filtering, filter residue and drying is taken, is produced.The oleanolic acid purity that the present invention extracts is high, and yield is big.
Description
Technical field
The invention belongs to technical field of biological extraction, and in particular to one kind extraction separation oleanolic acid from olive pomace
Method.
Background technology
Olive (Olea europare L.) is world-renowned woody oil tree species, and the cultivation for having more than 4000 years is gone through
History.Olea europaea fruit produces substantial amounts of pomace after squeezing, most of directly to go out of use, and not only seriously pollutes environment, also result in
The waste of resource.Olive pomace is made up of olive pericarp, fruit stone and remaining pulp, wherein mainly containing triterpenes, flavones
The active ingredients such as class, volatile oil, vitamin.Triterpene compound is mainly made up of 6 isoprene structures units, most of
For the terpenoid containing 30 carbon atoms.Olive pomace triterpenes components mainly include oleanolic acid, crataegolic acid and black bearberry
Acid, and these three structures of matter are similar, generally use Column chromatography techniques are separated, but repeatedly silica gel column chromatography separation and
Recrystallization causes the sample size that finally gives seldom.Again due to Column chromatography techniques generally existing long processing period, continuous operation property
It is not strong, it is not easy the industrial production process for being used for bioactive substance as separating medium.
The content of the invention
Present invention solves the technical problem that it is to provide a kind of method that oleanolic acid is extracted from olive pomace, this method
Simply, the cycle is short, and obtained oleanolic acid purity is high, and yield is big.
Technical scheme provided by the invention is the method that oleanolic acid is extracted from olive pomace, is comprised the following steps：
1) ethanol solution that olive pomace volumetric concentration is 70~90% is extracted, extract solution is filtered, takes filtrate
Ethanol is recovered under reduced pressure, obtains crude extract；
2) it is that 50~65% ethanol solutions dissolve again crude extract to be added into volumetric concentration, obtains mixed solution；
3) by AB-8 large pore resin absorption columns on mixed solution, be first eluted with water to colourless, then using 40~50 DEG C,
0.1~0.15mol/L sodium bicarbonate solution is eluted, then is entered with the sodium bicarbonate solution of 68~72 DEG C, 1~1.5mol/L
Row elution, collects third time eluent, adjusts pH to 3~5, filtering, takes filter residue and drying, produce.
In step 1), use volumetric concentration to extract olive pomace for 70~90% ethanol solution, can fully extract
Oleanolic acid, crataegolic acid and ursolic acid in olive pomace.The solid-liquid ratio of olive pomace and ethanol solution is 1g：10~
20ml.Described to be extracted as heating and refluxing extraction, extraction time is 1~3 time, and each extraction time is 1~3h.
In step 2), use volumetric concentration to dissolve crude extract for 50~65% ethanol solution, can remove pole in crude extract
The less impurity of property, play the effect of preliminary purification.It is 0.06~0.10g/ml to add ethanol solution to crude extract concentration, now
The adsorption effect of upper prop is optimal.
In step 3), the macroreticular resins of AB-8 models is low pole, and average pore size is 13~14nm, specific surface area 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 adsorbed, when resin aperture be less than absorption it is effective into
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 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 preferable using the adsorption effect of AB-8 macroreticular resins.Resin
The blade diameter length ratio of post is 1:3~6 are advisable, with 1:4 is optimal.
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 are difficult to be isolated to come, but the present invention uses 40~50 DEG C, 0.1~0.15mol/L bicarbonate
Sodium solution can be larger with polarity in resin hawthorn acid reaction generation soluble sodium salt, and then elute, and polarity is less than normal
Oleanolic acid and the carboxyl of ursolic acid and the polarizability of hydrone it is poor, it is impossible to ionize out H+, can not enter with sodium acid carbonate
Row reaction generation sodium salt, then stay in resin column.The elution speed of sodium bicarbonate solution is 1~1.5BV/h, its dosage is 1~
During 1.5BV, elution effect is preferable.
Eluted using the sodium bicarbonate solution of 68~72 DEG C, 1~1.5mol/L, can now elute oleanolic acid,
And ursolic acid is not eluted, because oleanolic acid and ursolic acid structure are isomer, therefore, using faster elution speed
2~2.5BV/h, less amount of eluent are 0.5~1BV, can at utmost elute oleanolic acid, and ursolic acid is not eluted.
Method of the invention is simple, the cycle is short, and the oleanolic acid purity of extraction is high, and yield is big.
Embodiment
The present invention is further elaborated for specific examples below, but not as a limitation of the invention.
Embodiment 1
1) it is 1g by solid-liquid ratio by ethanol solution that olive pomace and volumetric concentration are 70%：10ml ratio mixing,
Heating and refluxing extraction 3h, extract solution is filtered, and takes decompression filtrate recycling ethanol, obtains crude extract；
2) it is that 50% ethanol solution to crude extract concentration is 0.06g/ml by crude extract addition volumetric concentration, stirs evenly, obtain
Mixed solution；
3) by AB-8 large pore resin absorption columns on mixed solution, resin column blade diameter length ratio is 1:3, first it is eluted with water to colourless,
Then 40 DEG C of addition, 0.1mol/L sodium bicarbonate solution are eluted, and the elution speed of sodium bicarbonate solution is 1BV/h, its
Dosage is 1BV, then is eluted with the sodium bicarbonate solution of 68 DEG C, 1mol/L, elution speed 2BV/h, and its dosage is
0.5BV, third time eluent (i.e. the sodium acid carbonate eluent of 1mol/L) is collected, adjust pH to 3, filtering, take filter residue and drying, i.e.,
.
It is measured according to high performance liquid chromatography, its purity is 99.1%, yield 93.3%.
Embodiment 2
1) it is 1g by solid-liquid ratio by ethanol solution that olive pomace and volumetric concentration are 90%：20ml ratio mixing,
Heating and refluxing extraction 3 times, each extraction time are 3h, and extract solution is filtered, takes decompression filtrate recycling ethanol, obtains crude extract；
2) it is that 65% ethanol solution to crude extract concentration is 0.10g/ml by crude extract addition volumetric concentration, stirs evenly, obtain
Mixed solution；
3) by AB-8 large pore resin absorption columns on mixed solution, resin column blade diameter length ratio is 1:6, first it is eluted with water to colourless,
Then 50 DEG C of addition, 0.15mol/L sodium bicarbonate solution are eluted, and the elution speed of sodium bicarbonate solution is 1.5BV/h,
Its dosage is 1.5BV, then is eluted with the sodium bicarbonate solution of 72 DEG C, 1.5mol/L, elution speed 2.5BV/h, and it is used
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
It is dry, produce.
It is measured according to high performance liquid chromatography, its purity is 99.0%, yield 93.0%.
Embodiment 3
1) it is 1g by solid-liquid ratio by ethanol solution that olive pomace and volumetric concentration are 90%：20ml ratio mixing,
Heating and refluxing extraction 2 times, each extraction time are 2h, and extract solution is filtered, takes decompression filtrate recycling ethanol, obtains crude extract；
2) it is that 55% ethanol solution to crude extract concentration is 0.08g/ml by crude extract addition volumetric concentration, stirs evenly, obtain
Mixed solution；
3) by AB-8 large pore resin absorption columns on mixed solution, resin column blade diameter length ratio is 1:4, first it is eluted with water to colourless,
Then 45 DEG C of addition, 0.12mol/L sodium bicarbonate solution are eluted, and the elution speed of sodium bicarbonate solution is 1.2BV/h,
Its dosage is 1.2BV, then is eluted with the sodium bicarbonate solution of 70 DEG C, 1.35mol/L, elution speed 2.5BV/h, and it is used
Measure as 0.5BV, collect third time eluent (1.35mol/L sodium acid carbonate eluent), adjust pH to 4, filtering, take filter residue to do
It is dry, produce.
It is measured according to high performance liquid chromatography, its purity is 99.5%, yield 94.5%.
Embodiment 4
1) it is 1g by solid-liquid ratio by ethanol solution that olive pomace and volumetric concentration are 70%：20ml ratio mixing,
Heating and refluxing extraction 3 times, each extraction time are 1h, and extract solution is filtered, takes decompression filtrate recycling ethanol, obtains crude extract；
2) it is that 65% ethanol solution to crude extract concentration is 0.06g/ml by crude extract addition volumetric concentration, stirs evenly, obtain
Mixed solution；
3) by AB-8 large pore resin absorption columns on mixed solution, resin column blade diameter length ratio is 1:3, first it is eluted with water to colourless,
Then 50 DEG C of addition, 0.1mol/L sodium bicarbonate solution are eluted, and the elution speed of sodium bicarbonate solution is 1.5BV/h,
Its dosage is 1BV, then is eluted with the sodium bicarbonate solution of 68 DEG C, 1.5mol/L, elution speed 2BV/h, and its dosage is
1BV, third time eluent (i.e. the sodium acid carbonate eluent of 1.5mol/L) is collected, adjust pH to 3~5, filtering, take filter residue to do
It is dry, produce.
It is measured according to high performance liquid chromatography, its purity is 99.0%, yield 92.8%.
Claims (6)
1. the method for oleanolic acid is extracted from olive pomace, it is characterised in that：Comprise the following steps：
1) ethanol solution that olive pomace volumetric concentration is 70~90% is extracted, extract solution is filtered, takes filtrate decompression
Ethanol is reclaimed, obtains crude extract；
2) it is that 50~65% ethanol solutions dissolve again crude extract to be added into volumetric concentration, obtains mixed solution；
3) by AB-8 large pore resin absorption columns on mixed solution, be first eluted with water to colourless, then using 40~50 DEG C, 0.1~
0.15mol/L sodium bicarbonate solution is eluted, then is washed with the sodium bicarbonate solution of 68~72 DEG C, 1~1.5mol/L
It is de-, third time eluent is collected, pH to 3~5 is adjusted, filtering, takes filter residue and drying, produce.
2. the method according to claim 1 that oleanolic acid is extracted from olive pomace, it is characterised in that：Step 1)
In, the solid-liquid ratio of olive pomace and ethanol solution is 1g：10~20ml.
3. the method according to claim 1 that oleanolic acid is extracted from olive pomace, it is characterised in that：Step 1)
In, the extraction time 1~3 time, each extraction time is 1~3h.
4. the method according to claim 1 that oleanolic acid is extracted from olive pomace, it is characterised in that：Step 2)
In, it is 0.06~0.10g/ml to add ethanol solution to crude extract concentration.
5. the method according to claim 1 that oleanolic acid is extracted from olive pomace, it is characterised in that：Step 3)
In, the elution speed for the sodium bicarbonate solution that concentration is 0.1~0.15mol/L is 1~1.5BV/h, and its dosage is 1~1.5BV.
6. the method according to claim 1 that oleanolic acid is extracted from olive pomace, it is characterised in that：Step 3)
In, the elution speed for the sodium bicarbonate solution that concentration is 1~1.5mol/L is 2~2.5BV/h, and its dosage is 0.5~1BV.
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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 |
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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 |
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Effective date of registration: 20180905 Address after: 362199 Room 401, 50 Jianan Road, Huian Town, Fujian Province, 401 Patentee after: Zhuang Mingchuan Address before: 541004 B303, room B, business building, five building, District five, New District, Guilin, Qixing District, the Guangxi Zhuang Autonomous Region. Patentee before: Guilin Yi Tiancheng Biotechnology Co. Ltd. |
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Effective date of registration: 20190111 Address after: 530007 Tianyu Huating Building 1001, 1002, 29 Kede West Road, Nanning City, Guangxi Zhuang Autonomous Region Patentee after: Guangxi Huikang biological Polytron Technologies Inc Address before: 362199 Room 401, 50 Jianan Road, Huian Town, Fujian Province, 401 Patentee before: Zhuang Mingchuan |