CN1552722A - Method for extracting and separating oleanolic acid - Google Patents
Method for extracting and separating oleanolic acid Download PDFInfo
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- CN1552722A CN1552722A CNA031366880A CN03136688A CN1552722A CN 1552722 A CN1552722 A CN 1552722A CN A031366880 A CNA031366880 A CN A031366880A CN 03136688 A CN03136688 A CN 03136688A CN 1552722 A CN1552722 A CN 1552722A
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- oleanolic acid
- buckthorn
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Abstract
A process for extracting oleanolic acid from fruit, seed and leaf of seabuthorn through drying, pulverizing, supercritical CO2 extracting, reflux extracting of dregs three times, centrifugal separating, collecting the liquid extract, filtering by 200-300 meshes and then by 400-500 meshes, membrane filtering twice at 40-60 deg.C under 0.05-0.20 MPa, and vacuum (or spray) drying.
Description
Affiliated field
The present invention relates to the extraction preparation method of effective ingredients in plant, specifically the method for extraction separation Oleanolic Acid from plant.
Technical background
Oleanolic Acid (oleanolic acid, have another name called celebrating four elements) belongs to pentacyclic triterpenoid, is present in the various plants with the form of free daughter and glycoside.Foreign literature report Oleanolic Acid has anti-inflammatory, calmness, cardiac stimulant, diuresis, enhancing immunity and inhibition S
180Effects such as knurl strain growth.Experiment shows that Oleanolic Acid is to CCl
4The acute and chronic liver injury that causes has significant protective effect; can prevent the generation of experimental liver cirrhosis; and by suppressing or alleviate the necrosis of hepatocellular degeneration; promote liver cell regeneration; quicken the reparation of necrotic tissue; suppress the reaction of collegen filament hyperplasia and specificity inflammation,, bring into play its hepatoprotective effect so that liver cirrhosis is difficult to form.In vitro tests proves that also Oleanolic Acid has tangible antivirus action.The active functional group of Oleanolic Acid makes it competitively with the growth of hepatitis virus and breeds necessary enzyme to combine, thereby suppresses hepatitis B virus.The clinical Oleanolic Acid that is used for acute icterohepatitis, chronic hepatitis mainly is to extract from plants such as Glossy Privet Fruit, Mile Swertia Herbs.Also have report from
Extraction separation in wood, the pawpaw.
The extraction separation of Oleanolic Acid adopts the method for alcohol extracting, chromatographic separation more at present.For example: " Chinese wild plant resource " the 20th the 2nd phase of volume 48 pages disclosed " extraction and separation method of Oleanolic Acid and assay in the pawpaw " is exactly with an amount of 95% alcohol immersion, diacolation, suction filtration with the pawpaw raw material, after must soaking paste, add anhydrous alcohol solution, add the gac reflux decolour, remove behind the gac reconcentration to a certain amount of with suction method, add an amount of silica gel for chromatography and adsorb, mix thoroughly, grind into fine powder dress post, ethyl ester one sherwood oil with certain ratio is that elutriant carries out wash-out successively, obtains Oleanolic Acid in the 25-42 cut.The weak point that this method exists is: process cycle is long, efficient is low, the effective constituent consume is big, and will consume a large amount of organic solvents in sepn process, has so not only strengthened the production cost of Oleanolic Acid, returns environment protection and causes certain pressure.
Summary of the invention
The purpose of this invention is to provide a kind of method of new extraction separation Oleanolic Acid, this method technology is simple, cost is low, Oleanolic Acid enriching quantity height and ecotope do not had negative impact.
The object of the present invention is achieved like this: the method for the extraction separation Oleanolic Acid that is provided may further comprise the steps:
A, be raw material with Fructus Hippophae, seabuckthorn fruit peel slag, sea-buckthorn seed or Folium Hippophae, negative pressure drying makes its moisture≤7% respectively, and pulverizing is 40～80 purpose fine powders;
B, with the fine powder of Fructus Hippophae, seabuckthorn fruit peel slag, sea-buckthorn seed, adopt supercritical CO
2Extraction, seabuckthorn fruit oil, Fructus Hippophae dreg oil, the Seabuckthorm Seed Oil that extraction obtains done him in addition and used (these extracts have the pharmaceutical use of himself).Leave and take the full pomace of extracted residual, peel residue, seed slag;
C, the full pomace with leaving and taking in Folium Hippophae in a step or the b step, peel residue, seed slag add 6～8 times, 60～90% edible ethanol, refluxing extraction 2-3 time, and centrifugal, united extraction liquid obtains sea-buckthorn Oleanolic Acid extracting solution;
D, sea-buckthorn Oleanolic Acid extracting solution is first with 200～300 purpose flame filter press coarse filtration with the smart filter of 400～500 purpose flame filter presses, gets smart filtrate again;
E, smart filtrate are that with the membrane filtration of molecular weight cut-off 1500～2000dalton, filtered liquid is used the membrane filtration of molecular weight cut-off 300dalton again, obtains sea-buckthorn Oleanolic Acid concentrated solution earlier under the condition of 40～60 ℃ of 0.10～0.50Mpa, temperature at pressure;
F, with sea-buckthorn Oleanolic Acid concentrated solution in drying under reduced pressure or spraying drying below 60 ℃, sea-buckthorn Oleanolic Acid powder.
The pressure preferable range is 0.20～0.40Mpa during membrane filtration of the present invention.
Innovation part of the present invention is to adopt membrane technique extraction separation from Fructus Hippophae, seabuckthorn fruit peel slag, sea-buckthorn seed and Folium Hippophae to go out Oleanolic Acid.It provides a kind of extraction and separation method of new Oleanolic Acid for people.
Sea-buckthorn Oleanolic Acid molecular weight is 456.71, and the oleanolic acid glucoside molecular weight is generally about 900.The present invention adopts membrane separation technique, the material in this molecular weight ranges in the seabuckthorn plant is carried out extraction separation, its extract is carried out qualitative and quantitative analysis, determine that it contains a large amount of Oleanolic Acids, thereby, innovation part of the present invention also the has been effective enrichment method Oleanolic Acid in the seabuckthorn plant.
The Oleanolic Acid of the inventive method extraction separation adopts 2000 editions Pharmacopoeias of People's Republic of China (an one) 34 pages, and tlc is carried out qualitative analysis.
Its quantitative analysis method is:
Get the sea-buckthorn Oleanolic Acid powder 1g of the inventive method extraction separation, 3 times (20ml, 15ml 10ml), filter with the dehydrated alcohol refluxing extraction.Be settled to 50ml, with millipore membrane filtration sample introduction.Chromatographic column: C
18-ODS post, moving phase: methanol-water (88: 12), detect wavelength 220nm, sensitivity 0.05AUFS, flow velocity 0.6ml/ minute, room temperature.External standard method is (" herbal medicine " 1980.11 (6) 385 pages such as see that poplar is just being weighed quantitatively.
Because the inventive method is to adopt membrane separation technique that the material in the certain molecular weight scope is separated, thereby pentacyclic triterpenoid such as the ursolic acid suitable with the Oleanolic Acid molecular weight can propose in the lump.
The inventive method is compared with traditional Oleanolic Acid extracting method, and it is simple also to have technology, easy to operate, and the separation efficiency height is pollution-free, no solvent residue, the characteristics that cost is low.
The medicine that can be used for preparing treatment hepatitis, liver cirrhosis with the Oleanolic Acid of the inventive method preparation.Also can be used for the medicine that preparation conforms to its pharmacological action (as anti-inflammatory, calmness, cardiac stimulant, diuresis, enhancing immunity).
Embodiment
Embodiment 1:
Take by weighing seabuckthorn fruit peel slag 10kg, negative pressure drying makes its moisture≤7%, pulverizes, and crosses 80 mesh sieves, with SFE～24L supercritical CO
2The device extraction, at pressure 28Mpa, 45 ℃ of temperature, 200 liters/hour of flows extract under the 2.5 hours time condition.The Fructus Hippophae dreg oil that extraction obtains is made his usefulness in addition, removes remaining peel residue (9.6kg) behind the Fructus Hippophae dreg oil, adds 8 times of 70% edible ethanol refluxing extraction 2 times, centrifugal, united extraction liquid, extracting solution earlier with 200 keevil frame filter coarse filtration, with the smart filter of 500 keevil frame filters, is got smart filtrate again.Smart filtrate is carried out membrane sepn with UF2540 ultra-filtration membrane device, at pressure: 0.30～0.50Mpa, under the temperature 45-55 ℃ of condition, use the membrane filtration of molecular weight cut-off 1500dalton earlier, remove the material of the macromole (as soluble proteins, larger molecular organics etc.) that contains in the solution, obtain sea-buckthorn Oleanolic Acid filtrate, use the membrane filtration of molecular weight cut-off 300dalton again, remove small molecules in the solution (as inorganicss such as iron, calcium, sodium, magnesium and molecular weight at the material below 300), remove ethanol and moisture simultaneously, get sea-buckthorn Oleanolic Acid concentrated solution.With sea-buckthorn Oleanolic Acid concentrated solution drying under reduced pressure (below 60 ℃), promptly get sea-buckthorn Oleanolic Acid isolate 98.55g, the content 28.5% of its sea-buckthorn Oleanolic Acid again.
Embodiment 2:
Take by weighing Folium Hippophae 10kg, negative pressure drying makes its moisture≤7%, pulverize, cross 60 mesh sieves, get the Folium Hippophae powder, add 8 times of 70% edible ethanol refluxing extraction 3 times, centrifugal, united extraction liquid, extracting solution is carried out coarse filtration with 300 keevil frame filters earlier, carry out the essence filter with 500 keevil frame filters again, get the smart filtrate of Folium Hippophae Oleanolic Acid.Smart filtrate is carried out membrane sepn with the ultra-filtration membrane device, at pressure: 0.10～0.30Mpa, under the temperature 45-55 ℃ of condition, use the membrane filtration of molecular weight cut-off 2000dalaton earlier, remove the macromolecular material in the solution, use the membrane filtration of molecular weight cut-off 300dalton again, remove the micromolecular material in the solution, remove the ethanol and the aqueous solution simultaneously, get sea-buckthorn Oleanolic Acid concentrated solution.With sea-buckthorn Oleanolic Acid concentrated solution spraying drying, promptly get sea-buckthorn Oleanolic Acid isolate 86.5g, the content 20.5% of its sea-buckthorn Oleanolic Acid again.
Claims (2)
1, a kind of method of extraction separation Oleanolic Acid is characterized in that it may further comprise the steps:
A, be raw material with Fructus Hippophae, seabuckthorn fruit peel slag, sea-buckthorn seed or Folium Hippophae, negative pressure drying makes its moisture≤7% respectively, and pulverizing is 40～80 purpose fine powders;
B, with the fine powder of Fructus Hippophae, seabuckthorn fruit peel slag, sea-buckthorn seed, adopt supercritical CO
2The full pomace of extracted residual, peel residue, seed slag are left and taken in extraction;
C, the full pomace with in Folium Hippophae in a step or the b step, peel residue, seed slag really add 6～8 times, 60～90% edible ethanol refluxing extraction 2-3 time, and centrifugal merging obtains sea-buckthorn Oleanolic Acid extracting solution;
D, sea-buckthorn Oleanolic Acid extracting solution is first with 200～300 purpose sheet frame coarse filtration with the smart filter of 400～500 purpose sheet frames, gets smart filtrate again;
E, smart filtrate under the condition of 40～60 ℃ of 0.10～0.50Mpa, temperature, earlier with the membrane filtration of molecular weight cut-off 1500～2000dalton, use the membrane filtration of molecular weight cut-off 300dalton again, obtain sea-buckthorn Oleanolic Acid concentrated solution;
F, with sea-buckthorn Oleanolic Acid concentrated solution in drying under reduced pressure or spraying drying below 60 ℃, sea-buckthorn Oleanolic Acid powder.
2, the method for extraction separation Oleanolic Acid according to claim 1, the pressure that it is characterized in that the membrane filtration in the said e step is 0.20～0.40Mpa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031366880A CN1552722A (en) | 2003-05-26 | 2003-05-26 | Method for extracting and separating oleanolic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA031366880A CN1552722A (en) | 2003-05-26 | 2003-05-26 | Method for extracting and separating oleanolic acid |
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| Publication Number | Publication Date |
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| CN1552722A true CN1552722A (en) | 2004-12-08 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNA031366880A Pending CN1552722A (en) | 2003-05-26 | 2003-05-26 | Method for extracting and separating oleanolic acid |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974059A (en) * | 2010-09-10 | 2011-02-16 | 武汉巨惠绿色产业发展有限公司 | Preparation of oleanolic acid structure analogues and semisynthesis method of oleanolic acid |
| CN101857625B (en) * | 2009-11-18 | 2013-03-27 | 青海清华博众生物技术有限公司 | Method for extracting oleanolic acid from sea-buckthorn |
| CN108434182A (en) * | 2018-04-09 | 2018-08-24 | 内蒙古宇航人高技术产业有限责任公司 | A kind of supercritical CO2The method that fluid extraction technology extracts Leaves of Hippophae L polyphenol |
| CN110317239A (en) * | 2019-08-13 | 2019-10-11 | 南京津奇生物科技有限公司 | A kind of preparation method for extracting Corosolic acid from fructus hippophae |
| CN111004683A (en) * | 2019-12-26 | 2020-04-14 | 贵州中烟工业有限责任公司 | Sea-buckthorn fruit essence, production method of sea-buckthorn fruit essence and tobacco product |
-
2003
- 2003-05-26 CN CNA031366880A patent/CN1552722A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101857625B (en) * | 2009-11-18 | 2013-03-27 | 青海清华博众生物技术有限公司 | Method for extracting oleanolic acid from sea-buckthorn |
| CN101974059A (en) * | 2010-09-10 | 2011-02-16 | 武汉巨惠绿色产业发展有限公司 | Preparation of oleanolic acid structure analogues and semisynthesis method of oleanolic acid |
| CN108434182A (en) * | 2018-04-09 | 2018-08-24 | 内蒙古宇航人高技术产业有限责任公司 | A kind of supercritical CO2The method that fluid extraction technology extracts Leaves of Hippophae L polyphenol |
| CN110317239A (en) * | 2019-08-13 | 2019-10-11 | 南京津奇生物科技有限公司 | A kind of preparation method for extracting Corosolic acid from fructus hippophae |
| CN111004683A (en) * | 2019-12-26 | 2020-04-14 | 贵州中烟工业有限责任公司 | Sea-buckthorn fruit essence, production method of sea-buckthorn fruit essence and tobacco product |
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