CN102532217A - Method for purifying and separating high-content oleuropein from olive leaves - Google Patents
Method for purifying and separating high-content oleuropein from olive leaves Download PDFInfo
- Publication number
- CN102532217A CN102532217A CN2011104391867A CN201110439186A CN102532217A CN 102532217 A CN102532217 A CN 102532217A CN 2011104391867 A CN2011104391867 A CN 2011104391867A CN 201110439186 A CN201110439186 A CN 201110439186A CN 102532217 A CN102532217 A CN 102532217A
- Authority
- CN
- China
- Prior art keywords
- leaf
- content
- cleupin
- fructus oleae
- oleae europaeae
- 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
Landscapes
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a method for purifying and separating high-content oleuropein from olive leaves, which comprises cutting olive leaves, extracting at a low temperature, filtering, concentrating or directly performing chromatography, concentrating at a low temperature, and drying to obtain a product with oleuropein content higher than 80%. The invention adopts a certain concentration of ethanol or water as an extraction solvent for low-temperature extraction, and utilizes resin tandem or secondary adsorption chromatography for separation and purification to obtain the product with oleuropein content higher than 80%. The invention has the advantages of low energy consumption, short production period, environmental friendliness and a simple operation, and is especially suitable for large-scale industrial production.
Description
Technical field
The present invention relates to the method for extraction separation high-content cleupin from leaf of Fructus oleae europaeae.
Background technology
Fructus oleae europaeae belongs to Oleaceae Olea aiphyllium, is that the double fruit of world-renowned woody oleiferous plants uses seeds, Cultivar that higher edibleness is arranged, and contains and enriches kind of high-quality edible plant oil---and Fructus oleae europaeae oil is famous subtropical fruit tree and important commodity trees.Mainly be distributed in European Mediterranean Sea coastwise contries and California, USA area, Greece, Italy, Tunisia, Spain are for concentrating the place of production.The municipalization introducing culture is economized in China western part and Gansu, the Northwest, Shaanxi, Sichuan, Chongqing, Fujian, Guangxi, Guangdong, Hunan etc.The existing Fructus oleae europaeae in the whole nation surpasses 1,600 ten thousand strains.Longnan City, Gansu Province Wudu District is present national maximum oil olive base.
Mainly contain compositions such as secoiridoid and glycosides, flavones and glycosides thereof, bisflavone and glycosides thereof, low branch Chinese tannin in the olive leaf, the secoiridoid class is a main active ingredient.Staple is the pained material of iridoids in the Olive leaf P.E, and what its activity was the highest is cleupin.
Cleupin is a kind of bimolecular structure phenols and Flavonoid substances; Have natural anti-oxidant, antitumor, delay senility, strengthen cardiovascular function, treatment chronic prostatitis, strengthening immunity, conciliation endocrine system, protect important nourishing functions such as liver, anti-inflammatory; Also have reducing blood-fat simultaneously, fall the pharmacological action of blood chaff, be widely used in fields such as food, cosmetic and bio-pharmaceuticals.
High-load cleupin is mainly used in skin care item, can protect skin cells not receive ultraviolet injury, effectively keeps the tender and elasticity of skin, reaches the effect of holding up skin, tender skin.Microherb high-content cleupin 80% is special in skin care item design, and its effective constituent is high, and is of light color, is fit to very much the cosmetic formulations design.
The olive leaf extractive technique mainly includes machine solvent extraction techniques, supercritical CO 2 abstraction technique, ultrasonic extraction technology and microwave-assisted extraction technique etc.Organic solvent extraction has shortcomings such as foreign matter content height, solvent consumption is big, cost is high, pollution is big.Supercritical CO 2 extraction has advantages such as the content of extraction is high, pollution-free, but have high input, the cost height.Though the ultrasonic microwave extracting that involves has advantages such as energy-conservation, efficient, on producing, be difficult to realize change on a large scale application.
Bibliographical information or the disclosed cleupin extraction and separation technology of patent are more at present, but cleupin content reaches the rare report of separating and purifying technology more than 80%.The patent " process for extracting of Olive leaf P.E and technology " (CN 1827127A) of the patent " a kind of Olive leaf P.E process for extracting " (CN 1583052A) of Yang Yinke application and Sun Haoyuan application; Two kinds of isolating product cleupins of patent institute are low, and all added exogenous material; The patent of Wang Chengzhang, high rosy clouds application " being rich in the preparation method of the leaf of Fructus oleae europaeae extraction of high purity cleupin " (CN 101003557A), the product of this patent preparation, content is higher than 50%, but far below 80%; Yao from the strange patent " technique for producing extractive of olive growing leaves " that waits application (CN101049328A), the isolating product of this patent, content are 2%～6%; Yao is from the strange patent " olive leaf extract and preparation method " (CN 101084945A) that waits application, though content the highest reach 95%, product yield is low, the production cycle is long, need leave standstill 24～36 hours; In addition; Adopt repeatedly organic solvent extraction, silica gel column chromatography and HPLC preparative hplc among the Wang Chengzhang etc. " introducing a fine variety Oleuropein extraction separation and structure evaluation in the A Si leaf of Fructus oleae europaeae "; Isolated content and be 95% cleupin, but this method is limited to the small amount of sample preparation to the greatest extent.Cleupin content is all lower in the isolating sample of U.S. Pat 5714150 and US98101563 institute, is no more than 40%.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a yield height is provided, the method for environmental protection a kind of extraction separation high-content cleupin from leaf of Fructus oleae europaeae.
The method of extraction separation high-content cleupin from leaf of Fructus oleae europaeae, concrete steps are following:
(1) with the leaf of Fructus oleae europaeae chopping, for cutting to 1～2cm;
(2) leaf of Fructus oleae europaeae with chopping adds purified water or aqueous ethanolic solution, extract at low temperature; The extract at low temperature temperature is 20～70 ℃ in said; Said extraction solvent is a purified water, and volume is 12～16 times; Said extraction solvent or be 40～80% ethanol, volume is 10～12 times;
(3) after the solution concentration after will extracting or directly through by two kinds of placed in-line chromatography columns of macroporous adsorbent resin; After the solution concentration after maybe will extracting or directly carry out primary sorption, carry out second adsorption after again desorbed solution being concentrated;
(4) with purified water resin is cleaned; Saidly with purified water resin is cleaned, volume is 8～20 times of resin column volume;
(5) with 5%～80% Fatty Alcohol(C12-C14 and C12-C18) wash-out resin, when effluent Fatty Alcohol(C12-C14 and C12-C18) concentration reaches 5%, begin to collect effluent; Said Fatty Alcohol(C12-C14 and C12-C18) is C
1-C
3Primary alconol and secondary alcohol, preferred alcohol, said Fatty Alcohol(C12-C14 and C12-C18) elution volume are 1-5 times of column volume;
(6) cryoconcentration elutriant; Said low temperature is 30-60 ℃, and said liquid concentrator concentration is 18～25%;
(7) the freezing or spraying drying of liquid concentrator; Can obtain cleupin.
Beneficial effect:
The compound yield is higher than prior art among the present invention, and the present invention compared with prior art it is advantageous that:
1. adopt the extract at low temperature technology, greatly reduce and extract required energy consumption, reduced the oxidation of cleupin, improved yield, also guaranteed the quality of product simultaneously;
2. adopt the separate mode of placed in-line column chromatography or secondary chromatography, need not to use noxious solvent (like ETHYLE ACETATE, chloroform, sherwood oil etc.), guaranteed the quality safety of product; Simplified technical process greatly; Simple to operation, production cost is low simultaneously, is beneficial to suitability for industrialized production;
3. the cleupin content of producing according to production technique provided by the invention detects through HPLC and can reach 80-90%, can satisfy the market requirement in fields such as modern functions food, healthcare products, has the good market advantage.
Embodiment
Be easy to understand and understand in order to make technique means of the present invention, creation characteristic, workflow, method of use reach purpose and effect,, further set forth the present invention below in conjunction with specific embodiment.
Embodiment 1
The leaf of Fructus oleae europaeae raw material is cut into 1～2cm; Place 14 times of purified water to its weight, 60 ℃ of extractions are filtered, filtrating with 0.8 times of column volume/hour flow velocity be the macroporous resin of HP-20 through model, absorption finishes, pure water is caught up with 5mins, closes 15mins.Clean resin with 10 times of purified water then to column volume; Again with resin LSA-8 or LSA-10 series connection; With 2 times to 30% ethanol of column volume with 1 times of column volume/hour flow velocity resolve; Treat that the effluent alcohol concn is to begin to collect desorbed solution at 5% o'clock; Being lower than 60 ℃, to be concentrated into concentration be 20%, and liquid concentrator adopts cryodesiccated method to carry out drying, obtains olive leaf extract.Detect cleupin content through HPLC and reach 85.26%.
Embodiment 2
The leaf of Fructus oleae europaeae raw material is cut into 1～2cm; Place 12 times of 40% ethanol to its weight, 50 ℃ of extractions are filtered, be lower than 60 ℃ and be concentrated into and do not have the alcohol flavor, liquid concentrator with 0.8 times of column volume/hour flow velocity be the macroporous resin of HP-20 through model, absorption finishes, pure water is caught up with 5mins, closes 15mins.Clean resin with 12 times of purified water then to column volume; Again with resin LSA-8 or LSA-10 series connection; With 3 times to 40% ethanol of column volume with 1 times of column volume/hour flow velocity resolve; Treat that the effluent alcohol concn is to begin to collect desorbed solution at 5% o'clock; Being lower than 60 ℃, to be concentrated into concentration be 18%, and liquid concentrator adopts spray-dired method to carry out drying, obtains olive leaf extract.Detect cleupin content through HPLC and reach 82.19%.
Embodiment 3
The leaf of Fructus oleae europaeae raw material is cut into 1～2cm; Place 10 times of 60% ethanol to its weight, 45 ℃ of extractions are filtered, be lower than 60 ℃ and be concentrated into and do not have the alcohol flavor, liquid concentrator with 0.8 times of column volume/hour flow velocity be the macroporous resin of HP-20 through model, absorption finishes, pure water is caught up with 5mins, closes 15mins.Clean resin with 10 times of purified water then to column volume; Again with 2.5 times to 25% ethanol of column volume with 1 times of column volume/hour flow velocity resolve, treat that the effluent alcohol concn is to begin to collect desorbed solution at 5% o'clock, be lower than 60 ℃ and be concentrated into and do not have the alcohol flavor; And then with 0.8 times of column volume/hour flow velocity be the macroporous resin of XAD-16 through model, absorption finishes, pure water is caught up with 5mins, closes 15mins.Clean resin with 10 times of purified water then to column volume; Again with 2 times to 60% ethanol of column volume with 1 times of column volume/hour flow velocity resolve; Treat that the effluent alcohol concn is to begin to collect desorbed solution at 5% o'clock; Desorbed solution is lower than 60 ℃, and to be concentrated into concentration be 22%, and liquid concentrator adopts cryodesiccated method to carry out drying, obtains olive leaf extract.Detect cleupin content through HPLC and reach 88.72%.
Embodiment 4
The leaf of Fructus oleae europaeae raw material is cut into 1～2cm; Place 15 times of purified water to its weight, 55 ℃ of extractions are filtered, filtrating with 0.8 times of column volume/hour flow velocity be the macroporous resin of XAD-16 through model, absorption finishes, pure water is caught up with 5mins, closes 15mins.Clean resin with 12 times of purified water then to column volume; Again with 3 times to 80% ethanol of column volume with 1 times of column volume/hour flow velocity resolve, treat that the effluent alcohol concn is to begin to collect desorbed solution at 5% o'clock, be lower than 60 ℃ and be concentrated into and do not have the alcohol flavor; And then with 0.8 times of column volume/hour flow velocity be the macroporous resin of HP-20 through model, absorption finishes, pure water is caught up with 5mins, closes 15mins.Clean resin with 10 times of purified water then to column volume; Again with 2 times to 60% ethanol of column volume with 1 times of column volume/hour flow velocity resolve; Treat that the effluent alcohol concn is to begin to collect desorbed solution at 5% o'clock; Being lower than 60 ℃, to be concentrated into concentration be 20%, and liquid concentrator adopts spray-dired method to carry out drying, obtains olive leaf extract.Detect cleupin content through HPLC and reach 84.60%.
More than show and described ultimate principle of the present invention and principal character and advantage of the present invention.The technician of the industry should understand; The present invention is not restricted to the described embodiments; That describes in the foregoing description and the specification sheets just explains principle of the present invention; Under the prerequisite that does not break away from spirit and scope of the invention, the present invention also has various changes and modifications, and these variations and improvement all fall in the scope of the invention that requires protection.The present invention requires protection domain to be defined by appending claims and equivalent thereof.
Claims (6)
1. the method for extraction separation high-content cleupin from leaf of Fructus oleae europaeae is characterized in that concrete steps are following:
(1) leaf of Fructus oleae europaeae is shredded;
(2) leaf of Fructus oleae europaeae with chopping adds purified water or aqueous ethanolic solution, extract at low temperature;
(3) after the solution concentration after will extracting or directly through by two kinds of placed in-line chromatography columns of macroporous adsorbent resin; After the solution concentration after maybe will extracting or directly carry out primary sorption, carry out second adsorption after again desorbed solution being concentrated;
(4) with purified water resin is cleaned;
(5) with 5%～80% Fatty Alcohol(C12-C14 and C12-C18) wash-out resin, when effluent Fatty Alcohol(C12-C14 and C12-C18) concentration reaches 5%, begin to collect effluent;
(6) cryoconcentration elutriant;
(7) the freezing or spraying drying of liquid concentrator.
2. according to claim 1 from leaf of Fructus oleae europaeae the method for extraction separation high-content cleupin, it is characterized in that: in the said step (1) with the leaf of Fructus oleae europaeae chopping, for cutting to 1～2cm.
3. according to claim 1 from leaf of Fructus oleae europaeae the method for extraction separation high-content cleupin, it is characterized in that: the extract at low temperature temperature is 20～70 ℃ in the said step (2); Said extraction solvent is a purified water, and volume is 12～16 times; Said extraction solvent or be 40～80% ethanol, volume is 10～12 times.
4. according to claim 1 from leaf of Fructus oleae europaeae the method for extraction separation high-content cleupin, it is characterized in that: said step is cleaned resin with purified water in (4), volume is 8～20 times of resin column volume.
5. according to claim 1 from leaf of Fructus oleae europaeae the method for extraction separation high-content cleupin, it is characterized in that: Fatty Alcohol(C12-C14 and C12-C18) is C1-C3 primary alconol and secondary alcohol in the said step (5), and preferred alcohol, said Fatty Alcohol(C12-C14 and C12-C18) elution volume are 1-5 times of column volume.
6. according to claim 1 from leaf of Fructus oleae europaeae the method for extraction separation high-content cleupin, it is characterized in that: low temperature is 30-60 ℃ in the said step (6), and said liquid concentrator concentration is 18～25%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110439186.7A CN102532217B (en) | 2011-12-23 | 2011-12-23 | Method for purifying and separating high-content oleuropein from olive leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110439186.7A CN102532217B (en) | 2011-12-23 | 2011-12-23 | Method for purifying and separating high-content oleuropein from olive leaves |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102532217A true CN102532217A (en) | 2012-07-04 |
| CN102532217B CN102532217B (en) | 2014-12-10 |
Family
ID=46340374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110439186.7A Active CN102532217B (en) | 2011-12-23 | 2011-12-23 | Method for purifying and separating high-content oleuropein from olive leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102532217B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333213A (en) * | 2013-07-24 | 2013-10-02 | 桂林茗兴生物科技有限公司 | Method for extracting oleuropein from olive leaves |
| CN104311613A (en) * | 2014-09-30 | 2015-01-28 | 桂林三宝药业有限公司 | Method for extracting oleuropein from olive leaves |
| CN105011162A (en) * | 2015-07-02 | 2015-11-04 | 罗新华 | Olive leaf essence as well as making technology and application thereof |
| CN106565803A (en) * | 2016-10-10 | 2017-04-19 | 广西来宾绿翔生物科技有限公司 | Extraction process for oleuropein |
| CN109758498A (en) * | 2019-03-21 | 2019-05-17 | 湖南朗林生物资源股份有限公司 | A kind of preparation method of extractive of olive growing leaves |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002128678A (en) * | 2000-10-17 | 2002-05-09 | Tama Seikagaku Kk | Method for producing extract composition containing oleuropein |
| WO2008136037A2 (en) * | 2007-05-04 | 2008-11-13 | Università Degli Studi Magna Graecia Di Catanzaro | Chemical-catalytic method for the peracylation of oleuropein and its products of hydrolysis |
| CN101781344A (en) * | 2009-12-13 | 2010-07-21 | 成都普思生物科技有限公司 | Method for preparing olive leaf extract |
| CN101955503A (en) * | 2010-05-25 | 2011-01-26 | 苏州派腾生物医药科技有限公司 | Preparation method of oleuropein |
| CN102060893A (en) * | 2009-11-13 | 2011-05-18 | 中国科学院兰州化学物理研究所 | Process method for synchronously preparing oleuropein and olive total flavonoids from olive leaves |
-
2011
- 2011-12-23 CN CN201110439186.7A patent/CN102532217B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002128678A (en) * | 2000-10-17 | 2002-05-09 | Tama Seikagaku Kk | Method for producing extract composition containing oleuropein |
| WO2008136037A2 (en) * | 2007-05-04 | 2008-11-13 | Università Degli Studi Magna Graecia Di Catanzaro | Chemical-catalytic method for the peracylation of oleuropein and its products of hydrolysis |
| CN102060893A (en) * | 2009-11-13 | 2011-05-18 | 中国科学院兰州化学物理研究所 | Process method for synchronously preparing oleuropein and olive total flavonoids from olive leaves |
| CN101781344A (en) * | 2009-12-13 | 2010-07-21 | 成都普思生物科技有限公司 | Method for preparing olive leaf extract |
| CN101955503A (en) * | 2010-05-25 | 2011-01-26 | 苏州派腾生物医药科技有限公司 | Preparation method of oleuropein |
Non-Patent Citations (1)
| Title |
|---|
| 党建章，等: "D-101大孔树脂富集橄榄叶中橄榄苦苷的研究", 《中药材》, vol. 30, no. 4, 30 April 2007 (2007-04-30), pages 454 - 457 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333213A (en) * | 2013-07-24 | 2013-10-02 | 桂林茗兴生物科技有限公司 | Method for extracting oleuropein from olive leaves |
| CN103333213B (en) * | 2013-07-24 | 2015-10-07 | 桂林茗兴生物科技有限公司 | The method of oleuropein is extracted from leaf of Fructus oleae europaeae |
| CN104311613A (en) * | 2014-09-30 | 2015-01-28 | 桂林三宝药业有限公司 | Method for extracting oleuropein from olive leaves |
| CN105011162A (en) * | 2015-07-02 | 2015-11-04 | 罗新华 | Olive leaf essence as well as making technology and application thereof |
| CN106565803A (en) * | 2016-10-10 | 2017-04-19 | 广西来宾绿翔生物科技有限公司 | Extraction process for oleuropein |
| CN109758498A (en) * | 2019-03-21 | 2019-05-17 | 湖南朗林生物资源股份有限公司 | A kind of preparation method of extractive of olive growing leaves |
| CN109758498B (en) * | 2019-03-21 | 2021-08-17 | 湖南朗林生物资源股份有限公司 | Preparation method of olive leaf extract |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102532217B (en) | 2014-12-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102250195B (en) | Method for producing xanthoceraside | |
| CN101805410B (en) | Technological method for comprehensively producing pueraria flavonid, puerarin and arrowroot starch | |
| CN105055527A (en) | Preparation method of olive leaf extract | |
| CN102532217B (en) | Method for purifying and separating high-content oleuropein from olive leaves | |
| CN102451235A (en) | Preparation method of olive leaf extract | |
| CN110818668B (en) | Method for separating and purifying high-purity flavonoid glycoside compounds from gynura procumbens stems | |
| CN105326876A (en) | Method for extracting total flavonoids of chrysanthemum | |
| CN104523836A (en) | Method for comprehensively extracting protein, polysaccharide and polyphenol from avocado kernels | |
| CN107759648B (en) | Method for separating and purifying hyperoside and isoquercitrin from abelmoschus manihot | |
| CN107325138A (en) | A kind of method of the main anthocyanin of four kinds of extraction separation and purification in pomace from blackcurrant | |
| CN102659864B (en) | Preparation method of anti-allergic natural product of 2-O-alpha-L-rhamnose-4,6,4'-trihydroxybenzophenone (THBP) | |
| CN114515008A (en) | Cistanche tubulosa extract and preparation method thereof | |
| CN102627679A (en) | Method for preparing schaftoside from desmodium styracifolium | |
| CN102731592A (en) | Method for extracting cleupin and amentoflavone from olive leaf | |
| CN105175426B (en) | A kind of method of the extraction purification Bergenin from treebine stem | |
| CN1872870A (en) | Method for preparing oleanolic acid from wrinkled papaya | |
| CN101530193A (en) | Method for comprehensively extracting and purifying oxidation-resistant active ingredient in sugarcane toppers or slag | |
| CN102274386A (en) | Method for extracting total isoflavones from Allium mongolicum Regel | |
| CN111892503B (en) | Method for rapidly preparing high-purity chlorogenic acid from gynura procumbens | |
| CN106986904A (en) | A kind of Gastrodin extracted in the anesthesia stem from day and preparation method thereof | |
| CN104586917A (en) | Method for extracting total flavonoid from dragon fruit peels | |
| CN103804193A (en) | Method for extracting chlorogenic acid and pinoresinol from eucommia barks | |
| CN102648939A (en) | Preparation method for geum aleppicum polyphenol | |
| CN101450962B (en) | Method for extracting oleanolic acid from Kandelia candel leaf | |
| CN106256353A (en) | A kind of method of isolated and purified chlorogenic acid in mulberry leaves constituents |
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 | ||
| DD01 | Delivery of document by public notice | ||
| DD01 | Delivery of document by public notice |
Addressee: Wang Keming Document name: Notification of Decision on Request for Restoration of Right |
|
| DD01 | Delivery of document by public notice | ||
| DD01 | Delivery of document by public notice |
Addressee: Wang Keming Document name: Notification to Pay the Fees |