CN101906059A - Method for refining astaxanthin from crude extracts containing ester astaxanthin - Google Patents
Method for refining astaxanthin from crude extracts containing ester astaxanthin Download PDFInfo
- Publication number
- CN101906059A CN101906059A CN2010102353315A CN201010235331A CN101906059A CN 101906059 A CN101906059 A CN 101906059A CN 2010102353315 A CN2010102353315 A CN 2010102353315A CN 201010235331 A CN201010235331 A CN 201010235331A CN 101906059 A CN101906059 A CN 101906059A
- Authority
- CN
- China
- Prior art keywords
- astaxanthin
- refining
- saponification
- crude extract
- deionized water
- 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
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a method for refining astaxanthin from crude extracts containing ester astaxanthin. The method can be used for obtaining an astaxanthin finished product with the purity above 90% by adopting the steps of saponification, resin purification, crystallization refining, and the like, and the yield of the finished product is above 60%. The method has the characteristics of simple operation steps, large-scale industrial production, environmental protection, and the like, and the obtained astaxanthin product has higher purity and yield.
Description
Technical field
The present invention relates to a kind of from crude extracts containing ester astaxanthin the method for refining astaxanthin, the particularly method that contains refining astaxanthin in the ester astaxanthin crude extract that from the unit cell algae, yeast and the fishery products waste that contain astaxanthin, obtains through methods such as organic solvent extraction and flocculations.
Background technology
Astaxanthin (Astaxanthin) belongs to the terpenes unsaturated compound, is a kind of keto-acid carotenoid, and its chemical name is 3,3 ,-dihydroxyl-β, and β ,-carotene-4, the 4.-diketone, molecular formula is C
40H
52O
4, relative molecular mass is 596.86.Astaxanthin has very strong oxidation-resistance and removes the ability of free radical, its health to human body plays an important role, can prevent the oxidized damage of tissue, cell and DNA effectively, therefore be widely used in industries such as food, medicine, makeup and healthcare products.
The industrial production astaxanthin is based on chemosynthesis, and its technology is very ripe.But, become the research focus to from the organism of astaxanthin-containing, extracting astaxanthin along with the progressively raising of people's living standard and environmental consciousness.And in the high biology of content astaxanthin, like rain giving birth to haematococcus pulvialis, phaffiafhodozyma etc., the astaxanthin great majority are to exist with ester class form, like rain the astaxanthin of giving birth to the dibasic acid esters form that just contains 70% monoesters and 25% in the haematococcus pulvialis.
The at present domestic technology of extracting astaxanthin from the organism of astaxanthin-containing mainly concentrates on slightly puies forward the aspect, as the employing flocculence extraction method of introducing in the patent 02135919.9 from chitin factory process waste water; From fresh shrimp shell and huge legendary turtle shrimp shell, adopt acid to add organic solvent extration among patent 02122565.6 and the 200810236028.X; Adopt supercritical extraction etc. in the patent 200910102291.4 from Haematocoocus Pluvialls, it is lower that these methods make finished product astaxanthin purity, the highlyest can only reach 60%.In addition, the Liu Li of Northeast Forestry University Na (Northeast Forestry University's master thesis, 2006) though make product purity obtain the raising of certain amplitude through steps such as silica gel column chromatography and crystallizations, silica gel column chromatography technology is difficult for scale amplifies, the difficult industrialization demands that adapts to.
Because the restriction of Technology, the astaxanthin commodity that extract from organism of domestic sale mostly are effective contents at 1～10% thick finished product.Therefore but developing a kind of technology of extracting high purity astaxanthin suitability for industrialized production has boundless prospect.
Summary of the invention
In view of this, the purpose of this invention is to provide a kind of from crude extracts containing ester astaxanthin the method for refining astaxanthin, this method have operation steps simple, can large-scale industrial production, characteristics such as environmental protection.
The present invention adopts following technical scheme: adopt crude extracts containing ester astaxanthin as raw material, carry out the operation of following steps:
Saponification: contain in the crude extract of ester astaxanthin at 2～100kg, add 0.004～0.04% EDTA, 0.002～0.1% antioxidant, volume ratio and be the methanol solution or the sodium hydroxide ethanolic soln of the potassium hydroxide of 1～4: 2 10g/L, controlled temperature is 0～60 ℃, stirred 0.5～12 hour, add 2～10 times of corresponding methyl alcohol of volume or ethanol after the saponification again and remove impurity by 0.45 μ m membrane filtration, and with corresponding methyl alcohol or washing with alcohol filter membrane impurity till color is turned white, merge saponification liquor and washings;
Resin purification: be adsorbed onto nonpolar with the flow velocity of 0.5～2BV/h the mixed solution of saponification liquor and washings or the low-pole macroporous adsorbent resin on, with 2BV distilled water or deionized water wash-out, use the eluent wash-out again, the control flow velocity is 0.2～1BV/h, collects the red solution of astaxanthin-containing;
Crystal refining: in the astaxanthin-containing red solution of collecting, 10～50% add de-salted water or deionized water by volume, carrying out crystallization at vacuum state, 30～60 ℃ the time, is to stop in 1～50: 100 o'clock to the solid-liquid volume ratio, when controlled temperature is reduced to-10～4 ℃, educated brilliant 2～24 hours, 3000～7000g is centrifugal then, uses the deionized water wash secondary again, collects crystal, vacuum-drying 4～12 hours in the time of 30～60 ℃ again obtains the finished product.
Antioxidant is a butyl hydroxy anisol (BHA), 2 in the described saponification step, one or more in 6-di-t-butyl hydroxytoluene (BHT), S-WAT, sodium bisulfite, Sodium Pyrosulfite, the Sulfothiorine.
Eluent is one or more of acetone, chloroform, ethyl acetate solution in the described resin purification step.
Resin in the described resin purification step is used Ethanol Treatment in advance, because macroporous resin adsorption is the material of molecular state, molecular state impurity is transferred to be dissolved in ethanol, thereby easily be taken away by ethanol eluate, recovers the adsorptive capacity of resin.
In addition, the mother liquor that centrifugal process produces in the last crystal refining step can enter the upper prop absorption again of resin purification step once more; Do not dissolve in the resin purification wash-out when requiring and carry out secondary recrystallization in solvent when purity reaches, thereby further improve product purity, the waste liquid that reduces contaminate environment produces.
The present invention has the following advantages:
(1) contain in the organism of ester astaxanthin, astaxanthin with the existing of monoesters and dibasic acid esters form, by saponification step, makes these astaxanthin esters become the astaxanthin of free shape mostly, and follow-up being further purified laid the foundation; (2) saponification step has added a certain amount of EDTA among the present invention, with the Fe in the astaxanthin crude extract
3+, Cu
2+Deng metal ion-chelant, can slow down the speed of astaxanthin oxidation; Also add a certain amount of antioxidant in addition, prevented the oxidation of astaxanthin effectively; (3) operations such as saponification, absorption, crystallization are combined effectively, some unnecessary steps such as silica gel column chromatography etc. have been removed, simple and the easily amplification of step has significantly improved astaxanthin product yield and purity, and the batch processing amount also meets industrial production requirement; (4) solve the handling problem of crystalline mother solution effectively, improved product yield.
In sum, the present invention have operation steps simple, can large-scale industrial production, characteristics such as environmental protection, and final astaxanthin purity and yield are higher.
Embodiment
Describe in conjunction with specific embodiments.
Embodiment 1
2kg adopt from Haematocoocus Pluvialls ethyl acetate-ethanol (1: 2) method (method with reference to Haematocoocus Pluvialls the research of astaxanthin ester saponification process, Zhou Shuanlin, Zhang Lifei, the China pharmacist, Vol.12N0.8 in 2009,1018～1020) obtain and in the spissated crude extract that contains ester astaxanthin, add 0.004% EDTA earlier in order to chelating Fe
3+, Cu
2+Deng metal ion, add 0.002% butyl hydroxy anisol (BHA) then, the ethanolic soln that adds 10g/L sodium hydroxide according to volume ratio at 1: 1, the control saponification temperature is 20 ℃, fill under the nitrogen condition in lucifuge, sealing and to stir 3 hours, add 3 times of corresponding methyl alcohol of volume after the saponification again and remove impurity, and wash filter membrane impurity till color is turned white, merge saponification liquor and washings with corresponding methyl alcohol by 0.45 μ m membrane filtration;
Adopt the nonpolar macroporous adsorption resin HPD100 type dress post of crossing through ethanol elution, use deionized water flush away ethanol then, the saponification liquor that obtains in the last step and the mixed solution of washings are adsorbed onto on the resin with the flow velocity of 0.5BV/h, with 2BV distilled water wash-out, with KOH, methyl alcohol and some other impurity flush awaies that can not adsorb, use the acetone wash-out again, the control flow velocity is 0.2BV/h, obtain solution and be divided into green and red two portions, collect red solution.
In the red liquid of collecting, 20% add de-salted water by volume, in vacuum cyrsytalliser, be controlled under the vacuum condition, carry out crystallization when temperature is 30 ℃, stopped to concentrate in 1: 40 o'clock to the solid-liquid volume ratio, educating in the brilliant groove controlled temperature at temperature control again educated brilliant 8 hours under for-10 ℃ of conditions, 3000g is centrifugal then throws away to absence of liquid, uses deionized water wash twice again, collects crystal, in the time of 30 ℃, carry out vacuum-drying, obtain final finished, wherein content astaxanthin 90%, and yield reaches 70%.
Embodiment 2
30kg from phaffiafhodozyma through 80% ethanol repeatedly extract (method is with reference to Northeast Forestry University, Liu Lina, master thesis, 2006) concentrate after gained contain the crude extract of ester astaxanthin, added 0.01% EDTA earlier in order to chelating Fe
3+, Cu
2+Deng metal ion, add then 0.01% 2,6-di-t-butyl hydroxytoluene (BH T), the ethanolic soln that adds 10g/L sodium hydroxide according to volume ratio at 1: 2, the control saponification temperature is 30 ℃, fills under the nitrogen condition in lucifuge, sealing and stirs 4 hours, adds 4 times of corresponding ethanol of volume after the saponification again and removes impurity by 0.45 μ m membrane filtration, and with washing with alcohol filter membrane impurity till color is turned white, merge saponification liquor and washings;
Adopt the nonpolar macroporous adsorption resin dress post of crossing through ethanol elution, use deionized water flush away ethanol then, the saponification liquor that obtains in the last step and the mixed solution of washings are adsorbed onto on the resin with the flow velocity of 0.5BV/h, with 2BV distilled water wash-out, with sodium hydroxide, ethanol and some other impurity flush awaies that can not adsorb, use the chloroform wash-out again, the control flow velocity is 0.5BV/h, collects red solution.
In the red liquid of collecting, 30% add de-salted water by volume, in vacuum cyrsytalliser, be controlled under the vacuum condition, carry out crystallization when temperature is 50 ℃, stopped to concentrate in 1: 50 o'clock to the solid-liquid volume ratio, educating in the brilliant groove controlled temperature at temperature control again is to educate under 0 ℃ of condition brilliant 12 hours, 5000g is centrifugal then throws away to absence of liquid, uses deionized water wash twice again, collects crystal, in the time of 50 ℃, carry out vacuum-drying, obtain final finished, wherein content astaxanthin 93%, and yield reaches 68%.
Embodiment 3
(method is with reference to a kind of production technique of astaxanthin to adopt flocculence at 50kg from crustaceans (as the shrimp crab) processing waste water, Jiang Guoliang, Li Lide, Yang Dong, Liu Yun, number of patent application: 02135919.9) in the crude extract that contains ester astaxanthin of Huo Deing, added 0.02% EDTA earlier in order to chelating Fe
3+, Cu
2+Deng metal ion, add 2 of 0.01% butyl hydroxy anisol (BHA) and 0.02% then, 6-di-t-butyl hydroxytoluene (BHT), the methanol solution that adds 10g/L potassium hydroxide according to volume ratio at 1: 3, the control saponification temperature is 60 ℃, fills under the nitrogen condition in lucifuge, sealing and stirs 3 hours, adds 5 times of corresponding methyl alcohol of volume after the saponification again and removes impurity by 0.45 μ m membrane filtration, and with methanol wash filter membrane impurity till color is turned white, merge saponification liquor and washings;
Adopt the low-pole macroporous adsorbent resin dress post of crossing through ethanol elution, use deionized water flush away ethanol then, the saponification liquor that obtains in the last step and the mixed solution of washings are adsorbed onto on the resin with the flow velocity of 2BV/h, with 2BV distilled water wash-out, with KOH, methyl alcohol and some other impurity flush awaies that can not adsorb, use ethyl acetate again: acetone (3: 1) wash-out, the control flow velocity is 1BV/h, collects red solution.
In the red liquid of collecting, 50% add deionized water by volume, in vacuum cyrsytalliser, be controlled under the vacuum condition, carry out crystallization when temperature is 60 ℃, stopped to concentrate in 1: 60 o'clock to the solid-liquid volume ratio, educating in the brilliant groove controlled temperature at temperature control again is to educate under 4 ℃ of conditions brilliant 24 hours, 7000g is centrifugal then throws away to absence of liquid, uses deionized water wash twice again, collects crystal, in the time of 60 ℃, carry out vacuum-drying, obtain final finished, wherein content astaxanthin 95%, and yield reaches 65%.
Claims (4)
1. the method for a refining astaxanthin from crude extracts containing ester astaxanthin is characterized in that: adopt the crude extract that contains ester astaxanthin as raw material, carry out the operation of following steps:
Saponification: contain in the crude extract of ester astaxanthin at 2～100kg, add 0.004～0.04% EDTA, 0.002～0.02% antioxidant, volume ratio and be potassium hydroxide methanol solution or the sodium hydroxide ethanolic soln of 1～4: 2 10g/L, controlled temperature is 0～60 ℃, stirred 0.5～12 hour, add 2～20 times of corresponding methyl alcohol of volume or ethanol after the saponification again, and remove impurity by 0.45 μ m membrane filtration, and till turning white to color with corresponding methyl alcohol or washing with alcohol filter membrane impurity, merge saponification liquor and washings;
Resin purification: be adsorbed onto nonpolar with the flow velocity of 0.5～2BV/h the mixed solution of saponification liquor and washings or the low-pole macroporous adsorbent resin on, with 2BV distilled water or deionized water wash-out, use the eluent wash-out again, the control flow velocity is 0.2～1BV/h, collects the red solution of astaxanthin-containing;
Crystal refining: in the astaxanthin-containing red solution of collecting, the de-salted water or the deionized water of adding 10～50%, carrying out crystallization at vacuum state, 30～60 ℃ the time, is to stop in 1～50: 100 o'clock to the solid-liquid volume ratio, when controlled temperature is reduced to-10～4 ℃, educated brilliant 2～24 hours, 3000～7000g is centrifugal then, uses the deionized water wash secondary then, collects crystal, vacuum-drying 4～12 hours in the time of 30～60 ℃ again obtains the finished product.
2. according to claim 1 from the crude extract of astaxanthin-containing the method for refining astaxanthin, it is characterized in that: antioxidant is a butyl hydroxy anisol (BHA), 2 in the described saponification step, one or more in 6-di-t-butyl hydroxytoluene (BHT), S-WAT, sodium bisulfite, Sodium Pyrosulfite, the Sulfothiorine.
3. according to claim 1 from the crude extract of astaxanthin-containing the method for refining astaxanthin, it is characterized in that: eluent is one or more of acetone, chloroform, ethyl acetate solution in the described resin purification step.
4. according to claim 1 from the crude extract of astaxanthin-containing the method for refining astaxanthin, it is characterized in that: the resin in the described resin purification step is used Ethanol Treatment in advance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010235331A CN101906059B (en) | 2010-07-23 | 2010-07-23 | Method for refining astaxanthin from crude extracts containing ester astaxanthin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010235331A CN101906059B (en) | 2010-07-23 | 2010-07-23 | Method for refining astaxanthin from crude extracts containing ester astaxanthin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101906059A true CN101906059A (en) | 2010-12-08 |
| CN101906059B CN101906059B (en) | 2012-10-17 |
Family
ID=43261671
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010235331A Expired - Fee Related CN101906059B (en) | 2010-07-23 | 2010-07-23 | Method for refining astaxanthin from crude extracts containing ester astaxanthin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101906059B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659652A (en) * | 2012-04-12 | 2012-09-12 | 西北农林科技大学 | Solid phase extraction method for extracting total astaxanthin from haematococcus pluvialis |
| CN103044304A (en) * | 2012-12-21 | 2013-04-17 | 宁波红龙生物科技有限公司 | Method for preparing astaxanthin extractive from haematococcus pluvialis powder |
| CN107602434A (en) * | 2017-09-21 | 2018-01-19 | 嘉必优生物技术（武汉）股份有限公司 | The astaxanthin crystal and its preparation technology in haematococcus pluvialis source |
| CN108588161A (en) * | 2018-04-16 | 2018-09-28 | 昆明理工大学 | The new application of 2,6 di tert butyl 4 methyl phenol |
| CN110522666A (en) * | 2019-09-26 | 2019-12-03 | 杭州兰茜化妆品有限公司 | A kind of astaxanthin Essence and its preparation process |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1548420A (en) * | 2003-05-17 | 2004-11-24 | 浙江工业大学 | Extraction process of astaxanthin in phaffiarhodozyma |
| JP2010043052A (en) * | 2008-08-18 | 2010-02-25 | Mitsubishi Chemicals Corp | Separation method of carotenoids |
-
2010
- 2010-07-23 CN CN201010235331A patent/CN101906059B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1548420A (en) * | 2003-05-17 | 2004-11-24 | 浙江工业大学 | Extraction process of astaxanthin in phaffiarhodozyma |
| JP2010043052A (en) * | 2008-08-18 | 2010-02-25 | Mitsubishi Chemicals Corp | Separation method of carotenoids |
Non-Patent Citations (1)
| Title |
|---|
| 陈兴才等: "雨生红球藻中虾青素酯的皂化及游离虾青素的纯化分离", 《福州大学学报(自然科学版)》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659652A (en) * | 2012-04-12 | 2012-09-12 | 西北农林科技大学 | Solid phase extraction method for extracting total astaxanthin from haematococcus pluvialis |
| CN103044304A (en) * | 2012-12-21 | 2013-04-17 | 宁波红龙生物科技有限公司 | Method for preparing astaxanthin extractive from haematococcus pluvialis powder |
| CN107602434A (en) * | 2017-09-21 | 2018-01-19 | 嘉必优生物技术（武汉）股份有限公司 | The astaxanthin crystal and its preparation technology in haematococcus pluvialis source |
| CN108588161A (en) * | 2018-04-16 | 2018-09-28 | 昆明理工大学 | The new application of 2,6 di tert butyl 4 methyl phenol |
| CN110522666A (en) * | 2019-09-26 | 2019-12-03 | 杭州兰茜化妆品有限公司 | A kind of astaxanthin Essence and its preparation process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101906059B (en) | 2012-10-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101906059B (en) | Method for refining astaxanthin from crude extracts containing ester astaxanthin | |
| CN102775523B (en) | Process for preparing high-purity low-molecular heparin sodium | |
| CN1321961C (en) | New method for picking-up purified resveratrol from giant knotweed | |
| CN110790805B (en) | Method for extracting chenodeoxycholic acid from pig bile paste | |
| CN101613390A (en) | A kind of method of separating and purifying high-purity cordycepin | |
| CN102516325A (en) | Method for producing crocin with higher than 95% purity from gardenia | |
| CN1869043A (en) | Synthesis method of chenodeoxycholic acid | |
| CN102267906B (en) | Extraction method for chlorogenic acid | |
| CN106755214B (en) | Method for obtaining baohuoside I by means of two-phase enzymatic hydrolysis | |
| CN104628802B (en) | Method for extracting and purifying nemadectin from fermentation liquid | |
| CN104119229A (en) | Technology for producing pure chlorogenic acid | |
| CN106561722A (en) | Algal inhibition application of undaria pinnatifida and separation and purification method of undaria pinnatifida algal inhibition active substance | |
| CN102002072A (en) | Process for extracting flavone from date pit | |
| CN110396058A (en) | A kind of novel calcifediol (25-hydroxyvitamin D3) isolation and purification method | |
| CN113354526B (en) | Alkali purification method of coenzyme Q10 | |
| CN101434522B (en) | Method for preparing high-purity gossypol from cottonseed dephenolizing solution | |
| CN101353294A (en) | Separation and purification method of high-content resveratrol | |
| CN104418825A (en) | Method for purifying lipstatin | |
| CN113912480A (en) | Extraction method of coenzyme Q10 | |
| CN111943827A (en) | Method for purifying coenzyme Q10 | |
| CN103130916A (en) | Novel technology for extracting high purity heparin sodium from intestinal mucosa | |
| CN115286678B (en) | Purification method of bioconversion tauroursodeoxycholic acid | |
| CN105175471A (en) | Method for extracting androstenedione crude product from two-liquid-phase system | |
| TWI568366B (en) | Extracting method | |
| CN103305572B (en) | Method for producing baohuoside I through herba epimdii |
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 | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Method for refining astaxanthin from crude extracts containing ester astaxanthin Effective date of registration: 20131128 Granted publication date: 20121017 Pledgee: Pudong Shanghai Development Bank Limited by Share Ltd Tianjin branch Pledgor: Tianjin Qiangweite Bio-Tech Co., Ltd. Registration number: 2013120000054 |
|
| PLDC | Enforcement, change and cancellation of contracts on pledge of patent right or utility model | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20121017 Termination date: 20150723 |
|
| EXPY | Termination of patent right or utility model |