CN101906059B - Method for refining astaxanthin from crude extracts containing ester astaxanthin - Google Patents
Method for refining astaxanthin from crude extracts containing ester astaxanthin Download PDFInfo
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- CN101906059B CN101906059B CN201010235331A CN201010235331A CN101906059B CN 101906059 B CN101906059 B CN 101906059B CN 201010235331 A CN201010235331 A CN 201010235331A CN 201010235331 A CN201010235331 A CN 201010235331A CN 101906059 B CN101906059 B CN 101906059B
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- astaxanthin
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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 method that contains refining astaxanthin in the ester astaxanthin crude extract that particularly 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 carrotenoid, and its chemical name is 3,3 ,-dihydroxyl-β, and β ,-Serlabo-4,4 ,-diketone, molecular formula is C
40H
520
4, relative molecular mass is 596.86.Astaxanthin has very strong oxidation-resistance and the ability of removing 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 main with chemosynthesis, and its technology is very ripe.But the progressively raising of Along with people's standard of living and environmental consciousness has become the research focus to from the organism of astaxanthin-containing, extracting astaxanthin.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 from the organism of astaxanthin-containing, extracting astaxanthin mainly concentrates on slightly puies forward the aspect, like 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; From Haematocoocus Pluvialls, adopt supercritical extraction etc. in the patent 200910102291.4, 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 from organism, extract 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% inhibitor, volume ratio and be the methanol solution or the sodium hydroxide ethanolic soln of the Pottasium 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 through 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: with the mixed solution of saponification liquor and washings be adsorbed onto nonpolar with the flow velocity of 0.5～2BV/h or the low-pole macroporous adsorbent resin on; With 2BV zero(ppm) 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 waters by volume, carry out crystallization at vacuum state, 30～60 ℃ the time; To the solid-liquid volume ratio is to stop in 1～50: 100 o'clock, when controlled temperature is reduced to-10～4 ℃, educates 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.
Inhibitor is one or more in butyl hydroxy anisol (BHA), 2,6 di tert butyl 4 methyl phenol (BHT), S-WAT, sodium sulfite anhy 96, Sodium Pyrosulfite, the Sulfothiorine in the said saponification step.
Eluent is one or more of acetone, chloroform, ethyl acetate solution in the said resin purification step.
Resin in the said 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 be prone to 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 get into 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, through 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 inhibitor 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; Step is simple and be prone to amplification, 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 embodiment.
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.12No.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 metals ion; Add 0.002% butyl hydroxy anisol (BHA) then, according to the ethanolic soln of volume ratio adding in 1: 1 10g/L sodium hydroxide, 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 through 0.45 μ m membrane filtration;
Adopt the nonpolar macroporous adsorption resin HPDI00 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 zero(ppm) 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, educate in the brilliant groove controlled temperature at temperature control again and educated brilliant 8 hours under for-10 ℃ of conditions, 3000g is centrifugal then throws away to absence of liq; Use deionized water wash twice again, collect 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 metals ion, add 0.01% 2,6 di tert butyl 4 methyl phenol (BHT) then; 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 through 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 zero(ppm) 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, educate at temperature control that controlled temperature is to educate under 0 ℃ of condition brilliant 12 hours in the brilliant groove again, 5000g is centrifugal then throws away to absence of liq; Use deionized water wash twice again, collect 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 from crustaceans (like the shrimp crab) processing waste water, to adopt flocculence at 50kg; Jiang Guoliang, Li Lide, Yang Dong, Liu Yun; Number of patent application: in the crude extract that contains ester astaxanthin that 02135919.9) obtains, added 0.02% EDTA earlier in order to chelating Fe
3+, Cu
2+Deng metals ion, add the 2,6 di tert butyl 4 methyl phenol (BHT) of 0.01% butyl hydroxy anisol (BHA) and 0.02% then; The methanol solution that adds 10g/L Pottasium 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 through 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 zero(ppm) water wash-out; With KOH, methyl alcohol and some other impurity flush awaies that can not adsorb; Use ETHYLE 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, educate at temperature control that controlled temperature is to educate under 4 ℃ of conditions brilliant 24 hours in the brilliant groove again, 7000g is centrifugal then throws away to absence of liq; Use deionized water wash twice again, collect 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% inhibitor, 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, and added 2～20 times of corresponding methyl alcohol of volume or ethanol after the saponification again, and remove impurity through 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: with the mixed solution of saponification liquor and washings be adsorbed onto nonpolar with the flow velocity of 0.5～2BV/h or the low-pole macroporous adsorbent resin on; With 2BV zero(ppm) 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, add 10～50% de-salted water or deionized water, carry out crystallization at vacuum state, 30～60 ℃ the time; To the solid-liquid volume ratio is to stop in 1～50: 100 o'clock, when controlled temperature is reduced to-10～4 ℃, educates 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 that contains ester astaxanthin the method for refining astaxanthin; It is characterized in that: inhibitor is one or more in butyl hydroxy anisol (BHA), 2,6 di tert butyl 4 methyl phenol (BHT), S-WAT, sodium sulfite anhy 96, Sodium Pyrosulfite, the Sulfothiorine in the said saponification step.
3. according to claim 1 from the crude extract that contains ester astaxanthin the method for refining astaxanthin, it is characterized in that: eluent is one or more of acetone, chloroform, ethyl acetate solution in the said resin purification step.
4. according to claim 1 from the crude extract that contains ester astaxanthin the method for refining astaxanthin, it is characterized in that: the resin in the said resin purification step is used Ethanol Treatment in advance.
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| CN102659652B (en) * | 2012-04-12 | 2014-08-06 | 西北农林科技大学 | Solid phase extraction method for extracting total astaxanthin from haematococcus pluvialis |
| CN103044304B (en) * | 2012-12-21 | 2014-12-03 | 宁波红龙生物科技有限公司 | Method for preparing astaxanthin extractive from haematococcus pluvialis powder |
| CN107602434B (en) * | 2017-09-21 | 2020-02-21 | 嘉必优生物技术（武汉）股份有限公司 | Astaxanthin crystal derived from haematococcus pluvialis and preparation process thereof |
| CN108588161B (en) * | 2018-04-16 | 2021-08-20 | 昆明理工大学 | New application of 2, 6-di-tert-butyl-4-methylphenol |
| 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 (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1548420A (en) * | 2003-05-17 | 2004-11-24 | 浙江工业大学 | Extraction process of astaxanthin in phaffiarhodozyma |
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| CN1548420A (en) * | 2003-05-17 | 2004-11-24 | 浙江工业大学 | Extraction process of astaxanthin in phaffiarhodozyma |
Non-Patent Citations (3)
| Title |
|---|
| JP特开2010-43052A 2010.02.25 |
| 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)》.2005,第33卷(第2期),264－268. |
| 雨生红球藻中虾青素酯的皂化及游离虾青素的纯化分离;陈兴才等;《福州大学学报(自然科学版)》;20050430;第33卷(第2期);264－268 * |
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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 |
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