CN102731358B - Method for extracting astaxanthin diester from antarctic krill - Google Patents
Method for extracting astaxanthin diester from antarctic krill Download PDFInfo
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- CN102731358B CN102731358B CN201210248678.2A CN201210248678A CN102731358B CN 102731358 B CN102731358 B CN 102731358B CN 201210248678 A CN201210248678 A CN 201210248678A CN 102731358 B CN102731358 B CN 102731358B
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Abstract
The invention discloses a method for extracting astaxanthin diester from antarctic krill, which comprises the following steps of: adding n-hexane into dried antarctic krill, stirring, extracting and filtering; carrying out rotary evaporation on filtrate to remove n-hexane to obtain red shrimp sauce; adding acetone into the red shrimp sauce for dissolving, and filtering to obtain acetone extract; carrying out rotary evaporation for removing acetone to obtain a red shrimp sauce sample containing the astaxanthin diester; dissolving the red shrimp sauce sample by petroleum ether for standby; taking a chromatographic column, packing, loading the red shrimp sauce sample and eluting to obtain a yellow strip; and carrying out rotary evaporation for removing solvent to obtain the astaxanthin diester. The method for extracting the astaxanthin diester from the antarctic krill adopts n-hexane and acetone for extracting and the column chromatography for preparing the astaxanthin diester, and is less in dosage of the solvent as well as simple and easy in technical process, thus providing technical reference for extracting the astaxanthin diester from the antarctic krill.
Description
Technical field
The present invention relates to a kind of method of extracting astaxanthin diester from krill.
Background technology
Krill (Euphausia superba), has another name called large krill or South Pole large krill, is the krill of one way of life in the waters, the Antarctica of Antarctic Ocean.Live in the mode of trooping, density reaches 10,000-30,000/m sometimes
3.They may be the most successful animal species on the earth, hundred million tons of nearly 5-50 (because of the methods of evaluation larger difference that differed).Annual approximately 100,000 tons of krill fishery.Mainly fishery country is Japan and Polish.Product is mainly used as cooking in Japan, and world's elsewhere is as animal-feed and bait.
In krill, contain the free astaxanthin of minute quantity, this be because astaxanthin mostly and the lipid acid formation astaxanthin ester that combines, astaxanthin ester is the main existence form of krill astaxanthin.When extract grease from krill time, astaxanthin ester is just extracted together with oil.
Astaxanthin ester is divided into again astaxanthin diester and astaxanthin one ester.Astaxanthin and the diester being formed by it and an ester are at the different times in life of krill, and difference is survived season, different living environments, and it is different between the content in its shrimp body and three, having ratio.Therefore one of the marker that can study three as krill growth metabolism, can be used as the marker that ripe shoal of shrimps catching season timing is fished for, environmental positioning is studied.Due to krill after fishing for through freezing or be dried to dry shrimp or dry powder astaxanthin ester in storage usually decomposes, so astaxanthin, the content of astaxanthin diester and an ester can be used as one of index of raw material freshness.
Astaxanthin ester is more stable than astaxanthin, and astaxanthin ester and astaxanthin have very strong antioxygenation.Therefore astaxanthin diester, quality and the shelf-life of astaxanthin one the ester ratio in shrimp sauce and shrimp sauce with astaxanthin are relevant.
But up to the present, people only separate and obtain astaxanthin ester and astaxanthin.Especially astaxanthin is produced.But not yet astaxanthin diester is separated from astaxanthin ester.The lipid acid of its astaxanthin diester institute combination of different types of shrimp is also different.Exclusive oneself the astaxanthin diester molecule colony of each seed shrimp.Parameter and method that the astaxanthin diester of different shrimps separates preparation also have larger difference.
Summary of the invention
For above-mentioned prior art, the invention provides a kind of method of extracting astaxanthin diester from krill, its technological process is simple.
The present invention is achieved by the following technical solutions:
A method of extracting astaxanthin diester from krill, step is as follows:
(1) get 1 kilogram of dry krill, add 5～7 liters of normal hexanes, stir and extract 1～2h, extracting liquid filtering, extracts repeatedly, extracts altogether merging filtrate 4～5 times;
(2) filtrate obtained above is rotated evaporative removal normal hexane at 25～35 ℃, obtains red shrimp sauce;
(3) to the acetone solution that adds 10 times of volumes in red shrimp sauce obtained above, filter to obtain acetone extract, at 25～35 ℃, rotate evaporative removal acetone, obtain the red shrimp sauce sample of astaxanthin-containing diester;
(4) by the appropriate petroleum ether dissolution of the red shrimp sauce sample of astaxanthin obtained above, for subsequent use;
(5) take 200～300 object column chromatography silica gels of 40～60g, at 100～120 ℃, activate 12～18h;
(6) in sherwood oil: ratio preparation mixed solution 300mL ethyl acetate=10:1～1.2(V/V), as elutriant;
(7) get chromatography column, with the mixed solution in step (6) and the dress of the silica gel wet method in step (5) post;
(8) get the sample loading of 1～3mL step (4);
(9) elutriant in use step (6) is by the speed wash-out of 80～100/min, first the colour band of the next lurid 1～2cm of wash-out, then there is the yellow band (astaxanthin diester band) of a 8～12cm to be eluted, connect sample with the test tube of 15mm × 150mm, under 25～30 ° of C, rotate evaporative removal solvent, obtain astaxanthin diester.Approximately complete this band of wash-out needs 1.5h.
The specification of described chromatography column is 2～3cm × 30cm.
The detection of the astaxanthin diester that aforesaid method obtains:
The astaxanthin diester of preparation is got to 1g and be dissolved in methyl alcohol: in mixing solutions methylene dichloride=3:1(V/V).Get 2 μ L at silica gel G F with the micro-sampling device of 10 μ L
254point sample on efficient thin layer plate, using normal hexane: acetone=7:2(V/V) in expansion cylinder, launch 6cm as developping agent.Thin layer chromatography scanner light show sample only has a point, some R
fvalue is 0.89.
The contained fatyy acids of astaxanthin diester that aforesaid method obtains:
Get the astaxanthin diester 5-10mg of preparation, add 1mL10%(percent by volume) vitriol oil-methanol solution, in 60 ° of C water-bath esterification 15min, add the vibration of 1mL normal hexane after cooling, stratification.Getting supernatant liquor analyzes for GC/MS.
Analytical results: astaxanthin diester contains following lipid acid: tetradecanoic acid, 14-methyl pentadecylic acid, palmitinic acid, oleic acid, elaidic acid, 8, vaccenic acid, trans-11-Octadecenoic acid, 9,12,15-punicic acid (linolenic acid), cis-11-eicosenoic acid, 7,10,13-hiragonic acid, EPA, DHA.Wherein to be up to that 38.465%, DHA takes second place be 20.465% to EPA content, moreover for 14-methyl pentadecylic acid be 6.142%.
The method of extracting astaxanthin diester from krill of the present invention, adopt normal hexane and acetone extraction, column chromatography to prepare astaxanthin diester, solvent load is few, and technological process is simple, for the extraction preparation of astaxanthin diester in krill provides Technical Reference.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated.
Embodiment 1
Get dry krill 100g, add 600mL normal hexane, stir extraction 2h and obtain extracting solution, extracting liquid filtering, more repeatedly extract 3 times, merge the filtrate of 4 times; Filtrate is removed normal hexane at 30 ℃ of rotary evaporations, obtains the red shrimp sauce of 5g; In shrimp sauce, add 50mL acetone to mix dissolving, quiet heavy, filter to get filtrate; Filtrate is removed acetone at 30 ℃ of rotary evaporations, obtains red oil sample, for subsequent use with appropriate petroleum ether dissolution.
Take 300 order silica gel 60g, 120 ℃ of activation 12h.By sherwood oil: ethyl acetate=10:1 volume ratio preparation mixed solution 300mL adopts wet method that silica gel is filled to post to the chromatography column of 3cm × 30cm; Get 2mL sample loading for subsequent use.
With the elutriant wash-out of sherwood oil: ethyl acetate=10:1,90/min of elution speed, is washing after the pale yellow colour band of lower 1 wide about 1cm, the yellow astaxanthin diester band of a visible 10cm, and 30 ℃ of steamings desolventize to obtain astaxanthin diester.
The detection of the astaxanthin diester that aforesaid method obtains:
The astaxanthin diester of preparation is got to 1g and be dissolved in methyl alcohol: in mixing solutions methylene dichloride=3:1(V/V).Get 2 μ L at silica gel G F with the micro-sampling device of 10 μ L
254point sample on efficient thin layer plate, using normal hexane: acetone=7:2(V/V) in expansion cylinder, launch 6cm as developping agent.Thin layer chromatography scanner light show sample only has a point, some R
fvalue is 0.89.
The contained fatyy acids of astaxanthin diester that aforesaid method obtains:
Get the astaxanthin diester 8mg of preparation, add the 1mL10% vitriol oil-methanol solution, in 60 ° of C water-bath esterification 15min, add the vibration of 1mL normal hexane after cooling, stratification.Getting supernatant liquor analyzes for GC/MS.
Analytical results: astaxanthin diester contains following lipid acid: tetradecanoic acid, 14-methyl pentadecylic acid, palmitinic acid, oleic acid, elaidic acid, 8, vaccenic acid, trans-11-Octadecenoic acid, 9,12,15-punicic acid (linolenic acid), cis-11-eicosenoic acid, 7,10,13-hiragonic acid, EPA, DHA.Wherein to be up to that 38.465%, DHA takes second place be 20.465% to EPA content, moreover for 14-methyl pentadecylic acid be 6.142%.
Embodiment 2
Get dry krill 100g, add 500mL normal hexane, stir extraction 1h and obtain extracting solution, extracting liquid filtering, more repeatedly extract 4 times, merge the filtrate of 5 times; Filtrate is removed normal hexane at 25 ℃ of rotary evaporations, obtains the red shrimp sauce of 4g; In shrimp sauce, add 40mL acetone to mix dissolving, quiet heavy, filter to get filtrate; Filtrate is removed acetone at 35 ℃ of rotary evaporations, obtains red oil sample, for subsequent use with appropriate petroleum ether dissolution.
Take 200 order silica gel 40g, 100 ℃ of activation 18h; Chromatography column is 2cm × 30cm, preparation sherwood oil: the mixed solution wet method dress post of ethyl acetate=10:1.1 volume ratio, get 1mL sample loading for subsequent use.
With the elutriant wash-out of sherwood oil: ethyl acetate=10:1.1, elution speed is 80/min, and washing the pale yellow colour band of lower Article 1 is 1cm, and the yellow band of Article 2 8cm is astaxanthin diester band, and 25 ℃ of steamings desolventize to obtain astaxanthin diester.
Embodiment 3
Get dry krill 100g, add 700mL normal hexane, stir extraction 1h and obtain extracting solution, extracting liquid filtering, more repeatedly extract 3 times, merge the filtrate of 4 times; Filtrate is removed normal hexane at 35 ℃ of rotary evaporations, obtains the red shrimp sauce of 6g; In shrimp sauce, add 60mL acetone to mix dissolving, quiet heavy, filter to get filtrate; Filtrate is removed acetone at 25 ℃ of rotary evaporations, obtains red oil sample, for subsequent use with appropriate petroleum ether dissolution.
Take 300 order silica gel 50g, 110 ℃ of activation 15h; Chromatography column is 3cm × 30cm, preparation sherwood oil: the mixed solution wet method dress post of ethyl acetate=10:1.2 volume ratio, get 1mL sample loading for subsequent use.
With the elutriant wash-out of sherwood oil: ethyl acetate=10:1.2, elution speed is 100/min, and first next pale yellow colour band of wash-out is 2cm, then washes the wide yellow astaxanthin diester band of next 12cm, and 30 ℃ of steamings desolventize to obtain astaxanthin diester.
Claims (2)
1. a method of extracting astaxanthin diester from krill, is characterized in that: step is as follows:
(1) get 1 kilogram of dry krill, add 5～7 liters of normal hexanes, stir and extract 1～2h, extracting liquid filtering, extracts repeatedly, extracts altogether merging filtrate 4～5 times;
(2) filtrate obtained above is rotated evaporative removal normal hexane at 25～35 ℃, obtains red shrimp sauce;
(3) to the acetone solution that adds 10 times of volumes in red shrimp sauce obtained above, filter to obtain acetone extract, at 25～35 ℃, rotate evaporative removal acetone, obtain the red shrimp sauce sample of astaxanthin-containing diester;
(4) by the appropriate petroleum ether dissolution of the red shrimp sauce sample of astaxanthin obtained above, for subsequent use;
(5) take 200～300 object column chromatography silica gels of 40～60g, at 100～120 ℃, activate 12～18h;
(6) in sherwood oil: the ratio preparation mixed solution 300mL of ethyl acetate=10:1～1.2, as elutriant;
(7) get chromatography column, with the mixed solution in step (6) and the dress of the silica gel wet method in step (5) post;
(8) get the sample loading of 1～3mL step (4);
(9) with the elutriant in step (6) by the speed wash-out of 80～100/min, the first next lurid colour band of wash-out, then has a yellow band to be eluted, and rotates evaporative removal solvent at 25～30 ℃, obtains astaxanthin diester.
2. a kind of method of extracting astaxanthin diester from krill according to claim 1, is characterized in that: the specification of described chromatography column is 2～3cm × 30cm.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103864656B (en) * | 2014-03-03 | 2016-05-11 | 山东师范大学 | The method of astaxanthin one ester of astaxanthin odd-numbered fatty acid one ester is rich in preparation |
| CN103820223B (en) * | 2014-03-03 | 2015-12-09 | 山东师范大学 | Be rich in saturated and be rich in the separation method of krill astaxanthin diester of unsaturated fatty acids |
| CN105085258B (en) * | 2015-07-23 | 2017-03-15 | 山东师范大学 | A kind of method for extracting dioctyl adipate from Antarctic krill |
| CN105067735B (en) * | 2015-07-23 | 2017-03-22 | 山东师范大学 | Method for extracting and detecting dioctyl adipate from Euphausia superba |
| CN109251156A (en) * | 2018-10-11 | 2019-01-22 | 中国水产科学研究院黄海水产研究所 | The preparation method of astaxanthin diester reference material in a kind of krill |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1089620A (en) * | 1993-12-15 | 1994-07-20 | 屈步华 | Preparation method and product with natural structure chitin |
| JP4063551B2 (en) * | 2002-02-18 | 2008-03-19 | 富士通株式会社 | Character string prediction apparatus and method, and computer-executable program for implementing the method |
| CN102041166A (en) * | 2011-01-20 | 2011-05-04 | 山东师范大学 | Method for extracting krill oil with high phosphatide content from Antarctic krills |
| CN102334686A (en) * | 2011-07-19 | 2012-02-01 | 中国水产科学研究院东海水产研究所 | Rapid low temperature extraction method for total astaxanthin from frozen Euphausia superba Dana |
Family Cites Families (1)
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| JPH0463551A (en) * | 1990-06-29 | 1992-02-28 | Daicel Chem Ind Ltd | Composition for improving body color or meat color of cultured animal and method for culturing animal |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1089620A (en) * | 1993-12-15 | 1994-07-20 | 屈步华 | Preparation method and product with natural structure chitin |
| JP4063551B2 (en) * | 2002-02-18 | 2008-03-19 | 富士通株式会社 | Character string prediction apparatus and method, and computer-executable program for implementing the method |
| CN102041166A (en) * | 2011-01-20 | 2011-05-04 | 山东师范大学 | Method for extracting krill oil with high phosphatide content from Antarctic krills |
| CN102334686A (en) * | 2011-07-19 | 2012-02-01 | 中国水产科学研究院东海水产研究所 | Rapid low temperature extraction method for total astaxanthin from frozen Euphausia superba Dana |
Non-Patent Citations (1)
| Title |
|---|
| 吴彩娟.天然虾青素提取和提纯工艺研究.《中国优秀博硕士学位论文全文数据库 (硕士)，工程科Ⅰ辑》.2003,第14页虾青素提取，第22页第（4）节柱层析和结晶. * |
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