CN102731358A - Method for extracting astaxanthin diester from antarctic krill - Google Patents
Method for extracting astaxanthin diester from antarctic krill Download PDFInfo
- Publication number
- CN102731358A CN102731358A CN2012102486782A CN201210248678A CN102731358A CN 102731358 A CN102731358 A CN 102731358A CN 2012102486782 A CN2012102486782 A CN 2012102486782A CN 201210248678 A CN201210248678 A CN 201210248678A CN 102731358 A CN102731358 A CN 102731358A
- Authority
- CN
- China
- Prior art keywords
- astaxanthin
- extracting
- diester
- astaxanthin diester
- acetone
- 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 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 from krill, extracting astaxanthin diester.
Background technology
Krill (Euphausia superba) has another name called large krill or South Pole large krill, is a kind of krill that lives in the waters, the Antarctica of Antarctic Ocean.Live with the mode of trooping, density reaches 10,000-30,000/m sometimes
3They possibly be animal species the most successful on the earth, hundred million tons of nearly 5-50 (because of methods of evaluation has differed than big-difference).Annual about 100,000 tons of krill fishery.Main fishery country is a Japan and Polish.Product mainly is used as cooking in Japan, and world's elswhere is then as animal-feed and bait.
The free astaxanthin that contains minute quantity in the krill, 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 from krill, extracting grease, astaxanthin ester just is extracted out with oil together.
Astaxanthin ester is divided into astaxanthin diester and astaxanthin one ester again.Astaxanthin and the diester that is formed by it and an ester be at the different times in life of krill, different existence season, and different living environments, it is different having ratio between intravital content of its shrimp and the three.Therefore can be with the three as one of marker of krill growth metabolism research, can be used as that ripe shoal of shrimps catching season is regularly fished for, the marker of environment Position Research.Because krill is being fished for after freezing or be dried to dried shrimp or dry powder astaxanthin ester in storage usually decomposes, so astaxanthin, the content of an 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 all have very strong antioxygenation.So astaxanthin diester, astaxanthin one ester is relevant with the quality and the shelf-life of the ratio of astaxanthin in shrimp sauce and shrimp sauce.
But up to the present, people only separate and obtain astaxanthin ester and astaxanthin.Especially astaxanthin is produced.But astaxanthin diester is not separated from astaxanthin ester as yet.Its astaxanthin diester institute bonded lipid acid 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 is separated preparation also have than big-difference.
Summary of the invention
To above-mentioned prior art, the invention provides a kind of method of from krill, extracting astaxanthin diester, its technological process is simple.
The present invention realizes through following technical scheme:
A kind of method of from krill, extracting astaxanthin diester, step is following:
(1) get 1 kilogram of dried krill, add 5~7 liters of normal hexanes, stir and extract 1~2h, extracting liquid filtering extracts repeatedly, extracts merging filtrate altogether 4~5 times;
(2) the above-mentioned filtrating that obtains is rotated the evaporative removal normal hexane down at 25~35 ℃, gets red shrimp sauce;
(3) in the above-mentioned red shrimp sauce that obtains, add the acetone solution of 10 times of volumes, cross and filter acetone extract, at 25~35 ℃ of rotation evaporative removal acetone down, the red shrimp sauce sample of astaxanthin-containing diester;
(4) the red shrimp sauce sample of the above-mentioned astaxanthin that obtains is used an amount of petroleum ether dissolution, subsequent use;
(5) take by weighing 200~300 purpose column chromatography silica gels of 40~60g, at 100~120 ℃ of following activation 12~18h;
(6) in sherwood oil: the ratio preparation mixed solution 300mL of ETHYLE ACETATE=10:1~1.2 (V/V), as elutriant;
(7) get chromatography column, with the dress of the silica gel wet method in mixed solution in the step (6) and the step (5) post;
(8) get appearance on the sample of 1~3mL step (4);
(9) with the speed wash-out of the elutriant in the step (6) by 80~100/min; The colour band of the next lurid 1~2cm of wash-out at first; There is the yellow band (astaxanthin diester band) of a 8~12cm to be eluted then; Test tube with 15mm * 150mm connects appearance, and rotation evaporative removal solvent promptly gets astaxanthin diester under 25~30 ° of C.Approximately wash-out is intact should need 1.5h by band.
The specification of said chromatography column is 2~3cm * 30cm.
The detection of the astaxanthin diester that aforesaid method obtains:
The astaxanthin diester of preparation is got 1g be dissolved in methyl alcohol: in the mixing solutions of 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 the efficient thin layer plate launches 6cm with normal hexane: acetone=7:2 (V/V) as developping agent in expansion cylinder.Thin layer chromatography scanner light show sample has only a point, some R
fValue is 0.89.
The astaxanthin diester institute fatty acids that aforesaid method obtains detects:
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, the cooling back adds the vibration of 1mL normal hexane, standing demix.Getting supernatant supplies GC/MS to analyze.
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), suitable-the 11-eicosenoic acid, 7; 10,13-hiragonic acid, EPA, DHA.Wherein EPA content is up to 38.465%, and it is 20.465% that DHA takes second place, moreover is that 14-methyl pentadecylic acid is 6.142%.
The method of from krill, extracting astaxanthin diester 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 of astaxanthin diester in krill preparation provides Technical Reference.
Embodiment
Below in conjunction with embodiment the present invention is further described.
Embodiment 1
Get dried krill 100g, add the 600mL normal hexane, stir extraction 2h and get extracting solution, extracting liquid filtering extracts 3 times more repeatedly, merges 4 times filtrating; Filtrating is removed normal hexane at 30 ℃ of rotary evaporations, gets the red shrimp sauce of 5g; In shrimp sauce, add the dissolving of 50mL acetone mixing, quiet heavy, cross and filter filtrating; Filtrating is removed acetone at 30 ℃ of rotary evaporations, gets the red oil sample, and is subsequent use with an amount of petroleum ether dissolution.
Take by weighing 300 order silica gel 60g, 120 ℃ of activation 12h.By sherwood oil: ETHYLE ACETATE=10:1 volume ratio preparation mixed solution 300mL adopts wet method that silica gel is adorned post to the chromatography column of 3cm * 30cm; Get appearance on the subsequent use sample of 2mL.
Use sherwood oil: the elutriant wash-out of ETHYLE ACETATE=10:1,90/min of elution speed, after washing the pale yellow colour band of 1 wide about 1cm, the xanchromatic astaxanthin diester band of a visible 10cm, 30 ℃ of steamings desolventize astaxanthin diester.
The detection of the astaxanthin diester that aforesaid method obtains:
The astaxanthin diester of preparation is got 1g be dissolved in methyl alcohol: in the mixing solutions of 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 the efficient thin layer plate launches 6cm with normal hexane: acetone=7:2 (V/V) as developping agent in expansion cylinder.Thin layer chromatography scanner light show sample has only a point, some R
fValue is 0.89.
The astaxanthin diester institute fatty acids that aforesaid method obtains detects:
Get the astaxanthin diester 8mg of preparation, add the 1mL10% vitriol oil-methanol solution, in 60 ° of C water-bath esterification 15min, the cooling back adds the vibration of 1mL normal hexane, standing demix.Getting supernatant supplies GC/MS to analyze.
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), suitable-the 11-eicosenoic acid, 7; 10,13-hiragonic acid, EPA, DHA.Wherein EPA content is up to 38.465%, and it is 20.465% that DHA takes second place, moreover is that 14-methyl pentadecylic acid is 6.142%.
Embodiment 2
Get dried krill 100g, add the 500mL normal hexane, stir extraction 1h and get extracting solution, extracting liquid filtering extracts 4 times more repeatedly, merges 5 times filtrating; Filtrating is removed normal hexane at 25 ℃ of rotary evaporations, gets the red shrimp sauce of 4g; In shrimp sauce, add the dissolving of 40mL acetone mixing, quiet heavy, cross and filter filtrating; Filtrating is removed acetone at 35 ℃ of rotary evaporations, gets the red oil sample, and is subsequent use with an amount of petroleum ether dissolution.
Take by weighing 200 order silica gel 40g, 100 ℃ of activation 18h; Chromatography column is 2cm * 30cm, the preparation sherwood oil: the mixed solution wet method dress post of ETHYLE ACETATE=10:1.1 volume ratio, get appearance on the subsequent use sample of 1mL.
Use sherwood oil: the elutriant wash-out of ETHYLE ACETATE=10:1.1, elution speed is 80/min, washing the pale yellow colour band of article one is 1cm, the yellow band of second 8cm is the astaxanthin diester band, 25 ℃ of steamings desolventize astaxanthin diester.
Embodiment 3
Get dried krill 100g, add the 700mL normal hexane, stir extraction 1h and get extracting solution, extracting liquid filtering extracts 3 times more repeatedly, merges 4 times filtrating; Filtrating is removed normal hexane at 35 ℃ of rotary evaporations, gets the red shrimp sauce of 6g; In shrimp sauce, add the dissolving of 60mL acetone mixing, quiet heavy, cross and filter filtrating; Filtrating is removed acetone at 25 ℃ of rotary evaporations, gets the red oil sample, and is subsequent use with an amount of petroleum ether dissolution.
Take by weighing 300 order silica gel 50g, 110 ℃ of activation 15h; Chromatography column is 3cm * 30cm, the preparation sherwood oil: the mixed solution wet method dress post of ETHYLE ACETATE=10:1.2 volume ratio, get appearance on the subsequent use sample of 1mL.
Use sherwood oil: the elutriant wash-out of ETHYLE ACETATE=10:1.2, elution speed are 100/min, and at first the pale yellow colour band of next bar of wash-out is 2cm, wash the wide xanchromatic astaxanthin diester band of next 12cm then, 30 ℃ of steamings desolventize astaxanthin diester.
Claims (2)
1. method of from krill, extracting astaxanthin diester, it is characterized in that: step is following:
(1) get 1 kilogram of dried krill, add 5~7 liters of normal hexanes, stir and extract 1~2h, extracting liquid filtering extracts repeatedly, extracts merging filtrate altogether 4~5 times;
(2) the above-mentioned filtrating that obtains is rotated the evaporative removal normal hexane down at 25~35 ℃, gets red shrimp sauce;
(3) in the above-mentioned redness that obtains, add the acetone solution of 10 times of volumes, cross and filter acetone extract,, must contain the red shrimp sauce sample of plain diester at 25~35 ℃ of rotation evaporative removal acetone down;
(4) the red shrimp sauce sample of the above-mentioned astaxanthin that obtains is used an amount of petroleum ether dissolution, subsequent use;
(5) take by weighing 200~300 purpose column chromatography silica gels of 40~60g, at 100~120 ℃ of following activation 12~18h;
(6) in sherwood oil: the ratio preparation mixed solution 300mL of ETHYLE ACETATE=10:1~1.2, as elutriant;
(7) get chromatography column, with the dress of the silica gel wet method in mixed solution in the step (6) and the step (5) post;
(8) get appearance on the sample of 1~3mL step (4);
(9) with the speed wash-out of the elutriant in the step (6) by 80~100/min, at first the next lurid colour band of wash-out has a yellow band to be eluted then, and rotation evaporative removal solvent promptly gets astaxanthin diester under 25~30oC.
2. a kind of method of from krill, extracting astaxanthin diester according to claim 1, it is characterized in that: the specification of said chromatography column is 2~3cm * 30cm.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210248678.2A CN102731358B (en) | 2012-07-18 | 2012-07-18 | Method for extracting astaxanthin diester from antarctic krill |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201210248678.2A CN102731358B (en) | 2012-07-18 | 2012-07-18 | Method for extracting astaxanthin diester from antarctic krill |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102731358A true CN102731358A (en) | 2012-10-17 |
CN102731358B CN102731358B (en) | 2014-05-14 |
Family
ID=46987748
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201210248678.2A Expired - Fee Related CN102731358B (en) | 2012-07-18 | 2012-07-18 | Method for extracting astaxanthin diester from antarctic krill |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102731358B (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103820223A (en) * | 2014-03-03 | 2014-05-28 | 山东师范大学 | Method for separating saturated fatty acid and unsaturated fatty acid from astaxanthin of euphausia superba |
CN103864656A (en) * | 2014-03-03 | 2014-06-18 | 山东师范大学 | Method for preparing astaxanthin monoester rich in astaxanthin odd-numbered fatty acid monoester |
CN105067735A (en) * | 2015-07-23 | 2015-11-18 | 山东师范大学 | Method for extracting and detecting dioctyl adipate from Euphausia superba |
CN105085258A (en) * | 2015-07-23 | 2015-11-25 | 山东师范大学 | Method for extracting dioctyl adipate from euphausia superba dana |
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 |
---|---|---|---|---|
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 |
CN1089620A (en) * | 1993-12-15 | 1994-07-20 | 屈步华 | Preparation method and product with natural structure chitin |
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)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
-
2012
- 2012-07-18 CN CN201210248678.2A patent/CN102731358B/en not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
CN1089620A (en) * | 1993-12-15 | 1994-07-20 | 屈步华 | Preparation method and product with natural structure chitin |
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》, 20 July 2003 (2003-07-20) * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103820223A (en) * | 2014-03-03 | 2014-05-28 | 山东师范大学 | Method for separating saturated fatty acid and unsaturated fatty acid from astaxanthin of euphausia superba |
CN103864656A (en) * | 2014-03-03 | 2014-06-18 | 山东师范大学 | Method for preparing astaxanthin monoester rich in astaxanthin odd-numbered fatty acid monoester |
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 |
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 |
CN105067735A (en) * | 2015-07-23 | 2015-11-18 | 山东师范大学 | Method for extracting and detecting dioctyl adipate from Euphausia superba |
CN105085258A (en) * | 2015-07-23 | 2015-11-25 | 山东师范大学 | Method for extracting dioctyl adipate from euphausia superba dana |
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 |
Also Published As
Publication number | Publication date |
---|---|
CN102731358B (en) | 2014-05-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102746205B (en) | Method for extracting astaxanthin monoester from antarctic krill | |
CN102731358B (en) | Method for extracting astaxanthin diester from antarctic krill | |
Gardner et al. | The component fatty acids of the lipids of some species of marine and freshwater molluscs | |
CN102041166B (en) | Method for extracting krill oil with high phosphatide content from Antarctic krills | |
Charrouf et al. | Ethnoeconomical, ethnomedical, and phytochemical study of Argania spinosa (L.) Skeels | |
Sakthivel et al. | Evaluation of physicochemical properties, proximate and nutritional composition of Gracilaria edulis collected from Palk Bay | |
US6358542B2 (en) | Antioxidant compositions extracted from olives and olive by-products | |
Djenontin et al. | Composition of Azadirachta indica and Carapa procera (Meliaceae) seed oils and cakes obtained after oil extraction | |
Culkin et al. | The fatty acids of some marine crustaceans | |
Ciriminna et al. | A circular economy approach to fish oil extraction | |
CN108179053A (en) | A kind of preparation method of high quality Antarctic krill oil | |
CN103320217A (en) | Method for extracting krill oil rich in phospholipid from euphausia superba | |
KR102222043B1 (en) | Method of krill oil preparation and composition of krill oil | |
CN103641717B (en) | A kind of method from extraction and isolation chlorogenic acid flowering period floral disc of sunflower into | |
CN106753775B (en) | Extraction method of antarctic krill oil with high astaxanthin content and high astaxanthin ester content | |
US9763897B2 (en) | Therapeutic astaxanthin and phospholipid composition and associated method | |
Lima Neto et al. | Volatile compounds and palynological analysis from pollen pots of stingless bees from the mid-north region of Brazil | |
CN102731361A (en) | Method for rapidly enriching astaxanthin and ester thereof from Euphausia superba | |
CN102731360B (en) | Method for extracting astaxanthin from antarctic krill | |
Marzorati et al. | Green extraction strategies for sea urchin waste valorization | |
RU2339387C1 (en) | Method for obtaining of biologically active additive from ascidium | |
Johnson et al. | Lipids from conidia of Erysiphe graminis tritici (powdery mildew) | |
Siahaan et al. | Antioxidant activity of two edible Korean seaweed oil obtained from SC-CO2 and solvent extraction | |
CN102746204B (en) | Method for using VE powder to quickly adsorb astaxanthin of antarctic krill and esters of astaxanthin and for VE separation | |
JP5613872B2 (en) | Method for producing lipids containing useful fatty acid residues such as docosapentaenoic acid and arachidonic acid |
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 | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140514 Termination date: 20150718 |
|
EXPY | Termination of patent right or utility model |