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1 7 Vol. 7 No. 06 Journal of Food Safety and Quality Feb., 06 EPA 翟 * 量, 沈立荣 (, 0058) 摘要 : 目的 EPA (Nannochloropsis sp.), EPA, 方法, 000 r/min 5 min, 05 结果 : 50, 0 min, 5:(mL/g),, 7.6%, EPA 9.0% 结论 EPA, EPA 关键词 : ; ; ; Optimization of the solvent extraction with ultrasonic assistance and fatty acid analysis of the enriched EPA ZHAI Liang, SHEN Li-Rong * (Department of Food Science and Nutrition, Zhejiang University, Hangzhou 0058, China) ABSTRACT: Objective To optimize the extraction method of eicosapentaenoic acid by solvent extraction with ultrasonic assistance, with Nannochloropsis sp. riched in EPA as raw materials, and analyze the composition of fatty acid of the extracted. Methods Samples were added at a certain ratio of liquid extraction solvent, and extracted for a certain time with ultrasonic assistance after thorough mixing. Then the mixed samples were centrifuged at speed of 000 r/min with 5 min. At last, the supernatant were dried via rotary evaporation method and then were dried at 05. The extracted microalgae oil s fatty acid compositions were analyzed by gas chromatography after methyl esterification. Results The optimal extraction conditions were as follows: temperature 50, extraction time 0 min, liquid-solid ratio 5:, times of extraction. The extraction rate of reached 7.6% under the optimal extraction conditions. Gas chromatography analysis showed that EPA, the main component of the was 9.0% in total fatty acids. Conclusion The proposed method is simple, and can get high extraction rate of and high content of EPA, which is suitable for extracting from Nannochloropsis sp. The technology will provide a scientific basis for industrialization of of EPA. KEY WORDS: eicosapentaenoic acid; Nannochloropsis sp; ; solvent extraction * 通讯作者 :,,, shenlirong@zju.edu.cn *Corresponding author: SHEN Li-Rong, Professor, Department of Food Science and Nutrition in School of Biosystems Engineering & Food Science of Zhejiang University, No.866, Yuhangtang Road, Xihu District, Hangzhou 0058, China. shenlirong@zju.edu.cn

2 80 7 引言 (eicosapentaenoic acid, EPA) n-,,,, [,], EPA n- (docosapentaenoic acid, DHA) ( ), EPA DHA, n- n- [-6], (Nannochloropsis oculata) EPA [7] (Schizochytrium sp.) (Ulkenia amoeboida) (Crypthecodinium cohnii) DHA 00, [8,9],,, [0,] EPA, [] [], CO [4-6] DHA EPA DHA n-,, EPA, EPA EPA,, EPA 材料与方法. 材料与仪器 ( ); - (Sigma ); (, ); (, ) BS-4S ( ( ) ); SK0G ( ); KA-000 ( ); RE-5AAB ( ); K0 ( ); DGX-905B- ( ); GC-4C ( ); ( ). 实验方法.. 提取工艺流程 0.5 g,,, 000 r/min 5 min,, 05, 000 r/min ( ).. 单因素试验 75% 90%, 9 : 60% 65% 70% 75% 80% 85% 85% 90% 95%; 5 : ; 8 : min ; 4 : 5: 0: 5: 0:; 4 : 4,.. 正交试验,, L 9 ( 4 ), [6]..4 微藻油得率测定 /%=( / ) 00%,..5 脂肪酸分析 [7], μl

3 , : EPA 8 : DB- (60 m 0.5 mm, 0.5 μm) 50 ml/min, 75 ml/min, 40, (FID) 70, 70 : 40 ( min), 6 /min, 70 ( min), 9 /min, 0 (8 min), /min, 0 ( min) N00..6 统计与分析, SAS 9. 结果与分析. 单因素实验.. 溶剂对微藻油得率的影响, 75% 90%, ( ), (P 0.0),,, [8], Fig. Effects of different concentrations of ethanol on the extraction rate of.. 浸提温度对微藻油得率的影响 ( ):, ; 50, ; 5~50, ; 55,, Fig. Effects of different solvents on extraction rate of.. 乙醇浓度对微藻油得率的影响,,, 60%~95% ( ), 60%~90%, ;,, 85% 90% 95% Fig. Effects of temperatures on the extraction rate of..4 浸提时间对微藻油得率的影响 ( 4), ~0 min, ; 0 min, %,,, 0 min

4 8 7 Fig. 4 4 Effects of ultrasonic time on the extraction rate of..6 提取次数对微藻油得率的影响,,, 90% 0: 50 0 min, ~4 ( 6),,,..5 液料比对微藻油得率的影响 ( 5),,,,,,, : 5: 0: 5: Fig. 6 6 Effects of extraction times on extraction rate of Fig. 5 5 Effects of the ratio of liquid to material on extraction rate of. 正交实验,, L 9 ( 4 ) ( ), : (D) (C) (B) (A), (P 0.000) A B C D, 90% 50 5:, 7.6% Table 表正交实验因素水平 Factors and levels of orthogonal test A (%) B ( ) C (ml/g) D

5 , : EPA 8 Table 表正交实验结果与分析 Analysis of orthogonal experimental results A B C D (%) 6.44± ± ± ± ± ± ± ± ±0.5 K K K k k k R Table 表正交实验结果方差分析 Variance analysis of orthogonal-test results F A B C D 脂肪酸分析 ( 7 4), EPA (AA) 7, EPA 9.0%, 4 结论, EPA : 90%, 5:, 50, 7.6%, EPA 9.0%, EPA

6 84 7 Fig.7 7 Gas chromatogram of 表 4 粗藻油脂肪酸组成 Table 4 The fatty acid compositions of (min) (%) AA EPA : AA: ; EPA: 参考文献 [] Swanson D, Block R, Mousa SA. Omega- fatty acids EPA and DHA: health benefits throughout life [J]. Adv Nutr, 0, (): 7. [] Connor WE. Importance of n- fatty acids in health and disease [J]. Am J Clin Nutr, 000, 7(S): 7S 75S. [] Gianpaolo A, Ugo N. Supercritical fluid extraction of bioactive lipids from the microalga Nannochloropsis sp. [J]. Eur J Lipid Sci Technol, 005, 07(6): [4],,. EPA DHA[J]., 00, 6(4): Hu AJ, Qiu TQ, Liang HH. Production of EPA and DHA by Marine Algae [J]. China Oils Fats, 00, 6(4): [5]. EPA DHA [J]., 000, 4(): Liang Y. Current status and prospect of studies on microalgal EPA and DHA [J]. J Fish China, 000, 4(): [6]. Nannochloropsis sp. [D]. :, 004. Wu Q. The primary research of microalgae Nannochloropsis sp. culture and extraction of its PUFAs [D]. Beijing: Beijing University of Chemical Technology, 004. [7],,,. N/P [J]., 000, 4(7): Wei D, Zhang XC, Sui ZH, et al. Nitrogen and N/P s effect on microspheres algae growth, total fat content and fatty acid composition in nannochloropsis oculata [J]. Marine Sci, 000, 4(7): [8],,,. DHA [J]., 0, 9(7): Huang WF, Zheng XH, Zeng YP, et al. Effect of DHA algal oil on the quality of yoghurt [J]. Mod Food Sci Technol, 0, 9(7): [9],,,. DHA [J]., 00, (): Wen XX, Li JP, Hou WW, et al. The nutrition and health

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Microalgae for oil: Strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor [J]. Biotechnol Bioeng, 009, 0(): 00. [],,. [J]., 005, 7(5): 56 6.

7 , : EPA 85 protection function and the application in food industry of the microalgal docosahexaenoic acid [J]. Food Sci, 00, (): [0] Moazami N, Ashori A, Ranjbar R, et al. Large-scale biodiesel production using microalgae biomass of Nannochloropsis [J]. Biores Technol, 0, 9(SI): [] Rodolfi L, Zittelli GC, Bassi N, et al. Microalgae for oil: Strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor [J]. Biotechnol Bioeng, 009, 0(): 00. [],,. [J]., 005, 7(5): Zhang Y, Li XM, Wang BG. Review and prospect of sea weed derived bioactive substances [J]. World Sci-Technol Res Dev, 005, 7(5): [],. [J]. Han J, Wei DX, Yun ZH. Lipid extraction by using low poison solvent [J]. Chin J Anal Chem, 00, 0(4): [6] Wang LJ, Weller CL. Recent advances in extraction of nutraceuticals from plants [J]. Trends Food Sci Technol, 006, 7(6): 00. [7] Shen LR, Lai CQ, Feng X, et al. Drosophila lacks C0 and C PUFAs [J]. J Lipid Res, 00, 5(0): [8],,. [J]., 0, 7(5): 0. Du XF, Zou N, Sun DH. Optimization on oil extraction from Nannochloropsis oculata droop [J]. China Oils Fats, 0, 7(5): 0. ( 责任编辑 : 杨翠娜 ) 作者简介, 00, (8): 5 6. Zhou QC, Zhao HC. Application of algae in giant freshwater shrimp feed [J]. Feed Res, 00, (8): 5 6. [4],. [J]. 翟量, 硕士研究生, 主要研究方向为食品安全与营养 ybzan0000@6.com, 006, 7(): 8 0. Tan CL, Yuan YJ. The application of aqueous enzymatic method in the extraction of plant oil [J]. Food Res Dev, 006, 7(): 8 0. [5],,. [J]., 沈立荣, 教授, 博士生导师, 主要研究方向为食品安全与营养 shenlirong@zju.edu.cn 00, 0(4):

Microsoft Word 聂雪梅.doc

Microsoft Word 聂雪梅.doc 4 4 Vol. 4 No. 4 2013 8 Journal of Food Safety and Quality Aug., 2013 聂雪梅 *, 李立, 孙利, 高飞 (, 100123) 摘要 : 2011 12,, 2007~2011,,,, 关键词 : ; ; ; ; Trade and notification situation on food import and export