70 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的和 DHP-EE % (RSD 1.05 %) respectively. The RSD of repetition was 1.54 %(n=6) for and 1.25 %(n=6) for. This method was

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1 DOI /j.issn Food Research And Development * : 采用超高压液相方法检测鱼油中的二十碳五烯酸乙酯 () 和二十二碳六烯酸乙酯 (), 采用 UPLC BEH C18(2.1 mm 50 mm,1.7 μm) 色谱柱, 考察了流动相有机相比例流速及柱温对鱼油中和 DHA- EE 分离的影响, 确定了最佳色谱条件 : 等度洗脱, 流动相为乙腈 - 水 (80 20, 体积比 ), 流速 0.5 ml/min, 柱温 30, 检测波长 220 nm 该方法下, 和分别在 10.4 μg/ml~209 μg/ml 10.3 μg/ml~207 μg/ml 范围内浓度和面积呈现良好的线性关系, 相关系数 r 分别为和 ; 和的方法的检测限为 4 ng 和 6 ng, 定量限分别为 10 ng 和 15 ng, 精密度试验 RSD(n=6) 分别为 1.09 % 和 0.34 %, 重复性试验 RSD(n=6) 分别为 1.54 % 和 1.25 %, 平均回收率分别为 %(RSD=1.33 %) 和 %(RSD=1.05 %) 本方法快速简单可靠灵敏重复性好, 可用于鱼油中和的快速检测 : 超高压高效液相色谱 ; 二十碳五烯酸乙酯 ; 二十二碳六烯酸乙酯 ; 鱼油 Determination of Eicosapentaenoic Acid Ethyl and Docosahexaenoic Acid Ethyl Ester in Fish Oil by Ultra Performance Liquid Chromatography CHEN Wei-zhu 1,2, JIN Wen-hui 1, HONG Zhuan 1,*, ZHANG Yi-ping 1, FANG Hua 1, YI Rui-zao 1 (1.The Third Institute of Oceanography State Oceanic Administration, Xiamen , Fujian, China; 2. Department of Chemistry, College of Chemistry and Chemical Engineering, and The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen , Fujian, China) Abstract The article is researching determination of eicosapentaenoic acid ethyl ester (EPA -EE) and docosahexaenoic acid ethyl ester (DHA- EE) in Fish Oil by Ultra performance liquid chromatography(uplc). The analysis was performed on an Acquity UPLC BEH C18 column (2.1 mm 50 mm, 1.7 μm). The influence of flow rate, the proportion of organic phase in mobile phase, and column temperature on the separation of and in fish oil was comprehensively studied. The optimal separation condition was as follows: a mobile phase consisting of acetonitrile and water (80 20,V/V) with a isocratic elution profile, UV detection wavelength at 220 nm, 0.5 ml/min of flow rate for mobile phase, 30 of column temperature. On the above condition, good linearity between response (peak area) and concentration was found over a concentration range of 10.4 μg/ml~209 μg/ml for, and 10.3 μg/ml~207 μg/ml for respectively. And the correlation coefficient of the calibration curve was for, and for respectively. The limits of detection (S/N>3) were 4ng for and 6ng for respectively. The limits of quantification (S/N>10) were 10ng for and 15ng for respectively. The average recovery rates of and were % (RSD 1.33 %) and 2009Y * 1970

2 70 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的和 DHP-EE % (RSD 1.05 %) respectively. The RSD of repetition was 1.54 %(n=6) for and 1.25 %(n=6) for. This method was simple, accurate and sensitive with a good reproducibility. It is suitable for fast detection of and in fish oil. Key words:ultra performance liquid chromatography ( UPLC ) ; eicosapentaenoic acid methyl ester ; docosahexaenoic acid ethyl ester; fish oil cis eicosapentaenoic acid EPA cis docosahexaenoic acid DHA ω-3 EPA DHA EPA DHA Milli-Q 1.3 [1-2] EPA DHA EPA -EE mg EPA DHA mg 25 ml DHA- EE EPA DHA EPA 1.2 DHA EPA DHA [3] 1.7 μm ml/min 220 nm 3 μl 30 [4-5] [6] 2 [7] 10 min μm UPLC - - UPLC - <2 μm >10 5 kpa 75 % 80 % 85 % 1 [8-13] ml/min TUV ACQUITY Waters ACQUITYUPLC TM BEHC mm 50 mm Sigma-Aldrich ACQUITY UPLC TM BEHC18 2.1mm 50 mm ml/min 1.7 μm Waters Milii-Q Millipore

3 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的和 DHP-EE a a 0.3 ml/min b c Fig.1 Chromatograms of and in different mobile phases with the same flow rate and the same column temperature ml/min 2.3 [4] 220 nm y= x y x 30 r = y= x y x 0.5 ml/min nm 2.4 EPA -EE r = S/ N μl 7 5 ml 3 4 ng 6 ng 7 S/ N ng 15 ng μl b 0.4 ml/min c 0.5 ml/min d 0.6 ml/min Fig.2 Chromatograms of and in the same mobile phases and the same column temperature with different flow rate

4 a b c 40 Fig.3 Chromatograms of and in the same mobile phases and the same flow rate with different column temperature Table 1 1 Calibration curve for and / μg/ml 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的和 DHP-EE 104 μg/ml 103 μg/ml 3 μl 6 RSD 1.09 % 0.34 % μg/ml 103 μg/ml 6 3 μl RSD 1.54 % 1.29 % EPA- EE μg % RSD 1.33 % % RSD 1.05 % 2 2 Table 2 The result of recovery for / / μg/ml μg/ml % /% /% EPA- EE DHA -EE GB/T [20] 3 Table 3 3 The content of and in Sample by two methods / mg/ml / mg/ml

5 [7],, EPA DHA 6 [8],, [J].,2010,29(9):61-64 UPLC [9] HPLC HPLC, 2011, 39(7): 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的和 DHP-EE [1],. [12],, [J]., 1998, 19(3):29-36 [2],,. n-3 27(12): [J]., 1997, 2(1):65-70 [3],,,. HPLC [J].,2010,21(13): [4] ALKIO M, JANTTI M, AALTONEN O. Purification of polyunsaturated fatty acid esters from tuna oil with supercritical fluid chromatography[j]. Journal of the american oil chemists society,2000,77(3): [5],,,. HPLC DHA [J]., 2011,36(2): [6],,,. / EPA /DHA [J].,2008,24(2): ,. [J]., 2010(4): [J]. [10],,,. [J].,2008,26(6): [11],,,. [J]. :,2011,47(3): , [J].,2008, [13] YANG F Q, GUAN J, LI S P. Fast simultaneous determination of 14 nucleosides and nucleobases in cultured Cordyceps using ultra - performance liquid chromatography[j].talanta, 2007(73): !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! CN /TS ISSN / tjfood@vip.163.com

DOI /j.issn Food Research And Development UPLC-MS/MS 5 0.

DOI /j.issn Food Research And Development UPLC-MS/MS 5 0. 118 217 5 38 1 DOI 1.3969/j.issn.5-6521.217.1.28 Food Research And Development 1 2 1 2 1 2 1 2 1 2 1 2 1. 731 2. 731 - UPLC-MS/MS 5. 1 mol/l ProElut PXC 5 % - Thermo Hypersil Gold mm 2.1 mm 1.9 μm ESI

70 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的 和 DHP-EE % (RSD 1.05 %) respectively. The RSD of repetition was 1.54 %(n=6) for and 1.25 %(n=6) for. This method was

70 陈伟珠, 等 : 超高压液相色谱法测定鱼油中的和 DHP-EE % (RSD 1.05 %) respectively. The RSD of repetition was 1.54 %(n=6) for and 1.25 %(n=6) for. This method was