HPLC法测定禽蛋蛋黄中玉米黄质和叶黄素
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摘要
建立高效液相色谱(HPLC)快速准确检测禽蛋蛋黄中玉米黄质和叶黄素的方法。色谱柱SHIMADZU~?C_(18)柱(4.6 mm×250 mm, 5μm),流动相为V_(乙腈P)︰V_(甲醇)︰V_(二氯甲烷)=85︰10︰5,流速1 mL/min,检测波长450 nm,温度为室温。确定以甲醇、乙酸乙酯和正己烷为混合提取溶剂,经水洗、盐洗、碱洗和水洗,制备得到待测液。结果表明,玉米黄质和叶黄素分别在10～160μg/m L和15～240μg/m L内呈良好线性关系,相关系数分别为0.997和0.999;玉米黄质的平均加标回收率为97.8%, RSD为1.19%(N=6);叶黄素的平均加标回收率为98.2%, RSD为1.90%(N=6);玉米黄质和叶黄素的精密度(RSD)分别为1.44%和1.77%,检出限分别为0.094μg/m L和0.048μg/m L。建立的HPLC方法能快速准确检测草鸡蛋、饲料鸡蛋、茶叶鸡蛋、鹌鹑蛋、鸭蛋和鹅蛋中蛋黄的玉米黄质和叶黄素含量,其中草鸡蛋蛋黄中叶黄素和玉米黄质含量最高,分别为57.50 mg/kg和56.87 mg/kg。此次试验为评价禽蛋的营养价值提供重要参考。
A rapid and accurate method for the determination of zeaxanthin and lutein in various egg yolks by high performance liquid chromatography(HPLC) was established. Chromatographic condition was SHIMADZU~? C_(18) column(4.6 mm×250 mm, 5 μm) with V_(acetonitrile)︰V_(methanol)︰V_(dichloromethane)=85︰10︰5 as mobile phase, flow rate of 1 mL/min, detection wavelength of 450 nm and the room temperature. The mixture of methanol, ethyl acetate and n-hexane was used as extracted solution, and the extracting solutions of egg yolks were washed by water, salt, alkali and water, then used as tested solution.The zeaxanthin and lutein showed good linearities in the range of 10-160 μg/mL and 15-240 μg/mL, respectively. The correlation coefficients were 0.997 and 0.999, respectively. The average spiked recovery rate of zeaxanthin was 97.8%; RSD was 1.19%(N=6); The average spiked recovery rate of lutein was 98.2%; RSD was 1.90%(N=6). The precisions(RSD) of zeaxanthin and lutein were 1.44% and 1.77%, respectively, and the detection limits were 0.094 μg/mL and 0.048 μg/mL,respectively. The HPLC method could quickly and accurately separate and detect the content of zeaxanthin and lutein in the egg yolks of cottage eggs, feed eggs, tea eggs, quail eggs, duck eggs and goose eggs. The content of lutein and zeaxanthin in egg yolks of cottage eggs was highest at 57.50 mg/kg and 56.87 mg/kg, respectively, which provided an important reference for evaluating the nutritional value of eggs.
A rapid and accurate method for the determination of zeaxanthin and lutein in various egg yolks by high performance liquid chromatography(HPLC) was established. Chromatographic condition was SHIMADZU~? C_(18) column(4.6 mm×250 mm, 5 μm) with V_(acetonitrile)︰V_(methanol)︰V_(dichloromethane)=85︰10︰5 as mobile phase, flow rate of 1 mL/min, detection wavelength of 450 nm and the room temperature. The mixture of methanol, ethyl acetate and n-hexane was used as extracted solution, and the extracting solutions of egg yolks were washed by water, salt, alkali and water, then used as tested solution.The zeaxanthin and lutein showed good linearities in the range of 10-160 μg/mL and 15-240 μg/mL, respectively. The correlation coefficients were 0.997 and 0.999, respectively. The average spiked recovery rate of zeaxanthin was 97.8%; RSD was 1.19%(N=6); The average spiked recovery rate of lutein was 98.2%; RSD was 1.90%(N=6). The precisions(RSD) of zeaxanthin and lutein were 1.44% and 1.77%, respectively, and the detection limits were 0.094 μg/mL and 0.048 μg/mL,respectively. The HPLC method could quickly and accurately separate and detect the content of zeaxanthin and lutein in the egg yolks of cottage eggs, feed eggs, tea eggs, quail eggs, duck eggs and goose eggs. The content of lutein and zeaxanthin in egg yolks of cottage eggs was highest at 57.50 mg/kg and 56.87 mg/kg, respectively, which provided an important reference for evaluating the nutritional value of eggs.
引文
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[10]刘春菊,肖亚冬,张钟元,等.玉米黄质及其异构体的测定方法研究[J].现代食品科技, 2015, 31(11):313-318.
[11]赵艳,杨发树,张凤枰,等.反相高效液相色谱法测定巴沙鱼中的黄体素和玉米黄质[J].食品科学, 2012, 33(16):225-228.
[12]黄小春,吴灵英,邱银生,等.用高效液相色谱法快速测定蛋黄中的叶黄素[J].饲料广角, 2007, 13:37-39.
[13]康翠欣,朱静璇,汪芳芳,等.高效液相色谱法快速测定婴幼儿乳粉中叶黄素的含量[J].食品科技, 2017, 42(11):312-315.
[14]曾东慧.柑橘果皮类胡萝卜素提取物对鸡蛋蛋黄中类胡萝卜素成分及其色泽影响的研究[D].杭州:浙江师范大学, 2013.
[2] CHEW E Y, CLEMONS T E, AGRON E, et al. Effect of Omega-3 fatty acids, lutein/zeaxanthin, or other nutrient supplementationoncognitivefunctiontheAREDS2randomized clinical trial[J]. JAMA, 2015, 314(8):791-801.
[3] JOHNSON E J. Role of lutein and zeaxanthin in visual and cognitive function throughout the lifespan[J]. Nutrition Reviews, 2015, 72(9):605-612.
[4] CHEW E Y, CLEMONS T E, SANGIOVANNI J P, et al.Lutein+zeaxanthin and omega-3 fatty acids for age-related macular degeneration:the Age-Related Eye Disease Study 2(AREDS2)randomized clinical trial[J]. Journal of the American Medical Association, 2013, 309(19):2005-2015.
[5]米佳,闫亚美,秦垦,等.皂化对枸杞类胡萝卜素组成的影响[J].食品工业, 2017, 38(6):152-154.
[6] LE M, RONG L, HUI D J, et al. Lutein, zeaxanthin and meso-zeaxanthin supplementation associated with macular pigment optical density[J]. Nutrients, 2016, 8(7):426.
[7]刘美宏,刘回民,郑明珠,等.玉米黄素生物活性及其在食品中应用研究进展[J].食品工业, 2016, 37(12):242-248.
[8] GIORDANO E, QUADRO L. Lutein, zeaxanthin and mammaliandevelopment:Metabolism,functionsand implications for health[J]. Archives of Biochemistry and Biophysics, 2018:33-40.
[9]赵学超.蛋黄中叶黄素分离纯化及功能性质研究[D].无锡:江南大学, 2009.
[10]刘春菊,肖亚冬,张钟元,等.玉米黄质及其异构体的测定方法研究[J].现代食品科技, 2015, 31(11):313-318.
[11]赵艳,杨发树,张凤枰,等.反相高效液相色谱法测定巴沙鱼中的黄体素和玉米黄质[J].食品科学, 2012, 33(16):225-228.
[12]黄小春,吴灵英,邱银生,等.用高效液相色谱法快速测定蛋黄中的叶黄素[J].饲料广角, 2007, 13:37-39.
[13]康翠欣,朱静璇,汪芳芳,等.高效液相色谱法快速测定婴幼儿乳粉中叶黄素的含量[J].食品科技, 2017, 42(11):312-315.
[14]曾东慧.柑橘果皮类胡萝卜素提取物对鸡蛋蛋黄中类胡萝卜素成分及其色泽影响的研究[D].杭州:浙江师范大学, 2013.