固相萃取-反相高效液相色谱法测定蛋黄中的斑蝥黄  

Determination of Canthaxanthin in Egg Yolk Based on Solid-PhaseExtraction and Reversed-Phase High-PerformanceLiquid Chromatography Method

作　　者：王凤芹[1] 刘鑫[1] 程远之 郭冠群 路则庆[1] 汪以真[1] WANG Fengqin;LIU Xin;CHENG Yuanzhi;GUO Guanqun;LU Zeqing;WANG Yizhen(Key Laboratory of Animal Feed and Nutrition of Zhejiang Province,Key Laboratory of Animal Nutrition and Feed Science(Eastern of China),Ministry of Agriculture and Rural Affairs,Feed Science Institute of Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学饲料科学研究所,农业农村部(华东)动物营养与饲料重点实验室,浙江省饲料与动物营养重点实验室,杭州310058

出　　处：《动物营养学报》2021年第8期4675-4683,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：现代农业生猪产业体系(CARS-36);浙江省高校实验室工作研究项目(ZD202002)。

摘　　要：本试验旨在建立一种定量测定蛋黄中斑蝥黄的固相萃取-反相高效液相色谱法。蛋黄用乙腈在超声波辅助下提取,提取液经过Oasis PRiME HLB固相萃取柱净化,再由C 18色谱柱分离,流动相为乙腈-水(95∶5,体积比),流速为1.2 mL/min,等度洗脱,在紫外-可见吸收波长474 nm处检测,外标法定量。结果显示:蛋黄中斑蝥黄的检出限为0.05μg/g,定量限为0.1μg/g。斑蝥黄在0.1~10.0μg/mL的浓度范围内线性关系良好,相关系数(R^(2))为0.9999。在3个加标浓度(0.1、1.0和4.0μg/g)下,斑蝥黄的回收率为85.9%~98.6%,相对标准偏差(RSD)为1.7%~8.0%。应用上述方法测得市售新鲜鸡蛋蛋黄中斑蝥黄含量高达8.3μg/g。本试验建立的固相萃取-反相高效液相色谱法能够满足蛋黄中斑蝥黄及其异构体快速定量的需要。A simplified solid-phase extraction(SPE)and reversed-phase high-performance liquid chromatography(RP-HPLC)method for the quantitative determination of canthaxanthin in egg yolk was established in this experiment.The egg yolk sample was extracted using acetonitrile and cleaned up on Oasis PRiME HLB SPE cartridges,the separation was achieved on a C 18 column using acetonitrile and water(95∶5,v/v)at a flow rate of 1.2 mL/min of isocratic elution.Canthaxanthin was quantified by an external standard method coupled to a diode array detector at 474 nm.The results showed that the limit of detection was 0.05μg/g and the limit of quantification was 0.1μg/g.Within the range of canthaxanthin concentration at 0.1 to 10.0μg/mL,the linear relationship was good,and the correlation coefficient(R^(2))was 0.9999.Under the three spiked concentrations(0.1,1.0 and 4.0μg/g),the recovery of canthaxanthin was 85.9%to 98.6%,and the relative standard deviation(RSD)was 1.7%to 8.0%.Measured by the above method,the canthaxanthin content in egg yolk of commercial fresh eggs up to 8.3μg/g.The SPE and RP-HPLC method developed in this experiment can be used for the fast analysis of canthaxanthin isomers in egg yolk.

关 键 词：固相萃取 高效液相色谱 斑蝥黄 

分 类 号：O657.72[理学—分析化学]