作者：尤亮亮¹, 魏巍², 刘海燕¹, 王欣璐¹, 曾丽萍¹, 张玲琳¹, 聂远洋¹

作者单位：1. 新希望乳业控股有限公司技术中心, 四川 成都 610016;
2. 哈尔滨工业大学化工学院, 黑龙江 哈尔滨 150006

关键词：性激素;牛奶;固相萃取;高效液相色谱-质谱法

摘要：

采用固相萃取-高效液相色谱质谱联用法(SPE-HPLC/MS),建立牛奶中9种性激素(雌酮、雌三醇、17-雌二醇、诺龙、甲睾酮、丙酸睾酮、醋酸氯地孕酮、醋酸甲地孕酮、醋酸甲羟孕酮)残留的检测方法。样品经甲醇提取,过固相萃取柱净化,氮气吹干,残留物甲醇溶解后测定。其中雌激素(雌酮、雌三醇、17-雌二醇)采用负模式;其余性激素为正模式,进行多反应监测(MRM)模式定性定量分析。雌激素检出限(LODs)为1.1~1.2 g/L,定量限(LOQs)为3.63~3.96 g/L;其余性激素检出限(LODs)为0.1~0.5 g/L,定量限(LOQs)为0.33~1.65 g/L;分别在20.0~500.0 g/L及2.5~100.0 g/L范围内线性关系良好(r20.9944)。在50.0~100.0 g/L的添加水平上,9种性激素的平均回收率在82.7%~98.2%之间,变异系数(CV)为4.2%~11.2%。该法操作简单、灵敏度高,可用于牛奶中9种性激素的测定。

Solid-phase extraction followed by high-performance liquid chromatography tandem mass spectrometry for the determination of 9 hormones residues in milk

YOU Liangliang¹, WEI Wei², LIU Haiyan¹, WANG Xinlu¹, ZENG Liping¹, ZHANG Linglin¹, NIE Yuanyang¹

1. Technology Center, New Hope Dairy Holdings Co., Ltd., Chengdu 610016, China;
2. Institute of Chemical Industry, Harbin Institute of Technology, Harbin 150006, China

Abstract: A sensitive solid-phase extration/high-performance liquid chromatography-mass spectrometry method was established to determine 9 hormones residues(E1, E3, αE2, NT, MTS, PTS, CMA, MA, MPA) in milk. Milk samples were extracted first and then further purified on a solid-phase extraction column. After that, the purified solution was dried through nitrogen and the residues were dissolved in the mixture of methanol before LC-MS/MS analysis. Negative ion modes were adopted for 3 estrogens(E1, E3, αE2) while positive ion modes for all the other sex hormones. A multiple-reaction monitoring (MRM) mode was used for analysis. For the estrogens, the limits of detection (LODs) ranged from 1.1 μg/L to 1.2 μg/L whereas the limits of quantification (LOQs) were between 3.63 μg/L and 3.96 μg/L. For the other sex hormones, the LODs ranged from 0.1 μg/L to 0.5 μg/L whilst the LOQs were between 0.33 μg/L and 1.65 μg/L. Regression equations of these hormones had a good linear relationship(r² greater than 0.9944) within 20.0-500.0 μg/L and 2.5-100.0 μg/L. The average recoveries of 9 hormones were from 82.7% to 98.2% with the coefficients of variation(CV) between 4.2% and 11.2%(n=6) at the spiked levels of 50.0-100.0 μg/L. The method is simple, sensitive and therefore can be used to determine the residues of all the 9 hormones in cow milk.

Keywords: sex hormone;milk;solid-phase extraction(SPE);high-performance liquid chromatography-mass spectrometry(HPLC/MS)

2016, 42(5): 46-49,55  收稿日期: 2015-12-29;收到修改稿日期: 2016-02-02

基金项目: 国家科技支撑计划课题(2012BAD12B04)

作者简介: 尤亮亮(1988-),男,黑龙江绥化市人,硕士,主要从事乳品科学方面的研究。

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