色谱

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超高效液相色谱-串联质谱法测定动物肌肉组织和鸡蛋中残留的11种甾体激素类药物

贺利民1,黄显会1,方炳虎1,黄士新2,曹莹2,陈建新1,曾振灵1,陈杖榴1   

  1. 1.Department of Pharmacology and Toxicology, College of Veterinary Medicine, South China Agriculture University, Guangzhou 510642, China; 2.Shanghai Institute for Veterinary Drug and Feedstuffs Control, Shanghai 201103, China
  • 收稿日期:2008-05-17 修回日期:2008-08-17 出版日期:2008-11-30 发布日期:1983-06-25
  • 通讯作者: 曾振灵
  • 基金资助:
    农业部行业标准基金项目(070106-69)

Determination of eleven steroid hormones in animal muscle tissues and eggs using ultra-performance liquid chromatography-tandem mass spectrometry

HE Limin1, HUANG Xianhui1, FANG Binghu1, HUANG Shixin2, CAO Ying2, CHEN Jianxin1, ZENG Zhenling1, CHEN Zhangliu1   

  1. 1.Department of Pharmacology and Toxicology, College of Veterinary Medicine, South China Agriculture University, Guangzhou 510642, China; 2.Shanghai Institute for Veterinary Drug and Feedstuffs Control, Shanghai 201103, China
  • Received:2008-05-17 Revised:2008-08-17 Online:2008-11-30 Published:1983-06-25
  • Contact: ZENG Zhenling

摘要: 建立了采用超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定猪、牛、羊和鸡肌肉组织及鸡蛋中睾酮、甲基睾酮、黄体酮、群勃龙、勃地龙、诺龙、美雄酮、司坦唑醇、丙酸诺龙、丙酸睾酮及苯丙酸诺龙等11种甾体激素多残留的分析方法。试样在碱性条件下用叔丁基甲醚提取,冷冻离心脱脂净化,以乙腈和甲酸水溶液为流动相,梯度洗脱,反相液相色谱分离。采用电喷雾离子化、多反应监测方式(MRM),对11种甾体激素同时进行定性定量测定。动物肌肉和鲜蛋中睾酮、甲基睾酮、勃地龙、美雄酮及司坦唑醇的检出限为0.3 μg/kg,群勃龙、诺龙、黄体酮、丙酸诺龙、丙酸睾酮及苯丙酸诺龙的检出限为0.4 μg/kg。在动物组织及鸡蛋中添加1,2及10 μg/kg 水平的药物回收试验中,睾酮、甲基睾酮、勃地龙、美雄酮及司坦唑醇的回收率均在62.3%~105%之间,相对标准偏差为0.5%~15%;群勃龙、诺龙、黄体酮、丙酸诺龙、丙酸睾酮及苯丙酸诺龙的回收率大于50.0%,相对标准偏差小于16%。11种甾体激素在1~100 μg/L范围内,线性关系良好,相关系数都大于0.99。该方法的样品前处理简单、快速,测定灵敏、准确,选择性好,可满足动物源食品中甾体激素类药物多残留的同时测定。

关键词: 残留, 超高效液相色谱-串联质谱, 动物肌肉组织, 鸡蛋 , 甾体激素

Abstract: A credible method was developed for the simultaneous determination of eleven steroid hormone residues in animal muscle tissues and eggs based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The eleven steroid hormones were testosterone, methyltestosterone, trenbolone, boldenone, nandrolone, methandienone, stanozolol, progesterone, nadrolone propionate, testosterone propionate and nadrolone phenylpropionate. The samples were extracted with tert-butyl methyl ether at alkaline pH and then cleaned up by freezing-lipid filtration. All these drugs can be assayed in 10 min by UPLC-MS/MS using electrospray ionization in positive ion mode and multiple reaction monitoring mode. The limits of detection were 0.3 μg/kg for testosterone, methyltestosterone, boldenone, methandienone and stanozolol, and 0.4 μg/kg for trenbolone nandrolone, progesterone, testosterone propionate and nadrolone phenylpropionate. Overall recoveries of testosterone, methyltestosterone, boldenone, methandienone and stanozolol were 62.3%-105% from pork, beef, mutton and chicken muscle tissues, and eggs fortified at the 1, 2 and 10 μg/kg levels , and the relative standard deviations (RSDs) were 0.5%-15%. The recoveries of trenbolone nandrolone, progesterone, testosterone propionate and nadrolone phenylpropionate were higher than 50.0%, and the RSDs were lower than 16%. The matrix calibration curve for each drug was linear (r>0.99) from 1 to 100 μg/L. The established method is simple, rapid, sensitive and specific, and is appropriate for the identification and quantification of anabolic androgenic steroids in animal muscle tissues and eggs

Key words: animal muscles, eggs , residues, steroid hormones, ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)