He Limin, Huang Xianhui, Fang Binghu, Huang Shixin, Cao Ying, Chen Jianxin, Zeng Zhenling, Chen Zhangliu
Department of Pharmacology and Toxicology, College of Veterinary Medicine, South China Agriculture University, Guangzhou 510642, China.
Se Pu. 2008 Nov;26(6):714-9.
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 microg/kg for testosterone, methyltestosterone, boldenone, methandienone and stanozolol, and 0.4 microg/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 microg/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 microg/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.
建立了一种基于超高效液相色谱-串联质谱法（UPLC-MS/MS）同时测定动物肌肉组织和蛋类中11种甾体激素残留量的可靠方法。这11种甾体激素分别为睾酮、甲基睾酮、群勃龙、勃地酮、诺龙、美雄酮、司坦唑醇、孕酮、丙酸诺龙、丙酸睾酮和苯丙酸诺龙。样品在碱性pH条件下用叔丁基甲醚萃取，然后通过冷冻脂质过滤进行净化。所有这些药物都可以通过UPLC-MS/MS在10分钟内进行测定，采用电喷雾电离正离子模式和多反应监测模式。睾酮、甲基睾酮、勃地酮、美雄酮和司坦唑醇的检测限为0.3微克/千克，群勃龙、诺龙、孕酮、丙酸睾酮和苯丙酸诺龙的检测限为0.4微克/千克。在猪肉、牛肉、羊肉、鸡肉肌肉组织和蛋类中添加1、2和10微克/千克水平的睾酮、甲基睾酮、勃地酮、美雄酮和司坦唑醇，总体回收率为62.3% - 105%，相对标准偏差（RSD）为0.5% - 15%。群勃龙、诺龙、孕酮、丙酸睾酮和苯丙酸诺龙的回收率高于50.0%，RSD低于16%。每种药物的基质校准曲线在1至100微克/升范围内呈线性（r > 0.99）。所建立的方法简便、快速、灵敏且特异，适用于动物肌肉组织和蛋类中合成代谢雄激素类固醇的鉴定和定量。
