动物肌肉组织中甾类同化激素多组分残留的液相色谱-质谱检测方法  

Determination of Anabolic Hormones Multi-residues in Animal Muscle Tissues Using Liquid Chromatography-Tandem Mass Spectrometry

作　　者：秦燕[1] 陈捷[1] 张美金[1] 

机构地区：[1]广州出入境检验检疫局食品实验室,广州510623

出　　处：《分析化学》2006年第3期298-302,共5页Chinese Journal of Analytical Chemistry

基　　金：国家科技部"十五"科技攻关项目(No.2001BA804A11)基金资助

摘　　要：建立了不同动物肌肉中甾类同化激素（表睾酮、丙酸睾酮、19-去甲基睾酮、甲基睾酮、孕酮、甲羟孕酮、雌二醇、雌三醇、炔雌醇、雌酮）多残留量的LC/MS/MS确证方法.样品加醋酸缓冲溶液均质,加酶溶液酶解后,再加甲醇超声提取,用叔丁基甲醚液-液萃取至少2次,之后经反相固相萃取柱净化,以乙腈-水为流动相,经C18柱分离后进行LC/MS/MS多反应监测模式下的定性及定量分析.其中雄激素,孕激素采用正离子扫描,雌激素则采用负离子扫描.17beta-NT、MTS、ETS、MED、PG、PTS、EST、17beta-ES、EES和ESN的定量检出限为0.5～1.0 μg/kg.在1.0μg/kg的定量检出限添加水平,上述10种激素的平均回收率为55%～77%;相对标准偏差为7.1%～35%.可实现样本灵敏、准确的定性定量分析.A liquid chromatography-tandem mass spectrometric （LC/MS/MS） method was developed for the simultaneous identification and determination of muhi-residues amounts of steroid anabolic hormones epitestosterone （ETS）, testosterone 17-propionate （PTS）, androlone （ 17β-NT ）, 17α-methyltestosterone （MTS）, progesterone （PG）, medroxyprogesterone（MED）, 17β-estradiol （17β-ES）, 17α-ethinylestradiol （EES） and estrone （ESN） in the muscle tissues of various animal species. Homogenized tissue samples were enzymatically digested in acetate buffer （ pH 5.0）, then methanol was added and the mixtures were extracted under ultrasonication incubation. Clean-up was carded out with at least two times tertiary butyl methyl ether （MTBE） liquid-liquid partitioning followed by a further reverse phase solid phase extraction （SPE） cartridge purification. After C18 LC gradient elution separation using acetonitrile-water as mobile phase, the eluents were qualitatively and quantitatively determined under multi-reaction monitoring （MRM） scan type with tandem mass analyzer using positive polarity mode for the androgen and progestational hormones, negative polarity for the estrogen respectively. The limit of quantitation （LOQ） for the above 10 hormones were in the range of 0. 5- 1.0μg/kgo At the 1.0μ g/kg LOQ spiked level, the mean recoveries were in the range of 55% and 77%, and accordingly, the relative standard deviation range were in the range of 7.1% and 35%. The real sample test showed this method could be used for the sensitive and accurate determination of multi-steroid anabolic hormones residues in biological muscle samples.

关 键 词：甾类同化激素 多残留分析 反相固相萃取 液相色谱-质谱联用 

分 类 号：O657.63[理学—分析化学]