高效液相色谱-串联质谱法检测配合饲料中11种蛋白同化激素含量  

Determination of 11 Anabolic Androgenic Steroids in Compound Feeds by High Performance Liquid Chromatography-Tandem Mass Spectrometry

作　　者：王博 张婧[1] 贡松松 张浩然 徐汀 陈思思[1] 吴剑平[1] 严凤[1] WANG Bo;ZHANG Jing;GONG Song-song;ZHANG Hao-ran;XU Ting;CHEN Si-si;WU Jian-ping;YAN Feng(Shanghai Animal Disease Prevention and Control Center,Shanghai 201103,China)

机构地区：[1]上海市动物疫病预防控制中心,上海201103

出　　处：《分析测试学报》2022年第12期1779-1784,共6页Journal of Instrumental Analysis

摘　　要：建立了快速分析配合饲料中睾酮、甲基睾酮、勃地龙、美雄酮、雄烯二酮、脱氢异雄酮、诺龙、丙酸诺龙、司坦唑醇、美伦孕酮、黄体酮11种蛋白同化激素的高效液相色谱-串联质谱(HPLC-MS/MS)法。样品采用乙腈提取,经PSA粉净化后上机测定。采用Phenomenex C_(18)(100 mm×2.1 mm,2.6μm)色谱柱,以0.01%甲酸溶液-乙腈作为流动相进行梯度洗脱,电喷雾离子源正离子模式检测,同位素内标法定量。结果表明,11种蛋白同化激素的线性范围为1~100 ng/mL,相关系数均为0.999,检出限为20μg/kg,定量下限为50μg/kg。50、250、500μg/kg加标水平下,鸡配合饲料中各蛋白同化激素的回收率为94.5%~111%,日内相对标准偏差(RSD)和日间RSD均不大于13%;猪配合饲料中各蛋白同化激素的回收率为90.1%~109%,日内RSD不大于9.0%,日间RSD不大于8.8%。实际样品中检出睾酮和勃地龙,含量分别为9.09~14.68 mg/kg和1.22~1.84 mg/kg。该方法可为饲料中蛋白同化激素的滥用监管提供技术支撑。A high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was established for the rapid analysis of 11 anabolic androgenic steroids,i.e.testosterone,17-me⁃thyltestosterone,boldenone,metandienone,androstenedione,dehydroepiandrosterone,nandro⁃lone,nandrolone 17-propionate,stanozolol,melengestrol,progesterone in compound feeds.The samples were extracted with acetonitrile,then purified with PSA,and finally determined on the ma⁃chine.The separation of target compounds was performed on an Phenomenex C_(18)(100 mm×2.1 mm,2.6μm)by gradient elution,using 0.01%formic acid-acetonitrile system as mobile phase.The 11 anabolic androgenic steroids were detected in positive ESI mode under multiple reaction monitoring(MRM)mode,and quantified by the isotope internal standard method.The results showed that the linear ranges for 11 anabolic androgenic steroids were 1-100 ng/mL,and the correlation coefficients were all 0.999.The limits of detection and the limits of quantitation were 20μg/kg and 50μg/kg,respectively.The recoveries for 11 anabolic androgenic steroids in chicken compound feed ranged from 94.5%to 111%,with intra-day relative standard deviations(intra-RSDs)and inter-day RSDs(inter-RSDs)both not more than 13%,while the recoveries for the analytes in pig compound feed ranged from 90.1%to 109%,with intra-RSDs and inter-day RSDs not more than 9.0%and 8.8%,re⁃spectively.Testosterone and boldenone were detected in actual samples with the contents of 9.09-14.68 mg/kg and 1.22-1.84 mg/kg,respectively.This method could realize the comprehensive,simple and accurate detection of anabolic androgenic steroids in compound feeds,and provide techni⁃cal support for the regulation of anabolic androgenic steroids abuse in compound feeds in the future.

关 键 词：蛋白同化激素 高效液相色谱-串联质谱法 配合饲料 内标法 

分 类 号：O657.7[理学—分析化学] S816.17[理学—化学]