张爱芝, 王全林＊, 沈 坚, 张书芬, 陈立仁

(1.宁波市产品质量监督检验研究院,浙江宁波315041;2.中国科学院兰州化学物理研究所,甘肃兰州730000)

超高效液相色谱-串联质谱法同时测定鱼制品中残留的7种性激素

张爱芝1, 王全林1＊, 沈 坚1, 张书芬1, 陈立仁2

(1.宁波市产品质量监督检验研究院,浙江宁波315041;2.中国科学院兰州化学物理研究所,甘肃兰州730000)

以电喷雾离子源(ESI)为电离源,在正离子采集模式下建立了鱼制品中7种性激素(甲基炔诺酮、甲基睾酮、丙酸睾酮、醋酸甲羟孕酮、醋酸甲地孕酮、醋酸氯地孕酮、诺龙)的超高效液相色谱-质谱/质谱(UPLC-MS/MS)检测方法。样品被酶解后用甲醇提取,提取液经氯化锌(ZnCl2)去脂、LC-C18和LC-NH2固相萃取柱净化、W aters ACQU ITYTMU PLC BEH-C18色谱柱(100mm×2.1mm,1.7μm)分离,在多反应监测模式下进行U PLC-MS/MS分析。7种性激素的方法检出限(S/N=3)为0.08～0.17μg/kg,定量限(S/N=10)为0.24～0.58μg/kg。考察了内标法和基质匹配外标法对7种性激素进行定量的回收率与精密度:添加水平为1,4μg/kg时,以内标法定量,7种性激素的平均回收率为76%～118%,相对标准偏差(RSD)为5.0%～11.3%;以基质匹配外标法定量,7种性激素的平均回收率为66%～94%,RSD为4.5%～10.7%。该结果表明两种方法均能够满足鱼制品中7种性激素的多残留检测要求。应用建立的方法对市售脱脂大黄鱼及烤鱼片进行检测,未发现7种目标违禁性激素。

超高效液相色谱-串联质谱法;性激素;多残留;鱼制品;ZnCl2去脂

Abstract:A rap id,specific and highly sensitive method for the determination of seven sexhormones(norgestrel,methyltestosterone,testosterone propionate,medroxy progesterone acetate,megestrol acetate,chlormadinone acetate,and nandrolone)residues in fish p roducts was developed using ultra performance liquid Chromatography-tandem mass spectrometry(U PLCMS/MS)with electrospray ionization(ESI)in positive m ode.The target com pounds were extracted with m ethanol after the enzyme hydrolysis of the fish products.ZnCl2was added to the extract solution to remove lip ids.Then target com pounds were purified by an LC-C18and an LCNH2solid phase extraction cartridges.The target com pounds were separated on a Waters ACQU ITYTMU PLC BEH-C18column(100mm×2.1mm,1.7μm)and detected qualitatively and quantitatively inmulti reaction monitoring(MRM)m ode.For the seven sexhormones,the limits of detection(LOD)of the method were from0.08to0.17μg/kg and the limits of quantification(LOQ)were in the range of0.24-0.58μg/kg.A t the spiked levels of1and4μg/kg,the average recoveries ranged from76%to118%with the relative standard deviations between 5.0%and11.3%for the seven sexhormones using internal standard method;and the average recoveries ranged from66%to94%with the relative standard deviations between4.5%and 10.7%using matrix m atched external standard method.The results show ed that both methods are able to meet the m ulti-residue detection of the seven sex hormone residues in fish products.The degreased large yellow croaker and roast fish fillet real samples from a local market were detected by the developed method,and the seven targets were not found.

Key words:ultra perform ance liquid Chromatography-electrosp ray tandem mass spectrometry(U PLC-MS/MS);sexhormones;multi-residues;fish p roducts;ZnCl2delipidation

食用动物饲养中促生长性激素的使用已经成为全球性的食品安全问题。研究表明,儿童性早熟、妇女乳腺癌和子宫癌发病率上升与动物性食品中性激素残留有关[1,2],因而欧盟在1996年[3]、我国在2002年[4]相继颁布法令,禁止合成性激素及具有促生长效用的物质作为促生剂在食用动物饲养中使用。……