超高效液相色谱-四极杆飞行时间质谱法测定饲料中9种雄性激素类药物

建立了超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF-MS)分析动物饲料中违禁添加的去氢睾酮(BOL)、表睾酮(epiAn)、氟甲睾酮(FT)、去氢甲睾酮(美雄酮)(MD)、甲睾酮(MT)、丙酸诺龙(NP)、诺龙(N)、丙酸睾酮(TP)和睾酮(T)9种药物的分析方法。样品经乙酸乙酯振荡提取,再用基质固相萃取方法净化处理后,采用UPLC-Q-TOF-MS分析检测。在电喷雾正离子模式和飞行时间模式下,输入各化合物的精确分子离子质量数得到相应的提取离子色谱图,以色谱峰面积进行定量分析。通过碰撞诱导解离模式(CID)得到各化合物碎片离子的精确质荷比,进一步对各化合物进行定性分析。各化合物的质量精确度均小于5×10-6,9种化合物在0～1000μg/L范围内均呈良好的线性关系,线性系数均大于0.99。除诺龙和去氢甲睾酮外,本方法对各药物的检出限(LOD)均低于6μg/L;去氢睾酮、表睾酮、氟甲睾酮、去氢甲睾酮(美雄酮)、甲睾酮、丙酸诺龙、诺龙、丙酸睾酮和睾酮的定量限(LOQ)分别为16,10,20,43,20,12,15,10g和16μg/kg。3个添加水平(LOQ,2LOQ,4LOQ)的回收实验表明,化合物的回收率在70.0%～99.7%范围内,相对标准偏差(RSD)均小于10%。本方法的定性准确度明显高于文献报道的方法,可用于饲料中的禁用雄性激素药物的测定。

Simultaneous Determination of Nine Androgen Drugs in Feeds by Ultra-Performance Liquid Chromatography-QuadrupoleTime-of-Flight Mass Spectrometry

An ultra-performance liquid chromatography(UPLC) quadrupole-time-of-flight mass spectrometric(Q-TOF-MS) method was developed for determination of 9 androgen drugs in feeds.After homogenization,the samples were extracted with ethyl acetate,and then the extracts were directly purified with matrix solid phase extraction.Identification and detection were achieved in the positive electrospray ionization(ESI) mode using Q-TOF-MS.The extracted ion chromatograms of each compound were obtained by entering the accurate molecular mass.The samples were quantified with the chromatographic peak area of each compound.The potential of UPLC-Q-TOF MS for confirmatory analysis was shown by determining the accurate mass of all compounds and fragment ions upon collision-induced-dissociation(CID) at different energies.The extra mass measurements errors for all androgen drugs were found to be within 5 ×10-6.For these drugs,the calibration curves were linear between 0 to 1000 μg/L,and the correlation coefficient was more than 0.99.The recoveries for all compounds in feeds were 70.0%-99.7% at the fortified levels of 1LOQ,2LOQ and 4LOQ with the limit of quantitation(LOQ) of 16,10,20,43,20,12,15,10 and 16 μg/kg.The relative standard deviations(RSD) for all compounds were less than 10%.