超高效液相色谱-串联质谱法测定羊奶中5种阿维菌素类药物残留的研究

Study on the determination of five avermectin residues in goat milk by ultra-high performance liquid chromatography-tandem masss pectrometry

为了建立同时测定羊奶中5种阿维菌素类药物(avermectins, AVMs),包括阿维菌素(AVM)、伊维菌素(IVM)、多拉菌素(DOR)、埃玛菌素(EMM)、埃普利诺菌素(EPR)残留量的超高效液相色谱-串联质谱(UPLC-MS/MS)法,试验采用乙腈提取羊奶样品中的AVMs,提取液用C_(18)固相萃取柱净化后经C_(18)色谱柱分离,采用电喷雾离子源正离子(ESI+)在多反应离子监测(MRM)模式下同时测定5种AVMs,并用基质标准曲线回归方程定量,考察所建立方法的准确度和精密度,评价5种AVMs在6批不同来源羊奶中的基质效应,同时应用建立的方法检测10批鲜羊奶样品中5种AVMs。结果表明:IVM和EMM在0.5~50μg/L范围内以及AVM、DOM和EPR在2~200μg/L范围内线性关系良好,相关系数均达到0.999 6以上;方法的检出限为0.2~1.0μg/kg,定量下限为0.5~2.0μg/kg。羊奶基质中IVM和EMM在0.5,2,5μg/kg加标水平以及AVM、DOM和EPR在2,8,20μg/kg加标水平下,5种AVMs的回收率在73.5%~108.0%范围内,批内和批间相对标准偏差均小于15.0%。5种AVMs的基质效应均控制在85.0%~115.0%之间,基质效应差异不明显。试验建立的超高效液相-串联质谱法对10批鲜羊奶样品的检测结果与秦皇岛出入境检验检疫局的检测结果一致,仅从1份样品中检出IVM残留量为13.4μg/kg,其他样品均未检出AVMs残留。说明试验建立的方法简单、稳定、可靠,可用于羊奶中AVMs残留的检测。

In order to establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) for the simultaneous determination of the residues of five avermectins(AVMs) in goat milk, including avermectin(AVM),ivermectin(IVM),doramectin(DOR),emamectin(EMM) and eprinomectin(EPR),the experiment used acetonitrile for the extraction of AVMs from goat milk. The extracted solution was purified by C_(18) SPE column and separated by C_(18) chromatographic column. Five AVMs were determined simultaneously by positive ions of electrospray ion source(ESI+) under multi-reactive ion monitoring(MRM) mode. Matrix standard curve regression equation was used for the quantification;the accuracy and precision of the established method were investigated. The matrix effect of 5 AVMs in 6 batches of goat milk from different sources was evaluated;and the established method was used for the determination of five AVMs in 10 batches of fresh goat milk samples. The results showed that IVM and EMM within the range of 0.5-50 μg/L and AVM,DOM and EPR within the range of 2-200 μg/L,had good linear relationship;and the correlation coefficients were≥0.999 6. The limits of detection were 0.2-1.0 μg/kg, and the limits of quantification were 0.5-2.0 μg/kg. At 0.5,2,5 μg/kg spiked levels of IVM and EMM,or at 2,8,20 μg/kg spiked levels of AVM,DOM and EPR,the recoveries of five AVMs were in the range of 73.5%-108.0%,with intra-batch and inter-batch relative standard deviations less than 15.0%. The matrix effect of the five AVMs was controlled in the range of 85.0%-115.0%,and the matrix effect was not significant. The results of ten batches of goat milk samples detected by UPLC-MS/MS were consistent with those detected by Qinhuangdao Entry-Exit Inspection and Quarantine Bureau. Only one sample was detected with IVM residue of 13.4 μg/kg, and no AVMS residue was detected in other samples. This method was simple, stable and reliable, and could meet the needs of detection and confirmation of AVMS residues in goat milk.