高效液相色谱-串联质谱法同时测定饲料中乙氧基喹啉及其伴生物质

Simultaneous Determination of Ethoxyquin and Its Associated Substances in Feeds by High Performance Liquid Chromatography-Tandem Mass Spectrometry

本研究旨在建立一种应用高效液相色谱-串联质谱(HPLC-MS/MS)技术同时测定饲料中乙氧基喹啉(EQ)、对氨基苯乙醚(PD)、非那西丁(PC)、乙氧基喹醌亚胺(EQI)和乙氧基喹啉二聚体(EQDM)含量的方法。饲料样品用乙腈提取,经BONDESIL-SI多重杂质吸附材料净化,采用HPLC-MS/MS测定,以0.1%甲酸水溶液-0.1%甲酸乙腈溶液作为流动相,梯度洗脱,Waters BEH C_(18)色谱柱分离。结果表明:通过饲料基质匹配绘制标准曲线,EQ、PD、PC、EQI和EQDM在5~500μg/L范围内线性关系良好[决定系数(R^(2))>0.9950]。方法中EQ、PD和PC的检出限和定量限分别为3.0和10.0μg/kg,EQI和EQDM的检出限和定量限分别为1.5和5.0μg/kg。EQ、PD、PC、EQI和EQDM在3个添加水平(20、50、100μg/kg)的平均回收率为73.77%~98.77%,批内相对标准偏差(RSD,n=6)<10%,批间变异系数<12%。该方法前处理简单,操作方便,适用于饲料中EQ及其伴生物质的同步测定。

The aim of this study was to establish a method for simultaneous determination of ethoxyquin(EQ),p-phenetidine(PD),phenacetin(PC),ethoxyquin quinone imine(EQI)and ethoxyquin dimer(EQDM)in feeds by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).The feed samples were extracted with acetonitrile,purified by BONDESIL-SI multiple impurity adsorption material,determined by HPLC-MS/MS.The samples were separated with 0.1% formic acid aqueous solution and 0.1% formic acid acetonitrile solution as mobile phase by gradient elution,and separated by Waters BEH C_(18) column.The results showed that the standard curves of EQ,PD,PC,EQI and EQDM were well linear in the range of 5 to 500μg/L[coefficient of determination(R^(2))>0.9950].The limits of detection and quantification for EQ,PD and PC were 3.0 and 10.0μg/kg,and those for EQI and EQDM were 1.5 and 5.0μg/kg,respectively.The average recovery rates of EQ,PD,PC,EQI and EQDM at the three supplemental levels(20,50 and 100μg/kg)ranged from 73.77% to 98.77%,the relative standard deviation(RSD,n=6)within the lot was<10%,and the coefficient of variation between the batches was<12%.The method is simple and easy to operate,and is suitable for simultaneous determination of EQ and its associated biomass in feeds.