Simultaneous determination of florfenicol and florfenicol amine in eggs and chicken by ultraperformance liquid chromatography-tandem mass spectrometry with automatic solid phase extraction
全自动固相萃取-超高效液相色谱-串联质谱法同时测定鸡蛋和鸡肉中的氟苯尼考和氟苯尼考胺残留

摘要

目的 建立全自动固相萃取-超高效液相色谱-串联质谱法同时测定鸡蛋和鸡肉中的氟苯尼考与氟苯尼考胺残留的检测方法。方法 样品以水稀释后，以氨水-乙酸乙酯（3:100）溶液提取，经C18固相萃取柱净化，用100%甲醇洗脱浓缩后，采用ACQUITY UPLC® BEN C18色谱柱 （2.1mm*100mm，1.7μm）分离，以0.02% 氨水-乙腈为流动相，用正负离子多反应监测模式，内标法定量。结果 目标化合物得到有效分离，方法的线性关系良好（r2＞0.999），氟苯尼考的检出限0.0057μg/kg，定量限0.0019μg/kg，氟苯尼考胺的检出限0.022μg/kg，定量限0.0074μg/kg。高中低三种添加水平的加标回收试验：鸡蛋的加标回收率为91.9%～108.6%，相对标准偏差为7.1%～14.2%，鸡肉的加标回收率为94.2%～117.2%，相对标准偏差为1.4%～7.6%。应用该方法对广东省的鸡蛋和鸡肉样品各60份进行检测，结果在14份鸡肉中有检出氟苯尼考或氟苯尼考胺，其中在1份鸡肉样品中同时检出氟苯尼考和氟苯尼考胺，结果分别为320μg/kg和120μg/kg，均超出限量值（100μg/kg）。而鸡蛋样品均未检出。结论 本方法准确、灵敏，为鸡肉和鸡蛋中氟苯尼考、氟苯尼考胺残留同时检测和监测提供更可靠的分析手段。

Abstract

Objective To establish a method for the simultaneous determination of florfenicol and florfenicol amine residues in eggs and chicken by UPLC-MS/MS with automatic solid phase extraction.
Methods The samples were diluted with water, extracted with ammonia-ethyl acetate (3:100), and the resulting extracts were clean-up on C18 solid phase extraction column,and then eluted with 100% methanol. The target components were separated on a ACQUITY UPLC ® Ben C18 column (2.1mm * 100mm, 1.7μm) , with 0.02% ammonia water and acetonitrile as mobile phase. Multiple reaction monitoring in positive and negative mode were used，and it quantified by the internal standard calibration curves.
Results The target components could be well separated，with coefficients of determination higher than 0.999．The limits of detection and limits of quantification of florfenicol were 0.0057 and 0.0019μg/kg，The limits of detection and limits of quantification of florfenicol amine were 0.022 and 0.0074μg/kg．The recovery tests at high，medium，and low spiked levels: the recoveries were 91.9%-108.6%, and relative standard deviation (RSDs) were7.1%-14.2% in eggs; the recoveries were 94.2%-117.2%，and RSDs were 1.4%-7.6% in chicken. The method was used to detect 60 eggs and 60 chicken samples from Guangdong Province, and the result showed that florfenicol or florfenicol amine were detected in 14 chicken samples，they were detected simultaneously in one chicken sample, and the results were 320 μg/kg and 120μg/kg，which exceeded the limit value of 100μg/kg. It was not detected in egg samples.
Conclusion   The method was accurate and sensitive, and provides a more reliable analytical method for the simultaneous detection and monitoring of florfenicol and florfenicol amine residues in chicken and eggs.