超高效液相色谱串联质谱法测定畜产品中硝基呋喃类抗生素代谢物残留的研究

Determination of the Metabolites of Nitrofuran Antibiotics Residues in Animal Tissues by UPLC-MS/MS

建立和发展了畜产品4种硝基呋喃类抗生素代谢物残留的超高效液相色谱串联质谱法（UPLC—MS／MS）快速检测方法。样品中与蛋白结合的代谢物经0．125moL盐酸水解，2-硝基苯甲醛（2-NBA）37℃衍生16h。上清液调节pH=7．4，用乙酸乙酯提取，正己烷净化，流动相定容。采用电喷雾电离，正离子扫描，多反应检测模式（MRM）。外标法定量，在添加浓度为0．1～2．0μg／kg范围内，4种代谢物的平均回收率在67．3％～85．0％之间，10次测定结果的RSD平均值在2．0％～9．7％之间。最低检出限均达到0．15μg／kg，在0．10～20．00ng／mL的线性范围内，相关系数r均大于0．99，实现了样品检测的快速、高灵敏和高的选择性，适用于畜产品猪肉和猪肝中硝基呋喃类代谢物残留的检测。

A UPLC - MS/MS method was developed for the determination of the four metabolites of nitrofuran antibiotics in animal tissues. Protein - bound metabolites were released in HCL solution and derivafised with 2 - nitro-benzaldehyde overnight at 37 ℃. The derivatized solutions were adjusting pH to 7.4, then the analytes were extracted by ethyl acetate and cleaned up by n -hexane. Identification was achieved by electrospray ionization in positive mode ESI^＋ using multiple reaction monitoring （MRM）. The quantification was performed with external standards. The average rates of return were in the range of 67.3% ～85.0% with spiked levels of 0.1 ～ 2.0 μg/kg. The RSDs were 2.0%～9.7% （n = 10）. The limits of detection were 0.15 μg/kg for AMOZ, AOZ, SC and AHD. The linear ranges were from 0.10 to 20 ng/mL with correlation coefficients of 0.99. This method can be applied for the determination of metabolites of nitrofuran antibiotics in livestock products such as pork and pig liver because of its high speed, high sensitivity and high selectivity.