超高效液相色谱-串联质谱法同时测定动物组织中氯霉素、甲砜霉素、氟苯尼考和其代谢产物氟苯尼考胺残留量

Determination of Chloramphenicol,Thiamphenicol,Florfenicol and Its Metabolite Florfenicol Amine Residues in Animal Tissues by UHPLC-MS/MS

目的:建立超高效液相色谱-串联质谱法同时测定猪肉、鸡肉和鱼肉中氯霉素、甲砜霉素、氟苯尼考及其代谢物氟苯尼考胺的分析方法。方法:样品经碱化乙酸乙酯提取,正己烷去除油脂,通过MCX固相萃取柱净化,经Waters BEH C_(18)柱(100 mm×2.1 mm,1.7μm)分离,以0.1%氨水溶液-乙腈为流动相梯度洗脱,电喷雾正/负离子切换,多反应模式监测,同位素内标法定量。结果:氯霉素、甲砜霉素和氟苯尼考在0.5~100.0μg·L^(-1)具有良好的线性关系,相关系数(R^(2))均大于0.999,检出限均为0.1μg·kg^(-1),方法回收率为83%~112%,相对标准偏差小于10%;氟苯尼考胺在1~100μg·L^(-1)具有良好的线性关系,相关系数(R^(2))大于0.999,检出限为0.5μg·kg^(-1),方法回收率为81%~110%,相对标准偏差小于10%。结论:本方法重现性好、灵敏、准确,适用于日常大批量猪肉、鸡肉和鱼肉样品中氯霉素类药物及其代谢物残留的定性、定量分析。

Objective:To establish a method for the determination of chloramphenicol,thiamphenicol,florfenicol and its metabolite florfenicol amine residues in pork,chicken and fish by ultra-high performance liquid chromatography-tandem mass spectrometry.Method:The sample was extracted with alkalized ethyl acetate,oil was removed with n-hexane,purified by MCX solid phase extraction column.An Waters BEH C_(18) column(100 mm×2.1 mm,1.7μm)was used for the separation by gradient elution program using 0.1%ammonina-acetonitrile as mobile phase gradient.The samples were simultaneous determined by mass spectrometer with electrospray ion source in the positive and negative ion mode and monitored by the multiple reaction monitoring,and quantified by isotope internal standard method.Result:The chloramphenicol,thiamphenicol and florfenicol showed good linearity in the range of 0.5~100.0μg·L^(-1) with the quantification limit of 0.1μg·kg^(-1),The correlation coefficients(R^(2))were greater than 0.999,and the recovery rates were between 83%and 112%with the RSD less than 10%.The florfenicol amine showed good linearity in the range of 1~100μg·L^(-1) with the quantification limit of 0.5μg·kg^(-1).The correlation coefficient(R^(2))was greater than 0.999.The recovery rate was between 81%and 110%,with the RSD less than 10%.Conclusion:The method is reproducible,sensitive and accurate,and could be applied for the qualitative and quantitative analysis of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine residues antibiotics in pork,chicken and fish.