UPLC-MS/MS法测定禽蛋中氟苯尼考及氟苯尼考胺

建立一种超高效液相色谱-串联质谱法同时测定禽蛋中氟苯尼考(FF)和氟苯尼考胺(FFA)含量的方法。样品经2%氨水-乙酸乙酯提取,PRiME HLB固相萃取(SPE)柱净化。采用Hypersil GOLD C18色谱柱(100 mm×2.1mm,1.9μm),以乙腈(B1)-0.1%甲酸水(A2)为流动相进行梯度洗脱。氟苯尼考和氘代氟苯尼考采用电喷雾负离子源,氟苯尼考胺采用电喷雾正离子源,多反应监测模式,以氘代氟苯尼考(氟苯尼考-d3)为氟苯尼考进行内标定量,氟苯尼考胺外标法定量。结果表明,氟苯尼考和氟苯尼考胺在0.2~10μg/L范围内均呈良好线性关系(r≥0.9978)。加标浓度分别为0.1,0.2和1μg/kg及0.5,1.0和5.0μg/kg,回收率分别在91.0%~100.9%和73.7%~84.3%之间,相对标准偏差均<3.52%(n=6)。氟苯尼考的检出限和定量限分别为0.05μg/kg和0.1μg/kg,氟苯尼考胺的检出限和定量限分别为0.25μg/kg和0.5μg/kg。该方法准确、灵敏、快速,能有效并同时检测禽蛋中氟苯尼考和氟苯尼考胺含量。

Simultaneous Determination of Florfenicol and Florfenicol Amine in Eggs by UPLC-MS/MS

A reliable ultra performance liquid chromatography-mass spectrometry/mass spectrometry(UPLC-MS/MS)method was developed to determine florfenicol(FF)and its metabolite florfenicol amine(FFA)in eggs.Analytes were extracted with 2%ammonium hydroxide-ethyl acetate,followed by Oasis PRiME HLB Extraction Cartridge cleanup.These compounds were separated by a Hypersil GOLD C18(100 mm×2.1 mm,1.9μm)column using acetonitrile(B1)-0.1%formic acid water(A2)as the gradient elution of mobile phase.FF and FF-d3 were analyzed with electrospray ionization in the negative ion mode and FFA was analyzed in the positive ion mode by multiple reaction monitoring(MRM).FF and FF-d3 were quantified by internal calibration while FFA was quantified by external method.Calibration curves were linear in the range of 0.2 to 10μg/L,with coefficients of determination higher than 0.9978.Good recoveries were obtained(91.0%-100.9%and 73.7%-84.3%),with relative standard deviations of<3.52%for the two analytes.The limits of detection and limits of quantification of FF were estimated to be 0.05μg/kg and 0.1μg/kg,and the two limits of FFA were 0.25μg/kg and 0.5μg/kg,respectively in eggs.This method was effective for simultaneous determination of FF and FFA in eggs.