鸡蛋中硝基呋喃类代谢物残留检测UPLC-MS/MS法研究

Determination of Nitrofurans Metabolites Residue in Egg by UPLC-MS/MS

关键词:  鸡蛋  硝基呋喃类代谢物  超高效液相色谱-串联质谱

Key words:  Nitrofurans metabolites  egg  UPLC-MS/MS

摘要:

建立了鸡蛋中硝基呋喃类代谢物（AOZ、AMOZ、AHD和SEM）残留检测的超高效液相色谱-串联质谱方法（UPLC-MS/MS）。样品在酸性条件下衍生化后，弱碱性条件下用乙酸乙酯提取后，用正己烷去除脂肪，高速离心去除蛋白质等杂质，上机测试。液相色谱条件：色谱柱为BEH C18（50×2.1mm，1.7μm），流动相为甲醇+10mmol/L乙酸铵水溶液，梯度洗脱，流速为0.3 mL/min，柱温为30 ℃，进样量为10 μL。质谱条件：电喷雾离子源（ESI+），多反应监测（MRM）方式进行采集。结果表明：硝基呋喃类代谢物在0.5~20 ng/mL浓度范围内均呈现良好的线性关系，相关系数（R2）均大于0.990；鸡蛋中四种代谢物的检测限均为0.25ng/g，定量限均为0.5ng/g。在0.5、1和5 ng/g三个添加浓度平均回收率为79.1％~109.4％，批内和批间RSD均小于20%。该方法具有简便快捷、灵敏度高、定性准确等特点。

Abstract:

A UPLC-MS/MS method was established for the determination of Nitrofurans metabolites residue in chicken egg. The samples were extracted by ethyl acetate in alkaline conditon, then the fat was removed by hexane, and the protein was deposited by high speed centrifugation. LC conditions were as follows: the chromatography column is BEH C18 column of 50×2.1 mm, 1.7 μm；mobile phase is methanol and water（10mmol/L ammonium acetate）; column temperature is 30 ℃; flow rate is 0.3 mL/min; injection volume is 10 μL. Mass spectrometry conditions were ESI and MRM mode. The calibration curve of Nitrofurans metabolites were good linear from 0.5 to 20 ng/mL with the correlation coefficient R2 over 0.990, respectively. The limit of detection of the method was 0.25ng/g, and the limit of quantification was 0.5ng/g. The average recoveries of Nitrofurans metabolites from spiked egg at three concentrations of 0.5, 1 and 1.5ng/g were 79.1％~109.4％, and intra- and inter-batch RSD were <20%.