QuEChERS-高效液相色谱-紫外法测定动物性食品中氨基比林、安替比林和4-甲氨基安替比林的残留量

Determination of aminopyrine, antipyrine and 4-methylaminoantipyrine residues in animal derived food by QuEChERS-high performance liquid chromatography-violet wavelength detector

目的建立高效液相色谱-紫外法同时检测猪、牛、羊的肌肉、肝脏、肾脏中氨基比林、安替比林和安乃近代谢物4-甲氨基安替比林药物残留量的分析方法。方法样品加入4.0 g硫酸钠和1.0 g氯化钠盐析去杂质,并加入15 mL乙腈进行提取,离心后上清液加入到Agilent dSPE净化管中,涡旋混匀离心,再取上清液氮气吹干, 2%甲酸水溶液溶解后再过MCX柱净化。色谱柱为Phenomenex Luna C18 (250 mm×4.6 mm, 5μm),流动相为30%甲醇水溶液和乙腈,梯度洗脱,流速为1.0mL/min,检测波长265nm。结果氨基比林、安替比林和4-甲氨基安替比林在50～2000 ng/mL范围内具有良好的线性关系, r2均大于0.999。方法的最低定量限为50μg/kg。氨基比林、安替比林和4-甲氨基安替比林在50～200μg/kg的浓度添加水平上,其回收率在60%～100%,批内、批间的相对标准偏差小于20%。结论该方法具有简便快捷、灵敏度高等特点,适用于猪、牛、羊的肌肉、肝脏、肾脏中氨基比林、安替比林和安乃近代谢物残留检测。

Objective To establish a method for simultaneous determination of aminopyrine and antipyrine and dipyrone metabolite 4-methylaminoantipyrine in animal derived food by QuEChERS method coupled with high performance liquid chromatography-variable wavelength detector (HPLC-VWD). Methods Aminopyrine, antipyrine and 4-methylaminoantipyrine were extracted with 15 mL acetonitrile. Anhydrous sodium sulfate and anhydrous sodium chloride were used for the salting-out process, and the supernatant was added in Agilent dSPE column and mixed efficiently, centrifuged and dissolved with 2% formic acid after evaporation, and the solution was purified with MCX column. The chromatography column was Phenomenex Luna C 18 column (250 mm×4.6 mm, 5μm). The mobile phase was acetonitrile and 30% methanol-water with gradient elution, the flow rate was 1.0 mL/min and detection wavelength was 265 nm. Results Aminopyrine, Antipyrine and 4-methylaminoantipyrine had good linear relationships in the range of 50-2000 ng/mL (r 2 >0.999), and the limits of quantification was 50 μg/kg. The average recoveries of aminopyrine, antipyrine and 4-methylaminoantipyrine at 3 spiked levels (50, 100, 200 μg/kg) were 60%-100% (n=5). The relative standard deviation in batch and between batch were both less than 20%. Conclusion This method is simple, rapid, sensitive and accurate, which can be applied for the determination of aminopyrine, antipyrine and dipyrone metabolite 4-methylaminoantipyrine in animal derived food.