超高效液相色谱法测定鸡组织中二硝托胺及其代谢产物残留量

Simultaneous Determination of Zoalene ant Its Metabolite in Chicken Tissues by Ultra Performance Liquid Chromatography

建立了超高效液相色谱同时测定鸡组织中二硝托胺及其代谢产物(3-Amino-5-nitro-o-toluamide,ANOT)的方法。样品用酸性磷酸盐缓冲液提取两次,离心后合并提取液,并调至pH 7.0,过HLB固相萃取小柱,用0.1%甲酸溶液-乙腈(80∶20,V/V)洗脱,洗脱液过滤膜后进行超高效液相色谱分析,二极管阵列检测器检测,外标法定量。二硝托胺和ANOT的检出限分别为25和55μg/kg。在添加浓度1500～9000μg/kg范围内,二硝托胺和ANOT的平均添加回收率为79.2%～88.1%,批内相对标准偏差(RSD)在2.7%～5.2%之间,批间RSD在4.1%～6.9%之间。峰面积与样品浓度在20～2000μg/L范围内呈良好的线性关系,二硝托胺和ANOT的线性回归系数均大于0.9998。

A method was developed for the simultaneous determination of zoalene and its metabolite（3-Amino-5-nitro-o-toluamide,ANOT） in chicken tissues by ultra performance liquid chromatog-raphy（UPLC）.Samples were extracted twice with acidic phosphate buffer solution and then the extracts were adjusted to pH 7.0 and purified through HLB cartridge after centrifugation.The residues were eluted from cartridge with 0.1% formic acid solution/acetonitrile（80 ∶ 20,V/V）.The samples were analyzed by UPLC with photodiode-array detector（PDA） and quantified with the external standard calibration curve method.The detection limits of ANOT and zoalene were 55.0 and 25.0 μg/kg,respectively.The mean recoveries of zoalene and ANOT in chicken tissues were 79.2%-88.1%,the intra-assay relative standard deviations were 2.7%-5.2% and the inter-assay relative standard deviations were 4.1%-6.9% at spiked levels of 1500-9000 μg/kg.There was a good linear correlation（the calibration coefficient is above 0.9998） between the peak areas and concentration of zoalene and ANOT in the range of 20 to 2000 μg/L.