高效液相色谱法测定猪肉中氨基比林、安替比林和安乃近代谢物残留量的研究

Study on Detection Method Aminopyrine and Antipyrine and Dipyrone Metabolite Residues in Pig Muscle by HPLC

建立了一种可同时检测猪肉中氨基比林、安替比林和安乃近代谢物4-甲氨基安替比林药物残留的高效液相色谱-紫外检测法。猪肉样品经乙腈提取,加入Na2SO4盐析去杂质,MCX柱净化。色谱柱为Phenomenex Luna C18(250 mm×4.6 mm,5μm),流动相为乙腈和水(20∶80),流速1.0 m L/min,检测波长265 nm。结果表明,氨基比林、安替比林和4-甲氨基安替比林在50~2000 ng/m L范围内呈良好的线性关系,R2均大于0.999。方法的最低定量限为20μg/kg。氨基比林、安替比林和4-甲氨基安替比林在20~400μg/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%。该方法具有简便快捷、灵敏度高等特点,适用于猪肉中氨基比林、安替比林和安乃近代谢物残留检测。

A method for determination of aminopyrine and antipyrine and dipyrone metabolite 4-methylaminoantipyrine in pig muscle with HPLC by UV detection was established. Aminopyrine and antipyrine and 4-methylaminoantipyrine were extracted from pig muscle with acetonitrile and spiked Na 2SO 4, dissolved with 2% formic acid after evaporation, then the solution was purified with MCX column. The chromatography column was Phenomenex Luna C18 column of 250 mm×4.6 mm, 5 μm. The mobile phase were acetonitrile and water, the flow rate was 1.0 mL/min and detection wavelength was 265 nm. A good linearity of the calibration curve was obtained with the range of 50～2000 ng/mL （ R 2〉0.999）,and the limit of quantification of aminopyrine and antipyrine and 4-methylaminoantipyrine was 20 μg/kg. The average recoveries of aminopyrine and antipyrine and 4-methylaminoantipyrine from spiked pig muscle at the three concentrations of 20,100,400 μg/kg ranged from 60% to 100%（ n =5） respectively. In day-between days batch variation coefficients were both less than 20%.The method is simple, rapid, sensitive and accurate, and can be applied for the determination of aminopyrine and antipyrine and dipyrone metabolite 4-methylaminoantipyrine in pig muscle.