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

关键词:  高效液相色谱法  氨基比林  安替比林  安乃近  4-甲氨基安替比林  残留  猪肉

Key words:  HPLC  Aminopyrine  Antipyrine  Dipyrone  4-Methylaminoantipyrine  Residues  Pig muscle

摘要:

建立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 (250×4.6 mm，5 μm)，流动相为乙腈和水（20：80），流速为1.0 mL/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/mL呈良好的线性关系，R2均大于0.999。方法的最低定量限为20 μg/kg。氨基比林、安替比林和4-甲氨基安替比林在20~400 μg/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。

Abstract:

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 Na2SO4, dissolved with 2 % formic acid after evaporation, then the solution was purified with MCX column. The chromatography column was Phenomenex Luna C18 column of 250×4.6mm, 5μm. The mobile phase were acetonitrile and water, the flow rate was 1.0mL/min and detection wavelengths was 265 nm. A good linearity of the calibration curve was obtained with the range of 50-2000 ng/mL （R2>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 60-100%(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.