通过式固相萃取净化-超高效液相色谱-串联质谱法快速测定禽肉中利巴韦林的残留量

Rapid Determination of Ribavirin Residue in Poultry Meat through Purification by Solid-phase Extraction and Analysis by Ultra-performance Liquid Chromatography-tandem Mass Spectrometry

建立了通过式固相萃取净化-超高效液相色谱-串联质谱法快速测定禽肉中利巴韦林残留量的方法。样品在37℃经酸性磷酸酯酶酶解,乙腈提取,EMR-Lipid固相萃取柱净化,以乙腈和0.1%甲酸水(含5 mmol/L甲酸铵)为流动相,经过BEH Z-HILIC色谱柱(100 mm×2.1 mm,1.7μm)分离,经串联质谱仪,在电喷雾正离子模式下以多反应监测(MRM)模式测定,同位素内标法定量。对前处理条件、液相条件和质谱参数进行了系统的考察。该方法在0.05~5μg/L质量浓度范围内线性关系良好,定量限(LOQ)为2.0μg/kg。在2.0、4.0、20μg/kg添加水平下,回收率为94.5%~112.0%,相对标准偏差(n=6)为1.4%~3.2%。该方法相比标准方法前处理简单、快速,适用于禽肉中利巴韦林残留的批量分析。

A rapid analysis method was established for determining the ribavirin residue in poultry meat through purification by solid-phase extraction and analysis by ultra-performance liquid chromatography-tandem mass spectrometry.After enzymatic hydrolysis with acid phosphatase at 37℃,the samples were extracted with acetonitrile and purified with an EMR-Lipid solid phase extraction column to remove liposoluble substances.The purified sample solution was subjected to separation using a BEH Z-HILIC column(100 mm×2.1 mm,1.7μm)and a mobile phase consisting of acetonitrile and 0.1%aqueous formic acid solution(containing 5 mmol/L ammonium formate)for analysis by tandem mass spectrometer in positive electrospray ionization mode with multiple reaction monitoring(MRM).An isotope-labeled internal standard method was used for quantitative analysis.The pretreat conditions,HPLC conditions and mass spectrometry parameters were systematically investigated.This method showed good linearity in the range of 0.05~5μg/L with the limit of quantification(LOQ)being 2.0μg/kg.When the spiking levels were 2.0,4.0 and 20.0μg/kg,the recoveries of ribavirin were in the range of 94.5%~112.0%with the relative standard deviations(n=6)being 1.4%~3.2%.Compared with the standard method,this method is simple and rapid,thus suitable for batch analysis of ribavirin residue in poultry meat.