高效液相色谱-串联质谱法检测牛组织中双氯芬酸钠残留的研究

Detection of diclofenac sodium residue in bovine tissues using high performance liquid chromatography-tandem mass spectrometry

为了测定牛肌肉、肝脏、肾脏组织中双氯芬酸钠的残留量,研究建立了一种用于该检测的高效液相色谱-串联质谱(LC-MS/MS)方法。试验将样品中加入乙腈提取液重复提取2次,超声波辅助提取,提取液中加入C_(18)、PSA混合吸附剂进行净化。色谱柱采用C_(18)反相柱对药物进行分离,洗脱液采用乙腈-0.1%甲酸水进行等度洗脱,配备电喷雾离子源,药物在正离子模式下电离,多反应监测模式(MRM)分段扫描测定检测结果,并采用内标法定量分析。结果表明,双氯芬酸在0.5~100μg/L浓度范围内有良好的线性关系,相关系数大于0.99;在牛的肌肉、肝脏和肾脏组织中检测限(LOD,S/N=3)为0.5μg/kg,定量限(LOQ,S/N=10)为1.0μg/kg。在3个浓度添加水平下,平均回收率为97.7%~114%,日内变异系数0.5%~4.7%,日间变异系数为0.9%~7.5%。该方法操作简便、结果准确,满足兽药残留检测方法规定要求,运用于快速准确测定牛肌肉、肝脏、肾脏等组织中双氯芬酸钠的残留。

In order to determine the content of diclofenac sodium residue, a method with high performance liquid chromatography-tandem mass spectrometry to detect drug residues in the tissues of bovine muscle, liver and kidney was established. Samples were extracted by acetonitrile twice. C_(18)and PSA mixed adsorbent was added for purification. The C_(18)reversed-phase column was used to separate the drugs, and acetonitrile-0.1% formic acid water was used as a mobile phase with equal elution. The scanning was monitored in a multi-response monitoring mode(MRM), applying a positive ions electrospray ion source mode simultaneously. And the internal standard method was used for quantitative analysis. The results showed that the linear relationship of diclofenac in a 0.5-100 μg/L concentration range was good, and the correlation coefficient was over 0.99. The detection limit(LOD, S/N= 3) of this method was 0.5 μg/kg and the limit of quantitation(LOQ, S/N= 10) was 1.0 μg/kg. At the level of 3 concentrations, the average recovery rate was 97.7%-114%, the intra-batch variation coefficient was 0.5%-4.7%, and the inter-batch coefficient variation was 0.9%-7.5%. The present method met the requirements for veterinary drug residue trace detection. The method was time-efficient, easy and accurate, and was suitable for rapid screening of diclofenac sodium residue in bovine tissue.