基于实验室能力验证需求国标法检测鸡肉和鸡蛋中氟喹诺酮类药物残留量的优化

Optimization of the National Standard Method for Determining the Fluoroquinolones Residues in Chicken and Eggs Based on Laboratory Proficiency Testing Requirements

基于实验室能力验证检测工作的特点,对农业部1025号公告-14-2008和农业部781号公告-6-2006两个国标法的液相色谱检测方法进行优化,结合加热沉淀法,快速测定鸡肉、鸡蛋样品中氟喹诺酮类药物的残留量,为依照国标法检测步骤完成能力验证实验的结果提供质量验证参比。鸡肉或鸡蛋考核样品按国标方法提取获得备用液(其中鸡蛋备用液定容至6 mL),精密移取待固相萃取(SPE)净化的备用液1 mL,加入乙腈0.2 mL,涡旋,55℃水浴加热20 min快速沉淀蛋白质,离心后取上清液进高效液相色谱仪快速测定样品中的药物残留量。快速加热沉淀法应用于鸡肉、鸡蛋样品时,添加回收率分别为81.08%~103.09%和77.60%~102.94%。剩余备用液依照国标法继续净化,并对净化操作稍作优化:鸡蛋备用液只取3 mL进行SPE净化;鸡肉备用液先冷冻,解冻后高速离心,移取上清液进行SPE净化。优化方案操作简单,能快速测得样品中氟喹诺酮类药物的浓度,提高了回收率和重复性,既可为能力验证提供结果质量验证的参考,亦可用于日常的批量快速检测。

Based on the characteristics of the laboratory proficiency testing and verification,the liquid chromatography analysis in the two national standard methods of the No.1025-14-2008 announcement and No.781-6-2006 announcement from the Ministry of Agriculture of the People’s Republic of China are optimized.In combination with the thermal precipitation process,rapid determination of the fluoroquinolone residues in chicken and egg samples could be accomplished,which provides a quality verification reference for the results of the proficiency testing and verification following the procedure of the national standard method.The test samples of chicken or eggs were extracted according to the national standard method to obtain the standby liquid(with the volume of the egg standby liquid was fixed to 6 mL).1 mL of the standby liquid was precisely withdrawn for solid phase extraction(SPE)purification,then,the protein was rapidly precipitated by adding 0.2 mL of acetonitrile,vortexing,and heating for 20 min in a water bath at 55℃.After centrifugation,the supernatant was collected for rapid determination of the drug residue by high performance liquid chromatography.When the rapid heat precipitation method was applied to chicken and egg samples,the recoveries were 81.08%~103.09%and 77.60%~102.94%,respectively.The remaining standby liquid was further purified in accordance with the national standard method,and the purification processes were slightly optimized as follows:only 3 mL of the standby liquid of egg was taken for SPE purification;whilst the standby liquid of chicken was firstly frozen,then thawed,and centrifuged at a high speed.The supernatant was collected for SPE purification.Overall,the optimized procedure is simple with improved recovery and repeatability,and can not only provide a quality verification reference for the results of proficiency testing and verification,but also allow daily rapid batch detection.