应用HPLC-MS/MS对鸡组织中三种大环内酯类抗生素残留的同时检测

Simultaneous determination of three macrolide antibiotics residues in chicken by HPLC-MS/MS

为建立鸡体内阿奇霉素(AZI)、替米考星(TILM)、泰乐菌素(TYL)三种大环内酯类抗生素残留的高效液相色谱-串联质谱检测方法,样品经乙腈提取,MCX柱净化,Luna C8色谱柱分离,以1mL/L的甲酸-乙腈和1mL/L甲酸-水作为流动相进行梯度洗脱以及电喷雾正离子多反应模式检测。结果显示,3种抗生素在1～100ng/mL范围内呈现良好的线性关系,相关系数R2均大于0.998,AZI、TILM、TYL的检测限分别为0.1、0.2、0.3μg/kg,定量限依次为0.3、0.7、1.0μg/kg。以0.5、5、20μg/kg共3个水平进行添加回收试验,3种药物的回收率为83.5%～108.0%,日内相对标准偏差为4.3%～15.2%,日间相对标准偏差为4.8%～11.3%。结果表明,该方法简便快速,灵敏度高,适用于鸡体内三种大环内酯类残留的同时检测。

In order to establish the high performance liquid chromatography tandem mass spectrometry（HPLC-MS/MS） for the determination of there macrolide antibiotics（tylosin,tilmicosin and azithromycin） in chicken,samples were extracted with acetonitrile and cleaned-up by MCX cartridge.The three antibiotics were separated with luna C8 column by a gradient elution program with 1 mL/L formic acid-acetonitrile and 1 mL/L formic acid-water as the mobile phase.The identification of three drugs was carried out by positive electrospray ionization（ESI＋） in multiple reaction monitoring（MRM） mode.In result,the calibration curves showed good linearity for 3 antibiotics in the range from 1 ng/mL to 100 ng/mL,with correlation coefficient greater than 0.998.The limits of detection（LOD） were 0.1,0.2 and 0.3 μg/kg,respectively and the limits of quantification（LOQ） were 0.3,0.7,1.0 μg/kg for AZI,TILM and TYL,respectively.The average recoveries varied from 83.5% to 108.0% at three spiked levels of 0.5,5 and 20 μg/kg.The intra-RSDS were in the range from 4.3% to 15.2% and inter-RSDS were from 4.8% to 11.3%.The result showed that the method is simple,rapid,sensitive and suitable for the simultaneous determination of 3 macrolide antibiotics in chicken.