猪可食性组织中氟尼辛残留量的UPLC-MS/MS检测方法的建立

Development of an ultra-performance liquid chromatography-tandem mass spectrometry method for the determination of flunixin in edible swine tissues

[目的]建立一种可靠、快速、灵敏的超高效液相色谱-串联质谱法(UPLC-MS/MS)用于猪可食性组织中氟尼辛残留量的检测。[方法]猪可食性组织(肌肉、皮脂、肝脏、肾脏)经绞碎、均质后,用乙酸乙酯-乙腈(体积比1∶1)提取,MCX固相萃取柱净化,Acquity UPLC BEH C_(18)色谱柱梯度洗脱,在电喷雾正离子(ESI+)下选用多反应监测(MRM)模式进行待测物的分析,外标法定量。[结果]该方法的检测限(LOD)与定量限(LOQ)分别为0.5μg·kg-1和1.0μg·kg^(-1);待测物氟尼辛含量在0.5~500μg·kg^(-1)范围内,峰面积与药含量呈良好的线性关系(R2>0.999);氟尼辛在猪肌肉、皮脂、肝脏、肾脏组织中的添加回收率为74.8%~90.3%,批内精密度及批间精密度均不大于16.5%。[结论]建立了猪超高效液相色谱-串联质谱检测方法,适用于猪可食性组织中氟尼辛残留量的测定。

[ Objectives] To establish a reliable, rapid and sensitive method for determination of flunixin residues in swine tissues （muscle, skin ＋ fat, liver, kidney ）, ultra-performance liquid chromatography-tandem mass spectrometry （ UPLC-MS/MS ） was developed and optimized. [ Methods] The samples were extracted with acetonitrile and ethyl acetate （volume ratio 1：1 ）, and then purified via an Oasis MCX solid-phase extraction cartridge. Chromatographic separation was performed on an Acquity UPLC BEH C18 column. The analysis was carried out on UPLC-ESI-MS/MS working with multiple reaction monitoring mode. External standard was used for quantitation of target drug. [ Results ] The analyte has a good linearity at the range of 0.5-500 μg·kg^-1, with related coeffi- cient of R2〉0.999. Recoveries of spiked samples ranged from 74.8% to 90.3% with relative standard deviation （RSD）less than 16.5%. The limits of detection（LOD） and quantification （LOQ） were 0.5 and 1.0 μg·kg^-1 , respectively. [ Conclusions ] The results show that the pretreatment and purification process established by this study are easy to operate. The method has good stability, reproducibility, sensitivity, and great accuracy.