超高效液相色谱-串联质谱法快速测定鱼和虾中多类禁、限用兽药残留

Rapid determination of multiple prohibited and restricted veterinary drug residues in fish and shrimp by ultra high performance liquid chromatography-tandem mass spectrometry

建立了鱼和虾中5类(磺胺类、喹诺酮类、氯霉素类、硝基呋喃类代谢物、三苯甲烷类)28种禁、限用兽药的同时提取净化-超高效液相色谱-串联质谱(UHPLC-MS/MS)快速检测的方法。对样品前处理及色谱条件进行优化,以盐酸水解样品,待硝基呋喃类代谢物与2-硝基苯甲醛衍生化后,5类残留药物经乙腈提取,提取液经氯化钠脱水、盐析,乙腈层经正己烷净化后浓缩至近干,残留物用甲醇定容至1.0 mL,加50 mg C18、30 mg N-丙基乙二胺(PSA)、60 mg氨基键合硅胶填料(NH2)吸附剂分散固相萃取净化,最后经ZORBAX C18色谱柱(50 mm×2.1 mm,1.8μm)分离,在正、负离子多反应监测模式下采集,同位素内标法定量。所建立的方法确保28种兽药在各自范围内线性关系良好,相关系数(r)>0.99,方法的检出限和定量限分别为0.1~0.8μg/kg和0.3~2.7μg/kg;在1、5、20倍定量限添加水平下的平均回收率为70.0%~120%,相对标准偏差为0.9%~10.0%。方法简便、快速、准确、灵敏,适用于鱼和虾中多类禁、限用药物的快速测定。

A method was established for the rapid determination of 28 prohibited and restricted veterinary drugs of five classes(sulfonamides,quinolones,chloramphenicols,nitrofurans metabolites and triphenylmethane)in fish and shrimp by ultra high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)coupled with simultaneous extraction and purification.After optimizing the sample pretreatment methods and chromatographic conditions,the samples were hydrolyzed with hydrochloric acid.The five kinds of residual drugs were simultaneously extracted with acetonitrile after nitrofuran metabolites derivatized with 2-nitrobenzaldehyde.The extraction solutions were dehydrated with sodium chloride and salted out.The acetonitrile layer was purified with n-hexane and concentrated to near dryness.The residues were dissolved in 1.0 mL methanol.Then,the solutions were purified by dispersive solid phase extraction with C18,N-(n-propyl)ethylenediamine(PSA)and amino bonded silica gel(NH2)sorbents.The residues were separated on a ZORBAX C18 chromatographic column(50 mm×2.1 mm,1.8μm).The MS data were acquired in positive and negative multiple reaction monitoring(MRM)modes,and quantitated by the isotope internal standard method.With this approach,the linear relationships of the 28 veterinary drugs were good.The linear correlation coefficients were greater than 0.99.The limits of detection and limits of quantification were in the range of 0.1-0.8μg/kg and 0.3-2.7μg/kg,respectively.The average recoveries at three spiked levels(1,5,and 20 fold of the LOQs)were 70.0%-120%with relative standard deviations of 0.9%-10.0%.The method is simple,rapid,accurate and sensitive,and it is suitable for the rapid determination of various prohibited and restricted drugs in fish and shrimp.