超高效液相色谱-Orbitrap高分辨质谱联用快速筛查测定鱼虾贝中79种药物残留

Rapid Screening and Determination of 79 Veterinary Drug Residues in Fish,Shrimp,and Shellfish by Ultra-High-Performance Liquid Chromatography-High Resolution Orbitrap Mass Spectrometry

基于超高效液相色谱-Orbitrap高分辨质谱技术,研发一种同时提取水产样品中多种类兽药残留的溶剂,建立鱼虾贝中11类79种兽药残留的同时快速检测方法。样品中的目标化合物采用混合溶剂[乙腈∶甲醇∶水(3∶1∶1,体积比),内含1%醋酸和10 mmol/L乙二胺四乙酸二钠盐]提取,经Waters HLB固相萃取柱净化;检测时用Waters BEH C18色谱柱分离,2 mmol/L甲酸铵(含0.01%甲酸水溶液)和甲醇梯度洗脱;质谱采用正、负离子同时全扫描/数据依赖的二级扫描模式,在20 min内完成目标化合物的分离和数据采集。建立满足试验要求的79种目标化合物的高分辨质谱库,79种目标化合物在0.5~200.0μg/kg线性相关性良好,相关系数R2均大于0.99。鱼虾贝样品中除头孢唑啉、头孢噻呋、头孢氨苄、哌拉西林、孔雀石绿5种化合物回收率较低外,其余回收率为50%~120%,相对标准偏差为0.69%~19.91%;79种药物定量限为0.2~5.0μg/kg。该法可用于水产品中79种兽药的同时快速筛查和残留测定。

Based on ultra-high-performance liquid chromatography-Orbitrap high-resolution mass spectrometry(UPLC-Orbitrap HRMS)technology,this study developed a solvent for the simultaneous extraction of multiple types of veterinary drug residues in aquatic samples,and established a rapid detection method for 79 types of veterinary drug residues in fish,shrimp,and shellfish.The target compounds in the samples were extracted using a mixed solvent[acetonitrile∶methanol∶water(3∶1∶1,volume ratio)]containing 1%acetic acid and 10 mmol/L ethylenediaminetetraacetic acid disodium salt(EDTA-2Na),followed by purification using a Waters HLB solidphase extraction column.During detection,a Waters BEH C18 chromatographic column was used for separation,with gradient elution using 2 mmol/L ammonium formate aqueous solution(containing 0.01%formic acid)and methanol.MS was performed in positive and negative ion simultaneous full scan/data-dependent secondary scan mode,and the separation and data acquisition of target compounds were completed within 20 min.An HRMS library meeting the experimental requirements was established,and the linear correlation of the 79 target compounds ranged from 0.5 to 200.0μg/kg with good correlation coefficients(R2>0.99).Except for five compounds(cefazolin,ceftiofur,cefadroxil,piperacillin,and malachite green)with relatively low recovery rates in fish,shrimp,and shellfish samples,the recovery rates for the rest ranged from 50%to 120%,with relative standard deviations of 0.69%to 19.91%.The quantification limits for the 79 drugs ranged from 0.2 to 5.0μg/kg.This method could be used for the simultaneous rapid screening and determination of 79 veterinary drugs in aquatic products.