超高效液相色谱-四极杆/静电场轨道阱高分辨质谱快速筛查鱼和虾样品中200种药物残留

建立了超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法，快速筛查鱼和虾样品中200种药物残留。取均质后的鱼或虾样品，加入适量0.1 mol/L EDTA-Na₂溶液，分别用乙腈和乙酸乙酯超声提取，经Oasis PRiME HLB通过式固相萃取柱净化，以0.1%甲酸水溶液和0.1%甲酸-乙腈溶液为流动相，Thermo AccucoreaQ色谱柱(100 mm×2.1 mm×2.6 μm)分离；采用Full MS/dd-MS²模式采集数据，利用自建的包含化合物保留时间、母离子和子离子质荷比等信息的数据库定性，一级质谱全扫描母离子定量。结果表明，本方法的检出限为1~50 μg/kg，在不同的加标水平下，200种药物的加标回收率为30%~120%，精密度为5%~30%，其中，精密度在5%~15%、15%~20%和20%~30%的比例分别为70%、10%和20%，且精密度20%~30%主要分布于低加标水平样品。利用本方法对48个鱼和虾实际样品进行筛查，共定性确定9种药物，分别是乙氧喹啉、恩诺沙星、环丙沙星、氧氟沙星、诺氟沙星、甲氧苄啶、磺胺二甲基嘧啶、敌百虫和依维菌素。该方法前处理简单、可同时测定多种药物、检测效率高、筛查定性准确度高。

Rapid Screening of 200 Drug Residues in Fish and Shrimp by UPLC-Quadrupole/Exactive Orbitrap Mass Spectrometry

To monitor the safety of foods in terms of the restricted or banned compounds, it is especially preferable to develop generic methods which are capable of detecting as many different classes of concerned compounds as possible, with reliable confirm ability, satisfied sensitivity, good time-efficiency and high throughput. In this work, a method of ultra-high performance liquid chromatography (UPLC) quadrupole/exactive orbitrap mass spectrometry was developed for the rapid screening and semi-quantification of 200 drug residues in fish and shrimp. The sample was firstly added with 0.1 mol/L EDTA-Na₂, extracted with acetonitrile and ethyl acetate, cleaned-up by solid-phase extraction using Oasis PRiME HLB column. Then, the target compounds were separated on Thermo AccucoreaQ column (100 mm×2.1 mm×2.6 μm) with a mobile phase consisting of 0.1% formic acid and 0.1% formic acid-acetonitrile. The mass spectrometry data were acquired under Full MS/dd-MS² mode. Self-built database screening containing retention time, precursor ion and product ions were performed for qualification. Them/z of precursor ion, retention time (t_R) and fragment ions (FI) were acquired through analysis, while the isotope pattern for precursor was automatically calculated by TraceFinder software. And then, a database containing m/z of precursor ion,t_R, m/z of FI and isotope pattern were built in TraceFinder. Screening of all the analytes were performed by TraceFinder with the self-built database. The rules of screening were established as follow: allowed m/z deviation of precursor ion was 3×10^-6, allowedt_R deviation was ±15 s, at least one fragment ion match with allowed m/z deviation at 2×10^-5, and the fit threshold for precursor isotope pattern was 75% with allowed mass deviation at 10^-5, and allowed intensity deviation less than 25%. The full scan MS data was used for quantification, and the limits of detection was 1.50 μg/kg. The average spiked recoveries of 200 target compounds were 30%-120% with the relative standard derivations of 5%-30%, 70% of which were 5%-15%, and the higher relative standard deviations of 20%-30% were from samples with low spiked concentration. The developed method was successfully applied to the simultaneous determination of 200 drugs residues in fish and shrimp. The practical application showed that 9 compounds including ethoxyquin, enrofloxacin, ciprofloxacin, ofloxacin, norfloxacin, trimethoprim, sulfamethazine, dipterex, ivermectin were identified in 48 actual fish and shrimp samples. As a qualitative and semi-quantitative screening method, the features of simple pretreatment, simultaneous multiresidue detection, high efficiency and accuracy are demonstrated.