超高压液相色谱-静电场轨道阱高分辨质谱筛查渔用饲料中175种药物

Screening 175 veterinary drugs in fishery feed by ultrahigh performance liquid chromatography-orbitrap high resolution mass spectrometry

建立了超高压液相色谱-静电场轨道阱高分辨质谱快速筛查渔用饲料中多种类药物残留的方法。饲料样品经1%甲酸乙腈溶液提取,浓缩复溶后加水稀释,离心除沉淀,以0.1%甲酸水溶液和0.1%甲酸乙腈溶液为流动相,经Accucore RP-MS色谱柱(100 mm×2.1 mm i.d.,2.6μm)梯度洗脱分离,H-ESI(Heated Electrospray Ionization)离子化,采用Full M S/dd-M S2(Top N)模式采集数据,利用自建的数据库进行定性分析。当满足母离子质荷比偏差<3×10-6;保留时间偏差<15 s;同位素质荷比偏差<1×10-5、相对丰度偏差<25%、综合比对>75%;碎片离子质荷比偏差<2×10-5且至少有一个碎片与数据库中匹配时认为确证检出。方法在200μg/kg可同时定性确证175种药物,50μg/kg同时定性确证136种药物,10μg/kg同时定性确证116种药物,RSD<30%,母离子相对偏差均小于1.9×10-6。200μg/kg加标时,81%的化合物回收率在30%~110%之间。

A rapid screening method of 175 veterinary drug residues in fishery feed based on simple and fast pretreatment and ultrahigh performance liquid chromatography coupled to quadrupole-orbitrap-high resolution mass spectrometer （UPLC-Q-Orbitrap HRMS ） was established. In this method, analytes in samples were extracted with 1% （V/V） formic acid in acetonitrile, The extract was then with N2 and redissolved with 1% （V/V） formic acid in aeetonitrile. Clean-up procedure was achieved by adding isometric water and centrifuged at 5℃, 10000 r/min for 10 minutes to discard precipitation. Then, 175 target compounds were separated on an Aecueore RP-MS （ 100 mm × 2.1 mm i.d., 2.6 μm） column, with 0. 1% formic acid in water and 0. 1% formic acid in acetonitrile as mobile phase. An H-ESI ion source was employed and the data were acquired under Full MS/rid-MS2 （TopN）mode with an inclusion list covering 175 compounds. In the process of qualification, the allowed precursor ion m/z tolerance was set at 5 x 10^-6 , RT override window set as ± 15 seconds, allowed product ion m/z tolerance set as 2 ~ 10^-5 because of the existence of some product ions with m/z less than 100 and at least 1 product ion was required for qualification, allowed isotope m/z deviation set as 1 x 10^-5, intensity deviation as 25% and fit threshold as 75%. In our work, 175 compounds belonging to 18 categories were eluted within 20 minutes, and were identified with this method at 200 μg/kg. Also, 136 and 116 compounds were identified at 50 μg/kg and 10μg/kg, respectively.