A simple, rapid and sensitive multi-residue analytical method was developed and validated for 80 veterinary drugs in Oplegnathus punctatus using ultrahigh performance liquid chromatography-Orbitrap high resolution mass spectrometry (LC-HRMS). The analytes belong to 12 different families include benzimidazoles, β-lactams, lincosamides, macrolides, nitromidazoles, quinolones, sulfonamides and trimethoprim, tetracyclines, triphenylmethane dyes, amphenicols, nonsteroidal estrogens and steroid hormones. The sample preparation was optimized base on QuEChERS (quick, easy, cheap, effective, rugged and safe) procedure. A very simple and sufficient preparation procedure without salting-out and complex clean-up process was studied. It had been proved that water in the extract was helpful for extracting hydrophilic compounds and precipitating the lipids during the subsequent cleaning process. In addition, an appropriate percent of methanol was necessary to some analytes. Finally, a mixture of acetonitrile, methanol and water (3:1:1, v/v/v) which include 1% acetic acid and 10 mM ethylenediaminetetraacetic acid disodium salt 2-hydrate was selected as the extraction solvent, and the clean-up step consisted of a low temperature procedure and two times of high-speed centrifugation to deproteinize and remove lipids. The detection and quantification of all compounds were performed by ultrahigh performance liquid chromatography coupled with electrospray ionization quadrupole Orbitrap high resolution mass spectrometry in positive and negative ion mode. This methodology was validated according to the Commission Decision 2002/657/EC and SANTE/11945/2015. The recoveries ranged from 60.74%–109.85% with relative standard deviations (RSDs) < 20%. The limits of quantification (LOQs) were 0.25–25 ug/kg, for the analytes which the MRL or MRPL had been established in fish tissue, the LOQs were all lower than their own legal tolerances. The values of decision limit (CCα) and detection capability (CCβ) were in the range of 1.91–1001.13 ug/kg and 3.52–1002.26 ug/kg, respectively. This validated method has been successfully applied on the determination of veterinary drugs in real commercial oplegnathus punctatus samples.

建立了一种简单，快速，灵敏的多残留分析方法，并验证了马鞭草中80种兽药的有效性。使用超高效液相色谱-Orbitrap高分辨率质谱（LC-HRMS）。分析物属于12个不同的家族，包括苯并咪唑，β-内酰胺，林可酰胺，大环内酯，硝基咪唑，喹诺酮，磺酰胺和甲氧苄氨嘧啶，四环素，三苯甲烷染料，苯甲酚，非甾体雌激素和甾体激素。样品制备基于QuEChERS（快速，简便，便宜，有效，坚固和安全）程序进行了优化。研究了一种非常简单且充分的制备过程，无需盐析和复杂的清理过程。已经证明提取物中的水有助于在随后的清洁过程中提取亲水性化合物和使脂质沉淀。此外，某些分析物需要适当百分比的甲醇。最后，选择乙腈，甲醇和水（3：1：1，v / v / v）的混合物作为萃取溶剂，其中乙腈，甲醇和水的混合物包含1％的乙酸和10 mM的乙二胺四乙酸二钠盐2-水合物，包括低温程序和两次高速离心以去除蛋白质和去除脂质。所有化合物的检测和定量均通过超高效液相色谱结合电喷雾电离四极杆Orbitrap高分辨率质谱在正离子和负离子模式下进行。该方法已根据委员会决定2002/657 / EC和SANTE / 11945/2015进行了验证。回收率范围为60.74％–109.85％，相对标准偏差（RSD）<20％。定量限（LOQ）为0.25–25 ug / kg，对于鱼组织中已建立MRL或MRPL的分析物，LOQ均低于其法律允许的范围。决策极限（CCα）和检测能力（CCβ）的值分别在1.91–1001.13 ug / kg和3.52–1002.26 ug / kg的范围内。该验证方法已成功应用于实际商业中兽药的测定马鞭草样品。