高效液相色谱-串联质谱法测定动物源性基质中氟喹诺酮及四环素类抗生素残留

Determination of fluoroquinolones and tetracyclines antibiotics residues in animal-derived matrix by high performance liquid chromatography-tandem mass spectrometry

目的建立高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)检测动物源性基质中的氟喹诺酮和四环素类抗生素残留。方法样品经EDTA-Mcllvaine缓冲溶液提取,超低温冷冻离心脱脂,过亲水亲油平衡(hydrophile lipophile balance,HLB)净化柱,采用Agilent C18柱(100 mm×2.1 mm,2.7μm),以0.1%甲酸-甲醇为流动相,用多离子反应监测(multiple reaction monitoring,MRM)模式进行质谱检测,基质曲线外标法定量。结果该方法对以上13种抗生素类化合物的线性范围为0.02~20μg/kg,相关系数(R)大于0.9923;氟喹诺酮类抗生素的检出限为0.01~0.15μg/kg,定量限为0.03~0.40μg/kg,四环素抗生素的检出限为0.08~0.15μg/kg,定量限为0.27~0.40μg/kg。在0.4、2.0、10μg/kg 3个添加水平的回收率为84.1%~105.0%,相对标准偏差为1.15%~7.81%。结论该方法简单、快速、干扰小,适合动物源性基质体系中兽药残留检测。

Objective To establish a sensitive and convenient method for determination of luoroquinolones and tetracyclines antibiotics residues in animal-derived matrix by high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）.Methods EDTA-Mcllvaine buffer solution was used as the extraction solvent,and spiked samples were purified with hydrophile lipophile balance （HLB） column after low temperature freezing centrifugation.Qualitative analysis was carried out by the multiple reaction monitoring （MRM） mode after the Agilent chromatographic separation with C18 column （100 mm×2.1 mm,2.7 μm）,using methanol and 0.1% formic acid water as mobile phase.The matrix curve external standard method was used for quantitation.Results The linear ranges of 13 kinds of antibiotics were 0.02～20 μg/L,with correlation coefficients above 0.9923.The limits of detection （LODs） were 0.01～0.15 μg/kg for fluoroquinolones,and 0.08～0.15 μg/kg for tetracyclines,respectively.The limits of quantitation （LOQs） were 0.03～0.40 μg/kg for fluoroquinolones,and 0.27～0.40 μg/kg for tetracyclines.The recoveries spiked at 3 levels of 0.4,2.0,10 μg/kg were 84.1%～105.0 %,and relative standard deviations （RSDs） were 1.15 %～7.81%.Conclusion The established method is simple,rapid,and sensitive,which is suitable for the determination of veterinary drugs residues in animal-derived matrix