超高效液相色谱-串联质谱法测定鲟鱼中庆大霉素和新霉素残留量

Determination of gentamycin and neomycin residues in sturgeon by ultra performance liquid chromatography-tandem mass spectrometry

目的建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandemmass spectrometric,UPLC-MS/MS)检测鲟鱼中庆大霉素和新霉素的分析方法。方法样品经10mmol/L磷酸盐缓冲溶液提取, 0.1%甲酸沉淀蛋白, HLB固相萃取柱净化。采用CORTECS HILIC(100 mm×2.1 mm, 1.6μm)色谱柱分离,以0.1%甲酸溶液(A)和乙腈(B)作为流动相进行梯度洗脱,通过电喷雾正离子扫描(electronic spray ion,ESI+),多反应监测模式(multiplereaction monitoring,MRM)对庆大霉素和新霉素的定量离子和定性离子进行测定。结果本方法在5 min内完成2种目标化合物的分离分析。庆大霉素在25～300μg/L浓度范围内呈现良好的线性关系,相关系数r2>0.995;方法检出限为10μg/kg,定量限为25μg/kg;添加量为25、50、100μg/kg时回收率在76.7%～106.3%之间,相对标准偏差(relative standard deviation, RSD)在0.18%～2.15%之间(n=6)。新霉素在50～600μg/L浓度范围内呈现良好的线性关系,相关系数r2>0.996;方法检出限为20μg/kg,定量限为50μg/kg;在50、100、200μg/kg添加水平的回收率为96.1%～109.0%之间,相对标准偏差(RSD)在0.72%～2.84%之间(n=6)。结论该方法精密度好、灵敏度高,能简便、准确地测定鲟鱼中庆大霉素和新霉素的药物残留。

Objective To establish a method for the determination of gentamycin and neomycin residues in sturgeon by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Methods The samples were extracted with 10 mmol/L phosphate buffer solution,precipitating protein with 0.1% formic acid,and purified by HLB solid phase extraction column.And then gentamycin and neomycin residues in the extracts were separated by CORTECS HILIC chromatographic column using a gradient elution program of 0.1% formic acid aqueous solution(A) and acetonitrile solution(B).The quantitative and qualitative ions of gentamicin and neomycin were determined by electronic spray ion(ESI+) and multiple reaction monitoring(MRM).Results The method completed the separation analysis of 2 target compounds within 5 min.Gentamycin had good linear relationships in the range of 25-300 μg/L,r2>0.995.The limits of detection were 10 μg/kg and the limits of quantitation were 25 μg/kg.The recoveries at 3 spiked levels(25,50,100 μg/kg) were 76.7%-106.3%,with the relative standard deviations of 0.18%-2.15%(n=6).Neomycin had good linear relationships in the range of 50-600 μg/L,r2>0.996.The limits of detection were 20 μg/kg and the limits of quantitation were 50 μg/kg.The recoveries at 3 spiked levels(50、100、200 μg/kg) were 96.1%-109.0%,with the relative standard deviations of 0.72%-2.84%(n=6).Conclusion This method is accurate and sensitive,which is convenient for determination of gentamycin and neomycin residues in sturgeon.