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

中文关键词:  庆大霉素  新霉素  超高效液相色谱-串联质谱法  鲟鱼

英文关键词:gentamycin  neomycin  ultra performance liquid chromatography-tandem mass spectrometry  sturgeon

中文摘要:

      目的 建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass 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10 mmol/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, 1.6 μm)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(multiple reaction 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25~300 μg/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10 μg/kg, 定量限为25 μg/kg; 添加量为25、50、100 μg/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50~600 μg/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20 μg/kg, 定量限为50 μg/kg; 在50、100、200 μg/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。

英文摘要:

      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.