虹鳟鱼中孔雀石绿、结晶紫及其代谢物残留量的超高效液相色谱-串联质谱测定法

Determination of malachite green,crystal violet and their metabolite residues in rainbow trout by ultra high performance liquid chromatography-tandem mass spectrometry

目的建立超高效液相色谱-串联质谱法测定虹鳟鱼中孔雀石绿、结晶紫、隐性孔雀石绿和隐性结晶紫残留量。方法对基质效应进行考察,并对超高效液相色谱-串联质谱仪条件进行优化。虹鳟鱼样品用乙腈进行提取,中性氧化铝小柱净化,基质匹配工作曲线内标法定量。结果 4种物质均产生基质抑制效应,基质对孔雀石绿、结晶紫和隐性结晶紫产生的基质抑制效应>-20%,且孔雀石绿和结晶紫的基质抑制效应>-60%,基质效应无法忽略。该方法检出限为0.1~0.2μg/kg,回收率在80.2%~117.0%之间,精密度范围为3.3%~9.6%,相关系数≥0.999 0。结论本方法操作简便、检出限低、灵敏度高、回收率满足方法学要求,适用于虹鳟鱼中孔雀石绿、结晶紫、隐性孔雀石绿和隐性结晶紫残留量的同时测定。

Objective To establish a method for the determination of malachite green(MG),crystal violet(CV),leucomalachite green(LMG) and leucocrystal violet(LCV) residues in rainbow trout by ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Methods Matrix effects were studied and the instrument conditions of the ultra high performance liquid chromatography-triple quadrupole mass spectrometer were optimized. The rainbow trout samples were extracted by acetonitrile,cleaned-up with neutral alumina column,quantified by matrix matched calibration curve with isotope internal standards. Results All four compounds produced matrix suppression effects. The matrix suppression effects of MG,CV and LCV were >-20%,especially MG and CV were more than-60%,which cannot be ignored. The limit of detection(LOD) was 0.1-0.2 μg/kg,the recoveries between 80.2% and 117.0%. The precision range was 3.3%-9.6% and the correlation coefficients were ≥0.999 0.Conclusion This method has the advantages of simple operation,low detection limit and high sensitivity,and recovery are all satisfied with the methodological requirements,which is suitable for the simultaneous determination of MG,CV,LMG and LCV residues in rainbow trout.