基于响应面优化-UPLC-MS/MS测定水产品中孔雀石绿及其代谢物的方法研究

Determination of Malachite Green and Its Metabolites in Aquatic Products Based on Response Surface Optimization-UPLC-MS/MS

本文建立了水产品中孔雀石绿与其代谢物提取的最佳实验条件,并对其在水产品中残留量的检测分析方法进行了研究。在单因素试验的基础上,以硼氢化钾、酸性氧化铝和缓冲体系pH为影响因素,以孔雀石绿和其代谢物的提取含量为响应值进行响应面分析。运用超高效液相色谱-串联质谱仪进行分析,并在不同基质中进行方法学验证。孔雀石绿及代谢物提取的最佳实验条件为:硼氢化钾3 mL,酸性氧化铝7 g,缓冲体系pH=4.2,其提取含量为15.76 ng·g-1,与理论值基本吻合;方法检出限为0.2μg·kg-1,线性范围为0.5~100 ng·mL-1;当阴性基质中的标准物质加入量为12.5~50.0μg·kg-1时,平均回收率为89.3%~101.4%,RSD为2.5%~6.6%。该法高效、快速、简单,可用作对于大批量的水产品中孔雀石绿和其代谢物的快速定性及定量分析。

The best experimental conditions of malachite green and its metabolites extraction in aquatic products were established,and the detection and analysis methods of its residues in aquatic products were studied.On the basis of single factor test,potassium borohydride,acid alumina and buffer system pH were taken as influencing factors,and the extraction content of malachite green and its metabolites was taken as response value for response surface analysis.Ultra high performance liquid chromatography tandem mass spectrometry was used for analysis,and the method was verified in different substrates.The best experimental conditions for extraction of malachite green and its metabolites are:3ml of potassium borohydride,7 g of acid alumina,pH=4.2 of buffer system;the extraction content is 15.76 ng·g-1,which is basically consistent with the theoretical value;the detection limit of the method is 0.2μg·kg-1,and the linear range is 0.5~100 ng·ml;when the amount of standard substance in the negative matrix is 12.5~50.0μg·kg-1,the average recovery is 89.3%~101.4%,RSD is 2.5%~6.6%.The method is efficient,rapid and simple,and can be used for rapid qualitative and quantitative analysis of malachite green and its metabolites in large quantities of aquatic products.