HPLC-MS/MS同时检测大鼠体内三聚氰胺和三聚氰酸含量

目的建立HPLC-MS/MS同时检测大鼠血浆和组织中三聚氰胺和三聚氰酸含量的方法。方法以同位素标记的三聚氰胺和三聚氰酸作为内标物,大鼠血浆和组织经二氯甲烷-乙腈前处理后,经色谱柱分离,串联离子阱质谱采用ESI源按运行时间段切换正、负离子模式进行测定。结果在0.3～75μg/ml范围内,三聚氰酸和三聚氰胺具有良好的线性(r≥0.998)。在0.9、5.0、50μg/ml的给药大鼠的血浆样本中,三聚氰酸和三聚氰胺的回收率分别为97.4%～99.4%、96.8%～98.9%,两者的变异系数为2.43%～6.35%(日内)、1.70%～4.12%(日间)和0.30%～9.17%(日内)、0.75%～3.39%(日间);在0.9、5.0、50μg/ml的给药大鼠肝组织样本中,三聚氰酸和三聚氰胺的回收率分别为90.8%～104.6%、91.7%～106.0%,对应的变异系数为3.70%～5.14%(日内)、2.84%～4.50%(日间)和1.31%～2.21%(日内)、1.12%～3.36%(日间)。结论该方法样品前处理简单,基质干扰小,分析效率高,具有良好的精密度、准确度和特异性,是进行三聚氰酸、三聚氰胺在动物体内的组织代谢和毒性研究的理想分析方法。

Simultaneous determination of melamine and cyanuric acid in rat by HPLC-MS/MS

Objective To establish a method for simultaneous determination of melamine and cyanuric acid in rat by HPLC-MS/MS.Methods Using isotope-labelled melamine and cyanuric acid as internal standard,rat plasma and tissue samples were pretreated with dichloromethane-acetonitrile,and separated by chromatographic column switching technique.Detection was carried out by HPLC coupled with electrospray ionization(ESI) working on segment ESI-and ESI+ switching mode.Results The calibration curve of melamine and cyanuric acid were linear(r≤0.998) in the range of 0.3-75 μg/ml.The average recovery rates of melamine and cyanuric acid(added with 0.9,5.0 and 50 μg/ml) in rat plasma were 97.4%-99.4% and 96.8%-98.9%,respectively;while the relative standard deviations(RSD) of the method were 2.43%-6.35%(inter-day),1.70%-4.12%(intra-day),0.30%-9.17%(inter-day) and 0.75%-3.39%(intra-day),respectively.The average recovery rates of rat liver tissue(added with 0.9,5.0 and 50 μg/ml) were 90.8%-104.6% and 91.7%-106.0%,respectively;while the relative standard deviations(RSD) of the method were 3.70%-5.14%(inter-day),2.84%-4.50%(intra-day),1.31%-2.21%(inter-day) and 1.12%-3.36%(intra-day),respectively.Conclusion The method was simple in sample pretreatment,less matrix interference,efficient,precise,accurate and specific.It was ideal to study the mechanism of metabolic behavior and toxicology of melamine and cyanuric acid in animals.