HPLC/MS法测定全血中大隆和氟鼠酮的研究

Determination of brodifacoum and flocoumafen in whole blood by HPLC/MS

目的：建立全血中同时测定大隆、氟鼠酮的高效液相色谱-质谱联用检测方法。方法：血液样品经乙酸乙酯提取后，采用XDBC18柱（150mm×2．1mm×5pm）；流速：500μl/min；柱温：30℃；流动相：甲醇＋0．2％乙酸（88＋12，v／v）；通过电喷雾电离离子化在选择离子监测（SIM）模式下测定，定量分析离子大隆m／z：523[M—H]^-；氟鼠酮m／z：541[M-H]^＋，外标法定量。结果：在0．005-1．0mg／L范围内两者均呈良好的线性关系，大隆、氟鼠酮的平均回收率分别为91．9％和87．6％，RSD分别为5．0％和5．1％，定量检出限分别为0．062和0．051ng。结论：本方法简便、灵敏、重现性好、特异性强，能满足大隆和氟鼠酮中毒病人的诊断要求。

Objective： To establish a HPLC/MS method for determination of brodifacoum and flocoumafen in whole blood. Methods：After brodifacoum and flocoumafen in whole blood were extracted by ethyl acetate, the samples were separated on ZORBAX Eclipse XDB C18 （ 150 mm ×2. 1 mm ×5 μm） at a constant temperature of 30℃ by using a mobile phase consisting of CH3OH（88% ） and 0. 2% CH3COOH（ 12% ）. The flow rate was 500 μl/min. Detection was performanced on an electrospray ionization （ESI） in the negative mode. Quantified ions were selected and their molecular ions, brodifacoum m/z 523 [ M - H]^-, flo- coumafen m/z 541 [ M - H]^ -. Results：Calibration curves were linear within the range of 0. 005 - 1.0 mg/L and the limit of quan- tification was found to be 0. 062 ng and 0. 051 ng for brodifacoum and flocoumafen, respectively. The extraction recoveries were 91.9% and 87.6% and the RSD were 5. 0% and 5.1% for brodifacoum and flocoumafen, respectively. Conclusion：This method is simple, sensitive and accurate for the determination in poisoned patients.