LC-MS/MS法同时测定曲马多和代谢物O-脱甲基曲马多的血药浓度及其药动学研究

Simultaneous Determination of Tramadol and Metabolite O-demethyl Tramadol in Plasma by LC-MS/MS and Pharmacokinetic Study

目的：建立同时测定术后患者应用曲马多后的曲马多和活性代谢物O-脱甲基曲马多（M1）血药浓度的方法。方法：血浆样品50μ1以乙腈溶液150μl（含内标氟康唑）沉淀蛋白处理后，采用液．质联用（LC-MS／MS）法进样测定，色谱柱为zorbaxSB-C18，流动相为O．1％甲酸．5mmoVL乙酸铵溶液．0．1％甲酸乙腈溶液（梯度洗脱）；用于定量分析的离子反应m／z分别为曲马多264．2→58．6、M1250．0→58．6、氟康唑307→220。结果：2种待分析物血药浓度均在5～1000ng／ml范围内线性关系良好，日内、日间RSD均〈10％；方法回收率分别为92．26％～100．01％、88．45％～100．60％。结论：本方法准确、快速、灵敏，可用于曲马多和M1的血药浓度测定及药动学研究。

OBJECTIVE： To establish the method for simultaneous determination of tramadol and active metabolite O-demethyl tramadol （MI） ill the plasma of postoperative patients treated with tramadol. METHODS： Tramadol and M1 in plasma were deter- mined simultaneously by LC-MS/MS after the plasma sample 50 μl was precipitated with acetonitrile 150 μl containing fluconazol. The determination was performed on Zorbax SB-Cls with mobile phase consisted of 0.1% formic acid-5 mmol/L ammonium acetate solution-0.1% formic acid acetonitrile solution （gradient elution） ; m/z was 264.2→58.6 for tramadol, 250.0→58.6 for Mt and 307→220 for fluconazol. RESULTS： The linear range of tmmadol and M1 both were 5-1 000 ng/ml; RSDs of intra-day and inter-day were lower than 10% ; method recoveries were 92.26%-100.01% and 88.45%-100.60%. CONCLUSIONS： The method is accurate, rapid and sensitive, and can be used for the determination of plasma concentration of tramadol and Mx and pharmacokinetic study.