LC-MS/MS同时测定曲美布汀和活性代谢物在Beagle犬体内的血药浓度及其药代动力学研究

目的: 建立灵敏稳定的HPLC-MS/MS方法,用于测定Beagle犬血浆中的曲美布汀(TM)及N-去甲基曲美布汀(N-TM)含量,并研究曲美布汀片剂在健康Beagle犬体内的药代动力学。 方法: 样品处理采用液液萃取方法,选择西布曲明(IS)为内标。进行HPLC-MS/MS分析,采用ESI源,MRM离子检测模式,TM、N-TM和IS的检测离子对分别为m/z 388.2→343.2,m/z 374.2→195.2,m/z 280.3→125.1。单剂量给药300 mg后,测定犬血浆中TM及N-TM的浓度,并计算药代动力学参数。 结果: 血浆中TM及N-TM检测的线性范围分别为1~500 ng·mL^-1 (r=0.9959),0.2~100 ng·mL^-1(r=0.9951),萃取回收率均高于85%,日内、日间RSD均小于10%。犬血浆中TM及N-TM C_max分别为(192±32.1)ng·mL^-1和(47.1±22.6)ng·mL^-1,T_max均为(0.667±0.258)h和(1.08±0.492)h,T_1/2分别为为(10.8±4.27)h和(7.06±2.53)h。 结论: 该方法可简便、高效地测定犬血浆中TM及N-TM浓度,并应用于药代动力学研究。

Objective: To develop an LC-MS/MS method for the simultaneous determination of trimebutine(TM) and N-monodemethyltrimebutine(N-TM) concentration in dog plasma to study the pharmacokinetics of TM maleate in healthy Beagles. Method: The analytes were extracted from plasma samples by liquid-liquid extraction and sibutramine was selected to be the internal standard.Detection was carried out on a triple quadrupole tandem mass spectrometer in multiple reactions monitoring (MRM) mode using positive electrospray ionization (ESI).The ion combination of m/z 388.2→m/z 343.2,m/z 374.2→m/z 195.2,m/z 280.3→m/z 125.1 was used to qualify TM,N-TM and the internal standard,respectively.After a single oral dose of 300 mg TM,the concentrations of TM and N-TM were detected and the pharmacokinetic parameters were calculated by DAS 2.0. Result: The calibration curve was in good linearity over the range of 1-500 ng·mL^-1 (r=0.9959) for TM and 0.2-100 ng·mL^-1 (r=0.9951) for N-TM.The intra-day and inter-day precisions (RSD) were lower than 10% and the extraction recovery was higher than 85% for both determined compounds.The main pharmacokinetic parameters of C_max,T_max and T_1/2 were (192±32.1)ng·mL^-1,(0.667±0.258)h,(10.8±4.27)h for trimebutine(47.1±22.6)ng·mL^-1,(1.08±0.492)h,7.06±2.53 h for N-TM,respectively. Conclusion: The method is shown to be simple,accurate,sensitive and specific for the simultaneous determination of TM and N-TM in Beagles plasma and can be used for the pharmacokinetic study.