定量比格犬血浆中去甲文拉法辛LC-MS/MS方法的建立及比格犬血浆中富马酸去甲文拉法辛药代动力学研究

A LC-MS / MS Assay for the Quantification of O-Desmethylvenlafaxine in Beagle Plasma and Applications to Preclinical Pharmacokinetic Studies on Desvenlafaxine Fumarate Administered to Beagles

目的:建立一种快速灵敏的液质联用方法定量比格犬血浆中去甲文拉法辛,并进行比格犬体内富马酸去甲文拉法辛药代动力学研究,同时与琥珀酸去甲文拉法辛比格犬体内药代参数进行对比。方法:样品处理采用叔丁基甲醚进行液液萃取,液相分离采用Agilent Eclipse Plus C18色谱柱(2.1 mm×100 mm,3.5μm),以乙腈-乙酸铵缓冲液(1 mmol/L)(体积比80∶20)进行等度洗脱。采用单次给药和多次给药试验进行比格犬体内药代动力学研究。结果:此方法具有良好的线性、精密度、准确度,定量下限(0.5 ng/mL)高于文献报道。液液萃取的样品提取方法简便,液质联用分析时间相对较短(3 min)。结论:此方法可成功地应用于比格犬血浆中去甲文拉法辛定量分析。比格犬体内富马酸去甲文拉法辛的药代参数(单次给药和多次给药试验)与对照品琥珀酸去甲文拉法辛没有明显的统计学差异。

Objective： To establish a highly sensitive liquid chromatography-tandem mass spectrometry（LC-MS / MS） method for the quantification of O-desmethylvenlafaxine（ODV） in beagle dog plasma and applied to a phar-macokinetic study of desvenlafaxine fumarate（DVF） in beagles,while made a pharmacokinetics comparation with desvenlafaxine succinate（DVS）.Methods： Sample pretreatment includes liquid-liquid extraction by adding tert-butyl methyl ether to the plasma sample containing the internal standard.HPLC separations were performed on an Agi-lent Eclipse Plus C18 column（2.1 mm × 100 mm,3.5 μm i.d.）.Samples were eluted using a mobile phase com-posed of acetonitrile and 1 mmol / L ammonium acetate in water（80∶20 V / V）.Single-time studies and multi-time studies were applicated to pharmacokinetic study.Results： The method has high linearity,precision and accuracy,and a sufficiently low detection limit（0.5 ng / mL） which was lower than the limit of previous reports.The liquid-liquid extraction process was simple and the LC-MS / MS run time is relatively short（3 min）.Conclusion： This method was successfully applied to the determination of ODV concentration in beagle plasma.The pharmacokinetic parameters of DVF in beagle plasma（single-time and multiple-time studies） showed no significant differences from those of DVS.