LC-MS/MS法测定大鼠血浆中沃替西汀及其羧基代谢产物

本试验建立了大鼠血浆中沃替西汀及其体内羧基代谢物的LC-MS/MS测定方法,并用该法研究其在SD大鼠体内的药动学特征。以氘代沃替西汀为内标,血浆样品经蛋白沉淀法处理后进样测定,色谱柱为Kromasil■C18柱,以含0.1%甲酸的10 mmol/L甲酸铵水溶液(A)∶甲醇(B)为流动相,梯度洗脱,流速为0.3 ml/min,分析时间5 min。采用电喷雾离子源(ESI)、正离子检测、多反应监测模式,沃替西汀及其羧基代谢产物定量离子对分别为m/z 299.1→150.1和m/z329.2→286.3,内标氘代沃替西汀离子对为m/z 302.3→150.0。沃替西汀线性范围为0.3~100.0 ng/ml(r=0.998 6),羧基代谢物线性范围为0.9~300.0 ng/ml(r=0.999 2)。沃替西汀及其羧基代谢物的批内和批间RSD均小于15%。SD大鼠单次口服6 mg/kg沃替西汀后,沃替西汀及其羧基代谢物的t1/2为(2.66±0.35)和(1.76±0.19)h,cmax为(18.60±5.34)和(309.00±84.10)μg/L,AUC0→t为(123.0±28.9)和(1 165.0±236.0)μg·h·L-1。本法快速简便、灵敏准确,适用于沃替西汀及其羧基代谢物的血药浓度测定及药动学研究。

Determination of Vortioxetine and Its Carboxylic Acid Metabolite in Rat Plasma by LC-MS/MS

A LC-MS/MS method for the determination of vortioxetine and its carboxylic acid metabolite in rat plasma was established and validated,which was applied to study the pharmacokinetics of vortioxetine and its metabolite in Sprague-Dawley rats.Deuvortioxetine was treated as an internal standard,and protein precipitation was used to extract the analytes and internal standard,which were separated on a Kromasil?C18 column.The mobile phase consisted of 10 mmol/L ammonium formate solution containing 0.1%formic acid and methanol,chromatographic separation was achieved by gradient elution at a flow rate of 0.3 ml/min for 5 min.The detection was operated using an electrospray ionization(ESI)in positive mode and the multiple-reaction monitoring mode with the transition of m/z 299.1→150.1 monitored for vortioxetine and m/z 329.2→286.3 monitored for its carboxylic acid metabolite.The mass transition of m/z 302.3→150.0 was used to detect deuvortioxetine.The standard curves of vortioxetine and its carboxylic acid metabolite were linear in the concentration range of 0.3-100.0 ng/ml(r=0.9986)and 0.9-300.0 ng/ml(r=0.9992),respectively.Intra-day and inter-day RSDs of both vortioxetine and the metabolite were less than 15%.After SD rats were orally administrated of vortioxetine at 6 mg/kg,the major pharmacokinetic parameters of vortioxetine and its carboxylic acid metabolite were discovered with t1/2 of(2.66±0.35)and(1.76±0.19)h,cmax of(18.60±5.34)and(309.00±84.10)μg/L,AUC0→t of(123.0±28.9)and(1165.0±236.0)μg·h·L-1.This method was simple,fast,sensitive and accurate,which could be used in the determination of vortioxetine and its carboxylic acid metabolite in rat plasma and pharmacokinetic studies.