液相色谱串联质谱法测定人血浆中文拉法辛及其代谢物的浓度

目的建立测定血浆中文拉法辛及其代谢物O-去甲基文拉法辛的液相色谱串联质谱(LC-MS/MS)方法。方法血浆样品中加入内标(氘6-文拉法辛和氘6-O-去甲基文拉法辛),直接沉淀法处理样品。色谱柱为CAPCELL PAK C18 MGⅢ分析柱(100 mm×2.0 mm,5μm),流动相为含0.3%甲酸的水溶液-含0.3%甲酸的乙腈溶液(78∶22,V/V),流速为0.3 mL.min-1。正离子多离子反应监测(MRM)扫描分析,离子通道分别为m/z 278→58(文拉法辛)、m/z 264→58(O-去甲基文拉法辛)、m/z 284→58(氘6-文拉法辛)、m/z 270→58(氘6-O-去甲基文拉法辛)。结果文拉法辛和O-去甲基文拉法辛的线性范围均为2～1 000μg.L-1,定量下限均为2μg.L-1,提取回收率在90.14%～97.33%,批内、批间RSD均小于8%。结论本方法操作简便,特异性强,灵敏度高,可用于人血浆内文拉法辛和O-去甲基文拉法辛的含量测定研究。

Determination of venlafaxine and its metabolite O-desmethylvenlafaxine in human plasma by LC-MS/MS method

AIM To establish a LC-MS/MS method for the determination of venlafaxine and O-desmethylvenlafaxine in human plasma.METHODS After addition of venlafaxine-D6(internal standard,IS)and O-desmethylvenlafaxine-D6(IS)combination,the analyte was processed by protein precipitation.The chromatographic separation was performed on CAPCELL PAK C18 MGⅢ(100 mm × 2.0 mm,5 μm)column with a mobile phase of 0.3% formic acid water-0.3% formic acid acetonitrile(78 ∶ 22,V/V)at a flow rate of 0.3 mL·min-1.The protonated ions of analytes were detected in positive ionization by multiple reaction monitoring mode(MRM).The mass transition pairs of m/z 278→58,m/z 264→58 and m/z 284→58,m/z 270→58 were used to detect venlafaxine,venlafaxine-D6 and O-desmethylvenlafaxine,O-desmethylvenlafaxine-D6,respectively.RESULTS The linear concentration range for venlafaxine and O-desmethylvenlafaxine were both 2-1 000 μg·L-1.The limit of quantitation for both was 2 μg·L-1.The extraction recovery was 90.14%-97.33%.Within-batch RSD and between-batch RSD were less than 8%.CONCLUSION This method is simple,specific and sensitive,and suitable for determination studies of venlafaxine and O-desmethylvenlafaxine in human plasma.