反相高效液相色谱/电喷雾质谱法测定血浆中盐酸舍曲林

Determination of Sertraline in Human Plasma by High Performance Liquid Chromatography Coupled with Electrospray Ionization Mass Spectrometry

采用反相高效液相色谱-电喷雾质谱法（LC—ESI／MS）测定人体血浆中盐酸舍曲林。以盐酸丙咪嗪为内标，按内标法定量。血浆样品经pH10．00碳酸钠溶液碱化，加入甲基叔丁基醚萃取，离心分离，取上清液吹干，用流动相溶解进样。色谱柱：RP—Extend—C18（150mm×4．6mm，5μm），柱温：40℃；流动相：pH3．00的三氟乙酸溶液-甲醇（40：60，V／V），流速0．6mL／min。质谱采用选择离子监控模式，检测离子的核质比（m／z）分别是281（丙咪嗪）和159（舍曲林）。舍曲林和丙咪嗪的保留时间分别是5．4min和3．8min；舍曲林标准曲线线性范围为1～80μg/L；检出限为1．0μgL（S／N〉10）；日间、日内相对标准差均小于6．O％，相对回收率为90％～106％；提取回收率在75％～93％范围内。此法适合人体药代动力学的血浆样品的分析，结果准确、可靠。

A method for the determination of sertraline in human plasma using reversed phase high performance liquid chromatography coupled with electrospary ionization mass spectrometry （ LC-ESI/MS）, with the selected ion monitoring （SIM） mode, was described. In this work, imipramine was set as an internal standard. Analytes were extracted from plasma samples that previously were alkalified with pH 10.00 Na2CO3 solution into a methyl tert-butyl ether phase. Supernatant was dried with gas and the residue was dissolved using mobile phase prior to analyzing. Optimized conditions involved separation on am RP-Extend-C18 column （ 150 mm ×4.6 mm, 5 μm） , keeping at 40℃ using washing solution composed of 60% methanol and 40% distilled water （pH 3.00, adjusted with trifluoroacetic acid） at 0.6 mL/min. The analysis was performed in the selected ion monitoring mode at m/z 159 for sertraline and m/z 281 for Imipramine. The retention time of sertraline and Imipramine was observed at 5.4 min and 3.8 min, respectively. The linear response for sertraline was obtained in the range of 1 - 80 μg/L with a correlation coefficient of 0. 9999. The limit of quantification, using 500 μL of plasma, was 1 μg/L （S/N 〉 10）. Precision of either intra-day or inter-day assay were less than 6.0% and relative recovery ranged from 90% to 106%. The exaction recovery was between 75% and 93%. These findings indicate that the assay method has adequate precision and accuracy to determine the amount of Sertraline in human plasma so that it is quite suitable for the analysis of samples collected during pharmacokinetics investigations in humans.