互为内标法测定患者血浆中氟西汀和西酞普兰的浓度

Determination of Fluoxetine and Citalopram in Patient Plasma Using Each Internal Standard Method by HPLC

目的:建立测定人血浆中氟西汀、西酞普兰的高效液相色谱-荧光(HPLC-FLU)法,为临床用药提供参考。方法:氟西汀和西酞普兰互为内标,血浆样品碱化后经液-液萃取,以Ultimate XB-C18反相色谱柱进行分离,流动相为乙腈:0.02 mol·L-1磷酸氢二钠溶液(35∶65,pH 5.0),流速为1.0 ml·min-1,柱温为30℃。荧光检测氟西汀和西酞普兰的激发波长为230nm,发射波长为305 nm。结果:氟西汀、西酞普兰血药浓度均在15.625～1 000μg.L-1范围内线性关系良好;平均回收率分别为100.72%和100.94%;分析方法的最低定量限均为5μg.L-1;日内、日间RSD均<3%;冻融稳定性RSD均<10%。结论:本法快速、简便、灵敏、准确,适用于氟西汀、西酞普兰的血药浓度监测及药动学研究。

Objective： To establish an HPLC method with fluorescence detection assay for the determination of fluoxetine and citalopram in the patient plasma and provide a scientific basis for the clinical application.Method： Fluoxetine and citalopram were used as the internal standard for each other.The samples were alkalized and extracted by liquid-liquid extraction.Then the samples were separated on an Ultimate XB-C18 column with the mixture of acetonitrile-0.02 mol·L-1 disodium hydrogen phosphate（35∶ 65,pH 5.0） as the mobile phase.The flow rate was 1.0 ml·min-1.The column temperature was set at 30℃.Fluoxetine and citalopram were detected by fluorescence detection（Ex=230 nm,Em=305 nm）.Result： Linearity was obtained from 15.625 to 1 000 μg·L-1 for fluoxetine and citalopram.The LOQ was 5 μg·L-1.The average recovery of fluoxetine and citalopram was 100.72% and 100.94%,respectively.Both the inter-day and intra-day RSD were less than 3%.The Freeze-thaw stability RSD was less than 10%.Conclusion： The method is rapid,convenient,sensitive and reliable in the drug determination,adverse reaction control and pharmacokinetic study of fluxetine and citalopram in human plasma.