HPLC法同时测定人血浆中6种抗抑郁药的浓度

Simultaneous Determination of 6 Antidepressants in Human Plasma by HPLC

目的:建立一种快速、灵敏的同时测定人血浆中6种抗抑郁药浓度的高效液相色谱法。方法:以异丙嗪为内标,样品碱化后液-液萃取,用Phenomenex-C18反相色谱柱进行分离,以乙腈-0.05mol.L-1磷酸二氢钠溶液(用磷酸调节pH=2.5)为流动相,采用梯度洗脱;流速为1.2mL.min-1,柱温为40℃;荧光检测文拉法辛,激发波长为276nm,发射波长为598nm;紫外检测多塞平、帕罗西汀、舍曲林、氟西汀和阿米替林,检测波长为200nm。结果:文拉法辛、多塞平、帕罗西汀、舍曲林、氟西汀、阿米替林血药浓度分别在5～1000、4～200、20～800、20～800、40～1000、10～400μg.L-1范围内线性关系良好,r均≥0.990;日内、日间RSD均<15%,提取回收率均>60%,方法回收率均>90%。结论:本方法简便、快速、经济、准确、灵敏,可用于血药浓度的临床监测、中毒药物分析及药动学研究。

OBJECTIVE： To establish a rapid and sensitive HPLC method for simultaneous determination of plasma concentrations of 6 antidepressants. METHODS： With diprozin as internal standard, the alkalized samples were extracted by liquid - liquid extraction and separated on a Phenomenex - C18 column using acetonitrile - 0.05 mol·L^-1sodium dihydrogen phosphate（pH was adjusted to 2.5 by phosphoric acid） as mobile phase at a flow rate of 1.2 mol·L^-1 by a gradient elution. The column temperature was set at 40℃. Venlafaxine was detected by fluorescence detection at an Ex wavelength of 276 nm and Em wavelength of 598 nm; doxepin, paroxetine, sertraline, fluoxetine and amitriptyline were detected at an UV detection wavelength of 200 nm. RESULTS： The linear ranges of venlafaxine, doxepin, paroxetine, sertraline, fluoxetine and amitriptyline were 5-1000μg·L^-1, 40200μg· L^-1 20-800μg·L^-1, 40-1000 μg·L^-1 and 10-400μg·L^-1, respectively, with correlation coefficients ≥0. 990 for all. Both the intra - day RSD and inter - day RSD were less than 15% ; the extraction recovery rates were greater than 60% and methodological recovery were greater than 90% for all the samples. CONCLUSION： The method is simple, economical, rapid, accurate and sensitive, and it is applicable for the clinical monitoring of plasma drug concentrations as well as the analysis and pharmacokinetic study of toxic drugs.