高效液相色谱法同时测定人血浆中氯氮平与氟西汀的浓度

Determination of clozapine and fluoxetine in human plasma by HPLC

目的建立同时测定人血浆中氯氮平、氟西汀浓度的高效液相色谱法。方法以DiamonsilTMC18反相柱（150 mm×4.6 mm,5μm）为色谱柱,流动相为30 mmol·L^-1醋酸铵-甲醇（体积比24∶76）,流速为0.8mL·min-1,柱温为40℃,检测波长为254 nm,以乙酸乙酯-二氯甲烷（体积比80∶20）为提取剂。结果氯氮平在25.01 000.0μg·L^-1、氟西汀在10.0480.0μg·L^-1范围内,峰面积与浓度呈良好线性关系;氯氮平、氟西汀的低、中、高3种浓度相对平均回收率分别为100.4%、98.17%、97.89%和101.3%、98.58%、99.12%,提取回收率分别为74.89%、77.93%、79.84%和71.62%、74.72%、79.63%;日内、日间RSD均低于10%（n=5）。氯氮平与氟西汀的检测限均为5.0μg·L^-1。结论该方法灵敏、准确、简单、快速,可用于临床氯氮平与氟西汀的血药浓度监测和药动学研究。

Objective To establish a method for determining clozapine and fluoxetine in human plasma by RP-HPLC.Methods A reverse phase HPLC system was performed on C18 column（150 mm×4.6 mm,5 μm） with the mobile phase consisted of 30 mmol·L^-1 ammonioum-methanol（24∶76）.The flow rate was 0.8 mL·min-1 and the detection wavelength was at 254 nm.Ethyl acetate and dichloromethane（80∶20）was used as extracting solvent.Results The calibration curves were linear in the range of 25.0-1 000.0 μg·L^-1 for clozapine,10.0-480.0 μg·L^-1 for fluoxetine respectively.The average recovery rates of clozapine and fluoxetine in low,middle and high concentrations were 100.4%,98.17%,97.89% and 101.3%,98.58%,99.12%,respectively;the extraction recovery rates were 74.89%,77.93%,79.84% and 71.62%,74.72%,79.63%,respectively.The intra-day and inter-day variation（RSD） was less than 10%（n=5）.The minimum detectable concentration of clozapine and fluoxetine was 5.0 μg·L^-1.Conclusion The method is sensitive,accurate,simple and quick,which can be used for clinical drug monitoring and pharmacokinetics studies of clozapine and fluoxetine.