高效液相色谱法同时测定人血浆中氟西汀和喹硫平浓度

Simultaneous determination of the concentrations of fluoxetine and quetiapine in human plasma by HPLC

目的建立同时测定人血浆中氟西汀和喹硫平浓度的高效液相色谱(HPLC)方法。方法采用HPLC同时测定血浆氟西汀和喹硫平浓度。色谱柱为Inertsil ODS-C18柱(4.6 mm×150 mm,5μm),流动相为0.05 mol/L醋酸铵-甲醇(体积比22∶78),流速为0.8 mL/min,检测波长为254 nm,柱温为40℃,萃取剂为乙酸乙酯-二氯甲烷(体积比77∶23),内标物为卡马西平。结果卡马西平和喹硫平、氟西汀色谱峰完全分离,峰形良好,保留时间分别为4.91、6.53和7.48 min,血浆中杂质对测定不产生干扰。氟西汀在10～480μg/L、喹硫平在20～1 600μg/L范围内线性关系良好(r值分别为0.990 5、0.992 9);当信噪比(S/N)=3时,检测限分别为5、8μg/L。氟西汀和喹硫平低、中、高3种浓度绝对回收率均>75%,相对回收率>95%,日内、日间相对标准偏差(RSD)均<10%。结论该方法灵敏、准确、成本低廉,可用于临床氟西汀与喹硫平的血药浓度监测和药物动力学研究。

Objective To establish a method for determining the concentrations of fluoxetine and quetiapine simultaneously in human plasma by high performance liquid chromatography（HPLC）.Methods The plasma concentrations of fluoxetine and quetiapine were determined by HPLC.The column was Inertsil ODS-C18 column（4.6 mm×150 mm,5 μm）,the mobile phase was 0.05 mol/L ammonium acetate-methanol（volume ratio： 22 ∶ 78）,the flow rate was 0.8 mL/min,the detection wavelength was 254 nm,the column temperature was kept at 40℃,the extractant was ethylacetate-dichloromethane（volume ratio： 77 ∶ 23）,and the internal standard was carbamazepine.Results Chromatographic peaks of carbamazepine,quetiapine and fluoxetine were completely separated,and all peak shapes were good.The retention times were 4.91,6.53 and 7.48 min,respectively,and the interference of plasma impurities on the determination did not occur.The calibration curves showed good linearity in range of 10-480 μg/L for fluoxetine and 20-1 600μg/L for quetiapine（r=0.990 5 and 0.992 9）.When the signal-to-noise ratio（S/N）was 3,the detection limits were 5 and 8 μg/L,respectively.The absolute recoveries of fluoxetine and quetiapine in low,middle and high concentrations were 75%,and the relative recoveries were 95%.The relative standard deviation（RSD） of intra-day and inter-day were 10%.Conclusions The method is sensitive,accurate and low-cost,and it can be used for clinical drug monitoring and pharmacokinetics studies of fluoxetine and quetiapine.