HPLC测定氟西汀血药浓度及其临床应用

Determination of fluoxetine hydrochloride in plasma by HPLC

目的建立并优化一种简单、快速、灵敏及高选择性测定氟西汀血药浓度的高效液相色谱法(HPLC),用于临床个体化给药监测。方法血浆样品用乙腈沉淀蛋白法处理。样品分离用Hypersil ODS2(5μm,4.6 mm×200 mm)色谱柱,流动相为乙腈-25 mmol/L磷酸盐缓冲液(pH6.5)(34∶66),流速为1.2 ml/min;进样量为20μl,紫外检测波长227 nm。结果氟西汀的血浆浓度范围是0.5～50 ng/ml,r=0.996 9,低、中、高血浆质控样品日内RSD均小于8.06%,日间RSD均小于10.46%,方法平均回收率分别为91.7%,97.06%和100.75%。结论与已报道的氟西汀血药浓度测定方法相比,本方法血浆样品处理简单,重现性高,选择性好,更经济、污染少且能更好地保护色谱柱。

Objective To establish and optimize a simple, rapid, reliable, high sensitive and high selective method for the determination of fluoxetine hydrochloride in plasma by high-performance liquid chromatographic （HPLC） so as to monitor the clinical drug use. Methods The plasma samples were deproteined by acetonitrile. Separation was performed on a Hypersil ODS2 column（5 um, 4.6 mm× 200 mm）. The mobile phase contained acetonitrile-25 mmol/ L phosphate buffer（ pH6.5 ） （34 ： 66）. The flow rate and sample volume injected were 1.2 ml/min and 20 ul, respectively. Detection wavelength was 227 nm. Results The calibration curve of fluoxetine hydrochloride in plasma was linear in the concentration range of 0.5 - 50 ng/ml and coefficient was 0. 996 9. The intra-assay precision did not exceed 8.06% and inter-assay precision did not exceed 10.46% for low, medium and high quality samples, respectively. The average recovery of the described method was 91.7% ,97.06% and 100.75% , respectively. Conclusion In comparison with previous work, the improved chromatographic method was more economical, less polluted and better protected column, besides simple preparation of plasma samples, high reproducibility, and high selectivity.