HPLC手性固定相法测定血浆中氟西汀对映体的浓度

目的:建立以高效液相色谱手性固定相法测定血浆中氟西汀对映体浓度的方法。方法:采用Chirobiotic V手性色谱柱,流动相为甲醇-冰醋酸-三乙胺(100∶0.04∶0.04),检测波长为226nm,流速为1mL·min-1,内标为氯氮平。结果:R-去甲氟西汀和S-去甲氟西汀对氟西汀对映体和内标的分离没有干扰。R-氟西汀、S-氟西汀检测浓度分别在8.1~432(r=0.9998)、10.8~432(r=0.9992)ng·mL-1范围内线性关系良好;其相对回收率分别为97.2%~101.9%和98.7%~102.2%,日内RSD分别为2.6%~5.3%和2.1%~6.7%,日间RSD分别为7.7%~8.9%和8.0%~10.6%。结论:本试验所建立的方法简便、可靠、灵敏度高,可用于血浆中R-氟西汀和S-氟西汀的血药浓度测定及对映体的药动学研究。

Determination of Enantiomers of Fluoxetine in Plasma by HPLC-Chiral Stationary Phase

OBJECTIVE： To establish an HPLC-CSP （chiral stationary phase） for the determination of plasma levels of enantiomers of fluoxetine. METHODS： Enantiomers of fluoxetine were separated on a Chirobiotic V column. The mobile phase consisted of methanol - acetic acid - triethylamine （100 ： 0.04 ： 0.04） with a flow rate of 1 mL · min^ -1. The detection wavelength was set at 226 nm.Clozapine was used as the internal standard. RESULTS： Under this chromatographic condition, both R - norfluoxetine and S - norfluoxetine had no interference on the enantiomers of fluoxetine and the internal standard. The calibration curve was linear in the range of 8.1-432 ng · mL^-1（ r = 0.999 8） for R- enantiomer and in the range of 10.8-432 ng · mL^ -1 （ r = 0.999 2） for S - enantiomer. The relative recovery of R - fluoxetine and S - fluoxetine in plasma were 97.2% - 101.9% and 98.7%- 102.2% , respectively. RSD of intraday and interday ranged from 2.6% to 5.3% and from 7.7% to 8.9% for R- fluoxetine, and from 2.1% to 6.7% and from 8.0% to 10.6% for S- fuoxetine, respectively. CONCLUSION： The method is simple, reliable and sensitive, and applicable for the determination and pharmacokinetic study of the enantiomers of fluoxetine in plasma.