高效液相荧光色谱法测定人血浆中唑吡坦的浓度

Determination of Zolpidem in Human Plasma by HPLC with Fluorescence

目的:建立测定人血浆中唑吡坦浓度的高效液相荧光色谱法。方法:血浆样品经甲醇沉淀后,以甲醇-1%醋酸液(40:60)为流动相,流速为0.3 ml·min^(-1),色谱柱为Agilent Zorbax C_(18)(150 mm×3 mm,3.5μm),柱温为22℃,荧光检测波长:激发波长(λex)254 nm,发射波长(λem)390 nm。结果:血浆内源性杂质不干扰待测物测定,唑吡坦的线性范围为2～300μg·L^(-1),定量下限为2μg·L^(-1),日内、日间RSD均<5%。样品经3次冻融及再沉淀后,4℃下6 h内稳定性良好。结论:该法灵敏、快速、准确,操作简便,线性范围宽,可用于唑吡坦的临床药动学研究。

Objective： To establish an HPLC with fluorescence method for determination of Zolpidem in human plasma. Method： Plasma samples were treated by methanol. The Agilent Zorbax C18 column （150 mm × 3 mm,3.5μm） at 22℃ was used. The mobile phase consisted of methanol： 1% acetic acid （40： 60） pumped at a flow rate of 0.3 ml· min^-1. The fluorescence detector was opreated at an excitation and emission wavelengths 254 nm and 390 nm, respectively. Result： The intrinsic impurity of plasma did not interfere with the determination of analyte. The linear range of Zolpidem was 2 -300μg· L^-1. Lower limit of quantification was 2μg· L^-1. The precision of inter-day were less than 5%. The stability of unprocessed and processed samples stored at 4 ℃ for 6 hours was good. After 3 cycles of freeze and thaw processes, the plasma samples were stable. Conclusion： The established HPLC-FLU method is fast, sensitive, and accurate and can be applied to the clinical pharmaeokinetics investigations of Zolpidem.