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目的采用HPLC同时测定人血浆中地西泮、氯硝西泮、三唑仑、艾司唑仑的浓度。方法血浆样品经氢氧化钠碱化后用乙酸乙酯萃取,氮气吹干,残留物用甲醇溶解后进样。采用Alltech-C18色谱柱,流动相为乙腈-甲醇-水(30:30:40),流速为1.0ml·min-1,检测波长为225nm。结果三唑仑0.05～10.00mg·L-1与峰面积的线性关系良好(r=0.9998),地西泮、艾司唑仑和氯硝西泮0.1～20.0mg·L-1与峰面积的线性关系良好(r=0.9997)。4种成分的方法回收率分别为96.8%、92.6%、97.7%、94.8%;日内和日间RSD均小于10%(n=5)。结论所建方法操作简便、快速、准确、灵敏度高、重复性好,适用于临床上人血浆中4种苯二氮类药物的同时定性、定量检测。
Objective To determine the concentrations of diazepam, clonazepam, triazolam and estazolam in human plasma simultaneously by HPLC. Methods The plasma samples were alkalified by sodium hydroxide and extracted with ethyl acetate. The mixture was dried under nitrogen and the residue was dissolved in methanol and injected. Alltech-C18 column was used. The mobile phase was acetonitrile-methanol-water (30:30:40), the flow rate was 1.0ml · min-1 and the detection wavelength was 225nm. Results The linear relationship between the concentration of triazolam 0.05 ~ 10.00mg · L-1 and the peak area was good (r = 0.9998), and the concentration of diazepam, estazolam and clonazepam was 0.1 ~ 20.0mg · L-1 Good linearity (r = 0.9997). The recoveries of four components were 96.8%, 92.6%, 97.7% and 94.8%, respectively. The intraday and interday RSDs were less than 10% (n = 5). Conclusion The established method is simple, rapid, accurate, sensitive and reproducible. It is suitable for simultaneous qualitative and quantitative detection of four benzodiazepines in clinical human plasma.