RP-HPLC法测定人血浆和唾液中奥卡西平活性代谢物的浓度

Determination concentration of active metabolite of oxcarbazepine in human plasma and saliva by RP - HPLC

目的建立反相高效液相色谱法测定人血浆和唾液中奥卡西平活性代谢物利卡西平(MHD)浓度的方法学。方法色谱柱为ZORBAX Eclipse Plus C18,柱温为40℃,流速为1 mL.min-1,流动相为甲醇-水=45∶55,检测波长为254nm,内标为对氯苯乙酰胺。结果血浆及唾液中内源性杂质对样品测定无干扰,血浆及唾液中MHD在2～100μg.mL-1内线性关系良好(血浆中γ=0.999 8,唾液中γ=0.999 8),最低检测浓度为2μg.mL-1。血浆中MHD日内、日间RSD均<5%,提取回收率为72%～89%。唾液中MHD日内、日间RSD均<5.6%,提取回收率为75%～91%。结论本方法简便、灵敏、准确,适用于人体内奥卡西平活性代谢物MHD的浓度监测。

Objective To establish a HPLC method for determining the concentration of monohydroxycarbamazepine（MHD） in human plasma and saliva.Methods The analysis was conducted using a ZORBAX Eclipse Plus C18 column and the column temperature was 40 ℃.The mobile phase consisted of methanol and water（45∶55）.The flow rate was 1 mL·min-1 and the detection was performed at 254 nm.And the internal standard substance was p-Chloroacetanilide.Results Endogenous impurities does not interfere with the determination of the samples in plasma and saliva.The standard curve is linear in the range of 2~100 μg·mL-1（γ=0.999 8 in the plasma,γ=0.999 8 in the saliva）,LLOQ is 2 μg·mL-1.The RSDs of inter-day and intra-day are all less than 5% in the plasma and the extraction recovery is 72%~89%.The RSDs of inter-day and intra-day are all less than 5.6% in the saliva and the extraction recovery is 75%~91%.Conclusion This method is simple,sensitive,accurate and suitable for routine determination of the concentration of MHD in human plasma and saliva.