RP-HPLC同时测定血浆中8种苯二氮类药物

Simultaneous determination of eight kind of benzodiazepines in human plasma by RP-HPLC

目的采用HPLC法建立同时测定血浆中8种苯二氮类药物(氯硝西泮、艾司唑仑、阿普唑仑、三唑仑、澳沙西泮、替马西泮、氯氮和地西泮)的方法。方法血浆样品经正己烷-二氯甲烷(5:3)提取。选用DiamonsilRP-C18柱(250mm×4.6mm,5μm);甲醇-25mmol.L-1醋酸铵溶液(63:37)作流动相,流速1.0mL·min-1,检测波长230nm。结果氯硝西泮、三唑仑、澳沙西泮、氯氮、地西泮的最低检测浓度为8μg·L-1,艾司唑仑、阿普唑仑、替马西泮的最低检测浓度为5μg·L-1;方法回收率接近100%;提取回收率大于70%,日内、日间精密度小于6%;艾司唑仑、阿普唑仑和替马西泮10～1.5×103μg·L-1的线性关系良好,其余5种药物的线性范围为20～1.5×103μg·L-1(r>0.999)。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8种苯二氮类药物的同时定性、定量检测。

OBJECTIVE To establish an HPLC method for simultaneous determination of frequently prescribed eight kind of benzodiazepines（clonazepam,estazolam,alprazolam,triazolam,oxazepam,temazepam,chlordiazepoxide,diazepam） in human plasma.METHODS The reversed-phase C18 column was used and the mobile phase consisted of methanol-25 mmol·L^-1 NH4Ac（63：37） with l mL·min^-1 of flow rate.The absorbance of eluent was monitored at 230 nm.The drugs was extracted with 5 mL of n-hexane：dichloromethane（5：3） from Alkalinized plasma sample and the upper organic layer was evaporated under N2 flow in 50 ℃ water bath.The residue was resolved with 50 μL of mobile phase and 10 μL solution was injected into HPLC system.RESULTS The detection limit of this method was 8 μg·L-1 for clonazepam,triazolam,oxazepam,chlordiazepoxide and diazepam,5 μg·L-1 for triazolam,alprazolam and temazepam,respectively.The method recovery approached 100%,and absolute recovery was more than 70%.The precision（RSD） was less than 6%,and the linearity was found to be 10-1.5 × 10^3 μg·L^-1 for triazolam,alprazolam and temazepam and 20-1500 μg·L^-1 for the other drugs in plasma.Their correlation coefficients were more than 0.999.CONCLUSIONS This method is simple,rapid,accurate,sensitive and selective,and can be used in clinic determination of the eight benzodiazepines in human plasma.