HPLC-MS/MS检测尿液中甲卡西酮及其代谢物卡西酮

Determination of methcathinone and cathinone in urine by HPLC-MS/MS

目的:建立尿液中甲卡西酮及其代谢物卡西酮的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。方法:用含内标双苯戊二氨酯(SKF_(525A))的乙腈提取含有甲卡西酮及卡西酮的尿液,经0.22μm微孔有机滤膜过滤,HPLC-MS/MS检测。采用ZORBAX SB-C_(18)色谱柱(50 mm×4.6 mm,1.8μm),以0.2%甲酸乙腈液和0.2%甲酸水溶液为流动相进行梯度洗脱,流速0.3 mL·min^(-1),柱温23℃,电喷雾离子源(ESI),正离子多反应监测模式(MRM)检测。结果:甲卡西酮质量浓度在0.2~3000 ng·mL^(-1)范围内线性关系良好,线性方程为Y=0.000688X+0.00051(r^(2)=0.9991),检测限为0.1 ng·mL^(-1),定量限为0.2 ng·mL^(-1);卡西酮质量浓度在0.5~3000 ng·mL^(-1)范围内线性关系良好,线性方程为Y=0.000687X+0.000175(r^(2)=0.9988),检测限为0.2 ng·mL^(-1),定量限为0.5 ng·mL^(-1);回收率分别为91.2%~94.1%和91.2%~95.1%,精密度均小于15%,相对偏差均在±15%范围内。结论:该方法操作简便,检测灵敏度较高,可用于尿液中甲卡西酮及其代谢物卡西酮的检测。应用该方法测定实验动物尿液中甲卡西酮及卡西酮含量,结果稳定,方法可靠。

Objective:To develope a high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for determine methcathinone and its metabolite cathinone in urine.Methods:The urine sample containing methcathinone and its metabolite cathinone was extracted with acetonitrile containing internal standard proadifen hydrochloride(SKF_(525 A)).The extract was filtered with 0.22μm organic filter membrane and detected by HPLC-MS/MS.ZORBAX SB-C_(18)column(50 mm×4.6 mm,1.8μm)was used with column temperature of 23℃,mobile phase of 0.2%formic acid aqueous and 0.2%formic acid acetonitrile solution,gradient elution,flow rate of 0.3 mL·min^(-1),electrospray ion source(ESI),positive ion multiple reaction monitoring mode(MRM)detection.Results:The linear relationship of methcathinone was good in the range of 0.2-3000 ng·mL^(-1),and the linear equation was Y=0.000688X+0.00051(r~2=0.9991)with detection limit 0.1 ng·mL^(-1).The calibration curves of cathinone in urine displayed good linearity in the range of 0.5-3000 ng·mL^(-1),and the linear equation was Y=0.000687X+0.000175(r~2=0.9988)with detection limit 0.2 ng·mL^(-1).Their recoveries ranged from 91.2%to 94.1%and 91.2%to 95.1%,respectively.The precision was less than 15%,and the relative deviation was within±15%.Conclusion:This method is simple and sensitive enough to be applied to determine methcathinone and its metabolite cathinone in urine.The experimental results show its reliability in determine methcathinone and cathinone in urine of experimental animal.