生物检材中乌头碱的LC-MS/MS快速分析

目的应用高效液相色谱-质谱法对生物检材中乌头生物碱等有毒成分进行快速分析。方法取全血样品经乙腈-甲醇(5:1 v/v)提取,使用Agilent Zorbax SB C18(2.1 mm×50 mm,1.8μm)色谱柱,以0.1%甲酸溶液-乙腈(60:40 v/v)为流动相等度洗脱。在多反应监测模式下测定全血样品中乌头生物碱等有毒成分。结果乌头碱、次乌头碱和中乌头碱的保留时间为0.73 min、0.77 min和0.63 min;用于定量分析的离子对分别为m/z 646.4→586.4(乌头碱)、616.1→556.5(次乌头碱)和632.4→572.1(中乌头碱)。乌头碱在0.1~250 ng/m L内线性关系良好,相关系数(r)≥0.9987,最低检出限0.1ng/m L,精密度考查其变异系数(CV)5.42%(n=6),血液中乌头碱提取回收率不小于90%。结论本文建立的高效液相色谱-质谱法快速、简便、灵敏,适用于天然药毒物检验。

Fast determination of aconitine in biological materials by LC-MS/MS

Objective To establish a HPLC-MS method for determination of aconitum alkaloids in biological samples. Methods The aconitum alkaloids were extracted from the whole blood by using acetonitrile-methanol (5:1 v/v) and then analyzed using HPLC-MS in multiple reaction monitoring (MRM) mode with positive ionization. The analytical column was Agilent Zorbax SB C18 (2.1mm×50mm, 1.8μm)and the mobile phase were water containing 0. 1 % formic acid : acetonitrile (60 : 40 v/v) in isocratic elution. Results The retention time of detection of the aconitine, hypaconitine and mesaconitine were 0.73 min, 0.77 min and 0.63 min, and the precursor product ion combinations of m/z 646.4 → 586.4, 616.1 → 556.5 and 632.4 → 572.1 were used for quantitative analysis, respectively. Calibration curve was linear within the range of 0.1-250 ng/mL with the LOD was 0.1ng/mL, and the coefficient of variation (CV) less than 5.42 % (n=6). The extraction recoveries of aconitine in blood were more than 90 %.Conclusion The results demonstrated that the present method was reliable and robust for natural drugs.