UPLC-MS/MS法测定13种中成药中的马兜铃酸A

Determination of aristolochic acid A in 13 kinds of traditional Chinese patent medicines by UPLC-MS/MS

目的采用超高效液相色谱-电喷雾三重四极杆质谱仪测定中药中马兜铃酸A的含量。方法样品用甲醇超声提取后,提取液通过Phenomenex Strata-X-AW小柱(33μm,60 mg,3 mL)净化去除杂质后,用高效液相色谱-三重四级杆串联质谱进行分析;采用Welch XB C18色谱柱(50 mm×2.1 mm,1.8μm),以含0.1%甲酸的10 mmol·L^-1乙酸铵溶液-乙腈为流动相梯度洗脱,采用电喷雾离子源,正离子模式下,多反应监测模式监测,采用标准曲线法计算含量。结果0.991~4.957×10^3μg·L^-1马兜铃酸A与峰面积呈良好的线性关系(r^2=0.9999),定量限为4.9×10^-3 ng,平均回收率为105.1%,RSD=4.86%。结论所用方法灵敏、快速、准确、专属性强,可用于测定中药中的马兜铃酸A。

OBJECTIVE To determine aristolochic acid A in 13 kinds of traditional Chinese patent medicines by UPLC-MS/MS.METHODS After extracted by methanol,the sample extraction solution were purified on Phenomenex Strata-X-AW(33μm,60 mg,3 mL)column,finally analyzed by UPLC-triple quadrupole MS on a Welch XB-C18 column(50 mm×2.1 mm,1.8μm)using mobile phase of 10 mmol·L^-1 acetic acid solution which contains 0.1% formic acid-acetonitrile by gradient elution,and the target compounds were analyzed in the multiple reaction monitoring mode(MRM)with positive electrospray ionization(ESI+).The concentration was calculated by standard curve method.RESULTS The linearity range of aristolochic acid A was 0.991 to 4.957×10^3μg·L^-1(r^2=0.9999).The lower limit of quantity was 4.9×10^-3 ng.The average recovery was 105.1% with the RSD of 4.86%.CONCLUSION The method is sensitive,rapid,accurate and specific for the simultaneous detection of aristolochic acid A in traditional Chinese patent medicines.