液相色谱-质谱联用法测定消肿止痛酊中马兜铃酸A的含量

Content Determination of Aristolochic Acid A in Xiaozhong Zhitong Tincture by LC-MS/MS

目的:建立测定消肿止痛酊中马兜铃酸A含量的方法。方法:采用液相色谱-质谱联用(LC-MS/MS)法。色谱柱为ZOR-BAXEclipseplusC18柱,流动相为乙腈(含5%水,0.1%甲酸+5mmol/L甲酸铵)-水(0.1%甲酸+5mmol/L甲酸铵)梯度洗脱,流速为0.3ml/min,柱温为40℃,进样量为10μl;通过电喷雾离子化、采用多反应监测方式进行正离子检测,用于定量分析的检测离子为m/z359.1→296.1。结果:马兜铃酸A进样量在11.21～299.0pg范围内与峰面积积分值呈良好的线性关系(r=0.9995);平均加样回收率为119.20%,RSD=4.0%(n=9)。结论:该方法准确、快速、简便、重复性好、干扰少、特异性好,能满足消肿止痛酊中马兜铃酸A的痕量检测要求。

OBJECTIVE：To establish a method for the content determination of aristolochic acid A in Xiaozhong zhitong tincture.METHODS：LC-MS/MS was adopted.The dermination was performed on ZORBAX Eclipse plus C18 column with mobile phase consisted of acetonitrile（containing 5% water,0.1% formic acid,5 mmol/L ammonium formate）-water（0.1% formic acid and 5 mmol/L ammonium formate,gradient elution） at the flow rate of 0.3 ml/min.The column temperature was set at 40 ℃ and injection volume was 10 μl;the detection was performed in multiple reaction monitoring mode（MRM） with positive electrospray ionization（ESI）,by m/z 359.1→296.1 of aristolochic acid A.RESULTS：The linear range of aristolochic acid A were 11.21-299.0 pg（r=0.999 5） with an average recovery of 119.20%（RSD=4.0%,n=9）.CONCLUSIONS：The method is accurate,rapid,simple and reproducible with less interference and good specificity,which can meet the requirements of trace detection of aristolochic acid A.