UPLC-MS/MS同时测定通迪胶囊中的马兜铃酸Ⅰ和马兜铃酸Ⅱ

Simultaneous Determination of Aristolochic AcidsⅠandⅡin Tongdi Capsules by UPLC-MS/MS

目的:建立通迪胶囊中马兜铃酸Ⅰ(AA-Ⅰ)和AA-Ⅱ的超高效液相色谱-质谱法。方法:采用Shiseido Shim-pack velox C18色谱柱(100 mm×2.1 mm,1.8μm),以乙腈(A)-0.1%甲酸水溶液(含甲酸铵,B)为流动相梯度洗脱,流速为0.3 mL·min^(–1),采用多反应监测,AA-Ⅰ以m/z 358.9→298.0(定量)和m/z 358.9→296.0(定性)为监测离子对;AA-Ⅱ以m/z 329.0→268.0(定量)和m/z 329.0→294.0(定性)为监测离子对,建立了液相色谱-质谱法检测方法,并对样品进行分析。结果:AA-Ⅰ在1.0524~420.9600 pg、AA-Ⅱ在1.002~100.200 ng时与峰面积线性关系良好(r>0.999),加样回收率分别为97.63%、92.34%,RSD分别为2.24%、1.49%,15批样品中AA-Ⅰ和AA-Ⅱ的质量分数分别为3.760~20.790、0.010~0.156μg·g^(–1)。结论:该方法灵敏、快速、准确、专属性强,可用于测定通迪胶囊中AA-Ⅰ和AA-Ⅱ的含量,可为通迪胶囊用药安全提供参考。

Objective:To establish an ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS/MS)method for the determination of aristolochic acidsⅠand aristolochic acidsⅡin Tongdi Capsules.Methods:A Shiseido Shim-pack velox C18 chromatographic column(100 mm×2.1 mm,1.8μm)was used with a gradient elution of acetonitrile(A)and 0.1%formic acid(with ammonium formate,B)as the mobile phase at a flow rate of 0.3 mL·min^(–1).Multiple reaction monitoring(MRM)was employed with aristolochic acidⅠmonitored at m/z 358.9→298.0(quantitative)and m/z 358.9→296.0(qualitative),and aristolochic acidⅡat m/z 329.0→268.0(quantitative)and m/z 329.0→294.0(qualitative).The UPLC-MS method was established for the detection of aristolochic acidsⅠand aristolochic acidsⅡand applied to sample analysis.Results:Aristolochic acidⅠand aristolochic acidⅡshowed good linearity of injection volume with peak area at 1.0524-420.9600 pg and 1.002-100.200 ng,respectively(r>0.999).The spiked recovery rates were 97.63%and 92.34%with RSDs of 2.24%and 1.49%,respectively.In 15 batches of samples,the mass fractions of aristolochic acidⅠand aristolochic acidⅡranged from 3.760 to 20.790μg·g^(–1) and 0.010 to 0.156μg·g^(–1),respectively.Conclusion:This method is sensitive,rapid,accurate,and highly specific,making it suitable for determining the content of aristolochic acidⅠand aristolochic acidⅡin Tongdi Capsules,which can provide a reference for medication safety.