UPLC-MS/MS同时测定京制咳嗽痰喘丸中马兜铃酸Ⅰ、Ⅱ的含量

Simultaneous Determination of Aristolochic Acid Ⅰ and Ⅱ in Jingzhi Kesou Tanchuan Pills by UPLC-MS/MS

目的:建立超高效液相色谱-质谱法(UPLC-MS/MS)同时测定京制咳嗽痰喘丸中马兜铃酸Ⅰ、Ⅱ的含量方法。方法:采用Agilent Poroshell SB-C_(18)色谱柱(100 mm×2.1 mm,2.7μm),以甲醇(A)-5 mmol/L甲酸铵(含0.1%甲酸),(B)为流动相进行梯度洗脱,流速为0.3 mL/min,柱温为30℃,进样量为1μL。质谱采用电喷雾离子源(ESI),正离子模式下多反应监测(MRM),标准曲线法测定。结果:马兜铃酸I和马兜铃酸Ⅱ分别在1.052 4~210.48 pg(R^(2)=0.998 1,n=6)、1.002~100.2 pg(R^(2)=0.999 9,n=6)范围内线性关系良好,平均加样回收率分别为96.09%(RSD=2.08%)和88.05%(RSD=2.64%)。结论:该方法简便、灵敏、专属性好,可用于京制咳嗽痰喘丸马兜铃酸Ⅰ、Ⅱ的含量测定。

Objective:To establish an ultra-high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) method for simultaneous the determination of aristolochic acid Ⅰ and Ⅱ in Jingzhi Kesou Tanchuan Pills.Methods:The UPLC-MS/MS assay was performed on a Agilent Poroshell SB-C_(18) Column(100mm×2.1mm,2.7μm) with methanol(A)-5mmol/L ammonium acetate and 0.1% formic acid in water(B) as mobile phase by gradient elution.The flow rate was set at 0.3mL/min.The column temperature was set at 30℃ and the injection volume was 1μL.Mass spectrometry was carried out on electrospray ionization(ESI) in positive multiple reaction monitoring modes.The standard curve method was used in the experiment.Results:The standard curves of aristolochic acid Ⅰ and Ⅱ showed good linearity in the ranges of 1.052 4~210.48pg,(R^(2)=0.998 1,n=6),1.002~100.2pg(R^(2)=0.999 9,n=6),respectively.Their recoveries were 96.09%(RSD=2.08%) and 88.05%(RSD=2.64%),respectively.Conclusion:The established method was simple,sensitive and specific,which can be used for the limit determination of aristolochic acid Ⅰ and Ⅱ in Jingzhi Kesou Tanchuan Pills.