UHPLC-MS/MS测定天仙藤中5种马兜铃酸类成分

目的 建立天仙藤中马兜铃酸类成分的液质联用检测分析方法。方法 采用Agilent SB-C₁₈(2.1 mm×50 mm,1.8 μm)色谱柱,以乙腈-体积分数0.1%-甲酸溶液为流动相梯度洗脱,流速0.3 mL·min^-1,以多反应监测模式,建立了同时测定天仙藤中马兜铃内酰胺AⅡ(ALAⅡ)、马兜铃酸Ⅳa(AAⅣa)、马兜铃内酰胺FⅠ(ALFⅠ)、马兜铃内酰胺Ⅰ(ALⅠ)和马兜铃酸Ⅰ(AAⅠ)共5种马兜铃酸类成分的液质联用检测方法。结果 ALAⅡ、AAⅣa、ALFⅠ、ALⅠ和AAⅠ分别在1.12~22.34、258.60~10 344.10、0.98~19.52、11.64~232.85、90.70~2 267.41 ng·mL^-1内线性关系良好;平均加样回收率分别为100.9%(相对标准偏差RSD 4.53%)、105.2%(RSD 4.83%)、102.6%(RSD 8.54%)、104.6%(RSD 3.64%)、114.6%(RSD 3.50%)。一共9批样品,其中有4批均检出上述5种马兜铃酸类成分;4批样品均检出除ALFⅠ外的其余4种马兜铃酸类成分;1批样品未检出AAI,检出其余4种马兜铃酸类成分。结论 上述建立的同时测定2种马兜铃酸和3种马兜铃内酰胺成分的液质联用方法经验证准确、可靠,能够用于马兜铃酸类成分的检测与分析。

Simultaneous Determination of Five Aristolochic Acid Analogues in Aristolochiae Herba by UHPLC-MS/MS

OBJECTIVE To establish the determination method for aristolochic acid analogues in Aristolochiae Herba. METHODS An Agilent SB-C₁₈(2.1 mm×50 mm,1.8 μm) column was used with acetonitrile-0.1% formic acid as mobile phase gradiently eluted at a flow rate of 0.3 ml·min^-1. By using multi-reaction monitoring mode(MRM), a UHPLC-MS/MS method was established for simultaneous determination of five aristolochic acid analogues including aristolactam AⅡ(ALAⅡ), aristolactam FⅠ(ALFⅠ), aristolochic acid Ⅳa(AAⅣa), aristolactam Ⅰ(ALⅠ) and aristolochic acid Ⅰ(AAⅠ). RESULTS The linear relationships for ALAⅡ, ALFⅠ, AAⅣa, ALⅠ and AAⅠ were good in the ranges of 1.12-22.34, 258.60-10 344.10, 0.98-19.52, 11.64-232.85, and 90.70-2 267.41 ng·mL^-1, respectively. And their respective average recovery rates were 100.9%(RSD 4.53%), 105.2%(RSD 4.83%), 102.6%(RSD 8.54%), 104.6%(RSD 3.64%), and 114.6%(RSD 3.50%), respectively. Among nine batches of samples, all of the five aristolochic acid analogues were detected in four batches of samples. For the other four batches of samples, there were no ALFI detected and the other four aristolochic acid analogues were all detected. And AAI was no detected in the last one batch of sample, and the other four aristolochic acid analogues were detected. CONCLUSION The established UHPLC-MS/MS method is accurate and reliable. It can be applied for the analysis of aristolochic acid analogues to guarantee the drug safety.