HPLC-MS/MS测定鱼腥草和复方鱼腥草合剂中马兜铃酸类化合物含量

Content Determination of Aristolochic Acid Analogs in Houttuynia Cordata and Compound Yuxincao Mixture by HPLC-MS/MS

关键词:  鱼腥草  复方鱼腥草合剂  马兜铃酸  HPLC-MS/MS

Key words:  Houttuynia cordata  Compound Yuxincao mixture  aristolochic acid analogs  HPLC-MS/MS

摘要:

目的 建立HPLC-MS/MS同时测定鱼腥草药材和复方鱼腥草合剂中马兜铃酸Ⅰ、马兜铃酸Ⅱ和马兜铃内酰胺Ⅰ含量的方法。方法 采用Phenomenex Kinetex C₁₈色谱柱（100 mm×2.1 mm，2.6 μm）；流动相为乙腈和0.2%甲酸水溶液，梯度洗脱，流速为0.3 mL·min^–1，柱温为30℃；采用电喷雾离子源，以多反应离子监测模式进行质谱检测。结果 马兜铃酸Ⅰ、马兜铃酸Ⅱ和马兜铃内酰胺Ⅰ分别在11.91~1 985 ng·mL^–1（r=0.995 3）、16.87~2 812 ng·mL^–1（r=0.995 7）和3.404~567.4 ng·mL^–1（r=0.996 6）内有良好的线性关系。各化合物平均回收率为85%~111%，RSD为0.2%~6.7%。马兜铃酸Ⅰ检测限为2.395 ng·mL^–1、定量限为7.984 ng·mL^–1；马兜铃酸Ⅱ检测限为3.389 ng·mL^–1，定量限为10.50 ng·mL^–1；马兜铃内酰胺Ⅰ检测限为0.334 2 ng·mL^–1，定量限为1.113 ng·mL^–1。结论 该方法专属性好，灵敏度高，准确可靠耐用，可以用于鱼腥草合剂中马兜铃酸Ⅰ、马兜铃酸Ⅱ和马兜铃内酰胺Ⅰ的含量测定。

Abstract:

OBJECTIVE To establish an HPLC-MS/MS method for simultaneous determination of aristolochic acid I, aristolochic acid II and aristolactam I in Houttuynia cordata and Compound Yuxincao mixture.METHODS Phenomenex Kinetex C₁₈(100 mm×2.1 mm, 2.6 μm) column was used for separation. The mobile phase consisted of acetonitrile and 0.2% formic acid solution with gradient elution. The flow rate was 0.3 mL·min^-1and the column temperature was 30℃. Electrospray positive ion source was used for mass spectrometry detection by multiple reaction monitoring mode. RESULTS The liner range of calibration curve was 11.91-1 985 ng·mL^-1(r=0.995 3), 16.87-2 812 ng·mL^-1(r=0.995 7) and 3.404-567.4 ng·mL^-1(r=0.996 6) for aristolochic acid I, aristolochic acid II and aristolactam I. The average recoveries of each component was in the range of 85%-111%, RSD was 0.2%-6.7%. The limit of detection and quantitation for aristolochic acid I were 2.395 ng·mL^-1 and 7.984 ng·mL^-1, for aristolochic acid II were 3.389 ng·mL^-1 and 10.50 ng·mL^-1, and for aristolactam I were 0.334 2 ng·mL^-1 and 1.113 ng·mL^-1. CONCLUSION The method has good sprcificity, sensitivity and accuracy, and it is suitable for the content determination of aristolochic acid I, aristolochic acicl II and aristolactam I in Compound Yuxincao mixture.