RP-HPLC法测定马兜铃酸含量方法的优化

Study on the Determination of Aristolochic Acid by RP-HPLC

目的优化高效液相色谱法对马兜铃酸进行含量分析。方法通过比较不同提取溶剂、不同提取方法,确定较为合理的供试品溶液制备方法。比较不同流动相条件对马兜铃酸含量测定的影响,选择含量分析色谱条件。测定不同波长下的最低检出限度,确定痕量分析条件。结果供试品溶液制备中以70%甲醇直接加热回流提取,效率优于其他提取方法。做一般含量分析时可以选择流动相为甲醇-1%醋酸水溶液(70∶30)或乙腈-0.3%(NH4)2CO3(pH=7.5)(25∶75),250nm或319nm作为检测波长。做痕量分析时宜选择流动相为乙腈-0.3%(NH4)2CO3(pH=7.5)(25∶75),并选择检测波长为224nm,最小检出量可达0.02ng。结论本实验所建立的方法可以准确可靠的进行马兜铃酸含量分析并进行痕量检查。

Objective To optimize the RP-HPLC procedure for the determination of aristolochic acid. Method Different extraction solvents and methods have been screened to get the most efficient way for sample preparation. The influence of different mobile systems on quantitation were compared to choose an approprate mobile system for deten-nination. Besides, the minimum limits of different detecting waves were measured to give the method to detect trace amount of aristolochic acid. Result Refluxing with 70% methanol was better than other ways in sample preparation. Both methanol-1% acetic acid （70 ： 30） solution and 0.3% ammonium carbonate solution （pH=7.5）-acetonitrile （75 ： 25） with wavelength of 250 nm and 319 nm can be used for quantitation while 0.3% ammonium carbonate solution （pH=7.5）-acetonitrile （75 ： 25） with wavelength of 224 nm for trace detection. The minimum detectable amount was 0.02 rig. The method established can be applied to determinate aristolochic acid and detect trace amout exactly.