Study on the Determination of Aristolochic Acid by RP-HPLC / 中国中医药信息杂志

ABSTRACT

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 determination. 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 ng. Conclusion The method established can be applied to determinate aristolochic acid and detect trace amout exactly.