A new method using direct on-line coupling between HPLC and UV-DAD/MS was developed for the rapid detection of aristolochic acid I in plant preparations. The use of both UV-DAD and MS detectors enables this method to be very selective. A ng range detection limit was determined for both UV and MS detections. A quantitative determination of aristolochic acid I in plant preparations was achieved using UV and MS detection. A thin layer chromatography assay was also developed for the preliminary detection of aristolochic acid I in complex mixtures.