Major isoflavones that are primarily found in soy (Glycine max), red colver (Trifolium pretense), and Kudzu (Pueraria lobata) are daidzein, glycitein, genistein and their respective glucoside derivatives. Methods for the determination of isoflavones in dietary supplements have been established. These standard methods are based on reversed-phase high performance liquid chromatography (RP-HPLC) and ultraviolet and visible light (UV-Vis) spectroscopy detection. Because of the structural similarity and the large number of isoflavones, the method chromatographic run times are more than 70 minutes long.
Recently, we have transferred an isoflavones standard method onto an ACQUITY Arc UHPLC system. The run time of the new method was reduced to 18 minutes per injection cycle. We also demonstrated the benefits of a small but powerful mass spectrometer, the ACQUITY QDa Mass Detector. With the ACQUITY QDa Detector, the time spent in the method transfer and development was greatly reduced, and the risk of interference was significantly lowered.
In this application note, we further shortened the analysis time to 8.1 min, while meeting performance requirements using an ACQUITY H-Class UPLC System. The method performance characteristics of this UPLC method are presented.