A fast and reliable HPLC method for the simultaneous separation of anthocyanins and flavonols in lotuspetals was developed based on the study of four candidate solvent systems. Fifteen flavonoids were identifiedby high-performance liquid chromatography with photodiode array detection/mass spectrometry.Among them, two anthocyanins and nine flavonols were discovered in lotus petals for the first time. Thiswork is valuable for both the hybrid breeding on lotus oriented to flower color and the utilization of lotuspetals as functional food materials.