Determination of hyperoside and 2″-acetylhyperoside from Ligularia fischeri by high performance liquid chromatography

Abstract Ligularia fischeri and its main flavonoids, hyperoside and 2″-acetylhyperoside, posses antioxidant properties. This study was carried out to investigate the contents of hyperoside and 2″-acetylhyperoside in L. fischeri by using high performance liquid chromatography (HPLC). An HPLC–photodiode array (PDA) detection method was established for the simultaneous determination of hyperoside and 2″-acetylhyperoside in L. fischeri . Two flavonoids were successfully separated in less than 20 min using an YMC RP 18 column. The mobile phase was composed of water (A) and acetonitrile (B) with isocratic elution system (23% B) at a flow rate of 1 mL/min. Their calibration curves showed good linear regression ( r  > 0.9992) within the test ranges. The method was validated for specificity, accuracy, precision, and limits of detection. The determined two compounds were well separated with a linear range of 18–180 μg/mL. The contents of hyperoside and 2″-acetylhyperoside were 0.387 ± 0.002 and 0.526 ± 0.006 mg/g in L. fischeri , respectively.