HPLC-DAD-MS method for Simultaneous quantitation of flavonoids in Hypericum formosanum and antiglycation activity

Abstract

Hypericum formosanum (HF) is widely used as traditional and folklore medicine in Taiwan. In the present study, ultrahigh performance liquid chromatography equipped with photodiode array detection-mass (UPLC–DAD-MS) method was developed for the separation, identification, and quantification of bioactive flavonoids. The developed method was also validated for accuracy, precision, limit of detection and quantification (LOD and LOQ). In this method, four flavonoids, viz., hyperoside, astiblin, quercitrin, and quercetin, were quantified in linearity range of 10–200 (μg/mL) with correlation coefficient of greater than 0.997. A high recovery (87.55–92.11%) and good reproducibility was obtained for four flavonoids with the relative standard deviation (RSD) ranging from 1.75-2.26 % (intra-day) and 1.99-2.27% (inter-day). Hence, the proposed method for simultaneous quantification of four bioactive flavonoids in the extract and fractions of H. formosanum using UPLC–DAD-MS detection under the specified conditions is accurate and validated. Besides that, ethyl acetate (EtOAc) fraction showed the best anti-glycation and its IC50 was observed at 69.5μg/mL.