Development and Validation of an HPLC-DAD Method for Simultaneous Quantitation of Steppogenin and Flavonoids from the Stems of Morus alba

Morus alba L. is well-known for its medicinal and economic value, particularly in Asian countries. Among the isolated compounds from this plant, steppogenin is exhibited as a flavonoid with promising pharmacological properties. This study focused on isolating bioactive compounds, notably steppogenin, from the ethyl acetate extract of M. alba. Additionally, a high-performance liquid chromatography-diode array detector (HPLC-DAD) method for the simultaneous quantification of steppogenin and isolated compounds was developed and validated. The calibration curve showed excellent linearity, with a correlation coefficient (R²) value greater than 0.9957. The limit of detection (LOD) ranged from 0.006 to 0.018 μg/mL, whereas the limit of quantification (LOQ) ranged from 0.020 to 0.061 μg/mL. In precision tests conducted intra-day and inter-day, the accuracy was between 97.32% and 106.39%, with relative standard deviations (RSD) less than 2.27% and 1.65%, respectively. The presence of steppogenin and other flavonoids was confirmed by the study, contributing to the understanding of the chemical composition of M. alba. This validated analytical method offers a reliable means of quantifying steppogenin and aiding future research into its therapeutic potential.