An integrated approach: green extraction technique optimization, mechanistic study, and bioactivity profiling of four...

In this study, a novel strategy for simultaneous extraction of four key flavonoids (epimedin A, epimedin B, epimedin C, and icariin) from Epimedium sagittatum using deep eutectic solvent-ultrasound assisted extraction (DES-UAE) was developed. Initially, 24 kinds of DESs were produced, a proline-lactic acid system demonstrated optimal extraction efficiency. The ideal parameters were established by single-factor experiments and response surface methodology optimization: water content of 29%, molar ratio of 1 : 1.9, liquid-solid ratio of 32 : 1 mL g^-1, ultrasound power of 280 W and extraction time of 30 min, achieving the four target flavonoids yield of 174.69 ± 7.76 mg g^-1. DES-UAE achieved a higher extraction yield than traditional methods. Moreover, the DES could be reused at least five times. Scanning electron microscopy (SEM) analysis of E. sagittatum powders subjected to distinct processing methodologies provided mechanistic insights into the superior performance of DES-UAE. Furthermore, the DES-UAE mechanism was elucidated through the development of molecular dynamics simulation, coupled with molecular interactions to analyze the interaction between IA molecules and DES. Network pharmacology analysis was conducted to screen potential antioxidant target proteins derived from E. sagittatum. Finally, the antioxidant capacity of the extracts was evaluated using three antioxidant experiments, which demonstrated significant radical scavenging activity, underscoring the antioxidant potential of E. sagittatum extracts. In sum, the research developed an eco-friendly and efficient method to extract flavonoids in E. sagittatum.