Feed

Hedera helix (HE) contains diverse bioactive constituents, including triterpenoid saponins, flavonoids, and phenolic acids, which exhibit various pharmacological activities. In this study, ultrasound-assisted extraction (UAE) combined with natural deep eutectic solvent (NADES) was employed to enhance the extraction efficiency and elucidate the underlying mechanisms. Among the tested formulations, a ternary system composed of malonic acid (Mal), N,N'-dimethylurea (DMU), and 1,4-butanediol (1,4-BDO) achieved the highest efficiency for extracting eight target compounds from the HE leaves. In addition, the key interactions among NADES components were confirmed by Fourier-transform infrared (FT-IR) spectroscopy, providing valuable insights into the extraction mechanism. The UAE process was systematically optimized through single-factor experiments. Subsequently, response surface methodology (RSM) identified the optimal conditions as ultrasonic time of 45 min, solid/liquid ratio of 1:54 g/mL, and ultrasonic temperature of 42 °C. Scanning electron microscopy (SEM) elucidated the microstructural alterations in plant cell walls induced by NADES-UAE, alongside the enhanced penetration and disruption mechanisms. In vitro bioactivity revealed that the NADES-extracted HE exerted strong inhibitory effect on HT-29 colon cancer cells. Overall, these findings demonstrate the high effectiveness and sustainability of NADES-UAE for extracting HE bioactive compounds and provide valuable implications for the industrial-scale production of plant-based functional products.