Sequential green extraction, identification, and encapsulation of bioactive compound from Phellinus linteus fruiting body

Phellinus linteus, a traditional medicinal mushroom, is highly valued for its health benefits. The commercially available fruiting body of Phellinus linteus (PL) was extracted and characterized to elucidate its potential nutritional bioactivity. A sequential extraction method was employed, starting with conventional ethanol extraction (PL-E), followed by mechanical ball-mill assisted ethanol extraction (PL-B) or supercritical CO₂ cosolvent (PL-C), resulting in the identification of 19, 9, and 10 tentative compounds, respectively. The extracts comprised non-polar compounds, including triterpenoids and sterols in PL-E, and polyphenols and long-chain fatty acids in PL-B and PL-C. Hispidin and hispolon were found exclusively in PL-E. The PL-E demonstrated anticancer potential against MCF-7 cells, while the PL-C showed the highest activity against MCF-7 and HT-29 cells, suggesting that this high bioactivity fraction may be a promising candidate for anticancer studies. The PL-E, which exhibited high yield and bioactivity with various bioactive compounds, underwent complexation encapsulation with β-cyclodextrin. This study also proposes a potential green method to enhance these compounds' bioactivity value and bioaccessibility, supporting their development for food, nutraceutical, and pharmacological applications.