<i>Bombyx mori</i> Pupae as a Novel Ingredient from an Underutilized Sericulture Product: A Dual Approach Based on Sustainable Extractions and Sample Pretreatment Strategies

<i>Bombyx mori</i> (<i>B. mori</i>) pupae, a major byproduct of the silk industry, can be upcycled as a source of bioactive lipids for food, nutraceutical, and cosmetic applications. This study aimed to sustainably recover the lipophilic fraction from<i>B. mori</i> using supercritical fluid extraction (SFE) assisted by natural hydrophobic solvents (SFE-NHSolv) as an alternative to Soxhlet (SOX) extraction with hexane while also evaluating the impact of two different pretreatment methods, freeze-drying (FD) and blanching (B), followed by FD (B+FD). The process yield, lipid and carotenoid profiles, bioactivities (antioxidant and antibacterial), and environmental performance of SOX and SFE-NHSolv were assessed. Among the processes, SFE-NHSolv with B+FD achieved the highest yield (55.98%) compared with Soxhlet (38.3%). The fatty acid profile revealed that the lipid fraction is mainly composed of linolenic, oleic, and palmitic acids. Five carotenoids were tentatively identified in SOX extracts including lutein, zeaxanthin, and β-carotene. Extracts exhibited higher antioxidant capacity by ABTS (3.25-15.44 μmol TE g^-1) than by DPPH (2.59-7.23 μmol TE g^-1), with SFE-NHSolv B+FD showing the highest value. No antibacterial activity was observed against <i>Staphylococcus aureus</i> and<i>Escherichia coli</i>. The sustainability assessment confirmed SFE-NHSolv as a cleaner and more eco-efficient process than SOX, capable of producing ready-to-use extracts. In summary, this study contributes to the advancement of sustainable extraction technologies. It promotes the sustainable use of silk industry byproducts, offering eco-friendly solutions for developing high-value ingredients for the food, nutraceutical, and cosmetic sectors.