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

High Resolution Image Download MS PowerPoint Slide Bombyx mori ( B. mori ) 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 B. mori 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 Staphylococcus aureus and Escherichia coli . 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.