From Solvent Design to Biological Response: Structure-Property Relationships of Edible Natural Deep Eutectic Solvents for the Extraction of Nigella sativa Bioactives.

Edible Natural Deep Eutectic Solvents (NADES) offer a route to ready-to-use extracts without solvent removal. This study examined how rational formulation design influences physicochemical properties, extraction performance, energy efficiency, thermal behavior, and cytocompatibility during bioactive recovery from Nigella sativa seeds. Twelve formulations spanning malic acid-polyol, citric acid-polyol, binary polyol, and ternary acid-polyol families were evaluated under standardized ultrasound-assisted conditions and compared with ethanolic ultrasound-assisted extraction, Soxhlet extraction, and cold-pressed oils. Selected NADES formulations outperformed the conventional systems. E1 (malic acid:glycerol:water) achieved the highest total phenolic content and lowest specific energy consumption (40.00 kJ mg-1 GAE), below the Soxhlet benchmark (55.17 kJ mg-1 GAE), whereas E9 (glycerol:xylitol:water) exhibited the greatest ABTS activity. Neat-NADES apparent pH and viscosity were inversely associated with total phenolic content, while viscosity and density were inversely associated with ABTS activity. ATR-FTIR showed stable, solvent-dominated fingerprints over 15 days, whereas DSC better differentiated neat NADES from their extracts. Most systems remained cytocompatible at 10,000× dilution, while acid-polyol formulations reduced viability at 500×; partial neutralization of N5/E5 restored viability. NADES performance was formulation- and application-dependent, requiring joint optimization of composition, acidity, viscosity, energy efficiency, and biologically compatible concentration.