Response Surface Modeling and Optimization of the Extraction of Phenolic Antioxidants from Olive Mill Pomace

Bioactive compounds from olive mill pomace (OMP) were extracted through a two-step solid-liquid extraction procedure considering four factors at five levels of a central composite rotatable response surface design. The influence of the process variables time of the primary extraction (2.0–4.0 h), solvent-to-sample ratio during the primary extraction (5.0–10.0 mL/g), time of the secondary extraction (1.0–2.0 h), and the solvent-to-sample ratio during the secondary extraction (3.0–5.0 mL/g) were examined. The content of bioactive compounds was determined spectrophotometrically, and the individual phenolic compounds were evaluated by reserved-phase high-performance liquid chromatography (RP-HPLC). The Derringer’s function was used to optimize the extraction process, and the best conditions were found to be 3.2 h for the primary extraction, 10.0 mL/g for the solvent-to-sample ratio and 1.3 h for the secondary extraction associated with a solvent-to-sample ratio of 3.0 mL/g, obtaining a total phenolic content of 50.0 (expressed as mg gallic acid equivalents (GAE)/g dry weight (dw). The response surface methodology proved to be a great alternative for reducing the number of tests, allowing the optimization of the extraction of phenolic antioxidants from OMP with a reduced number of experiments, promoting reductions in cost and analysis time.     

Olive Pomace Phenolic Compounds Stability and Safety Evaluation: From Raw Material to Future Ophthalmic Applications

Nowadays, increasing interest in olive pomace (OP) valorization aims to improve olive’s industry sustainability. Interestingly, several studies propose a high-value application for OP extracts containing its main phenolic compounds, hydroxytyrosol and oleuropein, as therapy for ocular surface diseases. In this work, the stability and accessibility of OP total phenolic and flavonoid content, main representative compounds, and antioxidant activity were assessed under different pretreatment conditions. Among them, lyophilization and supercritical CO₂ extraction were found to increase significantly most responses measured in the produced extracts. Two selected extracts (CONV and OPT3) were obtained by different techniques (conventional and pressurized liquid extraction); Their aqueous solutions were characterized by HPLC-DAD-MS/MS. Additionally, their safety and stability were evaluated according to EMA requirements towards their approval as ophthalmic products: their genotoxic effect on ocular surface cells and their 6-months storage stability at 4 different temperature/moisture conditions (CPMP/ICH/2736/99), together with pure hydroxytyrosol and oleuropein solutions. The concentration of hydroxytyrosol and oleuropein in pure or extract solutions was tracked, and possible degradation products were putatively identified by HPLC-DAD-MS/MS. Hydroxytyrosol and oleuropein had different stability as standard or extract solutions, with oleuropein also showing different degradation profile. All compounds/extracts were safe for ophthalmic use at the concentrations tested.     

One-Step Extraction of Olive Phenols from Aqueous Solution Using β-Cyclodextrin in the Solid State,a Simple Eco-Friendly Method Providing Photochemical Stability to the Extracts

The extraction of phenolic compounds from olive mill wastes is important, not only to avoid environmental damages, but also because of the intrinsic value of those biophenols, well-known for their high antioxidant potential and health benefits. This study focuses on tyrosol (Tyr) and hydroxytyrosol (HT), two of the main phenolic compounds found in olive mill wastes. A new, simple, and eco-friendly extraction process for the removal of phenolic compounds from aqueous solutions using native β-cyclodextrin (β-CD) in the solid state has been developed. Several β-CD/biophenol molar ratios and biophenol concentrations were investigated, in order to maintain β-CD mostly in the solid state while optimizing the extraction yield and the loading capacity of the sorbent. The extraction efficiencies of Tyr and HT were up to 61%, with a total solid recovery higher than 90% using an initial concentration of 100 mM biophenol and 10 molar equivalents of β-CD. The photochemical stability of the complexes thus obtained was estimated from ∆E*ab curve vs. illumination time. The results obtained showed that the phenols encapsulated into solid β-CD are protected against photodegradation. The powder obtained could be directly developed as a safe-grade food supplement. This simple eco-friendly process could be used for extracting valuable biophenols from olive mill wastewater.