2.1.3. Chemical Analysis of Pomegranate Extracts

The chemical analyses were performed on PFE and PPE after freeze drying. The identification and quantification of the various phenolic components was determined using HPLC-DAD as described by Romani et al. [34]. Analyses were carried out using a Varian ProStar 210 multisolvent pump and ProStar 335 photodiode array detector (Varian BV, Middelburg, The Netherlands). A Phenomenex Kinetex Phenyl-Hexyl 100 A 150 × 4.6 mm reverse-phase C18 column (Danaher Corporation, Torrance, CA, USA), with the same pre-column, was used for the separation and analysis at 25 °C. Water/acetic acid (99.9:0.1) and methanol/water CH₃COOH made up the eluent (95:4.9:0.1). In particular, α and β punicalagin, and ellagic acid were identified by comparing the retention time and the spectrum with HPLC-grade standards (Sigma-Aldrich, St. Louis, MO, USA). The quantification was obtained using the calibration curve made with the relative standard. All analyses were performed in triplicate.