Effect of Extraction Method on the Bioactive Composition, Antimicrobial Activity and Phytotoxicity of Pomegranate By-Products

Abstract

Pomegranate by-products can be an asset to the food industry due to the richness in bioactive and antimicrobial compounds. This work studied the influence of conventional solvent and sonication-assisted extraction methods on the bioactive profile, antimicrobial properties, and phytotoxicity effect of the peels and seeds extracts from Acco, Big Full, and Wonderful pomegranate cultivars. The bioactive composition of the extracts was evaluated for the content of total phenolics, total flavonoids, and antioxidant activity (expressed as the half-maximal inhibitory concentration-IC₅₀) by spectrophotometric methods, while the tannins were determined by titration and the anthocyanins were estimated by the pH-differential method. For the evaluation of the antimicrobial activity, the disk diffusion method of Kirby-Bauer was adapted through inhibition halos against <i>Escherichia coli</i>, <i>Pseudomonas aeruginosa</i>, <i>Staphylococcus aureus</i>, <i>Bacillus cereus</i>, and <i>Yarrowia lipolytica</i>. The extracts' phytotoxicity was evaluated in vitro on garden-cress seeds. Extracts from conventional extraction were richer in total phenolics, expressed as gallic acid equivalents (0.16-0.73 mg GAE/mg extract), while those from sonication-assisted extraction had higher contents of total flavonoids, expressed as catechin equivalents (0.019-0.068 mg CATE/mg extract); anthocyanins, expressed as cyanidin-3-glucoside (0.06-0.60 µg C3G/mg, dry basis); and antioxidant activity (IC₅₀, 0.01-0.20 mg/mL). All extracts were more effective against Gram-positive bacteria and yeasts than Gram-negative bacteria. In general, the sonication-assisted extracts led to higher inhibition halos (8.7 to 11.4 mm). All extracts presented phytotoxicity against garden-cress seeds in the tested concentrations. Only the lowest concentration (0.003 mg/mL) enabled the germination of seeds and root growth, and the sonication-assisted extracts showed the highest Munoo-Liisa vitality index (51.3%). Overall, sonication-assisted extraction obtained extracts with greater bioactive and antimicrobial potential and less phytotoxicity.