Development and Characterization of Calcium Ion-Enhanced Nanophytosomes Encapsulating Pomegranate Fruit Extract

Nanophytosomes (NP_S) loaded with whole pomegranate fruit extract with peel and arils (PFE) at different levels of phosphatidylcholine (PC) were produced using a thin-film hydration method and reinforced with calcium ions. PFE was obtained by pressing whole pomegranates, followed by mixing with PC at ratios of 1:1, 1:2, and 1:3, which then strengthens the phytosome wall by CaCl₂ solutions (1.35 and 2.70 mM) and lyophilized to create a stable powder form. The characteristics of the NP powders, including encapsulation efficiency (EE), particle size, ζ-potential, polydispersity index (PDI), structure, microstructure, and thermal properties, were evaluated. Additionally, the storage stability of phenolic compounds over two months was investigated. The PFE powder demonstrated appropriate characteristics for incorporation into the phytosome system, with a total phenol content of 371.19 mg GAE/g dry weight, anthocyanins at 300.68 mg/g, flavonoids at 194 mg/100 g, and an antioxidant activity of 90.98%. The highest EE was determined to be 98.53%, indicating its unique ability as a nano-carrier. PFE-loaded NPs showed favorable characteristics, such as low PDI values (< 0.5), smaller particle size (170 nm), and a spherical morphology. The PFE-NP had a particle size of 128.6 nm, zeta potential of -40.15 mV, mobility of -3.15 μm cm/Vs, PDI of 0.168, and EE of 98.53%. The optimized nanoparticles remained stable for two months at 4°C, with negligible changes in particle size (~10 nm), total phenol content (TPC), and PDI of the PFE-Nanophytosomes. All NP samples showed better stability at storage temperatures over 60 days. PEF-NPs improved the stability of phenolic compounds while improving solubility, masking taste, and delivery to target tissues, which can be considered in future applications.