Phenolic Profile and Antioxidant Activity of Chocolates Supplemented with Bioactive Ingredients

The growing demand for functional foods has stimulated the development of chocolate matrices enriched with bioactive ingredients. This study aimed to evaluate the effect of extraction conditions and formulation strategies on the phenolic profile and antioxidant activity of dark chocolate. Five formulations were evaluated: control chocolate (C), chocolate containing vitamin microcapsules (T1), chocolate with DHA/EPA microcapsules (T2), lipid-modified chocolate with structuring oil (T3), and chocolate combining microcapsules with lipid modification (T4). Phenolic compounds were extracted using hydro-organic solvents of different polarities (50% ethanol, 70% methanol, and 70% acetone). Among the tested solvents, 70% methanol showed the highest extraction efficiency, enabling broader detection of phenolic compounds and alkaloids. HPLC-DAD analysis revealed compounds characteristic of cocoa matrices, including epicatechin, gallic acid, vanillin, and procyanidins, as well as the methylxanthines theobromine and caffeine. Among the formulations, T4 exhibited a greater abundance of extractable compounds and the most complex chromatographic profile. Antioxidant activity was evaluated using DPPH radical scavenging and β-carotene/linoleic acid bleaching assays. T4 also showed the highest antioxidant performance in both assays. These findings suggest that the combination of microencapsulation and lipid phase modification may enhance the extractability and functional expression of bioactive compounds, supporting the development of functional chocolate products with added value.