The present work describes the volatile compounds profile and phytochemical content of Ceratonia siliqua L. Fifty different components have been identified. Among them, three constituents are shared i.e., 2-methlybutanoic acid, methyl hexanoate and limonene by different common carob preparations: pulp decoction (PD), seeds decoction (SD) and Rob, a sweet syrup extracted from the pulp of the carob pod. Each extract exhibits different volatile aromatic emission profiles. The antioxidant activity of the extracts was evaluated using three methods, DPPH, ABTS and FRAP, producing a dose-dependent response. The IC_50, when determined by FRAP, gave the lowest values (0.66 ± 0.01, 0.73 ± 0.05 and 0.55 ± 0.00 mg/mL PD, SD and Rob, respectively). The nociception essay, after intraperitoneal injection of acetic acid in mice, demonstrated that Rob, pulp and seeds decoction extracts showed an efficient inhibition of writhes over time, with persistence over 30 min. The SD decoction revealed the highest efficacy in decreasing the writhing reflex (90.3 ± 1.2%; p < 0.001). Furthermore, the proapoptotic activity of SD against three human cell line, THP-1, MCF-7 and LOVO, evaluated by flow cytometry, showed a significantly stronger proapoptotic activity on colon cancer (LOVO) than on the other cell lines, a phenomenon known as phenotypic selectivity.

Carob syrup, a traditional Mediterranean functional beverage obtained from Ceratonia siliqua (L.) pods, has been historically valued for its nutritional properties but is currently underutilized. This study compared the prebiotic potential of three handmade carob syrups produced by Tunisian women with commercial benchmarks from Italy, Greece and Cyprus. The prebiotic activity was evaluated by prebiotic scores, bifidogenic activity and volatilome characterization (SPME GC/MS) together with physicochemical and nutritional parameters. The results showed that Tunisian handmade products exhibited lower growth of pathogenic Escherichia coli compared to commercial samples. The prebiotic activity, tested against probiotic lactobacilli and bifidobacteria mixtures, showed a hierarchy of efficacy: fructo-oligosaccharides (FOSs) > Tunisian handmade products > Greek and Cypriot benchmarks > Italian benchmark. Volatilome analysis revealed about 40 compounds, mainly organic acids and aldehydes, with higher concentrations in handmade products. Positive correlations were found between prebiotic activity and short-chain fatty acids and n-hexadecanoic acid, while furfural showed negative correlations. The Tunisian artisanal products showed a higher prebiotic potential compared to the commercial counterparts, due to their higher content and diversity of organic acids. However, the presence of furfural in Tunisian products needs to be monitored due to potential toxicity concerns.