BIOACTIVE CHARACTERIZATION OF BERGAMOT LEAF EXTRACT (CITRUS BERGAMIA) AND ITS POTENTIAL HEALTH APPLICATIONS

The leaves of bergamot (Citrus bergamia), often discarded, are a source of bioactive compounds. This study aimed to assess the bioactive compounds, antioxidant capacity, and potential health applications of bergamot leaf extract (BLE). Bergamot leaves from Calabria, Italy, were extracted using a water/ethanol solution (30/70%, v/v), filtered, eluted with pure ethanol, vacuum distilled at 40 °C, and dried using spray drying, yielding the BLE. The total phenolic compounds (TPC), flavonoids (TF), carotenoids, and antioxidant capacity (DPPH, FRAP, ABTS) of BLE were determined via UV-Vis spectrophotometry. Biogenic amine (BA) analysis (µg/g) was performed using HPLC, and metabolomic analysis (µM/mg) was evaluated via 1H-NMR. The impact of BLE on cell viability was assessed using the MTT assay in LPS-induced 3T3-L1 cells after 24-72 hours. The IC50 of BLE was determined in BALB/c 3T3 A31 cells after 48 hours via red crystal staining. Acute oral toxicity (LD50) was estimated, and biochemical pathway activation or inhibition, as well as potential adverse effects, were predicted using Way2DRUG software. BLE exhibited low levels of carotenoids (0.5±0.1), but high TPC (102±8) and TF (47±3), along with significant antioxidant capacity (DPPH: 35±1, FRAP: 19±0.1, ABTS: 10±1). The BA profile included tryptamine, agmatine, spermidine, and seven others. Metabolomic analysis identified 79 compounds, with the most abundant erythritol, malate, and glycerol. Optimal dosages for enhancing cell viability were 1-10 µg/mL. BLE's IC50 was 1.042 mg/mL, and its estimated LD50 value was 1401.67 mg/Kg (112.13 g for an 80 Kg human). BLE's primary therapeutic potentials include mucositis treatment, antimigraine effects, 5-HT agonists for neurological disorders, macrophage stimulation, and vasoprotective properties. However, potential adverse effects like ocular irritation, enzyme alterations, and hepatotoxicity were also noted. In conclusion, BLE is a promising source of bioactive compounds with high antioxidant capacity, offering potential applications in treatments for mucositis, migraines, anxiety, immunotherapy, and cardiovascular health.