Considering the scarce number of studies investigating the oxidative potential of PM2.5 in Italy, a measurement campaign was conducted from February 2020 to October 2020 in a coastal semi-rural site of Basilicata (Southern Italy) with the goal to characterize the fine fraction of ambient particulate matter (PM) and investigate its chemical and toxicological properties, by means of oxidative potential. Different instruments such as an automatic low-volume sampler, an aethalometer and an optical particle counter, were employed for the measurement of PM2.5 mass concentration, equivalent black carbon (eBC) concentration and absorption Ångström exponent (AAE), and particle number size distribution in 0.25–31 μm size range, respectively. 108 daily PM samples, collected on quartz fibre filters, were chemically analysed. Organic (OC) and elemental (EC) carbon content was estimated by thermo-optical transmittance technique (TOT), the concentrations of the main water-soluble ions and total elements were determined by ion chromatography and ED-XRF technique, while the oxidative potential of the water-soluble fraction was estimated through the dithiothreitol (DTT) assay. The mean value of PM2.5 mass concentration was 9.2 ± 2.5 μg/m3 and the average contribution of measured species on PM2.5 mass was 3.3% EC; 19.3% OC; 27.0% secondary inorganic aerosol (sum of SO42−, NH4+ and NO3−), and 10.2% of the other ions and elements. The OC and EC contributions to PM2.5 mass and their mean ratio (OC/EC = 6.6 ± 3.1) suggest that the site is affected by the combined contribution of traffic emissions and biomass combustion (domestic heating and agricultural activities), with the latter prevailing over the traffic, as supported by the mean AAE value of 1.3. The mean OP normalized by sampled volume, OPDTTV, was as high as 0.34 ± 0.22 nmol/min·m3, a value comparable to those recorded for PM2.5 in suburban areas of Italy. The correlation between OPDTTV and the chemical species observed in the PM2.5 samples showed a good agreement with the carbonaceous component OC (r = 0.62) and with some ions, K+ and SO42− (r = 0.60). These results identify combustion sources as the most responsible for the relatively high OPDTTV activity of PM2.5 recorded in this area.