Atmospheric particulate matter (PM) has been related to numerous adverse health effects in humans. Nowadays, it is believed that one of the possible mechanisms of toxicity could be the oxidative stress, which involves the development of reactive oxygen species (ROS). Different assays have been proposed to characterize oxidative stress, such as dithiothreitol (DTT) and ascorbic acid (AA) acellular assays (OPDTT and OPAA), as a metric more relevant than PM mass measurement for PM toxicity. This study evaluates the OP of the bioaccessible fraction of 65 PM10 samples collected at an Atlantic Coastal European urban site using DTT and AA assays. A physiologically based extraction (PBET) using Gamble’s solution (GS) as a simulated lung fluid (SLF) was used for the assessment of the bioaccessible fraction of PM10. The use of the bioaccessible fraction, instead of the fraction assessed using conventional phosphate buffer and ultrasounds assisted extraction (UAE), was compared for OP assessment. Correlations between OPDTT and OPAA, as well as total and bioaccessible concentrations of polycyclic aromatic hydrocarbons (PAHs) and metal(oid)s, were investigated to explore the association between those compounds and OP. A correlation was found between both OP (OPDTT and OPAA) and total and bioaccessible concentrations of PAHs and several metal(oid)s such as As, Bi, Cd, Cu, Ni, and V. Additionally, OPDTT was found to be related to the level of K+.