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. It is being suggested that particle-bound or particle-inducedreactive oxygen species (ROS), which significantly contribute to theoxidative potential (OP) of aerosol particles, are a promising metriclinking aerosol compositions to toxicity and adverse health effects.However, accurate ROS quantification remains challenging due to the reactiveand short-lived nature of many ROS components and the lack of appropriateanalytical methods for a reliable quantification. Consequently, it remainsdifficult to gauge their impact on human health, especially to identify howaerosol particle sources and atmospheric processes drive particle-bound ROSformation in a real-world urban environment. In this study, using a novel online particle-bound ROS instrument (OPROSI),we comprehensively characterized and compared the formation of ROS insecondary organic aerosols (SOAs) generated from organic compounds thatrepresent anthropogenic (naphthalene, SOANAP) and biogenic (β-pinene, SOAβPIN) precursors. The SOA mass was condensed ontosoot particles (SP) under varied atmospherically relevant conditions(photochemical aging and humidity) to mimic the SOA formation from a mixingof traffic-related carbonaceous primary aerosols and volatile organic compounds (VOCs). We systematicallyanalyzed the ability of the aqueous extracts of the two aerosol types(SOANAP-SP and SOAβPIN-SP) to induce ROS productionand OP. We further investigated cytotoxicity and cellular ROS productionafter exposing human lung epithelial cell cultures (A549) to extracts of thetwo aerosols. A significant finding of this study is that more than 90 %of all ROS components in both SOA types have a short lifetime, highlightingthe need to develop online instruments for a meaningful quantification ofROS. Our results also show that photochemical aging promotes particle-boundROS production and enhances the OP of the aerosols. Compared to SOAβPIN-SP, SOANAP-SP elicited a higher acellular and cellular ROSproduction, a higher OP, and a lower cell viability. These consistent resultsbetween chemical-based and biological-based analyses indicate thatparticle-bound ROS quantification could be a feasible metric to predictaerosol particle toxicity and adverse human effects. Moreover, the cellularROS production caused by SOA exposure not only depends on aerosol type butis also affected by exposure dose, highlighting a need to mimic the processof particle deposition onto lung cells and their interactions asrealistically as possible to avoid unknown biases.

Are reactive oxygen species (ROS) a suitable metric to predict toxicity of carbonaceous aerosol particles?