A frequently used analysis system to quantify reactive oxygen species (ROS) in aerosol particles was characterized in detail. The kinetics of different ROS were followed, using fluorescence spectroscopy, upon introduction to the 2-7 dichlorofluorescein (DCFH) fluorescence probe in order to determine the assay’s reactivity. The assay was found to be specific to H2O2, measuring the conversion of ROS to H2O2 in an aqueous system. This implies that the assay is a valid tool for determining sum oxidative stress in an aqueous lung environment.