Enhancing DTT assays for reactive oxygen species detection in atmospheric particulate matter: key factors and methodological insights

Abstract

Accurate detection of reactive oxygen species (ROS) in atmospheric particulate matter (PM) is essential for assessing the oxidative potential (OP) of airborne pollutants and their associated health risks. While multiple methods exist for ROS detection, inconsistencies in assay conditions often lead to variable outcomes, limiting cross-study comparability. This review systematically evaluates key methodological factors affecting the dithiothreitol (DTT) assay, one of the most widely used techniques for measuring the OP of PM. Critical parameters—including intrinsic assay variables such as initial DTT concentration and incubation conditions, as well as extrinsic factors such as light exposure and metal–organic interactions—are analyzed to identify sources of variability. To improve sensitivity and reliability, this study proposes standardized protocols, the incorporation of positive controls, and methodological refinements. By addressing these challenges, this review enhances the accuracy of ROS detection and contributes to a more comprehensive understanding of the OP of PM, with significant implications for environmental monitoring and public health.