A Semi-automated Instrument for Cellular Oxidative Potential Evaluation (SCOPE) of Water-soluble Extracts of Ambient Particulate Matter

Abstract

Abstract. Several automated instruments exist to measure the acellular oxidative potential (OP) of ambient particulate matter (PM). However, cellular OP of the ambient PM is still measured manually, which severely limits the comparison between two types of assays. Cellular assays could provide a more comprehensive assessment of the PM-induced oxidative stress, as they incorporate more biological processes involved in the PM-catalyzed reactive oxygen species (ROS) generation. Considering this need, we developed a first of its kind semi-automated instrument for measuring the cellular OP based on a macrophage ROS assay using rat alveolar macrophages. The instrument named SCOPE – Semi-automated instrument for Cellular Oxidative Potential Evaluation, uses dichlorofluorescein diacetate (DCFH-DA) as a probe to detect the OP of PM samples extracted in water. SCOPE is capable of analyzing a batch of six samples (including one negative and one positive control) in five hours and is equipped to operate continuously for 24-hours with minimal manual intervention after every batch of analysis, i.e., after every five hours. SCOPE has a high analytical precision as assessed from both positive controls and ambient PM samples (CoV < 17 %). The results obtained from the instrument were in good agreement with manual measurements using tert-Butyl hydroperoxide (t-BOOH) as the positive control (slope = 0.83 for automated vs. manual, R2 = 0.99) and ambient samples (slope = 0.83, R2 = 0.71). We further demonstrated the ability of SCOPE to analyze a large number of both ambient and laboratory samples, and developed a dataset on the intrinsic cellular OP of several compounds, such as metals, quinones, PAHs and inorganic salts, commonly known to be present in ambient PM. This dataset is potentially useful in future studies to apportion the contribution of key chemical species in the overall cellular OP of ambient PM.