Development and application of a novel bioassay system for dioxin determination and AhR activation evaluation in ambient-air samples.

Airborne persistent toxic substances are associated with health impacts resulting from air pollution, for example, dioxins, dioxin like polychlorinated biphenyls, and certain polycyclic aromatic hydrocarbons (PAHs), which activate aryl hydrocarbon receptors (AhR) and thereby produce adverse outcomes. Thus, a bioassay for evaluating AhR activation is required for risk assessment of ambient-air samples, and for this purpose, we developed a new and sensitive recombinant mouse hepatoma cell line, CBG2.8D, in which a novel luciferase-reporter plasmid containing two copies of a newly designed dioxin-responsive domain and a minimal promoter derived from a native gene were integrated. The minimal detection limit for 2,3,7,8-tetrachlorodibenzo-p-dioxin with this assay system was 0.1 pM. We used CBG2.8D to determine dioxin levels in 45 ambient-air samples collected in Beijing. The measured bioanalytical equivalent (BEQ) values were closely correlated with the toxic equivalent values obtained from chemical analysis. In haze ambient-air samples, the total activation of aryl hydrocarbon receptors (TAA) was considerably higher than the BEQof dioxin-rich fractions, according to the results of the cell-based bioassay. Notably, the haze samples contained abundant amounts of PAHs, whose relative toxicity equivalent was correlated with the TAA; this finding suggests that PAHs critically contribute to the AhR-related biological impacts of haze ambient-air samples.