Screening of dioxin-like compounds in bio-composts and their materials: chemical analysis and fractionation-directed evaluation of AhR ligand activities using an in vitro bioassay

Abstract

Application of the dioxin responsive-chemical activated luciferase gene expression (DR-CALUX) bioassay to three compost products (kitchen garbage compost, treated sludge compost, and poultry manure compost) and their organic waste materials was performed for the screening of dioxin-like compounds. Phased sample preparation was conducted for the bioassay to yield separate crude, acid-stable, and carbon column fractions. Chemical analyses were also conducted for polychlorinated dibenzo-p-dioxin and dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs), and polyaromatic hydrocarbons (PAHs). Their determined WHO-PCDD/PCDF/dl-PCB-TEQ values were compared to the DR-CALUX results expressed as CALUX-TEQ to ascertain their contributions to overall activity. Overall all three compost products were polluted to a low level by persistent PCDD/PCDF/dl-PCBs in a few pg/g WHO-TEQ concentration range. The organic crude fraction showed non-persistent CALUX-TEQ values 2-4 orders of magnitude higher than those of WHO-TEQ. For the acid-resistant fractions of all samples, persistent CALUX-TEQ values were 1-8-fold higher than WHO-PCDD/PCDF/dl-PCB-TEQ values. Both TEQs correlated well for this fraction, which confirms the availability of CALUX for estimation of the actual WHO-TEQ level of the samples using reflux method with 44% sulfuric acid/silica gel, although the compost and material samples examined in this study contained much lower levels of WHO-TEQ: less than 1 pg/g in almost all cases. Additional carbon column treatment enhanced the potency and efficacy of CALUX activity in the three compost samples, which showed 1.6-4.4 times higher values of CALUX-TEQ than those obtained for the acid-resistant fraction. Although further DR-CALUX data collection for compost samples will be necessary, DR-CALUX is useful as a rapid screening method for monitoring dioxin-like compounds in compost.