The quest for metabolic biomarkers of agrochemicals exposure via in vitro studies and suspect screening

Numerous agrochemicals, including pesticides and herbicides, are applied in modern agriculture, resulting in concerns for the ecosystem and human safety as humans are easily exposed to these compounds. Many agrochemicals, and their transformation products or metabolites, have shown toxicity in in vitro and in vivo studies. However, given the rapid development of novel agrochemicals, for many there is no information about their effects nor about metabolic transformations when ingested by humans. Tracing biomarkers may be the best method for assessing the impacts of agrochemicals. A combination of in vitro metabolism study and suspect screening of human samples (e.g., urine, blood) can be utilized to efficiently find biomarkers for agrochemical exposure. In the work reported here, we determined the in vitro metabolic profiling of six prioritized pesticides and synergists, namely boscalid, carbendazim, piperonyl butoxide, spiroxamine, dimethomorph and fludioxonil, in human liver microsomes. 17 major metabolites were structurally elucidated by high resolution mass spectrometry (HRMS). Major metabolic transformation processes (e.g., hydroxylation, demethylation and oxidation) were proposed for each pesticide. Individual in silico toxicity assessments showed that some metabolites had the same or even enhanced toxicity compared to parent compounds. Information about these metabolites obtained from HRMS was used for suspect screening in human activities related samples. Carbendazim and a metabolite of fludioxonil were identified in wastewater and laboratory urine samples, respectively. Our findings provide concrete evidence for the use of in vitro metabolites as biomarkers in biomonitoring studies of potential exposure to toxic chemicals.