Metabolism of mono-(2-ethylhexyl) phthalate in Arabidopsis thaliana: Exploration of metabolic pathways by deuterium labeling

Mono-(2-ethylhexyl) phthalate (MEHP) is the primary monoester transformation product of the commonly used plasticizer, di-2-ethylhexyl phthalate (DEHP), and has been frequently detected in various environmental compartments (e.g., soil, biosolids, plants). Plants growing in contaminated soils can take up MEHP, and consumption of the contaminated plants may result in unintended exposure for humans and other organisms. The metabolism of MEHP in plants is poorly understood, but critical for evaluating the potential human and environmental health risks. The present study represents the first attempt to explore the metabolic fate of MEHP in plants. We used Arabidopsis thaliana cells as a plant model and explored metabolic pathways of MEHP using deuterium stable isotope labelling (SIL) coupled with time-of-flight high resolution mass spectrometer (TOF-HRMS). A. thaliana rapidly took up MEHP from the culture medium and mediated extensive metabolism of MEHP. Combining SIL with TOF-HRMS analysis was proved as a powerful method for identification of unknown MEHP metabolites. Four phase Ⅰ and three phase Ⅱ metabolites were confirmed or tentatively identified. Based on the detected transformation products, hydroxylation, oxidation, and malonylation are proposed as the potential MEHP metabolism pathways. In cells, the maximum fraction of each transformation product accounted for 2.8–56.5% of the total amount of metabolites during the incubation. For individual metabolites, up to 2.9–100% was found in the culture medium, suggesting plant excretion. The results in the cell culture experiments were further confirmed in cabbage and A. thaliana seedlings. The findings suggest active metabolism of MEHP in plants and highlight the need to include metabolites in refining environmental risk assessment of plasticizers in the agro-food systems.