Chlorpyrifos (CPF) is a widely used organophosphorus insecticide that tends to form bound residues (BRs) in soils. However, the stability and biological activity of CPF-BRs remain to be explored. Facilitated by carbon-14 tracing, this study obtained CPF-BRs initially formed in two typical paddy soils (¹⁴C-CPF-BR_in), and further investigated their release, transformation and phytoavailability using duckweed (Lemna minor) as a model aquatic organism. Most ¹⁴C-CPF-BR_in in soils were composed of the parent CPF and its metabolite 3,5,6-trichloro-2-pyridinol (2-OH-TCP), which was mainly formed through reversible entrapment by soil fulvic acids and humin (>80%). At 36 d, 66.67-80.90% of the ¹⁴C-CPF-BR_in was released, of which only 2-OH-TCP could be released into the water and absorbed by the duckweed, with bioconcentration factors ranging from 247.99 to 324.68 L kg^-1. The subsequent metabolism of released ¹⁴C-CPF-BR_in in duckweed included phase I metabolism from 2-OH-TCP to 4-OH-TCP and phase II metabolism of conjugation of TCP with plant endogenous amino acids. The study suggested that CPF bound residues have high bioavailability in paddy field environments. Given that many pesticides and non-pesticide chemicals share structures analogous to CPF, the findings have important implications for better understanding the environmental and human health risks of man-made chemicals.

The study of pesticide metabolism in crops is critical for assessing the mode of action and environmental risks of pesticides. However, the study of pesticide metabolism in crops is usually complicated and it is often a daunting challenge to accurately screen the metabolites of novel pesticides in complex matrices. This study demonstrated a combined use of high-specific activity carbon-14 labeling and high-resolution mass spectrometry (HSA-¹⁴C-HRMS) for metabolism profiling of a novel neonicotinoid cycloxaprid in rice. By generating the characteristic radioactive peaks on the liquid chromatogram, the use of ¹⁴C can eliminate the severe interference of complex matrices and quickly probe target compounds; by producing ion pairs with unique abundance ratios on HRMS, high-specific activity labeling can effectively exclude false matrix positives and promote the elucidation of metabolite structure. The structures of 15 metabolites were identified, three of which were further confirmed by authentic standards. Based on these metabolites, a metabolic profile of cycloxaprid was established, which includes denitrification, demethylation, imidazolidine hydroxylation and ring cleavage olefin formation, oxidation and carboxylation reactions. The strategy of combining high-specific activity ¹⁴C labeling and HRMS offers unique advantages and provides a powerful solution for profiling unknown metabolites of novel pesticides in complex matrices, especially when traditional non-labeling methods are not feasible.