Identification of four novel flavonoid adducts in Arabidopsis thaliana (L.) exposed to isobutyl S-2-diethylaminoethyl methylphosphonothiolate as potential plant exposure biomarkers

Abstract

As vegetation is part of our lives, plants are good candidates as indicators of toxic chemicals. Numerous components in plants may react with toxic chemicals to produce exposure biomarkers. Plant biomarkers formed by the modification of endogenous plant components by chemical warfare agents have not been reported. In this article, the model plant Arabidopsis thaliana (L.) was exposed to the nerve agent isobutyl S-2-diethylaminoethyl methylphosphonothiolate (iBuVX). Some characteristic ions were identified by liquid chromatography–high resolution mass spectrometry and their product ion mass spectra were recorded and interpreted. Some interesting fragmentation pathways were revealed including neutral loss of glucoside, rhamnose and isobutylene. Isobutyl methylphosphonyl modified flavonoids were deduced from assignment of product ions. The element components and the accurate mass of the product ions matched well with those of the proposed fragmentation pathways. The binding site of the nerve agent on flavonoids was proved to be the hydroxyl group on the benzene ring of the flavonoids by density functional theory computation and by the synthesis of the reference chemical, which was confirmed by ¹H–³¹P HMBC NMR. The phosphonyl-modified flavonoids were evaluated for specificity in different plants. Four new flavonoid adducts as potential biomarkers were identified in the leaves of the iBuVX-exposed plant, which provided a novel strategy for the retrospective analysis of organophosphorus exposure for chemical weapon verification and forensic analysis.