Deoxynivalenol (DON) sulfonates as major DON metabolites in rats: from identification to biomarker method development, validation and application.

Abstract

Deoxynivalenol (DON) is a trichothecene mycotoxin regularly occurring in cereals. Rats are often used to study toxicokinetics of DON and related compounds, yet only about 30% of the administered dose is typically recovered. Recently, it was reported that DON is partly metabolised to previously undetected DON- and deepoxy-DON (DOM) sulfonate in rats and tentative structures were proposed. The present work describes the production and characterisation of DON-, DOM- and DON-3-glucoside (D3G) sulfonates of three different series; the development and validation of liquid chromatography tandem mass spectrometry (LC-MS/MS)-based methods for determination of DON, DOM, D3G and their sulfonates in rat faeces and urine; and application of the methods to samples from a DON and D3G feeding trial with rats. In addition to previously produced DON sulfonates (DONS) 1, 2 and 3, D3G sulfonates 1, 2 and 3; and DOM sulfonates (DOMS) 2 and 3 were synthesised, purified and characterised. The developed methods showed apparent recoveries of all investigated compounds between 68 and 151% in faeces and between 48 and 113% in urine. The recovery of DON, D3G and their metabolites from faeces and urine of rats (n = 6) administeredin a single dose of 2.0mg/kg b.w. DON or the equimolar amount of D3G was 75 ± 9% for the DON group and 68 ± 8% for the D3G group. DON-, DOM- and D3G sulfonates excreted in faeces accounted for 48 and 47% of the total amount of administered DON and D3G. Urinary excretion of sulfonates was <1%. In both treatment groups, DONS 2 was the major metabolite 0-24h after treatment, whereas DOMS 2 was predominant thereafter. The developed methods can also be used for investigation of DON (conjugate) sulfonate formation in other animal species.