title: 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) administered in a single dose of 2.0 mg/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 0-24 h 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.
authors.affiliation: Christian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department for Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences, Vienna, Konrad Lorenz Str. 20, 3430, Tulln, Austria, heidi.schwartz@boku.ac.at.
authors.forename: Heidi E
authors.initials: HE
authors.lastname: Schwartz-Zimmermann
authors.forename: Christian
authors.initials: C
authors.lastname: Hametner
authors.forename: Veronika
authors.initials: V
authors.lastname: Nagl
authors.forename: Veronika
authors.initials: V
authors.lastname: Slavik
authors.forename: Wulf-Dieter
authors.initials: WD
authors.lastname: Moll
authors.forename: Franz
authors.initials: F
authors.lastname: Berthiller
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D005243:Feces / Q000737:chemistry / Q000382:microbiology; D057230:Limit of Detection; D008297:Male; D009183:Mycotoxins / Q000032:analysis* / Q000378:metabolism / Q000652:urine; D051381:Rats; D017207:Rats, Sprague-Dawley; D013451:Sulfonic Acids / Q000032:analysis* / Q000378:metabolism / Q000652:urine; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014255:Trichothecenes / Q000032:analysis* / Q000378:metabolism / Q000652:urine
chemical_list: D009183:Mycotoxins; D013451:Sulfonic Acids; D014255:Trichothecenes; C007262:deoxynivalenol