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

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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.

Formation of deoxynivalenol (DON)- and deepoxy-DON sulfonates in rats

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Acknowledgments

The authors would like to thank the Federal Ministry of Economy, Family and Youth, the National Foundation for Research, Technology and Development and BIOMIN Holding GmbH for funding the Christian Doppler Laboratory for Mycotoxin Metabolism. In addition, we would like to thank Alexander Frank and Verena Klingenbrunner for providing the crude preparation of DOM and preparing the standard of [13C]-DOM. Furthermore, we are grateful to Dr. Ilse Dohnal for the opportunity to measure on the 5500 Triple Quad system. We would also like to thank Prof. Gerhard Adam for fruitful discussions on mechanisms of DON sulfonate formation in rats. Finally, we thank Melanie Leitner for supplying fresh faeces of Sprague-Dawley rats.

Conflict of interest

The authors declare to have no conflict of interests.

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Authors and Affiliations

  1. 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 E. Schwartz-Zimmermann, Veronika Nagl, Veronika Slavik & Franz Berthiller

  2. Institute of Applied Synthetic Chemistry, Vienna University of Technology, Getreidemarkt 9/163-OC, 1060, Vienna, Austria

    Christian Hametner

  3. BIOMIN Research Center, Technologiezentrum Tulln, Technopark 1, 3430, Tulln, Austria

    Wulf-Dieter Moll

Authors
  1. Heidi E. Schwartz-Zimmermann
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  2. Christian Hametner
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  3. Veronika Nagl
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  4. Veronika Slavik
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  5. Wulf-Dieter Moll
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  6. Franz Berthiller
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Corresponding author

Correspondence to Heidi E. Schwartz-Zimmermann.

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Schwartz-Zimmermann, H.E., Hametner, C., Nagl, V. et al. Deoxynivalenol (DON) sulfonates as major DON metabolites in rats: from identification to biomarker method development, validation and application. Anal Bioanal Chem 406, 7911–7924 (2014). https://doi.org/10.1007/s00216-014-8252-3

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  • DOI: https://doi.org/10.1007/s00216-014-8252-3

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