Quantification of the Mycotoxin Deoxynivalenol (DON) in Sorghum Using GC-MS and a Stable Isotope Dilution Assay (SIDA)

  • Nicole McMaster
  • Bhupendra Acharya
  • Kim Harich
  • Jan Grothe
  • Hillary L. Mehl
  • David G. SchmaleIIIEmail author


Sorghum has gained popularity with consumers as a grain source with its gluten-free and high protein dietary characteristics. Acreage has increased recently, in part due to the demand for an alternative feed source for poultry and swine. New information is needed about the level of mycotoxin contamination in sorghum accessions, and accurate and affordable analytical methods need to be developed and tested to quantify mycotoxins in sorghum. A traditional method (solid-phase extraction chromatography with C18, followed by GC-MS) to quantify the mycotoxin deoxynivalenol (DON) produced inconsistent DON values following controlled spiking and recovery experiments with different sorghum accessions. Consequently, we incorporated a stable isotope (d1-DON) as an internal standard into a traditional GC-MS method. This method, stable isotope dilution assay (SIDA), was used to accurately determine DON levels in 196 sorghum samples representing 98 different accessions. Of the 98 accessions tested (two samples per accession), 76 of the accessions had DON levels that were greater than the limit of detection for both methods (0.20 μg g−1). For a USA regulatory threshold of 1 μg g−1, about one third of all the accessions (26/76) had at least 20% more DON using the SIDA method. For a regulatory threshold of 5 μg g−1, about 7% of all the accessions (5/76) had at least 20% more DON using the SIDA method. Using SIDA, the amount of DON in a sorghum sample can be accurately and reliably quantitated by basing calculations on the recovery of d1-DON, and this method may have future applications for quantifying DON from samples with complex matrices.


Sorghum Deoxynivalenol Mycotoxin GC-MS Stable isotope Trichothecene 





Stable isotope dilution assay


Gas chromatography-mass spectrometry



We thank Erica Pack, Hamilton Crockett, and James Benson for their support and assistance with this work.


This work was supported in part by grants to D. Schmale from the Virginia Small Grains Board and the United States Wheat and Barley Scab Initiative (USWBSI), and by grants to H. Mehl from Smithfield Murphy Brown. This material is based upon work supported by the U.S. Department of Agriculture, under Agreement No. 58-0206-6-017. Any opinions, findings, conclusions, or recommendationsexpressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture. The conclusions presented here are those of the authors and do not necessarily reflect the views of the sponsors.

Compliance with Ethical Standards

Conflict of Interest

Authors N. McMaster, B. Acharya, K. Harich, J. Grothe, H. L. Mehl, and D. Schmale declare that they have no conflicts of interest.

Ethical Approval

This article does not contain any studies with human participants performed by any of the authors.

Informed Consent

Not applicable.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Plant and Environmental SciencesVirginia TechBlacksburgUSA
  2. 2.Virginia Tech Tidewater Agricultural Research and Extension CenterSuffolkUSA
  3. 3.Department of BiochemistryVirginia TechBlacksburgUSA

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