Advertisement

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
Article

Abstract

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.

Keywords

Sorghum Deoxynivalenol Mycotoxin GC-MS Stable isotope Trichothecene 

Abbreviations

DON

Deoxynivalenol

SIDA

Stable isotope dilution assay

GC-MS

Gas chromatography-mass spectrometry

Notes

Acknowledgements

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

Funding

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.

References

  1. Abia WA, Warth B, Sulyok M, Krska R, Tchana AN, Njobeh PB, Dutton MF, Moundipa PF (2013) Determination of Multi-Mycotoxin Occurrence in Cereals, Nuts and Their Products in Cameroon by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). Food Control 31(2):438–453CrossRefGoogle Scholar
  2. Alshannaq A, Yu J-H (2017) Occurrence, Toxicity, and Analysis of Major Mycotoxins in Food. Int J Environ Res Public Health 14(6):632CrossRefGoogle Scholar
  3. Awika JM, Rooney LW (2004) Sorghum Phytochemicals and Their Potential Impact on Human Health. Phytochemistry 65(9):1199–1221CrossRefGoogle Scholar
  4. Ayalew A, Fehrmann H, Lepschy J, Beck R, Abate D (2006) Natural Occurrence of Mycotoxins in Staple Cereals from Ethiopia. Mycopathologia 162(1):57–63.  https://doi.org/10.1007/s11046-006-0027-8 CrossRefGoogle Scholar
  5. De Boevre M, Di Mavungu JD, Maene P, Audenaert K, Deforce D, Haesaert G, Eeckhout M, Callebaut A, Berthiller F, Van Peteghem C (2012) Development and Validation of an LC-MS/MS Method for the Simultaneous Determination of Deoxynivalenol, Zearalenone, T-2-Toxin and Some Masked Metabolites in Different Cereals and Cereal-Derived Food. Food Addit Contam, Part A 29(5):819–835CrossRefGoogle Scholar
  6. Bretz M, Beyer M, Cramer B, Humpf H-U (2006) Stable Isotope Dilution Analysis of the Fusarium Mycotoxins Deoxynivalenol and 3-acetyldeoxynivalenol. Mol Nutr Food Res 50(3):251–260CrossRefGoogle Scholar
  7. Casa AM, Pressoir G, Brown PJ, Mitchell SE, Rooney WL, Tuinstra MR, Franks CD, Kresovich S (2008) Community Resources and Strategies for Association Mapping in Sorghum. Crop Sci 48(1):30–40CrossRefGoogle Scholar
  8. Dykes L (2019) Sorghum Phytochemicals and Their Potential Impact on Human Health. In: Sorghum Methods and Protocols, Humana Press, 1931:121-140.  https://doi.org/10.1007/978-1-4939-9039-9_9
  9. Ediage EN, Van Poucke C, De Saeger S (2015) A Multi-Analyte LC–MS/MS Method for the Analysis of 23 Mycotoxins in Different Sorghum Varieties: The Forgotten Sample Matrix. Food Chem 177:397–404CrossRefGoogle Scholar
  10. Goswami RS, Kistler HC (2004) Heading for Disaster: Fusarium graminearum on Cereal Crops. Mol Plant Pathol 5(6):515–525CrossRefGoogle Scholar
  11. Isakeit T, Prom LK, Wheeler M, Puckhaber LS, Liu J (2008) Mycotoxigenic Potential of Ten Fusarium Species Grown on Sorghum and in Vitro. Plant Pathol J 7:183-186Google Scholar
  12. Juan C, Ritieni A, Mañes J (2012) Determination of Trichothecenes and Zearalenones in Grain Cereal, Flour and Bread by Liquid Chromatography Tandem Mass Spectrometry. Food Chem 134(4):2389–2397CrossRefGoogle Scholar
  13. Kokkonen MK, Jestoi MN (2009) A Multi-Compound LC-MS/MS Method for the Screening of Mycotoxins in Grains. Food Anal Methods 2(2):128–140CrossRefGoogle Scholar
  14. McMaster N, Grothe J, Acharya B, Mehl H, Schmale D (2017) Stable Isotope Dilution Analysis for the Accurate Determination of Deoxynivalenol in Sorghum by GC-MS. In Proceedings of 2017 National Fusarium Head Blight Forum Milwaukee, Wisconsin, p 35. https://scabusa.org/pdfs/NFHBF17_Proceedings_PF_Web.pdf (accessed on 10 July, 2019)
  15. Mirocha CJ, Kolaczkowski E, Xie W, Yu H, Jelen H (1998) Analysis of Deoxynivalenol and Its Derivatives (Batch and Single Kernel) Using Gas Chromatography/Mass Spectrometry. J Agric Food Chem 46(4):1414–1418CrossRefGoogle Scholar
  16. Monbaliu S, Van Poucke C, Detavernier C’l, Dumoulin F, Van De Velde M, Schoeters E, Van Dyck S, Averkieva O, Van Peteghem C, De Saeger S (2009) Occurrence of Mycotoxins in Feed as Analyzed by a Multi-Mycotoxin LC-MS/MS Method. J Agric Food Chem 58(1):66–71CrossRefGoogle Scholar
  17. Neuhof T, Ganzauer N, Koch M, Nehls I (2009) A Comparison of Chromatographic Methods for the Determination of Deoxynivalenol in Wheat. Chromatographia 69(11–12):1457–1462CrossRefGoogle Scholar
  18. Oueslati S, Blesa J, Moltó JC, Ghorbel A, Mañes J (2014) Presence of Mycotoxins in Sorghum and Intake Estimation in Tunisia. Food Addit Contam, Part A 31(2):307–318CrossRefGoogle Scholar
  19. Pena GA, Cavaglieri LR, Chulze SN (2019) Fusarium Species and Moniliformin Occurrence in Sorghum Grains Used as Ingredient for Animal Feed in Argentina. J Sci Food Agric 99(1):47–54CrossRefGoogle Scholar
  20. Pestka J (2010) Toxicological Mechanisms and Potential Health Effects of Deoxynivalenol and Nivalenol. World Mycotoxin J 3(4):323–347CrossRefGoogle Scholar
  21. Rasmussen RR, Storm IMLD, Rasmussen PH, Smedsgaard J, Nielsen KF (2010) Multi-Mycotoxin Analysis of Maize Silage by LC-MS/MS. Anal Bioanal Chem 397(2):765–776CrossRefGoogle Scholar
  22. Rychlik M, Asam S (2008) Stable Isotope Dilution Assays in Mycotoxin Analysis. Anal Bioanal Chem 390(2):617–628CrossRefGoogle Scholar
  23. Sashidhar RB, Ramakrishna Y, Bhat RV (1992) Moulds and Mycotoxins in Sorghum Stored in Traditional Containers in India. J Stored Prod Res 28(4):257–260CrossRefGoogle Scholar
  24. Schmale D, Munkvold G (2009) Mycotoxins in Crops: A Threat to Human and Domestic Animal Health. The Plant Health Instructor,  https://doi.org/10.1094/PHI-I-2009-0715-01. https://www.apsnet.org/edcenter/disimpactmngmnt/topc/Mycotoxins/Pages/default.aspx (accessed on 10 July, 2019)
  25. Serrano AB, Font G, Ruiz MJ, Ferrer E (2012) Co-Occurrence and Risk Assessment of Mycotoxins in Food and Diet from Mediterranean Area. Food Chem 135(2):423–429CrossRefGoogle Scholar
  26. Sulyok M, Krska R, Schuhmacher R (2010) Application of an LC–MS/MS Based Multi-Mycotoxin Method for the Semi-Quantitative Determination of Mycotoxins Occurring in Different Types of Food Infected by Moulds. Food Chem 119(1):408–416CrossRefGoogle Scholar
  27. Tacke BK, Casper HH (1996) Determination of Deoxynivalenol in Wheat, Barley, and Malt by Column Cleanup and Gas Chromatography with Electron Capture Detection. J AOAC Int 79(2):472–475Google Scholar
  28. US Wheat and Barley Scab Initiative Homepage (n.d.). https://scabusa.org/ (accessed on 10 July, 2019)
  29. Vendl O, Berthiller F, Crews C, Krska R (2009) Simultaneous Determination of Deoxynivalenol, Zearalenone, and Their Major Masked Metabolites in Cereal-Based Food by LC–MS–MS. Anal Bioanal Chem 395(5):1347–1354CrossRefGoogle Scholar

Copyright information

© 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

Personalised recommendations