Separation of trichothecene mycotoxins in a matrix of hemp (Cannabis sativa L.) seed, and quantification of deoxynivalenol in hemp grain infected with Fusarium graminearum

Authors: Kagan, Isabelle; RICCIARDI, MAGDALENA - University Of Kentucky; SMITH, HENRY - University Of Kentucky; SCHENDEL, RACHEL - University Of Kentucky; PEARCE, ROBERT - University Of Kentucky; GAUTHIER, NICOLE - University Of Kentucky

Submitted to: Journal of the Science of Food and Agriculture
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/29/2026
Publication Date: 5/20/2026
Citation: Kagan, I., Ricciardi, M., Smith, H.S., Schendel, R.R., Pearce, R.C., Gauthier, N. 2026. Separation of trichothecene mycotoxins in a matrix of hemp (Cannabis sativa L.) seed, and quantification of deoxynivalenol in hemp grain infected with Fusarium graminearum. Journal of the Science of Food and Agriculture. https://doi.org/10.1002/jsf2.70073.
DOI: https://doi.org/10.1002/jsf2.70073

Interpretive Summary: The fungus Fusarium graminearum causes ear rot of corn and head blight of cereal grains. Fusarium graminearum has been found to infect hemp as well. On corn and cereal grains, the fungus produces toxins, including deoxynivalenol (also known as DON or vomitoxin), that can cause vomiting and diarrhea in humans and livestock. Little is known about the production of those toxins in hemp, but because of growing interest in feeding hemp seeds to livestock, it is important to know if hemp seeds infected with F. graminearum may contain these toxins. If the toxins are present in the hemp seeds, a method will be needed to measure the amounts and determine if the amounts are high enough to be a health issue for livestock. A method was developed to measure DON and two structurally similar toxins in hemp seeds. The toxins were separated from each other with the analytical method used. DON could be completely separated from compounds in seed, while the other two toxins were not completely separated from minor compounds in seed. Only DON was found in two samples of harvested hemp grain infected with Fusarium head blight. The method developed in this study would be sensitive enough to measure amounts of DON that could be harmful to livestock. However, it would not be suitable for measuring DON in amounts harmful to humans. Further research is needed to determine if amounts of toxins measured in this study are typical of the amounts found in the field. Further research is also needed to determine if the samples with the most obvious symptoms of disease are also the samples with the most toxins.

Technical Abstract: Background: Fusarium graminearum, one of the causal agents of ear rot in maize and head blight of cereal grains, also infects hemp (Cannabis sativa L. containing a maximum of 0.3% tetrahydrocannabinol or THC). This fungal pathogen produces trichothecene mycotoxins when it infects maize and cereal grains, but little is known about its production of mycotoxins in hemp. A method was developed to separate mycotoxins deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON), and 3-acetyldeoxynivalenol (3-ADON) in commercial hemp seed and grain (harvested tissue), using high-performance liquid chromatography (HPLC) with ultraviolet detection. Results: In HPLC analysis of commercial hemp seed, DON was separated from matrix components. Distortion of the DON peak at higher concentrations led to calibration curves with R-squared values below those of the other mycotoxins analyzed. 15-ADON and 3-ADON slightly overlapped with matrix components. Only DON was detected in two diseased field samples of hemp grain. Conclusions: The difficulty in quantifying one of the hemp grain samples, due to low signal, indicated that the DON concentration of the sample (2.2 µg/g DM) was at the limit of quantitation (LOQ) in the HPLC method developed. Further research is needed to determine if the concentrations of DON in the field samples are representative of mycotoxin concentrations in most samples of symptomatic hemp seed, and to determine if symptom severity is correlated with DON production.