Location: Forage-animal Production Research
Title: Separation and quantification of mycotoxins in floral tissue of hemp (Cannabis sativa) infected with Fusarium graminearumAuthor
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Kagan, Isabelle |
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SMITH, HENRY - University Of Kentucky |
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SCHENDEL, RACHEL - University Of Kentucky |
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GAUTHIER, NICOLE - University Of Kentucky |
Submitted to: Journal of the Science of Food and Agriculture
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 1/8/2025 Publication Date: 1/27/2025 Citation: Kagan, I., Smith, H.S., Schendel, R.R., Gauthier, N. 2025. Separation and quantification of mycotoxins in floral tissue of hemp (Cannabis sativa) infected with Fusarium graminearum. Journal of the Science of Food and Agriculture. JSFA Reports p. 1–9. https://doi.org/10.1002/jsf2.235. DOI: https://doi.org/10.1002/jsf2.235 Interpretive Summary: Fusarium graminearum is a fungus that causes disease on wheat and that produces toxins when it infects wheat and other grains. This fungus has been found on hemp, and whether it produces the same toxins on hemp as on wheat is uncertain. A method was developed to detect several toxins in hemp floral tissue. Deoxynivalenol (DON, also known as vomitoxin), 15-acetyldeoxynivalenol (15-ADON), and 3-acetyldeoxynivalenol (3-ADON) were separated from cannabinoids by liquid chromatography. DON was completely separated from plant compounds, but 15-ADON and 3-ADON were not completely separated. In tissue infected with F. graminearum, DON or 15-ADON were found, as well as a sample with no DON or 15-ADON. The extraction method recovered about 75% of the 15-ADON present in tissue to which toxins were added, and 86% of the DON and 3-ADON present. The method would be able to detect about 3 µg DON and 15-ADON per gram of tissue, and about 6 µg 3-ADON per gram of tissue. Future work will involve trying to make the method more sensitive. Technical Abstract: Background: The fungal pathogen Fusarium graminearum produces mycotoxins when it infects grains. This pathogen also infects hemp (Cannabis sativa L. containing less than 0.3% tetrahydrocannabinol or THC). The presence of trichothecene mycotoxins like deoxynivalenol (DON) has not been reported in floral tissue of hemp infected with F. graminearum. The object of this study was to develop a method to extract three trichothecene mycotoxins—DON, 3-acetyldeoxynivalenol (3-ADON), and 15-acetyldeoxynivalenol (15-ADON)—from hemp floral tissue, and to quantify the mycotoxins by high-performance liquid chromatography (HPLC) with ultraviolet detection. Results: Standards of nivalenol (NIV), DON-3-glucoside (D3G), DON, 3-ADON, and 15-ADON were separated from each other and from standards of several cannabinoids. Standard curves were linear (R-squared values > 0.99). Recoveries from hemp spiked with 36 µg/g mycotoxins were 86 to 90% for DON, 75 to 77% for 15-ADON, and 86 to 101% for 3-ADON. In extracts of hemp inflorescences infected with different strains of F. graminearum (15-ADON chemotypes), DON was detected in one sample. Other samples contained 15-ADON but no detectable DON, or neither mycotoxin in detectable amounts. Conclusions: Mycotoxins were separated from cannabinoids. DON was also separated from matrix components, and 15-ADON and 3-ADON were partially separated from matrix components. The presence of DON in hemp may depend on fungal strain or stage of disease development, as well as the sensitivity of the method. The current method could quantify DON and 15-ADON at about 3 µg/g in hemp floral tissue, and 3-ADON at about 6 µg/g. |