North Dakota produces just over half of the U.S. durum wheat (Triticum durum Desf.). However, production is frequently threatened by Fusarium Head Blight (FHB) and associated contamination with the mycotoxin deoxynivalenol (DON), due to limited host resistance to Fusarium graminearum. The episodic nature of infections and variable environmental conditions further complicate disease development and DON accumulation. This study aimed to assess DON levels in 21 durum wheat varieties from variety trials conducted at nine locations across North Dakota — Carrington, Garrison, Hettinger, Langdon, Minot, Mohall, Nesson, Rugby, and Williston — during the 2023 and 2024 growing seasons. Grain samples were milled, and DON concentrations were quantified using Neogen’s Reveal Q⁺ DON test strips, with results read by the AccuScan Gold® Lateral Flow Strip Reader. Data were analyzed using JMP® Student Edition 18.2.2. Results showed a significant Variety × Location × Year interaction (P < 0.0001). While varietal differences in DON levels were not statistically significant (P = 0.1084), location had a strong influence (P = 0.0040), with DON levels notably higher in Rugby and Mohall in 2024, while Langdon remained stable across both years. These findings underscore the challenges posed by inconsistent disease pressure and environmental variability, emphasizing the need for continued statewide monitoring of DON. Moreover, locations like Rugby and Mohall may benefit from more aggressive FHB management strategies, including the use of the FHB Risk Tool and timely fungicide applications, to reduce DON contamination. These insights are valuable for informing durum variety selection and guiding site-specific disease management across North Dakota.