Fusarium head blight occurs worldwide in barley production, and the Fusarium mycotoxins, particularly deoxynivalenol (DON), have become a major concern for barley products, such as beer. This study investigated the content of bound DON in barley samples that were naturally infected with Fusarium head blight. Free DON was determined by gas chromatography-electron capture detection after standard acetonitrile:water extraction, while total DON was determined using treatment with trifluoroacetic acid. Bound DON is the difference between the total and free values. Bound DON was detected in approximately 40% of the samples analyzed, and represented 6 to 21% of free DON. A preliminary study indicated that both free and bound DON decrease significantly during the steeping phase of malting. These results suggest that bound DON, like the free form, may be water-soluble.