To explore the dietary intake risk of deoxicallenol (DON) from barley subjected to different storage conditions or processing methods, barley was stored for different periods (180, 270 and 360 days) at different temperature (5, 15, 25 and 35 ℃) and relative humidities (55%, 65%, 75% and 85%). Another part of samples were processed by boiling, steaming or fermentation. DON content in all samples was detected by enzyme-linked immunosorbent assay (ELISA), and the risk of chronic and acute dietary intake of DON was explored by point and probability assessment. The results showed that the estimated daily intake (EDI) of DON from fresh barley was 0.00030 to 0.00130 mg/kg, and the chronic risk index was 0.30 to 1.31, which increased by up to 0.63 after 360 days of storage. The risk of dietary exposure to DON was 32.7%, the national estimated short-term intake (NESTI) was 0.0026 to 0.0055 mg/kg and the hazard quotient (HQ) for acute riskassessment was 0.33 to 0.69, indicating no acute exposure risk. After processing, the risk of chronic exposure decreased from 0.60–2.60 to 0.17–2.34, and the risk of acute exposure decreased by up to 0.64. After long-term storage, the level of exposure to DON from barley was high, causing a high health risk, which was effectively reduced after processing, and the exposure risk for children was more serious than that in adults, to which much attention should be paid.