Background Dietary acrylamide is found in certain foods, such as deep frying, baking and roasting, and is associated with higher inflammatory and oxidative stress parameters. The association between dietary acrylamide and physical performance has not yet been explored. The aim of the study was to investigate the relationship between dietary acrylamide intake and physical performance tests in a large cohort of North American individuals affected by knee osteoarthritis or at high risk for this condition. Methods Dietary acrylamide intake was obtained through a food frequency questionnaire and reported in quartiles and as an increase in deciles. Physical performance was explored using the 20-meter usual pace test, the 400-meter walking distance, and the chair stands time. The association between dietary acrylamide and physical performance tests was explored using linear regression analysis, adjusted for potential confounders. Results 4,436 participants (2,578 women, mean age: 61.3) were enrolled. People in the highest quartile of dietary acrylamide reported significantly longer 20-meter walking (15.53±3.32 vs. 15.15±2.91 s), 400-meter walking (312±54 vs. 305±58 s) and chair stands (11.36±4.08 vs. 10.67±3.50 s) times than their counterparts in Q1. In adjusted linear regression analyses, each increase in one decile in dietary acrylamide was associated with a longer time in walking for 20 meters (beta = 0.032; 95%CI: 0.016–0.048; p = 0.04), 400 meters (beta = 0.048; 95%CI: 0.033–0.063; p = 0.002) and chair stands (beta = 0.016; 95%CI: 0.005–0.037; p = 0.04) times. Conclusion Higher dietary acrylamide intake was significantly associated with poor physical performance, also after accounting for potential confounders, suggesting a role for this food contaminant as a possible risk factor for sarcopenia.