This aim of this study was to evaluate the effect of high-frequency focused ultrasound (HFFU) on quality traits of bovine Triceps brachii. Four treatments (0, 10, 20, and 30 min) of HFFU (2 MHz and 1.5 W/cm²) were applied to bovine T. brachii muscle. Immediately after treatment, evaluations of color, pH, drip loss, water holding capacity, and shear force in meat were undertaken. The application of HFFU slightly decreased (p < 0.05) the redness of meat. In addition, a significant (p < 0.05) decrease in the shear force of meat was observed after the application of HFFU at 30 min. No effect (p > 0.05) was observed on other color parameters, drip loss, and water holding capacity of meat. Overall, HFFU improved beef tenderness without negative impacts on color, pH, drip loss, and water holding capacity of meat. HFFU offers the option of tenderizing specific muscles or anatomical regions of the beef carcass. These findings provide new insights into the potential application of ultrasound in meat processing.