This study evaluated the effects of different pre-slaughter road transportation times on sensory evaluation and instrumental measurements of Nellore cattle meat quality. About 15 farms were classified according to transportation times (five farms per transportation time group) as follows: 1 — less than 2 h (<2), 2 — between 2 and 4 h (>2 and ≤4), and 3 — between 4 and 6 h (>4 and ≤6). The experiment was a completely randomized design with 15 replications and three treatments. Meat analyses were performed on steaks taken from the longissimus dorsi muscle from the 10th rib to the third lumbar vertebra. Road transportation times had no significant effects on pH and mean shear force values. Sarcomere length, myofibrillar fragmentation index, and creatine kinase levels were also not affected by transportation time. Furthermore, road transportation times had no effect on meat tenderness, flavour, odour, and overall acceptability when evaluated by consumers. Thus, a pre-slaughter road transportation time up to 6 h does not affect the sensory evaluation and instrumental measurements of Nellore cattle meat quality when raised using a grazing system.