The purpose of this study was to determine and compare thigh muscle volumes (MVs), and sprint mechanical properties and performance between male and female national-level sprinters. We also studied possible relationships between thigh MVs and sprint performance. Nine male and eight female national-level sprinters participated in the study. T1-weighted magnetic resonance images of the thighs were obtained to determine MVs of quadriceps, hamstrings and adductors. Sprint performance was measured as the time to cover 40 and 80 m. Instantaneous sprint velocity was measured by radar to obtain theoretical maximum force (F0), theoretical maximum velocity (V0) and maximum power (Pmax). When MVs were normalized by height-mass, males showed larger hamstrings (13.5%, ES = 1.26, P < 0.05) compared with females, while quadriceps and adductors showed no statistically significant differences. Males were extremely faster than females in 40 m (14%, ES = 6.68, P < 0.001) and in 80 m (15%, ES = 5.01, P < 0.001. Males also showed increased sprint mechanical properties, with larger F0 (19%, ES = 1.98, P < 0.01), much larger Pmax (46%, ES = 3.76, P < 0.001), and extremely larger V0 (23%, ES = 6.97, P < 0.001). With the pooled data, hamstring and adductor MVs correlated strongly (r = -0.685, P < 0.01) and moderately (r = -0.530, P < 0.05), respectively, with sprint performance; while quadriceps showed no association. The sex-stratified analysis showed weaker associations compared with pooled data, most likely due to small sample size. In conclusion, males were faster than females and showed larger MVs, especially in hamstrings. Moreover, regarding the thigh muscles, hamstrings MV seems the most related with sprint performance as previously proposed.