The main and interactive effect of biological maturity and relative age upon physical performance in adolescent male soccer players was considered. Consistent with previous research, it was hypothesised that participants of greater maturity or born earlier in the selection year would perform better in terms of physical performance tests. This cross-sectional study consisted of 84 male participants aged between 11.3 and 16.2 years from a professional soccer academy in the English Premier League. Date of birth, height, weight, and parental height were collected. Sprint, change of direction, countermovement jump, and reactive strength index were considered for physical performance. Relative age was based on the birth quarter for the selection year. Maturity status was based upon the percentage of predicted adult height attained. Linear regression models highlighted that maturation was associated with performance on all but one of the physical performance tests, the reactive strength index. In contrast, relative age only served as a significant predictor of performance on the countermovement jump. This study indicated that physical performance (in the tests studied) seems to be related to the biological maturity status of a player but not their relative age. This finding is important because it suggests that early-maturing players perform better in the majority of physical performance tests, and the commonly held belief that relative age effect influences performance may be overstated.