The beef industry in Poland heavily relies on the Polish Holstein-Friesian (PHF) breed, known for its primary use in dairy production, but which also contributes significantly to the beef supply. In contrast, the Limousine (LM), Hereford (HH), and Charolaise (CH) breeds have gained popularity due to their ideal specialized characteristics for beef production. As PHF continues to dominate the beef market, a thorough comparison of its beef quality and nutritional attributes with the three most popular beef breeds in Poland is essential. This study aims to address this knowledge gap by conducting a rigorous comparison. The experiment was carried out on the beef from 67 bulls kept in a free-stall system with standardized feeding. The highest total antioxidant status (TAS) was found in CH and was 147.5% higher than that in PHF. Also, compared with PHF, a large difference of 70% was observed in LM, while in HH it was only 6.25%. For degree of antioxidant potential (DAP), the highest concentration was found in LM, while CH had a slightly lower score than LM. PHF had the lowest scores for each of the analyzed parameters of protein fraction. For anserine, taurine, creatinine, and creatine content, the highest results were found for LM. For carnosine and coenzyme Q10, the highest values were found for CH. Overall, these results highlight the impact of maturity and breed on carcass composition and quality. Late-maturing breeds, such as LM and CH, tend to exhibit leaner carcasses with superior fatty acid profiles and antioxidant properties. This knowledge is valuable for producers, enabling them to make informed decisions regarding breed selection and production strategies to meet specific market demands for beef with the desired composition and quality.