Strawberry is a very popular fruit, appreciated for its unique flavor and many beneficial traits such as antioxidants and useful amino acids, which strongly contribute to the overall quality of the product. Indeed, the quality of fresh fruit is a fundamental aspect for consumers, and it is crucial for the success of breeding activities as well as for enhancing the competitiveness and profitability of the fruit industry. Nowadays, the entire supply chain requires simple and fast systems for quality evaluation. In this context, the pomological and chemical traits (i.e., soluble solids, firmness, titratable acidity, dry matter) as well as nutritional ones such as total phenols, total anthocyanins and antioxidant potential were evaluated and compared for seven strawberry cultivars and three harvest times. The prediction of the qualitative traits was carried out using color space coordinates (L*, a* and b*) and two statistical techniques, i.e., the multiple linear regression models (MLR) and artificial neural networks (ANNs). Unsatisfactory prediction performances were obtained for all parameters when MLR was applied. On the contrary, the good prediction of the internal quality attributes, using ANN, was observed, especially for both antioxidant activity and the total monomeric anthocyanin (R² = 0.906, and R² = 0.943, respectively). This study highlighted that color coordinates coupled with ANN can be successfully used to evaluate the quality of strawberry.