Strawberries are one of the most important food crops grown in Florida, with a harvested area of ≈10,000 acres. The University of Florida strawberry breeding program develops cultivars adapted to this region and its particular weather conditions, with a major aim of increasing overall quality. The objective of this study was to compare the fruit of advanced breeding selections to those of commercial cultivars, for compositional attributes. Seven different strawberry genotypes were compared at harvest and after 7 days at 4 °C across multiple harvest dates during two consecutive years. Compositional attributes were highly influenced by year, harvest date within a year, genotype, and storage. Overall, compared with other genotypes, selection FL 09-127 exhibited consistently higher soluble solids and total sugar (TS) contents at harvest and after cold storage. Higher ascorbic acid (AA) and phenolic contents at harvest were observed in selection FL 07-193. However, its anthocyanin content was among the lowest. In contrast, FL 10-47 exhibited relatively low AA content at harvest but consistently high total anthocyanins (TACs) and total phenolic (TP) contents after storage. Overall, results from this study provide valuable information to the breeding process by identifying new genotypes with improved compositional attributes combined with suitable quality characteristics after cold storage.