Acerola fruit is one of the richest natural sources of ascorbic acid. As a consequence, acerola fruit and its products are in demand worldwide for the production of health supplements and for the development of functional products. Acerola phenotypes (103) were screened in Western Paraná State, in the Southern region of Brazil, and evaluated to obtain information on fruit quality characteristics with the aim of using them in future breeding programs. Principal Component and Hierarchical Cluster analysis were performed on all datasets to explore the variability among samples and to identify the main clusters. A great variability among phenotypes was observed, with potential for use in breeding programs. Seven phenotypes were selected as candidates in the next breeding program, characterized by high vitamin C content and yield, or higher values of fruit size and color parameters. Four belong to cluster 1 and three to cluster 2. Specifically, two phenotypes, belonging to cluster 2, showed the best performance in terms of vitamin C (2150 mg 100 g−1 pulp and 2625 mg 100 g−1 pulp respectively) and pulp yield (74.8% and 82.3% respectively), and one phenotype, belonging to cluster 1, for high pulp yield, fruit size and vitamin C content (80.3% 6.43 g and 2490 mg 100 g−1 pulp).