The effects of infection with Colletotrichum nymphaeae , the causative agent of strawberry black spot, were studied on two strawberry cultivars: susceptible 'Elsanta' and tolerant 'Honeoye' cultivars. Four treatments were established: (1) artificial inoculation; (2) spray application of pyraclostrobin + boscalid (Signum); (3) foliar spraying with calcium (Stopit); and (4) control (spraying with water). Primary metabolites (sugars and organic acids) and secondary metabolites (phenolic compounds) were determined in strawberry fruit with the use of HPLC-MS(n). Infected fruit accumulated large amounts of total sugars and low levels of organic acids. The sugar/acid ratio was higher in the infected and in Ca-treated strawberries. The contents of ellagic acid derivatives, flavonols, oligomeric procyanidins, flavan-3-ols, and total phenolics were highest in inoculated strawberry fruit. Results indicated that fungicide and calcium sprayings did not alter polyphenolic levels in plant tissue.