The aim of this study was to explore the emotional impact of Covid-19 on healthcare professionals at the Clinical and University Hospital of Valencia (Spain), and to identify the associated variables. A total of 228 professionals completed an ad hoc online survey in May 2020. The healthcare professionals experienced symptoms of stress (32%), depression (26%), anxiety (14%) and intrusive memories (7.5%). A higher frequency of sadness and anxiety was observed in women and in professionals who had presented Covid-19 symptoms. The professional category with the most frequent emotional symptoms was that of auxiliary nurses, and for stress also nurses and residents. The most affected were the first line units. The psychological variables that were negatively associated with the frequency of all symptomatic manifestations were: self-care, self-esteem, resilience, and use of active coping strategies, together with self-efficacy and social support for stress and depression. Despite the limitations of the study, the results may contribute to guide preventive programs for health professionals in future health crises.