Background: The focus on hand hygiene during the pandemic has been reported to increase the hand eczema (HE) prevalence in health care workers (HCWs); however, detailed prospective data are missing. Objective: To evaluate changes in HE prevalence, exposures, and health-related quality of life among HCWs during the COVID-19 pandemic. Methods: In this prospective cohort study, HCWs employed at the hospitals in Copenhagen responded to a digital questionnaire at the beginning of the pandemic and 11 months thereafter. Results: A total of 795 HCWs responded to both questionnaires (83.4% women). The calculated 1-year HE prevalence decreased from 16.0% at baseline to 13.0% at follow-up. The number of hand washings decreased significantly, whereas the use of alcohol-based hand rubs on wet skin increased significantly. In a logistic regression model, increased use of alcohol-based hand rubs on wet skin was associated with HE at follow-up (odds ratio, 1.78; 95% CI, 1.11-2.87). Health-related quality of life worsened slightly at follow-up, with HE severity and frequent flareups being risk factors for a reduced health-related quality of life. Limitations: Sample size. Conclusion: In contrast to previous studies undertaken during the pandemic, we found a relatively low and stable HE prevalence. Our findings suggest that the interaction between changed exposures and HE is complex and cannot be linked to a single factor.