Background. Hand eczema is common in healthcare workers, owing to intensive exposure to wet work and skin irritants. Targeted interventions and vocational guidance based on documented exposures and risk factors are needed. Objectives. The aims of the study were to investigate the relationship between exposures (domestic and at work) and prevalence and severity of hand eczema. Methods. Self-administered questionnaires were sent to 3181 healthcare workers in Denmark. Results. Two thousand two hundred and sixty-nine (71%) workers responded to the questionnaire. Frequent hand washing was significantly related to the presence of hand eczema. Having children < 4 years old in the household was also related to the presence of hand eczema. A lower prevalence of hand eczema was found among those using moisturizers at work, and a higher prevalence was found among those using moisturizers at home. Conclusions. Although healthcare workers are recommended to use disinfectants when the hands are not visibly dirty, hand washing is still significantly related to hand eczema. Frequent hand washing may be a question of behavioural habits, and a focus for future guidance should be on changing hand washing habits. Attention should also be paid to healthcare workers with small children at home. The preventive effect of moisturizers used during working hours should be tested in future follow-up studies.