Relations between working conditions and mental health status of female hospital workers were studied in a sample of 1505 women: 43% were nurses, 32% auxiliaries, and 7% ancillary staff; 13% were other qualified health care staff, mainly head nurses; 5% had occupations other than direct health care; 63% worked on the morning, 20% on the afternoon, and 17% on the night shift. Data were collected at the annual routine medical visit by the occupational health practitioner, using self administered questionnaires and clinical assessments. Five health indicators were considered: a high score to the general health questionnaire (GHQ); fatigue; sleep impairment; use of antidepressants, sleeping pills, or sedatives; and diagnosis of psychiatric morbidity at clinical assessment. Four indices of stress at work were defined: job stress, mental load, insufficiency in internal training and discussion, and strain caused by schedule. The analysis was conducted by multiple logistic regression, controlling for type of occupation, shift, number of years of work in hospital, daily travel time to work, age, marital status, number of children, and wish to move house. Sleep impairment was mostly linked to shift and strain due to schedule. For all other indicators of mental health impairment and especially high GHQ scores, the adjusted odds ratios increased significantly with the levels of job stress, mental load, and strain due to schedule. This evidence of association between work involving an excessive cumulation of stress factors and mental wellbeing should be considered in interventions aimed at improving the working conditions of hospital workers.