Objective: to analyze the association between socio-occupational characteristics, stress level, and quality of life in hospital Nursing workers. Method: a cross-sectional, quantitative study, with data collected by means of a socio-occupational questionnaire to survey the profile of the workers, the Bianchi Stress Scale to assess stress, and the WHOQOL-Bref to assess quality of life. Results: 180 Nursing workers participated in the research, being 49 nurses and 131 Nursing technicians and assistants, most of them women, predominantly aged up to 35 years old, married, and with up to three years of experience in the sector. In general, the participants presented medium stress level, with a higher level among the nurses, and greater satisfaction with the quality of life level related to the physical (65.6%), psychological (64.7%), and social relationships (67.9%) domains. This study identified significant associations between stress and quality of life in hospital Nursing workers. Conclusion: it was demonstrated that there was an association between stress and quality of life, and the greater the stress, the lower the quality of life in hospital Nursing workers.