This epidemiological cohort study, based on Finnish public sector data, investigated the associations between objective working hour characteristics and work-life conflict in day and shift work. The comprehensive data of hospital workers (n = 8 931, 92% women, average age 45 years), consisted of survey responses from 2012, linked with the payroll data of working hour characteristics from 91 days preceding the survey. Logistic regression analysis was used to investigate the associations between working hour characteristics and experiencing work-life conflict often/very often. The analyses were adjusted for age (50 years), sex, level of education, marital status, number of small (0-6 years) and school-aged (7-18 years) children, and the overall stressfulness of the life situation. We also conducted stratified analyses of age and sex on the basis of significant interactions. Difficulties in combining work and life were more often associated with shift work without night shifts and shift work with night shifts than with day work (41% and 34 versus 27%; OR for shift work with night shifts 1.78, 95% CI 1.59-2.00, OR for shift work without night shifts 1.42, 95% CI 1.26-1.60). A high proportion (> 25%) of long (> 40h, (OR 1.26, 95% 1.14-1.39) and very long (> 48h, OR 1.31, 95% CI 1.15-1.49) weekly working hours were associated with work-life conflict, and in the stratified analysis, the latter was also true among women (OR 1.54, 95% CI 1.25-1.89). Of the unsocial working hour characteristics, a relatively large amount (> 10% of all shifts) of evening (OR 1.56, 95% CI 1.41-1.72) and night shifts (OR 1.46, 95%CI 1.32-1.61), a high proportion (> 25% of all shifts) of quick returns ( 25% of all days off) was associated with work-life conflict among men (OR 1.90, 95% CI 1.11-3.25), but not in the whole sample. When the two types of shift work were analyzed separately, shift work without night shifts and very long work weeks had higher odds (OR 1.47, 95% CI 1.20-1.80) of work-life conflict than shift work with night shifts. Conversely, weekend work and evening shifts had higher odds of work-life conflict among shift workers with night shifts (OR 1.74, 95% 1.55-1.96; (OR 1.57, 95% CI 1.40-1.77) than among those without night shifts. To conclude, this study shows that shift workers with and without night shifts more often have difficulties combining work and life than day workers. Several unsocial working hour characteristics, including long work weeks, evening and night shifts, weekend work, and quick returns, are associated with work-life conflict.