Objective: To assess health-related quality of life (QoL) and its associated factors among health workers (HWs) in tertiary care hospitals of Peshawar, Pakistan.Methods: This analytical cross-sectional study was conducted from February to May 2024 in Medicine & Allied and Surgical & Allied departments of three public and six private tertiary care hospitals in Peshawar. Using non-probability convenience sampling, 365 HWs, including specialists, trainee medical officers, house officers, and nurses, were recruited. Health-related QoL was assessed using World Health Organization Quality of Life-BREF (WHOQOL-BREF) questionnaire. Data were analysed using descriptive and inferential statistics, with p<0.05 considered statistically significant.Results: Among 365 health workers, majority (n=211, 57.8%) were male, with overall mean age of 29.22±6.49 years. The WHOQOL-BREF demonstrated good internal consistency (Cronbach's α=0.883). Mean scores for the physical, psychological, social relationships, and environmental domains were 50.96±14.99, 58.31±16.40, 61.97±23.28, and 54.27±17.83, respectively. Male gender, married status, and employment in Medicine and Allied departments were significantly associated with higher social relationship scores. Nurses had higher physical health scores than other professional groups (p=0.010). Morning shifts, working <8 hours/day, and private-sector employment were associated with better QoL in selected domains. Age was weakly associated with social relationships (r=0.110, p=0.036), while commute duration had no significant impact on any domain.Conclusion: Health-related QoL among HWs varied across WHOQOL-BREF domains and was significantly associated with demographic and occupational factors, particularly gender, professional designation, department, work shift, shift duration, and hospital sector. Workplace interventions targeting modifiable occupational factors may improve well-being and QoL of HWs.