Background: Psychological impact of COVID-19 pandemic on health care workers (HCWs) is a burning issue that needs to be taken care of with utmost importance. The main objective of this study was to assess depression, anxiety and stress among HCWs at a tertiary care hospital dedicated for COVID-19 while fighting with the ongoing second wave of Covid. Methods: A descriptive cross-sectional study was conducted among 245 HCWs at a COVID dedicated hospital in July and August 2021, after IRC approval and with informed written consents. Convenient sampling method was applied. Depression, anxiety and stress scale (DASS) 21 and semi structured proforma were utilized. All the statistical calculations were done by STATA 15.1. Frequency and percentage were calculated as descriptive statistics for baseline characteristics. Results: Depression was observed in 22.86%, anxiety in 50.61% and stress in 15.51%. Stress was highest among nurses (18.18%), followed by clinicians (16.36%). Anxiety was seen highest among nurses (63.64%) followed by pharmacists (60%). Depression was highly prevalent in nurses (26.57%) compared with clinicians (24.45%) and laboratory staff (6.67%). Conclusions: Despite developments in the knowledge, management, exposure and experience with COVID-19 since first wave, depression, anxiety and stress has affected health care workers. Therefore, mental health needs of the health care workers have to be taken care of for maintaining a better effective healthy workforce and service delivery.