Introduction: COVID-19 pandemic has affected HCPs in multiple way. It has caused psychological impact in form of anxiety, depression, and insomnia. In this study, we aim to study and compare the stress level, anxiety and depression among HCPs who are posted in special COVID-19 units with the HCPs who are not posted in COVID-19 units.Methods: This cross-sectional study was conducted in June 2020, at various hospitals of Karachi, Pakistan. All health care professionals (HCPs) were invited to participate. A total of 301 HCPs completed this study, who were divided into two groups; those who are posted in COVID-19 ward (Group A) and those who are not (Group B). Psychological Impact was English version of the Depression Anxiety Stress Scale - 21 (DASS-21).Results: In Group A, 70.5% had moderate, severe, or extremely severe depression compared to 49.2% in group B. In Group A, 75.4% had moderate, severe, or extremely severe anxiety compared to 44.7% in group B. In Group A, 80.3% had moderate, severe or extremely severe stress compared to 54.2% in group B. Anxiety, depression and stress were significantly higher in HCPs who were posted in COVID-19 ward compared to those who were not posted in COVID-19 wardConclusion: There was significantly higher anxiety, stress and depression in health care professionals posted in COVID-19 ward. Both the government and health care agencies should take responsibility for protecting the psychological well-being of health care communities all over the world and ensuring a healthy work environment.