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Muhammad Subhan Arshad,1 Iltaf Hussain,1 Muhammad Nafees,1 Abdul Majeed,1 Imran Imran,2 Hamid Saeed,3 Furqan K Hashmi,3 Muqarrab Akbar,4 Muhammad Asad Abrar,1 Basit Ramzan,5 Muhammad Omer Chaudhry,6 Muhammad Islam,3 Areeba Usman,7 Naveed Nisar,8 Muhammad Fawad Rasool1 1Department of Pharmacy Practice, Faculty of Pharmacy, Bahauddin Zakariya University, Multan 60800, Pakistan; 2Department of Pharmacology, Faculty of Pharmacy, Bahauddin Zakariya University, Multan 60800, Pakistan; 3University College of Pharmacy, University of the Punjab, Allama Iqbal Campus, Lahore 54000, Pakistan; 4Department of Political Science, Bahauddin Zakariya University, Multan 60800, Pakistan; 5Al Shifa Pharmacy, Multan, Pakistan; 6School of Economics, Bahauddin Zakariya University, Multan 60800, Pakistan; 7Nishter Medical Hospital, Multan, Pakistan; 8Department of Pharmaceutics, Faculty of Pharmacy, Bahauddin Zakariya University, Multan 60800, PakistanCorrespondence: Muhammad Fawad RasoolDepartment of Pharmacy Practice, Faculty of Pharmacy, Bahauddin Zakariya University, Multan, PakistanTel +92 3008639046Email fawadrasool@bzu.edu.pkPurpose: The COVID-19 (coronavirus disease-2019) has been associated with psychological distress during its rapid rise period in Pakistan. The present study aimed to assess the mental health of healthcare workers (HCWs) in the three metropolitan cities of Pakistan.Methods: A cross-sectional, web-based study was conducted in 276 HCWs from April 10, 2020, to June 5, 2020. Depression, anxiety, and stress scale (DASS-21) were used for the mental health assessment of the HCWs. Multivariable logistic regression analysis (MLRA) was performed to measure the association between the demographics and the occurrence of depression, anxiety, and stress (DAS).Results: The frequency of DAS in the HCWs was 10.1%, 25.4%, and 7.3%, respectively. The MLRA showed that the depression in HCWs was significantly associated with the profession (P< 0.001). The anxiety in HCWs was significantly associated with their age (P=0.005), profession (P< 0.05), and residence (P< 0.05). The stress in HCWs was significantly associated with their age (P< 0.05).Limitation: This study was conducted in the early phase of the COVID-19 pandemic, when the number of COVID-19 cases was on the rise in Pakistan and it only represents a definite period (April to June 2020).Conclusion: The symptoms of DAS are present in the HCWs of Pakistan and to manage the psychological health of HCWs, there is a need for the initiation of psychological well-being programs.Keywords: depression, anxiety, stress, pandemic, DASS-21