Early diagnosis of health conditions such as depression, anxiety, and stress, which have been recognized as global health issues, is essential for providing psychological support to people who experience negative emotions. Therefore, it is important to identify a reliable method for diagnosing depression, anxiety, and stress. To this end, this study investigated the factor structure and evaluated the psychometric properties of the Depression Anxiety Stress Scale-21 (DASS-21) in Jordan. A university sample of 336 Jordanians completed the Arabic version of the DASS-21 and several psychopathology measures. Cross-sectional study and confirmatory factor analysis (CFA) were applied to achieve the study's aims. CFA favored the bifactor model compared to the other four models. In contrast, the Beck Anxiety Inventory (BAI) and the Beck Depression Inventory-II (BDI-II) strongly correlated with the DASS-21, indicating that the DASS-21 has adequate convergent and divergent validity. Subsequently, the scale's discriminant validity was tested using the heterotrait-monotrait (HTMT) correlation ratio, which was found to be relatively insufficient. While DASS-21 showed strong reliability, Cronbach's α and McDonald's omega values ranged between 0.83 and 0.93. The bifactor model best fits the Jordanian sample data. The DASS-21 has optimum psychometric properties; therefore, it can be used to assess the general distress experienced by the Jordanian community during research investigations and in non-clinical settings.