Background <p>Health literacy (HL) is the capacity to get, understand, assess, and use health information to prevent disease and improve health in day-to-day living. The study aimed to evaluate the validity and reliability of the Bangla version of the short form of the European Health Literacy Survey Questionnaire (HLS-EU-Q16) for university students.</p> Methods <p>This cross-sectional study was conducted from June 2023 to August 2023. A sample of 948 respondents was selected from three universities in Bangladesh (Jahangirnagar University, Dhaka University, and Ahshanullah University of Science and Technology) using a convenience sampling technique. A structured questionnaire was used to collect data from the respondents. IBM SPSS statistical software, version 25, and AMOS (Analysis of Moment Structure) were used to analyze the data. Scale’s validity and reliability were examined using Cronbach’s alpha coefficient and confirmatory factor analysis, respectively. The chi-square test was used to compare different variables. The level of significance is 0.05.</p> Results <p>The Kaiser-Meyer-Olkin coefficient was 0.826, and the Bartlett test of sphericity was statistically significant (χ² = 3064.93, <i>df</i> = 120, <i>p</i> &lt; 0.001). Confirmatory factor analysis revealed that the three-factor model provided a good fit to the data, yielding better goodness-of-fit outcomes for the Bangla HLS-Q16 (GFI = 0.965, CFI = 0.926, TLI = 0.906, and RMSEA = 0.044) compared to a one-factor model. For the whole scale, Cronbach’s Alpha value was 0.808 within the acceptable limit. All domains had satisfactory internal consistency (Health Care = 0.71, Disease Prevention = 0.70, and Health Promotion = 0.70). In addition, the item-total correlation coefficient varies from <i>r</i> = 0.313 to <i>r</i> = 0.480. All correlations were significant, indicating that each item contributes to the scale score.</p> Conclusion <p>The HLS-Q16 Bangla version is a reliable and valid instrument for evaluating health literacy.</p>