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Objectives: There are no validated instruments to measure education-related stress in Brazilian university students. Thus, we aimed to translate and test the internal reliability, convergent/discriminant validity, and measurement equivalence of the Higher Education Stress Inventory (HESI)., Methods: The translation protocol was carried out by two independent translators. The instrument was culturally adapted after a pilot version was administered to 36 university students. The final version (HESI-Br) was administered to 1,021 university students (mean age = 28.3, standard deviation [SD] = 9.6, 76.7% female) via an online survey that lasted from September 1 to October 15, 2020. The factor structure was estimated using exploratory factor analysis (EFA) on the first half of the dataset. We tested the best EFA-derived model with confirmatory factor analysis (CFA) on the second half. Convergent/discriminant validity was tested using the Depression, Anxiety and Stress Scale (DASS-21). Sex, age groups, period of study, family income and area of study were used to test measurement equivalence., Results: EFA suggested five factors: career dissatisfaction; faculty shortcomings; high workload; financial concerns; and toxic learning environment. CFA supported the five-factor model (15 items), but not a higher order factor, suggesting multidimensionality. All five factors presented acceptable internal reliabilities, with Cronbach's α ≥ 0.72 and McDonald's ω ≥ 0.64. CFA models indicated that the HESI-Br and DASS-21 assess different but correlated underlying latent constructs, supporting discriminant validity. Equivalence was ascertained for all tested groups., Conclusion: The 15-item HESI-Br is a reliable and invariant multidimensional instrument for assessing relevant stressors among university students in Brazil., Competing Interests: No conflicts of interest declared concerning the publication of this article.