University students are a risk group for several mental health problems due to their psychological, social, and academic demands. The aim of this study was to determine the prevalence of symptoms associated with depression, anxiety, and stress among university students from northern Chile and their association with specific socio-demographic characteristics. The study was carried out with 696 participants selected through stratified probabilistic sampling. The depression, anxiety, and stress symptoms were assessed with the DASS-21 scale. The scale's reliability was evaluated with Cronbach's α, and for data analysis, we performed summary measures, frequencies, ANOVA, and Scheffé's test in SPSS 26 ®. Additionally, Cohen's d was calculated for effect size. The results show prevalence of depression was 42.3%, anxiety was 53.5%, and stress was 49.6%. Depression was statistically higher in female than male students with low socioeconomic income and with another sexual orientation from heterosexual. Anxiety was higher in females and in students with low socioeconomic incomes, and, in the case of stress, it was most frequent in women. We found a high prevalence of depression, anxiety, and stress, which highlights the need to develop educational strategies, identify high-risk students, and initiate mental health interventions opportunely.