This study aimed to perform the cross-cultural adaptation and validation of the eHealth Literacy Scale (eHEALS) for Brazilian Portuguese. The cross-cultural adaptation was conducted through conceptual equivalence, verbatim translation, semantic, item and operational equivalence, and back-translation. Subsequently, 521 undergraduate students answered the adapted version of the eHEALS, the General Perceived Self-Efficacy Scale (GSES), the Oral Health Impact Profile-14 (OHIP-14) and sociodemographic and health-related questions. Statistical analysis was performed to determine the dimensionality, internal consistency, stability and concurrent/convergent/discriminant/predictive validities of the eHEALS (p < 0.05). The adequacy of sample size was confirmed by a non-identity correlation matrix (Bartlett’s test of sphericity, p < 0.001), without influence of multicollinearity (determinant = 0.026). The confirmatory factor analysis identified four factors in agreement to the constructs namely as online searching self-efficacy, awareness of available sources, information usage ability, and critical analysis of useful information. The eHEALS displayed an excellent internal consistency (Cronbach’s alpha = 0.88), a good stability (ICC = 0.71, 95% CI: 0.49–0.84), and it was significantly correlated with GSES (concurrent validity). Higher eHEALS scores were identified among (i) white and (ii) younger students, (iii) from health sciences courses, (iv) with more years of graduation, (v) who their fathers were working in specialized functions (discriminant validity), and (vi) among those who were frequently interested in digital health information (convergent validity). The students with higher eHealth literacy levels were more likely to seek health information in the last 24 h (predictive validity). In conclusion, the eHEALS demonstrated adequate psychometric properties to be applied for the Brazilian population.