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Abstract Background University students majoring in different disciplines are believed to have different personality traits, courses exposure, and future roles, which may further affect their health behaviors and health status. The purpose of this study was to investigate the differences in health-promoting lifestyle (HPL) and its predictors among health-related and non-health-related students. Methods The research participants were university students in the main island of Taiwan, and a two-stage sampling approach was adopted to obtain the samples from November 2020 to March 2021. First, 37 universities were randomly selected based on the ratio of public and private universities in each region of Taiwan. Then, based on the ratio of health-related and non-health-related majors of selected university, 25–30 students were randomly drawn from each university according to the student ID number to complete self-administered questionnaires, which included items for personal factors, perceived health status (PHS), health conception (HC), and health-promoting lifestyle profile (HPLP). A total of 1062 valid questionnaires were recovered, including 458 from health-related students and 604 from non-health-related students. Chi-squared test, independent samples t-test, one-way ANOVA, Pearson product-moment correlation analysis, and multiple regression analysis were performed. Results The results showed that gender (p |