The DSM-5 Self-Rated Level 1 Cross-Cutting Symptom Measure was developed to aid in clinical decision-making for clients seeking psychiatric services and to facilitate empirical investigation of the dimensional nature of mental health issues. Preliminary evidence supports its utility with clinical samples. However, the brief, yet comprehensive structure of the DSM-5 Level 1 measure may benefit a high-risk population that is less likely to seek treatment. College students have high rates of hazardous substance use and co-occurring mental health symptoms, yet rarely seek treatment. Therefore, the current study evaluated the psychometric properties (i.e., construct and criterion-related validity) of the DSM-5 Level 1 measure with a large, diverse sample of non-treatment-seeking college/university students. Data from 7,217 college students recruited from 10 universities in 10 different states across the United States evidenced psychometric validation of the DSM-5 Level 1 measure. Specifically, we found acceptable internal consistency across multi-item DSM-5 domains and moderate to strong correlations among domains (internal validity). Further, several DSM-5 domains were positively associated with longer, validated measures of the same mental health construct and had similar strengths of associations with substance use outcomes compared to longer measures of the same construct (convergent validity). Finally, all DSM-5 domains were negatively associated with self-esteem and positively associated with other theoretically relevant constructs, such as posttraumatic stress (criterion-related validity). Taken together, the DSM-5 Level 1 measure appears to be a viable tool for evaluating psychopathology in college students. Several opportunities for clinical application and empirical investigation of the DSM-5 Level 1 measure are discussed.