Objective: Segmentation of populations may facilitate development of targeted substance abuse prevention programs. The authors aimed to partition a national sample of university students according to profiles based on substance use. Participants: The authors used 2008-2009 data from the National College Health Assessment from the American College Health Association. The sample consisted of 111,245 individuals from 158 institutions. Methods: The sample was partitioned using cluster analysis according to current substance use behaviors. The association of cluster membership with individual and institutional characteristics was examined. Results: Cluster analysis yielded 6 distinct clusters. Three individual factors--gender, year in school, and fraternity/sorority membership--were the most strongly associated with cluster membership. Conclusions: In a large sample of university students, the authors were able to identify 6 distinct patterns of substance abuse. It may be valuable to target specific populations of college-aged substance users based on individual factors. However, comprehensive intervention will require a multifaceted approach. (Contains 5 tables.)