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Background Patients with Substance use disorder have distinct personality traits, they were high score in novelty seeking (NS) and sensation seeking and lower in Self-directedness and higher in Self-transcendence, so we aim to investigate the relationships of temperament and characteristics with related some variables such as substance of choice. Design and setting A case–control study enrolling 70 Substance use disorder patients and 70 controls was conducted at Mashhad University of medical sciences. Methods Using a case–control design, a group of 70 Substance use disorder patients and 70 controls was conducted at Mashhad university of medical sciences. All participation completed the 240 questions of Temperament and Character Inventory-Revised (TCI-R). Multivariate analysis of covariance (MANCOVA) was employed to compare the relationship between temperament and character traits and patterns of substance use. Results The scores of reward dependence, persistence, self-directedness, cooperativeness, and self-transcendence were significantly lower in the case group compared to healthy individuals (P P P = 0.637). Conclusions Higher NS in patients with substance use disorder is common and different traits, and temperaments would choose different substance combinations.