According to literature, the incidence of three-canal maxillary first premolar is between 0.5% and 7.5%. The two types of three-canal maxillary premolars are two-rooted and three-rooted three-canal maxillary premolars. Most case reports in Chinese literature focus on three-rooted three-canal maxillary premolars. To our knowledge, no domestic case report on two-rooted three-canal maxillary premolar has been published. In this study, we describe the root canal treatment of a two-rooted three-canal maxillary premolar.