OBJECTIVE: This study aimed to use cone-beam computed tomography (CBCT) to evaluate the root anatomy and canal morphology of maxillary premolars in a Chinese population and determine their degree of bilateral symmetry. DESIGN: 774 CBCT images were retrospectively analyzed, representing 1387 maxillary first premolars and 1403 second premolars. The number of roots and canals were recorded. The morphology of root canal systems was determined according to Vertucci's classification. The symmetry of root and canal anatomies between maxillary contralateral premolars was further evaluated. RESULTS: The most common anatomy of maxillary first and second premolars was one-rooted with two canals (58.0%) and one-rooted with one canal (50.3%), respectively. The typical canal morphology was type IV (42.7%) in maxillary first premolars and type I (50.3%) in maxillary second premolars. One-rooted maxillary premolars exhibited a higher variability in the canal morphology, compared to two-rooted or three-rooted teeth. Maxillary second premolars exhibited greater anatomic symmetry than first premolars. The root and canal numbers showed bilateral symmetry between 80.2% of maxillary first premolar pairs and 81.8% of second premolar pairs. Bilateral symmetry in both number and morphology of roots and canals was observed for 72.3% of maxillary first premolar pairs and 73.2% of second premolar pairs. CONCLUSION: The root anatomy and canal morphology of maxillary premolars in a Chinese population were quite diversified. Maxillary contralateral premolars demonstrated a high degree of symmetry in root and canal anatomies, which enables practitioners to better determine the nature of the root canal system during treatment of opposite homonymous teeth.