Background and study aims: Endoscopic ultrasound (EUS)-guided fine-needle aspiration provides useful diagnostic material in solid and cystic pancreatic lesions. It is logical that EUS-guided pancreatic duct aspiration may be useful in cases of suspected intraductal or duct-obstructing tumors. We evaluated the safety and efficacy of EUS-guided pancreatic duct aspiration. Patients and methods: Twelve patients with dilated pancreatic ducts underwent EUS-guided duct aspiration. Aspirates were submitted for cytology and mucin staining. Patients were followed for up to 13 months. Results: There were no procedure-related complications. Cytology was diagnostic in nine of the 12 patients (six with intraductal papillary mucinous tumor, one with pancreatic clonorchiasis, and two with obstructing solid pancreatic adenocarcinomas). Cytology in the remaining three patients, all with solid obstructing masses, was nondiagnostic. Overall, the diagnostic yield was 75%; however, the yield was 100% in patients without extrinsic obstruction. Conclusion: This preliminary experience suggests that EUS-guided pancreatic duct aspiration is safe and can provide diagnostic material in a significant number of patients with unexplained pancreatic duct dilation.