Abstract. Endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) is an accurate technique for sampling the pancreas and mediastinum; however, limited data are available for other mass lesions. The aim of this study was to explore the value of EUS-FNA in the differential diagnosis of all mass lesions. Data from patients who underwent EUS-FNA for the diagnosis of mass lesions, including pancreatic, mediastinal, celiac and retroperitoneal lesions were retrospectively analyzed. The accuracy was calculated by comparing the results of FNA with the results of pathological examination or follow-up surveillances in non-operated cases. A total of 150 cases were included. The location of the mass varied from the pancreas (n=62) to the mediastinum (n=29), gastrointestinal tract (n=36), celiac cavity and retroperitoneum (n=23). The sensitivity and Youden ' s index of EUS-FNA in the diagnosis of all lesions were 92.97% and 0.93 respectively. The accuracy of diagnosis of pancreatic, mediastinal, gastrointestinal, celiac and retroperitoneal lesions was 85.48, 89.66, 83.33 and 78.23%, respectively. Masses were categorized into parenchymal organs (n=66), luminal organs (n=36) and enlarged lymph nodes (n=33). Lesions in parenchymal organs were likely to be bigger than those in luminal organs (P=0.03) and enlarged lymph nodes (P=0.01). For solid and cystic masses, which constituted 63.3 and 14.7% of the total masses, no significant difference in diagnostic accuracy was observed (P=0.56); however, lesion sizes were significantly different between these two groups (P=0.04) and the majority of cystic masses were identified in women (P=0.03). Malignant lesions were more common in older (P=0.01) and male (P=0.03) patients. In conclusion, EUS-FNA is an effective tool in the diagnosis of unexplained mass lesions; it influences the management of patients by enabling the appropriate treatment to be identified.