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Objectives: The aim of this study was to evaluate the predictive value of the first postoperative day (POD1) drain fluid amylase in predicting pancreatic fistula formation following pancreaticoduodenectomy (PD)., Material and Methods: One-hundred and eighty-five prospective patients undergoing PD between April 2014 and April 2018 were studied retrospectively. Cut-off point to predict the development of POPF was determined by median values for drain fluid amylase of 1883 U/L. Patients were classified into two groups according to POD1 drain fluid amylase values: <1883 U/L (Group 1) and ≥1883 U/L (Group 2). Differences between the groups with clinically relevant POPF and without POPF were evaluated., Results: The incidence of POPF was 17.2%. POD1 amylase level was the strongest predictor of POPF, with levels of higher than 1883 U/L demonstrating the best accuracy (87.5%), sensitivity (78.1%), specificity (89.5%), positive predictive value (60.9%), and negative predictive value (95.1%). One-hundred and forty-four patients (77.8%) had a POD1 drain amylase level of less than 1883 U/L, and POPF developed in only seven (3.7%) cases, whereas in patients with POD1 drain amylase level of 1883 U/L or higher (n= 41), the POPF rate was 31.4% [OR: 22.24, 95% CI (7.930-62.396), p<0.001]., Conclusion: The cut-off point of POD1 drain fluid amylase level (1883 U/L) might predict the clinically relevant POPF with adequate sensitivity and specificity rates in patients undergoing pancreatic resection., Competing Interests: Conflict of Interest: The authors have no conflicts of interest to declare., (Copyright © 2024, Turkish Surgical Society.)