Aim: This study aimed to assess the diagnostic accuracy of serum amylase and lipase in acute pancreatitis and perform its cost analysis to check if one alone is sufficient for a more cost-effective diagnostic process.Materials and Methods: This was a retrospective study conducted in a hospital with a 335,000 annual number of emergency department (ED) visits. All patients to whom both amylase and lipase test were performed in 2019 were analyzed. Patients with three or more times the normal range elevation of either the amylase (>300 U/L) or lipase (>195 U/L) were included into the study. Sensitivity of those enzymes in diagnosing acute pancreatitis was calculated. Cost measurement of amylase and lipase was also determined. Results: The number of patients with both enzyme levels measured for any reason at ED was 53,944 in a year. A total of 130 patients who met the inclusion criteria were analyzed. Wherein, 108 had elevated levels of both enzymes. Moreover, 22 of these patients had amylase lower than the three fold of the normal range with a significantly high lipase. No patient had elevated amylase with normal lipase levels. Sensitivity of serum amylase and lipase was 84% and 100%, respectively. If lipase is measured only for all patients whose amylase and lipase were measured, potential saving is calculated as 71,745 TL (10,298 USD) annually.Conclusion: “Lipase only” measurement is recommended in terms of diagnostic accuracy and cost effectiveness in differential diagnosis for acute pancreatitis.