Abstract Background More and more samples are received by the laboratories each day for the simultaneous measurement of serum amylase and lipase in the diagnosis of acute pancreatitis. Objectives This study is an effort to find which of the two analytes, serum amylase or lipase, is a better marker. Methods This is a retrospective study from a tertiary care hospital in which radiologically diagnosed patients of acute pancreatitis, in whom blood samples for the estimation of serum amylase and lipase were collected within 12–72 h after the onset of abdominal pain were taken up for the study. All the relevant data and imaging details were collected. Results We analyzed 100 such cases and in our study we found that 74 out of 100 patients had raised serum amylase and 93 out of 100 had raised serum lipase levels. Conclusion In rural health care centers, in smaller hospitals, or in set ups where resources are limited and in situations where patients cannot afford, serum lipase will be a better choice over serum amylase in the diagnosis of acute pancreatitis.