Background. Pancreaticobiliary reflux is a pathologic phenomenon occurring in patients with gallstones. However, the occurrence of pancreaticobiliary reflux has not been studied in patients without gallstones. The objective of this study was to measure the bile levels of amylase and lipase in patients without gallstones submitted to cholecystectomy as part of another surgical procedure, and to compare these values with patients submitted to cholecystectomy for gallstone disease. Patients and Methods. A prospective observational comparative study was designed. A sample of 136 consecutive patients was included. Amylase and lipase levels were measured in bile. Normal serum amylase levels at our institution are 28-100 U/L and for lipase are 13-60 U/L. There are no established normal levels for pancreatic enzymes in bile. However, we considered elevated the bile amylase and lipase levels whenever they were higher than normal plasma levels. Results. One-hundred three patients (76%) had gallstones and 33 (24%) liad healthy gallbladders without gallstones. According to normal plasma levels for amylase and lipase, these enzymes in bile were elevated in 83.5% patients with gallstones, compared to elevated levels of amylase in 6% patients and lipase in 3% patients without gallstones. Conclusions. Pancreaticobiliary reflux is a common phenomenon in patients with gallstones and occurs sporadically in patients without gallstones.