Background: An increased incidence of Barrett’s esophagus in cases with combined acidic and bile reflux is reported. The volunteers serving as controls in those studies were significantly younger than the patients. The aim of this study was to analyze bile reflux patterns of healthy volunteers aged comparably to patients with Barrett’s esophagus. Subjects and Methods: 19 older (8 f, 11 m; median age 51 years) (OV) and 20 younger (10 f, 10 m; median age 25 years) (YV) healthy volunteers without history of gastrointestinal disease or medication underwent simultaneous 24-hour pH and bile monitoring (Bilitec®). All subjects consumed a special diet that did not interfere with bile measurements. Indicators for bile reflux in the stomach and esophagus: bilirubin probe absorption >0.25 in total measuring period (TM), upright position (excluding postprandial periods) (UP) and supine period (SP). Results: There were no differences between older and younger volunteers in pH monitoring. Comparing the periods of bilirubin exposure of the stomach, the median (lower and upper quartile) percentage of time was OV = 5.7% (1.0–15.0%) and YV = 3.5% (0.1–7.8%) respectively for TM (n.s.), OV = 3.1% (0.7–9.4%) and YV = 0.4% (0.0–7.1%) for UP (n.s.), and OV = 2.1% (0.2–16.7%) and YV = 5.9% (0.0–12.2%) respectively for SP (n.s.). In 10/19 older volunteers and in 18/20 younger volunteers, no reflux of bile into the esophagus was measured. However, 4 older volunteers presented bile reflux from 4.6 to 51% of the total measuring period in contrast to the younger group with a maximum of 1.3% of TM. Conclusion: No significant differences in bile reflux into the stomach were distinguishable between younger and older healthy volunteers, but older volunteers more frequently exhibited bile reflux into the esophagus.