Lipase activity was measured in supernatant homogenates from various anatomic regions in the upper part of the human digestive tract of two organ donors. It is shown unambiguously that lipase activity occurs only in the fundic mucosa of the stomach, whereas no significant activity takes place in the antral, pharyngeal, or lingual areas, including the circumvallate papillae. In adults, the potential activity of human gastric lipase, as measured using tributyrin as substrate, amounts to 20% of its pancreatic counterpart. Lipase activity was also determined on human gastric biopsy samples taken during gastrofibroscopy tests on healthy adults. These results confirmed the finding that a lipolytic activity of gastric origin occurs uniformly and only in the fundic mucosa. Triacylglycerol hydrolysis is associated with a genuine gastric lipase activity that is clearly distinct from the classical esterase observed using p-nitrophenyl acetate as substrate. Lipase activity decreases significantly with age: it ranges on average from 4700 U/g of fresh mucosa in subjects aged up to 50 yr to 700 U/g of fresh mucosa in persons over 60 yr of age.