It has been previously demonstrated that glucose stimulation of islets of Langerhans causes an accumulation of unesterified arachidonic acid that correlates with insulin secretion. In addition, it is well established that glucose metabolism is essential for insulin secretion. We show that non-metabolizable analogs of glucose which do not stimulate insulin secretion fail to cause significant accumulation of unesterified arachidonic acid. In addition, mannoheptulose, an inhibitor of glucose metabolism, completely blocks the glucose-induced increase in arachidonic acid accumulation. Among the nutrient secretagogues tested, only alpha-ketoisocaproic acid causes a significant increase in unesterified arachidonic acid accumulation. Mannose, fructose, and glyceraldehyde, in particular, failed to elicit a significant increase in unesterified arachidonic acid accumulation. Our data, taken together with previous studies, suggests that glucose must be metabolized to induce accumulation of unesterified arachidonic acid in pancreatic islets.