The emerging role of endothelial inflammation in diabetes has stimulated research interest in the effects of nutrition on related indices. In the current study we investigated whether the nutrient composition of dietary formula as reflected in glycemic index (GI) may be predictive of postprandial endothelial inflammation in non-diabetic subjects. A double-blinded, randomized, crossover study was conducted in non-diabetic subjects (n = 8/group). Each subject consumed three types of diabetes-specific dietary formulas (high-fiber formula [FF], high-monounsaturated fatty acid (MUFA) formula [MF] and control formula [CF]) standardized to 50 g of available carbohydrates with a 1-week interval between each. The mean glycemic index (GI) was calculated and 3-hour postprandial responses of insulin, soluble intercellular adhesion molecule-1 (sICAM-1), nitrotyrosine (NT) and free fatty acids (FFA) were measured. The MF showed the lowest mean GI and significantly low area under the curve (AUC) for insulin (P = 0.038), but significantly high AUCs for sICAM-1 (P<0.001) and FFA (P < 0.001) as compared to the CF and FF. The FF showed intermediate mean GI, but significantly low AUC for NT (P<0.001) as compared to the CF and MF. The mean GI was not positively correlated to any of the inflammatory markers evaluated, and in fact negatively correlated to changes in FFA (r = -0.473, P = 0.006). While the MF with the lowest GI showed the highest values in most of the inflammatory markers measured, the FF with intermediate GI had a modest beneficial effect on endothelial inflammation. These results suggest that nutrient composition of dietary formula as reflected in the GI may differently influence acute postprandial inflammation in non-diabetic subjects.