In the present study, we evaluated whether a low glycemic index (GI) breakfast with lactic acid bread had an effect on glucose tolerance and insulinemia at a subsequent high GI lunch meal. A barley bread containing lactic acid and a reference barley bread were consumed in the morning after an overnight fast in random order by 10 healthy men and women. Four hours after the breakfasts, the subjects ate a standardized high GI lunch, and the blood glucose and insulin responses were measured for the next 3 h. Significant lowerings of the incremental glycemic area (-23%, P = 0.033) and of the glucose response at 95 min were found after the lunch meal when the barley bread with lactic acid was given as a breakfast. At 45 min after the lunch meal, the insulin level was significantly lower (-21%, P = 0.045) after the lactic acid bread breakfast, compared with the barley bread breakfast without lactic acid. We concluded that barley bread containing lactic acid eaten at breakfast has the potential to improve second-meal glucose tolerance at a high GI lunch meal 4 h later.