The relative glycemic impact (RGI), the weight of glucose that would induce a glycemic response equivalent to that induced by a given amount of food, is preferably expressed for reference amounts of foods customarily consumed per eating occasion. But because customarily consumed portions of different foods deliver different glycemic carbohydrate doses, methods for determining their RGI need to allow for homeostatic responses to different glycemic carbohydrate loadings. We tested the accuracy of an in vitro method for measuring the RGI of customarily consumed portions that allows for homeostasis, using 24 foods. Glucose equivalents released during simulated gastrointestinal digestion were adjusted by the glycemic potency of contributing sugars to obtain cumulative glycemic glucose equivalents (GGE) and multiplied by food portion weight. Corresponding dose-dependent blood glucose clearance was calculated and subtracted from GGE, giving net GGE compared with time curves reminiscent of blood glucose response curves. RGI values (GGE content) for the food portions were obtained by comparing incremental areas under the curves for foods with that for a white bread reference of known GGE content. The correlation between in vivo values calculated from glycemic index values for the same foods and in vitro values was: in vivo GGE = 1.0 in vitro GGE - 0.5; R2 = 0.90. Bland-Altman methods comparison analysis showed close agreement: in vivo GGE = -0.055 in vitro GGE + 1.16; R2 = 0.027. The results suggest that a modified available carbohydrate determination can economically provide valid RGI values for consumer and industry use.