Allulose is a rare sugar that provides <10% of the energy but 70% of the sweetness of sucrose. Allulose has been shown to attenuate glycemic responses to carbohydrate-containing foods in vivo. This study aimed to determine the optimal allulose dose for minimizing in vitro glucose release from rice compared to a rice control and fructose. A triphasic static in vitro digestion method was used to evaluate the in vitro digestion of a rice control compared to the co-digestion of rice with allulose (10 g, 20 g, and 40 g) and fructose (40 g). In vitro glucose release was affected by treatment (p < 0.001), time (p < 0.001), and treatment-by-time interaction (p = 0.002). Allulose (40 g) resulted in a reduction in in vitro glucose release from rice alone and rice digested with allulose (10 g), allulose (20 g), and fructose. The incremental area under the curve (iAUC) for in vitro glucose release was lower after allulose (40 g) (p = 0.005) compared to rice control and allulose (10 g) but did not differ from allulose (20 g) or fructose. This study demonstrates that allulose reduces glucose release from carbohydrates, particularly at higher doses, underscoring its potential as a food ingredient with functional benefits.