Starch is the main sugar source present in staple foods. Understanding starch hydrolysis during digestion and the resulting glucose release can be important to strategically modulate starch digestion and glucose absorption. In vitro digestion methodologies are fundamental to evaluate starch hydrolysis length and rate, but the lack of uniformity between protocols prevent the comparison of results. In this context, three different Carolino rice varieties (i.e., Carolino white-Cw, Carolino brown-Cb and Carolino Ariete brown-CAb) were submitted to the INFOGEST harmonized in vitro digestion protocol for the evaluation of starch hydrolysis and subsequent glycemic index (GI) determination, and starch granules morphological study. Samples of Carolino rice presented total starch percentages between 64.52 (for Cb) to 71.52% (for Cw) with low amylose content (16.19-19.95%, varying in the following order Cb \ Cab & Cw). During digestion, between 39.43 (for CAb) to 44.48% (for Cb) of starch was hydrolyzed, classifying samples as medium GI foods (61.73-69.17). Starch hydrolysis was accompanied by a decrease of starch granules dimensions. For all samples, area decrease was higher than 59%, perimeter decrease was higher than 37%, feret diameter decrease was higher than 39% and minimum feret diameter decrease was higher than 32%. This work provides new insights to describe, both qualitatively and quantitatively, the fate of rice during digestion, and allowed establishing a comparative basis for the development of rice-based recipes with a lower GI.