Potatoes are generally regarded as high glycemic index (GI) foods. Resistant starch (RS) comprises the starch fraction that is not absorbed in the small intestine, thus controlling the glucose level and improving the intestinal environment. In this study, an analysis of the formation of RS of potato starch samples under different acetic acid-thermal treatment conditions was conducted. Additionally, the relationship between the rates of starch digestion, estimated GI (eGI), and the RS content was evaluated by employing in vitro enzymatic models. Compared with control samples, the RS content in the cold-stored samples after acid-boiling was higher, whereas that of samples after heating at 120 °C with acetic acid was decreased. The eGI was negatively correlated with the RS content in potatoes. Cold store after acid-boiling was effective in increasing the RS content. Furthermore, low eGI values may have resulted from higher levels of RS in potatoes.