This work aimed to evaluate the use of annealing (ANN) ulluco starch in the preparation of biodegradable films and its impact on the physicochemical properties of the materials. Three film samples (FS1, FS2, and FS3) were prepared at a fixed starch concentration (2.6% w/v) using glycerol as a plasticizer and then compared to a control sample (FSC) prepared with native ulluco starch. The physical, mechanical, and thermal properties of the films were evaluated. The use of ANN starch decreased the solubility (from 21.8% to 19.5%) and the swelling power (from 299% to 153%) of the film samples. In addition, an increase in opacity and relative crystallinity (from 7.54% to 10.5%) were observed. Regarding the thermal properties, all the samples presented high stability to degradation, with degradation temperatures above 200 °C. However, the samples showed deficiencies in their morphology, which affected the barrier properties. The use of ANN starch has some advantages over native starch in preparing films. However, more analysis is needed to improve the barrier properties of the materials. This work reveals the potential of the ANN ulluco starch for biodegradable film preparation. In addition, the use of modified ulluco starch is an alternative to add value to the crop, as well as to replace non-biodegradable materials used in the preparation of packaging.