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Objective: The objective of this study was to characterize the structure, physicochemical and toxicity properties of Dioscorea Pentaphylla tuber’s starch. Methods: Starch was extracted from Dioscorea Pentaphylla tuber’s and analyzed to determine their properties such as pH, water binding capacity, solubility, swelling power, chemical composition, thermal properties, degree of crystallinity, morphology and toxicity. Results: Results showed that more than half of Dioscorea pentaphylla starch consist of amylose (64.10 ± 1.15%). It has gelatinization temperature started at 76.90 ± 0.08 °C. This C-Type starch has a degree of crystallinity around 32.90 ± 2.59%. The SEM image showed that Dioscorea pentaphylla has oval-shaped particles with sizes ranging from 42 to 55 µm. The toxicity test demonstrated that the starch was safe and can be classified as non-toxic. Conclusion: The physicochemical and toxicology properties of Dioscorea Pentaphylla starch recorded in this study indicate that this starch is applicable as biopolymer source for food and non-food purposes.