Background and Objective: Considering relationship between chronic ankle instability and postural instability, and the importance of the understanding of athletes with chronic instability for rehabilitation clinics, this study was to evaluate functional tests as distinctive tools in individuals with and without chronic ankle instability. Subjects and Methods: This case-control study was conduct on 40 athletes (20 athletes with chronic ankle injury and 20 healthy athletes). In order to distinguish between athletes with chronic ankle instability and healthy athletes, they underwent five functional tests including; Latin-Latin test, side-to-face tests, Lilli square test, zigzag jump test for distance, single jump test for distance. For statistical analysis, test-retest, correlation coefficient and independent t-test were used at the significant level of P <0. 05. Results: Athletes with chronic ankle instability as compared to healthy athletes showed weaker functions. There was a significant difference between participants with chronic ankle instability and healthy participants in performing functional tests (P<0. 05). Significant correlation was found between ankle instability and eight Latin (ICC = 0. 67, p <0. 001), bilateral mutations (ICC = 0. 75, P<0. 001), square jump (ICC=0. 51, P <0. 001), zigzag jump (ICC = 0. 82, P <0. 001). Conclusion: It seems that functional tests (Latin eight test, side leak test, Lilli square test, zigzag jump test for distance, and single jump test for distance) are good tools to identify people with chronic ankle instability from healthy individuals.