the aim of the present study was to validate and implement a nutrition questionnaire to measure nutrition knowledge of university students, and athletes. Male, and female students (N = 476) voluntarily participated in the survey in 2019. Non-health related (n = 156), nutrition students (n = 163), and varsity athletes (n = 157) responded to the questionnaire, twice with 15 days' interval between. in this study, new "general and Sport Nutrition Knowledge" (gSNK) and, two other "Short general Nutrition Knowledge" (SgNK) and, "Short Sport Nutrition Knowledge" (SSNK) questionnaires were used to determine the validation of the survey. one-way ANovA, t Test, Pearson correlation coefficient, and cronbach's α statistics were used to evaluate validity and reliability (p < 0.05). the internal consistency, test-retest reliability, concurrent validity with two similar tools, and construct validity among the groups of students for nutrition knowledge were employed throughout the data analysis. Nutrition students outperformed in all nutrition knowledge sections. respectively, athletes were the second, and non-nutrition related students were the third in sport nutrition (p < 0.05). With regard to general and total nutrition knowledge scores, female students performed significantly better than males (p < 0.01). Modified questionnaire was found to be valid, reliable, and suitable tool for Eurasian university students, and athletes. results also illustrated that the intermittently nutrition educations are required for athletes, as it is recommended in related literature.