Physical performance is the ability of an individual’s body to carry out physical activities efficiently and effectively. Carbohydrates can be the main fuel for various functions, such as physical, cognitive, and cellular activities. The main objective of this experimental study is to determine the effect of the intake of glucose, a form of carbohydrate, on the physical performance of non-athlete students when measured using the Cooper test method of running 2.4 km using a pretest-posttest group research design. The subjects were 30 non-athlete college students. Anthropometric data and physical assessments such as body weight, height, body mass index, body temperature, blood pressure, pulse rate, and blood glucose levels were measured. The length of time measured by the Cooper test method of running 2.4 km for the pre test is 906.7±189.62 seconds. For the post-test, a 6% glucose solution was given to the subjects after a recovery period of 168 hours. The length of time measured by the Cooper test method of running 2.4 km for the post-test is 837.6±173.94 seconds. The comparative data using a t-test shows a p-value of 0.0021 (p<.005), which indicates a significant difference between before and after glucose intake. The subjects ran faster after being given a glucose solution. Therefore, this study concluded that glucose intake significantly affects the physical performance of non-athlete students as measured by the Cooper test method of running 2.4 km.