Purpose: This study aimed to verify the impact of the time that the sitting posture is maintained on changes in muscle performance and head alignment. Methods: The subjects of this study were healthy adults aged between 20 and 30 years. Ninety-three subjects participated in this study (male: 57, female: 36). The subjects were divided into a one-hour group, a two-hour group, and a three-hour group. All the subjects adjusted the height of their chair to a comfortable position and then seat for one, two, or over three hours. Both prior to and after the experiment, the muscle performance (muscle strength, endurance, and flexibility) of the subjects was measured. Results: In the 2-hours and 3-hour group, muscle strength, flexibility and endurance reduced significantly before and after the study, and the head alignment significantly changed. Conclusion: Thease findings showed that sitting continuously for longer than two hours decreases muscle strength, endurance, and flexibility. A flexed posture affects the muscle performance of the lumbar region and may result in problematic postures, such as a forward head position.