Aim: This study aimed to evaluate the physiological effects of intermittent fasting (IF) on glucose homeostasis, insulin sensitivity, and metabolic parameters in adults over a 12-week period. Materials and Methods: A prospective, randomized controlled trial was conducted with 120 adult participants (aged 25–60 years) recruited from the outpatient department of physiology. Participants were randomly assigned to either the IF group (n = 60), following a 16:8 fasting protocol, or the control group (n = 60), maintaining a regular eating pattern. Anthropometric, biochemical, and metabolic parameters were assessed at baseline, week 6, and week 12. Fasting plasma glucose (FPG), postprandial glucose (PPG), HbA1c, fasting insulin, HOMA-IR, lipid profile, and blood pressure were measured. Results: The IF group showed greater reductions in body weight (6.72%), BMI (7.03%), and waist circumference (4.08 cm) compared to the control group. FPG and PPG decreased by 12.21% and 17.65%, respectively, in the IF group, whereas the control group showed smaller reductions (3.66% and 4.36%). HbA1c declined by 8.68% in the IF group versus 2.31% in the control group. Fasting insulin levels decreased by 33.12% in the IF group, while the control group showed a 12.46% reduction. Improvements in total cholesterol (10.45%), LDL (10.88%), and HDL (12.50%) were observed in the IF group, along with significant reductions in systolic (9.00%) and diastolic (7.59%) blood pressure. Despite these improvements, p-values for most parameters were above 0.05, indicating trends rather than statistically significant differences. Conclusion: Intermittent fasting demonstrated positive effects on metabolic health, including weight loss, improved glycemic control, insulin sensitivity, and lipid profile regulation. Although the results were not statistically significant, the findings align with previous research supporting IF as an effective dietary strategy for metabolic disorders. Longer-duration studies with larger sample sizes are needed to establish its clinical relevance.