Nutraceuticals, or functional foods, are foods that have been altered or designed to have certain nutritional or physiological effects. Synbiotic biscuit represents an effort in the development of functional food products. A synbiotic biscuit contains both prebiotic ingredients and probiotic bacteria. Created with Lactobacillus acidophilus and a composite flour mix of sweet potatoes, banana corm, and black soybeans, this synbiotic biscuit offers a synergistic combination of probiotics and prebiotics potentially lowering blood glucose levels. This study investigates the impact of the Glycemic Index (GI), Glycemic Load (GL), and the relationship between the quantity of synbiotic biscuits consumed and the reduction of blood glucose levels in Wistar rats. The descriptive approach was utilized as the experimental method, followed by correlation and regression analysis. The experiment used standard meal consumption as a negative control, metformin 45 mg/kg body weight intake as a positive control, synbiotic biscuit 5 g in 10 ml aquades intake, and synbiotic biscuit 10 g in 10 ml aquades intake were employed in the experiment. The study demonstrated a relationship between the intake of synbiotic cookies and blood glucose levels in Wistar rats. The synbiotic biscuits showed that GI and GL at 65.38 and 13.38, respectively. After 15 days of feeding synbiotic biscuit 10 g in 10 ml per day, body weight increased to 216.33 g, and blood glucose levels decreased to 96.97 mg/dl. Total L. acidophilus was discovered in rat faeces at a concentration of 6.56 log CFU/g.