Currently, most synthetic-based beverages are hypertonic drinks. In order to maintain osmolality and its ionic potential of 250 m Osm/kg, the possibility of using sugar sweeteners in functional drinks while maintaining their osmolality properties are limited. Hence, the present study used exclusion zone (EZ) water to create an isotonic fruit drink with natural components including high levels of bioactive compounds, along with macro-, micro-, and trace elements, which can be included in an isosmotic cellular health diet. This study uses the pulp and peel of red bananas (Musa acuminata Colla cv. Red) to create isotonic formulations using exclusion zone (EZ) water produced by a self-made device . Eight ingredients with varying weight percentages were combined using a proprietary mixing device. The results were analyzed using Stat-Ease Inc. (USA) Design-Expert 10.0 software. The D-optimal mixture design methodology was used to study the results, revealing that EZ water (90.353 percent), red banana pulp (4.941 percent), red banana peel (0.688 percent), coconut (3.353 percent), isabgol (0.136 percent), propolis (0.024 percent), okra mucilage (0.029 percent), and almond gum (0.477 percent) is the optimized formulation mixture to attain osmolality (285.593). The achieved osmolality was the same as that of serum, a common hydration marker. Further, we evaluated hydration using serum, urine, and saliva samples. The findings revealed that a significant drop in urine volume (1766.45 80.80ml) and the salivary flow rate (0.56 0.05g/min to 0.66 0.04g/min), along with a significant increase in serum [Na+] (1141.59 0.22 mmol/L) and osmolality (288.56 0.61 mmol/kgH2O). A maximum of 1.47 was recorded for the specific gravity of the formulated isotonic beverage. Its micro- and macronutrient composition was as follows: sodium (1.23 mg/100g), magnesium (44.77 mg/100g), calcium (5.59 mg/100g), energy (41.92Kcal/100g), protein (0.17%), soluble dietary fiber (0.87), total sugars (11.73%), carbohydrates (8.18%), vitamin B6 (0.31 mg/100g), and vitamin C (35 mg/100g). The findings indicate that this beverage is unique in aiding rehydration because it provides the right balance of nutrients and hydration for cells to stay healthy.