This study focuses on the electrochemical characterization of Zn-Cu bio-battery cells utilizing electrolytes derived from ambarella peel waste. The primary objectives are to determine the half-cell and full-cell characteristics of these bio-batteries at various concentration ratios and to identify the optimal concentration ratio for maximum performance. Cyclic voltammetry analysis of the half-cells revealed an oxidation peak at 0.5 V vs Ag/AgCl, corresponding to the conversion of uronic acid to aldaric acid. Additionally, two reduction peaks were observed: hydrogen ion reduction to H2 at 0 V vs Ag/AgCl and water reduction at –0.42 V vs Ag/AgCl. The rate-determining step analysis indicated that the redox reactions in the ambarella peel electrolyte solution were surface reactions. The highest rate constant (ks) of 0.722 ± 0.05 s–1 was observed at a 1:2 concentration ratio. This ratio also resulted in the highest battery capacity of 0.0816 mAh and the maximum power density of 16.13 mW/m2. The study concluded that the 1:2 concentration ratio of ambarella peel waste electrolyte solution is optimal, outperforming the 1:1 and 1:3 ratios in terms of battery capacity and power density.