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This study compared the sensing characteristics of ZnO (ZO) treated with ammonia (NH₃) plasma for 1 min, 3 min, and 6 min, under the EIS structure. The measurement results revealed that, after 3 min of NH₃ plasma treatment, the Mg-doped ZnO (MZO) sensing film had a high hydrogen ion sensitivity, linearity, hysteresis, and drift rate of 53.82 mV/pH, 99.04%, 2.52 mV, and 1.75 mV/h, respectively. The sensing film was used with sodium and potassium ion solutions, and it performed satisfactorily in sensing hydrogen ions. Additionally, we investigated the biomedical sensing properties of Mg-doped ZnO (MZO) sensing film with regard to urea, creatinine, and glucose solutions and found that the Mg-doped ZnO (MZO) sensing film treated with NH₃ plasma for 3 min had the best properties for sensing urea, creatinine, and glucose. Specifically, with glucose, the sensing film achieved the best linearity and sensitivity and of 97.87% and 10.73 mV/mM, respectively. The results revealed that the sensing characteristics varied with the processing environment and are useful in the developing biomedical sensing applications with different sensing elements.