The utilization of Cold Plasma (CP) technology for decontamination and disinfection has garnered considerable attention across diverse industries. This study aims to investigate the interaction between pH and electrical conductivity (EC) (μS/cm) in Cold Plasma-Activated Water (CPAW) enriched with metal cations and its potential as an antifungal agent against two Penicillium (P.) mold strains. The investigation focuses on elucidating the augmented chemical interactions induced by plasma between radicals, charged particles, and microorganisms' cell membranes within an aqueous environment. Our findings demonstrate a positive correlation between the inactivation potential of CPAW (operating at 10 kV voltage, 2.5 kHz high frequency, and 500 mA current intensity) and pH and EC(μS/cm) values. Notably, the relative chemical reactivity and solubility of calcium oxide emerge as significant factors, highlighting the pronounced link between P. Italicum and Plasma-Activated Water containing Copper cations (CPAW + Cu 2+ ) (p < 0.05). Our study distinctly emphasizes (1) the substantial impact of both activated water type and mold species on CFU/mL values (p < 0.05); (2) the mold-specific effect of activated water on CFU/mL; and (3) the noteworthy EC(μS/cm) enhancement and pH decrease with prolonged activation time, attaining statistical significance (p < 0.01).