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The aim of this study is 1) to demonstrate the interactions resulting from mixing ethylenediaminetetraacetic acid (EDTA) withsodium hypochlorite (NaOCl) and 2) to monitor the experimental interaction after cold plasma energy transferring mimicking physicochemical activation of solutions. Freshly opened pharmaceutical lavage solutions of 5.25% hypochlorite and 17% EDTA were used in experiments. A mixture of 40 ml was prepared using 5.25% hypochlorite/17% EDTA at a 1:1 ratio. The experimental cold atmospheric-pressure plasma jet (APPJ) treatment was performed using a dielectric barrier discharge (DBD) device for mimicking the activation of lavage solutions. pH measurements of the samples were performed. Spectral changes were analyzed with nuclear magnetic resonance spectroscopy, Fourier-transform infrared spectroscopy, and UV-visible spectrophotometry. N-CH2-COOR and N-CH2-CH2-N were observed in EDTA at 4.65 and 2.90 p/min, respectively. The emergence of new signals at 3.70, 3.07, and 2.91 p/min was observed in the mixture of the EDTA and NaOCl. Decreased peak intensities of EDTA were observed in the APPJ treatment groups at 4.65 and 2.90 p/min with no spectral shift. The main findings are that carbon and nitrogen bonds formed on the EDTA molecule at 2357.08 and 2309.10 cm $^{\mathrm {-1 {}}}$ after oxidation with hypochlorite as middle-narrow and mid-wide, respectively. The molecular structure of EDTA is observed to change after the oxidization by hypochlorite. The newly formed peaks attributed to carbon and nitrogen on the oxidized EDTA molecule disappeared after the experimental APPJ application. The obtained absorbance maxima in the APPJ treatment are in the characteristic region for the $\pi $ - $\pi ^\ast $ transitions. Within the limitations of an in vitro study, the findings of the study demonstrated that the subsequent molecule occurred by the oxidization by hypochlorite or activated hypochlorite is different from each other and also is different from the characteristic structure of EDTA. Experimentally activated hypochlorite by APPJ might have caused the molecular electronic transitions of EDTA. [ABSTRACT FROM AUTHOR]