Quantifying the concentration and penetration depth of long-lived RONS in plasma-activated water by UV absorption spectroscopy

Reactive oxygen and reactive nitrogen species (RONS) are believed to play a key role in biomedical applications, which means that RONS must reach the target tissue to produce a therapeutic effect. Existing methods (electron spin spectrometry and microplate reading) to determine the RONS concentration are not suitable for experimental real-time measurements because they require adding an indicating reagent to the plasma-treated medium, which may alter the chemical composition of the medium. In this paper, we propose a method to measure the long-lived RONS concentration in plasma-activated water (PAW) by using UV absorption spectroscopy. Based on an analysis and fit of the absorption spectra of standard solutions (H2O2, NaNO2, and NaNO3), we propose a detailed fitting procedure that allows us to calculate the concentrations of simplex H2O2, NO2, and NO3. The results show that the pH and the cross reactivity between RONS in PAW correlate strongly with the absorption spectra. To confirm the accuracy of the calculations, we also use a microplate reader and add chemical reagents to measure the concentrations of H2O2, NO2, and NO3. The results show that the concentrations calculated by the proposed fitting method are relatively accurate and that the error range is acceptable. Additionally, the time-dependent diffusion of RONS in PAW is measured and analyzed at different depths in the PAW. This fitting approach constitutes a nonintrusive approach to measure RONS at different depths in PAW. (C) 2019 Author(s).