Fluorescence measurements of peroxynitrite/peroxynitrous acid in cold air plasma treated aqueous solutions.

Qualitative detection of peroxynitrite/peroxynitrous acid (ONOO-/ONOOH) as one of the key bactericidal agents produced in cold air plasma activated aqueous solutions is presented. We examined the use of the 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) fluorescent dye to detect ONOO-/ONOOH in plasma activated non-buffered water (PAW) or buffered solution (PAPB) generated by DC-driven self-pulsed transient spark discharge at atmospheric pressure in ambient air. The diagnostic selectivity of H2DCFDA to reactive oxygen and nitrogen species (RONS) typical of plasma activated aqueous solutions was examined by using various scavengers of RONS. This cross-reactivity study showed the highest sensitivity of the H2DCFDA dye to ONOO-/ONOOH. However, besides ONOO-/ONOOH, H2DCFDA also exhibited sensitivity to hypochlorite anions/hypochlorous acid (OCl-/HOCl), showing that for a selective study it is important to have an idea about the possible constituents in the studied solutions. The sensitivity of H2DCFDA to other RONS even in much higher concentrations was negligible. The presence of nitrites (NO2-) and hydrogen peroxide (H2O2) in PAW led predominantly to the production of peroxynitrous acid with a strong fluorescence response of H2DCFDA in PAW. Plasma treatment of buffered solutions led to the weak response of H2DCFDA. The fluorescence induced in PAW decreased after scavenging individual reactants, namely NO2- and H2O2, as well as by scavenging the product of the peroxynitrite forming reaction, proving that the fluorescence response of H2DCFDA is primarily due to the formation of ONOO-/ONOOH. A chemical kinetics analysis of post-discharge processes and the pseudo-second order reaction between H2O2 and NO2- confirms formation of peroxynitrous acid in PAW with a rate in the order of tens of nM per second. The post-discharge evolution of the ONOOH formation rate was clearly correlated with the parallel detection of ONOO-/ONOOH by fluorescence spectroscopy using the H2DCFDA dye.

Evaluation of Selected Properties of Dielectric Barrier Discharge Plasma Jet.

In the technological processes requiring mild treatment, such as soft materials processing or medical applications, an important role is played by non-equilibrium plasma reactors with dielectric barrier discharge (DBD), that when generated in noble gases allows for the effective treatment of biological material at a low temperature. The aim of this study is to determine the operating parameters of an atmospheric pressure, radio-frequency DBD plasma jet reactor for the precise treatment of biological materials. The tested parameters were the shape of the discharge (its length and volume), current and voltage signals, as well as the power consumed by the reactor for various composition and flow rates of the working gas. To determine the applicability in medicine, the temperature, pH, concentrations of H2O2, NO2- and NO3- and Escherichia coli log reduction in the plasma treated liquids were determined. The obtained results show that for certain operating parameters, a narrow shape of plasma stream can generate significant amounts of H2O2, allowing for the mild decontamination of bacteria at a relatively low power of the system, safe for the treatment of biological materials.

Chemical and Antimicrobial Effects of Air Non-Thermal Plasma Processing of Fresh Apple Juice with Focus on Safety Aspects.

Freshly squeezed apple juice was subjected to air non-thermal plasma treatment to investigate the capability of this processing method to inactivate microorganisms and to evaluate its safety when applied to liquid food products. Two different configurations of a transient spark discharge in ambient air were tested: an electrospray system with the juice flowing directly through the high voltage needle electrode, and a batch system, where the discharge was generated onto the surface of the juice. The key physico-chemical parameters of the juice, such as pH, conductivity, color, transmittance, and Brix degree, did not significantly change upon treatment. The concentration of nitrate ions formed by the plasma was safe, while that of nitrite ions and hydrogen peroxide was initially higher than the safety limits, but decreased within 24 h post treatment. The plasma effect on individual natural components of the juice, such as sugars, organic acids, and polyphenols, treated in water solutions led to their partial or substantial decomposition. However, when these compounds were plasma-treated altogether in the juice, they remained unaffected. The antimicrobial effect of the plasma processing was evaluated via the inoculation of model microorganisms. A stronger (6 log) decontamination was detected for bacteria Escherichia coli with respect to yeast Saccharomyces cerevisiae. Plasma processing led to a substantial extension of the juice shelf-life by up to 26 days if refrigerated, which represents a promising application potential in food technology.

Effects of air transient spark discharge and helium plasma jet on water, bacteria, cells, and biomolecules.

Atmospheric pressure DC-driven self-pulsing transient spark (TS) discharge operated in air and pulse-driven dielectric barrier discharge plasma jet (PJ) operated in helium in contact with water solutions were used for inducing chemical effects in water solutions, and the treatment of bacteria (Escherichia coli), mammalian cells (Vero line normal cells, HeLa line cancerous cells), deoxyribonucleic acid (dsDNA), and protein (bovine serum albumin). Two different methods of water solution supply were used in the TS: water electrode system and water spray system. The effects of both TS systems and the PJ were compared, as well as a direct exposure of the solution to the discharge with an indirect exposure to the discharge activated gas flow. The chemical analysis of water solutions was performed by using colorimetric methods of UV-VIS absorption spectrophotometry. The bactericidal effects of the discharges on bacteria were evaluated by standard microbiological plate count method. Viability, apoptosis and cell cycle were assessed in normal and cancerous cells. Viability of cells was evaluated by trypan blue exclusion test, apoptosis by Annexin V-FITC/propidium iodide assay, and cell cycle progression by propidium iodide/RNase test. The effect of the discharges on deoxyribonucleic acid and protein were evaluated by fluorescence and UV absorption spectroscopy. The results of bacterial and mammalian cell viability, apoptosis, and cell cycle clearly show that cold plasma can inactivate bacteria and selectively target cancerous cells, which is very important for possible future development of new plasma therapeutic strategies in biomedicine. The authors found that all investigated bio-effects were stronger with the air TS discharge than with the He PJ, even in indirect exposure.