# Formation of rons including ONOO- and O2 - in physiological buffers exposed to cold atmospheric plasmas

Fanny Girard-Sahun, Vasilica-Adriana Badets, Pauline Lefrançois, Sylvie Blanc, Neso Sojic, Stéphane Arbault, Franck Clement

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Fanny Girard-Sahun, Vasilica-Adriana Badets, Pauline Lefrançois, Sylvie Blanc, Neso Sojic, et al. Formation of rons including ONOO- and O2 - in physiological buffers exposed to cold atmospheric plasmas. 6th International Workshop on Plasma for Cancer Treatment, Apr 2019, Antwerp, Belgium. hal-02183237

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Submitted on 23 Aug 2023

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Distributed under a Creative Commons Attribution 4.0 International License
FORMATION OF RONS INCLUDING ONOO⁻ AND O₂°⁻
IN PHYSIOLOGICAL BUFFERS
EXPOSED TO COLD ATMOSPHERIC PLASMAS

Fanny Girard-Sahun¹, Vasilica Badets², Pauline Lefrançois², Sylvie Blanc¹, Neso Sojic²,
Stéphane Arbault², Franck Clément¹

¹ UPPA, IPREM, CNRS UMR 5254, 2 av Président Angot, Pau, 64000, France

² Univ. BORDEAUX, ISM, CNRS UMR 5255, NSysA Group ENSCBP, Pessac, 33607, France
email: girard-fanny5@orange.fr, stephane.arbault@enscbp.fr, franck.clement@univ-pau.fr

Plasma-liquid interactions are increasingly studied over the last years, within the framework of biomedical applications of cold atmospheric plasmas (CAPs). Among various energy components of CAPs, reactive oxygen and nitrogen species (RONS) play a key role in the reported biological effects [1]. Stable ones (H₂O₂, NO₂⁻, NO₃⁻) remain in the liquid during several hours after CAP treatment and can be detected by different methods [2]. A major challenge consists of the detection and quantification of the unstable and very reactive forms of RONS (such as O₂°⁻, NO° or ONOO⁻).

Herein, the generation of RONS by CAPs in a common physiological buffer (PBS, pH 7.4) was investigated. Plasma device consists of the formation of guided ionizing waves with the control of the gaseous environment. Correlations between plasma gaseous compositions and induced-chemistry in PBS have been studied by electrochemistry and UV-Visible absorption spectroscopy. First, the chemically stable RONS (hour time-scale) were determined for various surrounding environments. Secondly, the unstable RONS (< sec. at physiological pH) were analyzed following two innovative and complementary approaches; the first one consists in stabilizing RONS by switching the pH of PBS to alkaline values, while the second one consists in analyzing RONS generation close to the plasma-liquid interface owing to an *in situ* and in real time electrochemical detection with microelectrodes. Following this dual protocol, we demonstrated the presence and quantified the highly reactive peroxynitrite ONOO⁻, a biologically nitrating and nitrosating species, which ends up in stable NO₂⁻ and NO₃⁻ species, as observed after minutes and hours post-treatment ([2], [3]). Moreover, *in situ* electrochemistry gave information about the liquid chemistry occurring in the first minutes of plasma treatment and allowed the detection of superoxide O₂°⁻ heavily depending on the gaseous environment of the CAPs.

References

[1] D. B. Graves, *Journal of Physics Applied Physics*, **45** (26), 263001 (2012)

[2] F. Girard, V. Badets, S. Blanc, K. Gazeli, L. Marlin, L. Authier, P. Svarnas, N. Sojic, F. Clément, and S. Arbault, *RSC Advances*, **6** (82), 78457–78467 (2016)

[3] F. Girard, M. Peret, N. Dumont, V. Badets, S. Blanc, K. Gazeli, C. Noël, T. Belmonte, L. Marlin, J-P. Cambus, G. Simon, N. Sojic, B. Held, S. Arbault and F. Clément, *Physical Chemistry Chemical Physics*, **20**, 9198-9210 (2018)