# Formation of reactive oxygen and nitrogen species including peroxynitrite in physiological buffer exposed to cold atmospheric plasma

Fanny Girard, Vasilica-Adriana Badets, Sylvie Blanc, Néso Sojic, Franck Clement, Stéphane Arbault

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## To cite this version:

Fanny Girard, Vasilica-Adriana Badets, Sylvie Blanc, Néso Sojic, Franck Clement, et al.. Formation of reactive oxygen and nitrogen species including peroxynitrite in physiological buffer exposed to cold atmospheric plasma. 6th International Conference on Plasma Medicine, Sep 2016, Bratislava, Slovakia. hal-05367809

HAL Id: hal-05367809

https://univ-pau.hal.science/hal-05367809v1

Submitted on 16 Nov 2025

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Distributed under a Creative Commons Attribution 4.0 International License
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UNIVERSITÉ
DE PAU ET DES
PAYS DE L'ADOUR

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IPREM

Institut des sciences analytiques
et de physique-chimie
pour l'environnement et les milieux

Université
de BORDEAUX

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NSysA
NanoSystèmes Analytiques

# Formation of reactive oxygen and nitrogen species including peroxynitrite in physiological buffer exposed to cold atmospheric plasma

Fanny Girard,ab Vasilica Badets,b Sylvie Blanc,a Neso Sojic,b Franck Clémenta

and Stéphane Arbaultb

fanny.girard@univ-pau.fr

franck.clement@univ-pau.fr

stephane.arbault@enscbp.fr

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icpmb

6th International
Conference on
Plasma Medicine

Bratislava - Slovakia
September 4-9. 2016

AGENCE NATIONALE DE LA RECHERCHE
ANR
PlasmaRegen

<footnote>a UPPA, IPREM, CNRS UMR 5254, 2 avenue Président Angot, 64000 Pau, France.</footnote>

<footnote>bUniv. BORDEAUX, ISM, CNRS UMR 5255, NSysA group, ENSCBP, 16 avenue Pey Berland, 33607 Pessac, France.</footnote>
Context

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Outline

**Physical parameters**

Gas, Voltage, frequency,
duty cycle...

Plasma generation
and
characterization

**Plasmas**

Physico-chemical components
(UV, e.m. fields, gaseous RONS)

Plasma-liquid
interactions

**Liquid**

pH, σ, RONS, evaporation
# Plasma setup

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« Guided ionization wave »

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7.5 kV_10 kHz_duty cycle = 1 %
# 1) Plasma generation and characterization

## Experimental setup 1

### Working gas

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➤ PBS -- (10 mM) pH 7.4

### Working gas : He/1%N₂

<figure><img src="image_14.png" /></figure>

7.5 kV
10 kHz
duty cycle = 1 %
2 slm

Air
environment

<figure><img src="image_15.png" /></figure>

No control of the
surrounding
atmosphere
# 1) Plasma generation and characterization
## Optical emissive spectroscopy

<figure><img src="image_16.png" /></figure>

<figure><img src="image_17.png" /></figure>

► **Rotational structures of OH and N₂(SPS) → Tgas ≈ 300 K***

<footnote>*From Gazeli et al., Journal of Applied Physics 117, 093302. 2015</footnote>
## 2) Plasma-liquid interactions

### Experimental protocol

Excited chemical species in the plasma

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What effects on liquid chemistry ?

### 1) Plasma treatment

- Various plasmas

- Various exposure times

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### 2) Liquid analysis

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UV-visible absorption spectroscopy

Electrochemistry

POST-TRAITEMENT
Analysis

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<figure><img src="image_23.png" /><figcaption>7.5 kV
10 kHz
duty cycle = 1 %</figcaption></figure>

Evaporation phenomenon *

<figure><img src="image_24.png" /></figure>

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Evaporation depends on working gas composition
<figure><img src="image_26.png" /><figcaption>He/1%N₂</figcaption></figure>

UV-visible absorption spectroscopy

<figure><img src="image_27.png" /></figure>

Electrochemistry

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Stable species :

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Validated with 2
techniques

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Effects on
pH ?
7.5 kV
10 kHz
duty cycle = 1 %

pH

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* Acidification

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* Hypothesis : HNO₂ and HNO₃
## 2) Plasma-liquid interactions

### RONS

What strategy to detect
unstable oxygen and nitrogen
species (RONS) in liquids ?

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Switch to alkaline values

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Treatment

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UV-visible absorption
spectroscopy
## 2) Plasma-liquid interactions

### RONS

### PBS pH 12, UV-visible absorption spectroscopy *

* 7.5 kV
* 10 kHz
* duty cycle = 1 %
* 2 slm

<figure><img src="image_38.png" /><figcaption>He/1%N₂</figcaption></figure>

<figure><img src="image_39.png" /></figure>

* ✓ ONOO⁻ stabilized at pH 12

* ✓ [ONOO⁻] ≈ 50 µM (10 minutes of treatment)

<footnote>*Girard et al., RSC Adv., 2016, 6, 78457–78467</footnote>
#### Influence of the surrounding atmosphere on chemical species ?

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##### Shielding gas device

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Switch to alkaline values : PBS pH 12

7.5 kV
10 kHz
duty cycle = 1 %

<figure><img src="image_44.png" /><figcaption>ONOO⁻ quantification_treatment 10 min</figcaption></figure>

<figure><img src="image_45.png" /></figure>

✓ [ONOO⁻]max for 20% O₂/80% N₂ in the shielding gas (“air shielding”)

✓ [ONOO⁻]max ≈ 25 µM
7.5 kV
10 kHz
duty cycle = 1 %

PBS pH 7.4

<figure><img src="image_46.png" /></figure>

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* ✓ $[NO_2^-]_{max}$ for the "air shielding" (20% O₂/80% N₂)

* ✓ $[H_2O_2]_{max}$ for 100% N₂ in the shielding gas
## 3) Shielding device

### Experimental setup 2 : evaporation

### Evaporation phenomenon

#### Setup 1

<figure><img src="image_48.png" /></figure>

No shielding gas
device

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Evaporation depends on the
working gas composition

#### Setup 2

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Working gas : He
+
Shielding gas

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Evaporation controlled
Conclusions and questionings...

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Thank you for your
attention

➤ Particular thanks to Laurent Marlin, Natacha Dumont, Mathieu Peret and Kristaq Gazeli for their respective contribution

fanny.girard@univ-pau.fr
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#### Electrochemistry_Cyclic voltammetry

WE : Pt Ø20 µm microelectrode + Pt black deposit (30 µC -60mV)

REF : Ag/AgCl

CE : Pt

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#### UV-visible absorption spectroscopy

5 cm cuve length, T°$_{amb}$, PBS

<figure><img src="image_58.png" /></figure>
7.5 kV
10 kHz
duty cycle = 1 %

Switch to alkaline values : PBS pH 12 *

UV-visible absorption spectroscopy

<figure><img src="image_59.png" /><figcaption>He/1%N₂</figcaption></figure>

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➤ 1st Hypothesis : hypochlorite ClO⁻ (ε₂₉₂nm = 365 M⁻¹·cm⁻¹)

➤ 2nd Hypothesis : ONOO⁻ (ε₃₀₂nm = 1670 M⁻¹·cm⁻¹)

<footnote>*Girard et al., RSC Adv., 2016, 6, 78457–78467</footnote>
7.5 kV
10 kHz
duty cycle = 1 %

Switch to alkaline values : PBS pH 12 *

UV-visible absorption spectroscopy

He/1%N₂

<figure><img src="image_61.png" /></figure>

Beer-Lambert law :

$$A_{\lambda} = \sum_{i} \varepsilon_{\lambda,i} |C_{i}|$$

<figure><img src="image_62.png" /></figure>

* ✓ ONOO⁻ hypothesis validated

* ✓ [ONOO⁻] ≈ 50 µM (10 minutes of treatment)
<figure><img src="image_63.png" /><figcaption>pH-nitrite correlation</figcaption></figure>

<figure><img src="image_64.png" /></figure>

* Acidification

<figure><img src="image_65.png" /></figure>

* Hypothesis : HNO₂