Plasma-activated Water Research Articles

Effect of Plasma-Activated Water (PAW) on the Postharvest Quality of Shepherd's Purse (Capsella bursa-pastoris).

Plasma-activated water (PAW) treatment is an effective technique for the quality retention of fresh vegetables with cold atmospheric plasma using controllable parameters. This study investigated the effect of PAW on the postharvest quality of shepherd's purse (Capsella bursa-pastoris). The results displayed that PAW treatment with an activation time of 5, 10, 15, and 20 min reduced the yellowing rate and weight loss of the shepherd's purse during 9 days of storage. Compared with untreated samples, PAW treatment at different times reduced the number of total bacteria, coliform, yeast, and mold by 0.18-0.94, 0.59-0.97, 0.90-1.18, and 1.03-1.17 Log CFU/g after 9 days of storage, respectively. Additionally, the treatments with PAW-5 and PAW-10 better preserved ascorbic acid, chlorophyll, total phenol, and total flavonoid contents. They also maintained the higher antioxidant and CAT activity and inhibited the formation of terpenes, alcohols, and nitrogen oxide compounds of the shepherd's purse at the end of storage. The microstructural result illustrated that the cells of the shepherd's purse treated with PAW-5 and PAW-10 were relatively intact, with a small intercellular space after storage. This study demonstrated that PAW treatment effectively improved the postharvest quality of shepherd's purse.

Seed priming with gas plasma-activated water in Ethiopia’s “orphan” crop tef (Eragrostis tef)

Main conclusionSeed priming with gas plasma-activated water results in an increased ageing resilience in Eragrostis tef grains compared to a conventional hydropriming protocol.Tef (Eragrostis tef) is a cereal grass and a major staple crop of Ethiopia and Eritrea. Despite its significant importance in terms of production, consumption, and cash crop value, tef has been understudied and its productivity is low. In this study, tef grains have undergone different priming treatments to enhance seed vigour and seedling performance. A conventional hydropriming and a novel additive priming technology with gas plasma-activated water (GPAW) have been used and tef grains were then subjected to germination performance assays and accelerated ageing. Tef priming increases the germination speed and vigour of the grains. Priming with GPAW retained the seed storage potential after ageing, therefore, presenting an innovative environmental-friendly seed technology with the prospect to address variable weather conditions and ultimately food insecurity. Seed technology opens new possibilities to increase productivity of tef crop farming to achieve a secure and resilient tef food system and economic growth in Ethiopia by sustainable intensification of agriculture beyond breeding.

Implementing water recirculation in a novel portable plasma-activated water reactor enhances antimicrobial effect against Escherichia coli

The need for sustainable and reliable decontamination methods is driven by concerns regarding antibiotic resistance, as well as environmental and cost-efficiency challenges associated with traditional methods. Plasma-activated water (PAW) holds significant promise as an innovative and eco-friendly decontamination method. Nevertheless, prior to industrial implementation of PAW-based decontamination devices, a deep understanding of the bacterial inactivation mechanisms and its interplay with PAW chemical composition is required. Advancing in this field requires interdisciplinary and collaborative research using standardized practices with cost-effective and shareable PAW reactors that are still missing today. Here, to this end, a portable PAW reactor, featuring a surface dielectric barrier discharge (SDBD) that operates in air, is presented. The gaseous and aqueous phases were chemically characterized by Fourier transform infrared (FTIR) absorption spectroscopy and vis-spectrophotometry, respectively. To assess PAW antimicrobial efficacy Escherichia coli was employed as a model organism on six different PAW samples for three different treatment times. The significance of water recirculation in controlling the chemical composition of PAW, altering the nitrogen species balance in favor of NO2−, is demonstrated. This chemical modification consequently enhanced the antimicrobial effectiveness of PAW, reaching a log reduction of ∼6. Exposing E. coli to equivalent concentrations of RONS to those found in PAW attained similar log-reductions, indicating that acidified nitrites are key to PAW antimicrobial activity. Through an innovative and portable design, this study illustrates the crucial role of water recirculation in tailoring PAW composition, leading to improved decontamination capabilities and establishing a key parameter for further optimization of PAW production.

Iron nanoparticle coating and plasma-activated water cleaning as a biofouling removal method for reverse osmosis membranes suitable for desalination

Desalination provides crucial freshwater, but biofouling on reverse osmosis (RO) membranes is a major challenge due to hard-to-remove biofilms. Conventional treatments are often ineffective, prompting a shift to coatings with nanoparticles like copper, silver, and iron to combat fouling. This study demonstrates a unique combination of coating RO membranes with iron nanoparticles (FeNPs) and treatment with plasma-activated water (PAW) for the removal of biofilm formation. Pure and unbound FeNPs showed no significant inhibition of Escherichia coli biofilms. However, the attachment of FeNPs to RO membranes led to a decrease in biofilm growth on the membrane, and an additional PAW treatment was shown to significantly inhibit mature biofilms. SEM imaging revealed a higher density and uniform distribution of FeNPs on the RO membranes after PAW treatment. Zeta potential measurements showed positively charged FeNPs in PAW, enhancing their attraction with the negative membrane surfaces. The coated membrane using PAW solution with FeNPs exhibited no degradation in terms of pure water flux, saline water flux, and salt rejection compared to the immersion process in DI with FeNPs.

Falling liquid droplets discharge

This paper reports the phenomenon of two plasma segments forming when a water droplet descends, one between the upper part of the droplet and the outlet tube and the other between the lower part of the droplet and the water surface in the container. The study reveals that as the water droplet descends, the length of the upper plasma gradually increases, while the length of the lower plasma decreases until the lower plasma disappears upon contact between the water droplet and the water in the container. The study finds that the rotational temperature of this plasma reaches 2100 K, with an electron density of 1014 cm−3. Furthermore, it is intriguing to note that the descent speed of the droplet is significantly greater than that of a freely falling droplet. Further research indicates that this is due to an instant water channel explosion-induced downward impact on the droplet caused by plasma generation at the upper part of the droplet. The advantage of this device lies in the fact that the plasma only comes into direct contact with water, without any contact with metal electrodes, thus eliminating the issue of electrode corrosion. Furthermore, since the water is in dynamic flow, it facilitates the dissolution of reactive gaseous components into the water, making it suitable for applications related to plasma-activated water and similar purposes.

Chlorpyrifos Pesticide Removal from Black Grapes Using Plasma-Activated Water Produced by Plasma Bubbling Technology

This study is aimed at evaluating the ability of plasma-activated water (PAW) to reduce the chlorpyrifos pesticide residue in black grapes. Raw black grapes were spiked with a known concentration (10, 15, and 20 ppm) of chlorpyrifos (20% EC) and bubbled with 120 V air plasma at three different time intervals (5, 10, and 15 min). GC-MS analysis of these plasma-bubbled grapes showed up to 65.25% reduction (20 ppm sample) in chlorpyrifos content after 15 min of treatment. However, the treatment also reduced the grape juice’s ascorbic acid (19.97±2.69 to 9.15±2.02 mg/ml), antioxidant scavenging activity (77.42±2.97 to 53.30±4.77%), total flavonoids (3.00±0.53 to 2.61±0.57 mg QE/ml), total soluble solids (14.23±1.96 to 11.95±1.86°), total suspended solids (1.95±0.16 to 1.80±0.03 g/l), and turbidity (246.63±11.42 to 224.1±24.85 NTU). Meanwhile, other physicochemical attributes such as pH, titrable acidity, total phenol content, color index, and texture values had slight changes after plasma bubbling. Thus, plasma bubbling proved to be an effective method to remove the chlorpyrifos pesticide present in grapes, and the techniques also preserve the quality of the commodity.

Combined of plasma-activated water and dielectric barrier discharge atmospheric cold plasma treatment improves the characteristic flavor of Asian sea bass (Lates calcarifer) through facilitating lipid oxidation

To explore the combination effects of plasma-activated water and dielectric barrier discharge (PAW-DBD) cold plasma treatment on the formation of volatile flavor and lipid oxidation in Asian sea bass (ASB), the volatile flavor compounds and lipid profiles were characterized by gas chromatography-ion mobility spectrometry and LC-MS-based lipidomics analyses. In total, 38 volatile flavor compound types were identified, and the PAW-DBD group showed the most kinds of volatile components with a significant (p < 0.05) higher content in aldehydes, ketones, and alcohols. A total of 1500 lipids was detected in lipidomics analysis, phosphatidylcholine was the most followed by triglyceride. The total saturated fatty acids content in PAW-DBD group increased by 105.02 μg/g, while the total content of unsaturated fatty acids decreased by 275.36 μg/g. It can be concluded that the PAW-DBD processing increased both the types and amounts of the volatile flavor in ASB and promoted lipid oxidation by altering lipid profiles.

Atmospheric non-thermal plasma inactivation of Ascosphaera apis, the causative agent of chalkbrood disease in honeybee

Ascosphaera apis is a worldwide pathogenic fungi of honeybees that can cause a decline in bee populations. In this study, we investigated the antifungal activity of non-thermal plasma on fungal growth. Spore inactivation after exposure to gas plasma by liquid phase and plasma activated water (PAW) and pathogenicity of A. apis in vivo were also examined. The results demonstrated that the mycelial growth of fungi was completely inhibited after argon plasma treatment. Both gas plasma and PAW exposures resulted in a significant decrease of A. apis spore numbers, maximum reduction of 1.71 and 3.18-fold, respectively. Germinated fungal spores on potato dextrose agar were also reduced after plasma treatment. SEM analysis revealed a disruption in the morphological structure of the fungal spores. The pathogenicity of A. apis on honeybee larvae was decreased after spores treated by gas plasma and PAW with a disease inhibition of 63.61 ± 7.28% and 58.27 ± 5.87%, respectively after 7 days of cultivation. Chalkbrood in honey bees have limited control options and our findings are encouraging. Here, we demonstrate a possible alternative control method using non-thermal plasma for chalkbrood disease in honeybees.

Effect of cold plasma treatment and plasma‐activated water on physicochemical and structural properties of starch: A green and novel approach for environmental sustainability

AbstractStarch is a crucial part of the human dietary regimen. Still, its restricted water solubility, low viscosity, and weak heat stability are unfavorable physicochemical characteristics that restrict its use in food applications. So, in the 21st century, green and novel technologies such as cold plasma technology (CPT) and plasma‐activated water (PAW) have been employed for starch modification to improve the starch's physicochemical, digestibility, functional, structural, and thermal properties. This article provided an in‐depth evaluation of the use of cold plasma intervention in starch systems. Moreover, starch was modified due to cross‐linking, plasma etching, and depolymerization mechanisms mediated by plasma species. We may conclude that CPT and PAW are substitute methods for changing starch characteristics.

Development of Plasma-Activated Water (PAW) System: Molecular Dynamics Simulation and Experimental Study on Almond Aflatoxins

Almonds play a significant role in Iran’s economy, and factors that threaten their market, such as aflatoxins, need careful consideration. Plasma-activated water (PAW) is a new method that has antioxidant activity and can eliminate toxins and microbial agents, making it a suitable solution for removing aflatoxins from almonds. In a study, the continuous PAW production system was used to control and remove almond aflatoxins, and molecular dynamics simulation was employed to evaluate the impact of PAW on aflatoxin B1. The study found that reducing the flow rate of plasma-treated water had the greatest effect on reducing aflatoxin concentration, followed by PAW application time, the dose of inoculated toxin, and the air/gas mixture ratio. The use of PAW reduced aflatoxin concentration in almonds by 12% to 56.8%. The simulation results suggested that PAW can affect the structure of aflatoxin B1, change and destroy its activity, and reduce its toxicity. Among the free radicals tested, NO3− was found to be the most effective in degrading aflatoxin B1. This study demonstrates the potential of PAW as a method to remove aflatoxins and enhance the safety of almonds.

Disinfection Efficacy of Electrohydraulic Discharge Plasma against Xanthomonas campestris pv campestris: A Sustainable Seed Treatment Approach.

The prevalence of plant diseases caused by pathogens such as Xanthomonas campestris pv campestris (Xcc) poses a significant challenge to sustainable agriculture, necessitating the development of effective and eco-friendly disinfection methods. In this study, we investigated the efficacy of electrohydraulic discharge plasma (EHDP) as a promising alternative for disinfection against Xcc, a pathogen responsible for black rot in cruciferous vegetables. Unlike conventional gas-phase plasma, EHDP introduces two pivotal components: gas-liquid interface plasma (GLIP) and its consequential byproduct, plasma-activated water (PAW). While GLIP enables dual-phase production of reactive oxygen and nitrogen species (RONS), PAW is a reservoir of liquid-phase long-lived RONS, thereby enhancing its bactericidal efficacy. In our evaluations, we tested EHDP-induced GLIP and EHDP-induced PAW against Xcc cells in both in vitro (Xcc suspension) and in vivo (Xcc-inoculated cabbage seeds) settings, achieving noteworthy results. Within 15 min, these methods eliminated ∼98% of the Xcc cells in suspension. For in vivo assessments, nontreated seeds exhibited an infection rate of 98%. In contrast, both EHDP treatments showed a significant reduction, with ∼60% fewer seeds infected while maintaining ∼90% germination rate. In addition, the liquid-phase RONS in EHDP-PAW may enhance seed vigor with a faster germination rate within the initial 5 days. Remarkably, around 90% of EHDP-PAW-treated seeds yielded healthy seedlings, indicating dual benefits in bacterial suppression and seed growth stimulation. In contrast, the percentage of healthy seedlings from nontreated, Xcc-inoculated seeds was approximately 70%. Our research demonstrates the feasibility of using eco-friendly EHDP in the seed disinfection process.

Study on the thawing characteristics of beef in plasma-activated water

Plasma-activated water (PAW) thawing and traditional water thawing were used to study the thawing characteristics and physicochemical properties of beef, and infrared spectroscopy, scanning electron microscopy and other technologies were used as detection methods. The activation times of plasma-activated water (PAW) were 20, 40, 60, 80 and 100 min, respectively. The results showed that, compared with traditional water thawing, the PAW thawing treatment could reduce the total number of bacteria in the meat by 0.5 log, and the medium itself had a more significant bactericidal effect of up to 1.7–2.6 log, and had no adverse effect on the color or pH of the beef. Moreover, PAW maintained the stability of beef fiber, improved the water-holding capacity, inhibited lipid oxidation, and reduced the loss of soluble substances such as proteins. Therefore, PAW thawing is a promising beef thawing technology that provides practical guidance and methods for the wider application of PAW in the field of beef thawing. Industrial relevanceThe thawing characteristics and physicochemical properties of beef by plasma-activated water (PAW) were investigated. It was found that PAW had a positive effect on decontamination bacteria, improving the water-holding capacity of beef, inhibiting lipid oxidation and maintaining the fiber structure of beef. The potential application of PAW in the thawing of frozen meat products was highlighted.

Cold plasma treatment boosts barley germination and seedling vigor: Insights into soluble sugar, starch, and protein modifications

This study investigates the impact of three cold plasma treatments on barley seed germination: direct treatment of dry seeds (DDS), direct treatment of water-soaked seeds (DWS), and indirect treatment of seeds using plasma-activated water (IPAW). Our findings reveal that while DDS maintained the seed's imbibition structures, DWS significantly reduced water uptake. Notably, DWS-treated seeds exhibited a substantial increase in moisture content, potentially leading to an oxygen-deficient environment within the seed. This condition appeared to hinder germination, contrasting with the beneficial effects observed in DDS-treated seeds. Analyses of surface hydrophilicity, phenolic compounds, carbohydrate composition, protein storage and seedling growth were conducted. Interestingly, cold plasma treatment enhanced seed surface hydrophilicity, as evidenced by a decrease in water contact angles, particularly for DWS. We observed specific reductions in flavonoids and total phenolics, alongside shifts in carbohydrate composition, including increased sugar content up to 15.1% (DDS) and 13.5% (DWS), accompanied by a decrease in starch content to 189.00 ± 7.86 mg/g FM for DWS. Protein storage assessments revealed declines in albumin, globulin and prolamin fractions post-CP exposure. Finally, DDS and IPAW treatments markedly improved seedling shoot growth (from two to three-fold), underscoring the potential of these treatments in enhancing barley germination.

Ultrasound-assisted activation of PAW residual radicals in the concurrent elimination of ARB and ARGs: Process efficiency, mechanism and implication

Methicillin-resistant Staphylococcus aureus (MRSA) and antibiotic resistance gene (mecA) are considered emerging contaminants that have been extensively identified in aqueous environments due to their high resistance to disinfectants. Herein, we report for the first time the inactivation of MRSA through a sequential application of plasma-activated water and ultrasound (PAW-SU). While the impact of PAW alone on bacterial culturability and mecA integrity was negligible, the combined PAW-SU treatment outperformed the inactivation method involving PAW with direct US (PAW-DU). It exhibited a shorter lag phase and a higher maximum inactivation rate constant. We numerically demonstrated that sonolysis processes induced by external ultrasound sources considerably improved the intracellular and extracellular reactive oxygen and nitrogen species (RONS) of MRSA in PAW system, thereby increasing the PAW reactivity and applicability. The highest inactivation efficiency of PAW-SU can be attributed to the synergistic effect of enhanced intracellular RONS (i.e., peroxynitrite and singlet oxygen) and membrane disruption induced by US, resulting in irreparable oxidative damage to the Save our Soul response and energy metabolic system. In particular, the elevated arginine metabolites in MRSA cells were primarily responsible for the formation of the viable but nonculturable state (VBNC) bacteria associated with high resistance to PAW system. Surprisingly, a decrease in arginine synthesis capacity was observed to expedite the flow of the tricarboxylic acid cycle, accelerating the detrimental effects of PAW-SU on intracellular components in VBNC bacteria. Finally, the limited regrowth potential of injured MRSA caused by cyclic inactivation of PAW-SU process was further confirmed in wastewater from meat and dairy models and therefore, advocates the possible real-world applications.

Influence of Plasma-Activated Water on Foliar and Fruit Micronutrient Content and Plant Protection Efficiency

Foliar fertilizing is very important to supply apple plants with calcium and micronutrients. The most cost-effective approach to this is the application of the fertilizers in tank mixtures with plant protection chemicals. Plasma-activated water (PAW) has great potential for the use in the agriculture. We used two type of PAWs, PAW1 (made using underwater electrical discharge in an aqueous KNO3 solution and includes reactive nitrogen species and platinum nanoparticles) and PAW2 (made using a plasma torch with nitrogen gas makeup and contains reactive nitrogen species but not metals). We studied the impact of two PAW types on the contents of Ca, B, Mn, Fe, and Co in leaves and Ca, Mn, Fe, Zn, and Mo in fruits sprayed with tank mixtures containing the fertilizers. We also tested the efficiency of PAW in the control of apple scab when applied as tank mixtures with plant protection chemicals. The application of the PAWs significantly increased foliar Ca when the PAW was mixed with Ca-containing formulations (spraying PAW1 containing Ca increased leaf Ca by up to 21%, and PAW2 up by to 9% compared to Ca spraying without PAW). The largest fruit Ca increase was in the variant treated with PAW1 with a micronutrient spraying program (up to 143%). The PAW treatments enhanced the baseline mineral contents of the plants even when they were not sprayed with the nutrients. PAW1 mainly increased the nutrient contents of the apple fruits. PAWs have proven to be efficient for the control of apple scab, thereby reducing the demand for fungicides. The scab damage to the leaves and fruits was similar in plants treated with PAWs without fungicides (1.7–1.9% on the leaves and 1.6–1.8% on the fruits) compared to the conventional chemical scab control (0.9% leaves and 0.6% fruits) and was significantly lower than in the untreated control (9.3% on leaves and 11.9 on fruits).

Cold atmospheric pressure plasma as eco-friendly technology prolonged the vase life and improved the quality of cut rose flowers

Cold atmospheric pressure plasma is a relatively new technology characterized by safety and high efficiency, that is increasingly attracting attention in agriculture and food industries especially in post-harvest technology. This experiment addresses the potential use of plasma-activated water (PAW) for increment of cut rose flower longevity and quality. A complete randomized design was used in this study, and each treatment had three replicates with six flowers in each replicate. Based on the findings of this study, it seems that PAW application as a preservation solution improves plant water status during postharvest storage and reducing the electrolyte leakage and maintaining the relative water content (RWC). Data from the experiment showed that the plasma activated water decreased the malondialdehyde content and maintains the structure and integrity of the cell membrane through the activation of enzymatic and non-enzymatic antioxidant systems and phenolic compounds synthesis ultimately leading to postharvest quality and prolonged vase life of cut rose flowers. As PAW is more cost-effective than many other preservatives and is eco- friendly with fewer side effects, so it could be used in preservative solutions as an alternative.

Effect of air, O2, Ar, and N2 plasma‐activated water on mildewing activity of moldy pathogen of Gannan navel oranges

AbstractIn this study, the physicochemical properties of plasma‐activated water (PAW) prepared from air, , , and plasma and the trend of the concentration of reactive oxygen and nitrogen species (RONS) with respect to time were investigated and related to the effect of PAW on navel orange mold. The results showed that PAW had a strong lethal effect on navel orange mold and could effectively inhibit the mildewing activity of pathogenic spores. In addition, PAW is effective in reducing weight loss and decay of navel oranges during storage. This study suggests the potential application of PAW to prevent fungal infections during storage of agricultural products.

Plasma Activated Water Solution as an Efficient Approach for Antiseptic and Wound Healing Applications: An Animal Model

The current investigation was aimed at identifying the potential of plasma activated water as an antiseptic and wound-healing solution. The simplicity and affordability of the indirect approach of plasma therapies led us to evaluate rinsing of wounds via plasma activated water (PAW) and compare to the direct approach of plasma jet. Although previous studies have shown the effectiveness of plasma-activated water in wound healing and disinfection, this study was conducted to investigate two PAWs with various compositions in wound management. In this regard, PAWs with two different treatment times were produced and used for rinsing of wounds. H&lt;sub&gt;2&lt;/sub&gt;O&lt;sub&gt;2&lt;/sub&gt; and NO&lt;sub&gt;2&lt;/sub&gt;- concentrations as well as acidity values of PAW were measured. Wound healing was investigated to find an efficient solution using Masson trichrome and hematoxylin and eosin staining. The antibacterial effect and wound closure rate of the directly applied plasma jet were significantly improved compared to the PAW treatments. The results exhibited an increase in collagen expression, resulting in the best rate of wound closure. Thus, our results suggested that plasma jet treatment and PAW rinsing of wounds are effective methods without side effects in promoting wound healing.

Field efficacy of Gliding Arc Plasma Activated Water and natural products on tomato (Solanum lycopersicum L.) late blight [Phytophthora infestans (Mont.) de Bary] severity

Abstract This study aimed to evaluate the field efficacy of plasma-activated water (PAW) as a novel technology in agriculture together with vinegar, sodium bicarbonate, and plant aqueous extracts against tomato late blight in a complete randomised block design. Ten treatments were evaluated: Plasma activated water (PAW), a mixture of sodium bicarbonate and vinegar at different ratios (25/75, 50/50 and 75/25) and different mixtures of aqueous plant extracts made up of ten plants: Ageratum conyzoides, Eucalyptus saligna, Azadirachta indica, Panax quinquefolius, Callistemon viminalis, Euphorbia hirta, E. cordifolia Laggera pterodonta, Ocimum gratissimum and avocado pit powder. Plantizeb and distilled water were used as positive and negative controls, respectively. There was a significant difference (p &lt; 0.05) in the growth variables between treatments at 10 weeks after transplanting (WAT). All the treatments reduced late blight severity compared to the negative control (100 %) at 10 WAT. Late blight severity on plants treated with Plantizeb, PAW, the mixture of bicarbonate and vinegar (B+V) at 50/50 (33.24 %) and the mixture of E. hirta + E. cordifolia extract (33.33 %) was the lowest and significantly comparable (p &lt; 0.05) to Plantizeb (31.67 %). Tomato yield was 2.24 t/ha (negative control), 28.56 t/ha (Plantizeb), 27.23 t/ha (PAW), 30.32 t / ha (B+V at 50/50), and 27.69 t/ha (mixture of the aqueous extract of E. hirta + E. cordifolia). The PAW, the mixtures between B+V at 50/50, and the aqueous extract of E. hirta and E. cordifolia could be exploited for their bioactivity in late blight management in field conditions.

Ultralong-lasting plasma-activated water: production and control mechanisms

Despite the rapidly growing interest stemming from the broad-spectrum, high inactivation capacity, and environmental friendliness of the plasma-activated water (PAW), practical applications are limited because of the PAW’s short lifetime. While low-temperature storage can extend the lifetime, but the freezing and thawing processes are energy- and labor-intense and are generally not suitable for large-scale applications such as environmental and biomedical disinfection. This work addresses this issue by developing the ultra-long-life PAW at room temperature. The innovative approach is based on using DC needle-water discharges, wherein the gaseous products are blown out and absorbed separately by a gas flow. By simply adjusting the voltage and gas flow rates, two distinctive types of PAW with acidic hydrogen peroxide and nitrite as the main products are produced and separated in the discharge chamber and gas bubbling chamber. Intentional mixing of these two PAWs causes a chain chemical reaction dominated by peroxynitrite (ONOOH). This reaction can generate a variety of short-lived reactive species, thereby achieving the ultralong-lasting PAW with very stable bactericidal ability. This study further demonstrates the ability to effectively control the reaction products in both chambers and provides insights into the secondary activation mechanism of short-lived reactive species stimulated by ONOOH.