Effects and mechanisms of jujube juice components on degradation of Alternaria mycotoxin by cold plasma

Effects and mechanisms of jujube juice components on degradation of Alternaria mycotoxin by cold plasma | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effects and mechanisms of jujube juice components on degradation of Alternaria mycotoxin by cold plasma Xiaoyuan Wang, Qing Liu, Yike Han, Zhenzhen Ge, Xiaopeng Wei, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4146628/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The study examined the impact of jujube constituents on cold plasma (CP) degradation of alternariol (AOH) and alternariol monomethyl ether (AME), followed by assessing changes in CP electrochemical properties. The correlation between these properties and toxin degradation was analyzed to understand how jujube components influence the breakdown of AOH and AME. Results showed that when treated with CP for 3 minutes, 0.00350 mg/mL jujube protein degraded 49.0% of AOH and 48.8% of AME. Under the same treatment time, 0.0500 mg/mL jujube polysaccharide degraded AOH by 74.6% and AME by 95.8%. Conversely, Vc, K + , Ca 2+ , oleic acid, and linoleic acid exhibited negligible inhibitory effects on toxin degradation. Furthermore, following CP treatment, oxidation-reduction potential (△ORP), pH, conductivity, hydrogen peroxide (H 2 O 2 ), hydroxyl radical (•OH), nitrate ion (NO 3 − ), and nitrite ion (NO 2 − ) contents in the jujube protein or polysaccharide system were significantly correlated with toxin degradation; thus confirming their association with reactive oxygen-nitrogen species (RONS). The presence of jujube protein or polysaccharide hindered the degradation of CP-mediated toxins by consuming reactive RONS resources. This study provides insights into how AOH and AME in jujube juice are degraded by CP, enabling more targeted and efficient elimination of foodborne toxins. Alternaria mycotoxins alternariol (AOH) alternariol monomethyl ether (AME) cold plasma mycotoxin degradation jujube juice components Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Introduction Alternaria mycotoxin is a secondary metabolite produced by the genus Alternaria (Man et al., 2017 ). Alternariol (AOH) and alternariol monomethyl ether (AME), two prominent toxins synthesized by fungi belonging to the Alternaria genus, have been detected naturally in various harvested crops such as wheat, grapes, jujube, Fabaceae plants, and tomatoes (Wang et al., 2022 ). Extensive literature has demonstrated that AOH and AME exhibit carcinogenicity, teratogenicity, mutagenicity, genotoxicity, as well as reproductive and developmental toxicity (Wang et al., 2020 ), which have been proven to persistently contaminate food items with limited removal options once introduced (Luo et al., 2018 ). Currently employed methods for controlling Alternaria mycotoxins include UV light exposure, inactivated yeast powder treatment, ozone application among others (Han et al., 2023 ; Wang et al., 2020 ). Cold plasma (CP) is an emerging environmentally friendly agricultural technology with promising applications in the field of food safety (Okyere et al., 2022 ). CP generates various active particles, including reactive oxygen species (ROS, such as •OH, O, 1 O 2 , •O 2 − , H 2 O 2 , and O 3 ), as well as reactive nitrogen species (RNS), like NO 2 − , and NO 3 − (Feizollahi et al., 2023 ; Liao et al., 2018 ; Wang et al., 2020 ). Under these conditions, CP effectively degrades toxic substances and inactivates microorganisms, making it a valuable tool for ensuring food safety. Kis et al. ( 2020 ) demonstrated that CP using nitrogen gas efficiently degraded T-2 and HT-2 toxins in oat flour. Feizollahi and Roopesh ( 2021 ) applied atmospheric CP to degrade zearalenone and observed complete degradation rates of up to 100% or 66.8% after treatment in solution or dry conditions for 30 s, respectively. Siciliano et al. ( 2016 ) employed CP to eliminate aflatoxin from hazelnuts by approximately 70% reduction rate. Ten Bosch et al. ( 2017 ) utilized CP to degrade mycotoxins produced by Fusarium spp., including zearalenone, deoxynivalenol, fumonisin B1, and T-2 toxin, and achieved significant degradation efficiency. Previous study by this research team demonstrated the effective degradation of AOH and AME without any noticeable impact on the quality of jujube juice (Wang et al., 2020 ). However, the degradation of Alternaria mycotoxins in aqueous solution alone was significantly higher compared to that in jujube juice. Similar findings were reported by Pankaj et al. ( 2018 ), who emphasized the importance of food substrate in determining aflatoxin reduction during thermal processing. Nevertheless, it remains unclear how each component of jujube juice influences plasma degradation of these toxins, as well as the underlying mechanism behind this interaction. To gain further insights into the impact of jujube juice components on the degradation of Alternaria mycotoxins by CP and its underlying mechanism, this study selected key constituents in jujube juice to investigate their influence on both toxin degradation and electrochemical properties of each reaction system following CP treatment. These findings are expected to lay a foundation for elucidating the mechanism behind Alternaria toxin degradation in jujube juice by CP and facilitate the development of more efficient and targeted technologies for mitigating foodborne toxins. Materials and methods Chemicals and reagents The pure mycotoxin standards of AOH and AME were procured from Pribolab Pte. Ltd. (Singapore). AOH and AME were dissolved in methanol to obtain a concentration of 100 µg/mL and stored at -20°C. Dialysis bags with molecular weight cut-off (MWCO) values of 8000 and 14000 Da, H 2 O 2 Quantitative Assay Kit (Water-Compatible), and Nitrate Determination Kit were obtained from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, CHN). All other reagents were purchased from the local regent company. Extraction and purification of jujube polysaccharide The jujube polysaccharide was extracted using ultrasonically assisted extraction and ethanol precipitation method (Ji et al., 2017 ). Specifically, the extraction procedure was conducted in the water bath at 90 ℃ for 30 min with the material-liquid ratio of 25:1 (w/w), under ultrasonic treatment at 120 W for 15 min. The extraction process was repeated three times. Afterwards, the mixture was centrifuged at 6288 x g for 20 min and the supernatant was collected and concentrated. Then added absolute ethanol and centrifuged after being placed for 3 h. The precipitate was collected and evaporated, then the polysaccharide precipitation was dissolved in distilled (DI) water and freeze-dried. Afterwards, the cured jujube polysaccharide was purified with AB-8 macroporous resin, followed by concentrated and freeze-dried. The purity of polysaccharide was measured at 490 nm by a multifunctional microplate reader (Tecan Spark 20M, Tecan, Switzerland) after adding 5% phenol solution and concentrated sulfuric acid. The absorbance values were calibrated against a glucose standard curve, with results expressed as glucose equivalents. Extraction and purification of jujube protein Jujube fruits were thoroughly washed, deseeded, cut into small pieces, and subsequently dried prior to pulverization. Jujube protein was extracted through alkali extraction followed by acid precipitation. A mixture of jujube powder and deionized water was prepared at a ratio of 1:20 (w/v), and the pH value was adjusted using NaOH and HCl to reach a level of 11. The resulting sample slurry was then subjected to stirring for 50 min in a water bath maintained at 60°C. After centrifugation at 10089 x g for 15 min, the supernatant was collected. Subsequently, proteins were precipitated by adjusting the pH to 4 followed by another round of centrifugation. The obtained precipitated proteins were dissolved in deionized water and after adjusting the pH to 7, the protein solution underwent dialysis within dialysis bags placed in a cold room set at 4°C for a duration of 24 h with regular water changes every 4 h. Finally, the dialyzed protein solution was freeze-dried. The protein content of jujube extracts was measured according to the Coomassie brilliant blue method. An aliquot of 5 mL of the Coomassie brilliant blue was mixed with 1 mL of protein solution, and the absorbance of the mixture was read at 595 nm by a multifunctional microplate reader (Tecan Spark 20M, Tecan, Switzerland). The calibration curve was performed using bovine serum albumin as a standard before the curve was brought into the standard curve and the protein content of the sample was calculated. Cold plasma treatment of samples Each jujube component was dissolved in DI water to prepare different treatment systems. The solution was then transferred into a quartz dish and subjected to dielectric barrier plasma treatment, utilizing the same equipment as previously described (Wang et al., 2020 ). The dielectric barrier discharge system primarily consisted of an AC high-voltage power supply, high-voltage electrode, grounding electrode, and insulating medium. During plasma exposure, various active substances were generated by direct contact with the solution in the quartz dish. The concentrations of different components were determined based on the nutrient content found in both jujube and jujube juice (Lu et al., 2021 ). Subsequently, the samples were subjected to DBD CP (Nanjing Suman Electronics Co., Ltd., Nanjing, CHN) treatment at a voltage of 30 kV for varying durations ranging from 0 to 10 min until complete degradation was achieved. Extraction and purification of AOH and AME After the CP treatment, all the treated samples underwent toxin extraction and purification following our previously established methods (Wang et al., 2022 ). Specifically, samples were transferred into PTFE tubes and mixed with 4 mL acetonitrile (containing 0.100% formic acid) and 1 mL DI water. The mixture was vortexed for 2 min, followed by addition of anhydrous magnesium sulfate (1.50 g) and NaCl (0.500 g), which was then vortexed again. Subsequently, the mixture was allowed to stand at room temperature for 10 min before being centrifuged at a speed of 1788 x g for 10 min. Afterwards, a volume of 2 mL supernatant was evaporated to dryness using a rotary evaporator, and the remaining toxin was dissolved in 1 mL methanol. All assays were performed in triplicate, and the samples were filtered through a membrane with a pore size of 0.220 µm prior to Ultra Performance Liquid Chromatography (UPLC) analysis. Analysis of AOH and AME The levels of AOH and AME were quantified using UPLC (1290UPLC™, Beijing Agilent Technology Co., Ltd., CHN; 1290 Bin Pump: G4220A; 1290 Sampler: G4226A; 1290 Diode array detector (DAD): G4212A) as described in our previous study (Wang et al., 2022 ). The separation was performed by a ZORBAX Eclipse Plus C18 (4.6 × 250 mm I.D. 5 µm) column. The specific UPLC conditions included the use of methanol: water = 75%: 25% iso-elution for 15 min; the injection volume of 10 µL, and the flow rate of 0.800 mL/min at a column temperature of 35 ℃ with detection wavelength set at 258.0 nm. Mean recoveries of AOH and AME from quintuplicate mixed samples ranged from 86.1–108%, and 78.5–109%, with coefficients of variation being 4.89% and 5.96%, respectively, at a level of 1000 µg·kg − 1 . The LOD of this method is 15.0 µg·kg − 1 . The degradation percentage was calculated according to formula (1): Where C 0 is the initial concentration of mycotoxins (treatment time: 0 min) and C t is the concentration of mycotoxins by CP treatment at t min. Detection of oxidation-reduction potential (ORP) In order to investigate the mechanism underlying the impact of jujube constituents on mycotoxin plasma degradation, we assessed the ORP before and after plasma treatment using different jujube components. ORP is a robust solution indicator that can be employed to evaluate both oxidant concentration and its strength or activity (Zhang et al., 2017 ). The ORP of the samples before and after CP treatment was immediately measured directly by an ORP (PHSJ-6L) meter (Shanghai Yidian Scientific Instrument Co., Ltd., CHN), with 501 ORP composite electrodes. Detection of radical scavenging ability of •OH The scavenging ability of •OH was assessed using an electron paramagnetic resonance spectrometer (ESR). Specifically, a mixture of 100 µL of 1 mol/L DMPO, 5 mmol/L FeSO 4 , and samples or DI water was rapidly shaken with an oscillator for 40 s. Subsequently, 100 µL of 50 mmol/L H 2 O 2 was added for ESR detection. ESR measurements were conducted at ambient temperature using a continuous wave mode electron spin resonance spectrometer (E-Scan; Bruker, Beijing, CHN) operating at a frequency of 9.79 GHz with the Super High Q cavity employed. ESR spectra were recorded under the following settings: receiver gain set to 1×10 3 , power set to 5.00 mW, center field at 3487 G, sweep width of 100 G, sweep time lasting for approximately 10.49 s, time constant set to 20.48, modulation frequency set to 86.0 kHz, modulation amplitude adjusted to 1.01 G, and abscissa points number set as 512. The •OH radical scavenging rate was calculated according to formula (2): Detection of H 2 O 2 , NO 3 − and NO 2 − The concentration of H 2 O 2 , with and without different components of jujube juice, was determined after plasma treatment (30 kV, 2 min) using the H 2 O 2 quantitative assay kit (Water-Compatible). Similarly, the concentrations of NO 3 − and NO 2 − were measured with nitrate and nitrite assay kits respectively, both before and after plasma treatment (30 kV, 2 min) in the presence or absence of various jujube juice components. Statistical analysis All the assays in this study were repeated three times. Statistical analysis was carried out using SPSS 18.0, and a significant difference was verified by one-way ANOVA with Duncan's multiple range test ( P < 0.05). Graphs were generated using Origin Pro 2017. Results and discussion Effect of jujube polysaccharide on CP degradation of AOH and AME Polysaccharide, an essential constituent of jujube nutrients, was found to influence the degradation of AOH and AME by CP as depicted in Fig. 1 . In aqueous solutions, CP treatment at 30 kV for 4 min resulted in complete degradation of both AOH and AME. However, their degradation rates decreased upon exposure to jujube polysaccharide. While AOH was completely degraded within 7 min, AME required only 5 min, indicating a relatively lower impact of jujube polysaccharide on AME degradation compared to AOH. Nevertheless, the inhibitory effect on the degradation of both toxins did not increase with increasing concentrations of jujube polysaccharide. Based on these findings, it was evident that all tested jujube polysaccharides effectively inhibited the degradation of Alternaria mycotoxins by CP. This inhibition might be attributed to the enhanced barrier formed by jujube polysaccharide molecules, resulting in improved toxin protection. Fan et al. ( 2020 ) reported that CP treatment led to a more compact and orderly arrangement of polysaccharides, with a transformation of monosaccharide ring from β-pyran sugar to β-furan sugar. Benoit et al. ( 2011 ) demonstrated that cellulose could undergo oxidation due to reactive species generated by plasma, while •OH played a critical role in the plasma hydrolysis process of polysaccharides. From this perspective, carbohydrates might partially consume active particles produced by CP, thereby inhibiting plasma-mediated degradation of mycotoxins. The prolonged time required for complete toxin degradation could be attributed to the depletion of highly reactive particles generated by CP since jujube polysaccharide exhibits effective reducing power and various antioxidant activities such as free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating. Effect of jujube protein on the degradation of AOH and AME by CP The degradation percentages of AOH and AME by CP at different concentrations of jujube protein were illustrated in Fig. 2 , demonstrating the significant impact of jujube protein on the plasma-mediated degradation of AOH and AME. As the concentration of jujube protein increased from 0 g/mL to 0.00750 g/mL, the degradation percentages of AOH after 5 minutes of plasma treatment decreased from 100–43.2%, while AME degradation reduced from 100–43.1%. After being treated with a voltage of 30 kV for 9 minutes, AOH was degraded by 98.2%, 93.4%, and 94.2% when exposed to jujube protein concentrations of 0.00350, 0.00550, and 0.00750 g/mL respectively; whereas AME was degraded by 96.9%, 92.3%, and 87.2%, respectively. When treatment time was extended to 10 min, both toxins could be completely degraded. Based on these findings, it is evident that all tested concentrations of jujube protein could inhibit the degradation of Alternaria mycotoxins by CP to some extent. The inhibition of jujube protein on mycotoxin degradation by cold plasma may be attributed to the protection of proteins. During CP discharge, ROS including •OH, atomic oxygen (O), 1 O 2 , •O 2 − and H 2 O 2 , accompany O 3 was produced (Jiang et al., 2020 ; Misra et al., 2019 ). These ROS can disrupt the initial protein structure and enhance the mechanical properties of the interfacial film, thereby increasing their activity and expansion at interfaces (Jiang et al., 2020 ). Following CP treatment, the protein structure underwent changes leading to the formation of a robust network structure that acted as a stronger barrier between mycotoxin molecules and the surrounding environment. The interfacial film formed by CP is likely to be the primary factor inhibiting AOH and AME degradation. Moreover, •OH is presumed to attack C-H bonds during protein modification by CP, forming CH-OH H-bonded complexes followed by hydrogen motion. Additionally, H 2 O 2 is also related to the dissociation of the C-O bond, but with less effectiveness (Yusupov et al., 2013 ). It is suggested that jujube protein could scavenge free radicals produced by CP. However, as treatment time prolongs, active particles generated by plasma become sufficient to overcome this blocking effect of proteins and completely degrade toxins. Effect of Vc on the degradation of AOH and AME by CP Chinese jujube has been found to possess a high content of Vc (Chen et al., 2019 ), which may influence the degradation of mycotoxins by CP. Therefore, it is imperative to investigate the impact of Vc on the degradation of Alternaria mycotoxins in jujube juice through CP treatment. As depicted in Fig. 3 , in an aqueous solution, the degradation percentages of AOH were 73.8%, 93.3%, 96.7%, and 100% after plasma treatment for 1, 2, 3, and 4 min respectively; while the degradation rates of AME were recorded as 78.4%, 94.0%, 98.4%, and ultimately reaching complete degradation at a rate of 100%. Under identical treatment conditions with a Vc concentration of 0.350 mg/mL, the degradation percentage of AOH were observed as 0%, 72.3%, 84.3%, 91.7%, 97.6%, and 100% respectively, whereas for AME, the corresponding values were 0%, 67.5%, 80.0%, 84.5%, 96.2%, and ultimately reaching complete degradation respectively. When exposed to varying concentrations of Vc, the plasma degradation of AOH and AME increased with discharge time until complete degradation was achieved after 5 min, a slightly longer duration compared to that in aqueous solution. The results indicated that the degradation of AOH by cold plasma was not significantly impacted by Vc, while AME degradation showed slight susceptibility. The potential inhibition of AME degradation by Vc might be attributed to the antioxidant capacity of VC, wherein the free radicals generated by CP could be scavenged, resulting in a partial reduction in their attack on both AOH and AME. The disparity between AOH and AME might be ascribed to the presence of three hydroxyl groups on the benzene ring of AOH, rendering it more susceptible to oxidation compared to AME which possesses only two hydroxyl groups along with one methoxy group. Similar findings have been reported by Surowsky ( 2016 ) who suggested that Vc exhibited protective effects against reactive oxygen species-induced damage in food matrices owing to its inherent antioxidant properties. Effect of Ca 2+ and K + on the degradation of AOH and AME by CP Jujube fruit has been demonstrated to possess a high content of essential micronutrients, including Ca 2+ and K + , which play crucial roles in promoting health and preventing diseases(Lu et al., 2021 ). The degradation of AOH in aqueous solution, with or without the presence of Ca 2+ , was effectively facilitated by CP in a time-dependent manner, as demonstrated in Fig. 4 . The degradation percentages of AOH when exposed to Ca + concentrations of Ca 2+ of 0.0050, 0.00750, and 0.100 mg/mL were found to be 73.8%, 73.4%, 80.5% and 77.6%, respectively, whereas for AME, the corresponding degradation percentages were observed as 78.4%, 62.3%, 58.7%, and 60.7%, respectively. After being treated by CP at 30 kV for 4 min, the degradation percentages of AME in 0, 0.100, 0.250, and 0.400 mg/mL KCl solution were 100%, 96.4%, 97.8%, and 97.6%, respectively, suggesting that KCl slightly decreased AME degradation. For both Ca 2+ and K + , no obvious effect was observed on AOH degradation, while slight inhibition on AME degradation was presented. It has been reported that the degradation of mycotoxin by CP is attributed to the oxidation ability of ROS and RNS generated by CP. Neves et al. ( 2022 ) have suggested that Ca 2+ may mitigate the toxic levels of NO 2 − in pacu juveniles, potentially through alleviating oxidative stress-induced ROS or RNS production. This implies that Ca 2+ could reduce the levels of RON and ROS, leading to a slight decrease in AME degradation rate. Feng et al. ( 2006 ) have mentioned that KCl acts as a scavenger for •OH. However, it has been established that •OH produced by CP plays a major role in mycotoxin degradation (Attri et al., 2015 ). It was hypothesized that the partial consumption of •OH by KCl may be the reason for the reduction of Alternaria mycotoxin degradation. Moreover, the impact of K + on AME degradation surpasses AOH, which could be attributed to the presence of three hydroxyl groups on AOH's benzene ring compared to two hydroxyl groups plus one methoxy group on AME. Effect of oleic acid and linoleic acid on the degradation of AOH and AME by CP The impact of oleic acid on the plasma degradation of AOH and AME was illustrated in Fig. 5 . Even when exposed to varying concentrations of oleic acid, AOH could be completely degraded after a 3-min treatment with CP. For AME, the degradation percentage was 95.5%, 93.7%, and 96.9% at oleic acid concentrations of 0.0500, 0.100, and 0.150 mg/mL respectively; however, complete degradation occurred within 5 min regardless of concentration level. These results demonstrated that oleic acid had little effect on the degradation of AOH and AME by CP. Figure 5 also depicted the effect of linoleic acid on plasma degradation of AOH and AME; no significant impact was observed for AOH while slight effects were noted for AME at the highest concentration tested (0.15 mg/mL) under these experimental conditions. Batista et al. ( 2021 ) demonstrated that the fatty acid profile remains unaffected by CP technology. Perez-Andres et al. ( 2020 ) investigated the impact of cold atmospheric plasma treatment on commercially packaged mackerel fillets and reported insignificant alterations in both fatty acid composition and nutritional quality indices post-treatment. The fact that the degradation of mycotoxin by CP was not significantly affected in the presence of oleic acid and linoleic acid might be because oleic acid and linoleic acid did not consume the active substances produced by CP. Mechanism of the effect of jujube components on mycotoxin plasma degradation The effect of jujube components on the electrochemical properties of the systems after cold plasma discharge ORP is a measure of a solution's ability to oxidize or reduce another substance, reflecting the overall level of RONS present in the solution. A higher ORP value indicates a stronger oxidation capacity (Thirumdas et al., 2018 ). As shown in Fig. 6 , plasma treatment resulted in varying degrees of increase in the ORP of the solution. The results of this experimental study demonstrated that the changes in △ORP for jujube protein solution, jujube polysaccharide, and simulated jujube juice were lower compared to an aqueous solution. However, solutions with different concentrations of Vc, K + , Ca 2+ , oleic acid, and linoleic acid exhibited similar changes in △ORP as observed for an aqueous solution. The variation trend of △ORP was highly consistent with the degradation pattern of toxins, suggesting that the effect of jujube juice components on AOH and AME degradation might be attributed to the oxidative capability of certain RONS generated by CP discharge. H 2 O 2 , a major active substance generated during CP discharge, exhibits strong oxidation effects (Ma et al., 2021 ). As depicted in Fig. 7 , the H 2 O 2 content of solutions containing various jujube components such as jujube protein, jujube polysaccharide, Ca 2+ , K + , Vc, oleic acid, and linoleic acid was significantly lower than that of aqueous solution after CP treatment. Tripathi and Mishra ( 2009 ) suggested that H 2 O 2 can degrade mycotoxins via oxidation. Additionally, Ma et al. ( 2021 ) highlighted the crucial role of H 2 O 2 in chitosan degradation during CP discharge and its decomposition to produce •OH. The impact of different jujube juice components on AOH and AME degradation by CP might be attributed to their partial consumption of H 2 O 2 . The effect of CP on the •OH scavenging rate of various jujube juice component model systems was shown in Fig. 7 . The results showed that with the increase of jujube protein concentration from 0 g/mL to 0.00750 g/mL, the •OH clearance decreased significantly from 70.3–24.1%. Furthermore, an increase in jujube polysaccharide concentration to 0.0750 g/mL resulted in a reduction of •OH clearance decreased by 6.19%. Conversely, no notable change was observed in •OH clearance following CP treatment across different concentrations of linoleic acid, Ca 2+ , K + , and Vc environments. As for oleic acid, there was an initial significant decrease followed by an increase in •OH clearance with increasing concentration. Notably, compared to the aqueous solution, the multi-component model system of jujube juice exhibited a substantial decrease in •OH clearance with increasing concentration. It is worth mentioning that •OH is one of the primary short-lived reactive species generated during CP discharge (Kurake et al., 2017 ). Several scholars have argued that the oxidizing effectors produced by CP are primarily attributed to •OH radicals which can disrupt structurally important bonds within peptidoglycan (i.e., C-O, C-N, or C-C bonds) (Yusupov et al., 2013 ). Moreover, variations observed in •OH clearance were highly consistent with the degradation effects exerted by AOH and AME under different jujube component model systems; thus indicating a strong correlation between their removal from jujube juice through CP treatment and •OH generation during discharge. In addition to ROS, CP discharge also generates extremely rich RNS (NO 3 − and NO 2 − , etc.) (Sardella et al., 2021 ; Yusupov et al., 2013 ; Lukes et al., 2014 ). Therefore, it is imperative to investigate the impact of CP on NO 3 − and NO 2 − in various model systems representing jujube juice components (Fig. 8 ). The results revealed that following CP treatment, the concentration of NO 2 − significantly decreased for different jujube juice components compared to the aqueous solution. This reduction could be attributed to two factors. Firstly, diverse jujube juice components might consume some NO 2 − through chemical reactions with them. Secondly, there might be a conversion of NO 2 − into NO 3 − . Luke et al. (2014) proposed that in aqueous solutions, unstable NO 2 − could undergo transformation into more stable form i.e., NO 3 − , leading to pH decrease and increased acidity. The impact of CP on the NO 3 − content in various jujube juice component model systems was illustrated in Fig. 8 . Following an equivalent duration of CP treatment, distinct jujube juice component solutions exhibited varying levels of NO 3 − content, with significantly lower concentrations observed when exposed to jujube polysaccharide and protein compared to aqueous solution. However, the presence of Ca 2+ , K + , oleic acid, linoleic acid, and Vc had minimal influence on the NO 3 − concentration generated by CP. Consequently, the degradation of AOH and AME by CP in jujube juice was found to be closely associated with the production of NO 3 − during CP discharge. The correlation analysis between the changes of electrochemical properties of the system and the CP degradation of toxins After CP treatment, the Pearson correlation analysis revealed in Fig. 9 and Fig. 10 demonstrated that the degradation of AOH and AME was significantly positively correlated with H 2 O 2 , •OH, NO 3 − , NO 2 −, and ORP while being negatively correlated with pH when exposed to jujube protein and polysaccharide. Furthermore, a significant correlation was observed between the physical and chemical properties as well as active substance content of jujube protein and polysaccharide solution after CP treatment. This suggested that the inhibition of toxin degradation by jujube protein and polysaccharide was closely related to changes in physicochemical properties and active substances produced by CP. In the Pearson correlation analysis of AOH and AME degradation and the changes in Vc model system properties, the degradation trend of AOH exhibited significant positive correlations with H 2 O 2 , •OH, NO 3 − , and NO 2 − contents, while showing a negative correlation with pH value. However, no significant correlations were observed between △ORP and conductivity. Meanwhile, the degradation trend of AME showed positive correlations with H 2 O 2 content, NO 2 − content, and △ORP; whereas it displayed a negative correlation with pH. Regarding the Ca 2+ , K + , oleic acid, and linoleic acid model systems, the degradation trend of AOH showed only partial or insignificant correlation with certain properties or active substances. In summary, following CP treatment, there was a significant positive correlation between the change in △ORP, the contents of H 2 O 2 , •OH, NO 3 − , and NO 2 − of jujube polysaccharide, jujube protein, and multi-component model system and the degradation trend of toxins. Additionally, the degradation trend of toxins in Vc system exhibited a high correlation with H 2 O 2 , •OH, NO 3 − , and NO 2 − . However for oleic acid, linoleic acid, Ca 2+ , and K + , the degradation trend of toxins displayed less correlation with the properties of these systems. Furthermore, the results from Pearson correlation analysis further demonstrated that jujube components could influence the degradation of AOH and AME by CP through consumption of active particles including H 2 O 2 , •OH, NO 3 − , and NO 2 − . It can be inferred that RONS produced by CP is considered as a primary factor responsible for toxin degradation. Overall, the present study demonstrated the effective degradation of AOH and AME in both aqueous solution and jujube juice by CP. However, the degradation was partially inhibited in jujube juice. Among these tested components of jujube juice, jujube protein exhibited the highest inhibitory effect on plasma degradation of these two toxins, followed by jujube polysaccharide. Other components including Vc, Ca 2+ , K + , oleic acid, or linoleic acid showed insignificant effects on AOH and AME degradation. Furthermore, the degradation of AOH and AME by CP was closely associated with the presence of H 2 O 2 , •OH, NO 3 − , and NO 2 − active particles in different discharge modes during CP treatment. After CP treatment, consumption of RONS such as H 2 O 2 , •OH, NO 3 − , and NO 2 − by jujube protein or polysaccharide may be a key factor contributing to inhibition of AOH and AME degradation. This study provides a theoretical basis for understanding the mechanism underlying Alternaria mycotoxin degradation in jujube juice system using CP technology and promotes its application for efficient targeted detoxification in food systems. Declarations Acknowledgements We appreciate the help of Key Scientific Research Projects of Colleges and Universities in Henan province (24A550020) established by Department of Education of Henan Province. CRediT authorship contribution statement X. Y. W. : Investigation, methodology, writing, review & editing, funding acquisition, resources. Q. L. : methodology, formal analysis. Y. K. H. : Investigation, methodology, formal analysis. Z. Z. G. : review & editing. X. P. W. : Writing, review & editing. Y. Z. Y. : methodology, formal analysis. W. Z. : Supervision, review & editing. Funding This research was supported by Key Scientific Research Projects of Colleges and Universities in Henan province (24A550020) established by Department of Education of Henan Province. Competing Interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. References Attri, P., Kim, Y.H., Park, D.H., Park, J.H., Hong, Y.J., Uhm, H.S., Kim, K.-N., Fridman, A. and Choi, E.H., 2015. Generation mechanism of hydroxyl radical species and its lifetime prediction during the plasma-initiated ultraviolet (UV) photolysis. Scientific Reports 5: 9332. https://doi.org/10.1038/srep09332 Batista, J.D.F., Dantas, A.M., Fonseca, J.V.D., Madruga, M.S., Fernandes, F.A.N., Rodrigues, S. and Borges, G.D., 2021. Effects of cold plasma on avocado pulp (Persea americana Mill.): chemical characteristics and bioactive compounds. 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Current Research in Food Science 5: 451-463. https://doi.org/10.1016/j.crfs.2022.02.007 Pankaj, S.K., Shi, H. and Keener, K.M., 2018. A review of novel physical and chemical decontamination technologies for aflatoxin in food. Trends in Food Science and Technology 71: 73-83. https://doi.org/10.1016/j.tifs.2017.11.007 Perez-Andres, J.M., de Alba, M., Harrison, S.M., Brunton, N.P., Cullen, P.J. and Tiwari, B.K., 2020. Effects of cold atmospheric plasma on mackerel lipid and protein oxidation during storage. Lwt-Food Science and Technology 118. https://doi.org/10.1016/j.lwt.2019.108697 Sardella, E., Veronico, V., Gristina, R., Grossi, L., Cosmai, S., Striccoli, M., Buttiglione, M., Fracassi, F. and Favia, P., 2021. Plasma Treated Water Solutions in Cancer Treatments: The Contrasting Role of RNS. Antioxidants 10. https://doi.org/10.3390/antiox10040605 Siciliano, I., Spadaro, D., Prelle, A., Vallauri, D., Cavallero, M.C., Garibaldi, A. and Gullino, M.L., 2016. Use of cold atmospheric plasma to detoxify hazelnuts from aflatoxins. Toxins 8: 125. https://doi.org/10.3390/toxins8050125 Surowsky, B., 2016. Chapter 7–Cold plasma interactions with food constituents in liquid and solid food matrices. Cold Plasma in Food & Agriculture 7: 179-203. https://doi.org/10.1016/B978-0-12-801365-6.00007-X Ten Bosch, L., Pfohl, K., Avramidis, G., Wieneke, S., Vioel, W. and Karlovsky, P., 2017. Plasma-Based Degradation of Mycotoxins Produced by Fusarium , Aspergillus and Alternaria Species. Toxins 9: 97. https://doi.org/10.3390/toxins9030097 Thirumdas, R., Kothakota, A., Annapure, U., Siliveru, K., Blundell, R., Gatt, R. and Valdramidis, V.P., 2018. Plasma activated water (PAW): chemistry, physico-chemical properties, applications in food and agriculture. Trends in Food Science and Technology 77: 21-31. https://doi.org/10.1016/j.tifs.2018.05.007 Tripathi, S. and Mishra, H.N., 2009. Studies on the efficacy of physical, chemical and biological aflatoxin B1 detoxification approaches in red chilli powder. International Journal of Food Safety Nutrition & Public Health 2: 325-327. https://doi.org/10.1504/IJFSNPH.2009.026920 Wang, X.Y., Han, Y.K., Niu, H., Zhang, L.H., Xiang, Q.S. and Zong, W., 2022. Alternaria mycotoxin degradation and quality evaluation of jujube juice by cold plasma treatment. Food Control 137: 108926. https://doi.org/10.1016/j.foodcont.2022.108926 Wang, X.Y., Wang, S.H., Yan, Y.Z., Wang, W.J., Zhang, L.H. and Zong, W., 2020. The degradation of Alternaria mycotoxins by dielectric barrier discharge cold plasma. Food Control 117: 107333. https://doi.org/10.1016/j.foodcont.2020.107333 Yusupov, M., Bogaerts, A., Huygh, S., Snoeckx, R., van Duin, A.C.T. and Neyts, E.C., 2013. Plasma-induced destruction of bacterial cell wall components: a reactive molecular dynamics simulation. Journal of Physical Chemistry 117: 5993-5998. https://doi.org/10.1021/jp3128516 Zhang, Y.C., Li, Y., Li, Y.L., Yu, S., Li, H.Y. and Zhang, J., 2017. A novel approach to the pacemaker infection with non-thermal atmospheric pressure plasma. European Physical Journal Special Topics 226: 2901-2910. https://doi.org/10.1140/epjst/e2016-60331-4 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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class=\"CitationRef\"\u003e2017\u003c/span\u003e). Alternariol (AOH) and alternariol monomethyl ether (AME), two prominent toxins synthesized by fungi belonging to the \u003cem\u003eAlternaria\u003c/em\u003e genus, have been detected naturally in various harvested crops such as wheat, grapes, jujube, Fabaceae plants, and tomatoes (Wang et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Extensive literature has demonstrated that AOH and AME exhibit carcinogenicity, teratogenicity, mutagenicity, genotoxicity, as well as reproductive and developmental toxicity (Wang et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), which have been proven to persistently contaminate food items with limited removal options once introduced (Luo et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Currently employed methods for controlling \u003cem\u003eAlternaria\u003c/em\u003e mycotoxins include UV light exposure, inactivated yeast powder treatment, ozone application among others (Han et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCold plasma (CP) is an emerging environmentally friendly agricultural technology with promising applications in the field of food safety (Okyere et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). CP generates various active particles, including reactive oxygen species (ROS, such as \u0026bull;OH, O, \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;O\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, and O\u003csub\u003e3\u003c/sub\u003e), as well as reactive nitrogen species (RNS), like NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e (Feizollahi et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Liao et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Under these conditions, CP effectively degrades toxic substances and inactivates microorganisms, making it a valuable tool for ensuring food safety. Kis et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) demonstrated that CP using nitrogen gas efficiently degraded T-2 and HT-2 toxins in oat flour. Feizollahi and Roopesh (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) applied atmospheric CP to degrade zearalenone and observed complete degradation rates of up to 100% or 66.8% after treatment in solution or dry conditions for 30 s, respectively. Siciliano et al. (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) employed CP to eliminate aflatoxin from hazelnuts by approximately 70% reduction rate. Ten Bosch et al. (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) utilized CP to degrade mycotoxins produced by \u003cem\u003eFusarium\u003c/em\u003e spp., including zearalenone, deoxynivalenol, fumonisin B1, and T-2 toxin, and achieved significant degradation efficiency.\u003c/p\u003e \u003cp\u003ePrevious study by this research team demonstrated the effective degradation of AOH and AME without any noticeable impact on the quality of jujube juice (Wang et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, the degradation of \u003cem\u003eAlternaria\u003c/em\u003e mycotoxins in aqueous solution alone was significantly higher compared to that in jujube juice. Similar findings were reported by Pankaj et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), who emphasized the importance of food substrate in determining aflatoxin reduction during thermal processing. Nevertheless, it remains unclear how each component of jujube juice influences plasma degradation of these toxins, as well as the underlying mechanism behind this interaction.\u003c/p\u003e \u003cp\u003eTo gain further insights into the impact of jujube juice components on the degradation of \u003cem\u003eAlternaria\u003c/em\u003e mycotoxins by CP and its underlying mechanism, this study selected key constituents in jujube juice to investigate their influence on both toxin degradation and electrochemical properties of each reaction system following CP treatment. These findings are expected to lay a foundation for elucidating the mechanism behind \u003cem\u003eAlternaria\u003c/em\u003e toxin degradation in jujube juice by CP and facilitate the development of more efficient and targeted technologies for mitigating foodborne toxins.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003eChemicals and reagents\u003c/h2\u003e\n \u003cp\u003eThe pure mycotoxin standards of AOH and AME were procured from Pribolab Pte. Ltd. (Singapore). AOH and AME were dissolved in methanol to obtain a concentration of 100 \u0026micro;g/mL and stored at -20\u0026deg;C. Dialysis bags with molecular weight cut-off (MWCO) values of 8000 and 14000 Da, H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e Quantitative Assay Kit (Water-Compatible), and Nitrate Determination Kit were obtained from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, CHN). All other reagents were purchased from the local regent company.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003eExtraction and purification of jujube polysaccharide\u003c/h2\u003e\n \u003cp\u003eThe jujube polysaccharide was extracted using ultrasonically assisted extraction and ethanol precipitation method (Ji et al., \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e). Specifically, the extraction procedure was conducted in the water bath at 90 ℃ for 30 min with the material-liquid ratio of 25:1 (w/w), under ultrasonic treatment at 120 W for 15 min. The extraction process was repeated three times. Afterwards, the mixture was centrifuged at 6288 x g for 20 min and the supernatant was collected and concentrated. Then added absolute ethanol and centrifuged after being placed for 3 h. The precipitate was collected and evaporated, then the polysaccharide precipitation was dissolved in distilled (DI) water and freeze-dried. Afterwards, the cured jujube polysaccharide was purified with AB-8 macroporous resin, followed by concentrated and freeze-dried. The purity of polysaccharide was measured at 490 nm by a multifunctional microplate reader (Tecan Spark 20M, Tecan, Switzerland) after adding 5% phenol solution and concentrated sulfuric acid. The absorbance values were calibrated against a glucose standard curve, with results expressed as glucose equivalents.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003eExtraction and purification of jujube protein\u003c/h2\u003e\n \u003cp\u003eJujube fruits were thoroughly washed, deseeded, cut into small pieces, and subsequently dried prior to pulverization. Jujube protein was extracted through alkali extraction followed by acid precipitation. A mixture of jujube powder and deionized water was prepared at a ratio of 1:20 (w/v), and the pH value was adjusted using NaOH and HCl to reach a level of 11. The resulting sample slurry was then subjected to stirring for 50 min in a water bath maintained at 60\u0026deg;C. After centrifugation at 10089 x g for 15 min, the supernatant was collected. Subsequently, proteins were precipitated by adjusting the pH to 4 followed by another round of centrifugation. The obtained precipitated proteins were dissolved in deionized water and after adjusting the pH to 7, the protein solution underwent dialysis within dialysis bags placed in a cold room set at 4\u0026deg;C for a duration of 24 h with regular water changes every 4 h. Finally, the dialyzed protein solution was freeze-dried.\u003c/p\u003e\n \u003cp\u003eThe protein content of jujube extracts was measured according to the Coomassie brilliant blue method. An aliquot of 5 mL of the Coomassie brilliant blue was mixed with 1 mL of protein solution, and the absorbance of the mixture was read at 595 nm by a multifunctional microplate reader (Tecan Spark 20M, Tecan, Switzerland). The calibration curve was performed using bovine serum albumin as a standard before the curve was brought into the standard curve and the protein content of the sample was calculated.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003eCold plasma treatment of samples\u003c/h2\u003e\n \u003cp\u003eEach jujube component was dissolved in DI water to prepare different treatment systems. The solution was then transferred into a quartz dish and subjected to dielectric barrier plasma treatment, utilizing the same equipment as previously described (Wang et al., \u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). The dielectric barrier discharge system primarily consisted of an AC high-voltage power supply, high-voltage electrode, grounding electrode, and insulating medium. During plasma exposure, various active substances were generated by direct contact with the solution in the quartz dish. The concentrations of different components were determined based on the nutrient content found in both jujube and jujube juice (Lu et al., \u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e). Subsequently, the samples were subjected to DBD CP (Nanjing Suman Electronics Co., Ltd., Nanjing, CHN) treatment at a voltage of 30 kV for varying durations ranging from 0 to 10 min until complete degradation was achieved.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003eExtraction and purification of AOH and AME\u003c/h2\u003e\n \u003cp\u003eAfter the CP treatment, all the treated samples underwent toxin extraction and purification following our previously established methods (Wang et al., \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e). Specifically, samples were transferred into PTFE tubes and mixed with 4 mL acetonitrile (containing 0.100% formic acid) and 1 mL DI water. The mixture was vortexed for 2 min, followed by addition of anhydrous magnesium sulfate (1.50 g) and NaCl (0.500 g), which was then vortexed again. Subsequently, the mixture was allowed to stand at room temperature for 10 min before being centrifuged at a speed of 1788 x g for 10 min. Afterwards, a volume of 2 mL supernatant was evaporated to dryness using a rotary evaporator, and the remaining toxin was dissolved in 1 mL methanol. All assays were performed in triplicate, and the samples were filtered through a membrane with a pore size of 0.220 \u0026micro;m prior to Ultra Performance Liquid Chromatography (UPLC) analysis.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003eAnalysis of AOH and AME\u003c/h2\u003e\n \u003cp\u003eThe levels of AOH and AME were quantified using UPLC (1290UPLC\u0026trade;, Beijing Agilent Technology Co., Ltd., CHN; 1290 Bin Pump: G4220A; 1290 Sampler: G4226A; 1290 Diode array detector (DAD): G4212A) as described in our previous study (Wang et al., \u003cspan class=\"CitationRef\"\u003e2022\u003c/span\u003e). The separation was performed by a ZORBAX Eclipse Plus C18 (4.6 \u0026times; 250 mm I.D. 5 \u0026micro;m) column. The specific UPLC conditions included the use of methanol: water\u0026thinsp;=\u0026thinsp;75%: 25% iso-elution for 15 min; the injection volume of 10 \u0026micro;L, and the flow rate of 0.800 mL/min at a column temperature of 35 ℃ with detection wavelength set at 258.0 nm. Mean recoveries of AOH and AME from quintuplicate mixed samples ranged from 86.1\u0026ndash;108%, and 78.5\u0026ndash;109%, with coefficients of variation being 4.89% and 5.96%, respectively, at a level of 1000 \u0026micro;g\u0026middot;kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The LOD of this method is 15.0 \u0026micro;g\u0026middot;kg\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The degradation percentage was calculated according to formula (1):\u003c/p\u003e\n \u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\n \u003cdiv class=\"EquationNumber\"\u003e\u003cimg src=\"https://myfiles.space/user_files/122228_c8a1650c59388082/122228_custom_files/img1711616890.png\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003c/div\u003e\n \u003cp\u003eWhere \u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e0\u003c/em\u003e\u003c/sub\u003e is the initial concentration of mycotoxins (treatment time: 0 min) and \u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003et\u003c/em\u003e\u003c/sub\u003e is the concentration of mycotoxins by CP treatment at t min.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n \u003ch2\u003eDetection of oxidation-reduction potential (ORP)\u003c/h2\u003e\n \u003cp\u003eIn order to investigate the mechanism underlying the impact of jujube constituents on mycotoxin plasma degradation, we assessed the ORP before and after plasma treatment using different jujube components. ORP is a robust solution indicator that can be employed to evaluate both oxidant concentration and its strength or activity (Zhang et al., \u003cspan class=\"CitationRef\"\u003e2017\u003c/span\u003e). The ORP of the samples before and after CP treatment was immediately measured directly by an ORP (PHSJ-6L) meter (Shanghai Yidian Scientific Instrument Co., Ltd., CHN), with 501 ORP composite electrodes.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003eDetection of radical scavenging ability of \u0026bull;OH\u003c/h2\u003e\n \u003cp\u003eThe scavenging ability of \u0026bull;OH was assessed using an electron paramagnetic resonance spectrometer (ESR). Specifically, a mixture of 100 \u0026micro;L of 1 mol/L DMPO, 5 mmol/L FeSO\u003csub\u003e4\u003c/sub\u003e, and samples or DI water was rapidly shaken with an oscillator for 40 s. Subsequently, 100 \u0026micro;L of 50 mmol/L H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e was added for ESR detection. ESR measurements were conducted at ambient temperature using a continuous wave mode electron spin resonance spectrometer (E-Scan; Bruker, Beijing, CHN) operating at a frequency of 9.79 GHz with the Super High Q cavity employed. ESR spectra were recorded under the following settings: receiver gain set to 1\u0026times;10\u003csup\u003e3\u003c/sup\u003e, power set to 5.00 mW, center field at 3487 G, sweep width of 100 G, sweep time lasting for approximately 10.49 s, time constant set to 20.48, modulation frequency set to 86.0 kHz, modulation amplitude adjusted to 1.01 G, and abscissa points number set as 512.\u003c/p\u003e\n \u003cp\u003eThe \u0026bull;OH radical scavenging rate was calculated according to formula (2):\u003c/p\u003e\n \u003cdiv id=\"Equ2\" class=\"Equation\"\u003e\n \u003cdiv class=\"EquationNumber\"\u003e\u003cimg src=\"https://myfiles.space/user_files/122228_c8a1650c59388082/122228_custom_files/img1711616909.png\"\u003e\u003cbr\u003e\u003c/div\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003eDetection of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e\u003c/h2\u003e\n \u003cp\u003eThe concentration of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, with and without different components of jujube juice, was determined after plasma treatment (30 kV, 2 min) using the H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e quantitative assay kit (Water-Compatible). Similarly, the concentrations of NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e were measured with nitrate and nitrite assay kits respectively, both before and after plasma treatment (30 kV, 2 min) in the presence or absence of various jujube juice components.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eStatistical analysis\u003c/h2\u003e\n \u003cp\u003eAll the assays in this study were repeated three times. Statistical analysis was carried out using SPSS 18.0, and a significant difference was verified by one-way ANOVA with Duncan\u0026apos;s multiple range test (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Graphs were generated using Origin Pro 2017.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results and discussion","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eEffect of jujube polysaccharide on CP degradation of AOH and AME\u003c/h2\u003e \u003cp\u003ePolysaccharide, an essential constituent of jujube nutrients, was found to influence the degradation of AOH and AME by CP as depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e1\u003c/span\u003e. In aqueous solutions, CP treatment at 30 kV for 4 min resulted in complete degradation of both AOH and AME. However, their degradation rates decreased upon exposure to jujube polysaccharide. While AOH was completely degraded within 7 min, AME required only 5 min, indicating a relatively lower impact of jujube polysaccharide on AME degradation compared to AOH. Nevertheless, the inhibitory effect on the degradation of both toxins did not increase with increasing concentrations of jujube polysaccharide.\u003c/p\u003e \u003cp\u003eBased on these findings, it was evident that all tested jujube polysaccharides effectively inhibited the degradation of \u003cem\u003eAlternaria\u003c/em\u003e mycotoxins by CP. This inhibition might be attributed to the enhanced barrier formed by jujube polysaccharide molecules, resulting in improved toxin protection. Fan et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) reported that CP treatment led to a more compact and orderly arrangement of polysaccharides, with a transformation of monosaccharide ring from β-pyran sugar to β-furan sugar. Benoit et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) demonstrated that cellulose could undergo oxidation due to reactive species generated by plasma, while \u0026bull;OH played a critical role in the plasma hydrolysis process of polysaccharides. From this perspective, carbohydrates might partially consume active particles produced by CP, thereby inhibiting plasma-mediated degradation of mycotoxins. The prolonged time required for complete toxin degradation could be attributed to the depletion of highly reactive particles generated by CP since jujube polysaccharide exhibits effective reducing power and various antioxidant activities such as free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eEffect of jujube protein on the degradation of AOH and AME by CP\u003c/h2\u003e \u003cp\u003eThe degradation percentages of AOH and AME by CP at different concentrations of jujube protein were illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig12\" class=\"InternalRef\"\u003e2\u003c/span\u003e, demonstrating the significant impact of jujube protein on the plasma-mediated degradation of AOH and AME. As the concentration of jujube protein increased from 0 g/mL to 0.00750 g/mL, the degradation percentages of AOH after 5 minutes of plasma treatment decreased from 100\u0026ndash;43.2%, while AME degradation reduced from 100\u0026ndash;43.1%. After being treated with a voltage of 30 kV for 9 minutes, AOH was degraded by 98.2%, 93.4%, and 94.2% when exposed to jujube protein concentrations of 0.00350, 0.00550, and 0.00750 g/mL respectively; whereas AME was degraded by 96.9%, 92.3%, and 87.2%, respectively. When treatment time was extended to 10 min, both toxins could be completely degraded. Based on these findings, it is evident that all tested concentrations of jujube protein could inhibit the degradation of \u003cem\u003eAlternaria\u003c/em\u003e mycotoxins by CP to some extent.\u003c/p\u003e \u003cp\u003eThe inhibition of jujube protein on mycotoxin degradation by cold plasma may be attributed to the protection of proteins. During CP discharge, ROS including \u0026bull;OH, atomic oxygen (O), \u003csup\u003e1\u003c/sup\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;O\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, accompany O\u003csub\u003e3\u003c/sub\u003e was produced (Jiang et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Misra et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). These ROS can disrupt the initial protein structure and enhance the mechanical properties of the interfacial film, thereby increasing their activity and expansion at interfaces (Jiang et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Following CP treatment, the protein structure underwent changes leading to the formation of a robust network structure that acted as a stronger barrier between mycotoxin molecules and the surrounding environment. The interfacial film formed by CP is likely to be the primary factor inhibiting AOH and AME degradation. Moreover, \u0026bull;OH is presumed to attack C-H bonds during protein modification by CP, forming CH-OH H-bonded complexes followed by hydrogen motion. Additionally, H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e is also related to the dissociation of the C-O bond, but with less effectiveness (Yusupov et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). It is suggested that jujube protein could scavenge free radicals produced by CP. However, as treatment time prolongs, active particles generated by plasma become sufficient to overcome this blocking effect of proteins and completely degrade toxins.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eEffect of Vc on the degradation of AOH and AME by CP\u003c/h2\u003e \u003cp\u003eChinese jujube has been found to possess a high content of Vc (Chen et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), which may influence the degradation of mycotoxins by CP. Therefore, it is imperative to investigate the impact of Vc on the degradation of \u003cem\u003eAlternaria\u003c/em\u003e mycotoxins in jujube juice through CP treatment. As depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig13\" class=\"InternalRef\"\u003e3\u003c/span\u003e, in an aqueous solution, the degradation percentages of AOH were 73.8%, 93.3%, 96.7%, and 100% after plasma treatment for 1, 2, 3, and 4 min respectively; while the degradation rates of AME were recorded as 78.4%, 94.0%, 98.4%, and ultimately reaching complete degradation at a rate of 100%. Under identical treatment conditions with a Vc concentration of 0.350 mg/mL, the degradation percentage of AOH were observed as 0%, 72.3%, 84.3%, 91.7%, 97.6%, and 100% respectively, whereas for AME, the corresponding values were 0%, 67.5%, 80.0%, 84.5%, 96.2%, and ultimately reaching complete degradation respectively. When exposed to varying concentrations of Vc, the plasma degradation of AOH and AME increased with discharge time until complete degradation was achieved after 5 min, a slightly longer duration compared to that in aqueous solution.\u003c/p\u003e \u003cp\u003eThe results indicated that the degradation of AOH by cold plasma was not significantly impacted by Vc, while AME degradation showed slight susceptibility. The potential inhibition of AME degradation by Vc might be attributed to the antioxidant capacity of VC, wherein the free radicals generated by CP could be scavenged, resulting in a partial reduction in their attack on both AOH and AME. The disparity between AOH and AME might be ascribed to the presence of three hydroxyl groups on the benzene ring of AOH, rendering it more susceptible to oxidation compared to AME which possesses only two hydroxyl groups along with one methoxy group. Similar findings have been reported by Surowsky (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) who suggested that Vc exhibited protective effects against reactive oxygen species-induced damage in food matrices owing to its inherent antioxidant properties.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eEffect of Ca\u003csup\u003e2+\u003c/sup\u003e and K\u003csup\u003e+\u003c/sup\u003e on the degradation of AOH and AME by CP\u003c/h2\u003e \u003cp\u003eJujube fruit has been demonstrated to possess a high content of essential micronutrients, including Ca\u003csup\u003e2+\u003c/sup\u003e and K\u003csup\u003e+\u003c/sup\u003e, which play crucial roles in promoting health and preventing diseases(Lu et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The degradation of AOH in aqueous solution, with or without the presence of Ca\u003csup\u003e2+\u003c/sup\u003e, was effectively facilitated by CP in a time-dependent manner, as demonstrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig14\" class=\"InternalRef\"\u003e4\u003c/span\u003e. The degradation percentages of AOH when exposed to Ca\u0026thinsp;+\u0026thinsp;concentrations of Ca\u003csup\u003e2+\u003c/sup\u003e of 0.0050, 0.00750, and 0.100 mg/mL were found to be 73.8%, 73.4%, 80.5% and 77.6%, respectively, whereas for AME, the corresponding degradation percentages were observed as 78.4%, 62.3%, 58.7%, and 60.7%, respectively. After being treated by CP at 30 kV for 4 min, the degradation percentages of AME in 0, 0.100, 0.250, and 0.400 mg/mL KCl solution were 100%, 96.4%, 97.8%, and 97.6%, respectively, suggesting that KCl slightly decreased AME degradation. For both Ca\u003csup\u003e2+\u003c/sup\u003e and K\u003csup\u003e+\u003c/sup\u003e, no obvious effect was observed on AOH degradation, while slight inhibition on AME degradation was presented.\u003c/p\u003e \u003cp\u003eIt has been reported that the degradation of mycotoxin by CP is attributed to the oxidation ability of ROS and RNS generated by CP. Neves et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) have suggested that Ca\u003csup\u003e2+\u003c/sup\u003e may mitigate the toxic levels of NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in pacu juveniles, potentially through alleviating oxidative stress-induced ROS or RNS production. This implies that Ca\u003csup\u003e2+\u003c/sup\u003e could reduce the levels of RON and ROS, leading to a slight decrease in AME degradation rate. Feng et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) have mentioned that KCl acts as a scavenger for \u0026bull;OH. However, it has been established that \u0026bull;OH produced by CP plays a major role in mycotoxin degradation (Attri et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). It was hypothesized that the partial consumption of \u0026bull;OH by KCl may be the reason for the reduction of \u003cem\u003eAlternaria\u003c/em\u003e mycotoxin degradation. Moreover, the impact of K\u003csup\u003e+\u003c/sup\u003e on AME degradation surpasses AOH, which could be attributed to the presence of three hydroxyl groups on AOH's benzene ring compared to two hydroxyl groups plus one methoxy group on AME.\u003c/p\u003e \u003cp\u003e \u003cb\u003eEffect of oleic acid and linoleic acid on the degradation of AOH and AME by CP\u003c/b\u003e \u003c/p\u003e \u003cp\u003eThe impact of oleic acid on the plasma degradation of AOH and AME was illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig15\" class=\"InternalRef\"\u003e5\u003c/span\u003e. Even when exposed to varying concentrations of oleic acid, AOH could be completely degraded after a 3-min treatment with CP. For AME, the degradation percentage was 95.5%, 93.7%, and 96.9% at oleic acid concentrations of 0.0500, 0.100, and 0.150 mg/mL respectively; however, complete degradation occurred within 5 min regardless of concentration level. These results demonstrated that oleic acid had little effect on the degradation of AOH and AME by CP. Figure\u0026nbsp;\u003cspan refid=\"Fig15\" class=\"InternalRef\"\u003e5\u003c/span\u003e also depicted the effect of linoleic acid on plasma degradation of AOH and AME; no significant impact was observed for AOH while slight effects were noted for AME at the highest concentration tested (0.15 mg/mL) under these experimental conditions.\u003c/p\u003e \u003cp\u003eBatista et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) demonstrated that the fatty acid profile remains unaffected by CP technology. Perez-Andres et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) investigated the impact of cold atmospheric plasma treatment on commercially packaged mackerel fillets and reported insignificant alterations in both fatty acid composition and nutritional quality indices post-treatment. The fact that the degradation of mycotoxin by CP was not significantly affected in the presence of oleic acid and linoleic acid might be because oleic acid and linoleic acid did not consume the active substances produced by CP.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eMechanism of the effect of jujube components on mycotoxin plasma degradation\u003c/h2\u003e \u003cp\u003e \u003cb\u003eThe effect of jujube components on the electrochemical properties of the systems after cold plasma discharge\u003c/b\u003e \u003c/p\u003e \u003cp\u003eORP is a measure of a solution's ability to oxidize or reduce another substance, reflecting the overall level of RONS present in the solution. A higher ORP value indicates a stronger oxidation capacity (Thirumdas et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). As shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig16\" class=\"InternalRef\"\u003e6\u003c/span\u003e, plasma treatment resulted in varying degrees of increase in the ORP of the solution. The results of this experimental study demonstrated that the changes in △ORP for jujube protein solution, jujube polysaccharide, and simulated jujube juice were lower compared to an aqueous solution. However, solutions with different concentrations of Vc, K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e2+\u003c/sup\u003e, oleic acid, and linoleic acid exhibited similar changes in △ORP as observed for an aqueous solution. The variation trend of △ORP was highly consistent with the degradation pattern of toxins, suggesting that the effect of jujube juice components on AOH and AME degradation might be attributed to the oxidative capability of certain RONS generated by CP discharge.\u003c/p\u003e \u003cp\u003eH\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, a major active substance generated during CP discharge, exhibits strong oxidation effects (Ma et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). As depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig17\" class=\"InternalRef\"\u003e7\u003c/span\u003e, the H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e content of solutions containing various jujube components such as jujube protein, jujube polysaccharide, Ca\u003csup\u003e2+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, Vc, oleic acid, and linoleic acid was significantly lower than that of aqueous solution after CP treatment. Tripathi and Mishra (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) suggested that H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e can degrade mycotoxins via oxidation. Additionally, Ma et al. (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) highlighted the crucial role of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e in chitosan degradation during CP discharge and its decomposition to produce \u0026bull;OH. The impact of different jujube juice components on AOH and AME degradation by CP might be attributed to their partial consumption of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e.\u003c/p\u003e \u003cp\u003eThe effect of CP on the \u0026bull;OH scavenging rate of various jujube juice component model systems was shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig17\" class=\"InternalRef\"\u003e7\u003c/span\u003e. The results showed that with the increase of jujube protein concentration from 0 g/mL to 0.00750 g/mL, the \u0026bull;OH clearance decreased significantly from 70.3\u0026ndash;24.1%. Furthermore, an increase in jujube polysaccharide concentration to 0.0750 g/mL resulted in a reduction of \u0026bull;OH clearance decreased by 6.19%. Conversely, no notable change was observed in \u0026bull;OH clearance following CP treatment across different concentrations of linoleic acid, Ca\u003csup\u003e2+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, and Vc environments. As for oleic acid, there was an initial significant decrease followed by an increase in \u0026bull;OH clearance with increasing concentration. Notably, compared to the aqueous solution, the multi-component model system of jujube juice exhibited a substantial decrease in \u0026bull;OH clearance with increasing concentration. It is worth mentioning that \u0026bull;OH is one of the primary short-lived reactive species generated during CP discharge (Kurake et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Several scholars have argued that the oxidizing effectors produced by CP are primarily attributed to \u0026bull;OH radicals which can disrupt structurally important bonds within peptidoglycan (i.e., C-O, C-N, or C-C bonds) (Yusupov et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Moreover, variations observed in \u0026bull;OH clearance were highly consistent with the degradation effects exerted by AOH and AME under different jujube component model systems; thus indicating a strong correlation between their removal from jujube juice through CP treatment and \u0026bull;OH generation during discharge.\u003c/p\u003e \u003cp\u003eIn addition to ROS, CP discharge also generates extremely rich RNS (NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, etc.) (Sardella et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Yusupov et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Lukes et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Therefore, it is imperative to investigate the impact of CP on NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e in various model systems representing jujube juice components (Fig.\u0026nbsp;\u003cspan refid=\"Fig18\" class=\"InternalRef\"\u003e8\u003c/span\u003e). The results revealed that following CP treatment, the concentration of NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e significantly decreased for different jujube juice components compared to the aqueous solution. This reduction could be attributed to two factors. Firstly, diverse jujube juice components might consume some NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e through chemical reactions with them. Secondly, there might be a conversion of NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e into NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. Luke et al. (2014) proposed that in aqueous solutions, unstable NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e could undergo transformation into more stable form i.e., NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, leading to pH decrease and increased acidity.\u003c/p\u003e \u003cp\u003eThe impact of CP on the NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e content in various jujube juice component model systems was illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig18\" class=\"InternalRef\"\u003e8\u003c/span\u003e. Following an equivalent duration of CP treatment, distinct jujube juice component solutions exhibited varying levels of NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e content, with significantly lower concentrations observed when exposed to jujube polysaccharide and protein compared to aqueous solution. However, the presence of Ca\u003csup\u003e2+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, oleic acid, linoleic acid, and Vc had minimal influence on the NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e concentration generated by CP. Consequently, the degradation of AOH and AME by CP in jujube juice was found to be closely associated with the production of NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e during CP discharge.\u003c/p\u003e \u003cp\u003e \u003cb\u003eThe correlation analysis between the changes of electrochemical properties of the system and the CP degradation of toxins\u003c/b\u003e \u003c/p\u003e \u003cp\u003eAfter CP treatment, the Pearson correlation analysis revealed in Fig.\u0026nbsp;\u003cspan refid=\"Fig19\" class=\"InternalRef\"\u003e9\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig20\" class=\"InternalRef\"\u003e10\u003c/span\u003e demonstrated that the degradation of AOH and AME was significantly positively correlated with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;,\u003c/sup\u003e and ORP while being negatively correlated with pH when exposed to jujube protein and polysaccharide. Furthermore, a significant correlation was observed between the physical and chemical properties as well as active substance content of jujube protein and polysaccharide solution after CP treatment. This suggested that the inhibition of toxin degradation by jujube protein and polysaccharide was closely related to changes in physicochemical properties and active substances produced by CP.\u003c/p\u003e \u003cp\u003eIn the Pearson correlation analysis of AOH and AME degradation and the changes in Vc model system properties, the degradation trend of AOH exhibited significant positive correlations with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e contents, while showing a negative correlation with pH value. However, no significant correlations were observed between △ORP and conductivity. Meanwhile, the degradation trend of AME showed positive correlations with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e content, NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e content, and △ORP; whereas it displayed a negative correlation with pH.\u003c/p\u003e \u003cp\u003eRegarding the Ca\u003csup\u003e2+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, oleic acid, and linoleic acid model systems, the degradation trend of AOH showed only partial or insignificant correlation with certain properties or active substances. In summary, following CP treatment, there was a significant positive correlation between the change in △ORP, the contents of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e of jujube polysaccharide, jujube protein, and multi-component model system and the degradation trend of toxins. Additionally, the degradation trend of toxins in Vc system exhibited a high correlation with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. However for oleic acid, linoleic acid, Ca\u003csup\u003e2+\u003c/sup\u003e, and K\u003csup\u003e+\u003c/sup\u003e, the degradation trend of toxins displayed less correlation with the properties of these systems. Furthermore, the results from Pearson correlation analysis further demonstrated that jujube components could influence the degradation of AOH and AME by CP through consumption of active particles including H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e. It can be inferred that RONS produced by CP is considered as a primary factor responsible for toxin degradation.\u003c/p\u003e \u003cp\u003eOverall, the present study demonstrated the effective degradation of AOH and AME in both aqueous solution and jujube juice by CP. However, the degradation was partially inhibited in jujube juice. Among these tested components of jujube juice, jujube protein exhibited the highest inhibitory effect on plasma degradation of these two toxins, followed by jujube polysaccharide. Other components including Vc, Ca\u003csup\u003e2+\u003c/sup\u003e, K\u003csup\u003e+\u003c/sup\u003e, oleic acid, or linoleic acid showed insignificant effects on AOH and AME degradation. Furthermore, the degradation of AOH and AME by CP was closely associated with the presence of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e active particles in different discharge modes during CP treatment. After CP treatment, consumption of RONS such as H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, \u0026bull;OH, NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e, and NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e by jujube protein or polysaccharide may be a key factor contributing to inhibition of AOH and AME degradation. This study provides a theoretical basis for understanding the mechanism underlying \u003cem\u003eAlternaria\u003c/em\u003e mycotoxin degradation in jujube juice system using CP technology and promotes its application for efficient targeted detoxification in food systems.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003eWe appreciate the help of Key Scientific Research Projects of Colleges and Universities in Henan province (24A550020) established by Department of Education of Henan Province.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCRediT authorship contribution statement\u0026nbsp;\u003c/strong\u003eX. Y. W. : Investigation, methodology, writing, review \u0026amp; editing, funding acquisition, resources. Q. L. : methodology, formal analysis. Y. K. H. : Investigation, methodology, formal analysis. Z. Z. G. : review \u0026amp; editing. X. P. W. : Writing, review \u0026amp; editing. Y. Z. Y. : methodology, formal analysis. W. Z. : Supervision, review \u0026amp; editing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e This research was supported by Key Scientific Research Projects of Colleges and Universities in Henan province (24A550020) established by Department of Education of Henan Province.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAttri, P., Kim, Y.H., Park, D.H., Park, J.H., Hong, Y.J., Uhm, H.S., Kim, K.-N., Fridman, A. and Choi, E.H., 2015. Generation mechanism of hydroxyl radical species and its lifetime prediction during the plasma-initiated ultraviolet (UV) photolysis. 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European Physical Journal Special Topics 226: 2901-2910. https://doi.org/10.1140/epjst/e2016-60331-4\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Alternaria mycotoxins, alternariol (AOH), alternariol monomethyl ether (AME), cold plasma, mycotoxin degradation, jujube juice components","lastPublishedDoi":"10.21203/rs.3.rs-4146628/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4146628/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe study examined the impact of jujube constituents on cold plasma (CP) degradation of alternariol (AOH) and alternariol monomethyl ether (AME), followed by assessing changes in CP electrochemical properties. The correlation between these properties and toxin degradation was analyzed to understand how jujube components influence the breakdown of AOH and AME. Results showed that when treated with CP for 3 minutes, 0.00350 mg/mL jujube protein degraded 49.0% of AOH and 48.8% of AME. Under the same treatment time, 0.0500 mg/mL jujube polysaccharide degraded AOH by 74.6% and AME by 95.8%. Conversely, Vc, K\u003csup\u003e+\u003c/sup\u003e, Ca\u003csup\u003e2+\u003c/sup\u003e, oleic acid, and linoleic acid exhibited negligible inhibitory effects on toxin degradation. Furthermore, following CP treatment, oxidation-reduction potential (△ORP), pH, conductivity, hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e), hydroxyl radical (\u0026bull;OH), nitrate ion (NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e), and nitrite ion (NO\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e) contents in the jujube protein or polysaccharide system were significantly correlated with toxin degradation; thus confirming their association with reactive oxygen-nitrogen species (RONS). The presence of jujube protein or polysaccharide hindered the degradation of CP-mediated toxins by consuming reactive RONS resources. This study provides insights into how AOH and AME in jujube juice are degraded by CP, enabling more targeted and efficient elimination of foodborne toxins.\u003c/p\u003e","manuscriptTitle":"Effects and mechanisms of jujube juice components on degradation of Alternaria mycotoxin by cold plasma","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-28 09:17:35","doi":"10.21203/rs.3.rs-4146628/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8b5cec9f-fcd2-4124-b9e3-d13c7f292fc9","owner":[],"postedDate":"March 28th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-04-01T06:05:16+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-28 09:17:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4146628","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4146628","identity":"rs-4146628","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}