Citric Acid Research Articles

Acetic and citric acids effect the type II secretion system and decrease the metabolic activities of salmon spoilage-related Rahnella aquatilis KM05

Rahnella aquatilis causes seafoods to spoil by metabolizing sulfur-containing amino acids and/or proteins, producing H2S in products. The type II secretion system (T2SS) regulates the transport of proteases from the cytoplasm to the surrounding environment and promotes bacterial growth at low temperatures. To prevent premature fish spoilage, new solutions for inhibiting the T2SS of bacteria should be researched. In this study, global transcriptome sequencing was used to analyze the spoilage properties of R. aquatilis KM05. Two of the mapped genes/coding sequences (CDSs) were matched to the T2SS, namely, qspF and gspE, and four of the genes/CDSs, namely, ftsH, rseP, ptrA and pepN, were matched to metalloproteases or peptidases in R. aquatilis KM05. Subinhibitory concentrations of citric (18 µM) and acetic (41 µM) acids caused downregulation of T2SS-related genes (range from − 1.0 to -4.5) and genes involved in the proteolytic activities of bacteria (range from − 0.5 to -4.0). The proteolytic activities of R. aquatilis KM05 in vitro were reduced by an average of 40%. The in situ experiments showed the antimicrobial properties of citric and acetic acids against R. aquatilis KM05; the addition of an acidulant to salmon fillets limited microbial growth. Citric and acetic acids extend the shelf life of fish-based products and prevent food waste.

Low doses of acetyl trihexyl citrate plasticizer promote adipogenesis in hepatocytes and mice.

Accumulating epidemiological evidence underscores the association between pervasive environmental factors and an increased risk of metabolic diseases. Environmental chemicals, recognized disruptors of endocrine and metabolic processes, may contribute to the global prevalence of metabolic disorders, including obesity. Acetyl tributyl citrate (ATHC), categorized as a citric acid ester plasticizer, serves as a substitute for di-(2-ethylhexyl) phthalate (DEHP) in various everyday products. Despite its widespread use and the increasing risk of exposure in humans and animals due to its high leakage rates, information regarding the safety of exposure to environmentally relevant doses of ATHC remains limited. This study aimed to investigate the potential impact of ATHC exposure on metabolic homeostasis. Both in vivo and in vitro exposure models were used to characterize the effects induced by ATHC exposure. C57BL/6J male mice were subjected to a diet containing ATHC for 12weeks, and metabolism-related parameters were monitored and analyzed throughout and after the exposure period. Results indicated that sub-chronic dietary exposure to ATHC induced an increase in body fat percentage, elevated serum lipid levels, and increased lipid content in the liver tissue of mice. Furthermore, the effect of ATHC exposure on murine hepatocytes were examined and results indicated that ATHC significantly augmented lipid levels in AML12 hepatocytes, disrupting energy homeostasis and altering the expression of genes associated with fatty acid synthesis, uptake, oxidation, and secretion pathways. Conclusively, both in vivo and in vitro results suggest that exposure to low levels of ATHC may be linked to an elevated risk of obesity and fatty liver in mice. The potential implications of ATHC on human health warrant comprehensive evaluation in future studies.

Innovative Wearable Sweat Sensor Array for Real-Time Volatile Organic Compound Detection in Noninvasive Diabetes Monitoring.

Wearable sweat sensors are reshaping healthcare monitoring, providing real-time data on hydration and electrolyte levels with user-friendly, noninvasive devices. This paper introduces a highly portable two-channel microfluidic device for simultaneous sweat sampling and the real-time detection of volatile organic compound (VOC) biomarkers. This innovative wearable microfluidic system is tailored for monitoring diabetes through the continuous and noninvasive tracking of acetone and ammonia VOCs, and it seamlessly integrates with smartphones for easy data management. The core of this system lies in the utilization of carbon polymer dots (CPDs) and carbon dots (CDs) derived from monomers such as catechol, resorcinol, o-phenylenediamine, urea, and citric acid. These dots are seamlessly integrated into hydrogels made from gelatin and poly(vinyl alcohol), resulting in an advanced solid-state fluorometric sensor coating on a cellulose paper substrate. These sensors exhibit exceptional performance, offering linear detection ranges of 0.05-0.15 ppm for acetone and 0.25-0.37 ppm for ammonia, with notably low detection limits of 0.01 and 0.08 ppm, respectively. Rigorous optimization of operational parameters, encompassing the temperature, sample volume, and assay time, has been undertaken to maximize device performance. Furthermore, these sensors demonstrate impressive selectivity, effectively discerning between biologically similar substances and other potential compounds commonly present in sweat. As this field matures, the prospect of cost-effective, continuous, personalized health monitoring through wearable VOC sensors holds significant potential for overcoming barriers to comprehensive medical care in underserved regions. This highlights the transformative capacity of wearable VOC sweat sensing in ensuring equitable access to advanced healthcare diagnostics, particularly in remote or geographically isolated areas.

Integrative Proteomics-Metabolomics of In Vitro Degeneration of Cardiovascular Cell Lines.

Long-term cell culture is an important biological approach but is also characterized by degeneration in cellular morphology, proliferation rate, and function. To explore this phenomenon in a systematic way, we conducted an integrative proteomics-metabolomics measurement of two cardiovascular cell lines of AC16 and HUVECs. The 18th culturing passages, i.e., G18, showed as the turning points by cell metabolism profiles, in which the metabolomic changes demonstrated the dysfunction of energy, amino acid, and ribonucleotide metabolism metabolic pathways. Although active protein networks showed mitochondria abundance AC16 and oxidative/nitrative sensitive HUVECs indicated the different degeneration patterns, the G18 and G30 proteomics evidenced the senescence by processes of signal transduction, signaling by interleukins, programmed cell death, cellular responses to stimuli, cell cycle, mRNA splicing, and translation. Some crucial proteins (RPS8, HNRNPR, SOD2, LMNB1, PSMA1, DECR1, GOT2, OGDH, PNP, CBS, ATIC, and IMPDH2) and metabolites (L-glutamic acid, guanine, citric acid, guanosine, guanosine diphosphate, glucose 6-phosphate, and adenosine) that contributed to the dysregulation of cellular homeostasis are identified by using the integrative proteomic-metabolomic analysis, which highlighted the increased cellular instability. These findings illuminate some vital molecular processes when culturing serial passages, which contribute holistic viewpoints of in vitro biology with emphasis on the replicative senescence of cardiovascular cells.

A universal strategy towards high-rate and ultralong-life of Li‐rich Mn‐based cathode materials

Lithium-rich manganese-based cathode materials (LRMs) have shown promise for the next-generation lithium battery cathodes due to their high discharge specific capacity (∼250 mAh g−1) and wide operating voltage range (2.0–4.8 V). However, these materials face limitations such as low initial Coulombic efficiency (ICE), poor cycle stability, and inadequate rate capability. To address these challenges, we employed a simple citric acid treatment (CA-treatment) method to fabricate the Li-rich spinel coating layer on LRMs. This in situ formed spinel Li4Mn5O12 layer successfully suppresses the oxygen release, provides three-dimensional (3D) lithium-ion diffusion channels and enriches Li embedding sites, resulting in a substantial improvement in the rate capability and high-rate cycling performance. The modified LRM delivers a high specific capacity of 193.7 mAh g−1 at 2.0 C and 154.8 mAh g−1 at 5.0 C, after 1000 cycles at 10.0 C, 77.4 % capacity retention with 102.5 mAh g−1 is realized. This facile CA-treatment benefits the electrochemical properties of LRMs comparable to those of altering the microscopic morphology. This work offers a new avenue for exploring outstanding LRMs with high-rate and ultralong cycling life.

The study of citric acid crosslinked β-cyclodextrin/hydroxypropyl cellulose food preservation film

Abstract In this study, hydroxypropyl cellulose (HPC) was utilized as the raw material, with the addition of β-cyclodextrin (β-CD), citric acid (CA) as the crosslinking agent, and sodium hypophosphite (SHP) as the catalyst to produce hydroxypropyl cellulose/β-CD composite films. The inclusion of β-CD resulted in an increase in the tensile strength of the film, with the maximum value of 13.5 MPa for the 1 % β-CD composite membrane. Additionally, after degradation in soil for 28 days, the degradation ability was significantly enhanced, with the 1.0 % β-CD composite film exhibiting the highest degradation rate of 27.21 %. Furthermore, the water permeability of the composite membrane was improved with the addition of β-CD. Specifically, when the β-CD content was 1.0 %, the water vapor transmission reached its lowest point at 2,445 g* ( m 2 * 24 d ) − 1 ${({m}^{2}\ast 24d)}^{-1}$ . The findings demonstrated that the 1 % β-cyclodextrin/hydroxypropyl cellulose composite film effectively preserved the freshness of strawberries, reducing the weight loss rate by 1.65 % compared to the control group. In conclusion, this research highlights the potential for preparing composite membranes using HPC and β-CD crosslinking, thereby expanding the application of hydroxypropyl cellulose and β-CD in food preservation.

Tradeoffs among plant yield, nitrate accumulation risk, and potential pathogen: Effects of citric acid and nitrification inhibitors on soil–plant systems

Tradeoffs among plant yield, nitrate accumulation risk, and potential pathogen: Effects of citric acid and nitrification inhibitors on soil–plant systems

Assessing a respiratory toxic infectious bronchitis virus (IBV) strain: isolation, identification, pathogenicity, and immunological failure insights.

Infectious bronchitis virus (IBV) is caused by avian coronavirus and poses a global economic threat to the poultry industry. In 2023, a highly pathogenic IBV strain, IBV/CN/GD20230501, was isolated and identified from chickens vaccinated with IBV-M41 in Guangdong, China. This study comprehensively investigated the biological characteristics of the isolated IBV strain, including its genotype, whole genome sequence analysis of its S1 gene, pathogenicity, host immune response, and serum non-targeted metabolomics. Through the analysis of the S1 gene sequence, serum neutralization tests, and comparative genomics, it was proven that IBV/CN/GD20230501 belongs to the GI-I type of strain and is serotype II. One alanine residue in the S1 subunit of the isolated strain was mutated into serine, and some mutations were observed in the ORF1ab gene and the terminal region of the genome. Animal challenge experiments using the EID50 and TCID50 calculations showed that IBV/CN/GD20230501 possesses strong respiratory pathogenicity, with early and long-term shedding of viruses and rapid viral spread. Antibody detection indicated that chickens infected with IBV/CN/GD20230501 exhibited delayed expression of early innate immune genes, while those infected with M41 showed rapid gene induction and effective viral control. Metabolomics analysis demonstrated that this virus infection led to differential expression of 291 ions in chicken serum, mainly affecting the citric acid cycle (tricarboxylic acid cycle).IMPORTANCEThis study identified an infectious bronchitis virus (IBV) strain isolated from vaccinated chickens in an immunized population that had certain sequence differences compared to IBV-M41, resulting in significantly enhanced pathogenicity and host defense. This strain has the potential to replace M41 as a more suitable challenge model for drug research. The non-targeted metabolomics analysis highlighting the citric acid cycle provides a new avenue for studying this highly virulent strain.

Genome-wide association studies for citric and lactic acids in dairy sheep milk in a New Zealand flock

The objectives of this study were to estimate genetic parameters for citric acid content (CA) and lactic acid content (LA) in sheep milk and to identify the associated candidate genes in a New Zealand dairy sheep flock. Records from 165 ewes were used. Heritability estimates based on pedigree records for CA and LA were 0.65 and 0.33, respectively. The genetic and phenotypic correlations between CA and LA were strong-moderate and negative. Estimates of genomic heritability for CA and LA were also high (0.85, 0.51) and the genomic correlation between CA and LA was strongly negative (-0.96 ± 0.11). No significant associations were found at the Bonferroni level. However, one intragenic SNP in C1QTNF1 (chromosome 11) was associated with CA, at the chromosomal significance threshold. Another SNP associated with CA was intergenic (chromosome 15). For LA, the most notable SNP was intragenic in CYTH1 (chromosome 11), the other two SNPs were intragenic in MGAT5B and TIMP2 (chromosome 11), and four SNPs were intergenic (chromosomes 1 and 24). The functions of candidate genes indicate that CA and LA could potentially be used as biomarkers for energy balance and clinical mastitis. Further research is recommended to validate the present results.

Effect of Rheum ribes L. pulp enriched with eugenol or thymol on survival of foodborne pathogens and quality parameters of chicken breast fillets

The aim of this study was to characterize the pulp of Rheum ribes L. and to determine the effect of the pulp enriched with eugenol (1 %) or thymol (1 %) on the microbiological and physico-chemical quality of chicken breast fillets. Chicken breast fillets, inoculated with Listeria monocytogenes, Salmonella enterica subsp. enterica serovar Typhimurium, and Escherichia coli O157:H7 (~6.0 log10), were marinated for 24 h in a mixture prepared from a combination of Rheum ribes L. pulp with eugenol or thymol. The quality parameters were analyzed for 15 days at +4 °C. The Rheum ribes L. pulp was found to have high antioxidant activity, high total phenolic content and contained 22 different phenolic substances, among which rutin ranked first. The pulp contained high levels of p-xylene and o-xylene as volatile substances and citric acid as an organic acid. The combination of Pulp + Eugenol + Thymol (PET) reduced the number of pathogens in chicken breast fillets by 2.03 to 3.50 log10 on day 0 and by 2.25 to 4.21 log10 on day 15, compared to the control group (P < 0.05). The marinating treatment significantly lowered the pH values of fillet samples on the first day of the study, compared to the control group (P < 0.05). During storage, TVB-N levels showed slower increase in the treatment groups compared to the control group (P < 0.05). In addition, the marinating process led to significant changes in physicochemical parameters such as water holding capacity, color, texture, cooking loss, and drip loss compared to the control group (P < 0.05). In conclusion, the results of this study showed that the pulp of Rheum ribes L., which has a high antioxidant capacity and contains various bioactive compounds. Furthermore, S. Typhimurium, E. coli O157:H7 and L. monocytogenes were inhibited considerably by marinating Rheum ribes L. pulp with a combination of eugenol and thymol.

Dehydrosilylation of Alcohols Using Gold Nanoparticles Deposited on Citric Acid-modified Fibrillated Cellulose

The development of an effective catalytic system for the dehydrogenative coupling of hydrosilanes with alcohols remains an ongoing challenge, particularly for alcohol protection applications. In this study, we report the development and optimization of a highly efficient gold catalyst supported on fibrillated cellulose modified with citric acid (F-CAC). The catalyst exhibited remarkable catalytic activity under mild conditions with 0.01–0.05 mol% of Au loading, facilitating the formation of silyl ethers with excellent yield. Notably, our catalytic system overcomes the need for excess alcohol typically required in such reactions, making it highly practical for alcohol protection applications. This work represents a significant advancement in the field of dehydrosilylation catalysis, offering a sustainable, efficient, and environmentally friendly approach for the synthesis of functional silanol-based materials and alcohol protection applications. The scope of substrates and the utility of the catalyst have been thoroughly studied.

Padina gymnospora biorefinery: coalescing ultrasonic-assisted phenolic recovery and citric acid–mediated alginate extraction

Padina gymnospora biorefinery: coalescing ultrasonic-assisted phenolic recovery and citric acid–mediated alginate extraction

Deciphering Blood Flow Restriction Training to Aid Lipid Lowering in Obese College Students through Untargeted Metabolomics

(1) Objective: The aim of this study was to observe the lipid-lowering effects of blood flow restriction training (BFR) combined with moderate-intensity continuous training (MICT) in obese college students by observing lipid-lowering hormones and untargeted metabolomics. (2) Methods: In this study, 14 obese college students were convened into three groups—MICT, MICT+BFR, and high-intensity interval training (HIIT)—for a crossover experiment. Blood was drawn before and after exercise for the analysis of lipolytic agents and untargeted metabolomics. The study used a paired t-test and ANOVA for statistical analyses. (3) Results: The lipolytic agent results showed that MICT+BFR was superior to the other two groups in terms of two agents (p = 0.000 and p = 0.003), namely, GH and IL-6 (difference between before and after testing: 10,986.51 ± 5601.84 and 2.42 ± 2.49, respectively), and HIIT was superior to the other two groups in terms of one agent (p = 0.000), i.e., EPI (22.81 ± 16.12). No advantage was observed for MICT. The metabolomics results showed that, compared to MICT, MICT+BFR was associated with the upregulated expression of xanthine, succinate, lactate, N-lactoylphenylalanine, citrate, ureido acid, and myristic acid after exercise, with the possibility of the involvement of the citric acid cycle, alanine, aspartic acid, glutamate metabolism, butyric acid metabolism, and the histidylate metabolism pathway. (4) Conclusions: The superior lipid-lowering effect of MICT+BFR over MICT in a group of obese college students may be due to the stronger activation of GH and IL-6 agents, with the citric acid cycle and alanine, aspartate, and glutamate metabolic pathways being associated with this type of exercise.

Antibacterial characteristics and mechanistic insights of combined tea polyphenols, Nisin, and epsilon-polylysine against feline oral pathogens: a comprehensive transcriptomic and metabolomic analysis.

This study evaluates the antibacterial characteristics and mechanisms of combined tea polyphenols (TPs), Nisin, and ε-polylysine (PL) against Streptococcus canis, Streptococcus minor, Streptococcus mutans, and Actinomyces oris, common zoonotic pathogens in companion animals. Pathogenic strains were isolated from feline oral cavities and assessed using minimum inhibitory concentration (MIC) tests, inhibition zone assays, growth kinetics, and biofilm inhibition studies. Among single agents, PL exhibited the lowest MIC values against all four pathogens. TP showed significant resistance against S. minor, and Nisin against S. mutans. The combination treatment (Comb) of TP, Nisin, and PL in a ratio of 13:5:1 demonstrated broad-spectrum antibacterial activity, maintaining low MIC values, forming large inhibition zones, prolonging the bacterial lag phase, reducing growth rates, and inhibiting biofilm formation. RNA sequencing and metabolomic analysis indicated that TP, Nisin, and PL inhibited various membrane-bound carbohydrate-specific transferases through the phosphoenolpyruvate-dependent phosphotransferase system in S. canis, disrupting carbohydrate uptake. They also downregulated glycolysis and the citric acid cycle, inhibiting cellular energy metabolism. Additionally, they modulated the activities of peptidoglycan glycosyltransferases and d-alanyl-d-alanine carboxypeptidase, interfering with peptidoglycan cross-linking and bacterial cell wall stability. The Comb therapy significantly enhances antibacterial efficacy by targeting multiple bacterial pathways, offering potential applications in food and pharmaceutical antimicrobials.

Effect of Low Molecular Weight Organic Acids and Liming on Phosphorus Adsorption in Acidic Tea Soils in the Black Sea Region, Türkiye

ABSTRACT Agricultural productivity is significantly hindered by the limited availability of phosphorus in acidic soils due to the presence of aluminum and iron phosphates. Soil liming is commonly employed to enhance soil fertility. However, modifying the pH might also impact the interactions involving existing soil organic matter, leading to changes in the effective concentrations of competing low-molecular-weight acids (LMWOACs) participating in phosphorus adsorption. This study explored the impacts of citric, oxalic, and malic acids in acidic soils from tea cultivation regions within the Eastern Black Sea. These soils had undergone various levels of liming and were incubated for 15 months to assess their effects on phosphorus (P) adsorption in 7 acidic soils. The experimental sorption data were utilized to calculate the soil Langmuir and Freundlich adsorption parameters. In addition, the influences of organic acids and liming levels on the Freundlich and Langmuir model parameters were subjected to ANOVA analysis. The findings indicated that citric acid demonstrated the highest efficiency in diminishing phosphorus adsorption among the tested organic acids, followed by oxalic and malic acids. Three carboxyl groups (−COOH) in citric acid facilitated the formation of more complexes with adsorbed phosphorus on the soil surface. Furthermore, the 100% LR ratio of liming notably decreased phosphorus adsorption, especially in the lower end of the isotherms. Therefore, it can be concluded that the combination of LMWOACs and liming treatments could be used to overcome P deficiency in acid soils with very high P sorption ability.

Physicochemical and phytochemical characterization of opuntia dillenii: A promising source of bioactive compounds

ABSTRACT The objective of this ongoing research is to investigate and assess the aqueous extracts, vegetable oil, and residual plant components derived from O. dillenii by use of HPLC, and GC-MS. Subsequent investigations employed colorimetric techniques to identify reducing and soluble sugars. We developed and validated an energy-dispersive X-ray fluorescence (EDXRF) method for determining the concentrations of essential chemical elements (O, C, Ca, Mg, K, S, P) in seeds, juice, and peel, as well as their residues. HPLC analysis yielded results indicating that extracts from O. dillenii seeds, juice, and peel from distinct regions (Oujda, Nador, and Essaouira) contained elevated concentrations of metabolites. Specifically, peel and juice extracts exhibited the highest levels of organic acids and betanin compared to O. dillenii seeds. Conversely, seed oils displayed a noteworthy tocopherol content, predominantly δ-tocopherol. Similarly, an examination of organic acid composition in juice, peel, and seeds from three different regions revealed their elevated levels. Citric acid and oxalic acid emerged as predominant components in the juice. Overall, Opuntia dillenii’s physicochemical and phytochemical characterization improves in understanding its potential as a bioactive chemical promoter, which could lead to the creation of new medicinal medicines or functional food ingredients.

Evaluating the performance of citric acid and maleic acid for mixed-acid leaching of critical metals from spent lithium-ion batteries

Evaluating the performance of citric acid and maleic acid for mixed-acid leaching of critical metals from spent lithium-ion batteries

Delayed ammonia release of ammonia borane hydrolysis by citric acid

Ammonia borane (AB), with high hydrogen density and stability under ambient conditions, readily releases hydrogen via catalyzed hydrolysis. However, ammonia, a byproduct of hydrogen release, limits its utilization. Citric acid, a safe and cost-effective acid catalyst, for the acid catalyst of AB hydrolysis was investigated in this study to mitigate ammonia emissions while enhancing hydrogen release. Maximal hydrogen release (3 eq.H2) and ammonia emission suppression (<0.5 ppm) were achieved using a 2-mol/L AB aqueous solution adding into citric acid powder (molar ratio of AB: citric acid = 1:1). In contrast, the amount of hydrogen release at other concentrations and/or using the mixing method was below 2.7 mol/mol AB within several hours. Ammonia re-released even after hydrogen release in the case of high AB concentration (e.g. >4 mol/L AB aqueous solution with citric acid powder (molar ratio of AB: citric acid = 1:1)) in the solution because of formation of borate ester (1:2 borate–diol molar ratio) via mono-ammonium citrate (a borate-ester precursor), which was revealed by the solution NMR. These results reveal a cost-effective and simple on-way method for hydrogen release with suppressed impurities. Furthermore, understanding impurity generation mechanisms can contribute to utilizing AB-derived hydrogen.

Phytochemical Characterization and Bioactivity of Extracts from Different Fruit Parts of Opuntia leucotricha DC.: A Comparison between a Conventional Organic Solvent and Green Natural Deep Eutectic Solvents

The objective of this study was to analyze the chemical profile of extracts from different fruit parts of the perennial arborescent cactus Opuntia leucotricha by applying ultra-high-performance liquid chromatography–high-resolution mass spectrometry (UHPLC-HRMS) and to evaluate their biological properties (antioxidant and enzyme inhibitory capacities). Extracts were obtained from the fruit pulp (without or with seeds), seeds, and peel by ultrasound-assisted extraction (UAE) using the conventional solvent methanol 50% and two Natural Deep Eutectic Solvents (NADESs) (glycerol:urea, 1:1 and citric acid:sorbitol, 1:2). A total of 33 compounds were identified, including phenolics, fatty acids, and others. Phenolic acids were the most abundant class of phenolics identified in all fruit parts, with the highest concentration observed in the methanol extracts from peel (593.02 µg/gDW), followed by seed-containing pulp (69.03 µg/gDW), pulp (57.83 µg/gDW), and seeds (39.97 µg/gDW). The second most effective extractant was NADES 1 (glycerol:urea, 1:1), which was also successful in extracting compounds with antioxidant capacity. Overall, the extracts demonstrated considerable enzyme inhibitory activities, with the greatest effects observed against α-amylase and α-glucosidase. The results indicate that O. leucotricha fruits could be a promising source of bioactive compounds, and NADES a viable alternative to organic solvents for their industrial exploitation.

Stability optimization of iron-based catalysts: Application of Cr-enriched alloy steel catalyst in ozonation pharmaceutical wastewater

Catalytic ozonation for advanced treatment of industrial wastewater has proved to be effective, and the stability of the catalyst is crucial for the long-term application of the ozonation process in practical engineering. In this study, the electrochemical characteristics of four iron (Fe) shavings: alloy steels (38CrMoAl and 40Cr) and carbon steels (A3 and 45#) were investigated, and a new citric acid-optimized H2O2 modification method was proposed. The low open circuit potential (OCP) of carbon steels indicated that they were more likely to be oxidized therefore required shorter modification times. The higher the OCP of alloy steels, the higher H2O2 concentration was required to enhance modification efficiency. By adding citric acid, the rapid modification of alloy steel was achieved, and the catalysts exhibited lower passivation current (Ip), indicating a lower rate of substrate loss and a longer lifetime of alloy steel catalysts. Furthermore, chromium (Cr) enrichment occurred in the oxide layer of alloy steel catalysts. The Cr/Fe (atomic ratio) in oxide layer for 38CrMoAl and 40Cr increased from 0.018 to 0.040 and from 0.010 to 0.037, respectively. The presence of CrOOH and Cr2O3 within the oxide layer reinforced the layer’s overall structural integrity and enhanced its protection for the internal substrate, making the catalysts more resistant to adapt to high-salinity wastewater. Through laboratory tests, it was found that among the four catalysts, 38CrMoAl showed the highest stability and good catalytic activity with low Fe loss concentration. Therefore, the pilot-scale application effect of 38CrMoAl catalyst was evaluated. In the advanced treatment of pharmaceutical wastewater, chemical oxygen demand (COD) and total organic carbon (TOC) removals were 56.3% and 32.4%, respectively. These results showed that the stability of catalyst can be improved by optimizing the selection of steel, and the introduction of citric acid achieved Cr enrichment effect and amplified the potential stability advantage of alloy steel. The research provides guidance on raw material selection for catalyst preparation to different industries' wastewater treatment, especially in high-salinity wastewater ozonation treatment, which can realize the long-term stable application of catalyst.