A hyperglycemic condition, characterized by high glucose concentration in the bloodstream, is one of the most potent dietary risk factors for colorectal cancer (CRC). It regulates the response to oxidative stress, migration, and invasion of CRC cells, as well as affects the integrity of the intestinal barrier, which plays a pivotal role in protecting against chemicals and contaminants present in the diet. Although the molecular mechanisms underlying the toxicity of the mycotoxins deoxynivalenol (DON) and alternariol (AOH) are quite well-known, no research so far has considered the possibility that high glucose concentration might affect the response of CRC cells to mycotoxins. In this study, we evaluated how high glucose condition (50 mmol/L) modulates the effect of two mycotoxins distinct in toxicity and underlying mechanisms: DON (0.1-1 μmol/L) and AOH (0.1-20 μmol/L). We evaluated the migration and invasion in Caco-2 and HT-29 CRC cell lines, impact on barrier integrity and metabolism of mycotoxins in an intestinal in vitro co-culture model comprising Caco-2/HT-29-MTX-E12/CCD-18Co. Our results showed that high glucose concentration indeed modulates the response of CRC cells to mycotoxins, affecting their metabolism and altering the cellular sensitivity towards them. This observation shed new light on the importance of the involvement of different physiological and pathological conditions when considering the toxicity of mycotoxins.

Zusammenfassung Die Senatskommission zur gesundheitlichen Bewertung von Lebensmitteln (SKLM) der Deutschen Forschungsgemeinschaft (DFG) existiert seit den frühen 1950er‐Jahren und hat sich über Jahrzehnte kontinuierlich weiterentwickelt. Sie arbeitet in Themenauswahl und Prioritätensetzung in wissenschaftlicher Freiheit und ist im Bereich der Lebensmittelsicherheit in Deutschland eine verlässliche, mandats‐unabhängige wissenschaftsgetriebene Instanz. Das Ziel der Kommission ist es, offene wissenschaftliche Fragen zur Lebensmittelsicherheit systematisch zu analysieren, Bewertungskriterien weiterzuentwickeln und wissenschaftlich fundierte Empfehlungen für gesundheitliche Bewertungen abzuleiten. Dies schließt neue Lebensmittel, Zusatzstoffe, Prozesskontaminanten und Auswirkungen der Behandlungsverfahren auf Lebensmittel ein. Kontroverse Diskussionen über mögliche Risiken durch Lebensmittel begleiten die Arbeit der Lebensmittel‐ und Ernährungswissenschaften seit Jahrzehnten und kommen häufig dadurch zustande, dass für die Bewertung erforderliches Grundlagenwissen fehlt. Die Vorgehensweise der SKLM unterscheidet sich von jener offizieller öffentlicher Institutionen wie dem Bundesinstitut für Risikobewertung (BfR) oder der Europäischen Lebensmittelbehörde EFSA (European Food Safety Authority), da sie den Schwerpunkt auf das proaktive Schließen von Erkenntnislücken sowie die Entwicklung neuer Bewertungskonzepte und Sicherheitsstrategien legt. Ein Beispiel für den Mehrwert ist etwa die Diskussion der unterschiedlichen Richtwerte, die von Regulierungsbehörden zu Bisphenol A (BPA) herausgegeben wurden, und die Nutzung dieser Diskussion, um ein zukunftsorientiertes Konzept zur Verbesserung des Prozesses zu entwickeln. Die neu gegründete „International Commission on Food Safety“ (ICFS) wird solche Aufgaben künftig von der SKLM übernehmen.

The occurring food contaminants alternariol monomethyl ether (AME), tenuazonic acid (TeA) and altertoxin II (ATX-II) are recognized as emerging Alternaria mycotoxins, yet data gaps remain regarding their toxicokinetic characteristics. The hepatic metabolism of these three substances was investigated in primary rat (PRH) and human (PHH) hepatocytes by monitoring parent compound depletion and, where applicable, metabolite formation. AME was initially evaluated at 1 µM over 4 h and subsequently investigated in concentration-dependent experiments (0.75, 1.5, 3, and 8 µM). TeA was assessed at 5 µM over 4 h, whereas ATX-II was evaluated at 1.11 µM over 4 h and at 0.22 µM over 30 min. For AME, time-dependent depletion was further investigated in PRHs at two hepatocyte densities (0.25 and 0.5 × 10⁶ cells/mL). In PRHs, AME metabolism followed Michaelis–Menten kinetics (Vmax = 150.9 pmol·min⁻¹·10⁻⁶ cells, Km = 1.18 µM), whereas no reliable kinetic model could be established for PHHs. In contrast, TeA exhibited high metabolic stability, with only 9-10% depletion after 4 h, indicating negligible hepatic clearance in both species. ATX-II was also rapidly depleted and became undetectable within 30 min, accompanied by formation of altertoxin I (ATX-I), which was more pronounced in PHHs than in PRHs. Substrate depletion revealed interspecies differences in hepatic clearance capacity and stability. Furthermore, tentative metabolite analysis indicated that AME undergoes phase I and phase II biotransformation. Overall, these findings provide comparative insights into PRH and PHH systems, offering a foundation for future studies on their toxicological relevance and impact on human health.

Human exposure to mycotoxins occurs predominantly through the ingestion of contaminated food. Despite this, potential interactions between mycotoxins and food matrix, as well as the influence of digestive processes, remain largely unexplored. In the present study, freshly prepared tomato purée was spiked with a complex Alternaria mycotoxin extract (1.2 mg/g) and subjected to a standardized in vitro digestion procedure. Changes in mycotoxin bioaccessibility throughout the consecutive digestive phases were comprehensively monitored by LC-MS/MS analysis. The COMET assay and AMES test were conducted to investigate differences in genotoxicity and mutagenicity between the undigested and digested extract, respectively. In vitro digestion altered the concentrations of most mycotoxins present in the bioaccessible fraction, thereby partially reducing the extract's genotoxic and mutagenic potential. These results emphasize the need to account for digestive processes and food matrices when assessing mycotoxin toxicity and provide a robust framework for more realistic future risk assessment strategies.

Our environment, especially our diet, exposes us to numerous bioactive compounds that have the potential to interact with anticancer therapies. Although understanding these exposome-drug interactions is crucial for optimizing anticancer therapy, limited knowledge exists on the interference of contaminants, especially mycotoxins, with anticancer drugs. One of the most prevalent mycotoxins in food and feed is zearalenone (ZEN), which demonstrated estrogenic activity that is even stronger in its metabolite α-zearalenol (α-ZEL). Consequently, in this study, we investigated whether these mycoestrogens affect estrogen-dependent breast cancer therapy with doxorubicin, a standard treatment option. The effects of mycoestrogens on doxorubicin were investigated in MCF7 estrogen receptor-positive breast cancer cells. Cell viability assays revealed that both ZEN (between 1 and 10 µM) and α-ZEL (>0.01 µM) reduced the anticancer activity of doxorubicin. This decrease in doxorubicin activity by α-ZEL, and to a lesser degree ZEN, was due to a decrease in cell death (up to 30 % reduction of annexin V signal by both, 50 % more protein expression of the apoptosis-related proteins B-cell lymphoma 2 and 25 % less Bcl-2-associated X protein by 0.1 µM α-ZEL) and stimulation of the cell cycle progression (up to 6 % more cells in S phase and 5 % less in G2/M phase, and changes in protein expression of cell cycle-related proteins cyclin D1, p16, p21 and p53 by α-ZEL). Application of an estrogen receptor α inhibitor linked the ability of α-ZEL to interfere with cell viability and apoptosis to its activation of the receptor. Consequently, contamination of our diet with mycoestrogens might not only impact cancer growth but also impair the effectiveness of anticancer therapy in patients with estrogen-dependent cancer, warranting greater attention in its management.

Lipophilic coffee compounds are present in coffee beverages and may also be found in coffee by-products, whose underutilized fractions contain molecules such as RN-alkanoyl-5-hydroxytryptamines (Cn-5HTs) and diterpenes. Although members of these groups show anti-inflammatory potential, their combined effects remain unexplored. In this study, Cn-5HTs, cafestol, kahweol, and caffeine were tested individually and in mixtures in THP-1 LuciaTM monocytes using an NF-kappa B reporter gene assay to evaluate additive or synergistic interactions. The most active combinations were further assessed in HCEC-1CT intestinal epithelial cells for effects on TNF-alpha, IL-6, IL-8, and IL10 expression and secretion. C18-5HT, C20-5HT, cafestol, and kahweol reduced NF-kappa B activation at non-cytotoxic concentrations, while caffeine and longer-chain amides were inactive. C18-5HT and kahweol were the most potent, and their co-treatment (3 & micro;M + 40 & micro;M) produced the strongest inhibition, suggesting synergy. In HCEC1CT cells, treatments lowered pro-inflammatory cytokine mRNA and protein levels, supporting the bioactivity of coffee lipophilic compounds.

Multiple mycotoxins in feed threaten animal health and food safety, demanding sustainable mitigation strategies. This study evaluated acid-modified mangosteen peel (AMP), an agricultural by-product, as a potential multi-mycotoxin adsorbent. Physicochemical characterization using scanning electron microscopy (SEM), Brunauer-Emmett-Teller (BET) surface area analysis, and Fourier transform infrared spectroscopy (FTIR) analyses demonstrated that acid modification increased surface area (1.9 to 9.03 m2/g), pore volume (0.005 to 0.027 cm3/g), and surface negativity, indicating enhanced adsorption properties. In vitro binding experiments assessed adsorption of aflatoxin B1 (AFB1), zearalenone (ZEA), ochratoxin A (OTA), T-2 toxin, deoxynivalenol (DON) and fumonisin B1 (FB1) under different pH conditions. AMP exhibited high adsorption efficiencies for AFB1, ZEA, OTA, and T-2 toxin, particularly at pH 3, whereas DON and FB1 showed limited binding. Adsorption behavior was dose-dependent and best described by Langmuir and Freundlich isotherm models. Simulated gastrointestinal digestion indicated stable binding of AFB1 and ZEA under gastric conditions, with partial release of some toxins at neutral pH. Cytotoxicity assessment in porcine intestinal epithelial cells (IPEC J2) showed no apparent cytotoxic effects at 0.25-1 mg/mL. Therefore, AMP demonstrated improved multi-mycotoxin adsorption compared to the untreated material and showed no apparent cytotoxic effects in vitro within the tested concentration range, indicating its potential as a promising feed additive candidate.

Peptide drugs have revolutionized modern medicine owing to their high potency, selectivity, and excellent tolerability. However, oral delivery remains limited, and most peptide drugs are administered parenterally due to their inherent instability to proteolytic digestion and poor ability to cross gastrointestinal barriers, which hinders efficient absorption into the bloodstream. This study presents a multifunctional oral delivery system based on mesoporous silica nanoparticles (MSN) customized for insulin administration. Insulin-loaded MSN were co-formulated with succinylated β-lactoglobulin to produce pH-responsive tablets that limited premature gastric release (≤13% after 2 h at pH 1.2) and protected insulin from enzymatic degradation, while enabling controlled intestinal release (up to 88%-98% at pH 7.4). Surface functionalization with polyethylene glycol and phosphonate moieties improved colloidal stability and increased insulin solubility by ∼2.5-fold. The interaction of phosphonated MSN with intestinal epithelial cells further induced transient reorganization of tight junction proteins, enhancing paracellular insulin transport (26% after 24 h, compared with 13% for non-confined insulin). Delivered insulin retained bioactivity, as demonstrated by activation of insulin-responsive signaling pathways in vitro and reduced blood glucose levels in hyperglycemic mice. These results highlight MSN as a promising platform for oral peptide delivery with improved efficacy and patient compliance.

Abstract Tenuazonic acid (TeA) is an emerging Alternaria mycotoxin frequently detected in food and feed commodities, raising concerns about its toxicological relevance. Chronic oral exposure to TeA has been reported to induce dysplastic alterations in the esophageal mucosa of mice, while human biomonitoring data indicate an association between TeA exposure and esophageal cancer, although a causal relationship has not yet been established. At a mechanistic level, the effects of TeA in esophageal cells remain poorly characterized. Therefore, this study investigated the impact of TeA on cytotoxicity, oxidative stress, DNA damage, mitochondrial homeostasis, cell-cycle distribution and transcriptomic stress responses in human esophageal KYSE-510 cells. TeA induced a concentration-dependent reduction in metabolic activity and total protein content after 24 h exposure to 0.1-100 µM. Significant cytotoxicity was measured starting from 20 µM. At sub-cytotoxic concentrations, TeA triggered rapid ROS formation within 5-30 min exposure and induced formamidopyrimidine-DNA glycosylase (FPG) sensitive DNA damage after 1 h exposure (5-7.5 µM), indicating oxidative DNA lesions. In addition, TeA altered mitochondrial morphology after 4 h exposure at 7.5 µM, manifested by shrinkage of the mitochondrial network area and perinuclear redistribution, while mitochondrial respiration showed only a non-significant tendency towards reduced respiratory capacity. RNA sequencing after 6 h exposure to 10 µM TeA revealed oxidative stress-associated transcriptional changes, impaired antioxidant and stress-adaptive responses, and p53-associated stress signaling. Furthermore, TeA induced significant G₂/M phase accumulation after 24 h exposure to 1-10 µM. Highlights TeA triggers rapid oxidative stress in human esophageal KYSE-510 cells Sub-cytotoxic TeA induces FPG-sensitive oxidative DNA lesions TeA disrupts mitochondrial morphology before overt cytotoxicity occurs RNA-seq reveals impaired antioxidant defense and p53-linked stress signaling TeA promotes G₂/M phase accumulation and concentration-dependent cytotoxicity Graphical abstract

Enniatins (ENNs) and beauvericin (BEA) are emerging Fusarium mycotoxins frequently detected in cereals and processed foods, raising concerns about their toxic potential. These cyclic hexadepsipeptides exert cytotoxic effects by disrupting ionic homeostasis and accumulating in lipid membranes. Although ENNs and BEA modulate pathways such as NF-κB and MAPK, their direct immunomodulatory effects remain poorly characterized. This study investigates their impact on inflammation, cytokine regulation, and metabolic activity in human immune (THP-1 Lucia™ monocytes) and intestinal (HCEC-1CT, Caco-2) cell models, including effects of phase I metabolites generated by S9 mix preincubation. In THP-1 Lucia™ monocytes, all tested ENNs (2.5-5 µM) and BEA (5 µM) suppressed lipopolysaccharide-induced NF-κB activity only at cytotoxic concentrations, suggesting that immunosuppressive effects are secondary to cell damage. Pre-treatment with rat liver S9 mix mitigated cytotoxicity and attenuated NF-κB suppression. In intestinal cells, ENNs and BEA reduced TNF-α transcription in non-tumorigenic HCEC-1CT but not in tumorigenic Caco-2 cells, indicating cell-type-specific responses. These findings emphasize the context-dependent immunotoxicity of ENNs and BEA and underscore the need to distinguish between direct immunomodulation and secondary effects of reduced cell viability. While broader immune disruption appears limited to toxic concentrations, TNF-α suppression in non-tumorigenic intestinal cells may pose a health concern.

The health benefits of soy and soy derived foods are often attributed to their isoflavone (ISFs) content, known to include phytoestrogens. However, EFSA has concluded that there is insufficient scientific evidence to establish a causal relationship between ISFs consumption and the proposed health claims. Although phase-II metabolites of ISFs are often considered as detoxified products with reduced estrogenic effects, recent studies indicate that ISFs metabolites may contribute and even enhance the biological activity of ISFs. However, current knowledge on phase-II metabolites of ISFs is limited, even though representing the major ISFs metabolites circulating in the bloodstreams in humans. This study investigates the role of phase-II metabolism in the biological activity of ISFs, focusing on estrogenicity and cytotoxicity in estrogen-sensitive Ishikawa cells (expressing estrogen receptors (ER) α and β). ISFs concentrations from 0.001 to 10 µM, with 1:10 dilution steps, were tested. Cleavage to parent compounds during incubation was also evaluated using high-performance liquid chromatography tandem mass spectrometry. The results showed that phase-II metabolites, particularly sulfates, can still exhibit estrogenic activity, challenging the general assumption that phase-II metabolism always leads to the inactivation of ISFs. In particular, at 10 µM daidzein-4'-sulfate exhibited a comparable estrogenic activity to daidzein at the same concentration, and genistein-7-sulfate exhibited estrogenic potential at higher concentrations. Molecular dynamics simulations revealed that genistein and its conjugates can stably interact with ERβ. This study highlights the importance of phase-II metabolism for ISFs activity and suggests that the sulfates formed should be examined more closely in further investigations.

Alternaria species produce structurally diverse mycotoxins that frequently contaminate food and are discussed in association with adverse health effects. However, systematic knowledge of their immunosuppressive potential is limited. This study evaluated seven Alternaria mycotoxins-alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), alterperylenol (ALTP), altertoxin I (ATX-I), altersetin (AST), and tentoxin (TEN)-in immune and intestinal cells. NF-κB reporter gene assays were performed in THP-1 monocytes and phorbol 12-myristate 13-acetate-differentiated macrophages (0.01-25 µM), while cytokine transcription and secretion (IL-6, IL-8, IL-10, TNF-α) were analyzed in Caco-2 and HCEC-1CT cells (0.1–30 µM) applying qRT-PCR and ELISA. All mycotoxins suppressed NF-κB activation in immune cells, though with differing potency. AOH, ALTP, and ATX-I were active at concentrations ≥ 1 µM in both models. AME and AST were more potent in macrophages (≥ 0.01 µM) than monocytes (≥ 5 µM). TEN suppressed the pathway at ≥ 0.1 µM in macrophages and ≥ 5 µM in monocytes, while ALT at ≥ 5 µM in both cell types. In intestinal models, AOH, AME, ALTP, ATX-I, and TEN modulated cytokine transcription, with stronger responses in non-tumorigenic HCEC-1CT cells. ALTP emerged as the most potent immunosuppressive compound, reducing IL-6, IL-8, and/or TNF-α secretion at concentrations ≥ 0.1 µM. AOH showed dual activity, suppressing IL-6/IL-8 at 30 µM, while modulating TNF-α in a cell-type-dependent manner (increased secretion in Caco-2, decreased in HCEC-1CT). This comparative study reveals immunosuppressive profiles of Alternaria mycotoxins in immune and epithelial cells, highlighting their relevance as food contaminants capable of modulating both intestinal and systemic immune responses.

The foodborne mycotoxins alternariol (AOH) and alternariol monomethyl ether (AME) have been associated with several adverse effects, including cytotoxicity, genotoxicity, endocrine disruption, and immunomodulation. As these endpoints are typically observed in vitro at micromolar concentrations, the question arises whether such levels are attainable in exposed humans. To address this data gap in chemical risk assessment, a physiologically based kinetic (PBK) model was developed to predict internal exposure doses to AOH and AME in humans. As input parameters, kinetic constants for hepatic glucuronidation were obtained in vitro by incubating Sprague Dawley rat and human liver S9 fractions with 0.5–50 µM AOH and 0.5–20 µM AME, demonstrating rapid biotransformation in both species. Intestinal absorption of AME and physicochemical parameters were estimated using quantitative structure–activity relationship (QSAR) models. Sensitivity analysis identified parameters describing hepatic glucuronidation and gastrointestinal uptake as among the most influential, confirming the importance of their reliable estimation. The PBK model was evaluated against available rodent toxicokinetic data and subsequently extrapolated to humans. Ultimately, the currently available exposure estimates published by EFSA in 2016 were applied to predict target tissue concentrations, which were compared to points of departure (PoDs) for relevant toxicological endpoints. Even in the most susceptible group of male toddlers, predicted internal concentrations (10⁻4 µM range) were approximately four orders of magnitude below the respective PoDs. Consequently, under the applied exposure assumptions and considering the compounds as isolated chemicals, AOH and AME are not expected to reach systemic or tissue concentrations associated with the investigated effects.

Humans and animals are frequently exposed to dietary compounds that can adversely affect intestinal health. The Fusarium mycotoxin deoxynivalenol (DON) is known to compromise epithelial barrier integrity. DON frequently co-occurs in food and feed with the mycoestrogen zearalenone (ZEN) and soy-derived isoflavones (ISFs) such as genistein (GEN) and daidzein. Therefore, this study investigates the effects of ZEN (10 nM), its metabolites hydrolyzed ZEN (HZEN) and decarboxylated hydrolyzed ZEN (DHZEN) (each 10 nM), and ISFs (GEN and equol (EQ), 10 µM), alone and in combination with DON (10 µM), on intestinal barrier function using the porcine epithelial cell line IPEC-J2. Barrier integrity was assessed via transepithelial electrical resistance (TEER) and paracellular permeability (lucifer yellow). DON (10 µM) significantly reduced barrier integrity, confirming its disruptive effect. Conversely, treatment with ZEN, HZEN, DHZEN, GEN, and EQ did not impair barrier function. Co-exposure of DON with ZEN or HZEN yielded TEER values similar to those observed with DON alone. In contrast, the DHZEN + DON combination resulted in higher TEER values compared to DON alone, suggesting a mitigating effect under the experimental conditions. Quantification of GEN by LC-MS/MS showed consistent levels across treatments, except for reduced basolateral GEN during co-incubation with DHZEN, suggesting altered transport or metabolism. These findings highlight the ability of DHZEN to mitigate the intestinal barrier disruption induced by DON and underscores the complex interaction of maintaining epithelial barrier integrities.

Intestinal barrier integrity is of paramount importance for our health. Several food constituents and contaminants have been described for positive or detrimental effects, however, a correlation between gut associated disorders and xenobiotics exposures remains challenging and modeling options limited. This study presents a NAMs-based hybrid in vitro/in silico workflow to investigate, and eventually prioritize, foodborne xenobiotics for their effect on gut barrier homeostasis. Model compounds comprised endocrine-active chemicals, such as plastic components (bisphenol A, bisphenol F), phytoestrogens (chrysin, daidzein, genistein), mycotoxins (alternariol, zearalenone), and natural/synthetic steroids (17-β-estradiol, cholesterol, levonorgestrel). Plasticizer diisodecyl phthalate was included as foodborne, non-endocrine active substance. The Caco-2/HT29-MTX-E12 intestinal epithelial co-culture was used to design a workflow capable of grasping discrete alterations in tight junction organization. The protein claudin-4 was taken as reference to study the structural organization along cell-cell contacts. Complementarily, docking and molecular dynamics simulations were applied to explore ligand-claudin-4 interactions, providing a pipeline which is potentially suitable to screen larger compounds libraries. Finally, proof-of-principle experiments including repeated exposures to reference compound 17-β-estradiol and plasticizer diisodecyl phthalate, supported the plausibility of a functional impairment of claudin-4. Both compounds significantly increased epithelial permeability without altering cell viability, indicating selective modulation of mechanical barrier properties. Together, this work outlines a mechanistic toolbox for the study of the potential of orally derived xenobiotics in modulating intestinal homeostasis.

Abstract The ubiquitously occurring food contaminants altenuene (ALT) and tentoxin (TEN) are recognized as emerging Alternaria mycotoxins, yet substantial data gaps remain when it comes to their toxicological behavior and toxicokinetic characteristics. This study aimed to compare and generate quantitative data on their hepatic metabolism and to obtain semi-quantitative insights into their metabolite profiles. To this end, primary rat and human hepatocytes were incubated with 10 µM ALT or TEN over multiple time points up to 4 h. Both substrate depletion and metabolite identification revealed pronounced interspecies differences. The extent of ALT metabolism was significant, with an 88% and 57% decrease in rat and human hepatocytes after 4 h, respectively. In contrast, TEN showed extensive biotransformation in rats (67%) but only modest turnover in humans (27%) over the same period. Hepatocellular clearances were consistently higher for ALT than TEN, with hepatic extraction ratios indicating intermediate extraction for ALT and low extraction for TEN. High-resolution mass spectrometry combined with targeted analysis of selected metabolites annotated phase II conjugation as the predominant metabolic pathway for ALT and phase I oxidative metabolism for TEN, including mono- and double-metabolized species for the latter. Overall, these results provide a comprehensive characterization of ALT- and TEN-metabolism in hepatocytes, offering a foundation for future studies on their toxicological relevance and impact on human health.

Schizophyllum commune mycelia obtained from submerged fermentation were investigated as a sustainable source of bioactive ingredients using an integrated workflow involving green extraction, proteomics, in silico peptide-motif prediction, and Caco-2 cellular assays. Hot-water extraction (HWE) and ultrasound-assisted extraction (UAE) were compared at different biomass loads and extraction times. Protein solubilization increased strongly with biomass load regardless of extraction method, whereas polysaccharide recovery depended mainly on extraction time. In crude extracts, UAE gave higher phenolic content than HWE. Shotgun proteomics identified 312 high-confidence proteins, from which selected proteins were used as candidate precursor sequences for antioxidant-peptide motif prediction. The hydrolysate, which contained peptides together with polysaccharides and phenolic compounds, maintained >90% Caco-2 viability and significantly reduced H2O2-induced intracellular reactive oxygen species in a dose-dependent manner. These findings provide preliminary support for the further development of the hydrolysate as a multicomponent antioxidant ingredient for functional food applications.