Search Results (10,052)

The globalization of the food supply chain presents complex challenges for safety assurance within a highly fragmented regulatory landscape. This review synthesizes the frameworks of eight influential jurisdictions—including the European Union (EU), the United States, China, and Codex Alimentarius—to evaluate how legal mandates function as regulatory drivers that guide the evolution of analytical chemistry. By examining legislation on Maximum Residue Limits (MRLs), positive list systems, and method validation guidelines (e.g., SANTE), we demonstrate that strict preventive controls have established chromatography coupled with tandem mass spectrometry (LC/GC-MS/MS) as the universal standard for multi-residue screening. We show that global regulatory fragmentation is not merely an administrative artifact, but is rooted in divergent toxicological philosophies and localized dietary exposure models. This regulatory heterogeneity requires analytical laboratories to adopt a posture of “defensive technological redundancy,” forcing them to continuously optimize targeted methods against the strictest global default limits (e.g., 0.01 mg/kg). We establish that this continuous methodological escalation for ultra-trace quantification has reached practical and operational limits. Consequently, we conclude that the future of food safety testing must transition from static target-list compliance toward adaptable, non-targeted chemical profiling using High-Resolution Mass Spectrometry (HRMS), enabling laboratories to proactively address emerging contaminants, food fraud, and the complexities of modern food matrices. Full article

The identification and characterization of air pollutants in metropolitan environments are of paramount global concern due to their significant implications for air quality and public health. This study investigates the chemical composition of fine particulate matter (PM_2.5) at two strategically selected urban sites in Seoul, South Korea, during 2020: Gwanghwamun Plaza, representing a high-density central location, and Bokjeong Station, situated in the metropolitan periphery. A key aspect of this research is the detection of terephthalic acid (TPA)—a distinct marker of polyethylene terephthalate (PET) combustion—using high-resolution liquid chromatography–time-of-flight tandem mass spectrometry (LC-ToF-MS/MS). Results from the simultaneous measurement campaign demonstrate that nighttime conditions strongly influence PM_2.5 at both sites, with increases observed not only in absolute concentrations (levoglucosan, TPA, As, CO, and NH₃) but also in OC-normalized ratios (levoglucosan/OC and TPA/OC). The consistent nighttime enhancement of these ratios suggests that the observed increases cannot be explained solely by reduced planetary boundary layer height but instead indicate relatively stronger emission contributions. These increases are likely influenced by waste incineration activities, wherein PET-based plastics and wood materials are combusted. Furthermore, assessment of the dithiothreitol assay-derived oxidative potential (DTT-OP) underscores the heightened oxidative stress associated with these emissions, posing substantial health risks. Full article

Aflatoxin B1 (AFB1) is a potent mycotoxin threatening food and feed safety. Here, we report the identification and characterization of a Bacillus safensis-derived multicopper oxidase (BsaMCO) capable of efficient AFB1 detoxification. Recombinant BsaMCO exhibited robust in vitro activity, achieving >78% degradation of AFB1 under 24 h incubation at 37 °C. Optimization experiments revealed that enzyme concentration, pH, temperature, metal ions, and electron acceptors significantly influenced degradation efficiency, defining an operational window suitable for practical applications. LC–MS profiling suggested the presence of transformation products tentatively consistent with oxidative demethylation to aflatoxin P1 (AFP1) and with the formation of AFG2a-like products through subsequent hydration- and oxidation-related transformations. Molecular docking and 100 ns all-atom molecular dynamics (MD) simulations demonstrated stable binding of AFB1 in the T1 copper pocket. Van der Waals and electrostatic interactions, together with a persistent hydrogen bond at Gly323, facilitated single-electron transfer through the intramolecular T2/T3 copper cluster. Principal component and Gibbs free energy analyses confirmed a low-energy, stable conformational ensemble. HepG2 cell assays indicated that BsaMCO-degraded products substantially reduced cytotoxicity and apoptosis compared with native AFB1. Simulated feed experiments further validated enzymatic AFB1 degradation, with approximately 53% reduction after 24 h. Collectively, these findings establish BsaMCO as a safe and effective biocatalyst for AFB1 detoxification, providing mechanistic, structural, and cellular evidence supporting its application in food and feed safety. Full article

Recent advancements in neuroproteomics have enabled detailed analysis of protein expression in the human brain, yet resolving synaptic dysfunction—a central feature of many neurological and psychiatric disorders—requires careful methodological consideration. Leveraging the high sensitivity of modern liquid chromatography-tandem mass spectrometry (LC-MS/MS), we evaluated the utility of whole-tissue lysates versus enriched synaptosome preparations for detecting synaptic protein signatures. First, we optimized and standardized a sample preparation protocol for frozen human gray matter (GM) by refining the suspension trapping (sTRAP) digestion method using thin human tissue sections. We accomplished low technical variation by minimizing sample handling and achieved a highly reproducible sample preparation workflow by rigorously applying standardization and randomization across dissection, processing, and LC-MS/MS runs. Second, comparative LC-MS/MS analysis showed that while whole-tissue lysates provide a high-throughput survey of the synaptic proteome, synaptosome isolation is required to investigate synapse-specific proteins to detect alterations at the terminal that are obscured in the soma. Because these methods offer distinct but synergistic levels of information, we recommend a tiered neuroproteomics strategy. This approach utilizes whole-tissue lysates for broad disease-associated screening and consistent quantification in large cohorts, followed by targeted synaptosome proteomics to provide a unique window of insight into synaptic composition and stability. This integrated workflow respects the biological necessity of spatial resolution while maintaining the reproducibility required for robust human brain proteomics. Furthermore, initial tissue-level analysis provides the necessary context to correctly interpret synaptosome data in cases of global synapse loss or gain. Full article

Dezhou donkey is a premium indigenous Chinese livestock breed with high economic value for meat, hide and medicinal uses, and growth rate is a core trait determining farming profitability. However, the molecular and metabolic mechanisms underlying divergent growth rates in this breed have not been fully characterized, with no integrated transcriptomic and metabolomic studies reported. Here, 12 age-matched healthy male Dezhou donkeys were assigned to faster-growing (n = 6) and slower-growing (n = 6) groups by average daily gain, followed by plasma transcriptome sequencing and untargeted LC-MS/MS metabolomics. We identified 480 differentially expressed genes, with the slower-growing group enriching in immune/inflammatory/apoptotic pathways, and the faster-growing group in energy metabolism and transmembrane transport. Lipids and lipid-like molecules represented the largest proportion (44.9%) of the differential metabolites; the slower-growing group was enriched in lipid peroxidation and pro-inflammatory mediators, while the faster-growing group was enriched in unsaturated fatty acids and antioxidants. Integrated analysis revealed core pathways (cAMP signaling, arachidonic acid/unsaturated fatty acid biosynthesis) and key candidate genes/metabolites. Our findings clarify that excessive lipid peroxidation and inflammatory imbalance restrict growth, while efficient energy metabolism promotes faster growth, providing theoretical support for genetic improvement and precision nutrition of Dezhou donkeys. Full article

The European Regulation (UE) 432/2012 includes a specific health claim for olive-oil-associated with its phenolic content, which is based on its protective role against lipid oxidation in the blood. To make use of the health claim, olive oil must have a minimum concentration in phenolic compounds of 250 mg/kg. Reviewing the health claim, the phenolic compounds referred to are the secoiridoid derivatives of hydroxytyrosol and tyrosol. A method based on absolute quantification of phenolic compounds in olive oil is proposed for the recognition of the health claim. The method involves liquid–liquid extraction with a 1:8 (v/v) oil:extract ratio to avoid saturation of the extract in oils with a higher phenolic content and its subsequent determination through LC–MS/MS in multiple reaction monitoring (MRM) mode, the gold standard technique in many application fields because of its analytical features. The optimized method was applied to a set of 100 extra virgin olive oils (EVOOs), and the results obtained were compared with the classic Folin–Ciocalteu method. The comparison between the two methods showed that the classic method is a non-selective method that can be affected by many interferences and that the Folin method underestimates the real phenolic content. Full article

The non-thermal plasma (NTP) process is a promising advanced oxidation process (AOP) for removing non-biodegradable organics from wastewater, owing to the efficient formation of reactive chemicals. Despite its effective oxidizing capability, the decomposition mechanism of organic pollutants is not well understood. This study evaluates NTP for two representative sulfonamides (SMZ and STZ) and reports on (i) time-resolved removal to the method detection limit, (ii) transformation mapping using LC-ESI/MS/MS, which confirmed previously proposed hydroxylation and bond-cleavage pathways and further identified additional hydroxylated intermediates formed on the thiazole and benzene rings under NTP conditions, and (iii) energy evaluation through energy per order (EEO) within a single, reproducible operating window. The EEO values for SMZ and STZ degradation via NTP were calculated at 22.4 and 7.5 kWh/m³/order, respectively. These values are up to 37- and 118-fold lower than those reported for comparable AOPs, quantitatively confirming that the proposed NTP process achieves superior energy efficiency for sulfonamide degradation. Degradation is primarily attributed to reactive oxygen species (ROS) generated by plasma, which initiate the breakdown of the antibiotic structure. Overall, this study demonstrates that NTP is a highly effective AOP for driving the rapid primary degradation and intermediate structural transformation of recalcitrant sulfonamide antibiotics. Full article

Background/Objectives: MYC-driven tumors exhibit significant glutamine addiction, but the metabolic adaptation mechanisms enabling their survival under glutamine deprivation remain incompletely understood. Malic enzymes catalyze the oxidative decarboxylation of malate to pyruvate while generating NADPH, linking central carbon metabolism to redox homeostasis. This study investigates whether and how ME1 and ME2 mediate cell adaptation to glutamine starvation and explores their functional division in relation to p53 status. Methods: Using MYC-amplified, p53-mutant (G266E) SF188 glioblastoma cells, we performed siRNA-mediated knockdown, overexpression, and rescue experiments. Cell survival was assessed by trypan blue exclusion and Annexin V/PI staining. ROS levels and NADP⁺/NADPH ratios were measured by DCFH-DA fluorescence and enzymatic assays. Metabolite tracing was conducted using [U-¹³C₅] glutamine followed by LC-MS. Key findings were validated in additional cell lines including HCT116, U2OS and MDA-MB-231. Results: ME1 and ME2 promote SF188 cell survival under glutamine deprivation, an effect that depends on their catalytic activity but is independent of TCA cycle anaplerosis. ME1 maintains redox balance by generating NADPH, and antioxidant treatment rescues the survival defect caused by ME1 knockdown. In contrast, ME2 does not contribute to redox regulation but stabilizes mutant p53 (G266E) via proteasome inhibition. Both of these pro-survival functions are attenuated upon MYC knockdown, suggesting a dependency on MYC expression. Across all cell lines tested, ME1 and ME2 also promote survival through redox maintenance, although the isoform responsible for antioxidant function differs. Conclusions: ME1 and ME2 support metabolic adaptation to glutamine starvation through distinct, isoform-specific mechanisms that depend on MYC expression and p53 mutation status. These findings suggest malic enzymes as potential therapeutic targets in MYC-driven, p53-mutant tumors. Full article

Background: Sensors and mass spectrometry (MS) are frequently used in combination for food safety and quality assessment, yet their functional integration lacks a formal methodological framework. This review categorizes the synergies between these technologies into distinct Relational Connections. Methodology: Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, 155 original research articles published between 2015 and 2025 were systematically analyzed. Records were identified via the Scopus database within the food science domain. Experimental meta-data, including extraction protocols, instrumental configurations (ionization source, mass analyzer, cost tier), and chemometric strategies, were extracted to identify core methodological patterns. Statistical associations were quantified using chi-squared tests with Cramer’s V effect sizes. Results: Five Relational Connections were identified: (1) MS as reference for sensor validation (25.2%); (2) MS-sensor correlative analysis (10.3%); (3) MS quantifying data to train predictive sensor models (6.5%); (4) MS identifying targets for sensor detection (7.1%); and (5) MS enabling sensor classification models (51.0%). Technology pairing is governed by a three-level hierarchy: analyte polarity determines the ionization source (V = 0.69), required precision determines the mass analyzer (V = 0.64), and cost/availability constraints shape the practical integration strategy. Gas Chromatography (GC)-MS is predominantly coupled with Electronic Noses for volatile profiling (86% of classification studies), while Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) pairs with biosensors for contaminant analysis (74% of reference validation studies). Systematic analysis of the full pairing matrix reveals that 75% of theoretically possible MS-sensor combinations remain unexplored or underrepresented, identifying both technical boundaries and innovation frontiers. Discussion: The findings clarify the strategic logic behind technology pairings, demonstrating that MS provides the quantitative molecular data required for sensor training. The hierarchical decision framework and identification of underexplored pairings provide an evidence-based guide for designing future integrated food analysis systems. Full article

Cajanus cajan (L.) Huth (Fabaceae) is a food legume of considerable nutritional and functional significance. This study examined the comparative effects of salt stress on seed germination, hydroponic growth, and phytochemical accumulation across two developmental stages: 10-day-old germinated seeds and 45-day-old hydroponically grown plants, using NaCl solutions at concentrations of 0, 25, 50, 75, 100, and 150 mM. Both germination rate and growth were greatest at 0–25 mM NaCl, with performance declining at higher concentrations. LC–MS/MS analysis of free amino acids in 10-day-germinated seeds revealed a salt-induced metabolic shift. Proline, leucine, and phenylalanine were the dominant free amino acids and increased progressively with rising NaCl concentrations. Phytochemical profiling by HPLC identified gallic acid, catechin, and genistin as the major compounds, with increased levels under salinity stress. Germinated seeds at 150 mM NaCl, germinated seeds exhibited the highest phytochemical accumulation, with total phenolic content (TPC), total flavonoid content (TFC), and DPPH activity reaching 18.192 ± 0.020 mg GAE/g extract, 8.519 ± 0.026 mg QE/g extract, and 11.623 ± 0.284 mg AAE/g extract, respectively. Phytochemical responses in 45-day hydroponic plants varied by tissue type. Leaves exhibited declining TPC and TFC with increasing NaCl (from 29 to 16 mg GAE/g and 41 mg QE/g extract), while stems showed the opposite trend, reaching 18 mg GAE/g and 21 mg QE/g extract at 50 mM. Root tissues maintained comparatively low phytochemical levels throughout. Notably, DPPH scavenging capacity increased across all tissues under salt stress, with peak values of 12–13 µg AAE/g extract recorded at 50 mM NaCl. These results indicate that salt stress exerts stage- and organ-dependent effects on phytochemical accumulation in C. cajan. High salinity during germination stimulates bioactive compound production, whereas moderate salinity appears to be the threshold at which antioxidant capacity is maximized in hydroponic systems. These observations point to the practical utility of controlled salt elicitation as a strategy for enriching pigeon pea with health-promoting phytochemicals, reinforcing its potential as a functional food crop. Full article

This work presents a comparative study of micro- and nano-scale fillers in liquid composite molding processes, focusing on how particle size and morphology affect resin rheology, flow behavior, and filler filtration within fiber preforms. Glass microspheres and organo-modified montmorillonite were dispersed in epoxy resin and injected through glass-mat preforms at different fiber volume fractions (ranging from 0.27 to 0.47). Our study integrates rheological characterization, in situ flow-front tracking, unsaturated permeability analysis, thermogravimetric quantification of retained particles, and microstructural observations by SEM. Despite their smaller loading, nanoclay suspensions showed a markedly higher viscosity increase than microsphere systems, yet their permeability remained nearly unchanged. In contrast, microsphere-filled resins exhibited strong filtration at the flow inlet, density-driven settling near the lower tool face, and significant permeability loss. The results demonstrate that nano-fillers, although more viscous, maintain homogeneous distribution and flow continuity, whereas micro-fillers promote cake formation and local compaction. This controlled side-by-side comparison clarifies how filler size and shape govern filtration mechanisms in liquid composite molding (LCM), providing design guidelines for processing filled resin systems without compromising part quality. Full article

Siponimod, a selective sphingosine 1-phosphate (S1P) receptor modulator, represents a next-generation therapeutic drug for active secondary progressive multiple sclerosis. This study conducted in-depth forced degradation studies of siponimod in solid state subjected to acidic, alkaline, oxidative, photolytic, and thermal conditions, in compliance with ICH guidelines Q1A (R2) and Q3A (R2). An HPLC method was developed to quantify siponimod and separate its degradation products (DPs). The DPs were characterized using LC-HRMS/MS and LC-MSⁿ techniques. Moreover, the toxicological profiles of siponimod and its DPs were evaluated through the in silico tools ProTox 3.0 and ADMETlab 3.0, with molecular docking and dynamics simulations assessing their binding to the S1P1 receptor. Siponimod was stable to light but degraded under acidic, alkaline, oxidative, and thermal stress, producing five products: DP-1 (acidic), DP-2/3 (oxidative), DP-4 (hydrolytic), and DP-5 (thermal). The toxicity prediction suggested that neither siponimod nor its DPs exhibited carcinogenic or mutagenic potential, and the molecular modeling analysis revealed that DP-2 and DP-3 demonstrated favorable binding affinities, with stable dynamic profiles and thermodynamic properties that closely resembled those of siponimod. As far as we know, this is the first study on the structural elucidation of the DPs of siponimod by LC-HRMS/MS and LC-MSⁿ. Full article

Muskmelon (Cucumis melo L.) is an economically important crop that remains highly susceptible to destructive fungal diseases, including gummy stem blight, downy mildew, Fusarium wilt, and anthracnose. Although fungicides and resistant cultivars are widely used, reliance on chemical control raises concerns regarding environmental safety, food quality, and the emergence of fungicide-resistant pathogen populations. Consequently, microbial biopesticides, particularly Bacillus species, have attracted increasing attention as sustainable alternatives. In this study, muskmelon plants exhibiting leaf wilting, chlorosis, and stem yellowing were collected from Guangming Farm in Wufeng, Taichung, Taiwan, and associated pathogens were isolated from stem tissues and identified to determine the causal agent of these symptoms. In addition, the biocontrol efficacy of Bacillus subtilis strain MCLB2 against melon fruit rot, as well as its underlying mechanisms, was evaluated. Pathogenicity assays confirmed that isolate F01 was the causal agent. Based on morphological characteristics and internal transcribed spacer (ITS) sequence analysis, this isolate showed 99.8% identity to Fusarium pernambucanum URM 7559 (GenBank accession no. NR_163754), and phylogenetic analysis further placed it within the Fusarium incarnatum–equiseti species complex (FIESC). Antagonistic assays demonstrated that B. subtilis MCLB2 significantly inhibited mycelial growth and suppressed the spore germination of F. pernambucanum. In addition, culture filtrates of strain MCLB2 effectively reduced Fusarium-induced fruit rot in melon and disrupted fungal cellular respiration. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis revealed that the strain produced surfactin-family lipopeptides. In conclusion, B. subtilis MCLB2 exhibits potential as a sustainable biocontrol agent for managing Fusarium fruit rot in melon, likely through surfactin-mediated disruption of fungal cellular respiration. Full article

A homologous classification for vespid venoms is missing. This study compared Polistes dominula and Vespula spp. venoms to evaluate their homology level. P. dominula and Vespula spp. extracts, including V. germanica, V. maculifrons, V. pensylvanica, V. alascensis, and V. squamosa in equal proportions, were generated from venom sacs and were subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot using Vespula-positive sera. Bands described as allergenic were excised and sequenced through Liquid Chromatography–Mass Spectrometry tandem analysis (LC-MS/MS) to confirm their identity. Phospholipase (group 1) and hyaluronidase (group 2) enzymatic activities were measured. Group 1 and 5 3-D structures and sequence identity were analyzed in silico. The results showed that the P. dominula and Vespula spp. venom extracts exhibit similar protein profiles and comparable allergen composition, with phospholipase and hyaluronidase activities. The structures of Pol d 1 and Ves v 1 and Pol d 5 and Ves v 5 were highly similar, and the identity levels were high across and within the Polistes and Vespula genera (≥50%). These results suggest the inclusion of venoms from Polistes and Vespula genera as candidates to create a new homologous group for wasp venoms and indicate that the currently described homologous groups require revision. Full article

Background/Objectives: Atopic dermatitis (AD)-related skin inflammation involves the release of cytokines and chemokines from keratinocytes; therefore, keratinocyte-based models are widely used to evaluate the anti-inflammatory potential of botanical extracts. This study examined the relationship between phytochemical profiles and anti-inflammatory potential of baobab fruit 30% and 70% ethanol extracts (BE-30 and BE-70, respectively) in a TNF-α/IFN-γ (TI)-stimulated HaCaT keratinocyte model. Methods: The anti-inflammatory effects of both extracts were evaluated by measuring cytokine and chemokine secretion in TI-stimulated HaCaT cells. Phytochemical characterization was performed using liquid chromatography–tandem mass spectrometry (LC–MS/MS) and targeted high-performance liquid chromatography (HPLC). Results: Both extracts were non-cytotoxic. TI-stimulation markedly increased interleukin (IL)-6, IL-8 and monocyte chemotactic protein (MCP)-1 secretion, while BE-30 and BE-70 significantly reduced all three mediators in a dose-dependent manner. At comparable doses, BE-70 exhibited greater inhibition than BE-30. BE-30 showed a non-monotonic IL-8 response at low concentrations, whereas BE-70 consistently reduced IL-8 in a dose-dependent manner. LC–MS/MS profiling revealed a polyphenol-rich composition, including flavonol glycosides and related phenolic compounds. HPLC confirmed the presence of four marker analytes (procyanidin B2, epicatechin, rutin and tiliroside), which were enriched in BE-70. The content of these four polyphenols was 1.94-fold higher in BE-70. Conclusions: Baobab fruit extracts exhibit anti-inflammatory activity associated with polyphenols. These findings suggest that they could be used as analytical standards and in dermatological applications. Full article