Abstract Thermal processing is essential for liquid milk quality and safety. However, relatively broad standard regulations and inefficient supervision methods encourage enterprises to use excessive heating to maximize commercial profits. Therefore, a pseudo-targeted metabolomics approach based on ultra performance liquid chromatography quadrupole time-of flight mass spectrometry (UPLC-QTOF MS) with multivariate statistical analysis was used to discover biomarkers that can distinguish among pasteurized (29 Pa, 72 °C, 75 °C, and 85 °C), extended shelf life (9 ESL, 121 °C), and ultra-high temperature sterilization (20 UHT, 138 °C) milk. Finally, 10 key metabolites were characterized, including peptides, nicotinamide, N6-methyladenosin, and 2-hydroxycapric acid. The diagnostic performance of these candidate biomarkers was evaluated in 55 validation samples using receiver operating characteristic curve analysis and a non-parametric discrimination model, with 96.4% classification accuracy. These results demonstrated that the pseudo-targeted metabolomics method is reliable and provides effective biomarkers for evaluating the nutritional value of milk subjected to different thermal treatments.

Fenbendazole is a highly effective antiparasitic agent widely used in veterinary medicine; however, its improper application can lead to residues in edible animal-derived products, posing potential health risks to humans through accumulation via the food chain. This study aimed to establish an ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) method for the determination of fenbendazole and metabolites in animal-derived foods. The proposed method involves a one-step solid-phase extraction (SPE) followed by UHPLC-MS/MS analysis for the simultaneous detection of fenbendazole, oxfendazole, and oxfendazole sulfone. The method demonstrated excellent linearity, with correlation coefficients (R²) exceeding 0.99. The limit of detection (LOD) and limit of quantification (LOQ) were 2.0 μg/kg and 5.0 μg/kg, respectively. Average recoveries ranged from 85.20% to 108.42%, with relative standard deviations below 10%. The established method is simple, highly specific, and sensitive, offering a reliable approach for production monitoring, routine batch analysis, and confirmatory testing of fenbendazole and metabolites in animal-derived foods.

Cisplatin (DDP)-induced acute kidney injury (AKI) presents a major challenge in chemotherapy, limiting its clinical utility due to nephrotoxicity. In this study, we explored the therapeutic potential of polyphenol-rich extracts from highland barley (HBPE) in counteracting DDP-induced renal damage. We hypothesized that HBPE could exert protective effects through the modulation of oxidative stress, mitochondrial dysfunction, and metabolic dysregulation. Using UHPLC-QTOF/MS-based untargeted metabolomics, we identified key metabolic disruptions in human embryonic kidney (HEK293) cells treated with DDP, which were substantially reversed by HBPE. The extract significantly attenuated mitochondrial injury and oxidative stress, as shown by decreased malondialdehyde (MDA) levels; increased levels of glutathione (GSH), superoxide dismutase (SOD), and ATP; and reduced reactive oxygen species (ROS) (P < 0.05). Furthermore, HBPE inhibited apoptosis by stabilizing mitochondrial integrity. In vivo, HBPE pretreatment ameliorated renal dysfunction in rats, as evidenced by reduced serum creatinine and blood urea nitrogen (BUN) levels and improved renal histopathology. Metabolomic profiling identified 39 potential biomarkers and revealed that HBPE restored key metabolic pathways, including folate biosynthesis, nicotinate metabolism, and phenylalanine metabolism. These results support the potential of HBPE as a natural nephroprotective intervention during chemotherapy. Derived from a sustainable agricultural source, HBPE offers a promising, low-toxicity strategy for enhancing patient safety and promoting integrative therapeutic development.

Background Honey authentication and traceability are paramount for ensuring product integrity and consumer trust within the global apiculture industry. Authentication focuses on detecting deliberate fraud, such as syrup adulteration, water dilution, and inferior honey blending, which directly undermines product quality and safety. In contrast, traceability aims to verify and confirm the declared botanical, geographical, and entomological origins of honey, which is essential for quality differentiation, protected designation, and supply chain transparency. The limitations of conventional single-dimensional analytical techniques, such as melissopalynology and isotope ratio mass spectrometry, in addressing increasingly sophisticated fraud and complex verification needs necessitate more robust, integrated solutions. Scope and approach This review systematically examines the methodological evolution in both honey authentication and traceability since 2015. For authentication, it details the progression from targeted assays to non-targeted, intelligent screening systems for fraud detection. For traceability, it outlines the advancement from traditional morphological and chemical analyses toward data-driven frameworks for origin verification. The review highlights the paradigm shift toward intelligent systems that integrate food multi-omics with machine learning and deep learning algorithms, which excel at fusing heterogeneous data for accurate analysis. Key findings and conclusions The analysis reveals distinct yet complementary technological pathways for authentication and traceability, driven by their specific objectives. A key trend is the convergence of multi-analytical data sources with advanced computational models to create more powerful and adaptable systems. Future progress hinges on overcoming challenges related to data standardization, model interpretability, and practical deployment. The development of next-generation, integrated frameworks is strategic for reinforcing consumer confidence, ensuring regulatory compliance, and promoting the long-term sustainability of beekeeping.

Thermal processing is essential for milk safety and shelf-life extension but induces complex alterations in milk proteins and peptides that influence product quality and authenticity. In this study, a UPLC-QTOF-MS based peptidomics approach was applied to identify peptide markers reflecting the thermal history of milk across a wide temperature range. Multivariate analysis identified 48 heat-sensitive peptides that effectively discriminated pasteurized, ESL, and UHT milk, while also resolving subtle differences within low-temperature pasteurization (72-85°C). These peptides mainly originated from the N- or C-terminal regions of αS1-, β-, and κ-caseins. Marker peptide formation was independent of milk fat content. In contrast, reactive oxygen species oxidation experiments demonstrated a synergistic contribution of oxidative pathways to the formation and modification of a subset of peptides, particularly those in oxidation, pyro-prone hydrophobic regions. Overall, this study establishes a robust peptidomics framework for thermal process authentication and provides mechanistic insight into heat-induced peptide generation in milk.

Introduction In the environment, mycotoxins and fungicides frequently coexist, potentially causing synergistic risks to organisms. Epoxiconazole (EPO) and aflatoxin B1 (AFB1) are a common fungicide and mycotoxins, respectively, which are widely present in the environment and have toxic effects on multiple organs once entering the organism, but it is still unclear whether the co-exposure has a synergistic toxic effect. Objectives This study delves into the molecular mechanisms underlying the co-exposure to EPO and AFB1, emphasizing multi-organ toxicity in female zebrafish (F0 generation) and potential transgenerational impacts on the offspring embryos (F1 generation) through multi-omics approaches. Results Findings indicate that exposure to either EPO or AFB1, individually or combined, intensified intestinal pathological damage, decreased the expression of tight junction proteins, altered gut microbiota composition, and induced intestinal inflammation, with co-exposure causing more severe effects. RNA-seq analysis revealed an enrichment of ferroptosis and apoptosis pathways in the liver and ovaries of F0 zebrafish. Co-exposure markedly altered the expression of associated molecules, exacerbating pathological damage in these organs. Molecular docking studies revealed that AFB1 exhibited lower binding energies to Caspase3, GPX4 and IL-1β compared to EPO, suggesting that it may have a higher binding capacity. Furthermore, both single and combined exposures modified the expression of molecules related to apoptosis, inflammatory response, and ferroptosis in unexposed F1 embryos, with co-exposure demonstrating more significant biological effects, thereby confirming transgenerational toxicity. Conclusion The present study provides preliminary evidence on the potential mechanisms of combined exposure-induced multi-organ toxicity, highlighting ferroptosis of the liver and apoptosis of the ovary as key pathways. These findings provide new perspectives and methods for risk assessment of multiple environmental pollutants.

Food allergies constitute a significant and escalating global public health issue. Over the past decade, efforts have intensified to prevent and treat these allergies, including exploring new anti-allergic agents and natural bioactive compounds with minimal side effects. Despite progress, consensus on effective strategies remains elusive. This study undertakes a comprehensive review and discussion of current anti-allergic medications, natural bioactive ingredients, and innovative nano/micro-carriers, focusing on four key mechanisms: binding to allergen epitopes, modulating gut microbiota, restoring intestinal epithelial integrity, and regulating immune responses. Many natural compounds show effectiveness through multiple pathways. Advances in nanotechnology have improved delivery systems such as nanoparticles and sporopollenin exine capsules, enhancing targeted delivery and efficacy. Given the distinct cellular and molecular targets of these anti-allergic agents, investigating synergies between natural and synthetic drugs is essential. For instance, combine traditional anti-allergic drugs with glucocorticoids to quickly relieve initial symptoms; then, use natural agents like probiotics for immune regulation, reducing treatment time and recurrence risk. This review lays the foundation for a scientific framework to guide the future development of combination therapy models in clinical applications.

Edible insects, exemplified by honeybee pupa, represent a sustainable protein source but are associated with undercharacterized allergenic risks. This study innovatively addresses this gap by targeting honeybee pupa-derived Profilin, a cross-reactive pan-allergen, through conjugation with caffeic acid phenethyl ester (CAPE), a bioactive polyphenol derived from propolis. Structural and biochemical analyses revealed that CAPE-induced covalent polymerization and conformational remodeling (characterized by a shift from α-helix dominance to β-sheet dominance, accompanied by tertiary structural perturbation) resulted in about 60 % suppression of IgE-binding capacity compared to native Profilin. Notably, CAPE-conjugated honeybee pupa protein exhibited dual functionality: antioxidant activity increased 18-fold for ABTS+ scavenging and 2-fold for DPPH· inhibition, while murine allergy models showed about 70 % reduction in allergic indices and attenuated splenic inflammation. By mechanistically linking protein-polyphenol interactions to hypoallergenicity and nutraceutical enhancement, this work establishes an engineering framework for safer insect-based foods, bridging critical gaps in alternative protein safety and functional food innovation.

In this study, a new type of carbon dots based on diethyl ferulate (DEF-CDs) was developed, and doped into polyvinyl alcohol (PVA) to prepare an active food packaging film that effectively inhibits photooxidation and microbial contamination of food. Benefit from the rational design of the structure and optimization of the precursor for the DEF-CDs synthesis, 0.4 % of DEF-CDs in PVA film can prevent 100 % ultraviolet light (UV) and 99.90 % high-energy blue light (HEBL). Furthermore, its UV and HEBL blocking capacity shows no significant decline under 40 days of accelerated aging tests. Moreover, the DEF-CDs/PVA film exhibits significant antibacterial capacity, and with the concentration of DEF-CDs increased from 0 % to 0.8 %, the inhibition zone diameters increased from 8 mm to 22.8 mm for Staphylococcus aureus and from 8 mm to 11.8 mm for Escherichia coli O157:H7. Finally, using the DEF-CDs/PVA film as active food packaging, the shelf life of strawberry, jujube and milk was extended significantly. This work exhibits the potential of DEF-CDs as multifunctional nano- reinforcements for constructing protective food packaging.

Background The complexity of contemporary food systems requires sophisticated risk-benefit assessment (RBA) methodologies to ensure food safety, optimize nutritional adequacy, and sustainability, while addressing personalized needs. Traditional RBA frameworks face challenges with innovations such as personalized nutrition and novel food technologies, necessitating a more adaptive approach that considers individual health outcomes with broader environmental and societal goals. Scope and approach This review critically examines recent advancements and methodological trends in RBA, emphasizing the integration of multi-tiered RBA approaches and personalized nutrition strategies. It examines the incorporation of personalized nutrition into RBA framework, which customizes dietary recommendations based on genetic, microbiome, and lifestyle factors. The review explores risk-benefit communication as a critical bridge connecting scientific assessment with practical implementation, particularly at the intersection of personalized nutrition and sustainable diets. Additionally, it explores the integration of environmental and economic sustainability considerations into RBA through nutritional life cycle assessment (nLCA), which provides a methodological foundation for evaluating environmental impacts alongside nutritional adequacy. Through diverse case studies and practical applications, this review establishes a holistic RBA framework that aligns individual health optimization with societal goals. Key findings and conclusions The review identifies critical challenges, including data gaps, methodological limitations in evaluating complex dietary interactions, and the necessity for improved health metrics that capture the multifaceted effects of dietary patterns. It proposes strategic frameworks for sustainable diet integration in RBA, emphasizing dietary pattern transitional analysis. Additionally, it advances multidimensional indicator development through integration of cross-dimensional metrics. Future directions suggest enhancing data infrastructure, fostering interdisciplinary collaboration, and refining integrative health metrics to advance RBA's role in advocating for health-promoting and environmentally sustainable diets.

Milk pasteurization and sterilization by heat treatment have an exciting history, which followed steady steps. The main aim of these treatments is to extend the shelf life of milk by destroying pathogenic and milk spoilage bacteria. With developments in pasteurization techniques, the assurance of milk safety, and extended shelf life, pasteurized bovine milk has become a staple food, especially in Western diets. However, some concerns have recently been raised about the effect of pasteurization on the sensory properties and nutritional quality of milk, and alternative methods, such as high-pressure processing, are being investigated. The primary purpose of milk pasteurization and sterilization is summarized in this review article. The associated changes that affect the compositional, sensory, and nutritional quality of milk are discussed, with particular emphasis on protein structure and function. The review is concluded by considering alternative methods, their advantages and limitations, along with future prospects.

Multi-targeted tyrosine kinase inhibitor QLNC-3A6 Di-maleate, a structurally novel small molecule compound, has therapeutic efficacy for the treatment of canine cutaneous mast cell tumor (CMCT) caused by mutations in the c-Kit gene. Since pharmacokinetic (PK) information plays an important role in the development and application of new drugs, etc., a rapid, highly sensitive and selective UHPLC-MS/MS analytical method was developed and validated for the first time in this study for the quantitative detection of QLNC-3A6 in canine plasma. 100 mu L of plasma was precipitated using 350 mu L of acetonitrile, and Chromatographic separation was performed on a Phenomenex Kinetex C18 column (50 x 2.1 mm, 2.6 mu m) at a flow rate of 0.4 mL/min, the mobile phases were set to 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile (B). The calibration curve linear range was 0.5-100 ng/mL (R-2>0.99). The intraday and interday precision values (relative standard deviation, RSD) were 2.06-13.57% and 6.90-9.14%. Intraday and interday accuracies were -10.73 to 9.54% and -3.86 to 0.70% respectively. The dilution integrity RSD value and stability RSD value were less than 3.77 and 7.45%, respectively. Subsequently, the pharmacokinetics were investigated in canine after oral administration of QLNC-3A6 Di-maleate tablets at a dose of 3 mg/kg BW using this method. The results showed that QLNC-3A6 showed fast absorption rate, rapid distribution and slow metabolic elimination in canine plasma. The results of the main PK parameters including lambda z, T1/2 lambda z, C-max, T(max )and AUC(last) were 0.07 +/- 0.01/h, 11.00 +/- 2.57 h, 50.88 +/- 31.94 ng/mL, 9.08 +/- 11.57 h and 836.48 +/- 230.53 ng h/mL, respectively.

Introduction Photo-oxidation is recognized as a contributor to the deterioration of milk quality, posing potential safety hazards to human health. However, there has been limited investigation into the impact of consuming photo-oxidized milk on health. Objectives This study employs multi-omics analysis techniques to elucidate the mechanisms by which photo-oxidized milk induces oxidative stress in the liver. Methods Mouse model was used to determine the effect of the gavage administration of milk with varying degrees of photo-oxidation on the mouse liver. The damage degree was established by measuring serum markers indicative of oxidative stress, and with a subsequent histopathological examination of liver tissues. In addition, comprehensive metabolome, lipidome, and transcriptome analyses were conducted to elucidate the underlying molecular mechanisms of hepatic damage caused by photo-oxidized milk. Results A significant elevation in the oxidative stress levels and the presence of hepatocellular swelling and inflammation subsequent to the gavage administration of photo-oxidized milk to mice. Significant alterations in the levels of metabolites such as lumichrome, all-trans-retinal, L-valine, phosphatidylglycerol, and phosphatidylcholine within the hepatic tissue of mice. Moreover, photo-oxidized milk exerted a pronounced detrimental impact on the glycerophospholipid metabolism of mice liver. The peroxisome proliferator-activated receptors (PPAR) signaling pathway enrichment appreciated in the animals that consumed photo-oxidized milk further supports the substantial negative influence of photo-oxidized milk on hepatic lipid metabolism. Gene set enrichment and interaction analyses revealed that photo-oxidized milk inhibited the cytochrome P450 pathway in mice, while also affecting other pathways associated with cellular stress response and lipid biosynthesis. Conclusion This comprehensive study provides significant evidence regarding the potential health risks associated with photo-oxidized milk, particularly in terms of hepatic oxidative damage. It establishes a scientific foundation for assessing the safety of such milk and ensuring the quality of dairy products.

Food allergens are mainly naturally-occurring proteins with immunoglobulin E (IgE)-binding epitopes. Understanding the structural and immunogenic characteristics of allergenic proteins is essential in assessing whether and how food processing techniques reduce allergenicity. We here discuss the impacts of food processing technologies on the modification of physicochemical, structural, and immunogenic properties of allergenic proteins. Detection techniques for characterizing changes in these properties of food allergens are summarized. Food processing helps to reduce allergenicity by aggregating or denaturing proteins, which masks, modifies, or destroys antigenic epitopes, whereas, it cannot eliminate allergenicity completely, and sometimes even improves allergenicity by exposing new epitopes. Moreover, most food processing techniques have been tested on purified food allergens rather than food products due to potential interference of other food components. We provide guidance for further development of processing operations that can decrease the allergenicity of allergenic food proteins without negatively impacting the nutritional profile.

With development of modern food industry, plant-based milk products are widely used to replace dairy products to cook different kinds of food. Due to different origins, it is necessary to assess the nutrition difference between dairy and plant-based milk products. Phospholipids and unsaturated fatty acids, as the important nutrients in cream and butter, were closely related to body development and health. This study developed a rapid determination of phospholipids and double bonds in creams and butters based on 31P NMR and 1H NMR for nutritional difference assessment. Unsupervised principal component analysis and supervised orthogonal least squares discriminant analysis showed marked differences between dairy and plant-based milk products. Eight compounds were screened by variable importance in projection, fold change and P-value. Furthermore, two biomarkers (phosphatidylserine and phosphatidylethanolamine) were identified to distinguish dairy and plant-based milk products. The study demonstrated that 31P NMR has great potential for rapidly distinguishing milk origins.

Abuse of glucocorticoid veterinary drugs in dairy industry can potentially threat milk safety and consequently influence human health. Here a reliable method for determination of 58 glucocorticoid drug residues in milk was established by combining solid phase extraction with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The analytes were extracted with acetonitrile and cleanup with EMR-Lipid lipid removal column. The analytes were chromatographically separated using Poroshell EC-C18 column and acquired by electrospray ionization with multiple-reaction monitoring (MRM) mode. The limit of quantification (S/N ≥ 10) ranged from 0.2 to 2.0 µg/kg and the limit of detection (S/N ≥ 3) ranged from 0.1 to 1.0 µg/kg. Average recoveries were from 71% to 113%, the relative standard deviations (RSDs) were less than 15%, and the correlation coefficients (R2) of calibration curves exceeded 0.99. The method was applied to detect twenty milk products obtained from local supermarkets including ten pasteurized milk and ten UHT milk. Two endogenous glucocorticoids, i.e. hydrocortisone and cortisone were detected but not exceed the maximum residue limits (MRLs).

Milk proteins are important components that confer several nutritional and functional properties to milk and dairy products. As these proteins have complex nature and wide variability, their analysis is not trivial and requires several methods to be used individually or in combination. In this article, we provide a comprehensive review of conventional and more advanced methods used in the analysis of milk protein content, composition, and structural properties. Different traditional methods (mainly titration and spectrophotometric) are still used for the determination of total protein content, while chromatographic and electrophoretic techniques, enzyme-linked immunosorbent assays, and infra-red spectroscopy are used to separate, identify, and quantify individual proteins. For advanced structural identification of separated proteins, nuclear magnetic resonance, X-ray diffraction, and mass spectroscopic techniques can be used. Of these methods, the analyst may select those relevant to different applications in milk and dairy products research.

Photooxidation is one of the main causes of the deterioration of milk quality during processing and marketing. This study aimed to investigate the variation in peptides after photooxidation using peptidomic techniques, and how cow species, oxygen content, and light intensity affect photooxidation. The different peptides were identified and quantified using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Eighteen milk samples were subjected to light treatment. Seven types of peptides were identified as photooxidation markers. Subsequently, the effects of milk variety, oxygen content, and light intensity on photooxidation were studied, and sensory evaluations were performed. Dairy cow breed, oxygen content, and light intensity all affect photooxidation. Sensory evaluation verified that light and oxygen are necessary for the photooxidation of milk. The peptide m/z+ 529.2783 (LLDEIKEVV), both in different varieties of milk and in different brands of commercially available milk, showed a large variation in multiplicity, and its content was closely related to oxygen and light. This peptide was not produced in the absence of oxygen and light, and its relative content increased with the duration of light exposure. These results suggest that the peptidomics method is an effective tool for distinguishing between normal and photooxidized milk.

This study investigated the changes in milk powder on spray and freeze drying by using untargeted metabolomics. The metabolome change of the spray-dried milk powder was also studied during storage at 20 °C, 30 °C and accelerated storage at 40°C. This study demonstrated that spray drying process is an efficient way to produce milk powder and extend its shelf life. However, some metabolite changes (e.g. uridine, riboflavin, adenine, peptides etc.) were observed during this process. The milk powder stored at 20 °C and 30 °C exhibited a stable profile for up to 336 days. However, the milk powder stored at 40 °C showed significant changes from day 84 and onwards, with increasing levels on organic acids (e.g. orotic acid, 2-ketobutyric acid, acetic acid, citraconic acid and oxoglutaric acid), amino acids (e.g. creatinine, carnitine, N-acetyl-L-phenylalanine, glutamic acid, proline, tyrosine, alanine, creatine, tryptophan), peptides (2~4 amino acids) and carbohydrate derivatives (e.g. N-acetyllactosamine, glycerol 3-phosphate, D-ribose 5-phosphate and raffinose) and reducing contents on some fatty acids, such as PE (14:0/0:0), lauric acid, MG, caprylic acid, capric acid et al. This research not only provides a full understanding of the milk metabolomic changes in spray drying and storage but also provides a list of potential markers that could be used to track the shelf life of milk powders.