Ensuring food safety requires robust screening approaches capable of detecting the administration of growth-promoting agents in livestock. While previous metabolomics studies have demonstrated proof-of-concept, most were limited to a single class of compounds and offered only partial metabolome coverage due to the use of individual analytical platforms. The present study aimed to develop a comprehensive classification model through a more global exploration of the metabolome. This integrative workflow represents a major step towards implementing effect-based metabolomics screening within official food control frameworks. A total of 502 urine samples from six experiments (n = 59 cattle) involving β-agonists, steroids, and SARMs were analysed using four complementary LC-HRMS platforms. This multi-platform approach expanded metabolome coverage by capturing a wide range of molecular fingerprints. Multiblock Consensus-OPLS analysis was applied to integrate the four datasets into a joint modelling strategy, thereby enhancing data interpretation. This strategy provided an innovative framework that increased the predictive power of the classification model and underscored the complementarity of the LC-HRMS techniques. Overall, this comprehensive workflow enabled the efficient classification of samples from animals subjected to multiple anabolic treatments within a single analysis. Such advances not only strengthen detection capabilities but also offer a versatile tool to address key public health concerns.

Chemical contaminants are widely dispersed in the environment in all its dimensions, posing significant public health problems. Comprehensive knowledge of these stressors is a prerequisite for assessing the associated risk and implementing public policy measures to reduce the level of population exposure. Nontargeted suspect screening approaches broaden the knowledge of the chemical human exposome. We developed and used a suspect screening method based on large spectral libraries. Chemical profiling was based on a combined LC- and GC-HRMS approach. The methodology was applied to 16 samples spanning the environment, food, and health continuum. Using a combination of matching and scoring data, a total of 547 compounds were likely identified, from which the chemical structure of 63 molecules was confirmed to the highest level of certainty. Wastewater, and more generally environmental samples, had the highest number of chemicals detected, while fish samples had a lower number. Pharmaceuticals, pesticides, and personal care product-related compounds were found to be the most common compounds in and between the extracts, particularly in water and serum samples. Many natural and endogenous compounds were consistently annotated in the samples submitted for analysis, regardless of the compartment investigated.

The recombinant bovine somatotropin (rbST) is a synthetic hormone developed to mimic the effects of the endogenous growth hormone, also known as bovine somatotropin (bST). Although rbST use in dairy cows is authorized in several countries, it is currently banned in Europe. Different methods for screening and confirmatory detection of rbST were developed, mainly based on LC-MS/MS and immune-enzymatic assays. However, some commercial forms of rbST have above the same amino acid sequence of bST, making it difficult to produce a reliable differentiation of recombinant from endogenous forms. Complementary strategies for indirect detection of rbST can therefore be considered as alternative biomarker-based tools. Untargeted transcriptomics was applied to characterize the microRNAs (miRNA) isolated from milk extracellular vesicles (EVs) in rbST-treated animals, aiming the identification of non-coding biomarkers related to its administration. Sequencing analysis of 63 archive samples collected during previous animal trial allowed for the identification of 35 differentially expressed (DE) miRNAs. A validation study performed by qPCR on a further 70 milk samples from a field survey confirmed the significant upregulation of bta-miR-10167-3p in milk EV from rbST-treated cows. The results obtained suggest the potential use of bta-miR-10167-3p as a non-invasive biomarker to be considered in novel screening strategies, needed to tackle rbST misuse in dairy cows.

Dietary intake has been highlighted as the main route of human exposure to polychlorinated n-alkanes (PCAs). In this study, we investigated the dietary exposure of the French population to PCAs by analysing 34 vegetable oil and margarine samples as well as in 50 foods of animal origin, often reported as the most contaminated food items in the human diet. Levels were determined using both LC/ESI-HRMS and GC/ECNI-HRMS couplings, for further comparison. For vegetable oil samples, precisions, trueness and uncertainties were < 15 %, ≤22 % and 7-25 %, respectively. For foods of animal origin, higher variability was observed between short- and medium-chain PCAs, as well as between instruments. Average estimated daily intakes were 7.9 and 100 ng per kg of body weight per day for ∑PCAs-C10-13 and ∑PCAs-C14-17, respectively. Preliminary risk characterisation indicated no health concern for the considered food categories as regards ∑PCAs-C10-13 and ∑PCAs-C14-17.

Ensuring food safety is a shared priority for both the European Union (EU) and China, yet differences in regulatory frameworks, enforcement mechanisms, and risk assessment approaches present challenges for harmonization. The SAFFI project (2020-2024), which concluded recently, seeked to bridge these gaps by fostering collaboration in food safety management, with a specific focus on infant food safety. Bringing together partners from several EU member states and Chinese provinces, SAFFI aimed to improve hazard identification, risk ranking, detection methods, and mitigation strategies for both microbial and chemical contaminants in infant food supply chains. This article first provides a comparative analysis of EU and Chinese food safety regulations, highlighting key similarities and differences in legal structures, risk assessment procedures, and enforcement strategies. Infant food safety standards are particularly strict in both regions, yet inconsistencies in testing methods, contaminant limits, and monitoring approaches necessitate greater alignment. The second part of the article details the scientific advances achieved through SAFFI to support regulatory convergence. These include structured databases and decision-support tools for hazard identification and prioritization, improved processing and preservation techniques for contaminant control, and cutting-edge analytical methods for detecting and monitoring known and emerging chemical and microbial hazards. The project also examined the role of consumer practices in mitigating risks and explored pathways for integrating rapid detection technologies into food safety systems. Overall, the outcomes of SAFFI contribute to enhanced consumer protection, the promotion of safe international trade, and the development of more coherent, science-based food safety policies through sustained EU-China cooperation.

Polyvinyl chloride (PVC) is one of the most widely used polymers, which contrasts with the low amount of literature on the leaching of associated additives and in particular unintended oligomers, a class of non-intentionally added substances (NIAS). This study sheds light on the occurrence of vinyl chloride oligomers (VCOs) in a variety of PVC analytical standards (n = 4), PVC items employed for construction, medical or food contact applications (n = 14), as well as in foodstuffs and environmental matrices (n = 10). Samples were analysed by liquid chromatography coupled to high-resolution mass spectrometry hyphenated with chloride-enhanced electrospray ionisation. Series of saturated (0VCO), monounsaturated (1VCO), diunsaturated (2VCO) and triunsaturated (3VCO) VCOs were revealed. The dominant series remained 1VCOs of the general formula C2nH3nCln, accounting for ∼80 % of VCOs in PVC standards. A risk of signal overlap was found between 0VCOs of the general formula C2nH3n+1Cln+1 (accounting for ∼8 % in PVC standards) and polychlorinated alkanes making up chlorinated paraffins, given that they share the same chemical formulas. A methodology for discriminating between signals arising from VCOs and chlorinated paraffins has been proposed. VCOs were detected in 88 % of the samples analysed (other than standards), and in particular in some foodstuffs and environmental matrices, suggesting that VCOs have the capacity to leach out of PVC materials, and thus contaminate food and the environment. Overall, these results call for greater attention to be paid to vinyl chloride oligomers and raise the question of whether they pose a risk to living organisms.

Per- and polyfluoroalkyl substances (PFASs) are increasingly recognized as emerging contaminants, with some classified as persistent organic pollutants (POPs) and four recently regulated in food in the EU. However, due to the large structural diversity of PFASs, comprehensive monitoring remains essential. This study aimed to develop and apply a suspect and non-targeted screening approach to identify PFASs beyond those conventionally monitored in foodstuffs. The methodology combined optimized QuEChERS sample preparation, LC-HRMS acquisition, and prioritization of fluorinated signals, applying both suspect screening (SS) and non-targeted screening (NTS) strategies. Compared to targeted solid-phase extraction (SPE) coupled with MS/MS (QqQ) acquisition, this approach detected a broader range of PFASs in various food samples. Notably, as one of the first studies to apply NTS-a method typically used in environmental analysis-to food, it demonstrated the ability to detect both known and previously unlisted PFASs, such as PFPrA in an egg sample. This expanded approach enhances exposure assessment and supports the implementation of HRMS-based strategies for regulatory control and risk assessment.

Per- and polyfluoroalkyl substances (PFAS) are persistent, potentially harmful synthetic chemicals. While they can accumulate in foodstuffs, current monitoring often targets only a few compounds, likely underestimating dietary exposure. In this study, 58 food samples from Europe and North Africa-including commercial products and items from known European contamination hotspots-were analyzed using a validated high-resolution mass spectrometry workflow combining suspect screening (SS) and non-targeted screening (NTS). Seventeen PFAS were confirmed through SS, with up to 15 different PFAS in fish samples from hotspots. While NTS revealed four additional fluorinated substances: Perfluoropropanoic acid (PFPrA) detected in 48 % of samples, 6:2 Fluorotelomer sulfonic acid (6:2 FTS), Fipronil, and Fipronil sulfone. These results highlight the geographical variability of PFAS contamination in food and demonstrate the value of combined SS/NTS approaches in identifying both known and emerging PFAS, supporting more comprehensive, regulation-aligned risk assessments.

This review explores the challenges Arctic populations face regarding contaminant exposure and its intricate relationship with traditional diets. Particular attention is given to the unique situation of northern women, whose dietary habits influence their exposure to these contaminants. Despite the recent and rapid dietary shift towards a more “westernized” diet, traditional foods remain a cornerstone of Arctic communities’ sustenance. However, the consumption of such foods, particularly marine mammals, has been consistently associated to elevated levels of lipophilic contaminants including persistent organic pollutants (POPs) and trace elements. Notable gendered differences emerge in dietary patterns, with northern women reportedly consuming fewer traditional products, thereby reducing their contaminant exposure. Additionally, women of childbearing age benefit from unique elimination pathways––through pregnancy, breastfeeding, and menstruation––that men lack. This combination of lower traditional food intake and the existence of gender-specific elimination routes has resulted in a lower contaminant burden in women compared to men. For instance, blood concentrations of organic contaminants such as polychlorinated biphenyls, organochlorine pesticides, per- and polyfluoroalkyl substances, and trace elements like lead, are on average 20 to 40% lower in women. Nevertheless, these lower contaminant levels do not necessarily imply reduced health risks, as women’s susceptibility to these substances may differ markedly from that of men.

This paper examines the potential impact of introducing new hazard detection, control, and mitigation tools in the infant food chain by conducting a qualitative and stakeholder-driven benefit-risk analysis of prospective scenarios. The scenarios envision the possible implementations of the tools and approaches developed in the 'Safe Food For Infants' (SAFFI) Europe-China project, which include detection technologies, hazard control and mitigation strategies, as well as models for hazard identification and risk ranking. The objective is to evaluate how the implementation of these tools could affect food safety management, particularly in light of diverse consumer behaviors, evolving technologies, and regulatory contexts. Through a scenario-building approach, the paper constructs potential futures for the infant food chain, followed by a detailed benefit-risk analysis. The analysis incorporates a variety of stakeholder perspectives and criteria, including safety, economic impact, and technological advancements. The findings provide valuable insights into how different strategies for hazard control and mitigation may improve food safety, while also considering the trade-offs associated with these strategies. The paper ultimately proposes a global ranking of scenarios based on a collective attitude score, offering a comprehensive evaluation of prospective futures for infant food safety.

Non-targeted and suspect screening analysis using liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) holds great promise to comprehensively characterize complex chemical mixtures. Data preprocessing is a crucial part of the process, however, some limitations are observed: (i) peak-picking and feature extraction might be incomplete, especially for low abundant compounds, and (ii) limited reproducibility has been observed between laboratories and software for detected features and their relative quantification. We first conducted a critical review of existing solutions that could improve the reproducibility of preprocessing for LC-HRMS. Solutions include providing repositories and reporting guidelines, open and modular processing workflows, public benchmark datasets, tools to optimize the data preprocessing and to filter out false positive detections. We then propose harmonized quality assurance/quality control guidelines that would allow to assess the sensitivity of feature detection, reproducibility, integration accuracy, precision, accuracy, and consistency of data preprocessing for human biomonitoring, food and environmental communities.

The chemical burden on the environment and human population is increasing. Consequently, regulatory risk assessment must keep pace to manage, reduce, and prevent adverse impacts on human and environmental health associated with hazardous chemicals. Surveillance of chemicals of known, emerging, or potential future concern, entering the environment-food-human continuum is needed to document the reality of risks posed by chemicals on ecosystem and human health from a one health perspective, feed into early warning systems and support public policies for exposure mitigation provisions and safe and sustainable by design strategies. The use of less-conventional sampling strategies and integration of full-scan, high-resolution mass spectrometry and effect-directed analysis in environmental and human monitoring programmes have the potential to enhance the screening and identification of a wider range of chemicals of known, emerging or potential future concern. Here, we outline the key needs and recommendations identified within the European Partnership for Assessment of Risks from Chemicals (PARC) project for leveraging these innovative methodologies to support the development of next-generation chemical risk assessment.

Bromochloro alkanes (BCAs) have been manufactured for use as flame retardants for decades, and preliminary environmental risk screening suggests they are likely to behave similarly to polychlorinated alkanes (PCAs), subclasses of which are restricted as Stockholm Convention Persistent Organic Pollutants (POPs). BCAs have rarely been studied in the environment, although some evidence suggests they may migrate from treated-consumer materials into indoor dust, resulting in human exposure via inadvertent ingestion. In this study, BCA-C14 mixture standards were synthesized and used to validate an analytical method. This method relies on chloride-enhanced liquid chromatography-electrospray ionization-Orbitrap-high resolution mass spectrometry (LC-ESI-Orbitrap-HRMS) and a novel CP-Seeker integration software package for homologue detection and integration. Dust sample preparation via ultrasonic extraction, acidified silica cleanup, and fractionation on neutral silica cartridges was found to be suitable for BCAs, with absolute recovery of individual homologues averaging 66 to 78% and coefficients of variation ≤10% in replicated spiking experiments (n = 3). In addition, a total of 59 indoor dust samples from six countries, including Australia (n = 10), Belgium (n = 10), Colombia (n = 10), Japan (n = 10), Thailand (n = 10), and the United States of America (n = 9), were analyzed for BCAs. BCAs were detected in seven samples from the U.S.A., with carbon chain lengths of C8, C10, C12, C14, C16, C18, C24 to C28, C30 and C31 observed overall, though not detected in samples from any other countries. Bromine numbers of detected homologues in the indoor dust samples ranged Br1-4 as well as Br7, while chlorine numbers ranged Cl2-11. BCA-C18 was the most frequently detected, observed in each of the U.S.A. samples, while the most prevalent degrees of halogenation were homologues of Br2 and Cl4-5. Broad estimations of BCA concentrations in the dust samples indicated that levels may approach those of other flame retardants in at least some instances. These findings suggest that development of quantification strategies and further investigation of environmental occurrence and health implications are needed.

Bromochloro alkanes (BCAs) are a class of flame retardants similar in structure to polychlorinated alkanes (PCAs), which are the major component of short-chain chlorinated paraffins (SCCPs) listed as Persistent Organic Pollutants under the Stockholm Convention. BCAs have recently been detected for the first time in environmental samples. Due to the complete lack of commercially available analytical standards, no method for quantifying BCAs has been reported to date. In this study, 16 custom-synthesised standards with mixed bromine and chlorine halogenation and carbon chain lengths ranging from C10 to C17 were characterized by liquid chromatography and Orbitrap high-resolution mass spectrometry and used to assess the applicability of pattern deconvolution quantification strategies for BCAs in indoor dust. Br1-9 and Cl1-8 BCAs were detected as [M + Cl]- adduct ions among the C10 to C17 standards, as well as numerous PCA homologues. After applying correction factors to account for the presence of PCAs in the standards, triplicate fortification experiments using varied halogenation composition and concentration determined an average measurement accuracy of 81% over the carbon chain lengths studied and coefficient of variance ≤20% between replicates. Overall, approximately 89% of the ΣBCA concentrations quantified in the fortification trials met the European Union Reference Laboratory's accuracy acceptability criteria recommended for PCAs, between 50 and 150%. Application of the BCA pattern deconvolution quantification procedure to seven representative indoor dust samples from the United States of America revealed a low correlation between the homologue distribution in the samples and the prototype standards (R2 ≤ 0.40), which precluded reliable quantification. This study indicates that pattern deconvolution is an appropriate strategy for quantifying BCAs in environmental samples, but that a large set of appropriate mixture standards will be required before more reliable estimates of BCA concentrations can be achieved in indoor dust.

A Physiology Based Pharmacokinetic (PBPK) model has been developed to predict the kinetics of Persistent Organic Pollutants (POPs) in laying hens. Different datasets have enabled the calibration of the model for chlordecone (CLD), an organochlorine pesticide used in the French West Indies between 1972 and 1993, as well as for chlorinated paraffins (CPs), widely used for various industrial applications worldwide. For this purpose, the sensitivity analysis showed that intake parameters, laying rate, partition coefficients of yolk, hepatic clearance, percentage of metabolism and age were key parameters. Applied to CLD and CPs, this model shows a good capacity for prediction, with 88 % of the experimental values ranging within 1.5-fold of the predicted value at steady state for CPs and 100 % for CLD. The fine modelling of the physiology and the laying process contributes to precision of the model and gives genericity, enabling the switch from one bird species to another. The model can be implemented with other POPs if the clearance and partition coefficient are known.

Exposure to polychlorinated biphenyls (PCBs) has been linked to dyslipidemia. Under acute exposure to PCBs, it has been observed that the secretion of bile acids (BAs) can be impacted, limiting (indirectly) lipid absorption in the gut. In this context, two non-targeted metabolomics studies on pig serum have recently suggested that BA concentrations may fluctuate under exposure to current non-dioxin-like (NDL)-PCB levels in food, reflecting the acute effects of such chronic exposure. The objective of this research is to implement a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for BA analysis in order to validate the findings of previous metabolomics studies, in which BA levels in serum samples from pigs exposed to environmental doses of NDL-PCBs were highlighted to be affected. The proposed LC-MS method involves the use of a C18-pentafluorophenyl LC column, which is not usually selected for the separation of BAs, but shows better performance for the separation of isomers than typical C18 columns. This LC-MS method shows excellent analytical performance such as low limits of detection (LODs) (<= 1 ng/mL for most BAs) and good linearity (R-2 > 0.994), while no matrix effect was observed. A total of 13 BAs have been quantified, while further BA isomers could be detected and semi-quantified. The application of this targeted LC-MS method confirmed previous findings, suggesting that exposure to low doses of NDL-PCBs decreases the concentration of BAs (i.e., glycochenodeoxycholic acid, hyodeoxycholic acid and taurochenodeoxycholic acid) while the effect on the precursors (cholic acid and chenodeoxycholic acid) is less pronounced.

Despite the growing interest in PCNs and the dioxin-like toxicity exhibited by a number of congeners, a comprehensive assessment of their contribution to the cocktail of dioxin-like contaminants is still lacking. To address such a shortcoming, this study investigated the PCN contamination in foodstuffs recently acquired in France, together with that of the regulatory polychlorinated dibenzodioxins/furans (PCDD/Fs) and polychlorinated biphenyls (PCBs). PCNs were ubiquitous at levels (& sum;70 PCNs = 2.5-150 pg g(-1) wet weight) similar to those reported in other countries, with maximum concentrations observed in fish and fishery products from the North-East Atlantic Ocean. Their congener patterns further suggested unintentional releases of PCNs, while those of the other foodstuffs were correlated to the historical PCN profiles. Low risk from dietary exposure was estimated (& sum;70 PCNs-EDIs of 60-360 pg kg(-1) bw d(-1), & sum;24 PCNs-TEQ-EDIs of 8 x 10(-3)-2.2 x 10(-2) pg TEQ kg(-1) bw d(-1)), with milk and dairy products being the highest contributors, followed by meat and meat products. Finally, the rather high contributions of PCNs to the total PCNs+PCDD/Fs+PCBs concentrations (0.9-50%, average of 9%) and the toxic equivalents (0.2-24%, average of 5%) show that these substances are not minor components of the PCNs+PCDD/Fs+PCBs cocktail.

Background: Hydrophilic interaction liquid chromatography (HILIC) and capillary electrophoresis (CE) coupled to mass spectrometry (MS) are two of the most widely used techniques to explore and characterize the polar metabolome. Both platforms have some peculiar characteristics that make them more suitable for studying different kind of metabolites and different parts of the metabolome. Consequently, their combined use is recommended to extend the coverage of the polar metabolome, particularly when multiple metabolic pathways are expected to be perturbed, as in toxicological studies related to exposure to environmental pollutants such as polychlorinated biphenyls (PCBs). It is unclear whether chronic exposure to low doses leads to effects on the most polar metabolites. Results: In this work, both HILIC-MS and CE-MS platforms have been used to characterize the polar metabolome of blood serum from pigs exposed to low-doses (20 ng/kg body weight (b.w.)/day) of Aroclor 1260 (a technical mixture of PCBs). The use of both platforms aims to cover large parts of the polar metabolome to better characterize the effects of PCB exposure. The results showed that the combined use of HILIC-MS and CE-MS effectively extended the coverage of the metabolome and the interpretability of the data, highlighting their complementarity in metabolic pathways analysis. The analysis of the polar metabolome evidenced a strong effect of exposure on amino acid metabolism, affecting 10 of the 20 proteic amino acids, and a consistent overflow in the urea cycle. Significance: This work highlights the need to implement analytical multi-platforms to better address metabolomics studies, showing the complementarity of two widely used analytical techniques (i.e. HILIC-MS and CE MS) to study the polar metabolome in a risk assessment framework.

Steroids are cholesterol-derived biomolecules that play an essential role in biological processes. These substances used as growth promoters in animals are strictly regulated worldwide. Targeted assays are the conventional methods of monitoring steroid abuse, with limitations: only detect known metabolites. Metabolism leads to many potential compounds (isomers), which complicates the analysis. Thus, to overcome these limitations, non-targeted analysis offers new opportunities for a deeper understanding of metabolites related to steroid metabolism. Molecular networking (MN) appears to be an innovative strategy combining high-resolution mass spectrometry and specific data processing to study metabolic pathways. In the present study, two databases and networks of steroids were constructed to lay the foundations for the implementation of the GNPS-MN approach. Steroids of the same family were grouped together, nandrolone and testosterone were linked to other analogues. This network and associated database were then applied to a few urine samples in order to demonstrate the annotation capacity in steroidome study. The results show that MN strategy could be used to study steroid metabolism and highlight biomarkers.