Building trust in the whitefish value chain: Organisational and technical dimensions of a traceability pilot

2 Book of Abstracts: 8th IMEKOFOODS conference: Food Safety and Traceability for a Sustainable Future 22-24 September 2025, Ljubljana, Slovenia Organised by: Dept. of Environmental Sciences, Jožef Stefan Institute Editors: prof. dr. Nives Ogrinc, dr. Marjeta Mencin Published by: Jožef Stefan Institute Press Issued by: Jožef Stefan Institute For the issuer: prof. dr. Leon Cizelj, v.d. director Cover design: Neža Tomori Kontrec Cover page photo: Canva Pro (licensed image) Ljubljana, 2025 First edition This publication is free of charge. This publication is available in e-book format only: https://conferences.imeko.org/event/12/overview Kataložni zapis o publikaciji (CIP) pripravili v Narodni in univerzitetni knjižnici v Ljubljani COBISS.SI-ID 249075971 ISBN 978-961-264-317-1(PDF) Copyright © 2025 by Jožef Stefan Institute All rights reserved. This book, or parts thereof, may not be reproduced in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system now known or to be invented, without written permission from the publisher. 3 CONTENTS WELCOME 4 SCIENTIFIC ADVISORY BOARD 6 ORGANISING COMMITTEE 6 INVITED SPEAKERS 7 PROGRAMME 11 WORKSHOP: TRACEABILITY AND TRUST SOLUTIONS IN EU FOOD SUPPLY CHAINS 19 ORAL PRESENTATIONS 20 POSTERS 84 4 Welcome Dear readers and delegates, it is a pleasure to present you with this Book of Abstracts consisting of contributions to the 8th IMEKOFOOD conference “Food Safety and Traceability for a Sustainable Future”. The conference organized by Jožef Stefan Institute is scheduled to take place at the Grand Hotel Union in Ljubljana, the capital city of Slovenia. The increasing globalization of food trade, rapid changes in food systems, the pressures of climate change, and the demands of a growing world population are profoundly influencing food safety, quality, and nutrition. These issues are closely tied to food security, malnutrition, and public health outcomes worldwide. Addressing them requires reliable, comparable, and metrologically sound food data that can underpin risk assessment, regulatory decisions, and innovative industry practices. Food is an exceptionally complex biological matrix that contains thousands of organic and inorganic substances. Analytical sciences and food metrology face the ongoing challenge of developing and validating innovative methods that can accurately assess a wide range of chemical contaminants and nutritional components within these matrices. Such efforts are essential to strengthen consumer protection and to ensure confidence in food systems at both national and international levels. In this context, the 8th International IMEKOFOODS Conference (22–24 September 2025) will convene scientists, researchers, and practitioners from diverse fields—including food chemistry and metrology, nutrition, food safety and quality, risk assessment, and food authenticity. The conference will serve as a platform for the exchange of knowledge, the presentation of cutting-edge research, and the establishment of collaborative networks. The program is structured around seven thematic areas, covering: • Novel analytical and metrological approaches for food analysis. • Methods for detecting and quantifying chemical contaminants and residues. • Advances in nutritional analysis and food quality assessment. • Risk assessment strategies supported by reliable measurement data. • Food authenticity, traceability, and fraud detection. • Interdisciplinary approaches linking food safety, nutrition, and public health. • Contributions to sustainable and resilient global food systems. We are especially committed to supporting early-career researchers, encouraging their participation, and fostering the next generation of experts who will carry forward the spirit of IMEKOFOODS. This year’s scientific program includes 8 invited speakers, 61 oral presentations, and 40 poster presentations, featuring contributions from 17 countries across Europe and beyond. The conference opens with two dedicated workshops: • AI in Food Science – Exploring artificial intelligence applications in food research and industry • Soft Skills for Students: Project Writing – A practical guide to writing successful research and project proposals In addition, a Cluster Workshop on the EU Cluster for Food Traceability & Trust will bring together several major EU projects (WATSON, ALIANCE, THEROS, FishEUTrust, Drag4FOOD) to explore innovation, synergies, and collaboration in building trustworthy and transparent food systems. We warmly thank all participants—invited speakers, presenters, moderators, and sponsors—for their valuable contributions. Our sincere gratitude also goes to the organizing and scientific committees, whose dedication has made this conference possible. Dear participants, we hope that the wide range of sessions, workshops, and networking opportunities will inspire you, deepen your knowledge, and strengthen your collaborations. May this conference be an exciting, informative, and rewarding experience for all. We look forward to a fruitful and engaging IMEKOFOODS 2025! Prof. dr. Nives Ogrinc Chair of the IMEKOFOODS Organising committee 5 SPONSORS The organising committee is deeply appreciative of the sponsorship generously provided by the following companies: 6 SCIENTIFIC ADVISORY BOARD Andrea ROSSI: National Institute of Metrological Research (INRiM), Italy - Chair of EMN - Safe and Sustainable Food (EURAMET) Anita NAJDENKOSKA: Institute of Public Health of the Republic of North Macedonia, Republic of North Macedonia Claudia ZOANI: National Agency for New Technologies, Energy and Sustainable Economic Development (ENEA), Italy - Scientific secretary of the IMEKO TC23 Isabel CASTANHEIRA: The National Health Institute Doctor Ricardo Jorge (INSA), Portugal Janaína MARQUES RODRIGUES: National Institute of Metrology, Quality and Technology (INMETRO), Brazil Jens SLOTH: Danish Technical University - National Food Institute (DTU-Food), Denmark Joris VAN LOCO: SCIENSANO, Brussels, Belgium - Chair of the IMEKO TC23 Karl PRESSER: Premotec, Switzerland Lenka KOUŘIMSKÁ: Czech University of Life Sciences, Czech Republic Luc INGENBLEEK: World Health Organization (WHO), Switzerland Maria FERNANDES-WHALEY: National Metrology Institute of South Africa (NMISA), South Africa - Scientific secretary of the IMEKO TC23 Maria TSIMIDOU: Aristotle University of Thessaloniki, Greece Nastasia BELC: National Institute of Research & Development for Food Bioresources, Romania Nives OGRINC: Jožef Stefan Institute (JSI), Ljubljana, Slovenia - IMEKO TC23 Deputy Chairperson - Chair of the 8th IMEKOFOODS conference Petru JITARU: French Agency for Food, Environmental and Occupational Health & Safety (ANSES), France ORGANISING COMMITTEE Nives OGRINC: Conference chair - Jožef Stefan Institute Milena HORVAT: Jožef Stefan Institute Nataša POKLAR ULRIH: Biotechnical Faculty, University of Ljubljana Mojca MILAVEC: National Institute of Biology Milena BUČAR MIKLAVČIČ: ZRS-Koper Helena BAŠA ČESNIK: Agricultural Institute of Slovenia Lidija FRAS ZEMLJIČ: Faculty of Mechanical Engineering, University of Maribor Marjeta MENCIN: Jožef Stefan Institute Cathrine TERRO: Jožef Stefan Institute Anja VEHAR: Jožef Stefan Institute Bor KRAJNC: Jožef Stefan Institute Boštjan KRIŽANEC: National Laboratory for Health, Environment and Food 7 INVITED SPEAKERS Prof. Michele Suman Barilla Analytical Food Science, Parma, Italy & Catholic University of the Sacred Heart, Milan, Italy Conference Title: Versatility of the GC-IMS technique: from the freshness of eggs, to oil fraud, to the origin of apples Prof. Michele Suman is a Food Safety & Authenticity Research specialist with over two decades of experience leading innovative research and development within the agri-food sector, with a track record of method development for studying contaminants, adulterants, authenticity markers and their transformation along food processing, pioneering new technologies as well. He has expertise in coordinating complex, internationally funded projects, and is dedicated to mentoring the next generation of professionals. Through collaboration with private and public stakeholders, he actively fosters innovation and advances the field. Since 2003, Prof. Suman has served as the Food Safety & Authenticity Research Manager in Barilla Spa company. He is also an Adjunct Professor of AgriFood Authenticity at Catholic University Sacred Heart (Milan/Piacenza) and a Visiting Professor at University of Chemistry and Technology in Prague. Additionally, he chairs the Italian National Normative Organization (UNI) Food Authenticity Working Group and contributes as a member of several prestigious committees, including Joint FAO/WHO Expert Committee on Food Additives (JECFA), working groups for Biotoxins-Processing Contaminants-Food Authenticity in European Committee for Standardization (CEN), the ILSI Europe Scientific Advisory Committee, where he also serves as Vice Chair of the Food Contaminants Task Force, the Italian Cluster Agrifood Scientific Board, and the European Food Safety Platform Steering Committee. His scientific impact is reflected in his Google Scholar H-index 32 & i10-index 69, Scopus H-index 28, Research Gate index 30. His scientific production is documented by 6 book chapters, 2 books edited, 180 contributions at national and international conferences and 130 papers in international ISI journals. Dr. Stefan van Leeuwen Wageningen Food Safety Research (WFSR), part of Wageningen University & Research, Wageningen, The Netherlands Conference Title: The analytical toolbox for detection and identification of legacy and emerging PFAS in food production chains Dr. Stefan van Leeuwen is a senior scientist at Wageningen Food Safety Research, with expertise in analytical and environmental chemistry. Driven by a passion for enhancing food safety, he specializes in developing state-of-the-art chromatographymass spectrometry approaches to detect trace levels of environmental pollutants like PFASs, chlorinated and brominated pollutants. Dr. van Leeuwen is also at the forefront of innovation in identifying unknown contaminants, including novel PFAS compounds, by creating a comprehensive analytical toolkit. His current research explores emerging food safety challenges in circular food production, assessing how potential pollutants could pose risks when wasteand by-products are repurposed in food production systems. Through his work, Dr. van Leeuwen is helping to pave the way for a more sustainable and safer future in global food production. He is a co-chair of the biannually organised conference series Recent Advances in Food Analysis. He is co-chair of the Core Working Group on PFASs of the European Reference laboratory on POPs. 8 Prof. Amadeo Rodríguez Fernández-Alba European Union Reference Laboratory for Pesticide Residues in Fruit & Vegetables, University of Almería, Spain Conference Title: New trend in Multiresidue Methods: Achieving the maximum with the minimum Prof. Amadeo is a founder of The Pesticide Residues Group AGR159 created in 1995, recognized as a center of excellence by the Plan Andaluz de Investigación in Spain. He has coordinated 30 national/regional projects, over 60 research contracts with national and international companies, and participated in 16 European projects. His research has produced over 370 ISI-JCR articles (35,000+ citations, h-index 105) and 5 patents, along with 6 edited books, 17 book chapters, and supervision of 28 PhD theses. His work focuses on chemical evaluations using mass spectrometry to identify contamination hotspots, predict trends, optimize remediation technologies, improve legislation, and design control systems. Current research includes microplastic evaluation in the atmosphere using passive samplers on beehives and the reuse of reclaimed water in greenhouse crop production. He also leads studies on balancing food production with minimizing environmental impact, assessing contaminants from pesticides, fertilizers, pharmaceuticals, and plastics, and developing advanced oxidation processes for water and plastic waste treatment. Since 2006, he has been Director of the EU Reference Laboratory for Pesticide Residues in Fruits & Vegetables, contributing to European legislation on pesticide use and control, including the EU’s New Green Deal. Recognizing the laboratory’s contribution to agronomic activities, biodiversity, and ecosystem sustainability, his team has also established a new line of work on the impact of contaminants on declining honeybees populations, participating in Spain’s National Plans for chemical analysis. Prof. María Dolores Hernando Guil CSIC, The Spanish National Research Council, EEZA Experimental Station of Arid Zones Conference Title: Microplastics: A New Hidden Threat in Our Food? Prof. María Dolores Hernando Guil has participated in multiple national and international research projects, including 10 under the Spanish National R&D Plan, 3 European projects and 1 EU-Latin America cooperation project. Her work focuses on environmental contamination, pollutant behavior, and food safety, with emphasis on Persistent, Bioaccumulative, and Toxic (PBT) chemicals, Persistent Organic Pollutants (POPs), emerging contaminants, and microplastics. She has contributed to risk assessments, pollution control strategies, and conservation efforts, particularly in arid regions. She has co-authored 105 JCR journal articles (h-index: 46, 8,000+ citations), 6 book chapters, and the HERWE risk screening tool for chemical pollutants in wastewater effluents. She has also contributed to UN and EU technical reports on chemical risk management and served as Coordinator of Spain’s National Reference Centers for the implementation of the EU REACH Regulation, the Global Harmonized System of Classification, and National Reference Center on Persistent Organic Pollutants (2007–2011). Since 2023, she has been Head of the Department of Desertification and Geo-ecology at the Spanish National Research Council (CSIC). She has participated in more than 150 international conferences, 30 seminars for national business confederations, and supervised 8 PhD theses (with 5 ongoing). She also teaches in Ph.D., Master’s, and technical training programs at several Spanish universities. 9 Dr. Marta Dabrio Ramos European Commission’s Joint Research Centre Conference Title: The Future of Food Analysis: Current Advances and NextGeneration Reference Materials Dr. Dabrio Ramos holds a PhD in Analytical Chemistry from the Autónoma University of Madrid (UAM), Spain, followed by a postdoctoral stay at the Karolinska Institute in Stockholm, Sweden. After a brief period in the private sector, she has been part of the European Commission’s Joint Research Centre for over 20 years. She has worked on the production of Certified Reference Materials, initially as Head of the Laboratory for Small Molecules, and currently serves as Team Leader for Reference Materials for Food Analysis. Séverine Goscinny MSc Department of Chemical and Physical Health Risks, Sciensano, Belgium Conference Title: From concept to laboratory work: challenges and innovation in method validation for food additives. Séverine Goscinny is a Senior Researcher at Sciensano in Brussels, where she has committed 18 years to the Department of Chemical and Physical Health Risks. Her research focuses on the safety and thorough analysis of chemical substances within the food chain, especially flavourings and additives. This involves developing and validating advanced analytical methodologies for their precise detection and accurate quantification in food. Additionally, she carries out chemical analyses to identify and measure various chemicals, which helps refine dietary exposure studies. A key part of her role is risk assessment, where she carefully evaluates the potential risks linked to these chemicals in our food. She is also active in European food safety networks, promoting collaboration and knowledge sharing among colleagues across the continent. Prof. Dr. Michael Rychlik Chair of Analytical Food Chemistry, Technical University of Munich, Germany Conference Title: Stable isotope dilution assays for mycotoxins help assess the risk of plant-based meat alternatives Prof. Dr. Michael Rychlik is heading the Chair of Analytical Food Chemistry at the Technical University of Munich, Germany (TUM). He graduated in food chemistry at the University of Kaiserslautern in 1988. His PhD studies on the flavour of bread were completed in 1996 and he was appointed full professor at the TUM in 2010. In 2015 he served as a Visiting Professor at the University of Queensland (UQ), Australia and in 2016 he was appointed an Honorary Professor at the latter University. In the last years he was also active as a Visiting Professor in 2016 at the National University of Singapore (NUS) and been teaching also in 2018 at the University of Hong Kong. In 2023 he was appointed a Visiting Professor at the University of Chemistry and Technology Prague. His group has been working for 25 years in the field of developing analytical methods for bioactive food components, in particular for vitamins, mycotoxins, odorants and lipids. Since 2014 he serves as the Head of the “Committee on Contaminants in the Food Chain” at the Federal Institute for Risk Assessment, Berlin, Germany. 16 POSTER SESSION, Monday 22. 9. 2025 ADVANCES IN FOOD AND NUTRITION P1 D. A. Alexandra: Antimicrobial profile of sourdough lactic acid bacteria against pathogen bacteria and molds P2 Z. Koziara: The histaminergic system as a potential target in the treatment of binge eating behaviour in an animal model P3 L. Morales Erazo: Recovery and Green Purification of Methylxanthines and Flavan-3-ols from Cocoa Husk Agro-Waste P4 M. Korošec: Dietary Potassium Reduction in Fruits and Vegetables: A Study on the Efficacy of Ultrasonic Soaking and Thermal Processing P5 A. Culetu: Evaluation of side-streams from sea buckthorn oil production P6 A. Culetu: Exploring the amino acid profile of various legumes for optimal nutrition P7 C. Zoani: Co-creation from University-based Living Labs: the case of the FUSTO Programme FOOD AUTHENTICITY & TRACEABILITY P8 C. Terro: Multi-Analytical Assessment of Paprika and Cinnamon Authenticity and Quality in the Slovenian Market P9 Z. Sel: Analytical Characterisation of Emerging High-Value Seed Oils in Slovenia P10 M. Mencin: Impact of Climate Change on Fatty Acid Composition and δ¹³C Signatures in Slovenian Extra Virgin Olive Oil: A Long-Term Study P11 N. Poklar Ulrih: Quality Assessment and Adulteration Detection of Rosemary and Laurel Essential Oils Using GC-C-IRMS P12 G. Puzo: Integrated Multi-Method Approaches to ensure Olive Oil Authenticity and Traceability P13 A. Bernardini: Territorial origin determination of wheat flour by NMR profiling: a case study P14 L. Kpattah: Tracing the Sources of Adulteration along the Ghanaian Food Supply Chain using Isotopic and Chromatographic Techniques: Implications for Food Authenticity FOOD DATA ANALYSIS P15 G. Mustatea: Al-Driven Review of Analytical Methods for Microand Nanoplastics Analysis P16 M. Zampano: Real-time AI-based monitoring system for microbiological safety of drinking water METROLOGY IN FOOD P17 M. Z. Tsimidou: METROFOOD-RI integrated analytical services: Phenol determination in agroindustrial by-products and derived formulations P18 M. Khayrullaev: The Impact of Equipment Conformity Assessment in Ensuring the validity of Food Testing: A Comparison of Laboratory Practices P19 C. Vansnick: Metal impurities in food additives: Do they contribute to dietary metal exposure? P20 A. Bogožalec Košir: Standardizing Plant Pathogen Detection using dPCR as a Reference Method for Xylella fastidiosa P21 P. Marinček: Development and validation of control materials for molecular detection of Listeria monocytogenes in milk P22 O. Shcherbakova: Grain moisture determination. Using of standard addition method P23 D. Chupis: Metrology for the phosphorus content measurements in vegetable oils in support of European and international food safety 17 POSTER SESSION, Tuesday 23. 9. 2025 FOOD PACKAGING P1 M. Di Mario: What is the future of FCM after the ban on single-use plastic articles? A deep dive into alternative materials in Belgium. P2 F. Romaniello: Enhanced untargeted method to detect organic and inorganic contaminants in biobased and recycled food contact materials P3 T. Šaula: Microplastics on the Surface of Packaged Chicken Meat: Method Optimization for Detection, Isolation, and Characterization P4 A. John: Poly (lactic acid)–Poly (L-lactide-co-ethylene adipate) Hybrid Films: A Promising Approach for Sustainable Food Packaging FOOD PROCESSING P5 L. Kourimska: Food Processing Workshops in the RURBANIVE Project: Helping Local Producers Shorten the Supply Chain P6 T. Šaula: Antibacterial Potency of Slovenian Honeys against Foodborne Pathogens P7 I. Petrignani: Rapid and non-destructive assessment of cultivars and geographical origin of durum wheat flour using Raman spectroscopy NANOPARTICLES IN FOODSTUFFS P8 U. Rozman: Microplastics in food: literature review P9 N. Renier: Identifying Technological Needs in Microand Nanoplastics Research: A FHERITALE Perspective ANALYTICAL APPROACHES IN FOOD CONTAMINANTS P10 H. Baša Česnik: Multiresidual GC-MS/MS method for determination of pesticide residues in fruits P11 H. Baša Česnik: Multiresidual GC-MS/MS method for determination of pesticide residues in vegetables P12 M. L. Adriana: GC-MS/MS method for acrylamide determination in various food matrices from Romania P13 M. L. Adriana: A quenchers-based protocol for analysis of polycyclic aromatic hydrocarbons (PAHs) in vegetable oils and their derivatives P14 M. L. Adriana: Validation study according to commission regulation (EU) 2021/808 for determining PAHs in cereal derivatives by GC-MS/MS P15 T. Radovanović Vukajlović: Fungal and Yeast Contaminants in Dried Fruits: Implications for Food Quality and Safety P16 I. Bastardo-Fernandez: High content, low release: Exploring TiO2 nanoparticle migration from food packaging using APEX™-single particle ICP-MS/MS P17 V. Alampanos: Trace-level determination of PFAS in water samples using LC-HRMS and a novel MOF sorbent in miniaturized SPE 18 CLUSTERING P18 M. Pirc: Consumer Perspectives on Food Fraud Vulnerability and Traceability: Insights from a Cross-Country Survey P19 L. Karapetsi: Galician mussel species identification via HRM analysis and ML for automated DNA Classification P20 D. Kocman: User-centred pilot testing of tools for organic and geographical indication of food products P21 Lorenzo Nolfi: Biosafety assessment of raw materials and soil improvers for a safe use in agriculture 19 The EU Cluster for Food Traceability and Trust is a collaboration of currently 13 European research and innovation projects that share a common ambition: to make food systems more transparent, trustworthy, and resilient. The cluster brings together initiatives exploring how digital technologies such as blockchain and artificial intelligence, combined with improved organisational practices, can help address longstanding challenges of food fraud, safety, and consumer confidence. By joining forces, the projects seek not only to advance technical solutions but also to open dialogue across disciplines and stakeholders — from producers and retailers to policymakers and consumers — about what it truly takes to build trust in the food we eat. This workshop offers participants a chance to hear about emerging solutions from across the cluster and to contribute to shaping their future direction. The morning begins with short research presentations on traceability pilots in the whitefish value chain, consumer attitudes towards blockchain-tracked olive oil and feta cheese, and a blockchain-based supply chain model for PDO feta. After a short break, the session shifts into a more interactive mode: solution “snapshots” from additional projects, breakout discussions on barriers and opportunities, and a plenary exchange to distil key insights. The discussions will focus on real-world questions: What prevents blockchain and AI from being widely deployed in food systems? What kinds of evidence and data are needed to inspire trust among consumers and stakeholders? How can EU projects better connect their tools, knowledge, and datasets? And what kinds of policy frameworks or market incentives are missing for scaling these innovations? Importantly, this workshop will bring together participants from WATSON, ALLIANCE, THEROS, FishEUTrust, and Drag4Food, ensuring a rich mix of perspectives from across the EU’s food traceability innovation landscape. Rather than producing only formal conclusions, the session is designed to generate collaborative outputs — ranging from shared perspectives and knowledge gaps to a joint workshop summary, a cocreated brainstorming space, and even a short blog post co-authored with participants. By the end, we hope to leave not only with sharper questions but also with stronger connections across projects and stakeholders who care about trustworthy food supply chains in Europe. Abstract presentations: K. Jakobsen (WATSON): Building trust in the whitefish value chain: Organisational and technical dimensions of a traceability pilot S. Sadrmousavigargari (ALLIANCE): Consumer Purchase Intentions for Blockchain-Tracked Olive Oil and Feta Cheese K. Choumas (ALLIANCE): Blockchain-based supply chain for PDO Feta Cheese: The Alliance approach P. Lappas (ALLIANCE): Securing Agri-Food Supply Chains with AI for Early Warning, Fraud Detection, and Decision Support M. Pirc (WATSON): Consumer Perspectives on Food Fraud Vulnerability and Traceability: Insights from a Cross-Country Survey L. Karapetsi (THEROS): Galician Mussel Species Identification via HRM Analysis and ML for Automated DNA Classification D. Kocman (THEROS): User-centred Pilot Testing of Tools for Organic and Geographical Indication of Food Products Workshop: Traceability and Trust Solutions in EU Food Supply Chains 20 ORAL PRESENATIONS 21 Versatility of the GC-IMS technique: from the freshness of eggs, to oil fraud, to the origin of apples Michele Suman Catholic University of Sacred Heart Milan-Piacenza, Italy This presentation explores the versatility of Gas Chromatography–Ion Mobility Spectrometry (GCIMS) as a rapid, sensitive, and cost-effective analytical tool for food quality control. Three case studies demonstrate its application across diverse food matrices: egg freshness, geographical origin of dehydrated apples, and authenticity of extra virgin olive oil (EVOO). In the first case, GC-IMS was employed to assess the freshness of egg products under various storage conditions. Identifying several volatile markers, the method proved more sensitive than official protocols and suitable for 100?tch screening in production environments. The second case focused on verifying the “100% Italiano” claim for dehydrated apples used in food industry. A comprehensive experimental design considered variables such as variety, peeling, dehydration, and harvest year. GC-IMS data, processed through PCA and OPLSDA, enabled clear discrimination between Italian and non-Italian origins and the model achieved more than 90?curacy in validation sets. The third case addressed EVOO fraud detection, particularly the undeclared blending with soft-refined oils (SROOs). GC-IMS fingerprinting revealed distinct VOC profiles between authentic EVOOs and adulterated samples. SIMCA analyses confirmed the method’s ability to detect subtle changes due to soft deodorization and deacidification. The technique demonstrated high repeatability and stability, making it ideal for industrial screening. Overall, GC-IMS offers a powerful, highthroughput solution for food authenticity and quality assurance, with minimal sample preparation and strong potential for broader implementation in quality control laboratories. 22 The Future of Food Analysis: Current Advances and Next-Generation Reference Materials Marta Dabrio European Commission, Joint Research Centre, Belgium The food systems in Europe are complex and vulnerable to challenges such as climate change, food security and geopolitical tensions. In this context, to maintain the resilience and sustainability of the food system, the European Union’s food legislation plays a crucial role in ensuring the safety and authenticity of the food supply chain. The EU’s strict food safety and authenticity guidelines, such as those related to labelling, tracing and testing, are designed to protect consumers from contaminants, foodborne illnesses and deceptive practices, and to maintain trust in the food system. The food industry and food authorities control the food safety, quality, and authenticity by relevant food testing methods. The role of reference materials in ensuring the accuracy and reliability of food analysis has never been more critical. This presentation will explore the vital role of reference materials in modern food analysis, with some examples, highlighting their importance in ensuring the safety, quality, and authenticity of food products. The talk will cover the current state of reference materials in food analysis, including recent advances in their production, characterization, and application. The presentation will furthermore discuss the challenges and opportunities in the field, including the need for reliable reference materials in emerging areas, the development of new technologies and methods, and the importance of international collaborations and standardization efforts. 23 EU policy on contaminants in food: Recent developments, outlook and challenges Frans Verstraete European Commission, Belgium The EU legislation on contaminants Council Regulation (EEC) No 315/93 of 8 February 1993 provides that food containing a contaminant in an amount which is unacceptable from the public health viewpoint shall not be placed on the market (food can only be placed on the market when it is safe). Furthermore, it is foreseen that contaminant levels shall be kept as low as can reasonably be achieved by following good practices at all stages of the production chain and in order to protect public health, maximum levels for specific contaminants shall be established where necessary. In recent years, regulatory levels for several contaminants have been established and updated and this to take account of the outcome of risk assessments performed by the European Food Safety Authority (EFSA). The maximum levels are established in Commission Regulation (EU) 2023/915 of 25 April 2023 on maximum levels for certain contaminants in food and repealing Regulation (EC) No 1881/2006 (http://data.europa.eu/eli/ reg/2023/915/2025-01-01). Despite frequent updates to the EU contaminants legislation, there is still a lot of work ahead! In the presentation, recent and future developments on EU legislation on contaminants in food shall be presented. Climate change, changes in dietary patterns, novel/new foods, circular economy etc. entail new challenges for the safety of the food chain. In addition, in order to continue to ensure a high level of food safety it is necessary not to address single contaminants individually but also pay more attention to the combined exposure to multiple contaminants. In addition, particular attention shall be paid to the analytical requirements and analytical challenges for an effective EU policy on contaminants in food. Indeed, for an effective risk management and enforcement, it is not only sufficient that a method of analysis is available, but the method of analysis must be able to be used for routine control, be reliable, sensitive, quick and preferably cheap. 24 Building a quality characteristics database in support of the geographical indications for the Greek saffron ˝Krokos Kozanis˝ Maria Z. Tsimidou, Stella Ordoudi Aristotle University of Thessaloniki - AUTH , Greece Saffron is the spice that comes from the drying of the red stigmas of Crocus sativus L. flower. This most expensive spice by weight in the world, is of great importance for the local community around the village Krokos (Kozani, Western Macedonia, Greece). This work presents the results of a systematic study on the compositional variability in Greek saffron in terms of three determinant criteria defined by international trade standards. A total of 547 samples (2022 harvest) were obtained from registered growers - members of the Cooperative of Kozani Saffron Growers. The administration and quality control managers of the Cooperative coordinated sample collection and direct shipment to our laboratory for analysis according to ISO 3632-2 test methods to assess moisture content, and also coloring, flavor and aroma strength indices. The analytical data were combined with metadata about individual member production quantity, processing practices, etc that helped overall interpretation and discussion. The results of the systematic study were used to start building a compositional database for the Protected Designation of Origin saffron “Krokos Kozanis”. This database needs regular updating to serve not only the internal quality control management system of the Cooperative but also to provide robust documentation for official controls in support of the registered Geographical Indication that enhances consumer confidence in fair trade practices. Acknowledgement: Part of this research is linked to the project ELKE/AUTH contract no 278709, financed by the Kozani Saffron Growers Cooperative. The project ‘Dissemination and exploitation of the outcomes of the Laboratory of Food Chemistry and Technology’ (AUTh Research Committee code 96677) is acknowledged for financial support of MZT participation. FOOD AUTHENTICITY AND TRACEABILITY 25 Isotopic fingerprinting of PDO tomatoes: a tool for authentication and traceability Raffaela Ofano1, Doris Potočnik2, Nives Ogrinc2, Paola Adamo1 1Department of Agricultural Sciences, University of Naples Federico II, Italy 2Jožef Stefan Institute, Slovenia The authentication and geographical traceability of agri-food products are essential strategies to ensure food safety and quality. These approaches not only safeguard the integrity of Protected Designation of Origin (PDO) products but also enhance their market competitiveness and prevent fraud. Soil, as the primary source of mineral elements for plants, influences the elemental and isotopic profile of agricultural products based on bioavailability and pedoclimatic conditions. In this study, stable isotope ratio analysis was employed to investigate the geographic provenance of soils and PDO tomatoes grown on the slopes of the Somma-Vesuvius volcanic complex, a small geographical area where unique pedoclimatic conditions influence their quality and organoleptic properties. Soil and tomato samples were collected from representative farms located both within and outside the PDO-designated area over four years (2021-2024). The isotopic ratios δ34S, δ15N, δ13C and S, N, C concentrations were measured by Isotope Ratio Mass Spectrometry (IRMS) in both tomato and soil samples. The resulting data were subjected to supervised multivariate discriminant analysis, specifically Linear Discriminant Analysis (LDA), to evaluate the discriminatory power of the isotopic parameters for geographic origin authentication. Statistical validation using leave-one-out cross-validation revealed that the highest classification accuracy was achieved when data from each year were analyzed independently, rather than as a combined dataset. Among the isotopic parameters considered, δ34S was the most discriminant markers across all individual years and in the combined dataset. The strong correlation observed between the isotopic profile of tomatoes and their corresponding soils confirms the robustness of this approach. These findings support the use of isotope ratio analysis as a reliable and effective analytical tool for food traceability, contributing to the protection and valorization of PDO products. Research granted by METROFOOD-IT project [code IR0000033] funded by the NextGenerationEU under the National Recovery and Resilience Plan (NRRP), M4C2 Investment 3.1. FOOD AUTHENTICITY AND TRACEABILITY 32 microbiological indicators in fostering resilient food systems and supporting a robust ‘farm to fork’ strategy. Funded by DELISOIL Horizon Europe GA No. 101112855, SIMBA Horizon 2020 GA No. 818431, “Research and innovation network on food and nutrition Sustainability, Safety and Security—Working ON Foods” PE00000003, SUS-MIRRI.IT” IR0000005, SOIL-HUB2 D.M. MASAF n 672924 del 20-12-24. METROLOGY IN FOOD 33 Nutritional and Sensory Impact of Cricket and Mealworm Meals Inclusion in Sourdough Bread Nika Samsa, Jasna Bertoncelj, Blaž Ferjančič, Tomaž Požrl, Mojca Korošec University of Ljubljana, Biotechnical Faculty, Slovenia Edible insects are emerging as a sustainable food source, recognized for their high-quality proteins, fats, minerals, and bioactive compounds, offering a viable strategy for protein diversification. Despite their nutritional potential, regulatory status as novel foods (EU) and consumer acceptance remain significant challenges. This study investigated the nutritional and sensory implications of incorporating ground dry cricket (Acheta domesticus) and mealworm (Tenebrio molitor) meals into sourdough bread. Wheat flour was partially substituted at 5% and 10% (w/w). Chemical analyses revealed that insect inclusion significantly increased bread protein content by 10-25%, dietary fibre by approximately 2%, and ash by 2-8%, while concomitantly reducing available carbohydrates by up to 7%. Notably, the 10% mealworm meal substitution nearly doubled the bread’s fat content. A professional sensory panel observed that higher insect meal concentrations (10%) negatively impacted bread characteristics, leading to uneven crumb structure, irregular loaf shape, gritty texture, and undesirable earthy/animallike flavours, particularly in cricket-enriched samples. Conversely, breads with 5% insect meal exhibited more favourable sensory profiles, though slightly inferior to the 100% wheat flour control. These findings indicate that a moderate inclusion (5%) of ground mealworms and crickets can effectively enhance the nutritional value of sourdough bread without severely compromising sensory quality, representing a promising step toward developing sustainable and nutritionally superior food products. However, the intrinsic aroma of insect meals within the bread matrix necessitates further research into consumer acceptability. METROLOGY IN FOOD 34 WDXRF as a tool for the characterisation of a candidate MRC for major and minor elements in rice flour Linda Alejandra Pérez Díaz1, Juliana Serna Saiz1, María José Martinez Cordon2 1Instituto Nacional de Metrología , Colombia 2Universidad Nacional de Colombia, Colombia The development of a Certified Reference Material (CRM) for elemental analysis of rice is essential to ensure food quality and safety. Currently, few National Metrology Institutes (NMIs) have CRMs for major and minor elements in the rice matrix, with the most recognized being SRM 1568b from the U.S. National Institute of Standards and Technology (NIST). This highlighted the need to produce a national CRM to improve accessibility within the country. Additionally, only a limited number of national laboratories are accredited under the NTC/ISO 17025:2017 standard for measuring elements in rice, underscoring the importance of promoting accreditation to guarantee reliable results. Commonly used analytical techniques—atomic absorption, ICP-MS, and ICP-OES—are accurate but costly and destructive. In contrast, X-ray fluorescence (XRF) is a more economical, non-destructive technique with minimal sample preparation, but it remains underutilized due to the lack of accredited laboratories. To address this situation, a candidate rice flour CRM was prepared and characterized using wavelength-dispersive X-ray fluorescence spectrometry (WDXRF) for the simultaneous determination of major (Mg, Ca, Na, P, S, K) and minor (Mn, Zn, Cu, Fe) elements. Calibration curves were established using NIST-certified reference materials of organic matrices, while other CRMs were used to validate the method. Samples were dried according to the conditions stated in each CRM certificate and prepared as pressed pellets (1 g sample with 0.11 g wax, pressed at 20 tons for 120 seconds). The developed method proved to be selective, precise (with relative standard deviations below the Horwitz coefficient), and accurate, as biases were within the limits defined by the standard uncertainty of each CRM and the experimental standard deviation. Therefore, the observed bias was considered statistically insignificant. Final characterization of the reference material INM-027-1 by WDXRF yielded the following mass fraction values: Ca (85.30 mg/kg), Cu (3.21), Fe (13.37), K (2875.55), Mg (1672.51), Mn (28.78), P (3751.75), Zn (20.89), and S (1275.26), with relative uncertainties ranging from 0.2% to 19.4%, depending on the element. METROLOGY IN FOOD 35 Revealing metabolic alterations in plants induced by nanoand microplastic exposure by high-resolution mass spectrometry María Dolores Hernando Guil1, María José Gómez-Ramos2, Harshit Sahai2, Amadeo R. Fernández-Alba2 1Experimental Station of Arid Zones, CSIC, Spain 2University of Almeria, Spain This study represents a novel comprehensive assessment of metabolic perturbations elicited by nano and microplastics (NMPs) in plant systems, employing Lactuca sativa as a model organism. It was accomplished through controlled hydroponic exposure assays with high resolution mass spectrometry. It was found that plastic nanoparticles disrupt metabolic pathways by altering levels and activities of polyunsaturated fatty acids (PUFAs), revealing a previously unrecognized interaction between NMPs and plant biochemistry. Particle size proved critical: the smallest nanoparticles (100 nm and 200 nm) produced the most pronounced metabolic disturbances, with 100 nm particles causing the greatest shifts in PUFA-associated molecular masses, as evidenced by significant changes in chromatographic peak areas. Specifically, the molecular ion at m/z 293.2469—tentatively identified as the methanol adduct of glycerolipid (6Z,9Z,12Z,15Z)-octadeca-6,9,12,15-tetraenal—was most significantly affected at 100 nm. Two additional ions, m/z 307.2264 and 309.2415, also exhibited heightened sensitivity to smaller particles, underscoring the size-dependent nature of these interactions. By contrast, the methanol adduct of α-linolenic acid (m/z 311.2575) responded more gradually across particle sizes, suggesting a distinct interaction mechanism. Larger particles (500 nm and 1000 nm) had markedly weaker effects, confirming that smaller nanoparticles are the primary drivers of PUFA downregulation. These findings demonstrate the power of precise-mass metabolomics for reliable data generation when assessing NMPs impacts and provide crucial insights into their potential implications for plant health and food quality. 36 The Knowns And The Unknowns: The Analytical Toolbox To Tackle The PFAS Challenge Stefan van Leeuwen Wageningen Food Safety Research, Wageningen University & Research (WUR), Netherlands PFAS pollution has evolved to an unprecedented societal challenge. PFASs are found everywhere in our environment and foods, the number of compounds is continuously expanding, and already at low levels exposure may be exceeding the safe levels. Food and drinking water are important vectors for human exposure, and a diversity of analytical tools are needed to unravel the PFAS problem in food. The toolbox consists of 3 elements, that allow the investigation of the following: - Mass balance analysis, i.e. the determination of the total PFAS amount in foods - Targeted analysis, i.e. the (sub) pg/g level determination of PFASs in foods and feeds - Unknowns identification, i.e. the screening and identification of (yet) unknown PFASs in foods. Highly sensitive targeted approaches for reliable, ultrasensitive (low/ sub pg/g level) are developed for carboxylic acids, sulfonates, telomer sulfonates, telomer alcohols, sulfonamides, ultrashort-chain PFAS etc. This provides robust data for the purpose of e.g. monitoring, compliance testing and exposure assessment. The mass balance approach we applied on freshwater fish samples, showing that over 50% of the extracted organic fluorine could not be attributed to the know PFASs (as determined by targeted approaches). This means that a substantial amount of yet unknown PFASs are present in these samples. A Thermo Scientific Orbitrap IQ-X Tribrid HRMS is used to identify the unknown PFASs causing this missing proportion of extracted organic fluorine. For that purpose, we have developed a workflow to screen for suspects in sample extracts. In addition, advanced acquisition modes were developed combined with specific data filtering tools (i.e. Kaufman approach, Kedrick mass defect) are applied to elucidate unknown PFAS signals in food extracts, which the need further confirmation by (ideally) Level 1 identification using reference standards. Altogether, this toolbox allows for unravelling the PFAS complexity in foods and environmental samples, as it provides insights into PFASs contamination from different angles. Next to current state-of-the-art, analytical challenges will be addressed, as well as the needs to advance this field further in the future. 37 Data fusion of multi-source spectral data for the authentication of food and beverages Ariana Raluca Hategan, Maria David, Dana Alina Magdas National Institute for Research and Development of Isotopic and Molecular Technologies, Cluj-Napoca, Romania Owing to their high market value, wine and honey represent agri-food products that are frequently subjected to fraud, particularly in regard to their geographical and botanical origin, as well as harvesting year. As conventional authentication techniques such as stable isotope ratio analysis often present limitations in terms of costs, destructive nature, or sample preparation and analysis time, there has been growing interest in employing spectroscopic methods coupled with advanced data processing tools for the development of reliable recognition models. More recently, the integration of multiple spectroscopic data sources through data fusion approaches has emerged as a powerful approach for improving the accuracy and reliability of authenticity assessments. Against this background, the present study explores the application of data fusion strategies combining 1H NMR and Raman spectroscopies for wine and two complementary vibrational techniques, Attenuated Total Reflection Fourier Transform Infrared (ATR FT-IR) and Fourier Transform Raman (FT-Raman) spectroscopies, for honey recognition. For wine, a dataset of more than 50 authentic white wine samples from Romania, encompassing four cultivars across three viticultural regions over five vintages, was analyzed. Given the high dimensionality of the fused spectral data, a supervised feature selection based on Partial Least Squares (PLS) regression was applied to identify the most discriminative markers. The resulting models demonstrated outstanding classification performances, achieving up to 100?curacy in both cross-validation and external testing, with the fused data outperforming single-technique inputs, especially in varietal discrimination. Similarly, in the case of honey authentication, low-level data fusion coupled with a subsequent supervised feature selection step allowed for enhanced recognition of honey botanical origin and harvesting year. The obtained results underscore the potential given by the association of multi-source spectroscopic data for the authentication of complex food matrices such as wine and honey, which not only enhances model performance but also contributes to the construction of more comprehensive and reliable food and beverages authentication models. Acknowledgements: This paper was co-financed by the European Regional Development Fund (ERDF) through the Smart Growth, Digitization, and Financial Instruments Program (PoCIDIF), call PCIDIF/144/ PCIDIF_P1/OP1/RSO1.1/PCIDIF_A3, Project SMIS number 309287, acronym METROFOOD-RO Evolve. FOOD AUTHENTICITY AND TRACEABILITY 38 Integrating Elemental and Isotopic Analysis for Wheat Authentication and Traceability Giulia Puzo1, Emilia Pucci2, Valeria Poscente2, Alessandra Bernardini2, Filippo Sevi2, Doris Potočnik3, Nives Ogrinc3, Tea Zuliani3, Massimo Reverberi1, Claudia Zoani2 1Sapienza University of Rome, Italy 2ENEA, Casaccia Research Centre, Department for Sustainability, Italy 3Institute Jožef Stefan, Slovenia Wheat is a strategic crop in the global food system, as it is one of the most widely cultivated crops worldwide and serves as a primary source of calories and nutrients for a large portion of the global population. Due to its economic and nutritional importance, ensuring the authenticity and traceability of wheat throughout the supply chain is becoming increasingly important, not only for food safety and consumer protection, but also to prevent fraud, ensure product quality, and promote sustainable agricultural practices [1]. Moreover, in a context of growing global trade and complex supply chains, the ability to verify the geographical origin and production practices of wheat is essential to support fair competition, reinforce labeling systems, and comply with international food standards. This study aims to evaluate the application of integrated chemical-analytical techniques for the discrimination and authentication of wheat samples, emphasizing their potential to enhance quality, safety, traceability, and transparency across the entire supply chain. Samples of Triticum turgidum subsp. durum (durum wheat) and Triticum aestivum L. (soft wheat) were characterised for mineral nutrient composition, the presence of toxic and potentially toxic elements, and stable isotope ratios. In parallel, stable isotope ratio analyses were conducted to investigate variations in isotopic composition associated with the chemical-physical fractionation of elements. The results suggest that integrating multiple chemical-analytical techniques is essential to establish a comprehensive profile in terms of authenticity and geographic traceability. This combined approach provides detailed insights into nutrients and contaminant levels, as well as natural isotopic signatures, ultimately contributing to greater transparency, increased consumer trust, and the valorisation of high-quality wheat products in global markets. Acknowledgment: This work was funded by AGRITECH “National Research Centre for Agricultural Technologies” which received funding from the European Union – NextGenerationEU, under the Italian National Recovery and Resilience Plan (NRRP) – Mission 4 “Education and Research”, Component 2 “From Research to Business”, Investment 1.4: Strengthening research structures and supporting the creation of “national R&D leaders” on key enabling technologies —CN00000022 (D.D. 1032 17/06/2022) This work was supported by METROFOOD-IT, which received funding from the European Union - NextGenerationEU, under the Italian National Recovery and Resilience Plan (NRRP) - Mission 4 “Education and Research”, Component 2 “From Research to Business”, Investment 3.1: Fund for the realisation of an integrated system of research and innovation infrastructures—IR0000033 (MD Prot. n. 120 of June 21, 2022). References: [1] Food and Agriculture Organization of the United Nations, Cereal Supply and Demand Brief, FAO, Rome (2021), available at https://www.fao.org/worldfoodsituation/csdb/en/ FOOD AUTHENTICITY AND TRACEABILITY 39 AGRITECH Project: integrated methodologies for geographical and varietal traceability of agricultural and food products Claudio Rossi, Gabriella Tamasi, Giampiero Cai, Marco Romi, Cristiana Tozzi, Angelo Riccaboni University of Siena, Italy Geographical and varietal traceability of food products plays a crucial role in ensuring their authenticity, quality, and safety—both for agricultural raw materials and final food products. In the global food market, consumers and producers alike are increasingly interested in knowing the geographical origin and the variety of products for purposes of valorization and authentication. This is especially important for high-value products such as wines and extra-virgin olive oils, but also cereals, fruits, vegetables, and dairy products, where origin and variety can significantly influence quality and market value. To achieve reliable and accurate traceability, we employed a combined approach that integrates physical/ chemical and genetic analyses. Advanced experimental spectroscopic and spectrometric methods are applied, including hyphenated chromatographic/mass spectrometry, fluorescence spectroscopy, nuclear magnetic resonance (NMR), and electron paramagnetic resonance (EPR). These cutting-edge tools analyze the elemental, isotopic, and metabolomic characteristics of samples, providing specific fingerprints that can be linked to particular geographic regions or varieties. Genetic analysis also plays a key role in traceability by identifying different varieties within the same species and tracking a specific variety throughout the food production chain. This is achieved using molecular microsatellite markers, which differentiate genotypes based on the number of repeat sequences. These data serve as indicators, helping to verify the authenticity of products and distinguish genuine items from counterfeits or adulterated ones. The data obtained from multiple techniques are then integrated through multivariate statistical methods, including data fusion approaches. This integration enables the development of models capable of interpreting complex datasets and extracting meaningful patterns and correlations. These comprehensive approaches represent a significant advancement in food authentication and valorization, providing stronger tools for consumer protection, quality certification, and the fight against food fraud. This study was carried out within the Agritech National Research Center and received funding from the European Union Next-GenerationEU (PIANO NAZIONALE DI RIPRESA E RESILIENZA (PNRR) – MISSIONE 4 COMPONENTE 2, INVESTIMENTO 1.4 – D.D. 1032 17/06/2022, CN00000022). This manuscript reflects only the authors’ views and opinions, neither the European Union nor the European Commission can be considered responsible for them. FOOD AUTHENTICITY AND TRACEABILITY 40 Nutritional Value of Edible Insects as Alternative Food and Feed Protein Sources in the Czech Republic Lenka Kourimska1, Anezka Kopecka1, Martin Kulma1, Katerina Sebelova2, Jana Hajslova2 1Czech University of Life Sciences Prague, Czech Republic 2University of Chemistry and Technology Prague, Czech Republic The nutritional value of edible insects depends on many factors, including insect species, rearing conditions, and processing methods—including the way of killing. This study aimed to comprehensively monitor key nutritional parameters of six insect species collected from 13 rearing facilities in the Czech Republic. A total of six sampling rounds were conducted at approximately quarterly intervals to cover seasonal variation (2023: March, July, November; 2024: February, May, November). Altogether, 83 insect samples with a total fresh weight of 28.3 kg were obtained. The analysed species (Tenebrio molitor, Zophobas morio, Acheta domesticus, Alphitobius diaperinus, Gryllus assimilis, and Blaptica dubia) were killed either by freezing (-18 °C) or by blanching in boiling water for 5 minutes. In lyophilized samples, the following parameters were determined: dry matter and ash content (gravimetrically), total lipids (Soxhlet extraction), and crude protein content (Kjeldahl method with a nitrogen-to-protein conversion factor of 6.25). Nutritional values varied most significantly by insect species. Observed ranges were dry matter 16.3–41.0 g/100 g, ash 0.59–6.44 g/100 g of fresh weight (FW), fat 1.6–16.9 g/100 g FW, and crude protein 10.0–26.3 g/100 g FW. The way of killing had no significant effect on nutritional composition. However, significant differences in lipid, protein, and ash content were observed within individual insect species, depending on the season and rearing facility (rearing conditions including feed). Therefore, when using insects as food or feed, it is essential to assess their specific nutritional profile from each supplier. Additionally, the way of killing should also be recorded and considered. Acknowledgments: The study was supported by the METROFOOD-CZ research infrastructure project (MEYS grant number: LM2023064), including access to its facilities, and by the Ministry of Agriculture NAZV, QK23020101. ADVANCES IN FOOD AND NUTRITION 41 Comprehensive evaluation of the nutritional composition of chub mackerel (Scomber colias): Influence of cooking methods Andreia Rego1, Fernanda Vilarinho1, Filipa Matias1, José Armando Silva2, Isabel Castanheira3, Carla Motta1 1Instituto Nacional de Saúde Doutor Ricardo Jorge, Portugal 2Instituto Superior Técnico da Universidade de Lisboa, Portugal 3Centro de Ciências do Mar e do Ambiente, FCT-UNL, Portugal Aim: Chub mackerel (Scomber colias) is an abundant but underutilized fish species in the Atlantic, increasingly recognized for its nutritional benefits, including significant contents of omega-3 fatty acids (EPA and DHA). However, comprehensive data on its nutritional profile, especially the impact of cooking methods on nutrient retention, remains scarce. This study aimed to evaluate the nutritional composition of chub mackerel, focusing on moisture, total fat, protein, ash, fatty acid profiles, vitamin B12, and amino acids (AA), comparing raw versus cooked (boiled and grilled) samples. Methods: Chub mackerel from Portuguese Atlantic waters were sampled, and fish dimensions and weights were recorded. A subset was cooked by boiling and grilling. Nutritional analyses included moisture (oven drying to constant weight), ash (incineration at 525 °C), protein (Kjeldahl), and total fat (acid hydrolysis and extraction). The fatty acid profile was assessed by gas chromatography. Additionally, vitamin B12 was analyzed using ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry, and AA were analyzed via ultra-performance liquid chromatography with photodiode array detection. Results: The protein content remained consistently high across all preparations. It also presents all essential AA, each with a score above 1.0, indicating a protein of high biological value. The total fat content varied, with lower levels observed in grilled samples due to fat loss during the cooking process. Total mineral content was relatively stable. The fatty acid profile revealed significant amounts of EPA and DHA. Calculating the retention factor after cooking reveals that the fatty acid content does not vary with processing. Vitamin B12 content increased in cooked samples, from 27.8 µg/100 g in raw to 35.8 µg/100 g (boiled) and 38.1 µg/100 g (grilled). Consuming 100 g of chub mackerel provides more than 100% of the recommended daily intake. Discussion: The results highlight the nutritional robustness of chub mackerel across various culinary methods, particularly in terms of protein and omega-3 fatty acids. The apparent increase in vitamin B12 content after cooking is likely due to water loss, concentrating the nutrient, and enhanced extraction efficiency resulting from protein denaturation. Most AA, particularly essential ones, remain stable during moderate cooking processes. These findings highlight the significance of this species as a valuable source of high-quality nutrients. Conclusion: Chub mackerel is a nutritionally rich and resilient food source, maintaining high-quality nutrient profiles through various cooking processes. Its exceptional content of vitamin B12 and omega-3 fatty acids supports its inclusion in strategies for improving dietary quality and promoting sustainable nutrition. ADVANCES IN FOOD AND NUTRITION 48 Elemental and Metabolomic Characterization of Coffee Plant By-Products for Functional Beverage Development Filippo Sevi1, Alessandra Bernardini1, Giulia Puzo2, Valeria Poscente1, Matteo Nava3, Massimo Battaglia4, Gianfranco Diretto1, Claudia Zoani1 1Italian National Agency for New Technologies, Energy and Sustainable Economic Development (ENEA), Italy 2Università Sapienza, Rome, Italy 3University of Tuscia, Viterbo and ENEA, Italy 4Accademia del Caffè Espresso, Fiesole (FI), Italy Coffee is one of the most widely consumed beverages worldwide. Currently, only the beans are commercially exploited in the food industry, making them the sole source of income for coffee growers. This limited use is critical in terms of sustainability, as the entire coffee production chain is associated with a significant environmental footprint, largely due to the intensive use of water, land, fertilizers, pesticides, and fossil energy. The production process also generates substantial amounts of waste: it is estimated that approximately 0.9 kg of waste is produced for every kilogram of harvested coffee cherries. Among the various by-products are residues from plant pruning and the extraction of bean from the berry. In this context, the valorization of coffee by-products within a circular economy framework is essential for reducing environmental impact. Consequently, in the present study, through chemical and metabolomic approaches, freeze-dried leaves and fruit skins derived from two Caturra varieties (Rojo and Amarillo) of Coffea arabica were analyzed, in order to assess their content of essential nutrients (macroand microelements) and the presence of potential bioactive compounds for the development of functional beverages. More in detail, elemental analysis using inductively coupled plasma atomic emission spectroscopy (ICP-AES) revealed a comparable elemental profile across both tissues and varieties. In parallel, liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) metabolomic analysis on the polar fraction was performed using both targeted and untargeted approaches, with methanolic and aqueous extractions to simulate the overall biochemical profile and the brewing process, respectively. Thus, a better understanding of the elemental profile and presence of phytochemical constituents could provide valuable insights into processing and extraction methods in order to preserve these compounds to better enhance the use of coffee leaves and berries in the formulation of new functional beverages. Acknowledgement: This work has been supported by METROFOOD-IT project which received funding from the European Union - NextGenerationEU, PNRR - Mission 4 “Education and Research” Component 2: from research to business, Investment 3.1: Fund for the realisation of an integrated system of research and innovation infrastructures – CLP IR0000033, CUP I83C22001040006 (D.M. Prot. n.120 del 21/06/2022). ADVANCES IN FOOD AND NUTRITION 49 Modulation of newborn and infant microbiota with prebiotics and probiotics Vera Neuzil Bunesova, Nikol Modrackova, Blanka Krausova, Eugenio Ingribelli, Jan Vinkler Czech University of Life Sciences Prague, Czech Republic Prebiotics and probiotics are now quite established terms that are associated with a beneficial effect on human health. But since when do they naturally become a part of our diet? Since when should we consider their supplementation? Moreover, it is necessary to consider not only their potential independent or synbiotic benefits, but also the interaction at the level of the host microbiota, which may be very individual. The aim of our work is to focus on the choice of suitable prebiotics and probiotics during the initial colonization of the newborn’s intestine, or rather the infant, with regard to gestational age, mode of delivery, and the initial period of nutrition, i.e., liquid food diet. Therefore, we longitudinally monitored an infant, combining both cultivation and molecular genetic approaches, to determine microbiota development and evaluate prebiotic and probiotic interventions. It is desirable to underscore the importance of targeted prebiotic and probiotic interventions during critical developmental periods, not only for their direct effects on specific microbial populations but also for their potential to modulate overall microbial interactions. Human milk is a unique source of oligosaccharides, which, however, despite the best efforts, cannot be fully synthesized, and therefore, we are looking for alternatives whose use has clear justifications. As well as including probiotics in the diet of newborns born out of right gestational age, surgically instead of vaginally, or whose microbial colonization is not standard and suffers e.g., from colic. These are just a few examples for considering the supplementation of prebiotics and probiotics. Moreover, the effect does not end with their application; it extends to the further development of the individual. Our findings highlight the importance of considering probiotic and prebiotic supplementation in newborns whose early microbial colonization may be suboptimal, such as those born via cesarean section, prematurely, or not breastfed. While breastfeeding remains the gold standard due to its unique oligosaccharide content, our results demonstrate that targeted microbiota modulation can have meaningful and lasting effects on infant development. Acknowledgements: This work was supported by a grant LUAUS23014 from the MEYS, Czech Republic and by the METROFOOD-CZ research infrastructure project (MEYS Grant No: LM2023064) including access to its facilities. ADVANCES IN FOOD AND NUTRITION 50 Microbial Spoilage Risks in Teran Wine: Tracking Brettanomyces and Biogenic Amines During Malolactic Fermentation Tatjana Radovanović Vukajlović1, Shanique Walker2, Julija Šupljika3, Mitja Martelanc4, Jan Reščič4, Melita Sternad Lemut4, Branka Mozetič Vodopivec4, Dejan Bavčar5, Klemen Lisjak5, Lorena Butinar4 1University of Nova Gorica, Wine Research Center, Slovenia 2DAFNAE, University of Padua, Italy 3Faculty of Food and Technology, University Josip Juraj Strossmayer Osijek, Croatia 4University of Nova Gorica, Slovenia 5Central Laboratories, Agricultural Institute of Slovenia, Slovenia Yeasts and lactic acid bacteria (LAB) are the main microorganisms involved in wine production, responsible for alcoholic fermentation and malolactic fermentation (MLF), respectively. During MLF, LAB convert L-malic acid into L-lactic acid, which primarily leads to biological deacidification but also results in changes in the wine’s aroma and microbiological stability. While LAB generally improves wine quality, certain strains can cause spoilage and are linked to the production of biogenic amines (BAs), which are nitrogenous compounds such as histamine, tyramine, and putrescine. Elevated levels of BAs may pose health risks, especially in sensitive individuals, potentially causing headaches, hypertension, and digestive issues. Similarly, the yeast Dekkera/Brettanomyces bruxellensis is considered a spoilage organism because of its ability to produce volatile phenolic compounds (e.g., 4-ethylguaiacol and 4-ethylphenol), which impart off-flavors resembling bandages, sweat, and smoke. As part of the AGROTUR+ project, we monitored LAB and B. bruxellensis populations, as well as BA and volatile phenol content, during MLF in Teran, a protected wine made from Refošk grapes in the Karst wine-producing region. The goal was to assess the presence of spoilage-related compounds and support improvements in the microbiological quality of this traditional wine. A total of 48 Teran wine samples from the 2023 and 2024 vintages were analyzed in this study. Volatile phenols were analyzed using HS-SPME-GC-MS, and BAs were determined via HPLC–UV following sample derivatization. The basic physicochemical parameters were determined using standard methods. LAB community and composition were analyzed through plating, 16S rRNA amplicon sequencing, and flow cytometry was used for LAB quantification in 2024 samples. In vintage 2024, B. bruxellensis was quantified in wine samples using plating and flow cytometry. Histamine was the most abundant BA (average 11.9 mg/L), followed by cadaverine (2.8 mg/L) and by putrescine (2.6 mg/L). There were differences between the vintages. No correlation was observed between total LAB counts and BA levels, indicating the need for further analysis of the LAB strain composition. For volatile phenols, 4-ethylphenol was primarily found at concentrations below its sensory threshold (600 µg/L), with only one sample exceeding this threshold. In contrast, 4-ethylguaiacol exceeded its threshold (110 µg/L) in four of the samples. B. bruxellensis was detected using the CyFlow™ BrettCount flow cytometry kit (Sysmex Partec), although no colonies were observed on the selective Dekkera/Brettanomyces differential medium (DBDM). Flow cytometry has proven to be a rapid and specific method for detecting spoilage yeasts, such as B. bruxellensis, with strong potential for early intervention and spoilage prevention in winemaking. ADVANCES IN FOOD AND NUTRITION 51 From concept to laboratory work: challenges and innovation in method validation for food additives Séverine Goscinny Sciensano, Belgium Validation of analytical methods is paramount to demonstrating their fitness for purpose. In the critical field of food additive analysis, this data underpins regulatory compliance, market surveillance, and public health risk assessments. The discipline is profoundly challenging, demanding that a single method be applicable across various food matrices, from simple beverages to complex food products. Concentrations of additives fluctuate dramatically, not only between different substances but also for the same additive across different food categories. Furthermore, the substances possess a broad and diverse range of physico-chemical properties (e.g. organic, inorganic, ionic, polar, and non-polar) requiring analysts to apply an exceptionally high level of technical skill to achieve methods with minimal variability and ensure data integrity. The rigorous validation process introduces further significant hurdles, including matrix coverage, broad concentration ranges, and substance stability. Failing to assess these factors properly threatens the reliability of the data. While validation is known to be costly and time-consuming, not doing it correctly risks wasting valuable time, money, and resources. To overcome these obstacles, we have developed a proactive and innovative approach. This strategic framework directly addresses food additive specificities, including the need for full matrix coverage and the use of additive blends (e.g. sweeteners, antioxidants and preservatives) which critically impact the required concentration range and limit of quantification. This novel methodology streamlines the validation process, significantly enhancing its efficiency without compromising the accuracy or reliability of routine data. Our presentation will detail this approach and its application to specific food additive classes, proving that innovation can thrive within the most stringent processes. 52 FOOD DATA ANALYSIS & FOOD PROCESSING Co-Creation, the responsibility to be innovative: the METROFOOD-IT case Giacomo Serafini1, Cesare Manetti2, Simone Cresti3, Valerio Miceli1, Gianluca De Rinaldis1, Claudia Zoani1 1ENEA - Italian National Agency for New Technologies, Energy and Sustainable Economic Development , Italy 2Sapienza University of Rome, Italy 3University of Siena, Italy The term “co-creation” refers to the collaborative efforts of various stakeholders—including businesses, universities, policymakers, and the general public—within innovation processes.[1]. In recent years, this approach has emerged as a response to a range of societal challenges, including environmental degradation, sustainability of industrial production, and issues related to food safety and food security. At the same time, co-creation is also increasingly recognised as an opportunity for making innovation processes more socially inclusive and responsible. Involving end-users, researchers, and stakeholders in the research process enhances project outcomes, aligning them more closely with societal needs and expectations, as emphasized by Horizon Europe’s First Pillar. This alignment boosts real-world applications and helps translate research findings into effective policy and practice. In fact, the so called “independent science”, when detached from its political and social context, often fails to provide valid solutions to tackle the complex problems faced by our society. Therefore, an engaging collaboration among researchers, policymakers, and citizens becomes essential because “by working together on the structuring of the problem definition after which a shared knowledge (shared reality’) can be achieved”[2,3]. The aim of this work is to explore the concept of co-creation within the European research landscape and to examine its transformative potential in shaping research and innovation. The goal is not only to highlight the benefits of co-creation, but also to propose a different perspective: to consider such a research approach as a responsibility rather than merely a strategic tool. To this end, the experiences developed within the METROFOOD-IT research infrastructure are presented, including co-creation strategies and initiatives undertaked so far. A focus is given on the role of Living Labs and Training & Education activities, which are crucial enablers for an inclusive and impactful research. Acknowledgment: This work was supported by METROFOOD-ITproject has received funding from the European Union - NextGenerationEU, PNRR - Mission 4 “Education and Research” Component 2: from research to business, Investment 3.1: Fund for the realisation of an integrated system of research and innovation infrastructures - IR0000033 (D.M. Prot. n.120 del 21/06/2022) References: [1] SCALINGS, Co-creating european future, Munich Center for Technology in Society Technical University of Munich, 2021 [2] Barbara J. Regeer and Joske F.G. Bunders, Knowledge co-creation: Interaction between science and society A transdisciplinary approach to complex societal issues, VU University Amsterdam Athena Institute, 2009 [3] R.J. In ‘t Veld, Willingly and Knowingly: The Roles of Knowledge about Nature and the Environment, Lamma Publishers, 2000 53 FOOD DATA ANALYSIS & FOOD PROCESSING AI-Driven Robotic Arm Guidance and Control for Agrifood: Enabling Smart, Zero-Touch and Safe Food Automation Lorenzo Coppola1, Rosario Schiano Lo Moriello2, Roberto Soldaini2, Francesco Bonavolontà2, Francesco De Santis1, Flora Amato2, Giorgio De Alteriis2, Mariateresa Caivano2 1A.C. S.p.A., Italy 2Universita degli studi di Napoli Federico ll, Italy Digital technologies, particularly those involving artificial intelligence, robotics and advanced sensing, offer concrete solutions to enabling smart, zero-touch and safe food automation, especially in sensitive production environments where hygiene and precision are paramount. In this context, the SCENARIO project (Advanced Robotic Solutions for the Agri-Food Industry) aims to design and validate intelligent technologies for advanced automation in the food and drink industry. The project core involves developing a smart robotic system for preparing baccalà (salt cod) batter. This system is designed to operate in controlled contamination environments and can consistently replicate a traditional process that is highly valued for its gastronomic qualities. Based on a multi-axis arm controlled by AI algorithms, the robot integrates computer vision and smart sensors to monitor critical parameters such as the position, attitude, appearance of the fish piece as well as viscosity of the batter in real time. The entire process — from mixing the ingredients to quality control — is automated thus lowing also to implement a zero-touch logic to minimize contamination risks and ensure the highest hygiene standards. Through a dedicated human-machine interface (HMI), operators can select recipes, program sequences and monitor system status. Environmental and process data collected by the sensors feed predictive modules that optimize operational efficiency by dynamically adjusting the robot’s behavior to real-world conditions. The SCENARIO project demonstrates how collaborative robotics can enhance tradition through technological innovation when applied to a typical food process such as baccalà batter production. The system is designed to be scalable and replicable for use in both small-scale local production and industrial lines. It is a concrete example of how digitalisation and automation can coexist with the cultural identity of quality food products. This work was supported by Campania Regional Program FESR 2021–2027, Campania Regional Program ESF+ 2021–2027, and POC 2014–2020, Specific Objectives 1.1 and 1.3, Actions 1.1.1 and 1.3.1 under Grant CUP: B15H23002630006 within the framework of the project SCENARIO, beneficiary company A.C. S.p.A. 54 FOOD DATA ANALYSIS & FOOD PROCESSING Toward Sustainable Mozzarella di Bufala Campana:LoRaWAN-Based Monitoring of Greenhouse Gases to Quantify and Reduce Emissions Michela Zampano, Francesco Cirillo, Francesco Bonavolontà, Maria Teresa Verde, Stefania Pindozzi, Ester Scotto di Perta, Alessandra Apostolico Universita degli studi di Napoli Federico ll, Italy The production of Mozzarella di Bufala Campana, a PDO-certified excellence of Southern Italy, is deeply rooted in centuries-old farming practices and a unique territorial heritage. Within this framework, ensuring animal welfare, environmental sustainability, and food safety has become essential. With increasing consumer demand for transparency and eco-responsibility, the implementation of technological solutions to ensure traceability and cutting emissions is essential [1]. This study introduces a wireless sensor network based on the LoRaWAN protocol [2], engineered for real-time monitoring of greenhouse gases—specifically carbon dioxide (CO₂) and ammonia (NH₃)—in buffalo barns. These gases are critical indicators of air quality, with direct implications for animal health, worker safety, and overall compliance. Long-term exposure to excessive concentrations can reduce productivity and compromise milk quality. Each sensor node integrates commercial electrochemical sensors (DOL-119 for CO₂ and DOL53 for NH₃), a low-power STM32 microcontroller, and an I-NUCLEO-LRWAN1 shield. The system operates autonomously, transmitting data at 10-second intervals to a central LoRaWAN gateway. Information is then processed via MQTT and visualized in Node-RED for real-time and historical tracking. The solution is designed for rural contexts, ensuring reliable transmission even in harsh barn environments lacking Wi-Fi or cellular infrastructure. The architecture was validated in a working buffalo dairy. Results confirmed stable communication and consistent data acquisition across all deployed nodes. Gas peaks were detected promptly, enabling timely ventilation. This improved environmental conditions and supported welfare and product quality. This work highlights how IoT technologies, integrated with Precision Livestock Farming practices, can modernize traditional dairy operations and help achieve European sustainability and food safety goals. Funding: This study was carried out within the Agritech National Research Center and received funding from the European Union Next-Generation EU (Piano Nazionale di Ripresa e Resilienza (PNRR) – Missione 4, Componente 2, Investimento 1.4 – D.D. 1032 17/06/2022, CN00000022). Acknowledgment: METROFOOD-IT (Next Generation EU, PNRRM4C2, Investment 3.1: Fund for the realization of an integrated system of research and innovation infrastructures-IR0000033D.M. Prot. n.120 del 21/06/2022) This work was supported by METROFOOD-IT which received funding from the NRRP— Mission 4 ‘Education and Research’, Component 2: From Research to Business, Investment 3.1: Fund for the creation of an integrated system of research and innovation infrastructures—IR0000033 (MD No. 120 of June 21, 2022). [1] Tullo, E., Finzi, A., and Guarino, M., Science Total Environ. 650, 2751 (2019) [2] Haxhibeqiri, J., De Poorter, E., Moerman, I., et al., Sensors 18, 3995 (2018) 55 FOOD DATA ANALYSIS & FOOD PROCESSING Explainable Machine Learning on SWIR Hyperspectral Data for Origin-Based Classification of Wheat Grains and Flours Michele Magarelli1, Florinda Artuso2, Pierpaolo di Bitonto1, Alessandra Pasquo2, Donato Romano3, Pierfrancesco Novielli3, Antonia Lai2, Sabina Tangaro1 1Università degli Studi di Bari Aldo Moro, Italy 2ENEA, Centro Ricerche Frascati, Italy 3INFN, Sezione di Bari, Italy The authentication and traceability of agro-food products are increasingly critical for ensuring food quality, combating fraud, and promoting transparency in supply chains. In the case of durum wheat—a staple crop with strong geographical and varietal identity—traditional analytical methods are often limited in throughput, scalability, and interpretability. In this study, we propose a novel approach that integrates short-wave infrared (SWIR) hyperspectral imaging (900–1700 nm) with machine learning (ML) and explainable artificial intelligence (XAI) to classify durum wheat samples by geographical origin and variety. A total of 16 wheat samples were collected from multiple Italian regions and cultivars. Each sample was analyzed in both whole-grain and flour form using the NIREOS HERA SWIR-2 hyperspectral system. The resulting high-dimensional data were normalized and preprocessed to extract meaningful spectral features. We then applied supervised ML algorithms—including Support Vector Machines (SVM), Random Forest (RF), and Partial Least Squares Discriminant Analysis (PLS-DA)—to classify the samples. To move beyond black-box predictions, we incorporated explainable AI techniques, primarily SHapley Additive exPlanations (SHAP), to uncover the most influential wavelengths and spectral intervals contributing to the model’s decisions. These insights not only confirmed known chemical-spectral associations but also revealed novel discriminative features relevant to wheat origin. The proposed pipeline achieved high classification accuracy and provided interpretable models that support the development of AI-based tools for rapid, non-destructive screening in cereal quality control. This study highlights the potential of combining hyperspectral imaging with interpretable machine learning as a robust solution for real-time food authentication, aligning with the goals of infrastructures like METROFOOD-IT and the broader vision of open, data-driven agro-food systems. 56 FOOD DATA ANALYSIS & FOOD PROCESSING Comparative analysis of volatile compounds and sensory assessment of olive oils Erika Bešter, Albert Kravos, Gašper Kozlovič Science and Research Centre Koper, Slovenia The accurate detection of volatile compounds (VOCs) and sensory defects in virgin olive oil is very important for characterisation of virgin olive oil as well as to support olive oil classification according to Commission Delegated Regulation (EU) 2022/2104. This study is aimed to validate methods for determination of VOCs and to determine which of the two used methods is more reliable for individual marker. A set of 18 VOCs was set based on previous studies which demonstrated that these 18 compounds showed the strongest correlation with sensory defects. In this study, 20 virgin olive oil samples were determined by SPME-GC-MS and by SPME-GC-FID and compared with sensory analyses. Despite progress in analytical instrumentation, inter-laboratory variability and mismatches with sensory evaluation persist. VOC data, z-scores, and classification values from all panels were assessed. Bias, standard deviation, and failure rates were explored, and the most divergent compounds were compared against established sensory defect markers. The performance of our sensory panel in the interlaboratory comparison was excellent. Comparison of VOCs assessment results showed that our laboratory’s results for certain parameters, such as ethyl acetate, were in close agreement with consensus values. On the other hand, some other parameters, such as aldehydes and acids, that correlate with rancid and musty defects, were underestimated. It turned out that for some markers, SPME-GC-MS method provided better results, while for other markers, the SPME-GS-FID method was more suitable. Ultimately, our findings support the implementation of enhanced methods for sensory defect-related VOCs. 57 FOOD DATA ANALYSIS & FOOD PROCESSING Comprehensive analysis of volatile compounds released during the heattreatment of highly processed plant-based alternatives Elnora Abdullajeva1, Tania Portolés Nicolau2, Juan Vincente Sancho2, Elena Hakme1, Lene DuedahlOlesen1 1Technical University of Denmark, Denmark 2Jaume I University, Spain Plant-based alternatives is a relatively new group of products with a diverse composition and varying degrees of processing. While the presence of these products on the market has been steadily growing, there is still limited information on the composition of chemical profiles of these products, especially the ones generated through heat-treatment. In this study, we aim to describe the chemical profiles of volatile organic compounds released during heat-treatment of plant-based meat and cheese alternatives. The extraction strategy was based on three distinct approaches: (1) solvent extraction (SE) using acetonitrile followed by a QuEChERs-based clean-up, (2) headspace-solid phase microextraction (HS-SPME) using a CAR/PDMS/DVB fiber, and (3) dynamic headspace with sorbent trapping followed by thermal desorption (DHS-TD), which is less commonly applied in food studies. For the latter method, two DHS-TD tube types were compared: TenaxTA and a two-phase Bio-monitoring sorbent tube (non-disclosed composition, produced by Markes International). Additionally, two different volatile phases were sampled: i) the one released during the processing (150°C, 20 min) and ii) a post-processing phase (50°C, 30 min). In all cases, the analysis was performed on GC-QOrbitrap hyphenated with TD100-xr system for thermal desorption, followed by the processing with a non-target workflow (Compound Discoverer, version 3.3.3). Volatile phases collected using sorbent tubes (during and post-processing) were compared with the chemical composition found using SE and SPME [1]. Regarding DHS-TD, TenaxTA and Bio-monitoring sorbents yielded similar profiles, though better reproducibility was observed using TenaxTA (94 versus 79 features with RSD<20%). The principal component analysis (PCA) of TenaxTA data showed clear separation between duringand post-processing phases, with greater variability in triplicates during processing. Volatile profiles released during processing were mostly comprised of O-containing aromatic and cyclic compounds, followed by N-containing compounds post-processing. As for the other techniques, SE revealed more features overall, but SPME highlighted the heterogenicity of volatiles released by different plant-based and cheese alternative matrices. Both techniques showed good reproducibility and will be used in the future as complementary approaches to DHS-TD extraction using TenaxTA sorbent tubes for analysis of commercially available products. References [1] S. Mokh, L. Lacalle-Bergeron, D. Izquierdo-Sandoval, M. C. Corell, J. Beltran, J. V. Sancho, and T. Portolés, Food Chem. 449, 139312 (2024). 64 New trend in Multiresidue Methods: Achieving the maximum with the minimum Amadeo R. Fernández-Alba European Reference Laboratory for Pesticide Residues in Fruits and Vegetables University of Almería, Spain The evolution of food analytical chemistry is continuous, delivering innovative solutions to classical challenges—low limits of quantification (LOQ), broad analyte scope, automation, and organic waste minimization. In routine laboratory management, accurately assessing your needs is essential for tailoring the adoption of these technologies, which may be constrained by time, accreditation requirements, and available resources. The primary objective remains meeting stringent food safety and trade standards, while also embracing ever more demanding measures that safeguard both human health and the environment. In recent years, key advances include: - Quantification of pesticides and related food contaminants down to low ppb levels - Screening hundreds of compounds with just a few multi residue and single residue methods - Turnaround times of 24 hours or less However, concerns over food security and the demands of expanding international trade are driving the need for even stricter contaminant controls measures that not only uphold the highest standards for human health but also address environmental impacts. During this talk, we’ll showcase several recent technological developments in chromatography and mass spectrometry, alongside emerging requirements such as reducing organic waste and the increasing importance of monitoring PFAS contaminants. 65 Stable isotope dilution assays for mycotoxins help assess the risk of plant-based meat alternatives Michael Rychlik Technical University of Munich - TUM, Germany Worldwide dietary changes are partly due to consumers questioning the traditional production and consumption of animal-based food for its impact on animal welfare, energy waste, loss of biodiversity, and last but not least being detrimental to human health. Therefore, alternatives to traditional animalbased foods are increasingly developed, marketed and consumed. The range of products covers plantbased meat and milk alternatives along with products elaborated from cellular agriculture or tissues produced by protein-based additional manufacturing. We have been investigating plant-based meat alternatives for their mycotoxin content using table isotope dilution assays and have performed a risk evaluation. The outcome is giving a different perspective on the narrative that all of these alternatives are beneficial for ˝one health˝. 66 Building trust in the whitefish value chain: Organisational and technical dimensions of a traceability pilot Kine Charlotte Jakobsen, Truls Bakkejord Ræder SINTEF Nord, Norway The whitefish value chain faces persistent challenges related to fragmented data flows, limited transparency, and vulnerability to food fraud. These issues undermine regulatory oversight, hinder sustainable management of fish stock, and erode consumer trust. While emerging technologies like AI, DLT, and IoT can improve traceability, their impact and effectiveness depend not only on technical performance, but also coordination of actors, shared standards, and data governance. The pilot “Combating white fish counterfeiting in Norway”, part of the Horizon Europe project WATSON (grant id: 10071171), currently under implementation in Norway, Denmark, and Poland, explores how a digitally enhanced traceability system can foster both consumer and regulatory trust in the seafood sector. The system integrates sensor data, a DLT-based platform, and Digital Food Product Passports (DFPP) based on GS1/EPCIS standards, enabling end-to-end data visibility, from catch to consumer. In parallel, an Early Warning System (EWS) is being developed in cooperation with control authorities to detect anomalies and potential fraud risks. Beyond its technical demonstrators alone, the pilot highlights the importance for joint governance in data-sharing infrastructures, including clearly defined roles, accountability of data input and quality, and mutually trusted rules for access and use. A key use case involves the development of a premium, single-supplier product line between the fishing vessel Hermes, and the processor Espersen. This coordinated supply chain enables full-cycle traceability testing, controlled and iterative feedback loops, and enhanced legitimacy among consumers and regulators. Consumers access product data through DFPPs, while regulatory bodies test the EWS for real-time oversight. Initial findings from stakeholder workshop and implementation planning suggest that trust is shaped by three interrelated factors: Data reliability (technical), process transparency (organisational), and role clarity (governance). Regulatory stakeholders value more continuous and granular access to supply chain data, while industry actors emphasise interoperability, proportionate data sharing, and protection of commercial confidentiality. Rather than treating technology as a standalone solution, the project underscores the significance of organisational and collaborative structures in making digital traceability credible and scalable. While core technologies are increasingly mature, major barriers lie in embedding them within real-world settings across diverse organisational contexts. Accordingly, digital traceability is framed not merely as technological innovation, but as a socio-technical transformation that requires institutional adaption, trust-building, and sustained coordination. Its success depends as much on governance and aligned incentives as on data standards and interoperability. CLUSTERING 67 CLUSTERING Consumer Purchase Intentions for BlockchainTracked Olive Oil and Feta Cheese Seyyedehsara Sadrmousavigargari, Giulia Maesano, Alessandra Castellini Alma Mater Studiorum – Università di Bologna, Italy Introduction: Blockchain technology appeared as a novel and promising way to address product quality, provenance, and food safety issues. Its main feature is the creation of a decentralized and immutable record that enables the secure and transparent exchange of data between the parties involved [1]. Previous studies have investigated the capabilities of blockchain technology (BCT) [1, 2]. However, research on consumer attitudes and purchase intentions for products tracked using blockchain technology has been limited. To better understand consumers’ intentions to purchase products tracked through a blockchain system, this research assesses consumers’ intentions to buy olive oil in Italy and feta cheese in Greece. Data and Framework: Data collection was carried out via a consumer panel of a market research institute with a quota sampling method for age and gender groups. A total of 1000 responses were collected from Italian and Grecian adults. The questionnaire, grounded in TPB constructs, also incorporated items addressing consumer trust in quality certifications and their attitudes toward technology. Results: Structural equation modeling (SEM) was employed to examine the factors influencing consumers’ intentions. The PLS-SEM analysis showed that subjective norms, perceived behavoural control, attitudes toward the behaviour of buying blockchain-traced products and Attitude toward technology significantly impacted consumers’ intentions to purchase blockchain traced olive oil and feta cheese. However, trust in quality certifications did not significantly influence purchase intentions. Conclusion: The findings, based on two typical products in two different European countries, suggest that successful marketing strategies should focus on informing consumers about the benefits of blockchains, simplification of the user experience and use of social influences to promote the adoption of blockchain traceability. References: Reitano, M., Pappalardo, G., Selvaggi, R., Zarbà, C., & Chinnici, G. (2024). Factors influencing consumer perceptions of food tracked with blockchain technology. A systematic literature review. In Applied Food Research (Vol. 4, Issue 2). Elsevier B.V. Kamilaris, A., Fonts, A., & Prenafeta-Boldύ, F. X. (2019). The rise of blockchain technology in agriculture and food supply chains. Trends in food science & technology, 91, 640-652. Galvez, J. F., Mejuto, J. C., & Simal-Gandara, J. (2018). Future challenges on the use of blockchain for food traceability analysis. TrAC Trends in Analytical Chemistry, 107, 222-232. 68 CLUSTERING Blockchain based supply chain for PDO Feta Cheese: The ALLIANCE approach Kostas Choumas, Apostolis Apostolaras, Stavroula Maglavera, Thanasis Korakis University of Thessaly, Greece ALLIANCE is a European Horizon Europe initiative designed to enhance seven Food Supply Chains (FSCs), including one that facilitates the manufacturing and delivery of PDO Feta Cheese. ALLIANCE has developed an architecture that offers a thorough implementation plan, including the complete range from data acquisition to data utilization. IoT networks provide data collection via a modular architecture, permitting the dynamic integration of diverse sensor devices. The collected data are processed and stored before being utilized by the apps that support FSC administration. Blockchain is the essential technology utilized for safe and immutable data storage in a commercially viable manner. All stakeholders involved in the FSC contribute to the Blockchain network and are certain that no person can derive illicit advantages from data manipulation. The problem is that data storage in a Blockchain network presents performance issues. Efficient data storage requires the effective management of data, distinguishing the information intended for Blockchain from those stored in an auxiliary database referred to as off-chain. The simultaneous data storage in both Blockchain and off-chain systems is intricate, as maintaining consistency between the two repositories is crucial, requiring the deletion of data from one database if the other fails. In ALLIANCE, we engaged in iterative communication with stakeholders linked to each FSC to formulate its respective data model and ascertain the critical data to be preserved on the Blockchain, while the rest is retained off-chain. Additionally, we utilized the GS1 EPCIS protocol to standardize the storage and presentation of data of each FSC, hence enhancing interoperability among the FSCs. Finally, the UI of the ALLIANCE platform is a web-app that gives users the ability to interact with the data produced by the system. The administrators of the FSCs are provided with a dashboard that allows them to monitor their current state. This includes monitoring the materials that are present at each stage, the way they are processed, the outcomes of the relative quality controls, and the overall conditions of the FSCs. In addition, it serves as the point of input for the data that is generated by the users of the system, which includes farmers, drivers, people responsible for storage, and other individuals. 69 CLUSTERING Securing Agri-Food Supply Chains with AI for Early Warning, Fraud Detection, and Decision Support Pantelis Lappas1, Amalia Ntemou2, Spyros Evangelatos2, Filodamos Papanatsios2 1University of the Aegean and Research and Innovation Development Department, Netcompany S.A., Luxembourg 2Research and Innovation Development Department, Netcompany S.A., Luxembourg The AI-enabled Early Warning and Decision Support System (EWDSS), developed within the ALLIANCE project, has proven to be a powerful system for detecting and managing food fraud across diverse agrifood supply chains. The system integrates predictive analytics, fuzzy logic, and multi-criteria decision analysis (MCDA) techniques to support proactive risk management and informed stakeholder decisionmaking. The early warning component leverages fuzzy logic to synthesize multiple risk indicators into a comprehensive fraud score. In the Organic Pasta supply chain, the system evaluates contamination levels across production stages (wheat, semolina, and pasta), field audit compliance rates, and certification validity, each mapped to fuzzy sets and aggregated into an overall fraud label. In the Feta Cheese supply chain, the system monitors key PDO-related parameters, specifically pH and temperature values, to identify potential adulteration or quality deviations early in the process. To support fraud prediction, a synthetic dataset simulating four years of Feta Cheese supply chain operations was developed. It captures complexities such as milk collection, transport, testing, supplier risk profiles, and realistic fraud injection mechanisms (e.g., cow milk adulteration, water dilution, and excessive goat milk in sheep milk). A key feature of the EWDSS is a multi-criteria evaluation framework for selecting optimal machine learning models using multiple performance metrics (e.g., accuracy, precision, recall, etc.). Based on the TOPSIS method, this framework objectively ranks supervised classifiers and unsupervised anomaly detection models applied in the Fava Beans and Organic Honey supply chains, using spectral, DNA, and sensor data. Complementing automated analytics, a human-in-the-loop decision support tool enables stakeholders to structure complex problems hierarchically, express preferences, assess consistency, and transparently rank alternatives. Use cases from the Fava Bean supply chain illustrate prioritization of fraud types by impact and detection difficulty, as well as evaluation of mitigation strategies based on practical and economic considerations. Implemented using modern technologies such as Python, FastAPI for RESTful services, Apache NiFi for data integration, and Apache Superset for dashboarding, the EWDSS delivers a robust AI-driven solution for early warning, predictive modeling, model evaluation, and participatory decision support in food fraud risk management. 70 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Exploitation of seaweed for food and feed applications – the need for elemental analysis for assessment of safety Jens SLOTH, Susan Holdt National Food Institute, Technical University of Denmark, Denmark Seaweed is the common term for marine macroalgae plants, which may be divided into green, red and brown algae types. There is a growing interest in increasing the exploitation of marine macroalgae for commercial purposes including their uses in relation to food and feed production and thereby contribute to the green transition by exploitation of this under-utilized bioresource. Many seaweeds have the potential to accumulate various trace elements and contain consequently relatively high levels of both essential and potentially toxic elements (PTEs). There is consequently a need to document the levels of potential toxic elements in seaweeds and to ensure that the use/consumption of the seaweeds is safe. In this context, a better understanding of how biological and environmental factors, like seaweed type, location, sampling season etc. affects the levels of trace elements is called upon, in order to be able to select seaweeds with optimum characteristics for commercial use. Additionally, further knowledge and understanding of the effect of post-harvest processing on the levels of elements in the biomass is called upon. Currently, the European Commission is considering setting future maximum levels for certain PTEs (incl. Cd, Pb, Ni, I, iAs) in seaweed and consequently there is a need to ensure the analytical capability of control laboratories for the future compliance testing. This will e.g. include harmonization of methods, development of reference materials, conduction of proficiency testing schemes (PTs). The lecture will present the outcome of recent research projects related to seaweed use in food and feed applications conducted at the National Food Institute at the Technical University of Denmark (DTU Food) with a focus on the analytical procedures used for the determination of trace element (including trace element species) and the subsequent evaluation of the obtained data in relation to the assessment of food and feed quality and safety. The lecture will further address the current status on element determination in seaweeds from the point of view of the EU reference laboratory for metals and nitrogenous compounds in feed and food (EURL-MN) hosted by DTU Food. 71 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Development of an UHPLC-MS/MS method for determination of ultra-short PFAS in vegetable samples Michela Torrelli1, Consolato Schiavone1, Francesco Romaniello1, Alberto Salomone2, Andrea Mario Rossi1, Chiara Portesi1 1Istituto Nazionale di Ricerca Metrologica (INRiM), Italy 2University of Torino , Italy PFAS, a large group of widely used synthetic compounds, are known for high resistance to water, fat, and heat. Defined as environmentally stable “forever chemicals,” their bioaccumulation and potential toxicity pose growing human health concerns. Tomatoes can accumulate high PFAS both from contaminated soil and irrigation water, becoming a primary food chain entry. In particular, matrix high-water content could facilitate the presence of highly polar ultra-short-chain (USC) PFAS. Monitoring health risks associated with PFAS exposure requires accurate and harmonized analytical methods. With common reverse-phase chromatography methods, poor or no retention of USC-PFAS causes poor reproducibility of retention time and, consequently, inaccurate quantification. This work aims to develop a ReversedPhase Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry method capable of efficient retention and separation of USC-PFAS, as well as addressing the problems associated with these contaminants in laboratory environments. The investigation focused on DFA, TFA, PFPrA, PFBA, PFMeS, PFEtS, PFPrS, and PFBS. Analysis was performed with UHPLCTRIPLE QUAD (SCIEX 6500+) on a variety of tomato products (concentrate, puree, and pulp) to quantify the possible presence of PFAS in the matrix. The optimization of the chromatographic conditions and the use of a delay column allowed efficient chromatographic separation. High sensitivity and good linearity were demonstrated in both solvent and matrix curves, with R2 ≥ 0.99 obtained for both considered concentration ranges (0.100 µg/L to 10 µg/L for DFA, TFA, and PFPrA; 0.010 µg/L to 1 µg/L for PFBA, PFMeS, PFEtS, PFPrS, and PFBS), corresponding to instrumental LOQs of 0.100 µg/L and 0.010 µg/L, respectively. The RSD% values, below 5% over 35 injections of both in solvent and in matrix standard mixtures, confirm the great repeatability and reproducibility of the developed method. The final evaluation confirmed extraction efficiency and quantification for USC-PFAS in tomato samples, showing recoveries and RSD% compliant with EU guidelines. Only PFBA was detected in screened samples—concentrate (0.265 ± 0.038 µg/kg), pulp (0.056 ± 0.005 µg/kg), and puree (0.063 ± 0.001 µg/kg)—suggesting its endogenous content increased during industrial processing. Acknowledgments: The present work was performed in the IMPreSA (Metrological Infrastructure for Food Safety) infrastructure and has been supported by the project “Analytical Contaminants Assessment in Food”, which has received funding from the Fondazione Compagnia di San Paolo, and the project 23IND13 ScreenFood, which has received funding from the European Partnership on Metrology, cofinanced by the European Union’s Horizon Europe Research and Innovation Programme and by the Participating States. 72 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Chemopreventive Effects of Rosmarinic Acid against the Carcinogenic Food Contaminant Aflatoxin B1 Matjaž Novak1, Veronika Furlan2, Martina Štampar1, Alja Štern1, Bojana Žegura1, Urban Bren2 1National Institute of Biology, Slovenia 2Faculty of Chemistry and Chemical Engineering, University of Maribor, Slovenia Rosemary (Rosmarinus officinalis L.) has long been valued for its therapeutic properties. This highly valued medicinal plant contains various natural bioactive compounds that exert diverse pharmacological effects. One of the key bioactive compounds is rosmarinic acid (RA). Available studies suggest that RA could be considered a potential anti-cancer agent in various cancer treatments, and its chemoprotective properties also highlight its potential in safeguarding against foodborne carcinogens. Therefore, in the present study, the chemopreventive effects of RA against the carcinogenic food contaminant aflatoxin B1 (AFB1) were investigated using both in silico and in vitro approaches. The in silico investigation of the chemical reactions between rosmarinic acid and the ultimate chemical carcinogen of AFB1, aflatoxin B1 exo-8,9-epoxide (AFBO), was conducted by calculating activation free energies with advanced DFT methods M11-L and MN12-L, in combination with the flexible 6-311++G(d,p) basis set and implicit Solvation Model Density (SMD), according to the newly developed QM-CSA protocol. Following computational analyses, the chemoprotective effects of RA were further studied in vitro in human hepatocellular carcinoma (HepG2) cells by analyzing its influence on AFB1-induced genotoxicity using the comet assay, γH2AX, and pH3, while its impact on cell proliferation and cell cycle modulation was assessed using flow cytometry. Our computational results revealed that the activation free energy required for the reaction of RA with AFBO is significantly lower than the activation free energy for the competing reaction of AFBO with guanine, which indicates that RA acts as an efficient scavenger of AFBO, potentially preventing AFB1-specific DNA adduct formation. The chemoprotective activity of RA was confirmed through in vitro experiments, which demonstrated a reduction in AFB1-induced DNA singleand double-strand breaks in HepG2 cells exposed to a mixture of AFB1 and RA at non-cytotoxic concentrations. In addition, RA reversed the AFB1-induced reduction in cell proliferation. Given the widespread occurrence of AFB1 in contaminated food, the chemoprotective effects of RA could have significant implications for mitigating the health risks associated with chronic AFB1 exposure and represent a potential dietary intervention strategy to reduce cancer risk. Further studies are needed to uncover the precise mechanisms underlying its effects, as well as in vivo studies to confirm its anti-cancer effects and to better validate its role in preventing AFB1-induced carcinogenesis. Acknowledgements: Z4-60176, L7-60161, J160001, P1-0403, L2-4430, J2-60044, J160001, J1-60016. 73 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS METROFOOD-BE service provision to support circular economy, food safety and sustainable solutions for food packaging Joris Van Loco Sciensano, Belgium Aim: This work presents two research use cases under METROFOOD-BE to showcase how Belgian infrastructure supports metrology-driven food safety research. The first use case focuses on compiling a comprehensive database of potentially migrating chemicals from paper-and-board food contact materials and evaluating their hazard and risk. The second use case characterises zinc oxide (ZnO) nanoparticles of a novel bio-based packaging system. By demonstrating database development, risk assessment and ongoing analytical workflows, we highlight how METROFOOD-BE enables reliable measurement science for food packaging safety. Method: In use case 1, a substance database has been built to include 8 188 unique entries compiled from authoritative sources, such as the ESCO working group on paper and board, the Swiss ordinance on food contact materials, the council of Europe synoptic document (2005), the council of Europe resolution on paper and board (2024), EU regulation No 10/2011 on plastics, and relevant scientific literature. Each entry contains identifiers (CAS number, InChIKey), and when available, the reported use or occurrence, and available toxicological annotations. This list was filtered down through regulatory, exposure, and toxicological considerations to contain a more manageable database of 272 substances for detailed hazard classification via the SILIFOOD tool and further preliminary risk assessment. In use case 2, biodegradable films were formulated by embedding ZnO nanoparticles, olive leaf extract, and pine resin into a microfibrillated cellulose matrix. Physicochemical characterisation based on ultrathin sectioning of the films followed by scanning transmission electron microscopy (STEM) aims to visualise the distribution of nanoparticles within the film and to assess their size distribution. Parallel ICP-OES analyses are performed to quantify total ZnO vs nano partical ZnO in migration simulants under standard testing conditions. Results: Both use cases were selected through an open call based on technical merit, innovation potential, and infrastructure availability. To date, the substance selection for use case 1 is fully compiled and underwent SILIFOOD-based hazard classification. The substance selection will now undergo risk assessment through exposure scenarios. For use case 2, sample preparation has been completed, STEM imaging is ongoing to assess ZnO nanoparticle size distribution, and migration experiments with the related ICP-OES measurements are in the final phase. Conclusion: The creation of a large-scale, curated substance database and the initiation of physicochemical analyses for a biobased nanocomposite demonstrate how METROFOOD-BE’s services underpin reproducible measurement workflows. These ongoing efforts will inform subsequent prioritisation and safety evaluation, ultimately guiding risk assessment protocols and fostering safer, more sustainable food packaging solutions. By providing open access to these metrological capabilities, Belgium reinforces its leadership in European food safety innovation. 80 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Assessment of the impact of cooking on the fate of potentially toxic elements and nutrients in foodstuff Thinhinane HAMITOUCHE1, Jens SLOTH2, Axelle LEUFROY1, Petru Jitaru1 1French Agency for Food, Environmental and Occupational Health and Safety, France 2National Food Institute, Technical University of Denmark, Denmark Current approaches typically used in chemical risk assessment related to food consumption rely almost exclusively on the information retrieved based on the composition of the raw food items. However, various cooking procedures may influence the chemistry of the contaminants, including potentially toxic trace elements (PTTEs), as well as the nutritional quality of food. Actually, the assessment of the impact of food cooking on the fate of both toxic and essential inorganic chemical species is scarcely carried out and knowledge in this field is lacking. The present study aims to contribute to a better understanding of the impact of the on the fate of potentially toxic trace elements such as Pb, Cd, Hg, As, Cr, Al, Sb, Mn, Co and Ni and the speciation analysis of As and Hg (in both cases, inorganic and organic species will be studied) in food. For this purpose, various food matrices of vegetable origin (lentils, rice) and animal origin (beef steak, beef liver, salmon and swordfish) were subjected to different domestic cooking methods depending on their type (these included e.g. boiling, steaming, deep-frying, and oven baking). Then, raw and cooked samples were analysed in parallel to evaluate the impact of the cooking procedure on the concentrations of different TE (potential losses, transformations, or enrichments). In order to accurately assess the impact of food cooking on the risk/benefit balance, the variation in levels of the most important nutriments, such as P, S, I, Na, K, Ca, Mg, Se, Fe, Mo, Cu and Zn were also investigated. The analytical approach was based on the quantification of total levels of 22 elements by ICP-MS and speciation analysis of As and Hg by HPLC-ICP-MS. Preliminary results in terms of lentils and rice cooking impact showed limited changes in total concentrations for most elements. A slight increase in chromium was observed in lentils, while the levels of other elements remained stable. In rice, reductions were noted for several elements, with inorganic arsenic decreasing, consistent with existing literature. 81 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Oxidative Stress and Inflammation Dynamics in Type 2 Diabetes: Effects of the SGLT2 Inhibitor Empagliflozin Giorgi Goderdzishvili Czech University of Life Sciences Prague, Czech Republic This study examines oxidative stress and chronic inflammation, key factors in type 2 diabetes development. Oxidative and nitrative stress results from an imbalance between reactive oxygen species (ROS) and antioxidants, causing cellular damage. Chronic low-grade inflammation involves elevated proinflammatory cytokines that impair insulin signaling and metabolism. Hereditary Hypertriglyceridemic (HHTg) rats, a non-obese model prone to hypertriglyceridemia, insulin resistance, low HDL, fatty liver, and Wistar rats were fed a standard chow diet and treated with empagliflozin. Urine samples from four groups were analyzed: HHTg-Control, HHTg-EMPA, Wistar-Control, and Wistar-EMPA. Significance was tested within each strain (Control vs. EMPA) and between strains under EMPA treatment (Wistar vs. HHTg). Results showed significant differences in oxidative stress biomarkers between EMPA-treated and untreated groups. Lipid peroxidation markers changed notably: 8-isoprostane levels differed between Wistar-EMPA vs. HHTg-EMPA (P < 0.0001) and HHTg-Control vs. HHTg-EMPA (P < 0.05). MDA levels were also higher in treated rats (P < 0.01 and P < 0.05). Nucleic acid damage markers were elevated in HHTgEMPA vs. Wistar-EMPA: 8-OHdG (P < 0.01), 8-OHG (P < 0.0001), and 5-HMU (P < 0.01). Peptide and nitrative damage biomarkers increased in HHTg-EMPA vs. HHTg-Control: o-tyrosine (P < 0.01), and between HHTg-EMPA vs. Wistar-EMPA for 3-nitrotyrosine (P < 0.001) and 3-chlorotyrosine (P < 0.01). Inflammatory biomarkers significantly decreased after empagliflozin treatment. In Wistar (Control vs. EMPA), leukotriene C4, IL-8, IL-6, and leukotriene B4 all declined (P < 0.05), leukotriene E4 and Leukotriene D4 (P < 0.001). In HHTg (Control vs. EMPA), leukotriene C4 and IL-8 decreased (P < 0.05), IL-6 declined (P < 0.01), while leukotriene D4 and E4 showed no change (P = 0.94 and P = 0.357). Empagliflozin reduces inflammatory biomarkers despite increased oxidative stress, which may limit pro-inflammatory cytokine activity by modifying their precursors and be influenced by deficiencies in daily dietary variability. Antioxidantrich functional foods could manage oxidative stress and inflammation in type 2 diabetes, preventing further comorbidities. Supported by OMICs techniques like UHPLC-QqQ-MS and GC-MS and advanced statistical analyses, this approach proves valuable for early prognosis of complex pathologies. The strong association between metabolic syndrome (MetS) and chronic kidney disease (CKD) may be explained by shared mechanisms including chronic inflammation, oxidative stress, and insulin resistance. 82 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Quality of tomatoes grown in sludge containing contaminants of emerging concern Anja Vehar1, Nina Kacjan Maršič2, Lidija Strojnik1, David Heath1, Ester Heath1, Nives Ogrinc1 1Jožef Stefan Institute, Department of Environmental Sciences, Slovenia 2University of Ljubljana, Biotechnical Faculty, Department of Agronomy, Slovenia Aerobic biological wastewater treatment generates substantial quantities of sludge as a by-product, which poses challenges for its disposal and management due to its volume. Moreover, while sludge contains nutrients and organic matter, it frequently contains pathogens, toxic elements, hazardous chemicals, contaminants of emerging concern (CECs) and microplastics. If not properly treated or disposed of, these constituents pose serious environmental and public health risks, including the potential contamination of soil and water resources [1]. Anaerobic digestion is a common method for processing sludge. The technique employs anaerobic microbes under oxygen-free conditions to stabilize sludge, reduce its volume, and generate biogas (methane), which can serve as a source of energy [2]. However, the final digestate still requires dewatering and environmentally safe disposal. Identifying sustainable and safe methods for sludge disposal remains a challenge, as current practices, such as incineration, landfilling, and land application, are each associated with specific limitations, including environmental pollution, land use constraints, disposal costs and regulatory restrictions. [3]. In this study, the use of anaerobically digested sludge as fertiliser in tomato (Solanum lycopersicum L., cv. Rally) cultivation was investigated through pot experiments assessing metabolomic changes in tomato fruits due to exposure to sludge. Tomatoes were grown in various substrates: peat; peat spiked with CECs at two concentrations (27 CECs, 0.16 and 1.7 mg/kg); peat amended with dried sludge at two application rates based on N and K requirements; peat spiked with dried sludge (27 CECs, 11 mg/kg, sludge added according to N and K requirements) and peat mixed with non-dried sludge (1:1). Tomato fruits were analysed for quality parameters, including sugars (HPLC-RI), organic acids (HPLC-UV), polyphenols (HPLCDAD), amino acids (GC-MS), fatty acids (GC-FID), volatile organic compounds (HS-SPME GC-MS) and elemental composition (ICP-MS). Preliminary results revealed significant differences in sugars, organic acids, carotenoids, polyphenols, and volatile organic compounds. Significant metabolic changes indicate that exposure to sludge and CECs triggers stress responses in tomato fruits. Acknowledgements: This study was performed with the financial assistance of the Slovenian Research Agency and Innovation Agency – ARIS, namely Program Group P1–0143, P4–0013 and P4–0085; projects L7–4422 and J2–4427, and the Infrastructural Centre IC RRC-AG (IO-0022–0481-001). Anja Vehar acknowledges ARIS for the Young Researcher grant. References: 1. Li, Z., Yuan, D., Ecotoxicol. and Environ. Saf., 281, 116597 (2024) 2. Ahlawat, S., Nandal, M., Springer Singapore, pp. 369–379 (2020) 3. Rizzardini, C., Goi, D., Sustainability, 6, 2424–2434 (2014) 83 ANALYTICAL APPROACHES IN FOOD CONTAMINANTS Natural actives for ecofriendly/sustainable alternative disinfection strategies for agricultural products Rodolfo Esposito University of Siena, Italy The society and the habits of people are progressively changing as well as the request on global market. Indeed, the increasing frenzy of modern lifestyle prompted the consumers looking for fresh, healthy, and ready-to-eat foods. Nowadays a high attention is given to alternative strategies or technologies in the agriculture field to face the environmental and health needs. Synthetic pesticides have been intensively used in agriculture since the second half of 20th century; thus, given the general concern about the environmental and health risk, more sustainable and eco-compatible alternatives are arising. In the present work we focused on the use of natural actives which present antimicrobial activity on different stages of the production process. We developed alginate and carboxymethylcellulose hydrogel beads, which allow the direct and controlled release into the soil of isothiocyanates from Brassicaceae and then a formulation containing actives from Amaryllidaceae and biosurfactants to sanify the fourth range products during washing stage. The beads were characterized by means of ATR-FTIR and DSC measurements, while the loading efficiency and release profile by means of UV-Vis spectrophotometry. The obtained results show that these systems represent a versatile, ecofriendly and non-expensive alternative to biofumigation processes currently employed in agriculture. Liquid formulation based on ungeremine or diacetylicorine in presence of rhamnolipids are studied by means of surface tension measurements, UV-Vis and fluorescence spectroscopy. The results of ecotoxicological assays and microbiological analysis confirm the ecofriendly profile of the formulation which is able to significally reduce the bacterial load. 84 POSTERS 85 ADVANCES IN FOOD AND NUTRITION Antimicrobial profile of sourdough lactic acid bacteria against pathogen bacteria and molds Alina Alexandra Dobre, Elena Mirela Cucu, Laura Adriana Mihai National Research & Development Institute for Food Bioresources, Romania This study aimed to characterize sourdoughs obtained by spontaneous fermentation from Einkorn wheat, corn and rye ecologic flour manufactured following traditional type I sourdough protocol, by their lactic acid bacteria population (LAB). The study also evaluated their antibacterial and antifungal proprieties against pathogen bacteria and spoilage fungi. For this purpose, 20 LAB strains were isolated from which 8 strains were identified to species level using a semi-automated Biolog® Microbial Identification System. The dominant LAB microflora was Lactobacillus bifermentas, Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus casei, Pediococcus acidilactici, Lactobacillus brevis, Lactobacillus coryniformis. The strains were evaluated for their antimicrobial proprieties using agar well diffusion method against pathogen bacteria Staphylococcus aureus, Echerichia coli, Enterococcus faecalis and Bacillus spizizenii and spoilage fungi Aspergillus brasiliensis, Penicillium crysogenum and Mucor racemosus. Most LAB strains showed antibacterial activity on the test microorganisms, especially on B. spizizenii ATCC 6633 but only four LAB strains showed inhibitory effect on the growth and development of fungi. P. chrysogenum was the most susceptible to the action of LAB strains, especially L. plantarum, P. acidilactici, L. coryniformis. Antimicrobial metabolites of these isolates inhibited B. spizizenii and S. aureus, P. crysigenum contaminants from flour, yeast and other sources. Gram-positive bacteria were more susceptible to the action of metabolites produced by lactic acid bacteria strains, E. coli being more resistant to the action lactic bacteria metabolites. According to the results of the study, generally, L. casei, L. brevis and P. acidilactici strains presented a high potential as sourdough starters, in single or mixted starter cultures, because of their antimicrobial activity against bacteria and fungi. Acknowledgments: The results were achieved within the project PN 23 01 02 02 “Research on increasing the shelf life and durability of foods through fermentative processes”, Contract 39 N/16.01.2023 The histaminergic system as a potential target in the treatment of binge eating behaviour in an animal model Zuzanna Koziara1, Emanuela Micioni Di Bonaventura1, Luca Botticelli1, Giacomo Einaudi1, Alessia Costa2, Gustavo Provensi2, Maria Vittoria Micioni Di Bonaventura1, Carlo Cifani1 1University of Camerino, Italy 2University of Florence, Italy Background: Binge eating disorder (BED) is a psychiatric disorder characterized by regularly recurring episodes of binge eating, in which excessive amounts of food are consumed in a short period of time, with a feeling of loss of control over that behaviour. This health issue is often associated with obesity, but so far, anti-obesity medications are not recommended for BED. To date, only one drug (lisdexamfetamine) 86 is approved by FDA as a pharmacotherapy, so there is an urgent need to develop new therapeutic strategies. Much evidence indicates the role of neuronal histamine in the modulation of stress response and feeding behaviour. Therefore, we investigated the changes in the histaminergic (HA) system at the level of gene expression and the effect of HA system targeted compound in an animal model of binge eating behaviour. Methods: Binge eating like behaviour in female Sprague–Dawley rats was induced using food restriction cycles and frustration stress with highly palatable food. After repeated binge eating episodes, rats were sacrificed and selected brain areas were dissected for the RT-PCR gene expression analysis. Furthermore, the effect of an acute and then chronic pharmacological treatment designed to enhance HA neurotransmission was evaluated in the same animal model. Results: Significant alterations in the mRNA expression of HA receptors and enzymes responsible for histamine metabolism were detected in different brain areas of binge eating animals compared to control rats. Moreover, both acute and chronic pharmacological intervention completely blocked binge eating like behaviour and prevented some gene expression alterations. Conclusions: The presented results indicate that the HA system could be involved in binge eating behaviour and might be a new target for pharmacological treatment of BED. Recovery and Green Purification of Methylxanthines and Flavan-3-ols from Cocoa Husk Agro-Waste Laura Morales Erazo, Sonia Sentellas Minguillon, Javier Saurina Purroy Universitat de Barcelona and Institut de Recerca en Nutrició i Seguretat Alimentària, Spain Cocoa husk accounts for ca. 12–20% of the total weight of cocoa beans and is one of the main by-products generated during the pre-roasting and roasting steps [1]. Global cocoa processing generates an estimated 700,000 tons of husk annually, most of which is discarded as waste despite being a rich source of bioactive compounds [2]. Among these, polyphenols such as catechins and procyanidins have been widely associated with antioxidant, anti-inflammatory, and neuroprotective properties, while methylxanthines, including theobromine and caffeine, are known for their stimulant and diuretic effects [3-6], making cocoa husk a promising raw material for sustainable valorization. In this study, a green purification strategy was applied to previously obtained aqueous-ethanol extracts of cocoa husk. A liquid–liquid extraction with food-grade ethyl acetate was initially performed to reduce polar impurities and enrich the target compounds. The resulting semi-purified extracts were subsequently processed using flash chromatography on a reversephase C18 column with an ethanol–water gradient, following green chemistry principles. The antioxidant activity of collected fractions was evaluated by ferric reducing antioxidant power (FRAP) assay. Fractions enriched in flavan-3-ols, particularly epicatechin and catechin, showed the highest antioxidant response, consistent with their known redox behavior [7]. The target compounds were identified and confirmed using high-performance liquid chromatography with diode-array detection (HPLC-DAD) and tandem mass spectrometry (HPLC-MS/MS) based on retention time, UV-Vis spectra, and MS fragmentation patterns in comparison to analytical standards. This study successfully demonstrates a viable and green methodology for the effective recovery of high-value bioactive compounds from cocoa husk, promoting circular economy principles and enhancing the sustainability of agro-industrial processes. [1] J. Panak Balentić, et al., Food Chem. 240, 903 (2018) [2] D.C.G. Okiyama, S.L.B. Navarro, and C.E.C. Rodrigues, Food Res. Int. 99, 1021 (2017) [3] M. Arlorio, et al., Food Chem. 92, 249 (2005) [4] D.C.G. Okiyama, et al., J. Food Sci. Technol. 55, 1692 (2018) [5] E. Martínez-Pinilla, A. Oñatibia-Astibia, and R. Franco, Front. Pharmacol. 6, 30 (2015) [6] C.M. Llerena-Herrera, et al., Heliyon 8, e09601 (2022) [7] P. Zapata-Castillo, et al., Antioxidants 13, 76 (2024) 87 Dietary Potassium Reduction in Fruits and Vegetables: A Study on the Efficacy of Ultrasonic Soaking and Thermal Processing Mojca Korošec1, Alenka Levart1, Simona Vesel1, Gregor Mlinšek2 1University of Ljubljana, Biotechnical Faculty, Slovenia 2University Clinical Center Ljubljana, Slovenia Patients with chronic kidney disease (CKD) face an increased risk of hyperkalaemia, necessitating dietary potassium restriction. However, many potassium-rich foods, particularly fruits and vegetables, are vital components of a healthy diet. This creates a dilemma, as current guidelines often recommend limiting these nutritious options. Our research investigates how various food preparation methods, specifically ultrasonic soaking, can effectively reduce potassium content in fruits and vegetables, thereby expanding safe dietary choices for CKD patients. We analyzed six common fruits and vegetables, using gravimetric analysis and atomic absorption spectrometry (AOAC 999.11) to determine the impact of soaking type (water vs. ultrasound), heat treatment, soaking time, and food piece size on the levels of potassium, sodium, zinc, iron, dry matter, and ash. Our findings indicate that ultrasonic soaking significantly enhances potassium reduction. A 180-minute ultrasonic soak was more effective than water soaking in lowering potassium content to the target level (<200 mg/100 g). Both heat and mechanical treatment (i.e., smaller food pieces) notably influenced leaching efficiency. Heat-treated samples consistently showed lower final potassium levels. For broccoli and carrots, adequate potassium reduction was achieved only through combined soaking and heat treatment. Potatoes required longer soaking times (180 minutes) alongside heat treatment to fall below the threshold. Notably, peaches saw sufficient potassium reduction after just 30 minutes of ultrasonic soaking. Pears and blueberries, however, already had naturally low potassium levels in their raw state, making them suitable for CKD patients without special preparation. We also observed that smaller piece sizes in carrots and potatoes resulted in lower final potassium content. While this study reveals promising trends in potassium reduction through optimized food preparation, a limitation was the single subsample per influencing factor due to the broad scope of foods and variables. Future research should focus on a single food type with multiple subsamples to enable statistical analysis and further confirm these findings, including the influence of natural diversity on elemental content. Evaluation of side-streams from sea buckthorn oil production Cristian Florea, Iulia Elena Susman, Mihaela Multescu, Alina Culetu National Institute of Research & Development for Food Bioresources - IBA Bucharest, Romania Sea buckthorn berries (Hippophae rhamnoides L.) are widely known for their medicinal and nutritional benefits [1, 2]. Extracting oil from sea buckthorn berries produces a significant amount of side-streams, commonly referred to as cake, which is recognized for its high content of bioactive compounds. The objective of this work was to assess and compare the nutritional properties of sea buckthorn cakes generated after two different oil extraction techniques, namely: cold pressing (CP) and supercritical carbon dioxide extraction (SC). In terms of proximate composition, the most notable difference between 88 the two extraction methods was related to the fat content. The sea buckthorn cake obtained through SC extraction had significantly lower fat – about 10 times less – than the cake produced via CP. The CP cake exhibited a darker color, reflected by its lower L* value, and showed a more intense redness, indicated by a higher a* value, compared to the SC cake. The chromaticity towards yellow (b*) displayed comparable values for both cakes. The CP cake exhibited a higher concentration of total phenolics (15 mg gallic acid equivalent/g d.m.) and flavonoids (4.9 mg quercetin/g d.m.) compared to the SC cake. Additionally, antioxidant capacity, as assessed through four complementary methods, showed higher values for CP than SC. The comparative antioxidant capacity values (expressed as mg Trolox/g d.m.) for CP and SC cakes were as follows for each investigated assay: 9.6 vs. 6.3 (DPPH), 19.9 vs. 14.0 (FRAP), 45.0 vs. 31.9 (ABTS), and 36.8 vs. 24.8 (CUPRAC). The CP and SC sea buckthorn cakes are characterized by relatively high content of polyphenols, flavonoids and antioxidants, reinforcing their potential as valuable raw materials for enhancing the nutritional profile of food products. The findings highlight opportunities for using these side-streams from the sea buckthorn oil industry in food development. To improve their integration into food formulations, efforts should concentrate on optimizing sensory attributes, addressing atypical aromas and flavors, and evaluating consumer acceptance to ensure market viability. This work was supported by a grant of the Ministry of Research, Innovation and Digitization, CNCS/CCCDI UEFISCDI, project number ERANET-JPI-HDHL-Up4Food, within PNCDI IV (contract no. 31/2024). References: [1]. Zulfiqar R., Fatima K., Ul-Haq, I., et al., Hum. Nutr. Metab 41, 200321 (2025) [2]. Singh B., Oberoi S. and Kaur A. Int. J. Food Sci. Technol 59, 6642–6658 (2024) Exploring the amino acid profile of various legumes for optimal nutrition Mihaela Multescu, Iulia Elena Susman, Cristian Florea, Alina Culetu National Institute of Research & Development for Food Bioresources - IBA Bucharest, Romania Legumes have an important role in promoting healthy and sustainable human diets due to their high content in protein, fiber, and bioactive compounds. The nutritional quality of food proteins is largely determined by their essential amino acids composition and digestibility. While the amino acid profile of legumes varies across different types, legume proteins are generally deficient in sulfur-containing amino acids. However, they are abundant in lysine, making them complementary to cereal proteins in a balanced diet. This study aimed to analyze the amino acid profile and assess the nutritional quality of protein in various legumes, including lentils (red, green, yellow, brown, black), beans (mung, pinto, black, kidney), chickpea, soy, and lupin. Amino acid analysis was conducted using a highperformance liquid chromatography system with an automatic pre-column derivatization method [1]. The results are expressed as g/100 g protein. Glutamic acid (15.2-27.6%) and aspartic acid (10.4-13.9%) were the most abundant amino acids, while methionine (0.8-1.6%) and cysteine (0.6-1.5%) had the lowest concentrations. In all the legume samples analyzed, the essential amino acids leucine, valine, isoleucine, threonine, histidine, lysine, and aromatic amino acids (phenylalanine + tyrosine) met the FAO recommended requirements. Chickpea and soy fulfilled the FAO requirements for sulfur-containing amino acids (>2.5%), whereas green and brown lentils had the lowest essential amino acids content. According to the amino acid scores (AAS), soy, black bean, kidney bean, and mung mean did not have any limiting amino acids. Protein digestibility corrected amino acid score (PDCAAS) values indicated that soy, chickpea, and mung bean generally exhibited high protein quality. Based on the essential amino acid index (EAAI), green and brown lentils had low protein quality (EAAI values below 75), while all the other legumes samples displayed moderate protein quality (EAAI between 75 and 86). Combining legumes or blending them with cereals are essential to develop innovative food products that meet the evolving preferences of modern consumer. 89 This research was achieved through the Core Program, with the support of the Ministry of Research, Innovation, and Digitization (MCID) (contract 39N/2023; project PN 23 01 01 02). References: [1] Multescu, M., Culetu, A. and Susman, I.E. Foods 13, 3528 (2024) Co-creation from University-based Living Labs: the case of the FUSTO Programme Angelo Riccaboni1, Patrizia Marti1, Marco Romi1, Cristiana Tozzi1, Francesca Gagliardi1, Claudia Zoani2 1University of Siena, Italy 2Italian National Agency for New Technologies, Energy and Sustainable Economic Development (ENEA), Italy FUSTO (From Soilless to Table and Beyond) is an interdisciplinary innovation and learning program aimed at advancing sustainability and innovation in agri-food systems, operating within the framework of national programs such as AGRITECH (Spoke 9) and METROFOOD-IT, funded by the Italian NRRP. FUSTO connects the Santa Chiara Lab of the University of Siena, a certified Living Lab under the European Network of Living Labs (ENoLL), and Siena Food Lab Foundation, a Living Lab created by the University of Siena and the Monte dei Paschi Foundation, whose partners include seventy agricultural enterprises, a secondary school and all local farmers associations. In this way, a wide ecosystem connecting academia, industry, and local communities was developed, integrating in a perspective of co-creation scientific research, technological development, and stakeholder engagement. FUSTO addresses the entire value chain—from primary production to processing, packaging, consumption, and circular reusesupporting a wide array of research and technology transfer activities, concerning: - Soilless cultivation research using advanced hydroponic and aeroponic systems to optimize plant growth in resourceefficient environments. - Sustainability assessment and ESG reporting through the ESG4Agri platform and LCA software (SimaPro), helping farmers and agrifood companies to evaluate and communicate their environmental and social performance. - AGRIHUB is an innovative platform that integrates a comprehensive database of sustainability indicators across the agri-food supply chain, collected from both primary and secondary sources. Through an interactive dashboard, users can easily and intuitively visualize the data using charts and tables. - Geographical and varietal traceability of agri-food products using isotopic, genetic, and chemometric analyses to ensure quality, safety, and authenticity. - Consumer behavior analysis using virtual reality, focus groups, and participatory methods to explore perceptions and choices related to food sustainability and innovation. - Food transformation and design in the Kitchen Lab, integrating 3D food printing, mold design, and dietary personalization aligned with Mediterranean diet principles. - Sustainable packaging innovation, developing biodegradable and functional materials from agri-food waste with antimicrobial and antioxidant properties. Through its Living Lab methodology, Fusto fosters co-creation among diverse stakeholders, ensuring that innovations are grounded in real-world needs and aligned with the Sustainable Development Goals (SDGs), particularly food security (SDG 2), sustainable consumption and production (SDG 12), and climate action (SDG 13). This dynamic ecosystem exemplifies how university-based Living Labs can cooperate among them and serve as effective platforms for systemic innovation, bridging research and practice to drive sustainable transformation in the agri-food sector. Acknowledgment: This work was supported by METROFOOD-IT (CLP - IR0000033) and Agritech (CUP CN00000022) projects, funded by the European Union - NextGenerationEU, PNRR - Mission 4 “Education and Research”. 96 Real-time AI-based monitoring system for microbiological safety of drinking water Michela Zampano1, Rosario Schiano Lo Moriello1, Francesco Bonavolontà1, Giorgio De Alteriis1, Stefano Batistini2, Tommaso Pellino2, Marco Guida1, Federica Carraturo1, Raffaele Velotta1, Edvige Gambino3, Bartolomeo Della Ventura1, Elena Andolfi2 1Universita degli studi di Napoli Federico ll, Italy 2Opus Automazione S.P.A, Italy 3CESMA, Italy The WAFER (WAter SaFEty Plan Automatic MonitoRing System) project aims to transform microbiological water quality monitoring by developing an intelligent, fully automated platform for real-time pathogen detection. The project addresses the urgent requirement for decentralized, continuous control of drinking water safety and is aligned with evolving regulatory frameworks, such as EU Directive (EU) 2020/2184. WAFER integrates state-of-the-art biosensors, artificial intelligence and Internet of Things (IoT) technologies into a mobile, energy-autonomous unit designed for frequent, unattended operation. The system performs a fully automated workflow, including sampling, fluid treatment and biosensorbased analysis, capable of detecting contamination events earlier and more efficiently than conventional, laboratory-based methods such as PCR, which are slow, resource-intensive and restricted to centralised facilities. At the core of WAFER lies an AI-driven module that processes environmental and biological data in real time using adaptive machine learning models. This enables immediate alert generation, predictive diagnostics, and continuous model refinement. The biosensors exhibit high sensitivity and selectivity towards key microbial indicators, including Escherichia coli and intestinal enterococci, which are classified as fundamental parameters under EU and national regulations. Additionally, the system is designed to detect a broader range of pathogens, such as enteric viruses and AMR-related bacteria, enhancing its applicability across diverse health risk scenarios. The unit is designed for robust outdoor use and includes edge computing capabilities for low-latency data processing. Secure transmission protocols ensure integration with cloud platforms, enabling visualization, storage, and correlation with broader water safety and epidemiological data systems. WAFER supports remote diagnostics and over-the-air updates, making it suitable for large-scale network deployment with minimal operational burden. WAFER strengthens the resilience of water infrastructures by enabling real-time, autonomous monitoring, enhancing early warning capabilities, and supporting long-term environmental and public health strategies. Deploying WAFER contributes directly to Sustainable Development Goals (SDGs) related to clean water, health, and innovation, positioning it as a pioneering solution in the field of smart environmental monitoring. This work was supported by Italian Ministry for Industry and Made in Italy (MIMIT) program “SCOPERTA IMPRENDITORIALE” within the framework of the project WAFER, beneficiary company Opus Automazione S.p.A. 97 METROLOGY IN FOOD METROFOOD-RI integrated analytical services: Phenol determination in agro-industrial by-products and derived formulations Maria Z. Tsimidou1, Nikolaos Nenadis1, Aggeliki Kalogeropoulou1, Claudia Zoani2 1Aristotle University of Thessaloniki - AUTH , Greece 2ENEA CR CASACCIA, Department for Sustainability, Biotechnologies and Agroindustry Division (SSPTBIOAG), Italy METROFOOD-RI is a distributed European research infrastructure aimed at promoting scientific excellence in the field of metrology in food and nutrition (www.metrofood.eu). Considering its role as serviceoriented entity, within the frame of METROFOOD-EPI project (HEu, GA 101130162), competitive open calls were launched to provide transnational access (TNA) with physical and remote contact to users, who were selected on the basis of excellence of their proposals. For the accomplishment of the call focused on food production and integrated analytical services, a pre-defined set of grouped combinatorial analytical services and food processing technologies involving multiple service providers was made available. The selected applicants provided the raw materials and asked for the development of specific formulations which would incorporate functional ingredients extracted from the former. Compositional analysis for both by-products and formulations was assigned to various laboratories upon pre-declared expertise. Aristotle University of Thessaloniki (AUTh) analytical facilities were involved in the analysis of polar and non-polar phenolic compounds using extraction and analytical protocols employing chromatographic techniques (NP-HPLC-DAD-FLD and RP-UHPLC-DAD-FLD). This work presents the “service section” provided by AUTh facilities with focus on metrological aspects for the selection of the fit for the purpose protocols. Acknowledgements: Research was funded by the METROFOOD-EPI project (HEu, GA No 101130162). The project ‘Dissemination and exploitation of the outcomes of the Laboratory of Food Chemistry and Technology’ (AUTh Research Committee code 96677) is acknowledged for financial support of MZT participation. The Impact of Equipment Conformity Assessment in Ensuring the validity of Food Testing: A Comparison of Laboratory Practices Makhmud Khayrullaev, Normurod Boymatov Uzbek Center Accreditation, Uzbekistan Ensuring food safety is impossible without the reliability of measurement results and laboratory tests. Modern approaches to quality control in the food chain require strict adherence to the principles of metrological traceability, conformity assessment of measuring instruments and harmonization of laboratory procedures with international standards. The ISO/IEC 17025 standard sets out key requirements 98 for the competence of testing and calibration laboratories, covering both technical and management aspects. In addition, the ISO 10012 standard regulates the requirements for the measurement management system and the operation of measuring equipment, ensuring the integration of metrological functions into the overall quality management system. Certified reference materials (CRMs), the production of which is regulated by ISO 17034, play an important role in ensuring the reliability of measurements. They are used as a reference material for the verification of analytical methods, as well as within the framework of the participation of laboratories in proficiency testing (PT) programs according to ISO/IEC 17043. The combined use of these standards improves reproducibility indicators, as well as ensures metrological traceability and reliability of analytical results. In this study, a comparative analysis of metrological practices was conducted in 24 testing laboratories in various regions of Uzbekistan, including more than 10 accredited according to ISO/IEC 17025. The study examined methods for determining pH in food products using various types of samples: certified reference materials (CRMs) and artifact materials, the homogeneity and stability of which were assessed in accordance with the requirements of ISO/IEC 13528 in an accredited laboratory. The obtained data indicate that laboratories accredited in accordance with international standards achieve a higher degree of metrological traceability due to the systematic application of calibration procedures, intermediate checking and regular participation in interlaboratory comparison tests. At the same time, non-accredited laboratories of Uzbekistan continue to apply verification and metrological certification procedures inherited from the Soviet regulatory system, which is also aimed at ensuring reliability, but does not always meet the requirements of international practice. The results of the study confirm that the presence of an established metrological infrastructure and mechanisms for assessing the conformity of measuring instruments has a decisive impact on the reliability of control of food safety indicators. Note: The analysis is in line with the direction of the 8th IMEKOFOODS Conference (TC23), which pays special attention to the international harmonization of measurement methods in the food industry, as well as traceability and confirmation of the reliability of analytical data. Metal impurities in food additives: Do they contribute to dietary metal exposure? Céleste Vansnick, Karlien Cheyns, Nadia Waegeneers Sciensano, Belgium Toxic elements such as lead (Pb), cadmium (Cd), arsenic (As) and mercury (Hg) may enter the human food chain through the soil-plant-animal pathway, as well as through industrial processes, including their presence as impurities in food additives (FAs). Commission Regulation (EU) No 231/2012 outlines the specifications for FAs, including impurity limits. However, a preliminary intake assessment based on EFSA exposure estimates for the Belgian population indicated that, if these metals were present in FAs at concentrations approaching the regulatory limits, the resulting dietary intake could exceed general dietary exposure estimates by a factor of 2 to 12. This suggests that actual concentrations are substantially lower than specified limits and/or that current exposure models may underestimate intake from foods containing FAs. The present study aims to evaluate the exposure of the Belgian population to Pb, Cd, As, and Hg due to FA intake. The objectives are threefold: (i) to quantify the occurrence of metal impurities in selected FAs, (ii) to assess dietary exposure to these metals specifically from FA consumption, and (iii) to compare this exposure with current general dietary exposure estimates and conduct a risk assessment for the Belgian population. A prioritisation approach, considering both FA characteristics and Belgian consumption patterns, was used to select 17 food additives for analysis: E120, E170, E300, E301, E330, E401, E407, E410, E412, E415, E422, E440, E461, E500, E508, E509, and E516. Analytical methods to determine the concentration of metals in these different matrices were validated using inductively coupled plasma – mass spectrometry (ICP-MS) and Direct mercury analysis (DMA). This resulted in 99 expanded measurement uncertainties ranging from 9-33%. At least five samples of each selected additive were analysed, of which metal concentrations will be presented. The results of these analyses will be used to conduct a targeted sampling and analysis of FA-containing foods that may contribute to metal exposure. Based on these data, a re-evaluation of the exposure of the Belgian population to Pb, Cd, As and Hg will clarify whether FAs significantly contribute to metal exposure. Acknowledgement: This research was funded by the Belgian Federal Public Service of Health, Food Chain Safety and Environment, project RF 23/23: Metalfood@. We would like to thank Iris Wyns for her valuable contribution to the project. Standardizing Plant Pathogen Detection using dPCR as a Reference Method for Xylella fastidiosa Alexandra Bogožalec Košir, Špela Alič, Manca Pirc, Neža Turnšek, Tanja Dreo, Mojca Milavec National Institute of Biology, Slovenia Reliable detection of plant pathogens is critical for ensuring biosecurity and food safety across the agricultural value chain. This study addresses two major aspects influencing diagnostic accuracy for Xylella fastidiosa, a quarantine pathogen of high regulatory concern in the EU: (i) the influence of DNA extraction methods, and (ii) the application of digital PCR (dPCR) for metrologically traceable measurements. Four different DNA extraction protocols, QuickPickTM SML Plant DNA Kit, Modified DNeasy Mericon Food Standard Protocol, Maxwell® RSC PureFood GMO and Authentication Kit, CTAB, were evaluated across diverse X. fastidiosa subspecies and plant matrices to determine their yield, repeatability, and matrix effects. Results showed significant variability among methods, particularly when processing complex plant samples such as olive tree (Olea europaea) leaves. This emphasizes the critical role of selecting the appropriate extraction protocol. In parallel, we developed and validated a dPCR method for X. fastidiosa quantification, following ISO 20395:2019 and EPPO PM 7/24 (3) standards. Three levels of complexity, synthetic targets, bacterial genomic DNA, and plant-spiked genomic DN, were analysed on four different dPCR platforms, QX200 (BioRad), Naica (Stilla Technologies), QIAcuity (Qiagen) and Absolute Q (ThermoFisher Scientific). The method demonstrated high sensitivity (LOD ~ 9 copies/ reaction), excellent intermediate precision (CV < 8 %), and robust inter-platform reproducibility (CV < 17 %). This work highlights the necessity of rigorous evaluation of extraction protocol performance in plant pathogen diagnostics and supports the role of dPCR as a potential reference measurement procedure for regulatory and research laboratories. Development and validation of control materials for molecular detection of Listeria monocytogenes in milk Pia Marinček1, Darja Kušar2, Jana Avberšek2, Mojca Milavec1, Alexandra Bogožalec Košir1 1National Institute of Biology, Slovenia 2University of Ljubljana, Veterinary Faculty, Slovenia Listeria monocytogenes is a foodborne pathogen that poses a severe threat to vulnerable populations. It can proliferate at refrigeration temperatures and persist in ready-to-eat foods such as dairy products. The most effective strategy to prevent serious outbreaks is the implementation of a well-designed, 100 standardized monitoring system. Currently, global standards for the detection and quantification of L. monocytogenes rely on microbiological culture methods that require several days to yield results. However, recent advancements in molecular biology, particularly the development of nucleic acid amplification-based methods such as polymerase chain reaction (PCR), have enabled rapid and accurate detection and quantification of microorganisms. Compared to traditional culture methods, PCR technology delivers results within a single day. Digital PCR (dPCR) enables absolute quantification traceable to the SI system of units, making it a promising tool for the future of standardized microbial monitoring. Besides precisely described and evaluated measurement procedures, metrological frameworks require fit-for-use reference materials to ensure traceability and harmonization of measurements across laboratories. In this study, we developed control materials for the detection and quantification of L. monocytogenes in milk and evaluated them using a dPCR assay targeting a speciesspecific, single-copy gene. We spiked milk samples with L. monocytogenes suspensions to create two materials, one with a high concentration estimated at 5E6 cells/mL and one with a lower concentration of approximately 5E4 cells/mL. A positive control sample consisted of genomic DNA that was extracted from a bacterial suspension estimated at 1E7 cells/mL and eluted in a stabilizing buffer. All of the prepared materials were assessed for homogeneity and stability. We evaluated the selected dPCR measurement procedure and calculated its measurement uncertainty associated with repeatability, intermediate precision and variation in the volume of partitions. The produced control materials were homogeneous and stable in the short term. However, after two months of storage, a substantial decrease in bacterial concentration could be noticed for milk materials with the lower bacterial concentration (5E4 cells/ mL), while the positive control remained stable. Our findings underscore the importance of studies on control materials that closely mimic real-life samples, highlighting the challenges with developing stable materials in complex matrices such as milk. They represent a step towards the integration of standardized molecular methods into the legislative monitoring of foodborne pathogens that would benefit public health with fast, precise and accurate detection using the dPCR technology. Grain moisture determination. Using of standard addition method Olena Shcherbakova, Serhii Kulyk, Oleksii Stennik, Oleksandr Melnykov SE Ukrmetrteststandart, Ukraine Grain moisture content is a critical factor for storage optimizing and establishing a fair price for grain. However, according to the data of inter-laboratory comparisons, the moisture determination results obtained by main Loss-on-Drying (LoD) methods (ISO 712, ISO 6540) in different agricultural laboratories differ significantly. In particular, the range of variation reaches 1.5% in Ukrainian laboratories. At the same time, expanded uncertainty of a single result is estimated at only 0.3% (ISO 712). In our opinion, the reason for this variation in results may be the expressed influence of the equipment used for LoD methods execution. For example, the loss of moisture during grain grinding, insufficient efficiency of the drying oven, etc. can lead to an underestimation of moisture content. In our study, we tried to verify the correctness of the LoD method using the standard addition method. A precisely defined amount of water was added to wheat grain sample with a known moisture content. Grain samples were encapsulated in a sealed package. After stabilizing for approximately 1 month, the moisture content of the moistened sample was determined according to ISO 712. It was expected that if the measurement procedure was not implemented correctly, the result would differ significantly from the theoretically calculated value. According to obtained data, the use of a knife mill without cooling the working chamber leads to significant (up to 0.3 %) decrease in the moisture measurement result compared to the calculated one. At the same time, significant result underestimation wasn’t observed for the 3310 Perten disc mill. Other equipment (oven, desiccator, balance) remained unchanged. Comparison of the results obtained 101 with different mills showed that the use of knife mills without cooling leads to a systematic significant results underestimation up to 0.7%. If knife mill is equipped by cooling system, as in the case of the KN 295 Knifetec, no underestimation was observed. For cases where the difference between theoretical and experimental moisture content values is insignificant, the SD of these differences was estimated as 0.04 %. The results of the study show the importance of assessing equipment influence on the LoD method results during moisture content determination. The standard addition method can be used to assess this influence. The results can be used to discuss possible changes to ISO 712 and ISO 6540. Also, the data obtained can be used to assess the characterization uncertainty of CRM of grain moisture content. Metrology for the phosphorus content measurements in vegetable oils in support of European and international food safety Dmytro Chupis SE Ukrmetrteststandart (UMTS), Ukraine Metrology for the phosphorus content measurements in vegetable oils in support of European and international food safety and trade Crude vegetable oils usually have impurities (phospholipids, as a specific class of phosphorus-containing compounds, FFAs, metals, etc.) which during the refining process may cause damage to the vegetable oil’s stability, color, odor and taste, so that their removal is necessary. Having important physiological properties, phospholipids at the same time complicate the subsequent stages of vegetable oil processing and negatively affect the technical and economic indicators of refining and hydration in general. The presence of phospholipids in the subsequent stages of refining may damage the vegetable oil color, odor and taste. Phosphorus content in oil is a crucial quality parameter; thus, accurate and fast phosphorus content measurement supports fair trade in the vegetable oil market. To verify and calibrate phosphorus analyzers and ensure their traceability, it is necessary to use matrix phosphorus content CRM. We aim to create matrix reference materials for phosphorus content in vegetable oils in support of national and European and international food safety and trade. Many stakeholders are interested in this topic. The work can be performed by following steps: – preliminary investigation (literature overview, measurements); – creation of candidate RMs; – pilot comparison; – key comparison; – CMC claims; – creation of the CRMs. UMTS has two years experience in this field and permanently collaborate with oil manufacturers. Unrefined oils with different phosphorus content are provided by oil manufacturers on permanent basis. UMTS produce uniform batches of unrefined oils and study the homogeneity and stability for batches are studying. Currently, the preliminary studies for more than 15 oil batches are performed. As part of this effort, we have already obtained comparative measurement results using various analytical methods, including gravimetry, photometry, and X-ray fluorescence (XRF). Current data confirm the comparability of these methods with the expected target measurement uncertainty. Developing phosphorus content matrix reference materials will make possible to provide metrologically traceable measurements. 102 FOOD PACKAGING What is the future of FCM after the ban on single-use plastic articles? A deep dive into alternative materials in Belgium Mélanie Di Mario Sciensano, Belgium Since the ban on single-use plastic articles in Europe, the food contact material (FCM) industry has been forced to move to more sustainable alternatives. At the Belgian level, the Federation of Belgian Food Industry (FEVIA) states that all food packaging must be reusable, recyclable, or compostable by 2025. One way to tackle the waste problem is to introduce “environmentally friendly FCM”, which must also be safe for consumers. Many options are currently available in the market. However, the market share, trends, and consumer preferences remain unclear. Therefore, this study aims to provide an overview of the Belgian FCM market, as well as the substitute materials available and their applications. The market analysis embraced an integrated web-based approach, diving into fifty-two diverse sources. These included e-shops dedicated to offering innovative alternatives to disposable plastics and showcasing environmentally friendly options, along with websites that inspire creativity through homemade food contact materials. The initial screening revealed a total of 10,523 articles. After a thorough data cleaning process, we achieved a consistent dataset consisting of 2,688 unique entries, divided into fifteen material categories and seven utilisation classes. Paper and board were the most commonly used material categories intended to replace single-use plastic articles, comprising 37% of the entries, followed by bagasse, which accounted for 9%. The most frequent usage categories were takeaway and food serving, representing 44.4% and 22.8% of the entries. Enhanced untargeted method to detect organic and inorganic contaminants in biobased and recycled food contact materials Alessia Turina1, Francesco Romaniello2, Matteo Berruto2, Consolato Schiavone2, Claudio Medana1, Andrea Mario Rossi2, Chiara Portesi2 1Univerità degli Studi di Torino, Italy 2Istituto Nazionale di Ricerca Metrologica, Italy Food packaging plays a vital role in maintaining food quality and safety1. Driven by the need to reduce waste and environmental impact, the development and use of bio-based, biodegradable, and recycled materials as sustainable alternatives to conventional plastics and paper2 are increasing. In parallel, the monitoring of novel contaminants potentially present in these emerging materials, including those originating from recycling processes, is needed to ensure their compliance with legislation and their safety. To address this critical need, a validated harmonized untargeted method for the comprehensive identification of both organic and inorganic contaminants is essential for characterizing the contaminant profile of these innovative food contact materials, particularly given the current lack of standardized 103 analytical approaches at the European level. This work has been developed in the framework of the 23IND13 ScreenFoodproject and aims to develop and harmonize untargeted methods using Liquid Chromatography-Mass Spectrometry (LC-MS) system for organic contaminants and Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) for inorganic contaminants. Various types of packaging, both recycled and non-recycled, including cardboard, plastic, and biobased materials such as bagasse, he, were analyzed. The first part of the work was devoted to the optimization of the extraction methods both for organics and inorganics. Existing guidelines and protocols were optimized for the different types of packaging, each of which posed specific challenges based on its chemical nature, shape and final intended use and further analysis (organic and/or inorganics). Afterwards, non-targeted analysis, for identifying organic contaminants in food contact material samples, was performed using LC-MS system Thermo Fisher Q Exactive Hybrid Quadrupole-Orbitrap with ESI ionization source, in both positive and negative modes. The acquisition mode used was full scan-data dependent acquisition (DDA). The untargeted LC-MS method enabled the detection of different organic contaminant classes such as PFAS, phthalates, bisphenols, photo-initiators, primary aromatic amines and mineral oils. The ICP-MS system used for inorganic contaminants detection was Thermo Fisher XR element. ICP-MS analysis revealed the presence of heavy metals in food packaging samples, with Iron (Fe), Manganese (Mn), Copper (Cu), and Zinc (Zn) identified as the most abundant elements.The proposed strategy successfully characterized the contaminant profile of the food packaging materials sampled by Screen food partners. This approach has proved to be a reliable testing method for identifying the main contaminants in emerging recycled, sustainable, and biodegradable food contact materials, to assess and ensure the safety of food contact materials. Acknowledgments: The present work has been supported by the project project 23IND13 ScreenFood, which has received funding from the European Partnership on Metrology, , co-financed by the European Union’s Horizon Europe Research and Innovation. Microplastics on the Surface of Packaged Chicken Meat: Method Optimization for Detection, Isolation, and Characterization Tina Šaula1, Živa Kolenc1, Sonja Smole Možina1, Manca Kovač Viršek2, Anja Klančnik1 1University of Ljubljana, Biotechnical Faculty, Slovenia 2National Institute of Biology, Slovenia Microplastics are increasingly recognized as a food safety issue as they are widely distributed in the environment and can carry contaminants on their surface. In the context of food production, microplastics can come from various sources, such as processing equipment, packaging material or the air, and can end up on the surface of food during handling and processing. The aim of this study is to assess the presence of microplastics on chicken meat by optimizing meat surface sampling, chemical digestion of the meat, and the filtration process prior to isolating and characterizing microplastics from the filters. Chicken meat products of different producers and packaged in various packaging materials were sampled under sterile and dust-free conditions. A 50 g portion was taken from the outer 10 mm of the meat surface and then digested in 10% potassium hydroxide at 36 °C for 48 hours at 80 rpm. The resulting suspensions were filtered through 10 µm nylon filters. Suspected microplastics were isolated manually using pointed tweezers and visually examined under a stereomicroscope according to their morphological characteristics such as size, shape and color. The particles were then analyzed using Fourier transform infrared spectroscopy (FTIR) to identify the polymer type. Microplastics were successfully isolated from all meat samples with the number of particles per sample ranging from 1 to 104 6. Most of them were classified as fibers or films, ranging in size from 0.2 to 2.2 mm, and were mostly translucent or green, matching the color of some of the packaging materials. FTIR analysis confirmed that some of the isolated particles matched the polymer composition of packaging material, indicating it as one of possible routes of contamination. However, microplastics of other polymer types were also detected, indicating other routes of contamination, probably occurring during meat processing. We have optimized and applied a method for the detection and isolation of microplastics from chicken meat. The results indicate that microplastics from meat packaging and also other sources are present on the surface of chicken meat and thus present a direct (physical, chemical) and indirect (microbiological) food contamination threat. These results emphasize the need for further research into the role of microplastics in food contamination and for food safety assurance within the food supply chain. Acknowledgements: This research was supported by the Slovenian Research and Innovation Agency (ARIS) under the project MR-58148, J4-4548 and P4-0116. Poly (lactic acid)–Poly (L-lactide-co-ethylene adipate) Hybrid Films: A Promising Approach for Sustainable Food Packaging Athira John1, Klementina Pušnik Črešnar1, Johan Stanley2, Sabina Vohl1, Damjan Makuc3, Dimitrios Bikiaris2, Lidija Fras Zemljič1 1University of Maribor, Slovenia 2Aristotle University of Thessaloniki, Greece 3Slovenian NMR Centre, National Institute of Chemistry, Slovenia Poly(lactic acid) (PLA) is extensively employed across various industrial sectors due to its biodegradability; however, inherent limitations such as low crystallinity adversely impact its mechanical strength, thermal stability, and chemical resistance. Its susceptibility to hydrolysis and water absorption, are particularly problematic, reducing long-term durability. To mitigate these issues, this research investigates the formulation of hybrid films through blending PLA with poly (l-lactide-co-ethylene adipate) (pLEA) block copolymers. The study specifically aims to enhance crystallinity, mechanical performance, and chemical durability of PLA. Advanced analytical techniques, including Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Polarized Light Microscopy (PLM), Differential Scanning Calorimetry (DSC), and Thermogravimetric Analysis (TGA), were utilized to comprehensively characterize the structure-property relationships within the blends. Through systematic optimization of blend ratios and processing conditions, notable enhancements were achieved compared to pure PLA. Remarkably, films containing 30 wt% pLEA (97.5/2.5) and 70 wt% PLA, fabricated via rapid cooling and a brief annealing period of just 5 minutes, transitioned from amorphous to highly crystalline structures. This resulted in approximately a 20 MPa increase in tensile strength, greater stiffness, reduced surface wettability, and improved resistance to chemical degradation. Significantly, these optimized films exhibited enhanced hydrophobicity, evidenced by a reduction of approximately 15° in contact angle and a roughly 7 mN/m decrease in the polar component of surface free energy. Such improvements suggest increased resistance to moisture and polar solvents, thus enabling superior barrier performance and chemical resilience. In conclusion, the developed strategy of copolymer blending effectively addresses inherent limitations of PLA, presenting a viable pathway for creating high-performance biodegradable materials suitable for demanding industrial applications[1]. Reference: [1] A. John, K.P. Črešnar, J. Stanley, S. Vohl, D. Makuc, D.N. Bikiaris, L.F. Zemljič, Polym Test 143,108707 (2025) 105 FOOD PROCESSING Food Processing Workshops in the RURBANIVE Project: Helping Local Producers Shorten the Supply Chain Lenka Kourimska, Marie Simpachova Pechrova Czech University of Life Sciences Prague, Czech Republic The RURBANIVE project (Rural-uRBAN synergies emerged in an immersIVE innovation ecosystem) aims to improve life in local rural communities through social and technological innovation. One of its key objectives is to enhance logistics and shorten the value chain. As part of this effort, a series of workshops and online courses were organized for a variety of stakeholders. These activities focused on modern food processing technologies that can help farmers and local producers shorten the journey of food products from production to direct sale on the farm or at local markets. In 2025, four workshops were held at the CZU Food Pavilion, covering meat, milk, bakery, and beer technologies. A total of 57 stakeholders participated, including representatives from public authorities, academia, research institutions, and agri-food producers. The workshops addressed practical topics such as Haloumi cheese production, enriched bread and bakery products, beef aging techniques, and beer brewing. According to postworkshop surveys, 88% (bakery), 100% (milk), 94% (meat), and 54% (beer) of participants stated they would definitely or probably apply the knowledge and skills gained in practice. Based on the workshops, a series of online courses is being developed to provide further guidance on food processing, labelling, certification, and inspection. These activities aim to foster co-creation among stakeholders, strengthen communication between academia and producers, and accelerate the implementation of scientific knowledge into practice. Acknowledgement: This work was supported by the RURBANIVE project, which is co-funded by the European Union under Grant Agreement No. 101136597. Antibacterial Potency of Slovenian Honeys against Foodborne Pathogens Tina Šaula1, Martina Miceva1, Tjaša Damijan1, Mateja Šuštar1, Urška Ratajc2, Sonja Smole Možina1 1University of Ljubljana, Biotechnical Faculty, Slovenia 2Slovenian Beekeepers’ Association, Slovenia Honey has been used for nutritional and also therapeutic purposes since ancient times and has been found also to exhibit antimicrobial activity against broad spectrum of microorganisms. The antimicrobial properties of honey depend on several factors, including phenolic compounds, enzymatic production and degradartion of hydrogen peroxide, high osmolarity and low pH level. Beside this, the botanical origin, classified into the nectar/floral (light) and mannin/forest (dark) honey, and geographical origin of honey are also decisive. The antimicrobial activity also depends on the species of bees, environmental, seasonal, and climatic conditions that affect the composition of the honey, as well as the time of honey collection and subsequent handling. The spectrum of antimicrobial activity of 85 honey samples of defined botanical origin (multifloral, linden, acacia, fir, chestnut and forest) from all Slovenian regions 112 sensitive and rapid GC-MS/MS method combined with the QuEChERS technique with modifications was developed for the simultaneous determination of 4 PAHs in cereal-based products (semolina, biscuits, pasta, breakfast cereals). QuEChERS extraction was performed with citrate salts and acetonitrile, and purification was realized by freezing and solid-phase extraction (d-SPE) with Z-Sep+ sorbent in different amounts and combinations with magnesium sulfate. Analysis of 4 PAHs was performed using the internal standard method by selected reaction monitoring mode using a GC-MS/MS. The method was validated following the requirements established by the European Commission Regulation 2021/808. Mean recoveries for the 4 PAHs were determined for intra-day, inter-day, and intermediate precision conditions, by fortifying a blank sample at 1, 2, and 3 µg/kg concentrations and ranged between 74-115%, 86-108%, and 92-113%. Intra-day precision (RSD) ranged between 1.1-11.7%, 1.5-8.7%, and 1.5-6.3%, while interday precision values were 2.1-20.2%, 1.7-10.9%, and 1.1-11.4% for the three spiking levels. Linearities were checked on calibration curves plotted on standard solutions (0.5-3.5 μg/L) and on matrices (1-7 μg/ kg). The method showed good linearities (R²≥0.99 for all analytes). LODs ranged from 0.01 to 0.21 μg/kg, and LOQs from 0.05 to 0.69 μg/kg. Trueness was verified for the 4 PAHs, separately and as the sum of 4 PAHs, by participating in a PT test of a corn semolina matrix, on three concentrations: < LOQ, 2.49 – 2.79 (∑4PAH=10.24) µg/kg, 6.93 – 7.75 (∑4PAH=29.84) µg/kg), obtaining average z-scores of: BaA, z= 0.42; Chr, z= 0.80; BbF, z= 0.64; BaP, z= 1.24; ∑4HAP, z= 0.66. The developed and validated method was applied to matrices like cereal flours (4), breakfast cereals (1), fresh bread (2), toast/crumbs (2), semolina (2), and biscuits for young children (7). None of the 4 PAHs were quantified in any sample. The proposed method met the criteria of Regulation 2021/808 and can be used to quantify the 4 PAHs in processed cereal products. Fungal and Yeast Contaminants in Dried Fruits: Implications for Food Quality and Safety Tatjana Radovanović Vukajlović, Melitta Buzás, Shanique Walker, Ayşegül Kayaaltı, Mitja Martelanc, Anej Colja, Anuša Skušek, Katerina Mukhina, Laura Smolnikar Štrukelj, Valentina Ratoša, Sara Gregorc, Branka Mozetič Vodopivec, Lorena Butinar High content, low release: Exploring TiO2 nanoparticle migration from food packaging using APEX™-single particle ICP-MS/MS Isabel Bastardo-Fernandez1, Rachida Chekri1, Johanna Noireaux2, Caroline Oster2, Valentin Thoury3, Paola Fisicaro2, Petru Jitaru4 1Laboratory for Food Safety, French Agency for Food, Environmental and Occupational Health and Safety (ANSES), Université Paris-Est, Maisons-Alfort, France 2National Laboratory of Metrology and Testing (LNE), Paris, France 3Industrial Technical Centre for Plastics and Composites (IPC), Alençon, France 4French Agency for Food, Environmental and Occupational Health & Safety, France Titanium dioxide (TiO2) is the most commonly used white pigment in plastics and food packaging industry. TiO2 may be released from food packaging during storage, potentially entering the food chain and being ingested by consumers. Recent studies suggest possible toxic effects, including carcinogenicity 113 and reprotoxicity, however, a regulatory consensus is still lacking due to conflicting evidence. Since under current European Union (EU) legislation there is no specific migration limit (SML) for nanoparticles in food contact materials, assessing TiO2 migration from packaging into food is important for evaluating consumer safety. For this purpose, plastic containers from commercially-available dairy products and custom-made TiO2-spiked polypropylene materials were put in contact with 50% (v/v) ethanol and 3% (w/v) acetic acid, which were used here as food simulants. The migration assays were carried out under standard contact conditions of packaging use (as recommended by Commission Regulation N° 10/2011 for food contact migration testing), and under conditions of extreme mechanical degradation of the packaging. The TiO2 (nano)particles released in the food simulants were analysed by single particle inductively coupled plasma-tandem mass spectrometry (spICP-MS/MS) in mass-shift mode and using a high efficiency sample introduction system (APEX™ Ω) to avoid matrix effects from food simulants. For the dairy product containers and for the spiked polypropylene, results showed release of TiO2 particles of rather large sizes (average size: 164 and 175 nm, respectively) under mechanical degradation conditions, i.e when the polymeric structure is damaged. The highest amounts of TiO2 were observed in 50% ethanol after 10 days of storage at 50 °C (0.62 ng cm-2) for the dairy product containers and after 1 day of storage at 50 °C (0.68 ng cm-2) for the spiked polypropylene. However, the level of Ti released in particle form was very small compared to the total Ti content in the packaging and far below the acceptable migration limits set by European legislation. Release under standard contact conditions of use of the container was not measurable, thus the migration of TiO2 particles from this packaging to dairy products among storage is expected to be negligible. Trace-level determination of PFAS in water samples using LC-HRMS and a novel MOF sorbent in miniaturized SPE Vasilios Alampanos1, Zoi Terzopoulou2, Dimitrios Bikiaris1, Dimitra Lambropoulou1 1Aristotle university of Thessaloniki, Greece 2University of Ioannina, Greece A novel composite material comprising a modified iron-based metal–organic framework (MOF) dispersed in a polymeric monolith was developed and evaluated as a sorbent for miniaturized solidphase extraction (SPE).The material exhibits a highly porous architecture, enhanced specific surface area, and excellent chemical stability, offering improved selectivity and sorption capacity for perand polyfluoroalkyl substances (PFAS). Its tailored physicochemical properties make it a promising platform for sample preparation in complex environmental matrices. This innovation was integrated into a green analytical workflow aligned with the principles of Green Analytical Chemistry. The miniaturized SPE method was optimized using a Central Composite Design methodology by adjusting parameters such as sorbent amount, sample volume, microcolumn dimensions, and elution solvent, aiming for high recovery efficiency. Greenness was evaluated using the ComplexMoGAPI tool, confirming the method’s eco-friendly profile. The developed approach was applied to the determination of PFAS including PFPeA, L-PFBS, PFHxA, L-PFPeS, PFHpA, PFHxSK, L-PFHpS, PFOA, PFOSK, PFNA, L-PFNS, PFDA, L-PFDS, PFUdA, L-PFUdS, PFDoA, L-PFDoS, PFTrDA, and L-PFTrDS in various environmental water samples, including river water, marine water, urban runoff, and treated wastewater. Analytical determination was performed using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS). The method was validated for accuracy, linearity, sensitivity, repeatability, and reproducibility. Recoveries exceeded 70%, with relative standard deviations below 15%. This work demonstrates the potential of MOF-based sorbents for sustainable, efficient, and reliable environmental monitoring of persistent contaminants. 114 Funding: The research work was supported by the Hellenic Foundation for Research and Innovation (H.F.R.I.) under the “2nd Call for H.F.R.I. Research Projects to support Faculty Members & Researchers” (Project Number: 3616) CLUSTERING Consumer Perspectives on Food Fraud Vulnerability and Traceability: Insights from a Cross-Country Survey Matjaž Pirc Zveza Potrošnikov Slovenije, Slovenia Food fraud challenges consumer trust, public health and supply chain integrity, especially in globalized food systems. Addressing it increasingly relies on traceability and authentication tools, whose success depends on consumer acceptance. This study assessed consumer perceptions of food fraud and traceability across four EU countries, focusing on how supply chain, product, and institutional factors shape trust and willingness to pay—particularly for honey, a commonly fraud-affected product. A structured online survey was conducted in Italy, Germany, Portugal, and Slovenia (N≈550 per country), using nationally representative samples. It covered socio-demographics; purchasing behaviour, food fraud awareness and experience; trust in relevant actors, and a choice experiment (CE) on honey. The CE compared preferences for honey products varying in origin, authenticity testing tools, supply chain transparency, production method and price. Most respondents expressed concern for food fraud and a quarter had personally experienced it. While support for fraud prevention was strong, most saw it as the responsibility of authorities and industry, not consumers. Traceability ranked lowest among food purchasing criteria, yet origin and local production remained key trust signals. Trust was highest in short supply chains and certified labels, while digital authenticity verification tools (e.g., sensors or QR codes) raised skepticism. Over two-thirds found label information insufficient to assess authenticity and viewed packaged foods as more prone to fraud than fresh items. In the honey CE, consumers preferred products from their own country or the EU and favoured organic options. Tools verifying geographical origin were more accepted than those testing for adulteration. Willingness to pay for fraud prevention was low, but higher when technologies were part of familiar labels or schemes and did not increase price. Consumers expect industry and authorities to ensure authenticity. They trust origin labels and short supply chains but new traceability tools raise scepticism, especially if costly. Low willingness to pay for fraud prevention highlights a gap between consumer expectations and current approaches. Effective strategies should build on trusted cues and clearly show added value. Acknowledgments: Funded by the European Union. Views and opinions expressed are however those of the author(s) only and do not necessarily reflect those of the European Union or the European Research Executive Agency (REA). Neither the European Union nor the granting authority can be held responsible for them. This work was conducted as part of the Watson project, funded by the European Union’s Horizon Europe research and innovation programme, under grant agreement No. 101084265. 115 Galician mussel species identification via HRM analysis and ML for automated DNA Classification Lefkothea Karapetsi1, Kyriaki Psara2, Marilena Paraskeva2, Marios Kyriacou2, Macarena Garcia Silva3, Joaquin Garrido4, Panagiotis Madesis1 1Institute of Applied Biosciences-CERTH, Greece 2eBOS, Cyprus 3Kiwa España, S.L.U., Spain 4Consello Regulador da D.O.P. Mexillón de Galicia, Spain Mussel aquaculture is a significant economic activity in many coastal areas and especially for the region of Galicia in Spain. European legislation mandates the declaration of scientific and commercial identification on product labels. Seafood mislabeling is a widespread issue impacting authenticity, consumer choice, and potentially food safety due to undeclared allergens or species substitution. DNAbased authentication techniques are crucial for improving traceability, compliance, and transparency in the mussel supply chain by identifying mislabeling and substitution cases. The DNA-based authenticity kit within the THEROS toolbox offers a portable and rapid method for DNA extraction and automated analysis to verify species origin and product provenance, protecting consumers from adulteration. This kit provides full supply chain transparency and digital records for authenticity verification through standardized protocols, user-friendly software, and portable equipment. The methodology involves DNA extraction from product samples using robust, low-cost methods. Subsequent PCR and HRM analyses are performed on a portable device. PCR amplifies the COI gene region for species identification based on nucleotide differences. HRM analysis of amplified DNA sequences generates profiles and melting temperatures specific to each species and origin. For AI/ML DNA analysis, a dataset of 70 samples with DNA sequences from the COI gene and geographical origin (Chalastra, Greece; Galicia, Spain; Nea Michaniona, Greece) was used. Pre-processing included encoding target columns and shuffling data. K-mers representation (k=6) was applied to convert DNA sequences into numerical vectors using CountVectorizer, resulting in 2200 features for machine learning models. The dataset was split 75%- 25% into training (52 samples) and testing (18 samples) sets, ensuring representative class distribution. Multinomial Naive Bayes (MNB) and Multiclass Logistic Regression algorithms were used for DNA classification. MNB predicts class based on k-mer frequencies, while Multiclass Logistic Regression extends binary logistic regression for multiple classes. Evaluation metrics included Weighted Accuracy, Weighted Precision, Weighted Recall, and Weighted F1 Score to account for class imbalance. MNB results showed an overall accuracy of 94.4%, precision of 95.8%, recall of 94.4%, and F1 score of 94.7%, with only one misclassification for ‘Galicia, Spain’ samples. Multiclass Logistic Regression achieved 100?curacy, precision, recall, and F1 score, with no misclassifications across all three geographical classes, demonstrating its strong performance. The output is fed into a verification engine to ensure product authenticity and origin based on genetic signatures. THEROS project has received funding by the European Union under grant agreement No 101083579. 116 User-centred pilot testing of tools for organic and geographical indication of food products Maria Alejandra Rubio1, David Kocman1, THEROS Consortium2 1Jožef Stefan Institute, Slovenia 2THEROS project, Horizon Europe, Greece The EU-funded THEROS project aims to modernize the process of verifying organic and geographical indications of food products through the use of various technologies and innovations such as Earth Observation (EO) services, MEMS-based photonics systems, DNA authenticity kits, IoT sensors, blockchain-based traceability systems, and dynamic digital product passports. The project also aims to improve traceability, security, and transparency in the supply chain. To align these tools with the diverse needs of stakeholders—ranging from farmers and certification bodies to consumers—a user-centred design methodology was employed. This contribution presents a user-centred design of respective tools, starting with an analysis of the stakeholder landscape through co-design activities conducted in four THEROS pilots, resulting in various user scenarios. End-user needs were then consolidated into specific requirements and specifications for THEROS toolbox components. Personas representing key user groups were developed to guide the design and deployment of the THEROS tools in each pilot. The usability and perceived utility of each tool were assessed through structured user experience (UX) surveys targeting the primary stakeholders involved in the food value chain. These surveys evaluated factors such as ease of use, usefulness, efficiency, effort reduction, and visualization quality. Initial findings from the pilot sites indicate high user acceptance across multiple tools. In Serbia and Greece, EO tools and MEMS-based spectrometers can enable certification bodies and producers to monitor organic farming practices and detect adulteration in real-time. In Spain, DNA authenticity kits combined with blockchain traceability can improve the verification of mussel species and origin, building trust among retailers and consumers. In the Czech Republic, the digital marketplace and IoT network can enhance traceability and transparency in the delivery of regional products. Overall, the THEROS tools have been tested showing their potential to increase efficiency and trust in certification processes, improve decision-making through data visualization, and strengthen confidence in food authenticity. User feedback highlights the value of integrating these tools into a unified platform and emphasizes the importance of ongoing support and training to ensure continued engagement. To this end, this research underscores the value of participatory design and iterative evaluation in developing authenticity and traceability tools tailored to stakeholder needs. Acknowledgement: THEROS project (An integrated toolbox for improved verification and prevention of adulterations and non-compliances in organic and geographical indications food supply chain) has received funding under grant agreement No 101083579. It is funded by the European Union. 117 Biosafety assessment of raw materials and soil improvers for a safeuse in agriculture Lorenzo Nolfi1, Andrea Visca2, Victoria Crespo3, Patricia Oliveira3, Luciana Di Gregorio2, Elisa Clagnan4, Nicola Colonna2, Roberta Bernini1, Daniel Cantabella5, Ansa Palojärvi6, Avelino Álvarez Ordóñez3, Annamaria Bevivino2 1Department of Agriculture and Forest Sciences, University of Tuscia, Italy 2Department for Sustainability, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Italy 3Department of Food Hygiene and Technology and Institute of Food Science and Technology, Universidad de León, Spain 4University of Milano, Italy 5University of Vic - Central University of Catalonia, Futurlab - Can Baumann, Spain 6Natural Resources Institute Finland, Itäinen Pitkäkatu 4 a, Finland Recycling food processing residues streams using environmentally friendly technologies capable of inactivating harmful microbes represents a practical approach towards circular economy. These treated residues can serve as soil improvers for sustainable agriculture. The main aim of this study was to evaluate the biosafety of raw materials and soil improvers derived from sustainable, high-quality, and standardized valorisation processes designed to inactivate relevant biological hazards. Both culture-based and molecular analyses were employed to assess microbial pathogens and the presence of potential antibiotic resistance genes (ARGs) in raw materials and end products. Different DNA extraction methods, ranging from classical chemical methods to commercial kits, were used on different soil improver matrices (e.g. compost, biochar, digestate, biostimulant, and meat and bone meal) to standardize the experimental procedure. Preliminary screening of these DNA extraction methods enabled the identification of the most effective method for each type of sample, ensuring the retrieval of high-quality DNA suitable for subsequent metagenomic analyses. DNA extraction results were evaluated using Nanodrop, Qubit, and gel electrophoresis, leading to the development of a dedicated protocol for DNA extraction tailored to the different raw materials and soil improver matrices. Microbiological parameters were analysed in accordance with Standard Operating Procedures (SOPs) previously developed for the biosafety assessment of residues and soil improvers. Each sample was analyzed in quintuplicate, using 25 g for each replicate. Both compost and digestate met the established safety criteria, ensuring their suitability for sustainable agriculture. In conclusion, continuous monitoring of antibiotic-resistant bacteria (ARBs) and antibiotic resistance genes (ARGs) is essential to mitigate the spread of antimicrobial resistance. The integration of culture-based and molecular analysis will permit to evaluate the biosafety assessment of residues and soil improvers.