Published by ICC International Association for Cereal Science and Technology, Marxergasse 2, 1030 Vienna, on behalf of the MoniQA

A Network of Excellence (www.moniqa.org) funded by the European Commission within the Framework Programme 6 Vienna, Austria 2010

The content of the contributions are printed as received. No editorial changes have been made.

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ISBN 978-3-9501610-7-6 Printed in Austria

www.moniqa.org

Emerging and persisting food hazards: Analytical challenges and socio-economic impact
2nd International MoniQA Conference 8 10 June 2010 Qubus Hotel, Krakow, Poland

We would like to thank all participants at the Conference for helping make this event such a big success
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COMMITTEES
Scientific Committee: Roland Poms (Chair), ICC, AT John-Erik Haugen, Nofima, NO Wolfgang Kneifel, BOKU, AT Hans van Egmond, RIKILT, NL Martin Rose, Fera, UK Bert Ppping, Eurofins, DE Vasso Oreopoulou, NTUA, GR Marina Carcea, INRAN, IT Sue Paulin, ESR, NZ Michele Solfrizzo, CNR-ISPA, IT Clare Mills, IFR, UK Kim Anh To, HUT, VN Christoph van Holst, IRMM, BE Anton Alldrick, CamBRI, UK Mario Mazzocchi, Unibo, IT Miles Thomas, Fera, UK

Organising Committee: Marcella Gross (Chair), ICC, AT Anita Habershuber, ICC, AT Roland Poms, ICC, AT Daniel Spichtinger, RTDS, AT

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FOREWORD
The 2
nd

International MoniQA Conference: From Rome 2008 to Krakow 2010

Two years ago we came together in Rome for our first international MoniQA Conference on Increasing trust in rapid analysis for food safety which brought together a global audience of more than 200 food safety scientists, socio-economists, regulators, industry and trade representatives as well as media correspondents. For those of you who know MoniQA, it will come as no surprise that we have chosen Krakow, Polands second largest city and a leading centre of academic, cultural and nd artistic life, as the location for the 2 International MoniQA Conference. Not only is Polish cuisine, such as Pierogi, well known in Europe, Poland itself has emerged as a leading country in the group of nations that joined the European Union in 2004. From 1989 to the present day Poland has experienced many changes - political, economic and social. Food production systems have also changed and have been adapted to the legal requirements of the European Union. Today Poland has a population of 38 million, one of the most populous EU member states, with a sizable percentage of its workforce employed in agriculture which is why food and food safety play an important role in daily life. The theme of the 2 International MoniQA Conference, Emerging and persisting food hazards: Analytical challenges and socio-economic impact" forms a multidisciplinary bridge between the daily concerns of our citizens, economic considerations and food and nutrition science. Therefore it will not only bring together food quality and safety scientists and trainers but also food safety managers from industry, academia and regulatory bodies as well as representatives from food manufacturers, retailers and providers of rapid and high throughput analytical methods and instruments. Our conference is also of interest to food quality and HACCP practitioners, standardisation and validation organisations as well as representatives of consumer and trade organisations. In nine technical sessions the Second International MoniQA Conference will set out the state of the art concerning food safety standards, mycotoxins & phycotoxins, food allergens, chemical contaminants, microbiological contaminants, food additives, emerging technologies for food safety assessment and food authenticity. We will also include a poste r presentation and a MoniQA Best Poster Award. The conference includes results from the related projects BioCop (www.biocop.org) and CONffIDENCE (www.conffidence.eu). MoniQA (Monitoring and Quality Assurance in the Total Food Supply Chain) is a growing network and currently comprises some 145 institutions (core partners, advisory panel, and associated partners) from more than 35 countries across 6 continents. The MoniQA NoE (Network of Excellence) started 1 February 2007, it is funded by the European Union for a duration of 5 years. MoniQA aims at improving quality and safety of foods and feeds by setting new standards for method performance of rapid, alternative and modern high throughput tools to test foods and food products. Eight food quality and safety issues of top priority were identified by the international MoniQA expert panel that are being elaborated within the scope of the project by eight working groups. Each technical session of this conference is dedicated to a specific hot topic, and was organised by the corresponding working group. Since the last MoniQA conference in Rome our project has produced a variety of concrete results for the benefit of the food science community, a summary is given below. The 2 International MoniQA Conference will offer you a unique occasion to learn about recent developments in assuring food safety and quality world wide, in an attractive setting. Ample opportunity to meet old friends and to get acquainted to new colleagues in the field in a friendly environment; the poster and table top exhibition, and the gala dinner will also help to make a memorable experience. I look forward to meeting you in Krakow, and on behalf of the MoniQA consortium I would like to wish you an enjoyable stay and exciting three days of stimulating debates, discussions and networking. With best regards, Roland Ernest Poms Coordinator of MoniQA / ICC Secretary General
nd nd

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From Rome to Krakow: an Update of MoniQA Activities in Year Three In the third year of MoniQA, the network activities focused on strengthening integration through optimized sharing of information, expertise and potentially also infra-structure, to transform network achievements into tangible outputs for consortium and external stakeholder use integrated in MoniQAs sustainability plans, disseminating relevant information and training tools to stakeholders in the food safety and quality field. While several research collaboration agreements and new project proposals originated from the MoniQA consortium, the way for a sustainable legal entity of MoniQA was prepared. Major achievements in year 3 of MoniQA: Increased collaboration with related EU projects and initiatives by reaching formal agreements, organizing joint workshops, conferences and/or technical sessions, linking each others websites, teaming up for new research proposals and satellite research activities externally funded. Increasing content value and user-friendliness of the MoniQA partner/integration and infrastructure database. Opening public access of the MoniQA methods database, including feedback and market assessment (September 2009). Publishing the first harmonized guideline/protocol (AOAC/MoniQA): Guidance on a Harmonized Validation Protocol for Quantitative Food Allergen ELISA Methods (Journal of AOACI March/April 2010). A joint IUPAC/MoniQA protocol for validation of qualitative methods is in preparation. Submission of the ISO International Workshop Agreement on harmonizing bulk sampling procedures and resulting recommendation to ISO (chaired by AOCS and supported by MoniQA). Preparation of MoniQAs first Reference Materials Incurred for milk and egg allergen analysis. Preparation of 4 different MoniQA methods validation studies (inter-laboratory ring trials) for 1) quantitative milk and egg allergen ELISAs, 2) H2 and TH2 mycotoxins SPR-method developed by BioCop, 3) a multi-mycotoxins LC-MS-MS method, and 4) a microbiology dilution method. Design and testing of MoniQAs Tool Box for socio-economic impact assessment was extended to partners and interested parties at various DGs at the European Commission and FAO. The tool box should be ready-to-use by the end of 2011. Case studies to assess the socio-economic impact on various regulation changes and/or recent emergence of food hazards were elaborated in 2009. The launch of the peer-reviewed ICC/MoniQA Journal QAS Quality Assurance and Safety of Crops and Foods, published by Wiley Blackwell (March 2009). Numerous publications (research papers, reviews, fact sheets, etc.) in various scientific journals, including QAS, and through internet portals (especially the MoniQA website and linked partner sites and project sites. MoniQA FSTs Food Scientist Training courses enjoy increasing popularity. In year three a total of 10 MoniQA FSTs in 8 different countries hosted over 300 participants MoniQAs yearly training workshop for Codex Alimentarius delegates (CCMAS, in collaboration with the IAM Inter Agency Meeting) has become a lasting establishment. MoniQAs mobility programme awarded 70 travel bursaries and exchange grants in 2009 totaling 70,000.- . Preparations for the establishment of MoniQA2 legal entity within MoniQAs sustainability programme reached consensus of the principle concept of a non-for-profit association type and the business plan is under development. For offering MoniQAs outputs to the European and global food analytical community, MoniQA outputs were defined and designed as products to be marketed via MoniQA2 legal entity. Market assessment and acceptability testing of MoniQA offerings, in order to refine these products and to better respond to problems and gaps in the food manufacturing and food control areas, was done via surveys, feedback forms and meeting interactions. MoniQA partners were awarded 3 international research grants in 2009 (EC-FP7, OK-FSA, and bilateral Hungary-Turkey). Additionally, 4 joint research proposal were submitted to the EC-FP7 calls early 2010 and I proposal is in elaboration between EU and Canada. New cooperation agreements were reached within the framework of MoniQA between 6 partners (China-New Zealand, Hungary-Turkey, Austria-New Zealand). MoniQA contributed substantially to 4 Expert Summits on food safety and food security in 2009 (ASEAN-EU in Bangkok, Korean-EU in Brussels, Global ICC in Vienna, China-EU in Beijing). Information about MoniQA is now available in 14 national and regional languages.

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Table of Contents
COMMITTEES ...................................................................................................................... 2 FOREWORD ........................................................................................................................ 3 MONIQA PARTNERS........................................................................................................... 8 ADVISORY PANEL MEMBERS ......................................................................................... 13 ASSOCIATED PARTNERS ................................................................................................ 14 ORAL PRESENTATIONS................................................................................................... 16 OPENING SESSION........................................................................................................... 16 MONIQA WORKING TOWARDS SAFER FOODS THROUGH GLOBAL HARMONISATION OF FOOD SAFETY AND QUALITY MONITORING AND CONTROL STRATEGIES ............................................................................................. 16 ROLAND ERNEST POMS EU LEGISLATION FOR FOOD SAFETY IN THE FIELD OF CONTAMINANTS AND SOCIO-ECONOMIC CONSIDERATIONS ..... 17 FRANS VERSTRAETE SAFETY ASSESSMENT OF ENGINEERED NANOPARTICLES IN FOOD: THE IMPORTANCE OF DETECTION AND CHARACTERIZATION ........................................................................................................................ 19 KAREN TIEDE IMPLICATIONS OF RISK PERCEPTION FOR MANAGING AND COMMUNICATING FOOD RISKS TO THE PUBLIC....................................................................................................................................... 20 GENE ROWE SESSION 2 FOOD SAFETY STANDARDS/MANAGING FOOD SAFETY: STANDARDS, REGULATION AND THE SOCIETY ................................................................................... 21 FOOD SAFETY STANDARDS, INTERNATIONAL TRADE AND DEVELOPING COUNTRIES......................................... 21 SPENCER HENSON FOOD STANDARDS AND THEIR CONTRIBUTION TO FOOD SECURITY ............................................. 22 ELEONORA DUPOUY DIFFERENCES IN APPROACH TO APPLICATION OF REGULATIONS FOR DIOXINS (PCDD/FS) AND DIOXIN-LIKE POLYCHLORINATED BIPHENYLS (DL-PCBS) IN FOOD ................................................................................. 23 LIANA GIORGI, MARTIN ROSE RECENT DEVELOPMENT IN MULTI-CRITERIA EVALUATION OF REGULATION .................................................... 24 FABIO BARTOLINI SESSION 3 MYCOTOXINS & PHYCOTOXINS ............................................................... 25 DEVELOPMENTS IN THE EU ON MYCOTOXINS INCLUDING RISK ASSESSMENT, REGULATORY AND ANALYTICAL ISSUES 25 JOERG STROKA ISSUES CONCERNING FUSARIUM MYCOTOXINS IN THE 2007 & 2008 UK WHEAT HARVESTS .............................. 26 ANTON J. ALLDRICK

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MYCOTOXIN ISSUES IN TURKEY ......................................................................................................... 27 HAYRETTIN OZER TESTING A TOOLBOX FOR IMPACT ASSESSMENT OF FOOD SAFETY REGULATIONS: MAXIMUM LEVELS FOR T-2 AND HT2 TOXINS IN THE EU ........................................................................................................................ 28 MADDALENA RAGONA FUMONISINS: THE HIDDEN MENACE ........................................................................................... 30 GIANNI GALAVERNA SESSION 4 FOOD ALLERGENS .................................................................................... 31 CLINICAL THRESHOLDS AND THEIR APPLICATION IN RISK ASSESSMENT AND RISK MANAGEMENT .......................... 31 GEERT HOUBEN ALLERGEN MANAGEMENT: THE CORPORATE CHALLENGE. ........................................................................ 33 JAYNE HIPKISS EMERGING MULTI-SCREENING METHODS .............................................................................................. 34 BERT PPPING REFERENCE MATERIALS FOR FOOD ALLERGENS ANALYSIS .......................................................................... 35 PHILIPPE DELAHAUT RAPID LATERAL FLOW TESTS IN A HACCP-BASED APPROACH FOR ALLERGEN MONITORING........... 36 CHRISTINE GUTSCHELHOFER SESSION 5 CHEMICAL CONTAMINANTS ..................................................................... 37 ENVIRONMENTAL CONTAMINANTS AND CLIMATE CHANGE ........................................................ 37 BARBARA THOMSON THE IRISH DIOXIN CRISIS SIX DAYS THAT SHOOK THE NATION ................................................................... 38 WAYNE ANDERSON WHO ARE WE VALIDATING ANALYTICAL METHODS FOR: REGULATORS, ANALYSTS, OR THE CONSUMER? ............... 39 ROY MACARTHUR ACRYLAMIDE PHYSICAL REMOVAL FROM FOOD PRODUCTS ........................................................................ 40 MICHELE SUMAN SESSION 6 MICROBIOLOGICAL CONTAMINANTS ..................................................... 41 FOODBORNE OUTBREAKS: SURVEILLANCE, SOCIO-ECONOMIC IMPACT AND A PRECAUTIONARY TALE .......................................................................................................................................... 41 LISA OCONNOR IS FOOD A VECTOR FOR INTENTIONAL CONTAMINATION BIO-/AGROTERRORISM? ...................................... 42 BERNDT APPEL GENOME SEQUENCE COMPARISON OF CRONOBACTER SPECIES AND RELATED ORGANISMS ................................ 43 STEVE J. FORSYTHE DETECTION OF SALMONELLA IN FOOD - EXPERIENCES AND INNOVATIONS ................................... 44 HANNA-LEENA ALAKOMI SESSION 7 FOOD ADDITIVES ....................................................................................... 45 EMERGING AND PERSISTENT ISSUES WITH ARTIFICIAL FOOD COLOURS: METHODS OF ANALYSIS FOR NATURAL COLOUR ADDITIVES AS ALTERNATIVES TO ARTIFICIAL COLOURS IN FOOD AND DRINK .................................................... 45 MIKE SCOTTER

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COMPUTER VISION BASED IMAGE ANALYSIS FOR RAPID DETECTION OF ACRYLAMIDE IN HEATED FOODS ....................................................................................................................................... 46 VURAL GKMEN CHINA FOOD ADDITIVES HEALTH CODE(X) ........................................................................................... 47 ZHONGDONG LIU AN OVERVIEW OF METHODS FOR DETERMINATION OF TRANS-FATTY ACIDS IN FOOD .................. 48 MIECZYSLAW OBIEDZISKI SESSION 8 EMERGING TECHNOLOGIES FOR FOOD SAFETY ASSESSMENT (BIOCOP, CONFFIDENCE) ................................................................................................ 49 TARGETED AND UNTARGETED PROFILING FOR XENOBIOTICS ...................................................................... 49 GAUD PINEL REPLACEMENT OF THE MOUSE BIOASSAY FOR PHYCOTOXINS ..................................................................... 50 LUIS M BOTANA VETERINARY ANTIBIOTICS, MULTIPLEX DIPSTICKS, OPTICAL AND ELECTROCHEMICAL BIOSENSORS51 ANNE-CATHERINE HUET INNOVATIONS IN PESTICIDES ANALYSIS ................................................................................................ 52 JANA HAJSLOVA HEAVY METAL DETECTION USING CELL BASED AND SENSOR BASED ASSAYS .................................................... 53 DANILA MOSCONE SESSION 9 AUTHENTICITY AND TRACEABILITY ........................................................ 54 AUTHOR LOST WITHOUT TRACE: NEW ANALYTICAL APPROACHES TO TRACING THE ORIGIN OF FOOD .................................................................................................................................................. 54 PAUL BRERETON ISOTOPE ANALYSIS FOR THE CONTROL OF GEOGRAPHIC ORIGIN OF TYROLEAN MILK ......................................... 55 MICHA HORACEK RICE TRACEABILITY SYSTEM IN TAIWAN ............................................................................................... 56 SHIN LU IS IT ORGANIC? WHAT DO EXISTING FOOD AUTHENTICATION TECHNIQUES HAVE TO OFFER? .............................. 57 SIMON KELLY BIOLOGICAL BAR-CODE FOR THE DETERMINATION OF GEOGRAPHICAL ORIGIN OF FRUITS BY USING 28S RDNA FINGERPRINTING OF FUNGAL COMMUNITIES BY PCR-DGGE: AN APPLICATION TO SHEA TREE FRUITS .................. 58 ALY F. EL SHEIKHA POSTER PRESENTATIONS .............................................................................................. 59

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MONIQA PARTNERS
MoniQA Coordinator
International Association for Cereal Science and Technology (ICC)
ICC was founded in 1955 and has its headquarters and General Secretariat in Vienna, Austria. ICC is an independent, internationally recognized organisation of experts in the cereal and crops fields. It is a non-political, non-profit-making, neutral forum for cereal scientists and technologists. ICC is publisher of international standard methods and an important organizer of national and international events. Furthermore, ICC promotes international cooperation on a global, regional and national level and is a mediator between science and technology in research and practice. At present more than 50 countries are represented within ICC.

Ain Shams University (ASU)

Established in 1950, Ain shams University it is the third oldest university in Egypt. The university encompasses 15 faculties and 2 high institutes. It includes more than 180,000 students, 5,000 staff members, 4,000 assistant staff and more than 100 centres and special units.

Budapest University of Technology and Economics (BUTE)

Budapest University of Technology and Economics (BUTE) was founded in 1782. As a prestigious Hungarian higher education institute it is committed to differentiated, multilevel, high-standard education, founded on intensive basic training, research, development and innovation, and scientific qualification in technical and natural sciences and in certain fields of economic and social sciences. The Faculty of Chemical Technology and Biotechnology is one of the oldest faculties in the university. The Department of Applied Biotechnology and Food Science was established in 2007.

Campden BRI (CAMBRI)

Campden BRI is the world's largest, independent, membership-based organisation carrying out research and development for the food and drinks industry worldwide. It is committed to providing industry with the research, technical and advisory services needed to ensure product safety and quality, process efficiency and product and process innovation. In addition to its internal research programme, Campden BRI carries out contract R&D, analysis, consultancy and training on behalf of UK and other government departments, levy boards, industrial clients, the European Union and other international organisations.

Centre d'Economie Rurale (CER)

The Laboratoire dHormonologie (Laboratory of Hormonology) of the CER Groupe (Belgium) is mainly active in the agro-veterinary field. The Laboratory of Hormonology provides a large range of analyses for the detection of residues and contaminants. Taking advantage of large up-to-date animal facilities, the Laboratory of Hormonology provides pharmaceutical companies with the opportunity of performing pharmacokinetic, metabolic and residues studies for the development of new molecules.

Chinese Cereals and Oils Association (CCOA)

Founded in 1986, CCOA is a national scientific and technical organization for cereals and oils industry, a member of the Chinese Association for Science and Technology (CAST). CCOA has nine divisions: Grain Storage, Foods, Rice Products, Breads, Oils and Fats, Feed, Information and Automation, Grain Transportation, and Marketing, with 7,700 individual members and 1098 corporate members. CCOA has a good relation and cooperation with government departments, enterprises, research institutes, and universities.

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Consiglio Nazionale delle Ricerche, Institute of Sciences of Food Production (CNR-ISPA)

The main focus of the Institute of Sciences of Food Production (ISPA) is to develop research aimed at improving quality and safety of agri-food production, using technologies with minimal environmental impact. ISPA research is carried out using a multidisciplinary approach based on competences in various disciplines (chemistry, toxicology, microbiology, biotechnology, veterinary science, agronomy, biology and plant pathology) which are available within the five Research Centres.

Eurofins Analytik GmbH (EUROFINS)

Eurofins Analytik GmbH is one of Germanys leading commercial laboratories for chemical and biological analysis. Eurofins Analytik GmbH belongs to the Eurofins Scientific Group, created in 1987. With about US$ 600 million annual revenues and 7000 employees across 100 sites in 27 countries, Eurofins Scientific is a leading international group of laboratories providing an unparalleled range of testing and support services to the pharmaceutical, food, environmental and consumer products industries and to governments .

The Food and Environment Research Agency (FERA)

FERAs over arching purpose is to support and develop a sustainable food chain, a healthy natural environment, and to protect the global community from biological and chemical risks. Our role within that is to provide robust evidence, rigorous analysis and professional advice to Government, international organisations and the private sector. Climate change, food security and environmental sustainability are presenting the UK and indeed all other countries around the world, with significant, complex and often interrelated challenges. The Food and Environment Research Agency plays a vital role, increasingly on a world stage, in anticipating the issues, assessing the risks and gathering the evidence to guide policy response.

Fundacion Gaiker (GAIKER)

The GAIKER Foundation is a Technological Centre whose mission is to research and provide innovation and technology services to companies. Since its creation in 1985, GAIKER has contributed to the technological development and competitiveness of the business fabric through the generation, uptake, adaptation and subsequent transfer of knowledge to clients and founder members. Areas of expertise are in aparticular Plastics and Composites, Sustainability and Environment Technology, Recycling and Recovery, and Biotechnology.

Hacettepe University (HCTU)

Hacettepe University is a state university that ranks as one of the largest universities in Turkey with 34000 students enrolled in undergraduate and about 3500 students in postgraduate programs. The Department of Food Engineering at Hacettepe University was founded in 1977. The department aims to provide graduates with contemporary knowledge and skills that can be applied to design, develop and manufacture safe, high quality food products and production systems for the benefit of food industry and society.

Hanoi University of Technology (HUT)

HUT (Hanoi University of Technology) established in 1956, is a Government education institution at undergraduate and graduate levels. HUT has 1500 academic staff members, 54 of which are working in the field of Food Technology and related. The annual number of freshmen enrolled in the University is 3500 ( 150 in Food Technology).

Institut Pertanian Bogor (IPB)

The Department of Food Science and Technology (formerly Department of Food Technology and Human Nutrition) - Bogor Agricultural University is one of the oldest and largest departments of its kind in Indonesia. The department was founded in 1964 under the Faculty of Agricultural Technology and Engineering . The Department has a core competence in the area of food science and food technology, especially in the development of food chemistry, food microbiology, food process engineering, food analysis, food quality and safety.

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Institute of Environmental Science and Research (ESR)

ESR provides a range of scientific services to government organisations and commercial companies in New Zealand and the Asia-Pacific region. From applied science and research through to advanced scientific information systems, ESR provides specialist science solutions and commercial services tailored to its clients' needs.

Institute of Food Research (IFR)

The Institute of Food Research's vision is to be a world-leading contributor to harnessing food for health and controlling food-related disease. IFR is a not-for-profit company with charitable status. It is sponsored by the Biotechnology & Biological Sciences Research Council. IFR is the UKs only integrated basic science provider focused on food. O utcomes feed into national and international strategies, delivering advice and solutions for UK Government, public sector bodies, regulatory authorities, industry and consumers .

International Quality and Environment Services S.A. (Q-PLAN)

Q-Plan S.A. is active in the wider field of management consulting, advising the whole spectrum of enterprises and organisations. Moreover, Q-Plan is active in energy issues, with the main emphasis on the planning and the manufacture of renewable sources of energy.

The Interdisciplinary Centre for Comparative Research in the Social Sciences (ICCR)
ICCR was founded in 1986 as a private non-profit research organisation and is based in Vienna and Paris. The research centre specializes in strategic policy analysis using quantitative and qualitative methods in multi-disciplinary and comparative research. ICCR has four thematic research programmes and one overarching think-tank: (1) Transport Policy Analysis and Evaluation; (2) Social Policy Analysis Social Structures and Integration; (3) Environment and Sustainability; (4) Society, Technology and Research. (5) European Developments: Policies and Politics operates as a think-tank on the basis of the integration of research results from the four thematic programmes.

Joint Research Centre (JRC)

The Institute for Reference Materials and Measurements (IRMM) is one of the seven institutes of the Joint Research Centre (JRC), a Directorate-General of the European Commission (EC). It was founded in 1957 under the Treaties of Rome. Today IRMM is one of the world's leading reference material producers, expert adviser in food safety and quality and bioanalysis as well as a valued provider of reference measurement data.

National Food and Nutrition Institute (NFNI)

The National Food and Nutrition Institute was founded following the resolution of the Council of Ministers of April 12, 1963. It was established as the leading national scientific-research centre in the field of widely defined human nutrition and with contribution of the Food and Agriculture Organization of the United Nations in terms of advice and financing. The Institute employs now over 150 persons.

National Institute for Research on Food and Nutrition (INRAN)

INRAN, the National Institute of Research in Food and Nutrition, is a single governmentfunded research organization, based in Rome, Italy, which carries out basic and applied research in the various areas of study in food science and human nutrition. INRAN has a scientific and technical research staff of about 150 employees. The mission of the Institute is to perform independent, basic and strategic research into key issues of concern to industry, consumers and regulators and to spread knowledge by means of teaching activities at several levels.

National Technical University of Athens (NTUA)

The National Technical University of Athens (NTUA) is the oldest and most prestigious educational institution of Greece in the field of engineering and technology, and has contributed unceasingly to the country's scientific, technical and economic development since its foundation in 1836. NTUA is divided into nine academic Faculties (Schools), eight being for the engineering sciences, including architecture, and one for the general sciences.

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Nofima Norwegian Institute of Food, Fisheries and Aquaculture Research (NOFIMA)

Nofima is a business oriented research group working in research and development for the aquaculture, fisheries and food industry in Norway. Nofimas role is to provide research and solutions at an international level which will give a competitive edge throughout the value chain in cooperation with business actors and their professional organisations. Nofima was established on 1 January 2008, and has about 470 employees. The group's total turnover in 2008 was about NOK 470 million. The group's main office is located in Troms in northern Norway, while the research divisions are located in six places: s, Stavanger, Bergen, Sunndalsra, Avery and Troms.

Rheinische Friedrich-Wilhelms Universitt Bonn (UNI BONN)

The University of Bonn was founded almost 200 years ago and is considered to be one of Germany's and indeed Europe's most important institutes of higher education. As a home of learning to over 27,000 students, it enjoys an outstanding reputation both at home and abroad.

RIKILT - Institute of Food Safety

RIKILT is a Dutch independent scientific organisation. We carry out high-grade research into the safety, health and quality of Dutch food and feed and provide consultancy services to national and international governmental authorities. RIKILTs research activities are divided into different themes. Around 90% of the projects are covered by the themes: contaminants, veterinary drugs, animal feed, regulations, biological agents, food bioactives, authenticity & identity, and risk research. Together with a number of other institutes, RIKILT is part of Wageningen URs Agricultural Research Department (DLO). This department provides the framework within which Wageningen UR carries out legal research tasks for the government. The activities relevant to legislation and regulations are thereby separated from more market-oriented tasks and assignments, which are carried out within the holding company. RIKILT has some 200 employees.

RTD Services (RTDS)

RTDS is specialised in the development and management of research and technological (RTD) projects. We provide professional expertise in the organisation of RTD processes and the commercialisation of project results. RTDS serves universities and research institutes, industry and public authorities in all phases of their RTD projects.

Sichuan University (SCU)

Sichuan University, founded 1895, currently has the widest coverage of disciplines and the largest scale of operation in West China. The West China School of Public Health (WCSPH), also called Huaxi School of Public Health, has its origin in the curricular group of hygiene and public health which was set up in 1914.

Technical Research Center of Finland (VTT)

VTT is an impartial non-profit making expert organisation. Its objective is to develop new technologies, create new innovations and value added thus increasing customer's competitiveness. With its know how VTT produces research, development, testing and information services to public sector and companies as well as international organisations. VTT Technical Research Centre of Finland is the biggest contract research organisation in Northern Europe.

Tubitak Marmara Research Center (TUBITAK)

The Scientific and Technological Research Council of Turkey (TUBITAK) is the leading agency for management, funding and conduct of research in Turkey. It was established in 1963 with a mission to advance science and technology, conduct research and support Turkish researchers. TUBITAK Marmara Research Center (MRC), one of the research and development units of TUBITAK, was established in 1972 .

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Universit di Bologna (UNIBO)

The University of Bologna - probably the first University in the western world (its creation is dated 1088) is one of the most popular universities in Italy, with approximately 100.000 students. The Department of Statistics "Paolo Fortunati", established in 1982, develops a number of multidisciplinary research themes which include: statistical methodology, economic statistics, demographics, agri-food economics and statistics, and social and biosanitary statistics.

University of Food Technologies (UFT)

The University of Food Technologies was established in 1953 in Plovdiv, Bulgaria. UFT is the only academic institution in Bulgaria focused on all aspects of food science, engineering and technology. UFT educates an average of 3,300 students/academic year. The Department of Biotechnology was established in 1963 as part of the Technological Faculty at the University of Food Technologies.

University of Naples Federico II Department of Food Science (DSA)

The University of Naples, today called Federico II, was founded in 1224. It is currently the second largest university in Italy, after the University of Rome. It attracts more than 90000 students and employs a staff of more than 3000.

University of Natural Resources and Applied Life Sciences, Vienna (BOKU)

BOKU, founded 1872, nowadays is a "University of Life", with an emphasis on researching and teaching on sustainable use and protection of basic life resources. The excellence of BOKU is based on synergies between natural-, technical-, social-, life- and economic sciences which thematically focus on 6 fields of expertises, one of them "Food-NutritionHealth" with its assigned departments "Department of Food Sciences & Technology" and "Department for Agro-Biotechnology".

Vocal Tag Ltd (VTAG)

Vocal Tag is an Israeli start-up company focusing on the development of biotechnological electronic devices in the field of animal health and feed management. At present Vocal Tag is marketing its computerized rumination monitoring technology that has already been adopted by an Israeli management system manufacturer. The rumination sensor is integrated into a new generation of tags alongside other functions such as an activity sensor.

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ADVISORY PANEL MEMBERS

The MoniQA Advisory Panel provides scientific and strategic advice to the network. It includes individuals from a variety of backgrounds including researchers from related projects, end-users (policy-makers, planners, authorities) and other relevant opinion leaders. The following people are currently serving on the Advisory Panel: Joel Abecassis INRA-Institut National de la Recherche Agronomique, Unit de Technologie des Crales et des Agropolymres, Montpellier (FR) Elke Anklam Institute for Health and Consumer Protection, Ispra (IT) Marie-Noelle Bourquin International Standards Organisation, ISO TC 34 Foods Geneva (CH) Richard Cantrill American Oils Chemists Association St. Louis (USA) Selma Doyran Joint FAO/WHO Food Standards Programme Rome (IT) Samuel Godefroy Health Canada, Ottawa (Can) Margit Heinrich Deutsches Institut fr Normung e.V./ Department for Food Stuffs and agricultural products and office of CEN TC 27, Berlin (DE) David Lineback International Union of Food Science & Technology College Park (USA) Dominique Parent-Massin Professor of Food Toxicology, University of Bretagne Occidentale Breste, (France) Andr Pirlet (Chairperson) European Committee for Standardisation Brussels (BE) Nico Van Belzen The International Life Sciences Institute -Europe Brussels (BE) Jan Willem Van der Kamp TNO - Netherlands Organisation of Applied Scientific Research Delft (NL) Roger Wood Inter Agency Meeting Budapest (HU)

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ASSOCIATED PARTNERS
The following institutions have, by signing the Memorandum of Understanding, been approved as Associated Partners (Status May 2010):

AARINENA - Association of Agricultural Research Institutions in the Near East and North Africa (JO) AFBI - Agri-Food and Biosciences Institute (Northern Ireland) aha! - Schweizerisches Zentrum fr Allergie, Haut und Asthma (Switzerland) ARO - Agricultural Research Organisation, the Volcani Center (Israel) APCL - Ankara Provincial Control Laboratory (Turkey) ARC - Austrian Research Centers GmbH (Austria) BAC - BioAdvantage Consulting (France) HC - Bureau of Chemical Safety, Food Directorate, Health Canada (Canada) CGPRDI - China Grain Products R&D Institute ROC (Taiwan) CIRAD - Centre de Cooperation Internationale Recherche Agronomique pour le Dveloppement (FR) CLTAAS - Central Laboratory of Tianjin Academy of Agricultural Sciences (China) CRA - Istituto Sperimentale per la Cerealicoltura (Italy) Curtin University - Curtin University of Technology, Food Science and Technology Program (Australia) CUT - Cyprus University of Technology (Cyprus) DIL - Diagnostics Innovations Limited (United Kingdom) DM - Danismanik Muhendislik LLC (Denmark EGEFOOD - Ege University (Turkey) Elisa Systems - ELISA SYSTEMS Pty Ltd (Australia) ELISATEC - ELISA Technologies Inc (USA) EWOS - Ewos Innovation (Norway) FACTA - Food Allergen Control Training Analysis Pty Ltd (Australia) FIS Europe - Food Information Service Europe (Germany) Fonterra - Fonterra Cooperative Group Limited (Australia) FTC - Food Technology Centre (Egypt) FVMS - Faculty of Veterinary Medicine Skopje (Macedonia/FYROM) GCSL - General Chemical State Laboratory (Greece) IAS - Institute of Animal Science of LVA (Lithuania) ICETA/REQUIMTE - Instituto de Cincias Tecnologias Agrrias e Agro-Alimentares/Rede de Qumica e Tecnologia (Portugal) ICT - Institute of Chemical Technology, Prague (Czech Republic) IGV - Institut fr Getreideverarbeitung (Germany) INSA - National Institute of Health Dr Ricardo Jorge (Portugal) INTI - National Institute of Industrial Technology (Argentina) IQTC - Guangdong Inspection and Quarantine Technology Center (China) ITACYL - Instituto Tecnologico Agrario de Castilla y Leon (Spain)

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KRISS - Korea Research Institute of Standards and Science (Republic of Korea) KSL - Kalite Sistem Laboratories Group (Turkey) LATU - Technology Laboratory of Uruguay (Uruguay) lifeprint - Lifeprint GmbH (Germany) LVA - Lithuanian Veterinary Academy (Lithuania) MSM - Food Control Laboratories Inc. (Turkey) NAGREF - National Agricultural Research Foundation Cereal Institute of Thessaloniki (Greece) Neogen - Neogen Corporation (USA) NIB - National Institute of Biology (Slovenia) NINFS China CDC - National Institute of Nutrition and Food Safety, Chinese Centre for Disease Control and Prevention (China) PFTOS - Faculty of Food Technology (Croatia) Phylogene SA (France) ProteinLabs - ProteinLabs Inc. (USA) QUB - Queen's University Belfast (UK) R-Bio - R-Biopharm (Germany) RSSL - Reading Scientific Services Ltd (United Kingdom) RZZZ - National Public Health Institute (Republic of Macedonia) SAU - Sakarya University (Turkey) STCIQ - Shangdong Technical Center of Inspection & Quarantine (China) Suez Canal University - Suez Canal University Dpt. of Food Hygiene, Faculty of Vet. Med. (Egypt) TBS - Tepnel Biosystems (United Kingdom) UAC - University of Abomey-Calavi (Benin) UEA-ENV - School of Environmental Sciences, University of East Anglia (United Kingdom) UGhent - Ghent University (Belgium) UHOH - University of Hohenheim (Germany) UNFG - Universit di Foggia (Italy) Unilever (United Kingdom) UNIME - University of Messina (Italy) UNIMI - University of Milan (Italy) UNIPR - University of Parma (Italy) UNIROMA - Rome University La Sapienza (Italy) UNI-S - University of Stuttgart (Germany) UP - University of Pretoria (South Africa) UTP - University of Technology and Life Sciences (Poland) TU Wien - Vienna University of Technology (Austria) VITO NV (Belgium) VNIIZ - All-Russian Scientific Research Institute for Grain (Russian Federation)

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ORAL PRESENTATIONS Opening Session

MONIQA WORKING TOWARDS SAFER FOODS THROUGH GLOBAL HARMONISATION OF FOOD SAFETY AND QUALITY MONITORING AND CONTROL STRATEGIES Roland Ernest Poms ICC International Association for Cereal Science and Technology, Vienna, Austria, www.icc.or.at, and Coordinator of MoniQA www.moniqa.org

MoniQA (Monitoring and Quality Assurance in the total food supply chain) is an EU-funded Network of Excellence (NoE), which involves experts from around the globe collaborating to harmonise worldwide food quality and safety monitoring and control strategies. The initial network of over 155 scientists from 20 countries has grown to over 400 experts from over 35 countries from 5 continents in the first 18 months. MoniQA focuses on validation of and performance criteria/requirements for methods used to analyse foods and food products for safety and quality with the main focus being on rapid methods and their applicability and reliability in routine testing. The work involves validation guidelines, validation studies, design and development of reference materials/testing materials, and validation guidelines. MoniQA will play an important role in integrating European and worldwide food quality and safety research by creating a virtual laboratory for joint research, training, dissemination and mobility programmes. It will allow and actively promote sharing of data and knowledge, as well as of equipment, materials and personnel through creation of a global platform for food Q&S researchers. MoniQA will enable shared access to the worlds best research facilities, tec hnological platforms, databases, analytical tools and knowledge. MoniQAs harmonised food quality and safety control strategies will add value in the food chain and will improve consumer confidence worldwide. MoniQAs socio-economic impact assessment will enhance effectiveness and efficiency of new food quality and safety regulations within the EU and worldwide. Stay informed about what is going on in MoniQA: - Sign up to use the MoniQA Database for free at http://www.moniqa.org/methodsdb - Read our newsletter at www.moniqa.org/newsletter - Sign up to the ICC newsletter by sending an email to moniqa@moniqa.org

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Oral Presentations

EU LEGISLATION FOR FOOD SAFETY IN THE FIELD OF CONTAMINANTS AND SOCIO-ECONOMIC CONSIDERATIONS Frans Verstraete European Commission Directorate-General for Health and Consumers

General principles and objectives of food safety legislation The EU legislation on contaminants in food fulfils two essential objectives: the protection of public health and removal of internal barriers to trade within the EU. Following the principles and objectives of the General Food Law , food safety legislation shall pursue a high level of human health protection. To achieve this objective legislation shall be based upon risk analysis. Risk assessment shall be based on the available scientific evidence and undertaken in an independent, objective and transparent manner. Risk management shall take into account the results of risk assessment, other factors legitimate to the matter under consideration and the precautionary principle where appropriate. When international standards exist or their completion is imminent, they shall be taken into consideration in the development of any standard at EU level Legislation on contaminants in food Council Regulation (EEC) No 315/93 of 8 February 1993 laying down community procedures for 2 contaminants in food is the framework for the Community action on contaminants. This framework Regulation 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 -in order to protect public health, maximum levels for specific contaminants shall be established where necessary; -the consultation of a scientific body (EFSA) for all provisions which may have an effect upon public health is mandatory. Based on this framework Regulation, maximum levels for the following specific contaminants have been established by Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting 3 maximum levels for certain contaminants in foodstuffs : Challenges for EU legislation on contaminants in food To reduce the presence of contamination in the food supply, "prevention is better than cure". Therefore there it is important to encourage preventive actions. Prevention requires knowledge and acquiring knowledge requires research. The establishment of maximum levels is not contrary to prevention as maximum levels, established at a reasonably achievable level, stimulate food business operators to apply preventive actions all along the food chain in order to avoid the contamination of the food chain. Furthermore regulatory standards provide a benchmark against the effectiveness of the successful implementation of prevention programmes and provide a tool for control authorities to control the correct application of prevention measures by each actor in the chain.
1

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Legislation on contaminants needs continuously be updated to ensure a continuous high level of human protection and to address the challenges with which the risk managers are faced such as - emerging contaminants: Brominated flame retardants, (per)fluorinated compounds, Alternaria toxins - contamination incidents with "new" contaminants: melamine, mineral oil, - new risk assessments: non dioxin like PCBs, arsenic, - updated risk assessments: cadmium, PAH, mercury, ochratoxin A, lead, - developments in risk assessment approaches: * risk-benefit assessment : nitrates in vegetables * Margin of Exposure (MOE): genotoxic carcinogens such as aflatoxins, PAH, - changing production conditions /climate change: Fusarium-toxins, - international developments within the Codex Alimentarius: aflatoxins, The presentation The presentation will focus on the challenges the risk managers/regulators are faced with, how this leads to changes to EU-legislation, in full respect of the principles and objectives laid down in the General Food Law and the framework Regulation, and particular attention will be paid to the costbenefit considerations, the enforceability of the legislation and other socio-economic considerations.

Regulation (EC) 178/2002 of the European Parliament and of the Council of 28 January 2002 laying down the general principles and requirements of food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety (OJ l 31, 1.2.2002, p. 1) 2 OJ L37, 13.2.1993, p. 1 3 OJ L 364, 20.12.2006, p. 5

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Oral Presentations

SAFETY ASSESSMENT OF ENGINEERED NANOPARTICLES IN FOOD: THE IMPORTANCE OF DETECTION AND CHARACTERIZATION Karen Tiede karen.tiede@fera.gsi.gov.uk EcoChemistry Team, Fera, Sand Hutton, York YO41 1LZ, United Kingdom

Nanotechnology is a fast growing market and engineered nanoparticles (ENPs) are finding widespread applications including use in consumer products, medical applications, packaging materials, processing technologies and novel or functional foods. During their manufacture and use, human exposure and the releases of ENPs to the environment are inevitable (1). The proliferation of nanotechnology has therefore prompted concerns over the risks of adverse effects of ENPs on organisms in the environment and humans. However, research to address these concerns is still in the fledgling stages and the development and application of adequate analytical techniques for the detection and characterization of ENPs in food and natural samples is challenging (2). This presentation will give an overview of the challenges faced during engineered nanoparticle risk assessment. Current testing approaches and available analytical methods for the measurement, characterization, detection and chemical analysis of engineered nanoparticles in comlex matrices will be illustrated using real world examples and advantages and limitations of the approaches will be discussed. The implications for food safety and regulation and priorities for future research will be highlighted. Keywords: nanotechnology, nanoparticles References: (1) Tiede, K., Boxall, A.B.A., Tear, S.P., Lewis, J., David, H. and Hassellov, M. (2008) Detection and characterisation of engineered nanoparticles in food and the environment Food Additives and Contaminants 25(7):795-821 (2) Tiede, K., Hassellov, M., Breitbarth, E., Chaudhry, Q. and Boxall, A.B.A. (2009) Considerations for environmental fate and ecotoxicity testing to support environmental risk assessments for engineered nanoparticles Journal of Chromatography A 1216(3):503-509

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IMPLICATIONS OF RISK PERCEPTION FOR MANAGING AND COMMUNICATING FOOD RISKS TO THE PUBLIC Gene Rowe Wageningen University

Establishing the nature of food risks is only one part of the solution to the problem of protecting the public (consumers) from those risks. Alerting the public to the results of scientific research, and then persuading the public to behave appropriately, is a major additional part of the food safety problem. In this presentation I will outline some of the difficulties that arise in risk communication, and indicate their implications for food risk management. In particular, I will discuss how people perceive risks, and why peoples perceptions often appear perverse to scientists, risk managers, and others. The se psychological proclivities further affect how people respond to communications about food risk (and indeed, whether they even bother to attend to food risk information). Partly consequent on a number of high-profile food scares/crises (e.g. the BSE case), and their associated risk communications failures, risk managers, policy makers and regulators have sought to improve and broaden the concept and activities of risk communication so that it is more consultative, two-way and interactive. The concept of public engagement in policy making has thus become prominent in the thinking of organizations responsible for food safety, especially within Europe (e.g. in the UK, within its Food Standards Agency (FSA), and in Europe, within the European Food Safety Authority (EFSA)). Involving the public to a greater extent in food safety and risk management may provide a number of potential benefits (although few of these have been proven), but will result in many political and practical difficulties. Such problems are likely to be manifold in the case of emerging risks that require prompt response from responsible authorities as will be discussed here. Keywords: Risk perception, risk communication, risk management, food risks, public engagement

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Oral Presentations

Session 2 Food safety standards/managing food safety: standards, regulation and the society
FOOD SAFETY STANDARDS, INTERNATIONAL TRADE AND DEVELOPING COUNTRIES Spencer Henson University of Guelph, Canada & Institute of Development Studies, UK

In recent years, there has been growing concern about the impact of food safety measures on trade in agricultural and food products, and in particular on exports from developing countries. Of particular concern is the increasing importance of private standards, which have emerged as an important mode of market governance in the agri-food sector of many industrialised countries. This trend has raised profound questions about the role of public and private institutions in governing food safety, as well as food quality and the wider social and environmental impacts of the agri-food system. Embedded in this dialogue are three concerns about the impacts of private standards on developing countries. First, there are questions about the legitimacy of private modes of governance in areas that have historically been the preserve of public regulation. Second, there has been a heated debate about the rights and wrongs of private standards and especially the ways in which they influence the governance of agri-food value chains, exerting controls over production systems in developing countries, redefining quality and influencing the distribution of power and value between value chain participants. Third, there are accusations that private standards jeopardise the ability of developing countries to compete in export value chains governed by agri-food standards. There are widespread claims that developing countries are unable to meet the increasingly exacting food safety and other requirements of European markets and that they lead to the exclusion of smallholders from export value chains. This range of concerns highlights the importance of private food safety standards for global agri-food businesses, trade and development. Nevertheless, the heterogeneity of private standards makes analysis of their impacts difficult. Reflecting this diversity, there has been a lack of clarity about the ways in which private standards differ from public standards, the range of different actors involved in the creation, adoption and implementation of private standards and the extent to which public regulations drive the development of some types of private standards. This paper aims to provide some clarity to these debates.

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FOOD STANDARDS AND THEIR CONTRIBUTION TO FOOD SECURITY Eleonora Dupouy E. Boutrif, R. Clarke, M. Kenny Food and Agriculture Organization of the United Nations

Achieving food security for all is at the core of FAOs mandate which is pursued through the provision of technical assistance and policy advice to member countries. Food security is an evolving concept. In the 1950s-60s, food security was equated to self-sufficiency in major staples. Following the FAO World Food Conference of 1974, food security was defined as access to sufficient food. The current FAO definition of food security incorporates a number of additional considerations Condition when all people, at all times, have physical, social and economic access to sufficient, safe and nutritious food to meet their dietary needs and food preferences for an active and h ealthy life: Food safety is therefore now recognized as an intrinsic component of food security. According to WHO, every year there are about 4 billion cases of diarrhoeal diseases globally. . These illnesses exacerbate problems of malnutrition and reportedly account for 1.8 million deaths annually. Nowadays globally integrated production and distribution systems of animal feed and food products, and the greater volume and diversity of traded food, increase the vulnerability and challenges for food safety in both developed and less developed countries. Additional challenges for food safety arise from climate change and emerging foodborne pathogens. The rapidly changing context in which food is produced and marketed requires on-going vigilance, preparedness, international partnership and cooperation in order to anticipate and meet future food safety challenges. Effective food control systems focus on preventing food safety risks through fostering coordinated food safety management along the entire food chain. Codex standards, guidelines and recommendations establish the basis for food safety management internationally. Their pre-eminent position in food safety regulation is due to two main features: their content is rigorously science-based and they represent consensus among the 180 Codex member countries. Codex standards are explicitly recognized within the WTO SPS Agreement as the global reference for food safety in international trade. The paper deals with FAOs programme of work relating to the Organizations Strategic Objective D of Improved Safety and Quality of Food at all Stages of the Food Chain and explains the linkages with FAOs broader mandate to improve food security worldwide. The paper includes discussion of challenges for food safety management in Europe and Central Asia region. KEYWORDS: food security, food safety, Codex standards 1 FAO World Food Summit, 1996. The derived from the definition four pillars of food security are (i) availability, which includes food production and trade-originated food; (ii) stability of food supply, that is influenced by weather variability, food prices, political and economic factors; (iii) physical and economic access to adequate quantities and varieties of food and (iv) healthy food utilisation. 1 www.who.int 1 See the AGNDC website: www.fao.org/ag/agn/agns/index_en.asp

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Oral Presentations

DIFFERENCES IN APPROACH TO APPLICATION OF REGULATIONS FOR DIOXINS (PCDD/FS) AND DIOXIN-LIKE POLYCHLORINATED BIPHENYLS (DL-PCBS) IN FOOD Liana Giorgi , Martin Rose 1 and Annuradha Tandon 3 4 5 With support from Katharina Paul , Ron Hoogenboom and Rainer Malisch 1 ICCR Schottenfeldgasse 69/1, A-1070 VIENNA, Austria 2 The Food and Environment Research Agency, Sand Hutton, YORK YO41 1LZ. UK 3 current institutional affiliation Erasmus University Rotterdam, The Netherlands 4 RIKILT, Akkermaalsbos 2, 6708 WB Wageningen, The Netherlands 5 Community Reference Laboratory for Dioxins and PCBs in Food and Feed, CVUA Bissierstrasse 5, 79114 Freiburg Germany
1 2

Within the EU there exists a variety of legislation for dioxins and PCBs in food. This legislation covers not only limits but also methods for taking samples, methods of analysis, and details for monitoring the presence of these compounds in food and for official control of food. The limits include maximum limits and action limits, and a third tier of values referred to as target limits are also currently proposed. The regulations are set by DG SANCO and are enforced by National competent authorities supported by a network of the Community Reference Laboratory and National Reference Laboratories in each Member State. We explore the difference in approach to implementation of these regulations in different EU countries, and the advantages and disadvantages of this regulatory framework for dioxin and dioxinlike PCBs in food. The main argument is that the distinction within EU legislation between maximum and action levels is only tenable under specific institutional conditions regarding official controls and the monitoring of dioxin and dl-PCBs in food. Currently, there is large variation in the extent to which regular and systematic monitoring and official controls are carried out in EU countries. The risks entailed in this state of affairs are juxtaposed to the administrative costs implied by an upgrading of the European infrastructure.

Keywords:

dioxins, monitoring, official control, regulation

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RECENT DEVELOPMENT IN MULTI-CRITERIA EVALUATION OF REGULATION Fabio Bartolini D. Viaggi Department of Agricultural Economics and Engineering (DEIAgra), ALMA MATER STUDIORUM, Viale Fanin 50, 40127 Bologna, Italy.

Agricultural activity is affected by a very wide set of regulations and prescriptions due to agricultural and non agricultural policies. In the literature about agricultural policy, a growing attention has been paid to the issue of policy evaluation. The objective of this paper is to provide a literature review of the Multicriteria Analysis (MCA) applied to the context of policy and regulation evaluation. Such purpose implies to identify firstly the main issues of the policy evaluation process and then to identify the potential contribution of MCA as a decision support tool in a policy making context. In the literature, evaluation has been dealt through the measurement of the performance of a policy, program or regulation by way of effectiveness or efficiency concepts. In spite of the simple nature of these two criteria, the evaluation exercises are strongly affected by several problems, which are mainly due to the weakness in the definition and in the measurement of both policy objectives and policy impacts. Generally two main approaches have been adopted for policy evaluation purposes: a) Cost-Benefit Analysis, based on the monetisation of both costs and benefit, and; b) MCA, based on the measurement of a set of indicators and their aggregation taking into account the importance attached to each of them by the Decision Makers. MCA is a method that enables to provide a more robust evaluation with respect to Cost-Benefit Analysis mainly due to the expression of impact with physical and economic indicators rather than monetisation of the impacts. Such property, in addition to the flexibility of the method, allows to adapt the MCA to several contexts and to assess an evolution covering a very wide range of impacts (environmental, social and economic). Several MCA methods have been applied to agricultural and environmental regulation. Generally these methods differ for the adoption of different aggregation functions (single synthesising versus outranking methods), for the treatment of the uncertainty in the evaluation (eg. fuzzy versus non-fuzzy methods), and for the degree of DM involvement (participative methods, interactive methods, etc.). MCA has been invented as decision support in the choice of alternative projects when decision is effected by trade-offs between criteria and then has been extended to both ex-ante and ex-post policy analysis. However, the applications to policy evaluation still denote a wide room for improvement. In particular, a improved ability to support the decision process can be expected if the MCA is included in a consistent process of monitoring and data gathering, and both MCA and monitoring are developed in such a way as to provide a structured interaction with the DMs within a participative approach. In addition, a cautious integration between MC comparison techniques and policy analysis concepts (e.g. additionality) should be sought in order to avoid misinterpretations.

KEYWORDS. Multicriteria Analysis, Policy Evaluations, Decision Support, Indicators, Multiple impacts.

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Oral Presentations

Session 3 Mycotoxins & phycotoxins

DEVELOPMENTS IN THE EU ON MYCOTOXINS INCLUDING RISK ASSESSMENT, REGULATORY AND ANALYTICAL ISSUES Joerg Stroka Institute for Reference Materials and Measurements (IRMM), Retieseweg 111, B-2440 Geel, Belgium.

In the European Union the potential health risk due to possibly contaminated food and feed with mycotoxins is a seriously taken topic. As a result, any measures have to be sound, guarantee wholesome food (and feed) along the production chain from the farm to the fork (of the consumer) and taking into account the function of the common market in the EU. As a result, regulatory measures depend on a sound risk assessment which is based on sound analytical results, in order to impose (where needed) regulatory measures such as guidance limits, regulatory limits, temporary bans in addition to guidelines for the reduction of mycotoxins. This tasks involves various permanently established institutional bodies such as the European Commission (EC) itself, the European Food Safety Authority (EFSA), the European Committee for Standardization (CEN) in addition to the competent authorities of each Member State. This also includes all control laboratories and the National Reference Laboratories (NRL) together with the European Union Reference Laboratory (EU-RL), formerly known as Community Reference Laboratory (CRL). This laboratory network works throughout the whole measurement cascade applicable for official food and feed control in the EU and carries out common exercises with the aim to maintain a common high level measurement capacity. In addition to these official bodies, it must not be forgotten that various research projects funded by the EC are an important tool for the progress in this field. Over the last decades the EC has funded several Integrated Projects (IP) as well as Network of Excellence (NoE) to foster the scientific progress needed to support European policy. This presentation will give with specific examples an inside view on the current activities in the above mentioned field, focussing on analytical issues as sound analytical results are the basis for all further actions and the fact that that global harmonisation of food monitoring by harmonised analytical procedures is one of the pillars in the MoinQA NoE.

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ISSUES CONCERNING FUSARIUM MYCOTOXINS IN THE 2007 & 2008 UK WHEAT HARVESTS Anton J. Alldrick Campden BRI, Chipping Campden GL55 6LD United Kingdom

Within the European Union (EU) legislation exists, specifying limits for the presence of a number of mycotoxins in raw materials, ingredients and finished products, including those derived from cereals (Commission of the European Communities, 2006). This includes limits for deoxynivalenol (DON) and zearalenone (ZEA) mycotoxins produced by Fusarium spp. In terms of, Infection, growth of and mycotoxin production by Fusarial organisms a key determinant is climate (in particular rainfall at anthesis (flowering) in the cases of both DON and ZEA and, in the case of ZEA, prior to harvest). The United Kingdom is a Member State of the EU, producing in the region of 20-25 million tonnes of cereals per year (World Grain Council, 2007) and is, to a large degree, self-sufficient in terms of wheat. In common with other Member States, there has been a significant investment by relevant stakeholders not only to understand the biology of mycotoxin formation but also to institute appropriate management systems to ensure that consumer exposure is kept to a minimum. These include guidance to farmers and the development of risk assessment tools; surveillance programmes and customer-driven supplier quality assurance programmes. Historically, due to a combination of factors, it has proven to be relatively easy to ensure that UK-produced cereals meet limits set in EU legislation. However this status quo was challenged by climate-related events during 2007 and 2008. These concerned rainfall during the critical months of June (anthesis) and August (harvest). Consideration of UK Meteorological Office data (www.metoffice.gov.uk) indicates that precipitation in the UK during June 2007 was 136.2mm (187% of the 30-year average) and although slightly above average in June 2008 was again significantly higher than the 30-year average in August 2008 (135.8mm, 160% 30-year national average). In terms of contamination of cereals with either DON or ZEA, the consequences were that levels rose. In the case of one well established survey (Edwards, 2008), mean DON levels in UK wheat rose to 317 g kg-1 in 2007 and 670 g kg-1 in 2008 (compared to a previous 5-year mean of 217 g kg-1). Although mean ZEA levels remained static in 2007 (17 g kg-1) they rose to 127 g kg-1 (previous 5-year mean: 17 g kg-1). Inevitably, additional measures were instituted by the agri-food industry. These included modifications to mycotoxin risk assessment tools, use of rapid mycotoxin testing at intake and positive release programmes. Despite these measures, in spring 2009, it became evident that it was increasingly difficult for manufacturers to supply ingredients for certain foods (e.g. breakfast cereals) which met legal limits for ZEA. Following discussions between industry and the Commission as well as performance of appropriate risk assessments; a derogation was granted and regulatory limits temporarily raised (Commission of the European Communities, 2009). The events of 2007 and 2008 demonstrate that natural forces, outside of human control can have significant implications in terms of the occurrence of DON and ZEA within a staple food crop even in the developed world. They also vindicate the considerable investment made by stakeholders in the UK cereals chain to acquire a clear understanding of the occurrence of these mycotoxins and thereby develop appropriate management tools to ensure that both consumer safety and choice are maintained even when mycotoxin levels fluctuate abnormally. The author thanks the European Commission for financial support provided through the MoniQA Network of Excellence (FOOD-CT-2006-036337)) for preparation of this manuscript. Commission of the European Communities (2006) Commission Regulation (EC) No. 1880/2008, Official Journal of the European Union, L364, 5-24 Commission of the European Communities (2009) http://ec.europa.eu/food/committees/regulatory/scfcah/toxic/summary19062009_en.pdf Edwards, S (2008) http://www.hgca.com/document.aspx?fn=load&media_id=5202&publicationId=3498 International Grains Council (2007) World Grain Statistics 2006 KEYWORDS; CLIMATE-CHANGE, DEOXYNIVALENOL, WHEAT, ZEARALENONE

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MYCOTOXIN ISSUES IN TURKEY Hayrettin Ozer H. Imge Oktay TUBITAK MRC Food Institute, Turkey

Mycotoxins are toxic secondary metabolites produced by fungi. They are found in a wide range of food commodities around the world especially in countries with climates of high temperature and humidity which are favourable for mould growth and toxin production. Mycotoxin formation may begin in the field preharvest, continued or initiated postharvest and stored products. The most important mycotoxins in foods and feeds are aflatoxins, ochratoxin A, fumonisins, patulin, zearalenone and deoxynivalenol by means of incidence and human health consideration. Turkey is one of the most important food producing countries in the world. Geographically the regions of Turkey have subtropical climates with high ambient temperature and relative humidity providing optimal conditions for the growth of toxigenic fungi and consequently mycotoxin formation. Economical point of view, sometimes major agricultural export products of Turkey are faced with mycotoxin problem which leads to serious economical lost. In Turkey, major mycotoxin problems are aflatoxins contamination in hazelnuts, dried figs, pistachios and red pepper; ochratoxin A contamination in raisins and dried vine fruits could cause serious problems during exportation besides health consideration. Intensive research studies are carried out on risky commodities for occurrence, surveys, perevention and control of mycotoxins on local products of Turkey. Contamination of toxigenic fungi and mycotoxin formation are serious widespread problem in the world. On the worldwide basis, at least 99 countries had mycotoxin regulations or guidelines set for food and/or feed. Legal practices for mycotoxin levels in foods and feeds in Turkey are established by the Ministry of Agriculture Rural Affairs. In Europe consisting Turkeys main importer countries maximum levels of mycotoxins subject to the trade of agricultural commodities are regulated by European Union unless there are no limits determined by the Codex Alimentarius Comission.

Oral Presentations

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TESTING A TOOLBOX FOR IMPACT ASSESSMENT OF FOOD SAFETY REGULATIONS: MAXIMUM LEVELS FOR T-2 AND HT-2 TOXINS IN THE EU Maddalena Ragona* M. Mazzocchi*, A.J. Alldrick**, M. Solfrizzo***, and H.P. van Egmond**** * Department of Statistics, University of Bologna, Italy ** Campden BRI, UK *** Institute of Sciences of Food Production, National Research Council, Italy **** RIKILT - Institute of Food Safety, The Netherlands

In the EU, all major draft laws must be subject to a regulatory impact assessment. The aim of socioeconomic research in the MoniQA Network of Excellence is to develop a toolbox for impact assessment of food safety regulatory proposals. The aim of this contribution is to present a draft of such toolbox with its application to a case study, regarding a proposal on setting maximum levels of T2 and HT-2 toxins in cereals and cereal products. Such toolbox is based on multi-criteria analysis rather than cost-benefit analysis, which cannot be complete and reliable in most situations due to major difficulties: poor data availability; difficulty in isolating confounding factors like weather and market forces; probabilistic outcome of some actions; uncertainty in compliance levels; different timing in the occurrence and discounting of costs and benefits. We argue that a multi-criteria analysis method dealing with both fuzzy linguistic and stochastic quantitative variables - which take into account uncertainty - is the best way forward to an ex ante comparison of alternative policy options, especially in the food safety area, where the effect on public health - not precisely quantifiable - is the most important impact as giving the rationale for a regulatory proposal. The proposed toolbox involves a preliminary qualitative assessment of the likely impacts of each of the policy options considered, with a coding/scoring procedure, in order to identify the greatest impacts. Then, a feasibility filter considers availability of data necessary for impact quantification, in terms of monetary and time constraints for data collection. The subsequent quantitative assessment is performed with different methodologies for the most important impacts. Finally, a fuzzy multi-criteria analysis approach which allows for a combination of qualitative and quantitative measurements - is used to arrive to a ranking of the policy options. The alternative policy options considered for the case study are: 1) maintaining the status quo; 2) setting soft maximum levels; 3) setting strict maximum levels. Cereals susceptible to T2/HT2 contamination are wheat (durum and soft), oats, barley, maize, and rye, but the wheat and oats are the most susceptible. This type of regulation would prominently imply impacts on public health, conduct of businesses, public authorities, and SMEs. The application of multi-criteria analysis shows a dominance of the status quo option. All data used for this example are based on expert consultation and statistical databases, but, due to time constraints and the main aim of the research (testing the draft evaluation toolbox), the outcome of the analysis should not be taken as relevant for a real policy decision process. This case study shows the potential of the toolbox, which will be improved and tested with additional case studies in the remaining years of the MoniQA project. Keywords: regulatory impact assessment, food safety regulations, multi-criteria analysis, T-2 toxin, HT-2 toxin, cereals References Commission of the European Communities (2006). Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. Official Journal of the European Union, L 364, 5-24. Commission of the European Communities (2006). Commission Regulation (EC) No 401/2006 of 23 February 2006 laying down the methods of sampling and analysis for the official control of the levels of mycotoxins in foodstuffs. Official Journal of the European Union, L 70, 12-34.

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Directorate General Health and Consumer Protection of the European Commission Reports on tasks for scientific cooperation, report of experts (2003). Collection of occurrence data of Fusarium toxins in food and assessment of dietary intake by the population of EU member states. SCOOP Task 3.2.10. European Commission (2009a). Impact Assessment Guidelines. SEC(2009) 92. 15 January 2009. http://ec.europa.eu/governance/impact/commission_guidelines/docs/iag_2009_en.pdf European Commission (2009b). Part III: Annexes to Impact Assessment Guidelines. 15 January 2009. http://ec.europa.eu/governance/impact/commission_guidelines/docs/iag_2009_annex_en.pdf European Parliament and Council (2004). Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 on official controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules. Official Journal of the European Union, L 191, 1-52. JECFA (2001). 1016.T-2 and HT-2 Toxins. In Safety evaluation of certain mycotoxins in food WHO Food Additives Series 47 (http://www.inchem.org/documents/jecfa/jecmono/v47je01.htm) http://www.inchem.org/documents/jecfa/jecmono/v47je06.htm Munda, G. (1995). Multicriteria Evaluation in a Fuzzy Environment: Theory and Applications in Ecological Economics. Heidelberg: Physica-Verlag. Ragona, M. and Mazzocchi, M. (2008). Food safety regulation, economic impact assessment and quantitative methods. Innovation: The European Journal of Social Science Research, 21(2): 145-158. SCF (2001). Opinion on Fusarium toxins, part 5: T-2 toxin and HT-2 toxin. Adopted on 30 May 2001. http://ec.europa.eu/food/fs/sc/scf/out88_en.pdf SCF (2002). Opinion of the Scientific Committee on Food on Fusarium toxins. Part 6: Group evaluation of T-2 toxin, HT-2 toxin, nivalenol and deoxynivalenol. Adopted on 26 February 2002. http://ec.europa.eu/food/fs/sc/scf/out123_en.pdf SCOOP (2003). "Collection of occurrence data of Fusarium toxins in food and assessment of dietary intake by the population of EU Member States". http://ec.europa.eu/food/fs/scoop/task3210.pdf

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FUMONISINS: THE HIDDEN MENACE Gianni Galaverna, C. Dall'Asta, C. Falavigna, M. Mangia, S. Sforza, A. Dossena, R. Marchelli Department of Organic and Industrial Chemistry, University of Parma, I-43124, Parma, Italy

Fumonisins are a group of structurally related mycotoxins, produced mainly by Fusarium verticillioides and F. proliferatum, which are the most important seed-borne fungi associated with corn. Several reports on the fate of fumonisins during corn processing have shown that analytical methods often underestimate the levels of fumonisins in corn and derived food products because of low recoveries. We have recently demonstrated that different independent methods for the quantification of fumonisins B1, B2 and B3 in raw maize generally gave very different results: although all methods were validated and in spite of the use of common calibrators, a poor agreement about fumonisin contamination levels was obtained. These difficulties and these lack of consistency have been attributed to the occurrence of bound forms of these mycotoxins in foods. Indeed, this behaviour might be due to the binding of fumonisins to the food matrix or to the modification of their structure which lead to compounds not easily detectable by the normal methods of analysis. The presence of these hidden forms has been proved in several food products applying an alkaline hydrolysis protocol to release hydrolyzed fumonisins (HFBs) from the matrix. On the base of in vitro experiment with model compounds, it has been suggested that upon thermal treatment fumonisin B1 may react with starch and proteins to form covalent adducts. More recently, hidden fumonisins have been found also in low processed food (e.g. flour, bread, pasta) or raw maize suggesting the possibility of a physical interaction among the target mycotoxins and food macroconstituents such as proteins. Indeed, checking for the presence of these forms, significant amounts of bound fumonisins were detected in all the considered samples. The application of an in vitro digestion protocol to raw maize allowed for a very higher recovery in native fumonisins in comparison with the amount detected by routine analyses, suggesting that the interaction occurring among analytes and matrix macromolecules is associative rather than covalent. Depending on the analytical method as well as the maize sample, only 37% - 68% of the total fumonisin concentrations were found to be extractable from the analytical samples whereas upon digestion the amount of detectable fumonisins is almost double than the amount detectable with routine analytical methods. These findings have several implications on the accuracy of the currently used analytical methods, but also on the possible contribution of these hidden forms to the overall toxicity of a contaminated food or feed sample. Keywords: hidden fumonisins, masked mycotoxins, maize References C. DallAsta, G. Galaverna, M. Mangia, S. Sforza, A. Dossena, R. Marchelli. Free and bound fumonisins in gluten-free food products. Molecular Nutrition and Food Research, 2009, 53, 492-499. C. DallAsta, M. Mangia, F. Berthiller, A. Molinelli, M. Sulyok, R. Schuhmacher, R. Krska, G. Galaverna, A. Dossena, R. Marchelli. Difficulties in fumonisin determination: the issue of hidden fumonisins. Analytical and Bioanalytical Chemistry, 2009, 395:1335 1345.

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Session 4 Food allergens

CLINICAL THRESHOLDS AND THEIR APPLICATION IN RISK ASSESSMENT AND RISK MANAGEMENT Geert Houben M. Blom, M. Spanjersberg TNO Quality of Life, PO Box 360, NL-3700 AJ Zeist, The Netherlands Tel.++31 30 6944419; Fax. ++31 30 6944988; E-mail: geert.houben@tno.nl

In the EU, the use as ingredients in food of several major allergens or products derived thereof is to be labelled according to EU Directives 2003/89/EC and 2006/142/EC. This obligation only concerns the use of allergenic ingredients according to recipe. However, allergens my also be present in food due to cross contamination in food production facilities or contamination of raw materials or ingredients. Pele et al. (2007), VWA (2007) and Spanjersberg et al. (2010) have demonstrated the presence of considerable amounts of allergens in many food products that did not carry any warning for the presence of these allergens. The presence of allergens without accompanying warning obviously poses a risk to allergic consumers, as these individuals have no opportunity to judge the appropriateness of the concerning food products for them to eat (Spanjersberg et al. 2010; Sheth 2010). In contrast to the absence of any warning on many products that do contain certain allergens, there appear to be many food products in the market that carry a precautionary (often called may contain) labelling to warn consumers for the possibility of unintended presence of allergens. In many cases, such precautionary labelling seems not to be based on a relevant risk but is meant as a disclaimer in case the producer cannot exclude a risk for 100%. This seems to lead to a non-selective use of this precautionary labelling, which causes other problems, such as a reduced food choice for allergic consumers and devaluation of the information value of such warning (Health Council of the Netherlands 2007). The consequence of the dilemmas described above is that there are many products in the market with a warning while there is no or only a very small (negligible) risk and that, at the same time, there are many products without a warning that contain (sometimes very high amounts of) allergens. Precautionary labelling seems to provide the allergic consumers with no useful information anymore. Risk analysis principles can be applied to solve this problem and to bring guidance, harmonisation and transparency in information delivery regarding possible unintended presence of allergens in food products. For practical application of a risk analysis-based approach, a risk assessment methodology is essential. We developed a risk assessment method to quantify the number of allergic consumers that may suffer allergic reactions to specific levels allergens in food products or to calculate concentration action levels that can be based on predefined tolerable risks (Spanjersberg et al. 2007 and 2010, Kruizinga 2008). Clinical threshold data are of major importance in this methodology. Clinical threshold data are available for most major food allergens for which management of cross contamination risks is needed. Suitability for use in risk assessment will be discussed and it will be demonstrated how such clinical thresholds can be used in allergen risk assessment and risk management. Keywords: food allergy, thresholds, risk assessment, risk management, precautionary labelling, may contain labelling

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References: EU Directive 2003/89/EC of the European Parliament and of the Council of 10 November 2003, amending Directive 2000/13/EC as regards indication of the ingredients present in foodstuffs. Official Journal of the European Union 2003; L 308; 15-18. EU Directive 2006/142/EC amending Annex IIIa of Directive 2000/13/EC of the European Parliament and of the Council listing the ingredients which must under all circumstances appear on the labelling of foodstuffs. Official Journal of the European Union 2006; L 368; 110-111. Health Council of the Netherlands: Food Allergy. Report of the health Council of the Netherlands, publication number 2007/07, 2007, The Hague, The Netherlands. Kruizinga et al.: Probabilistic risk assessment model for allergens in food: sensitivity analysis of the minimum eliciting dose and food consumption. Food and Chemical Toxicology 2008; 46; 1437-1443 Pele et al.: Peanut and hazelnut traces in cookies and chocolates: relationship between analytical results and declaration of food allergens on product labels. Food Additives and Contaminants 2007; 24; 1334-1344. Sheth et al.: Role of food labels in accidental exposures in food-allergic individuals in Canada. Ann Allergy Asthma Immunol 2010; 104; 60-65. Spanjersberg et al.: Concentrations of undeclared allergens in food products can reach levels that are relevant for public health. Food Additives and Contaminants 2010; 27;169-174 Spanjersberg et al.: Risk assessment and food allergy: the probabilistic model applied to allergens. Food and Chemical Toxicology 2007; 45; 49-54 VWA. Fact sheet, Dutch Food and Consumer Product Safety Authority, July 2007.

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ALLERGEN MANAGEMENT: THE CORPORATE CHALLENGE. Jayne Hipkiss Mars Chocolate Europe

The presentation will define what is an allergen and the symptoms of an allergic reaction. The most common food allergens and their national prevalence will be stated. Rules for food labelling and advisory statements will be discussed. The presentation will conclude with a description of the allergen control plans utilized by the Mars food business to minimize allergen contamination. Key words: Allergen, allergen control plans

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EMERGING MULTI-SCREENING METHODS Bert Ppping Eurofins, UK

Food allergens are regulated in Europe, North America, Japan and several other countries to protect the allergenic consumer from inadvertent consumption of allergens. Labeling of products in Europe is based on allergens beings present as ingredients, and thereby are traceable via paper-trail. However, the actual risk of allergens comes from unintentional contamination of products, either by carry-over or by the inadvertent choice of the wrong, allergen-containing ingredient due to human error. In none of these cases, a paper trail would provide information that the final product contains allergens. This can only be done analytically. At present, allergens are typically detected using ELISA methods and sometimes PCR. However, both of those methods have their drawbacks. The most important one for ELISA is that the detection happens via an antibody which may also detect other, similar structures, possibly leading to false positives, or may not detect structures due to denaturation of certain proteins which could still be allergenic, possibly leading to false-negative results. In addition, only one allergen per ELISA can be detected, which makes ELISA testing economically challenging in food production sites dealing with multiple allergens. PCR on the other hand does not per se allow quantification of allergens and only detects the DNA of an allergenic compound. This could in some cases where protein but no DNA is present or where DNA but no protein is present also lead to false-negative and false-positive results. While both techniques are very useful in today's' routine testing, the ideal method would offer multiple screening abilities and direct detection of proteins (or peptides). Here, mass spectrometry offers a potential solution: it has the ability to screen for all 26 protein based allergens listed in annex IIIa of the European Allergen labeling directive and offers direct detection of the peptides derived from tryptic digestion. This presentation will present the work in this area of the MoniQA Allergen Working Group: the achievements and the obstacles to overcome.

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REFERENCE MATERIALS FOR FOOD ALLERGENS ANALYSIS Philippe Delahaut 1 2 3 3 4 5 6 V. Dumont , S. Kerbach , P. Johnson , C. Mills , B. Popping , S. Tmskzi , , R. Poms 1 CER Groupe, Dpartement Sant, Marloie, Belgium 2 Eurofins Analytik GmbH, Hamburg, Germany 3 Institute of Food Research (IFR), Norwich Research Park, Colney, Norwich, UK 4 Eurofins Scientific Group, Pocklington, Yorkshire, UK 5 Department of Applied Biotechnology and Food Science, BUTE, Budapest, Hungary 6 International Association for Cereal Science and Technology (ICC), Vienna, Austria
1

Food allergies constitute an increasing problem for several years. Currently, these allergies can not be cured. A complete elimination of the allergen in the diet is the only way to avoid an allergic reaction. Thus, the allergic consumers must know the exact composition of the food they eat. The European Directive 2007/68/EC sets out labelling rules in order to inform the allergic consumers of the presence of allergens in pre-packaged foodstuffs. However, these consumers are also concerned by the problem of cross-contaminations during the food processing, for which there is no legislation. The management of food allergens requires reliable analysis tools for the detection of allergens in food. Very few validation data are available for the comparison of allergen detection methods. This is certainly due to the lack of harmonized validation protocols and of recognized reference materials. Incurred reference materials are difficult and expensive to produce as each type and dose of allergen requires its own production run with particular care taken to ensure no cross-contamination occurs. However, the need in such recognized and confident materials is urgent. The MoniQA Working Group on Food Allergens will provide incurred reference materials with egg and milk proteins at various concentrations in two food matrices. Cookies were selected as the first foodstuff to be incurred. The preliminary tests have been carried out with the non-fat milk powder RM 1549 and with the spraydried whole egg for allergen detection RM 8445 from NIST (National Institute of Standards and Technology). Cookies were incurred with the two compounds at two concentrations before processing: 100 ppm or 1000 ppm. The dough (not baked cookies) and cookies were analyzed with casein, -lactoglobulin and egg kits from different providers. As expected, each kit has its own calibrators and its own quantification procedure. The second selected reference material was soy based infant formula. A pre ring trial was performed with 3 labs in order to assess if the two materials were suitable as reference materials for milk detection methods and if most of available ELISA test kits were able to detect the milk powder RM 1549 in the two matrices. The results have allowed contamination levels that will ideally be applied in the later validation study to be identified. In less than a year, two validated incurred reference materials with milk protein at various concentrations would be available for milk detection methods. Keywords: MoniQA, food allergens, Reference Material Incurred, ELISA kit, validation

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RAPID LATERAL FLOW TESTS IN A HACCP-BASED APPROACH FOR ALLERGEN MONITORING S. Haas-Lauterbach , M. Richter , K. Schmitt 1 R-Biopharm AG, An der neuen Bergstrasse 17, 64297 Darmstadt, Germany, info@r-biopharm.de 2 bioavid Diagnostics GmbH & Co. KG, Schlossgasse 17, 64807 Dieburg, Germany, info@bioavid.de Presented by: Christine Gutschelhofer
1 1 2

Allergen management in food companies is now becoming increasingly important. Under Article 5 of the EC Hygiene Code 852/2004, food companies are required to introduce, carry out and maintain regular hygiene controls based on HACCP principles. The Codex Alimentarius contains a general description of the HACCP principles. Thus, HACCP is defined as a system which is designed to identify the health risks from food and to implement preventive measures for their control. With sensitive people, even slight traces of allergens can trigger an allergic reaction which may result in anaphylactic shock in the severest cases, and this can be lethal if untreated. To ensure food safety for consumer protection, the products must be correctly labelled and cross contamination by contaminated raw materials, in production processes, during storage or transportation must be prevented. R-Biopharm supports food companies when optimising the allergen management system by offering rapid allergen tests for analysing processes. Food manufacturers should use fast and reliable processes to check for allergen contamination in food production. The increased importance of these detection methods is indicated by the current debate and the fact that some of these methods have been officially validated and approved. Immunochemical test methods have become established due to the ease of preparation and their increased efficiency. Over and about the biochemical detection reaction is specific and determinations can also be carried out in complex matrices. Lateral flow tests are qualitative immunochemical methods using test strips. Because they are fast and sensitive, lateral flow tests can be applied for allergen screening and hygiene controls in food companies. They are an inexpensive way to ensure that food production is allergen-free. R-Biopharm offers a series of lateral flow tests for differentiating between various food allergens such as milk, egg, peanuts, mustard, almonds, hazelnuts and sesame. Other rapid tests for the detection of allergens such as soy, wheat and celery will follow. Allergen residues can be detected in surface samples or rinsing waters (CIP), food end products and raw materials. The tests are quick to carry out, taking approximately 15 minutes without laboratory equipment. The extracted sample is pipetted into a reaction vessel which contains the specific antibodies ready for use. If the sample contains the antigen in question, an antigen-antibody complex is formed in the reaction vessel which is detected on the test strips. The qualitative evaluation is carried out visually. The presence of two coloured lines on the test strip, the control line and the test line, indicates a positive result whereas the presence of one coloured line indicates a negative result. The sensitivity of the specific lateral flow tests is comparable to that of commercial ELISAs for the matrices under investigation. Health risks due to contaminated food can be detected and prevented in time.

Keywords: Allergen Management, Hygiene Control, HACCP, Lateral Flow Technology, Immunochemical Analytical Methods

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Session 5 Chemical contaminants

ENVIRONMENTAL CONTAMINANTS AND CLIMATE CHANGE Barbara Thomson Institute of Environmental Science & Research Ltd, Christchurch, New Zealand

In a food safety context, environmental contaminants including dioxins, polycyclic biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs) and heavy metals are groups of unwanted chemicals, ubiquitous in the environment, that may occur in food products. Climate change, defined as a change in the distribution of weather over periods of time, may be due to natural processes such as solar changes and volcanic eruptions, and/or persistent human activities like driving cars, farming and burning coal. Some evidence suggests the climate is becoming hotter, on average, and more variable, resulting in rising sea levels, warmer oceans and more extreme events such as floods, storms, cyclones, droughts and landslips. The aim of this work was to consider environmental contaminants in foods and feeds in the light of climate change. Dioxins and PAHs are both groups of environmental contaminants that are released into the environment, and thence uptake into the foodchain, as a result of combustion that may be natural, such as forest fires and volcanoes, or due to human activities including power generation, waste incineration and car exhausts. In addition, dioxins may be formed as unwanted bi-products of manufacturing processes, notably the defoliant Agent Orange. Although manufacture was banned in 1979, PCBs may enter the foodchain because of their ongoing release into the environment from poorly maintained hazardous waste sites; illegal or improper dumping of wastes and leaks or releases from electrical transformers containing PCBs; PCBs may also be released into the environment by the burning of some wastes in municipal and industrial incinerators. Arsenic, cadmium and mercury occur naturally in the earths crust and may enter the foodchain following release into the environment from volcanic eruptions, forest fires and/or industrial activity such as coal fired power plants, mining and smelters, use in thermometers, batteries, stabilisers and pigments for plastics, ceramics and enamels, components of alloys, electrical switches, agricultural practices, wood preservation, metallic platings and fuel additives. With respect to climate change, increased flood events may result in mobilisation of contaminated agricultural soils and industrial sites as evidenced in Central Europe in 2002 and following Hurricane Katrina. Warmer oceans facilitate the methylation of mercury and the subsequent uptake of methyl mercury in fish and mammals, thus increasing human dietary exposure to toxic methyl mercury. Evidence supports that climate change is altering natural biological systems, and increasing the uptake of heavy metals into the foodchain. Increased forest fires are resulting in increased release of environmental contaminants. On the positive side, climate change policies, to reduce carbon emissions, are driving energy efficiencies, leading to reduced emissions of dioxins, PAH and heavy metals.

Keywords: environmental contaminants, dioxins, PAHs, PCBs, heavy metals, climate change

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THE IRISH DIOXIN CRISIS SIX DAYS THAT SHOOK THE NATION Wayne Anderson Food Safety Authority of Ireland

In December 2008, the Irish Authorities ordered the largest recall of food products ever seen in the State following the discovery of dioxin contamination of pork and beef products. Close collaboration between the authorities in Ireland and colleagues in other Member States allowed the rapid identification of a common source of contamination, this being feed produced in one plant from recycled bread manufactured using a direct heating process. The original source of the contamination is thought to be recycled transformer oil used in the direct drying process. The rapid identification of the source of contamination in turn meant contaminated meat and meat product could be removed from sale very quickly and thus consumer protection was ensured, both in Ireland and in other countries. Whilst only approximately 8% of the national pig herd was exposed to contaminated feed, the accepted level of traceability in the pork processing industry, which complies with the minimum legal requirements, meant that it was not possible to distinguish between contaminated and noncontaminated pork in production representing 98% of Irish pork entering the food chain. Therefore a decision was taken to recall all Irish pork and pork products produced from 1 September 2008 up to the date of the recall in December. By way of contrast, only 0.02% of the national beef herd was exposed to contaminated feed and traceability in the beef sector meant that all of this product could be traced and withdrawn from the food chain. Risk assessments carried out by FSAI and EFSA, including an estimation of the likely impact on human dioxin body burden arising from consumption of potentially contaminated meat, confirmed that as a result of the swift action taken to recall products, there had been no unacceptable risk to consumers health arising from the contamination incident. Key words: dioxin, pork, crisis, Ireland

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WHO ARE WE VALIDATING ANALYTICAL METHODS FOR: REGULATORS, ANALYSTS, OR THE CONSUMER? Roy Macarthur 1 2 3 4 Martin Rose , , Bert Ppping , Franz Ulberth , Roland Poms 1 FERA, UK, 2 Eurofins, Germany, 3 JRC-IRMM Belgium, 4 ICC Austria
1

MoniQA ("Monitoring and Quality Assurance in the Food Supply Chain) is a Network of Excellence funded by the European Union. This Network of Excellence aims to make food safer by harmonising methods for food analyses. Part of this process involves the development of new methods, analytical criteria and standard methods to be used for food control and the clear communication of the capabilities of analytical methods. New technologies and analytical research enable us to measure new and emerging food contaminants that may be a threat to health of the consumer and ingredients that may raise issues about the authenticity of a product. Once risk assessment confirms the threat, legislation may be enforced to limit the amount of these substances present in food that is sold. To ensure that analytical methods used to monitor food are sufficiently robust and appropriate for use, regulations on maximum limits can be accompanied by analytical criteria that such methods must meet before they can be used for enforcement. There is also a demand by food control laboratories both for standard methods that can be readily established within a laboratory and which are known to meet the required criteria and for simple procedures for demonstrating that methods meet the criteria. We present a brief summary of the general approaches to using criteria acceptable performance of analytical methods, and demonstrate how the use of a simple graphical representation of measurement uncertainty can be used as a tool for describing the performance of methods that is suitable for analysts, enforcement authorities, policy makers, producers and consumers. Key words: validation, standard methods, analytical criteria, uncertainty, food control

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ACRYLAMIDE PHYSICAL REMOVAL FROM FOOD PRODUCTS Michele Suman b b M. Anese , M. C. Nicoli a) Barilla SpA Food Science & Research Labs, Via Mantova 166, 43100 Parma (Italy) m.suman@barilla.it b) Dipartimento di Scienze degli Alimenti, University of Udine, Via Sondrio 2/A, 33100 Udine, Italy monica.anese@uniud.it; mariacristina.nicoli@uniud.it
a

The most important acrylamide dietary sources include potato chips, french fries, roasted coffee and bakery products such as bread, crisp bread, biscuits, crackers, breakfast cereals: due to the great consumption among people of different ages and in different countries, worldwide efforts have contributed to identify potential routes to reduce acrylamide levels in foods and consequently consumer exposure. These are relevant to mitigations strategies, which include agronomical interventions (i.e. selection of raw materials with low sugar and asparagine contents), and technological strategies (e.g. chemical and biotechnological pre-treatments, thermal input and moisture control, formulation changes). The physical removal of acrylamide from the nished product can be regarded as an innovative approach, which has not been fully investigated. With respect to the other mitigation strategies, acrylamide removal is conceptually different. In fact, while the former are aimed at lowering possible acrylamide formation during heating, the objective of acrylamide removal is to physically remove the molecule after the heat process has been completed. The aim of the present study was to investigate the possibility to physically remove acrylamide from nished products (i.e. biscuits and potato chips), by considering different combinations of pressure, temperature and time. The percentage of acrylamide removal from the biscuits and potato chips increased with the increasing of water activity, suggesting that water plays an important role (water-food interactions, water stripping effects) inuencing the process. Acrylamide removal was higher from the biscuits than from the potato chips, being 43% and 18% the maximum percentage of acrylamide removed from the bakery product and the potato derivative, respectively. Such a difference could be attributed to a matrix effect, e.g. interactions between acrylamide and other food components, and/or to a hurdle effect exerted by the super cial lipid lm of the fried potatoes against acrylamide removal. A further consideration is that acrylamide removal is higher at shorter process times: it can be hypothesized that the vacuum treatment of an acrylamide-containing food, by favoring carbon dioxide removal, may promote the formation of the decarboxylated Schiff base, which subsequently reacts to form acrylamide. With respect to these preliminary results, more in-depth studies are needed to evaluate the impact of the technology on the sensory properties and to understand the role of each variable for maximizing acrylamide removal, while minimizing its contemporaneous formation by means of the vacuum process. Keywords: Acrylamide, Food, Physical Removal REFERENCES Friedman, M.; Levin, C. E. (2008), Journal of Agricultural and Food Chemistry, 56, 611-614. Zhaoyang, L. (2003), Patent No US2003/0219518 A1. Nicoli, M. C.; Anese, M. (2006), Patent No PD2006A000332. Taeymans, D.; Wood, J.; Ashby, P.; Blank, I.; Studer; A., Stadler, R. H. (2004), Critical Reviews in Food Science and Nutrition, 44, 323347. Anese, M.; Suman, M.; Nicoli, M.C. (2010), Food Chemistry, 119, 791-794.

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Session 6 Microbiological contaminants

FOODBORNE OUTBREAKS: SURVEILLANCE, SOCIO-ECONOMIC IMPACT AND A PRECAUTIONARY TALE Lisa OConnor Food Safety Authority of Ireland, Abbey Court, Lower Abbey St., Dublin 1, Ireland.

The primary aim of surveillance for foodborne outbreaks is the prompt identification of any unusual clusters of infectious intestinal disease (IID), potentially transmitted through food, which might require a public health investigation or response to prevent further cases. IIDs may also be transmitted by person-to-person spread or through contact with animals and they may be waterborne or airborne. Sometimes more than one mode of transmission may be involved in an outbreak. The European Food Safety Authority (EFSA) reported a total of 5,332 foodborne outbreaks notified across the 27 member states, during 2008. This involved 45,622 human cases, 6,230 hospitalisations and 32 deaths. Salmonella species were responsible for the largest number of outbreaks (35.4% of all outbreaks), followed by viruses (13.1%), bacterial toxins (9.8%) and Campylobacter species (9.2%). The most important food vehicles identified were eggs and egg products (23.1%), pig meat and products thereof (10.2%) and mixed or buffet meals (9.2%). Eggs and egg products, and bakery products were mostly associated with Salmonella Enteritidis outbreaks, whereas pig meat was linked to Trichinella spp. and Salmonella spp. outbreaks. The viral outbreaks were mainly caused by crustaceans, shellfish and molluscs followed by mixed or buffet meals. Every illness has a socio-economic impact. In the case of IIDs some researches have analysed the impact of a single pathogen or a specific outbreak, whereas others have estimated the impact of all IIDs in a country. Typically these studies focus on medical costs, the cost of loss of productivity and of premature death. Foodborne outbreaks also result in costs to the implicated food business (and possibly by association to other businesses producing similar food), the regulatory authorities and the public health departments involved in investigating the outbreak. During 2008, the Food Safety Authority of Ireland had first hand experience of the socio-economic impact of an international foodborne outbreak. In July of that year an outbreak of Salmonella Agona was identified through laboratory based surveillance. A temporal cluster of 6 cases was identified in Ireland and an increase in S. Agona cases had been noticed in the United Kingdom (UK). A total of 163 cases were identified as being part of the outbreak from 10 European countries, with the majority of the cases in the UK. The outbreak was linked to an Irish company producing cooked meats which were sold for use as ingredients in a range of chilled and frozen products. In total 47 products were affected. The investigation and control of the outbreak was made difficult by the complex nature of the distribution chain. Due to the intermittent contamination of the cooked meats in this outbreak, laboratory based surveillance (involving phenotypic and molecular typing) proved vital in the early detection of the outbreak and identification of its source. Keywords: foodborne outbreaks, infectious intestinal disease, surveillance, socio-economic impact
Buzby JC and Roberts T (2009) The Economics of Enteric Infections: Human Foodborne Disease Costs. Gastroenterology 136:1851-1862 European Food Safety Authority (2010) The Community Summary Report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in the European Union in 2008. EFSA Journal 1496 [288 pp.

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IS FOOD A VECTOR FOR INTENTIONAL CONTAMINATION BIO-/AGROTERRORISM? Berndt Appel A. Hensel Federal Institute for Risk Assessment, Dept. of Biological Safety bernd.appel@bfr.bund.de

The food chain is considered as a possible target for bioterrorist attacks. The internationally published quantitative microbial risk assessments (QMRA) are characterized by a great heterogeneity in all aspects related to the analysiss (aims, data, process descriptions, modeling techniques and software). Integral part our concept of risk assessment is the development of a knowledge base on food matrices, food production processes, process parameters, microbial agents and predictive models. An EU list of biological agents (and toxins) of security concern is under development. The criteria for an agent (or toxin) to pose a threat for food chains and therefore for humans have to be evaluated. Using the example of the food chain cattle data will be collected about biological agents (growth, inactivation, tenacity etc), analytical data (sample preparation, detection systems and detection limits) and food data (ingredients, processing, consumption). Concerning food chains all characteristics inside and outside food processing have to be evaluated (critical control points, vulnerability to unexpected threats, provider/distributors, amounts...). To identify critical vulnerability points, scenarios will be modelled and analyzed. Activity recommendations can be derived from these scenarios based on risk assessment. In case of intentional contaminations of the food chain our data will be helpful for the risk management to overcome a bottleneck in supply and to manage disposal and/or treatment of contaminated food and/or contaminated production plants. The major goal of our research project is the development of an information- and database platform for governmental and industrial partners in favor of having admission to the same pool of data for risk assessment and management measures. The database structure integrates all sources of information (agents, detection methods, supply chain data, process chains, ingredients, analyzed scenarios and management responses), and will be accessible via internet for approved partners and can in part also be handled as local installation, The software allows chain structure analysis, assessment of risks, cost-benefit analysis, VACCP analysis and so on, and gives the possibility to integrate early identification and warning systems. Depending on the available and integrated (experimental) data, the developed software and data infrastructure can become an integral part of a strategy to safeguard the food (and feed) supply chain in case of intentional contaminations and thus will be a valid counteraction against bio-/agroterrorism. Literature: R. L. Buchanan and B. Appel (2010), International J. of. Food Microbiology: Combining analysis tools and mathematical modeling to enhance and harmonize food safety and food defense regulatory requirements

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GENOME SEQUENCE COMPARISON OF CRONOBACTER SPECIES AND RELATED ORGANISMS Steve J. Forsythe 1 1 2 E. Kucerova , S. Joseph , M. McCelland 1 School of Science and Technology, Nottingham Trent University, Clifton Lane, Nottingham, UK, NG11 8NS. 2 Vaccine Research Institute of San Diego, San Diego, California, USA.
1

Members of the Cronobacter genus (formerly Enterobacter sakazakii) have become associated with neonatal infections and in particular contaminated reconstituted infant formula. However this is only one perspective of the organism since the majority of infections are in the adult population, and the organism has been isolated from the enteral feeding tubes of neonates on non-formula diets. In recent years methods of detection from food and environmental sources have improved, though accurate identification has been problematic. The need for robust identification is essential in order to implement recent Codex Alimentarius Commission (2008) and related microbiological criteria for powdered infant formula (intended target age 0-6 months). Genomic analysis of emergent pathogens is of considerable advantage in both improving detection methods, and understanding the evolution of virulence. One ecosystem for Cronobacter is on plant material which may enable the organism to resist desiccation, adhere to surfaces, and resist some antimicrobial agents. These traits may also confer survival mechanisms of relevance in food manufacturing and also virulence mechanisms. Complete genome sequencing and mulitlocus sequence typing (MLST) of Cronobacter strains has enabled a more detailed understanding of the organism including the identification of putative virulence genes, location of desiccation resistance genes, and the diversity of strains within the genus. Subsequently comparisons can be made within the Cronobacter genus and related organism Citrobacter koseri which causes similar infections in neonates. For example, C. sakazakii genome contains 112 virulence related genes, compared with 117 in C. turicensis and 149 in Citrobacter koseri. Similarly, the C. sakazakii genome contains 179 stress related genes, compared with 178 in C. turicensis and 187 in Citrobacter koseri. Comparative genomic hybridisation using a 387,000 probe oligonucleotide tiling DNA microarray showed that of the 4,382 annotated genes on the sequenced C. sakazakii strain (BAA-894), about 55% of genes were common to all C. sakazakii strains and 43% were common to all Cronobacter spp. strains, with 1017% absence of genes. A 7 loci MLST scheme has been developed using strains which were widely distributed according to geography, time and sources. The 7 sequenced genes (atpD, fusA, glnS, gltB, gyrB, infB and ppsA) are distributed across the Cronobacter genome and give a total allelic sequence length of 3036 bp. To date 22 sequence types have been defined. The sequence typing reveals that many clinical isolates form a separate lineage to those from infant formula finished product. This may indicate that the more virulent Cronobacter strains have originated from a currently unidentified ecosystem, or may have acquired additional virulence traits from horizontal gene transfer. The MLST scheme and database is available free online at http://pubmlst.org/cronobacter/. Keywords: Cronobacter species, Enterobacter sakazakii, infant formula, genome sequencing, microarray analysis, MLST, virulence. References: Baldwin, A., Loughlin, M., Caubilla-Barron, J., Kucerova, E., Manning, G., Dowson, C. & Forsythe, S. (2009) Multilocus sequence typing of Cronobacter sakazakii and Cronobacter malonaticus reveals stable clonal structures with clinical significance which do not correlate with biotypes. BMC Microbiology 9:223. Forsythe, S. (2005) Enterobacter sakazakii and other bacteria in powdered infant milk formula. Mother and Child Nutrition. 1: 44-50. Hurrell, E., Kucerova, E., Loughlin, M., Caubilla-Barron, J., Hilton, A., Armstrong, R., Smith, C., Grant, J., Shoo, S. & Forsythe, S. (2009) Neonatal enteral feeding tubes as loci for colonisation by members of the Enterobacteriaceae. BMC Infectious Diseases 9:146. Kucerova, E., Clifton, S.W, Xia, X-Q, et al. (2010) Genome sequence of Cronobacter sakazakii BAA-894 and comparative genomic hybridization analysis with other Cronobacter species. PLoS ONE 5(3): e9556. Osaili, T., & Forsythe, S. (2009) Desiccation resistance and persistence of Cronobacter species in infant formula. Intl J. Food Microbiol. 136: 214-220. Townsend, S., Hurrell, E., & Forsythe, S.J. (2008) Virulence studies of Enterobacter sakazakii isolates associated with a neonatal intensive care unit outbreak. BMC Microbiol. 8:64.

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DETECTION OF SALMONELLA IN FOOD - EXPERIENCES AND INNOVATIONS Hanna-Leena Alakomi Saarela, M VTT Technical Research Centre of Finland, Bio- and chemical technology, P.O. Box 1000, FI-02044 VTT, Espoo, Finland

Salmonella, a genus within Enterobacteriaceae, remains as an important human pathogen. Salmonellosis has been reported to be the most common food-borne bacterial disease in the world. In the United States it has been estimated that 1.4 million non-typhoidal Salmonella infections with 400 deaths occur annually. Especially in countries with poor sanitary conditions Salmonella causes annually several deaths. Although majority of the Salmonella cases are sporadic, outbreaks occur frequently. As food trade is becoming more global and consumers prefer more fresh produce and uncooked ready-to-eat foods, the microbiological risks of imported foods have increased. The volume of global trade is increasing and food and feed stuffs are moving faster from one country to another. In addition, people are travelling more and further away than before. Hence, collaboration between various global networks/programmes needs to be reinforced. E.g. collaboration in harmonization of methods, training and organization of inter-laboratory comparison studies as well as for on-line reporting of new zoonotic cases needs to be active. The fast adaptation ability of Salmonella enhances their survival in various processes and environments. Besides the development of efficient, reliable, fast and cost-effective detection methods efforts should also be invested in Salmonella prevention in all the steps in the from farm to fork chain. Resources should also be allocated to consumer education and information about proper handling of raw food materials for prevention of cross-contamination. Salmonella has been associated with many foodstuffs. In ready-to-eat foods presence of Salmonella in considered significant regardless of the level of the contamination. Hence isolation is carried out by enrichment culture of a defined weight/volume of the food (normally 25 g). The traditional detection and isolation of Salmonella spp. from food and feed utilises a multi-step protocol with non-selective pre-enrichment, followed by a selective enrichment step, isolation on selective agar media and a preliminary biochemical and serological confirmation. Several rapid methods have been developed to speed up the detection of Salmonella. The presentation aims to give an overview of the current methodologies in Salmonella detection. Brief summary of standard methods and currently available rapid methods will be presented. In addition, the presentation gives examples of resent Salmonella outbreaks. Keywords: Salmonella, detection, outbreak References: The Community Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and foodborne outbreaks in the European Union in 2008, The EFSA Journal (2010), 1496. Voetsch et al. (2004). Clinical Infectious Diseases. 38 Suppl 3, S127S134.

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Session 7 Food additives

EMERGING AND PERSISTENT ISSUES WITH ARTIFICIAL FOOD COLOURS: METHODS OF ANALYSIS FOR NATURAL COLOUR ADDITIVES AS ALTERNATIVES TO ARTIFICIAL COLOURS IN FOOD AND DRINK Mike Scotter. The Food and Environment Research Agency, York (UK).

Concerns over the possible genotoxic and carcinogenic effects of the artificial colouring Red 2G (E128) led the European Commission, on advice from EFSA, to remove Red 2G from the authorized list in 2007 and to prohibit imports of food containing it. During 2009, in the light of the UK Southampton study on six artificial colours, EFSA lowered the ADIs for three colours and concluded that exposure of both adults and children to these could exceed the new ADIs. No grounds for changing the ADI were seen for the three other colours. While this may mean restrictions on levels or the range of foods the colours are used in, negative publicity following the study made consumers wary to their presence and UK ministers approved a voluntary ban on their use by the end of 2009. Warning labels on products containing the six colours will be mandatory in Europe, from July 2010 even though EFSA has not seen scientific substantiation for the link with hyperactivity. Natural colour additives are increasingly used as alternatives to synthetic colours in food and drink. This is partly a reaction to concerns on the safety of certain synthetic food colours which have been under intense public scrutiny for many years but is driven mainly by industry. However, natural does not necessarily mean good or safe; hence natural colours have purity specifications, usage restrictions and maximum permitted levels in line with other permitted food additives. Natural food colours include a wide range of chemical structures and physicochemical properties, so the increasing use of natural colours requires the availability of a range of suitable extraction and analytical methods for testing food and drink. This presentation will describe the state-of-the-art in this area with a review of the chemistry and analysis of the natural colours that have the greatest commercial interest.

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COMPUTER VISION BASED IMAGE ANALYSIS FOR RAPID DETECTION OF ACRYLAMIDE IN HEATED FOODS Vural Gkmen Department of Food Engineering, Hacettepe University, Beytepe, Ankara, Turkey

High concentrations of acrylamide found in common fried and baked foods attained considerable public concern since it has been classified as a probable human carcinogen. To date, many analytical methods have been published for the analysis of acrylamide in thermally processed foods. Although these methods may perform well for quality control purposes in a food analysis laboratory, they are laborious, costly, and cannot be adopted easily for process control purposes by the food industry. To satisfy the increased awareness, and greater expectation of consumers, as well as demands by the regulatory authorities, it is necessary to improve the evaluation quality of food products. Being an objective, rapid, and noncontact tool, computer vision may be a powerful technique for inspection and evaluation purposes by a rapid prediction of acrylamide level in food products. Computer vision is the construction of explicit and meaningful descriptions of physical objects from images. Image analysis is the core of computer vision with numerous algorithms and methods available to achieve the required classification and measurements. From the viewpoint of acrylamide, surface color is an important food attribute that can be used to predict the acrylamide level. It is the fact that both brown colored products and acrylamide are formed during the Maillard reaction at high temperatures . Previous findings profoundly suggest that surface color may be correlated with acrylamide in thermally processed foods. Earlier attempts were directed to measure color in CIE Lab units which is an international standard for color measurements, adopted by the Commission Internationale d'Eclairage (CIE) in 1976. Although CIE redness parameter was shown to be correlated with the level of acrylamide to a certain extent, it may not be a reliable predictor of acrylamide concentration in potato chips due to nonhomogenous surface color. This study describes a computer vision based image analysis algorithm using color segmentation for the prediction of acrylamide level in thermally processed foods. The digital images of potato chips and biscuits were used to extract a meaningful parameter to be correlated with acrylamide level. Images of processed food samples were segmented into two or three sets based on the predefined color references. Test samples (biscuits and potato chips) were analyzed using the semiautomatic segmentation algorithm to predict their acrylamide level. The results confirmed that computer vision system described here provided explicit and meaningful description from the viewpoint of inspection and evaluation purpose for potato chips and biscuits. Assuming a provisional threshold limit of 1000 ng/g acrylamide, test samples could be successfully inspected with only one failure out of 60 potato chips. The success of the algorithm was 100% on sorting biscuits based on a threshold level of 150 ng/g acrylamide. KEYWORDS: Acrylamide, potato chips, biscuits image analysis, color segmentation

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CHINA FOOD ADDITIVES HEALTH CODE(X) Zhongdong Liu 3 1 Boxiang Liu and Youning Sun
1.Henan University of Technology 2.China Food Additives & Ingredients Association 3.Tianjin Universiy To start with my reseach, lets make some introduction of Chinese Laws about food/ food additives and ingredins, regulations and standard of management system in food additive. As follows: Food hygiene law of the PRC Hygiene management measures of food additive Health standard of food additive Health standard of food nutritive fortifier General standard of food labeling policy The toxicological evaluation procedure of food safety Classification and code of food additive Classification and code of artifical flavor Declaration and acceptance regulations of food additive in chinese ministry of health National Standard of Peoples Republic of China: Organic Products This paper is in three parts: (1) Standards, Guide, Yield, The use and consumption of China Food Additives Framework (2) Risk Assessment of China Food Additives, Development of food safety standards and (3) China Food Additive Standards and international standards (Europe &MoniQA) Conclusion: The general framework of GB2760 includes two parts: General framework(format, preamble, definitions, text, etc) and Category of food additive. The use of food additives includes the following elements: range;Normative references; terms and definitions; principle of needing to use food additives; and residues of food additives. The following table show the general component elements Number Categories Nub 1 Food additives listed in Table 1 252 2 Food additives listed in Table 2 55 3 The list of foods with spices Foods with natural spices 329 Natural flavor of food equivalent 1009 Food use of synthetic fragrance 193 4 Food use of synthetic processing aids used in food industry spices 114 5 Food enzymes 44 6 Chewing Gum-based agent list 51 Total 2047 Expectation details: 1. Food additives varieties, The maximum or minimum addition; 2. In certain categories, or allow the use of certain food additives and maximum use of species (in the Food category order); 3. In a variety of food production needs in accordance with the principles of the use of additives and proper use of the list. Literature Normative Appendix 1: List of foods spices; Normative Appendix 2: the list of Food processing aid; Normative Appendix 3:the base materials and their ingredients list chewing gum; Informative Appendix 1: the functional group of Food Additives; Informative Appendix 2: the classification systems of food. ACKNOWLEDGEMENT This work was supported by the National Natural Science Foundation of China under Contract Nos.30800255, 2067262930270762209111202022081112035829576263& National 863 program of China(2007AA100401). We gratefully acknowledge for the SSRF China and comments from editors. Collaboration with Europe & MoniQA.
1.2

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AN OVERVIEW OF METHODS FOR DETERMINATION OF TRANS-FATTY ACIDS IN FOOD Mieczyslaw Obiedziski A.Obiedziska Warsaw University of Life Sciences WULS-SGGW ul. Nowoursynowska 159 02-776 Warszawa

Fatty acids containing trans unsaturated double bonds occur in nature, but the most common dietary source are man-treated fats. These in hydrogenated oils; margarines, shortenings and baked goods contain relatively high levels of trans fatty acids. Trans fatty acids appear in dairy fat because of ruminal activity, and among them conjugated linoleic acids (CLA) are a group of linoleic (18:2)-derived isomers with conjugated double bonds, mostly at carbon atoms 9 and 11 or 10 and 12. Because of the possible link between trans-fatty acids consumption and human disease mainly cardiovascular disease and cancer, the health effects of trans-FA have been studied for over the last years. On the other hand different CLA are reported to be anti-cancer, anti-atherogenic, anti-adipogenic, antidiabetogenic and/or anti-inflammatory. The analysis of fatty acids in a food matrix involves three steps: lipid extraction, preparation of fatty acid derivatives, and gas chromatographic analysis. For decades, gas chromatography (GC) has been the most frequently applied method for fatty acids analysis. Prior to GC analysis, the lipid sample has to be hydrolyzed (saponified) and fatty acids converted into non-polar derivatives, usually fatty acid methyl esters (FAMEs). This is usually obtained by direct transesterification of lipids, which proceeds more rapidly than saponification- esterification, and the reaction takes place in one step with only one reagent. However, there is some controversy over whether derivatization causes isomerization of geometrical isomers of CLAs. Furthermore the separation of a FAME mixture that contains saturated, monoenoic, dienoic, and trienoic fatty acids could be obtained by means silver ion TLC, and/or silver ion solid-phase extraction (Ag-SPE and/or silver ion column HPLC. Trans isomers, irrespective of chain length, migrate ahead of the cis isomers and are completely separated from SFAs and from dienoic acids. This technique has been extensively used prior to GC or GC/MS for the complete and more accurate analysis of geometrical isomers of fatty acids in a wide range of edible fats and oils.. The complete separation of cis and trans isomers by Ag-HPLC can be used to provide quantitative data for positional isomers not separated by GC. However, Ag-HPLC is more limited than GC as a stand-alone method for the study of geometrical and positional fatty acid isomers in fats and oils. In last years the use of comprehensive double dimensions gas chromatography combined with mass spectrometry (2D GC MS-TIC/SIM - multidimensional techniques) could be used for efficient and reliable means for quantitative determination of trans fatty acid isomers. For routine analysis, infrared spectroscopy (IR) is the simplest method for trans fat determination in oils and fats. by measurement of the absorbance of the 996 cm1 band and LOD of trans quantification of fat is usually in range 2- 5% (as % of total fat). In this paper the results of the studies on trans fatty acids contents in different food products monitored in Poland and trends of trans FA will be presented and reviewed. Key words: trans fatty acids, conjugated linoleic acid, methods of analysis

References: Trans Fatty Acids; Edited by Albert J. Dijkstra, Richard J. Hamilton, Wolf Hamm. 2008 by Blackwell Publishing Ltd. Handbook of Processed Meats and Poultry Analysis; Edited by Leo M.L. Nollet, OLLET, Fidel Toldra. 2009 by Taylor & Francis Group, LLC. Food Lipids Chemistry, Nutrition, and Biotechnology, 3rd Edition; Edited by Casimir C. Akoh, David B. Min. 2008 by Taylor & Francis Group, LLC

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Session 8 Emerging technologies for food safety assessment (BioCop, CONffIDENCE)

TARGETED AND UNTARGETED PROFILING FOR XENOBIOTICS Gaud Pinel 2 3 1 3,4 2 1 M.H. Mooney , S. Weigel , J.-P. Antignac , M.W.F. Nielen , C. Elliott , B. Le Bizec
1 1

Laboratoire dEtude des Rsidus et Contaminants dans les Aliments (LABERCA), ONIRIS, Atlanple la Chantrerie, 44307 Nantes cedex 3, France. Tel: +33 2 40 68 78 80, Fax: +33 2 40 68 78 78, e-mail: laberca@oniris-nantes.fr 2 Institute of Agri-Food and Land Use, Queens University Belfast, Stranmillis Road, Belfast BT9 5AG, Northern Ireland, United Kingdom. 3 RIKILT Institute of Food Safety, Wageningen UR, P.O. box 230, 6700 AE Wageningen, The Netherlands. 4 Wageningen University, Laboratory of Organic Chemistry, Dreijenplein 8, 6703 HB Wageningen, The Netherlands

Growth promoting practices for cattle fattening purposes are still encountered within the European Union despite the Directive 88/146/EC banning the use of growth promoters in livestock production. Detection of illegal practices classically relies on residue monitoring in a targeted approach and methods based on gas- or liquid chromatography coupled to (tandem) mass spectrometry are today considered as the state-of the-art. These strategies however fail when facing new xenobiotic growth promoting agents or new ways of application, such as the administration of low dose cocktails. In this context, screening strategies allowing detection of the physiological response resulting from anabolic compounds administration are promising approaches to detect their misuse. Profiling biological matrices to reveal biological effects of a drug can either be performed in a targeted focus on a particular class of compounds or in an untargeted way using global strategies such as transcriptomics, proteomics or metabolomics. These emerging tactics are promising ways to highlight candidate biomarkers to tackle illegal practices.

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REPLACEMENT OF THE MOUSE BIOASSAY FOR PHYCOTOXINS Luis M Botana Dept Farmacologa, Fac. Veterinaria, 27002 Lugo, Spain. Luis.Botana@usc.es

Over the past decades, the advance in the detection of marine toxins has been very slow. But the progress on the field had a major boost in recent years, due to the combination of available funds and technology. This talk will discuss the progess in the advance of alternative methods to the current legal method in Europe, the mouse bioassay. The recent progress evaluation of the EFSA Working Group on marine toxins, combined with available methods developped under the EU funded projects, BIOCOP, CONffIDENCE, DETECTOX and ATLANTOX among others, has defined a horizon of alternative options that is more than ever becoming clear. The advance in legal modifications being taken by the EU Commission is also speeding the path of change of alternative methods. All available technologies will be discussed: LC-MS-MS, receptor-based methods, antibody-based methods, and binding-based methods. The problems of toxic equivalent factors, toxin standards, and toxin groups included in the legislation, willl be also addressed. Key words: alternative methods, marine toxins, SPR, LC-MS

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VETERINARY ANTIBIOTICS, MULTIPLEX DIPSTICKS, OPTICAL AND ELECTROCHEMICAL BIOSENSORS Anne-Catherine Huet 1 1 2 2 Sara Stead , Kasia Wolodoko-Cierniak , Vincent Chabottaux , Benoit Granier , Alejandro 3 3 5 Muriano and P.Marco , , Stefan Weigel 1 Food and Enivornment Research Agency, Sand Hutton, York, North Yorkshire YO41 1LZ, uk 2 Unisensor S.A., Rue du Dossay n3, B-4020 Lige, Belgium (info@unisensor.be) 3 Applied Molecular Receptors Group (AMRg) CIBER of Bioengineering, Biomaterials and Nanomedicine CSIC, Jordi Girona, 18-26 08034-Barcelona, Spain 4 Centre dEconomie Rurale (CER Groupe), Dpartement Sant, Rue Point du Jour 8, 6900 Marloie, Belgium 5 RIKILT - Institute of Food Safety, Wageningen UR, P.O. Box 230, 6700 AE Wageningen, The Netherlands
4

Under the BIOCOP FP6 project (therapeutics work package) and the Conffidence FP7 project (antibiotics work package) a range of dipstick and electrochemical based biosensor devices as well as surface plasmon resonance (SPR) biosensor methods have been developed for the rapid detection of antibiotics in food and animal feedingstuffs. At the present time there is a requirement for the development of rapid, low-cost screening assays in order to better protect consumers and ensure EU regulations are effectively enforced. A rapid assay for monitoring low-level tetracycline residues is required to assess the effect of consumer exposure to chronic subMRL concentrations, in terms of the emergence of tetracycline resistant bacteria in the human population. The issue for honey is the presence of multiple antibiotic residues. In 2004 there were 35 RASFF alerts concerning antibiotics in honey, chloramphenicol, tylosin and sulfonamides being among the most commonly reported and the problem still persists today. The use of antibiotics in apiculture is not permitted under EU legislation. In 2003/181/EC an MRPL of 0.3 mg kg-1 for chloramphenicol in honey has been established. More recently, residues of fluoroquinolones (enrofloxacin, ciprofloxacin and norfloxacin) have been detected in Chinese honey. Within BIOCOP an immunochemical SPR biosensor screening method for fluoroquinolone (FQ) antibiotics in poultry muscle, fish and egg was developed. The assay is capable of detecting the presence of at least 13 different (fluoro)quinolones in the target matrices at low levels (LODs 0.5 50 g/kg). The applicability of the method was not only demonstrated by an in-house validation according to the European Commission decison 2002/657/EC, but also in an inter-laboratory method performance study. Competitive antibody based lateral flow dipstick assays have been developed by Unisensor. One of the new assays is the Sulfasensor which is comparable to the existing Tetrasensor. Sulfasensor can also generate rapid (< 5 min) qualitative results for multiple sulfonamides in honey at 25 g kg-1 and is applicable for point of control testing. The dipstick protocol incorporates a sample dilution step and rapid acid hydrolysis pre -treatment for the effective release sugar bound sulfonamide residues. A multiplex dipstick assay involving four test lines is also under development to detect multiple antibiotics in honey. This multiplex dipstick test is a Lateral Flow (LF) assay using both specific and generic antibodies for the recognition of chloramphenicol, tylosin and the sulfonamide and fluoroquinolone classes. The results are visualized at the 3 specific capture lines by the use of colloidal gold-conjugates. The second format of assay developed is an electrochemical immunosensor format capable of generating semiquantitative results. The immunosensor uses antigen derivatized magnetic beads and multiple-use epoxygraphite electrodes. The magnetic beads are combined directly with the sample and primary antibody, followed by the horseradish peroxidase (HRP) labeled secondary antibody to generate an amperometric signal. The assay allows detection of 11 sulfonamides in honey below a target concentration of 25 g kg-1 and is capable of analysing up to 30 samples within 3 hours. Keywords: Dipstick, electrochemical immunosensor, optical immunosensor, SPR, sulfonamides, chloramphenicol, Tylosin, fluoroquinolones, tetracyclines, honey, poultry, fish, egg.

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INNOVATIONS IN PESTICIDES ANALYSIS Jana Hajslova T. Cajka, O. Lacina, L. Vaclavk Institute of Chemical Technology, Department of Food Chemistry and Analysis, Technicka 3, 16628, Prague, Czech Republic; jana.hajslova@vscht.cz

A wide range of innovations has occurred in a pesticide residue analysis in the recent decade. High sample throughput enabled by minimum sample processing followed by fast separation / detection of target residues represent, together with achieving performance characteristics meeting legislative requirement, the main priorities in analytical research aimed at food safety control. Only recently, a new challenging technology has emerged in rapid screening of food residues. The use of ambient mass spectrometry permits the ionization of samples in the ordinary atmosphere; moreover, separation step, typically based on chromatography, and is eliminated. Generally, ambient ionization techniques can be separated into classes according to the nature of the process governing ionization. In our study, DART (Direct Analysis in Real Time), the ionization approach resembling APCI (Atmospheric Pressure Chemical Ionization) has been used for soft ionization of a wide range of both polar and non-polar residues in QuEChERS extracts obtained from cereals and other food matrices. In DART, metastable helium atoms, originated in the plasma, react with ambient water, oxygen, or other atmospheric components to produce the reactive ionizing species. I our study, the potential of DART coupled with (i) time-of flight medium resolution mass analyzer and (ii) high resolution mass analyzer employing Orbitrap technology in pesticide residue analysis will be demonstrated. The benefits and limitations OF DART-MS technique in quantitative work will be discussed. Acknowledgement: This work was realized as a part of the European Commission funded Integrated Project FOOD-CT-2004-06988 BIOCOP Key words: pesticides, residues, ambient mass spectrometry, Direct Analysis in Real Time References: 1. Schurek J., Vaclavik L., Hooijerink H., Lacina O.,Poustka J., Sharman M., Caldow M., Nielen M. W. F., Hajslova J.: Control of strobilurin fungicides in wheat using direct analysis in real time accurate time-of-flight and desorption electrospray ionization linear ion trap mass spectrometry. Anal. Chem. 80, 95679575 (2008). 2. Cajka T., Hajslova J., Mastovska K.: Mass spectrometry and hyphenated instruments in food analysis. pp. 197228. In: Handbook of Food Analysis Instruments. S. tle (ed.), ISBN -13: 9781420045666, CRC 2008, Taylor & Francis Group, Boca Raton, FL, USA

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Oral Presentations

HEAVY METAL DETECTION USING CELL BASED AND SENSOR BASED ASSAYS Danila Moscone M. Karp*, A. Rantala*, F. Arduini, J. Calvo Quintana, *Tampere University of Technology, Tampere, Finland; Tor Vergata University of Rome, Rome, Italy

Heavy-metals (HM) detection has been included both in BioCop and in CONffIDENCE EU-funded programs because of their well-documented neurotoxic, hematotoxic and nephrotoxic effects on humans. The central objective was to develop novel and accessible methods for monitoring. In the CONffIDENCE program, bacterial cells engineered by luciferase reporter genes controlled with specific genetic regulating elements, were used to determine the presence of inorganic arsenic (iAs) and methyl mercury (mHg) from fish food and feed. The sensor cells were freeze-dried to diminish batch-to-batch variation and for reagent-like usage, thus creating a permanent storage of the reporter systems. The iAs sensor had a limit of detection at 30 nM, whereas the total mercury sensor could detect mHg in the subnanomolar range. The response curves were typical bell-shaped showing the toxic levels of the metals at higher concentrations. The optimal extraction of the bioavailable portion of iAs was by simple boiling of fish tissue in H2O. Within the BioCop project, a system for lead detection has been developed, evaluated and validated for practical application in milk and baby foods. The overall approach was based on a classical electrochemical technique (anodic stripping) that was coupled with novel disposable sensors (SPEs, Screen-printed electrodes), coupled with portable, cost-effective, and user-friendly instrumentation. The final objective was to have a system for screening applicable in different settings, but which still delivered the required sensitivity for regulation. To replace the highly toxic Hg in the classic method, the SPEs have been modified by a Bi film on the surface of the working electrode. The presence of lead is monitored by i) an accumulation step in which the lead forms an alloy with bismuth, and ii) a subsequent stripping step in which a specific lead peak is observed whose area is proportional to the lead concentration. A new pretreatment method involving acid precipitation of proteins, sonication, and the addition of other agents to both enhance recovery by releasing lead from proteins, was also devised. Good recovery values in milk (93%) were achieved for detection at a concentration of 20 ppb (legal limit for lead in milk). In a parallel process, dedicated software for lead detection was developed in collaboration with the BioCop partner, PALMSENS. The software was designed to control the analytical processes but also to guide the operator through each step.The operator can provide the input data indicated by the software and have a read-out of the concentration of lead in the milk directly at the end of the procedure. Keywords: Heavy Metals; Fish; milk, cell-based biosensors; Bi-modified SPEs.

Oral Presentations

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Session 9 Authenticity and traceability

AUTHOR LOST WITHOUT TRACE: NEW ANALYTICAL APPROACHES TO TRACING THE ORIGIN OF FOOD Paul Brereton 2 and the TRACE consortium 1 The Food and Environment Research Agency 2 www.trace.eu.org
1

Food fraud and traceability continues to have a high profile with several major incidents being reported in the press and media. There has been an increased emphasis from the food industry on marketing of foods with perceived food quality attributes to an ever more discerning consumer. The lack of objective methods for verifying some of these labelling claims is to the detriment of the consumer but also the food industry, as the honest producer is not protected nor the purchasers of such products within the food chain. Many of these perceived quality attributes cannot easily be verified using current analytical methods. In particular, labelling claims that relate to: provenance, organic, identity, sustainability are difficult to substantiate and require the development of new analytical approaches and processes. Analytical methods for use in detecting food fraud rely on detecting/quantifying marker(s) of the authentic product or of the adulterant and pose considerable challenges in terms of detection, quantification and interpretation. The EU integrated project has used a multidisciplinary approach to devlop methods and systems that can trace and verify food provenance. TRACE has linked geochemistry spatial mapping, infortation technology and analytical chemistry to provide solutions in the provenencing area. Some of the latest analytical and chemometric approaches used to authenticate labelling claims will be described together with specific examples of the application of metabolite profiling methods and stable isotopic techiques for confirmation of food authenticity and traceability.

Keywords Traceability, authenticity, stable isotopes, isoscapes, trace elements, chemometrics, chemical profiling, metabolite profiling, fingerprinting, food provenance.

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Oral Presentations

ISOTOPE ANALYSIS FOR THE CONTROL OF GEOGRAPHIC ORIGIN OF TYROLEAN MILK Micha Horacek W. Papesch Environmental Resources and Technologies, Austrian Institute of Technology 2444 Seibersdorf, AUSTRIA email: micha.horacek@ait.ac.at

Consumers are willing to pay elevated prices for specific product qualities, e.g. for food from a certain region, thus there is the need to control the correct declaration of origin of these products. Conventionally this is done by estimating the flow of goods and by controlling the documentation accompanying the products. However, this means are often not sufficient to detect intentional deception. The measurement of the stable isotope composition of products offers the possibility to investigate the product itself. The stable isotope ratios of the elements H, C, N, O and S are varying geographically due to diverse environmental conditions (e.g.: climate, geology, soil, altitude, geography) thus creating individual patterns for each region. These patterns are transferred in different ways into plants and animals originating from a certain region. Therefore analysis of the stable isotope pattern can be a potent tool for geographic differentiation. Tyrolean milk is regarded as a high quality good produced under strict regulations in a special (alpine) environment. To protect Tyrolean milk from incorrectly declared milk originating from other regions/countries, stable isotope investigations have been carried out over a period of one year on samples from all regions of Tyrol. Samples have been measured for isotopic composition of H, C, N and O. The observed variation in the isotopic pattern of the Tyrolean milk within the year can be explained by different feeding regimes during summer and winter. Comparison of the isotopic pattern of the Tyrolean milk and milk samples of the same age from other regions gives evidence for significant differences in the isotope ratios. As the investigated non Tyrolean samples have been produced in neighbouring regions that sh ould have similar isotopic signals due to comparable environmental conditions, presumably it should be even easier to distinguish between milk from farther regions and milk from Tyrol. Keywords: Geographic origin, authenticity, milk, Tyrol, stable isotope, declaration of origin

Oral Presentations

Page|55

RICE TRACEABILITY SYSTEM IN TAIWAN Shin Lu a a P. Wang , R. Hsu a Researcher, China Grain Products R&D Institute, Bali, Taipei County, Taiwan, b Director, China Grain Products R&D Institute, Bali, Taipei County, Taiwan, E-mail: shin.lu@cgprdi.org.tw
b

Taiwan utilizes a voluntary Agricultural Products Traceability Certification System. This system was implemented in 2007 under the Agricultural Production and Certification Act. Since then, certain agricultural products have been certified using a label that includes product name, trace code, public information, and the Traceability Agricultural Product logo. Consumers can easily identify the traceability information through the bar code reader in the store. Being the staple food in Taiwan, rice plays an important role in the Traceability Certification System. In the east part of Taiwan, where there is a well-established rice traceability system, the rice provides higher added value in both the local market and international markets. Generally speaking, the traceability system is based on the document and record systems in farm and food processing plants. In order to use scientific evidence to identify rice origins, NIR spectroscopy and Inductively Coupled Plasma (ICP) element analysis were used to determine rice origins from the northwest, southwest, and east parts of the island. Rice varieties and planting methods were considered as well as geographic origins. Over 150 samples were collected island-wide, including japonica cultivars of Taiwan no. 2, Taiwan no. 9, and Tainan no. 11, respectively. During processing, all samples were hulled and milled under the same conditions. Brown rice and milled rice were analyzed separately using both NIR and ICP methods. Results show that NIR and ICP methods are able to distinguish significantly the rice geographic authenticity for the same rice varieties, regardless of whether the samples were brown rice or milled rice. When testing different varieties of japonica rice from different areas, the NIR method produced more accurate results for brown rice, whereas the ICP method produced better results for refined rice from different areas. Keywords: rice, food origin, authenticity, NIR, ICP Reference: Ariyama, K., H. Horita, and A. Yasui. 2004. Chemometric techniques on inorganic elements composition for the determination of the geographic origin of Welsh onions. Analytical Sciences. 20:871-877. Kim, S. S., M. R. Rhyu, et al. 2003. Authentication of rice using near-infrared reflectance spectroscopy. Cereal Chemistry 80(3): 346-349. Natsuga, M. and S. Kawamura. 2006. Visible and near-infrared reflectance spectroscopy for determining physicochemical properties of rice. American society of Agricultural and biological engineer 49(4): 1069-1076. Rittiron, R., S. Saranwong, et al. 2005. Detection of variety contamination in milled Japanese rice using a single kernel near infrared technique in transmittance mode. Journal of near Infrared Spectroscopy 13(1): 19-25. Suzuki, Y., Y. Chikaraishi, et al. 2008. Geographical origin of polished rice based on multiple element and stable isotope analyses. Food Chemistry 109(2): 470-475. Akemi Yasui and Kumiko Shindoh. 2000. Determination of the geographic origin of brown rice with trace-element composition. The Japan society for analytical chemistry 49(6): 405-410.

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Oral Presentations

IS IT ORGANIC? WHAT DO EXISTING FOOD AUTHENTICATION TECHNIQUES HAVE TO OFFER? Simon Kelly * a a a b b Mitch Kelly , Matthew Sharman , Martin Rose , Alison Bateman , Alina Mihailova , Laetitia c d d Shintu , Sren Husted & Kristian Holst Laursen a Food and Environment Research Agency, Sand Hutton, York, YO41 1LZ, UK b School of Environmental Sciences, University of East Anglia, Norwich, NR4 7TJ, UK c Aix-Marseille Universit, Campus de Saint Jrme, Av. Escadrille Normandie Nimen, 13397 Marseille cedex 20, France d Plant and Soil Science Laboratory, Department of Agriculture and Ecology, University of Copenhagen, DK * Dr Simon Kelly, Tel: +44 (0)1904 462000; E-mail: Simon.Kelly@fera.gsi.gov.uk
a

Abstract Increasing consumer demand for organic products has meant rapid expansion worldwide of the organic retail sector. Whilst the founders of the organic farming movement placed considerable value on close links between producers and consumers, the demand for organic produce has widened this gap. The organic sector is becoming increasingly dominated by corporate players that may not share some of the less tangible benefits of the organic philosophy. Notwithstanding the pros and cons of this evolution of the organic retail sector, the globalisation of organic markets must inevitably place an increased burden on certification/inspection bodies and traceability systems on which the authenticity 1 of the organic produce depends. Siderer et al. (2005) proposed the introduction of supplementary analyses and tests of organic merchandise in order to verify labelling claims. In this presentation I aim to give an overview of a range analytical techniques that have been advocated as having the potential to discriminate between different facets of organic and conventional cultivation to verify labelling claims. These include; established and validated techniques such as pesticide and veterinary drug residue analysis, stable nitrogen isotope analysis to differentiate between crops grown with the application of synthetic (inorganic) nitrogen or manure based fertiliser; trace metal analysis to establish markers for mineral supplementation or the possible effects of arbuscular mycorrhizal fungi association in organic soils; metabolite profiling, fluorescence microscopy, and other relevant techniques.

Y Siderer, A Maquet & E Anklam, (2005) Need for research to support consumer confidence in the growing organic market. Trends in Food Science and Technology. 16, 332-343.

Oral Presentations

Page|57

BIOLOGICAL BAR-CODE FOR THE DETERMINATION OF GEOGRAPHICAL ORIGIN OF FRUITS BY USING 28S RDNA FINGERPRINTING OF FUNGAL COMMUNITIES BY PCR-DGGE: AN APPLICATION TO SHEA TREE FRUITS Aly F. El Sheikha ,* 3 2 JM. Bouvet , D. Montet * Corresponding author Tel.: 33 4 67 61 57 28 Fax: 33 4 67 61 44 44 E-mail: elsheikha_aly@yahoo.com 1Minufiya University, Faculty of Agriculture, Department of Food Science and Technology, 32511 Shibin El Kom, Minufiya Government, Egypt 2Centre de Coopration Internationale en Recherche Agronomique pour le Dveloppement, CIRAD, UMR Qualisud, TA 95B/16, 34398 Montpellier Cedex 5, France 3Centre de Coopration Internationale en Recherche Agronomique pour le Dveloppement, CIRAD, UPR 39 Gntique forestire, TA A-39/C, 34398 Montpellier Cedex 5, France
1,2

Introduction: International trade intensifies and extends to the entire planet. The foodstuffs are often consumed far from their zone of production. The consumer is more and more demanding and sensitive to the quality and the origin of the foodstuffs. For long time the food industry has used simple traceability systems (El Sheikha et al., 2009) In view of the difficulties of installing these documentary systems in developing country, in particular the countries of sub-Saharan Africa, the new strategies of traceability emerge. Among the new tools of tracing the products of vegetable origin, a biological code bar based on the analysis of the DNA of micro -organisms present on the fruits is an interesting tool. Regarding Shea tree fruits, only seven countries have statistics. Nigeria accounts for more than 60% of the production of Shea butter in 2005. It is followed by Mali, Ghana and Burkina Faso, which together account for just under a third of world production in 2005 (FAOSTAT, 2007). In Europe, Shea butter is used mainly (95%) by the chocolate industry. The quantities exported to Japan, the United States or Switzerland would be mainly used for cosmetic or pharmacological (UNCTAD, 2001). Purpose: A molecular technique employing 28S rDNA profiles generated by PCR-DGGE was used to detect the variation in fungal community structures of Shea tree fruit (Vitellaria paradoxa) from Ghana, Senegal, Mali and Cameroon. Results: When the 28S rDNA profiles were analyzed by multivariate analysis, distinct microbial communities were detected. The band profiles of Shea tree fruit fungi from different countries were specific for each location and could be used as a bar code to discriminate the origin of the fruits. Significance of study: This method is a new traceability tool which provides fruit products with a unique biological bar code and makes it possible to trace back the fruits to their original location. Keywords: traceability, PCR-DGGE, geographical origin References El Sheikha, A.F., Condur, A., Mtayer, I., Le Nguyen, D.D, Loiseau, G., and Montet, D. 2009. Determination of fruit origin by using 26S rDNA fingerprinting of yeast communities by PCR-DGGE: Preliminary application to Physalis fruits from Egypt. Yeast 26 (10): 567-573. Food and Agriculture Organization of the United Nations Statistics FAOSTAT. 2007. Shea Tree Fruits Production and Distribution in all of the World. UNCTAD. 2001. United Nations Conference on Trade and Development. Organ of the UN. UNCTAD secretariat based on the statistics of the United Nations Food and Agriculture. Market information in the field of commodities Shea Tree Fruits, N 2885. Available at: http://unctad.org/infocomm/francais/karite/plan.htm. Accessed February 2001.

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Oral Presentations

POSTER PRESENTATIONS
Nr Last name First name Organisation Abstract title Detection of Mustard Residue Contamination in Food Commodities by Immuno Assay-Based Test A Sensitive, Rapid, ELISA Test for the Detection and Quantitation of T-2 and HT2 Toxins in Grain Commodities Shelf Life Prediction of Biscuit Products using Critical Moisture Content and Moisture Sorption Model Detection of Bacillus cereus toxins Chemical and microbial contaminants in baby foods available in the Portuguese market Traceability and Food Safety of Poultry Meat Microbiological, Chemical, and Sensory Quality of Mackerel (Scomber scombrus) Fish during Iced Storage Mitigation of Acrylamide Formation during Malt Processing Implementing the system of traceability on the wine chain Development of Reference Material for Gluten Quantification H, C, N and S stable isotope ratios and elemental data for the traceability of Italian PDO cheeses The influence of post-harvest storage conditions upon the microbiological contamination on peach and nectarin fruits Comparison of Acrylamide Content in Breads Differing in Formulas Method development for the detection of potentially allergenic fining agents in white wine: lysozyme and ovalbumin Effect of baking conditions on crust coloration and acrylamide formation during baking of white bread Universal Biological Bar-Code for Determining the Geographical Origin of Fruits by Using PCR-DGGE The distribution of fungi and mycotoxins in South African maize mills Validation of Analysis Method of Cu Metal on Canned Food With AAS (Atomic Absorption Spectroscopy) Instrument A novel high-resolution mass spectrometry strategy for the non-targeted detection of chemicals in food Effect of Transglutaminase Enzyme and Maillard Reaction on the Loss of Lysine in Crackers Allergen management: tracking down hidden allergens with lateral flow rapid tests Poster Presentations Page|59 Session

P1

Abouzied

Mohamed

Neogen

food allergens chemical contaminants

P2

Abouzied

Mohamed

P3 P4

Adawiyah Alakomi

Dede HannaLeena

Neogen IPB (Institut Pertanian Bogor) VTT National Health Institute Doutor Ricardo Jorge UFT University of Plymouth HCTU ASE BUTE IASMA Fondazione E Mach

microbiology microbiological contaminants mycotoxins & phycotoxins authenticity & traceability microbiological contaminants food additives authenticity & traceability food allergens authenticity & traceability microbiological contaminants food additives

P5 P6

Alvito Angelova

Paula Galena

P7 P8 P9 P10

Banja Basinci Boboc Bugyi

Bamba Fatma Dan

Zsuzsanna

P11

Camin

Federica

P12 P13

Chira Ciesarova

Adrian Zuzana

UASVM VUP

P14

Corrales

Margarita

P15

Dessev

Tzvetelin

JRC IRMM University of Food Technologies Minufiya University University of Pretoria

food allergens

food additives authenticity & traceability mycotoxins & phycotoxins chemical contaminants chemical contaminants

P16 P17

El Sheikha Erasmus

Aly Corinda

P18

Faridah

Didah Nur

IPB Thermo Fisher Scientific

P19

Godula

Michal

P20

Gumus

Seher Christine Maria

HCTU

food additives

P21

Gutschelhofer

R-Biopharm AG

food allergens

Nr

Last name

First name

Organisation

Abstract title Establishment of a lab made magnetic beads and multiplex PCR system for the simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella Extraction of Wheat Commodities With Different Extraction Buffers and Their Influence on Signal Intensity in TripleQuadrupole Mass Spectrometry Inhibition of Escherichia coli by the lactic acid bacterium Lactobacillus plantarum B33 in mustard leaves fermentation Molecularly imprinted solid-phase extraction combined with high performance liquid chromatography for determination of furan in instant coffee Priorities and needs for implementation of updated HACCP systems and modern monitoring technologies at the Bulgarian food sector The analysis of status of the standards system of food contact plastic materials in China Acrylamide Content in Cookies Enriched by Fruit Fibre Characterisation and Classification of Serbian Honey by Physicochemical Parameters Implementation of HACCP systems and rapid methods in the food industries Microbiological and Hygienic Aspects of Pink-Colored Non Scabby Kernels Resistance to ciprofloxacin and tetracycline of Campylobacter spp. isolated from poultry products in Poland Occurrence of Campylobacter spp. in poultry products for sale on the Polish retail market The Effect of Prebiotic Addition and Incubation pH to Synbiotic Soyghurt Dietary Exposure of Acrylamide in High School Student Group Evaluation of yeast contamination sources in traditional Iranian yoghurt drink (Doogh) Cereals and cereal products authenticity issues Antimicrobial activity of supercritical fluidcarbon dioxide hop extracts Simultaneous Determination of Deoxynivalenol, Zearalenone, T-2 & HT-2 Using DZT MS-PREP in Conjunction With LC-MS/MS Trans fatty acids level of frying fats in Poland Determination of acrylamide in coffee by LC-MS/MS and estimation of dietary exposure to acrylamide from coffee in Polish population

Session

P22

Haojiang

Zuo

Sichuan University

microbiological contaminants

P23

Heick

Julia

Eurofins Hanoi University of Technology

food allergens microbiological contaminants

P24

Ho

Phu-Ha

P25

Huang

Xue-Song

Jinan University

food additives

P26

Ivanova

Mirena

UFT Sichuan University VP Center for Food Analysis NTUA VNIIZ

MoniQA outputs/general MoniQA outputs/general food additives authenticity & traceability MoniQA outputs/general microbiological contaminants microbiological contaminants microbiological contaminants microbiological contaminants food additives

P27 P28

Jinyao Kukurova

Chen Kristina

P29 P30 P31

Lazarevic Lebesi Machikhina

Kristina Dimitra Lidia

P32

Mackiw

Elzbieta

NFNI

P33 P34 P35

Mackiw Manullang Markova

Elzbieta Monang Lucie

NFNI IPB VP Food Science and Technology Institute INRAN UFT

P36 P37 P38

Mehraban Melini Mihaylova

Masoomeh Francesca

Dasha

microbiology authenticity & traceability microbiological contaminants

P39 P40

Milligan Mojska

Claire Hanna

R -Biopharm Rhone NFNI

chemical contaminants food additives

P41

Mojska

Hanna

NFNI

food additives

60|Page

Poster Presentations

Nr

Last name

First name

Organisation

Abstract title Identification of peptide markers for caseinate by LC- ESI - high resolution MS: application to fined white wines Dynamics and Biodiversity of microorganisms (fungi and yeast) by PCRDGGE, with the objective of understanding OTA production on coffee beans Evaluation of the Extraction Procedure for the Determination of Deoxynivalenol in Maize Investigation of the various derivatization methods including trimethylsilylation of melamine, ammeline, ammelide and cyanuric acid for the simultaneous GC-MS analysis POPs (Persistent Organic Pollutants) in Some Fish Species From Thyrrenian and Adriatic Sea Validation protocols for food analysis methods. Measurement uncertainty and validation criteria Application of Cellular Toxicity Analysis Method in vitro on Red Fruit (Pandanus conoideus Lam) Extract Risk of accumulation of attendant pharmaceutical residues in waste waters for fish meat ICC Your Connection to the World of Cereals QAS Quality Assurance and Safety of Crops & Foods Rapid Methods for food additives: present status, gaps and needs! Survey: Ochratoxin A in wines (VQPRD) from the North Western Greece (Achaia, Ileia) Survey of Phthalate Levels in Food on the Belgian Market Ochratoxin-A Producing Moulds and Ochratoxin A in Forages Development and validation of an immunoaffinity based HPLC method for fumonisin determination in maize-based baby food products Type and Concentration of Coagulant Effect Texture Sensation of CoagulatedBased Product Traditional fermented sausages: Microbiological risks and potential solutions Quality and Safety of Food in Mass Catering in Polish Hospitals - the Assessment of Realization of Hygienic Standards and HACCP System Implementantion Food Allergens and HACCP: Hazards Induced and Additional Controls Verification of the identity of fats and oils Poster Presentations Page|61

Session

P42

Monaci

Linda

ISPA - CNR

food allergens

P43

Montet

Didier

CIRAD

mycotoxins & phycotoxins mycotoxins & phycotoxins

P44

Numanoglu

Eren

HCTU

P45

OH

ChangHwan

HAITAI Confectionery & Foods Co., Ltd

chemical contaminants chemical contaminants MoniQA outputs/general

P46

Orban

Elena

INRAN

P47

Oreopoulou

Vassiliki NurheniSri

P48

Palupi

NTUA Bogor Agricultural University

food additives chemical contaminants MoniQA outputs/general MoniQA outputs/general food additives mycotoxins & phycotoxins chemical contaminants mycotoxins & phycotoxins

P49 P50 P51 P52

Papp Poms Poms Psimouli

Zsuzsanna

HAKI ICC ICC NTUA Toxin Analysis Lab ELETOX VITO NV University of Novi Sad

Roland Roland Vassiliki

P53 P54 P55

Sarigiannis Servaes Skrinjar

Yiannis Kelly Marija

P56

Solfrizzo

Michele

P57

Syah

Dahrul

CNR ISPA Bogor Agricultural University Hanoi University of Technology

mycotoxins & phycotoxins

food additives microbiological contaminants

P58

Tran

Khanh

P59 P60 P61

Turlejska Tzia van Ruth

Halina C. Saskia

NFNI NTUA RIKILT

MoniQA outputs/general food allergens authenticity & traceability

Nr

Last name

First name

Organisation

Abstract title A world trade market dilemma of rapeseed (seed-oil); a countervailing risk of a synergistic interaction between aflatoxin B1 and hepatitis B/C, leads to a significant health risk and food-borne illness, is studying vs. the economic impact in China and EU The occurrence of false positive samples in determination of OTA in herbal tea, spices and dietetic products New reliable method using MycoSep 150 clean up column to detect ergot alkaloids in agricultural commodities Detection of toxic fragments from gluten using a new monoclonal antibody-based test Comparative Study on Safety Assessment for Flavourings of CAC, United States, European Union and China Primary Study on the Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Staphylococcus aureus

Session

P62

Vered

Micha

P63

Vukovic

Gorica

P64

Wanzenboeck

Eva

P65

Wanzenboeck

Eva

VTAG Institute of Public Health, Belgrade Romer Labs Diagnostic GmbH Romer Labs Diagnostic GmbH Sichuan University Sichuan University

mycotoxins & phycotoxins mycotoxins & phycotoxins mycotoxins & phycotoxins

food allergens

P66

Xiao-Fen

Yao

food additives microbiological contaminants

P67

Zhimei

Xie

62|Page

Poster Presentations

P1 DETECTION OF MUSTARD RESIDUE CONTAMINATION IN FOOD COMMODITIES BY IMMUNO ASSAY-BASED TEST. M. Abouzied M. Sarzynski Neogen Corporation, Lansing, Michigan, USA Allergic reactions to mustard, including severe anaphylaxis, have been reported. Mustard is one of the listed allergen in EU labeling directives that food industry required to test for its presence in final products. A 30-min Mustard Allergen Test for quantitative analysis of mustard contamination in finished food products and in incoming raw ingredients was developed. The test is a Sandwich Enzyme-Linked Immunosorbent assay (S-ELISA). Polyclonal antibodies against specific mustard protein markers were used as the capture and detector antibodies. For quantitative analyses, a standard curve of mustard flour ranging from 0 to 25 PPM (0 to 1 g ml) was used. Samples were extracted by shaking 5 g of ground samples with 125 ml of extraction buffer (Tris/EDTA) in a hot water bath. Extracts were filtered and filtrates were used directly for ELISA analyses. Extracts are added to antibody-coated wells in which mustard proteins bind to the capture antibody during a 10-min incubation period. Any unbound protein is washed away and anti-mustard horseradish peroxidase-labeled antibody (detector antibody) is added. The detector antibody binds to the mustard protein during a 10-min incubation period. Unbound enzyme-labeled antibody is washed away and a substrate is added. Color develops as a result of the presence of bound-labeled antibody during a final 10-min incubation period. Absorbance readings of samples are compared with those of the standards and the concentrations in parts per million (PPM) are calculated. Mean recovery of mustard from various spiked samples was found to be 88%. Limit of detection was found to be less than 2.5 PPM (0.1 g/ml) determined as mustard flour. The test is capable of detecting residual contamination of mustard in ingredients, finish products and rinses and on environmental surfaces. No cross-reactivity was observed with any other spices, plant or animal proteins. The test can be used to detect inadvertent mustard contaminations in ingredients, finished products such as flours, tomato products, spices, salad dressing, meat products, and bakery products. KEYWORDS: Mustard, allergen, Sandwich ELISA

Poster Presentations

Page|63

P2 A SENSITIVE, RAPID, ELISA TEST FOR THE DETECTION AND QUANTITATION OF T-2 AND HT2 TOXINS IN GRAIN COMMODITIES M. Abouzied A. Walsh Neogen Corporation, Lansing, MI, USA

T-2 toxin and HT-2 toxin are potent toxins belong to the group trichothecenes type A mycotoxins that usually produced by fungi of the Fusarium genus, which are commonly found in various cereal crops (wheat, corn, barley, oats, and rye) and processed grains (malt, beer and bread). T-2-and HT-2 toxins often occur together in infected cereals. They elicit a severe inflammatory reaction in humans and animals and have teratogenic effects. We developed an easy to use polyclonal antibody-based Enzyme Linked Immunosorbent Assay (ELISA) for the detection and quantification of T-2 and HT-2 toxins in agriculture commodities. The assay is a direct competitive ELISA in a micro-well format. The detection limit of the assay is less than 1 ng/ml (less than 10 ppb) of individual T-2 and HT-2 or a mixture of both mycotoxins. The test can detect T-2 or HT-2 at 100% of either or a mixture of them in corn, wheat, barley, rye, soybean and oats. Concentration of T-2 or HT-2 required for 50% binding inhibition is 6.8 ng /ml (68 ppb). The antibody used is very specific for T-2 and HT-2. The antibody has no cross reactivity with other trichothecene mycotoxins such as T2-tetraol, deoxynivalenol (DON), 3acetyl-DON, 15-acetyl-DON, fusarenon-x, zearalenone or nivalenol. Samples were extracted by shaking 5 g ground sample with 25 ml of 70% methanol-water (1:5) for 3 min. Extracts were diluted 1:1 with water and then used in the ELISA test. The mean recovery of T-2, HT-2 or a mixture of both in corn, wheat, barley, soybean and oats determined by this method was 87%. The test correlates very well with a reference method. Correlation data between the CD-ELISA and LC/MS method yielded a correlation coefficient (r value) of 0.995. This assay may be used to quantitate T-2 / HT-2 in food and feed samples within 10 minutes. KEYWORDS: T-2, HT-2, Direct Competitive ELISA, cross-reactivity

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Poster Presentations

P3 SHELF LIFE PREDICTION OF BISCUIT PRODUCTS USING CRITICAL MOISTURE CONTENT AND MOISTURE SORPTION MODEL D. Adawiyah F. Kusnandar and M. Fitria Department of Food Science and Technology, Faculty of Agricultural Technology, Bogor Agriculture University, Indonesia. Email: dede_adawiyah@yahoo.com

The objective of this research was to predict the shelf life of two type commercial biscuit product (soft and hard biscuit) using Accelerated Shelf Life Testing (ASLT) method applying critical moisture content model and moisture sorption isotherm curve according Labuzas equation. This research also evaluated the effect of packaging material and relative humidity to the shelf life of commercial biscuit products. Critical moisture content was determined by stored biscuit sample at relative humidity (RH) 75% and evaluated the crispiness periodically using texture analyzer. The soft biscuit had critical moisture content when the percentage decline of crispiness was 60%, while hard biscuit was 50%. Moisture sorption isotherm was conducted at five equilibrium relative humidity using five o saturated salts (30 C) using GAB (Guggenheim, Anderson, de Boer) model. Water vapor transmission rate of packaging material (polypropylene plastic and metalized plastic) was determined by ASTM, F1249-01 using Permatran Mocon W*3/31. The type of packaging material very affected the predicted shelf life value. Hard biscuit packed with metalized plastic was predicted had shelf life 15.6 months (16.4 months for soft biscuit), while that of packed with polypropylene packaging was predicted had shelf life 3.8 months (3.0 months for soft biscuit) Type of biscuits did not have a significant influence on predicted shelf life of biscuits. The higher relative humidity or the higher storage temperature, the shorter the product shelf life.

Keywords : shelf life, accelerated method, biscuit, critical moisture content

Refferences: nd Bell, L. N. Dan Labuza, T. P. 2000. Moisture Sorption Practical Aspects of Isotherm Measurement and Use 2 Edition. American Association of Cereal Chemists, Inc., USA. Boente, G., Gonzalez, H. H.L., Martinez, E., Pollio, M. L., dan Resnik, S. L. 1996. Sorption isotherm of corn-study of mathematical models. J. Food Eng. 29:115-128. Brown, E. W. 1992. Plastic in Food Packaging, Properties, Design, and Fabrication. Marcell Dekker Inc., New York. Chirife, J. dan Iglesias, H. A. 1978. Equation for fitting water sorption isotherm of foods. Part I a review. J. Food Tech. 13:159-593. Ellis, M. J. 1994. The Methodology of Shelf LifeDetermination. Di dalam : Shelf Life Evaluation of Foods. C. M. D. Man dan A. A. Jones, hal 27. Blackie Academic & Professional, London. Eskin, M. dan Robinson, D. 2001. Food Shelf Life Stability: Chemical, Biochemical and Microbial Changes. CRC Press, USA. Gunasekharan, V. dan John, D. F. 1993. Shlef Life Prediction of Packaged Foods. Di dalam : Shelf Life Studies of Foods and Beverages. Charalambous, G. (ed). Elsevier, New York. Labuza, T. P. 1982. Shelf Life Dating of Foods. Food and Nutrition Press., Inc., Westport, Connecticut. Manley, D. J. R. 1983. Technology of Biscuits, Crackers, and Cookies. Ellies Horwood Ltd. Publ., England. nd Matz, S. A. dan Matz, T. D. 1978. Cookie and Crackers Technology 2 Edition. AVI Publishing. Co. Inc., Westport. Mir, M. A. dan Nath. 1995. Sorption isotherm of fortified mango bars. J. Food Eng. 25:141-150. Reardon dan Wade. 1991. Some Water Based Adjuncts. Di dalam: Biscuits, Cookies, and Crackers. AVI Publishing Company, Connecticut. Sun, W. D. 2000. Comparison and Selection of EMC/ERH Isotherm Equation for Drying and Storage of Grain and Oilseed. University College Dublin, Dublin.

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P4 DETECTION OF BACILLUS CEREUS TOXINS H-L. Alakomi M. Kivilompolo, T. Seppnen-Laakso, T. Hytylinen, Saarela, M. & A. Laitila VTT Technical research centre of Finland, Bio- and chemical technology, P.O. Box 1000, FI-02044, VTT, Finland

Bacillus cereus is a facultatively anaerobic, spore-forming bacterium and often associated with food contamination, food spoilage and food poisoning. Elimination of spores from food processes is difficult, since spores are highly heat- and UV-resistant as well as more tolerant to disinfecting agents than vegetative cells. B. cereus is the causative agent of two distinct forms of gastroenteritic disease connected to food poisoning. B. cereus produces an emesis-causing toxin, cereulide, and three enterotoxins that cause diarrhea. A close genetic relationship has been observed between all members of B. cereus group- which currently includes six species, namely B. anthracis, B. cereus, B. mycoides, B. pseudomycoides, B. thuringiensis and B. weihenstephanensis. Enterotoxin production is broadly distributed among different members of B. cereus group and also reported in other Bacillus sp., whereas emetic toxin formation has been associated to certain group of B. cereus. The emetic toxin, cereulide, is heat- and acid-stable and toxic at low doses. Bacillus food poisoning usually occurs because spores survive cooking or pasteurization and then germinate and multiply when food is inadequately refrigerated. The emetic type of food borne disease is often connected to rice, pasta and other cereal-based products. The increased production of readyto-eat foods with prolonged storage periods and mild processing methods is an increasing risk for cereulide production. Hence, verification methods for toxic microbial metabolites are needed to assure food safety. The aim of this study was to develop a method for detection of cereulide production in Bacillus isolates and in food products. In addition, enterotoxins were determined with commercial assays. For determination of cerulide, its two homologs and valinomycin, a high-throughput method based on ultra high performance liquid chromatography connected to triple quadrupole mass spectrometry (MS) was developed. Cereulide was also determined from spiked food matrix samples (cereal products). In addition, cereulide toxin genes were verified from the inoculated cells with a commercial kit. Enterotoxin production by known toxin producing strains from culture colonies and in liquid cultures was followed by commercial immunological detection assays. The applied methods provided fast assays for detection of toxigenic isolates and emetic cereulide toxin from suspected Bacillus colonies and food matrix samples.

Keywords Bacillus cereus, toxin, cereulide, UHPL-MS

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P5 CHEMICAL AND MICROBIAL CONTAMINANTS IN BABY FOODS AVAILABLE IN THE PORTUGUESE MARKET P. Alvito C. Martins, E. Vasco, S. Santiago, R. Furtado and M.A. Calhau. Food and Nutrition Department, National Health Institute Doutor Ricardo Jorge, Av. Padre Cruz, 1649016 Lisboa, Portugal; paula.alvito@insa.min-saude.pt Food can be contaminated with chemical or microbiological agents and children are especially vulnerable to acute, sub-acute and chronic effects of ingestion of food hazards (Vracko et al, 2007). The present study aims to study the occurrence of food contaminants in infant formula and follow-on formula (milk powder), processed-cereal based foods (flours), apple- and vegetable-based baby foods available in the Portuguese market during the period between 2007 and 2008. A total of 167 samples labelled as being intended for infants and young children were purchased in Lisboa city and analysed for mycotoxins (patulin, aflatoxins M1 and B1 and ochratoxin A), nitrates and heavy metals (Pb, Cd and Hg). Infant formula, follow-on formula and processed cereal-based foods were further analysed for microbiological contamination, namely, food safety and process hygiene criteria. Mycotoxins and nitrates were analysed by HPLC and toxic metals by Atomic Absorption Spectrometry (Pb and Cd) and a Direct Mercury Analyser DMA80 (Hg). Validation criteria were evaluated prior to sample analysis. Validated methods and ISO standards were used for microbiological determinations. The mycotoxins patulin, AFM1, AFB1 and OTA were quantified in 2 of 86 (2/86) samples, 4/27, 1/27 and 10/27 samples with maximum values of 5.6, 0.041, 0.009 and 0.212 g/kg, respectively (Barreira et al, 2010; Alvito et al, 2010). Nitrates were quantified in 53 of 80 (53/80) samples with a maximum value of 240 mg/kg. The toxic metals Pb, Cd and Hg were quantified in 6/16, 6/9 and in 38/81 samples with maximum values of 63, 69 and 13 g/kg, respectively. When available, results were compared with legislated values. According to the Regulation (CE) 1881/2006 of 19 December, all samples revealed levels below the legislated values except for one sample detected in vegetablebased baby foods with high nitrates (maximum legislated values 200 mg/kg). This fact could indicate the need to analyse further vegetable-based baby foods. No values were established for Pb and Cd in processed-cereal foods and for Hg in baby foods for infants and young children. Forty eight samples (29 cereals and 19 infant formula) were analysed for microbiological assessement. The pathogenics Salmonella and Cronobacter sakazakii were not detected. The presence of Enterobactereaceae in 4 samples reveals post heat-treatment contamination. Only low numbers of Bacillus cereus ( <100 colony forming units cfu/g) were found; its presence in 10 samples and the aerobic plate counts > 104 cfu/g suggests the need of a better selection of raw material and improvements in production hygiene in these type of foodstuffs. This is the first study reporting levels of food contaminants in baby foods available for consume in the Portuguese market and one of the few monitoring studies on baby foods contamination. The present results suggest that baby foods have a good sanitary quality for chemical contaminants and its consumption is safe.

Keywords: baby foods, mycotoxins, nitrates, heavy metals, microbial criteria, food safety References: -Alvito, P.C., Sizoo, E.A. Almeida, C.M. M. and Van Egmond, Hans P.(2010). Occurrence of Aflatoxins and Ochratoxin A in Baby Foods in Portugal, Food Analytical Methods, 3,1, 22 30. -Barreira M.J., Alvito P. C. and Almeida C.M.M. (2010). Occurrence of Patulin in apple-based foods in Portugal. Food Chemistry 121, 3: 653-658. -Vracko P., Tuomisto J., Grad J., and Kunseler E. (2007). Exposure of children to chemical hazards in food. European Environment and Health Information System. Geneva, World Health Organisation.

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P6 TRACEABILITY AND FOOD SAFETY OF POULTRY MEAT G. Angelova D.Yordanov, M. Ivanova, A. Angelov University of Food Technology, 26 Maritza Blvd., 4002 Plovdiv, Bulgaria

Traceability is increasingly becoming standard across the agri-food industry, largely driven by recent food crises and the consequent demands for transparency within the food chain. Traceability also forms an essential component of any risk management strategy and is a key requirement for postmarketing surveillance. Traceability must aim to create a link between the various steps in the entire food chain. These steps must cover the animal production at the farm, processing in meat plants and other food premises, distribution to wholesalers and retailers and right through to the moment the food is placed on the consumers table. Tracing can provide greater confidence in certification schemes, especially regarding their disease-free status. Most poultry products are produced by companies who control all facets of production. The primary breeding, commercial breeding and production stages of poultry production have had to develop comprehensive recording and traceability systems. These systems are principally documentary trails on an individual flock basis. Each flock comprises a unit having the same or similar status receiving a unique lot number. Individual identification of poultry is not generally practiced commercially. Lately, a new approach of combined traceability is being discussed in which the product flow should be accompanied by information about the microbiological status that is related with a food safety. Campylobacter spp., the commonest cause of food poisoning, is strongly associated with poultry. Contamination occurs both on the farm and in poultry slaughter plants. At the plant, de-feathering, evisceration and carcass chiller can lead to higher microbial risks due to the possible crosscontamination. To improve the quickly typing and quantifying of this pathogen in the processing plant the new molecular methods are being explored. The aim of our work is divided in two sections, the first, to provide an overview of the steps of poultry processing and its influence on product safety, the second, to focus on the application of a combined traceability of poultry products during the processing together with implementation of new molecular methods for Campylobacter spp. analyses. Key words: Traceability, food safety, Campylobacter spp., poultry products. References Atterbury R., Connerton Ph., Dodd C., Rees C., Connerton I., 2003, Isolation and characterization of Campylobacter bacteriophages from retail poultry, Applied and Environmental Microbiology, Aug. 4511-4518 Fallon M., 2001, Traceability of poultry and poultry products, Rev. sci. tech. Off. Int. Epiz., 20(2), 538546 Keener K., Bashor M., Curtis P., Sheldon B., Kathariou S., 2004, Comprehensive review of Campylobacter and poultry processing, Comprehensive reviews in food science and food safety, Vol.3, 105-116 Yordanov D., Angelova G., 2006, Identification and traceability of meat and meat products, Biotechnol&Biotechnol Eq., 20(1), 3-8

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P7 MICROBIOLOGICAL, CHEMICAL, AND SENSORY QUALITY OF MACKEREL ( SCOMBER SCOMBRUS) FISH DURING ICED STORAGE B. Banja G. Bradley, W. AL-Murrani and V. Kuri School of Biomedical and Biological Sciences, University of Plymouth, PL4 8AA, England, UK E-mail: Bamba.Banja@plymouth.ac.uk

ABSTRACT Microbiological, chemical, and sensory analyses were conducted on mackerel fish over 17 days to determine the quality changes during iced storage. Sensory evaluation was performed using the quality index method (QIM); for the determination of volatile bases (total volatile bases nitrogen-TVBN and trimethylamine-TMA) steam distillation was carried out using a Kjeldahl distillator. Microbiological evaluation was performed for bacterial counts using tryptone soya agar (TSA), iron agar (IA) and milk agar (MA). Sensory results indicated that whole un-gutted mackerel fish had a shelf life of 8 days. The quality index (QI) was highly correlated (r=0.99) between the average QI and days of storage. Both TVB-N and TMA increased from 8.42 to 63.73mgN/100g and 4.35 to 25.41 mgN/100g respectively during icing storage. The production of TMA was related to TVB-N and this relationship was highly significant (r= 0.97; p< 0.001). The regression of TVB-N on TMA was only highly significant (p< 0.001) for day 8. A highly significant increase in bacterial counts over the days of iced storage (p< 0.001) for all the species of microorganisms enumerated on the different culture media. A major increase in mean microbial count occurred at day 8; the final cumulative increase at day 14 was over 6 fold compared to day 3. This indicated that the usefulness of total bacterial count, as an indicator of spoilage, is only clear from day 8. The 95% confidence interval (CI) multiple comparisons confirmed the same trend for the other bacterial groups. Keywords: Microbiological analysis, sensory evaluation, fish freshness, shelf life, quality of mackerel, quality index method

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P8 MITIGATION OF ACRYLAMIDE FORMATION DURING MALT PROCESSING F. Basnc 1 2 1 1 B.A. Mogol , S. Gler , V. Gkmen , H. Kksel * 1 Food Engineering Department, Hacettepe University, Beytepe, Ankara, Turkey 2 Central Field Crop Research Institute, Yenimahalle, Ankara, Turkey
1

Abstract Acrylamide is classified as a probable human carcinogen by IARC (1994) and detected in a wide range of processed carbohydrate rich foods by Swedish researchers in 2002 (Tareke et al., 2002; SNFA, 2002). Dark roasted malts are generally used in various beverages and foods to obtain aroma and dark color but contain significant levels of acrylamide (Mikulkov and Sobotov, 2007). Hence, new strategies are needed to mitigate acrylamide formation during malt production. In this study, precursors of acrylamide formation, effect of roasting temperature and mitigation strategies for acrylamide formation during malt processing were examined. The barley with a good malting quality (cv. Atilir) was steeped for 34 h and germinated for 110 h in an automatic micro-malting system (Danbrew, Denmark) at 15C. Green malt was dried in drying oven (Memmert, Germany) for 16 h at 50C. Acrylamide content was determined at the temperatures between 60-200C. For the restriction, asparaginase (3000 and 4000 ASNU/kg) and/or glycine (4000 and 5000 ppm) were applied to green malt before kilning and the samples roasted at 150, 175 and 200C for 2 h, separately. Asparagine, reducing sugar (Palazolu and Gkmen, 2007), acrylamide contents (enyuva and Gkmen, 2005) and color values (CIE L*a*b*) (Gkmen and St, 2007) were determined. Statistical analyses were carried out by using SSPS Data Editor 15.0 with One-Way ANOVA. The results indicated that reducing sugar and asparagine levels of barley generally increased during germination. Acrylamide content increased and the color became darker as the roasting temperature increased. Acrylamide content decreased around 20 and 32% by the application of the enzyme at the levels of 3000 and 4000 ASNU/kg green malt, respectively. Acrylamide levels decreased around 16 and 22% by the application of glycine on green malt at the levels of 4000 and 5000 ppm, respectively. With the application of enzyme and glycine in combination, the decrease in acrylamide content was around 51%. The colors of dark roasted malts were not negatively influenced from enzyme and glycine treatments. It can be concluded that dark roasted malts with significantly lower acrylamide content can be produced without deterioration in color. Keywords: Malt, acrylamide, Maillard reaction, barley, asparaginase, glycine References: IARC (International Agency for Research on Cancer), 1994, Acrylamide, Lyon, France. Tareke, E.; Rydberg, P.; Karlsson, P.; Eriksson, S.; Trnqvist, M., 2002, J. Agr. Food Chem., 50, 4998-5006. SNFA (Swedish National Food Administration), 2002. http://www.slv.se/en-gb/ Mikulkov, R. and Sobotov, K., Acta Chim. Slov., 2007, 54, 98 101. Palazolu, T.K. and Gkmen V., 2008, J. Agr. Food Chem., 56, 6162-6166. enyuva, H.Z. and Gkmen, V., 2005, Food Chem., 97, 539-545. Gkmen, V. and St, , 2007, J. Food Eng., Vol.3, Iss.5, Article 5.

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P9 IMPLEMENTING THE SYSTEM OF TRACEABILITY ON THE WINE CHAIN D. Boboc A. Tudorache, R. Ion, M.Bran Bucharest Academy of Economic Studies, Faculty of Agro-food and Environmental Economics

Abstract The paper shows the results of a scientific research referred to the implementation of traceability system in the wine chain in Romania. It answers the questions how this system is implemented and whether its implementation brings transparency in doing operations within the wine chain. In pursuing these questions, there are presented the scheme of implementing the system of traceability on the wine chain, including its activities, reports and documentation needed, and its implementation to the white wine. The results show that the implementation of the system of traceability on the wine chain brings higher performance in the wine agro-food system. Keywords: traceability, wine chain, authenticity, wine labeling

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P10 DEVELOPMENT OF REFERENCE MATERIAL FOR GLUTEN QUANTIFICATION Zs. Bugyi K. Trk, L. Hajas, Zs. Adonyi, S. Tmskzi Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, H-1111 Budapest, Megyetem rkp. 3.

Proteins of different cereals belonging to the prolamin superfamily are responsible for triggering certain adverse reactions against these cereals, mainly wheat (Breitender, 2004). These hypersensitivity reactions can be classified as intolerances or allergies. The best-known cereal related illness is coeliac disease which is provoked by the proteins of gluten. The prevalence of wheat allergy is 20% within the allergic population and 1% of the population is affected by coeliac disease (Hischenhuber, 2006). The only effective treatment of these illnesses is a diet with the total avoidance of the allergenic proteins (Gendel, 2008). To ensure the safety of people suffering from these reactions, the EU legislation is containing a labelling obligation of 14 foods (2003/89/EC, 2006/142/EC), including cereals (gluten) as one of the eight major allergens. According to Codex regulation (Codex stan. 118-1979) foods containing less than 20 mg/kg gluten can be declared as gluten-free. To determine the gluten level of a product, application of reliable and validated analytical tools has utmost importance. However, proper validation of methods for allergen analysis has some limitations, e.g. lack of reference methods and materials. An additional problem is that effects of food processing on measurable allergenic protein content are not known perfectly. As a member of the Allergen Working Group within MoniQA Network of Excellence, the goal of our work is revealing the possible solutions of the above mentioned problems by development of an incurred reference material for the quantification of gluten. As a model matrix gluten-free flour based cookies were produced containing a dedicated amount of gliadin as the allergen source. With the help of this matrix, the effects of processing (heat treatment) could be studied as well. For this purpose an experimental design was made which contained the analytical investigation of every processing step: the mixture of the dry components, the raw dough and the baked products. Analysis of the gluten content was carried out by application of ELISA method. Results of the preliminary experiments indicated improper homogeneity of gliadin in the baked products and insufficient recovery values in the unprocessed mixture of dry components. These problems were eliminated by some changes in the applied workflow according to the results of gliadin solubility experiments and by the application of a newly developed homogenizing method. Another set of samples with proper homogeneity of gliadin were produced and with the help of these samples a comparative study of ELISA methods provided by different manufacturers has been started. With the help of the gathered data the effects of heat treatment could also be examined. First results show a measurable decrease of the gliadin content after heat treatment but this phenomenon needs further confirmation. These results are expected to be a contribution for proper method development and validation. Keywords: cereal allergy and intolerance, ELISA, method validation References Breitender H, Radauer C. Journal of Allergy and Clinical Immunology. 2004; 113 (5): 821-830 Hischenhuber C, Crevel R, Jarry B, Mki M, Moneret-Vautrin D A, Romano A, Troncone R, Ward R. Alimentary Pharmacology&Therapeutics. 2006; 23: 559-575 FDA Threshold Working Group (corr. author: Gendel S). Journal of Food Protecion. 2008; 71 (5): 1043-1088

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P11 H, C, N AND S STABLE ISOTOPE RATIOS AND ELEMENTAL DATA FOR THE TRACEABILITY OF ITALIAN PDO CHEESES
1 1

F. Camin 1 1 1 1 2 3 R. Larcher, D. Bertoldi, L. Bontempo, L. Ziller, G. Nicolini, M. Nocetti, A. Stroppa 1 IASMA Fondazione Edmund Mach, Via E. Mach 1, 38010 San Michele allAdige (TN), Italy 2 Consorzio Formaggio Parmigiano-Reggiano, via Kennedy 18, 42100 Reggio Emilia, Italy 3 Consorzio Tutela Grana Padano, Via XXIV Giugno 8, 25010 San Martino Della Battaglia Desenzano del Garda (BS), Italy

In this study we investigated the possibility of creating traceability models based on analytical data for several Italian PDO cheeses (Grana Padano, Parmigiano Reggiano, Asiago, Montasio, Toma, Fontina and Spressa). 50 authentic samples of Grana Padano, 76 samples of Parmigiano Reggiano and 96 samples from three potential commercial competitors, from Italy (37), the Czech republic (29) and Lithuania (30), as well as 79 alpine cheeses (16 Asiago, 16 Montasio, 16 Toma, 16 Fontina and 15 Spressa) were considered. As analytical markers we took into account: the stable isotope ratios of H, C, N and S analysed in the extracted casein of the cheeses using Isotopic Ratio Mass Spectrometers connected to an Elemental Analyser or a Pyroliser, the content of Ag, Al, As, Au, B, Ba, Be, Bi, Ca, Cd, Ce, Co, Cr, Cs, Cu, Dy, Er, Eu, Fe, Ga, Gd, Ge, Hg, Ho, Ir, K, La, Li, Mg, Mn, Mo, Na, Ni, P, Pb, Pd, Pr, Rb, Re, Sb, Se, Sm, Sn, Sr, Te, Tl, Tm, U, V, Y, Yb, Zn in cheese after microwave acid digestion, analysed using an Inductively Coupled PlasmaMass Spectrometer. The statistical models (Canonical Discriminant Analysis), structured on the basis of the most significant analytical parameters, were 100% effective in discriminating origin for Grana Padano and Parmigiano Reggiano and 94% for the alpine cheeses. The models can be proposed as suitable tool for verifying the authenticity of commercial samples and therefore for detection of mislabelling and for consumer protection. Keywords: IRMS, ICP-MS, traceability model

Poster Presentations

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P12 THE INFLUENCE OF POST-HARVEST STORAGE CONDITIONS UPON THE MICROBIOLOGICAL CONTAMINATION ON PEACH AND NECTARIN FRUITS A.Chira L. Chira, C.Alexe * University of Agricultural Sciences and Veterinary Medicine Bucharest - Romania * HORTING Research Institute Bucharest - Romania

ABSTRACT The most important link in the post-harvest technology is constituted by the physical treatment during the storage period. In the present paper we want to demonstrate the influence of the post-harvest storage conditions on the quality and shelf life of the peach and nectarine fruits. The experiment has been organized at the Faculty of Horticulture Science of Bucharest and at the HORTING Research Institute of Bucharest with four peach and nectarin varieties: Southland,Cardinal, Cora and Delta. The peach and nectarine fruits were harvested at the harvesting ripeness. At this moment, the main quality physico-chemical properties were analyzed, afterwards, the fruits were stored in different storage conditions, which represent the experimental variants: V1 = ambiental storage conditions : (T=20 C; RH= 68 %) V2 = refrigerating storage and microperforated plastic film: (T=2 C;RH=85%) V3 = refrigerating storage and semipermeable plastic film modified atmosphere: (T=2 C;RH=90%) The lower moisture percent and loss weight, for all varieties has been registered at variant were the fruits are stored under modified and refrigerated atmosphere. The depreciation due to rottenness and physiological disorders presented higher values in V1 case, while in V3 we can find the lowest values. Different varieties registered different reactions. Thus for Cardinal, the percentage of rotten fruits was of 13,5%, after a 5 days storage period (V1) in comparison with V 3 4 % after a 25 days storage period (V3). In Cardinal varieties case, the presence of the pathogen agents that caused fruits depreciation has been influenced by the storage as follows: the Monilinia laxa has developed better in low temperature conditions and high relative humidity (V2, V3, ) in comparison with V1. Botrytis cinerea has manifested itself stronger at a high temperature (V1). The physiological disorder (such as interval decay and fruit dehydration) was more significant in V1 (9%) as compared to V2 (5 %), and V3 (4 %). KEYWORDS: post-harvest, modified atmosphere, rottenness REFERENCES: Chira A., Chira L., Delian E., Stoian E., Roman M. Research concerning the influence of the pre and post-harvest phytosanitary treatments on peach and nectarine fruits quality. ESNA Amiens Meeting, 29 august 2 sept. 2005, France. Cuquel F. L., Monte B. Quality of peach fruits under integrated fruit production management.Acta Hort. 713, ISHS 2006.

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P13 COMPARISON OF ACRYLAMIDE CONTENT IN BREADS DIFFERING IN FORMULAS Z. Ciesarova 1 1 1 1,2 2 3 3 K. Kukurova , J. Sadecka , P. Simko , L. Markova , R. Belkova , T. Behan , J. Kravec 1 VUP Food Research Institute, Bratislava, Slovak Republic 2 VUT University of Technology, Faculty of Chemistry, Brno, Czech Republic 3 Mspomix, s.r.o., Zvolen, Slovak Republic
1

ABSTRACT: The occurrence of acrylamide in baked cereal products is considered to be the substantial source of acrylamide intake in human diet even due to a widespread consumption of various cereal products. One of the ways how to reduce this probably carcinogen exposure is to eliminate acrylamide level in the most frequently consumed items, which is bread. Since the acrylamide formation strongly depends on bread composition, the comparison of different bread mixes can reveal the connection between bread formula and following acrylamide content. In this study the acrylamide was determined in breads prepared from different bread mixes. The influence of different bread composition and the presence of some bread additives such as sodium phosphate, sodium pyrophosphate, sodium acetate, calcium chloride, calcium sulphate, citric acid, ascorbic acid, guar gum, glucono-delta-lacton etc. were evaluated. The goal of this study is to improve bread formula in such way which leads to acrylamide formation in smaller extent, but expected sensory properties are maintained. KEY WORDS: acrylamide, bread, additives ACKNOWLEDGEMENT: This contribution is the result of the project implementation "The Centre of Excellence for Contaminants and Microorganisms in Foods" supported by the Research & Development Operational Programme funded by the ERDF. This work was also supported by the o Slovak Research and Development Agency under the contracts N . LPP 0310-09, SK-PL-0051-09, and VMSP-P-0089-09.

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P14 METHOD DEVELOPMENT FOR THE DETECTION OF POTENTIALLY ALLERGENIC FINING AGENTS IN WHITE WINE: LYSOZYME AND OVALBUMIN M. Corrales H. Chassaigne European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg 111, B-2440 Geel, Belgium E-mail: margarita.corrales@ec.europa.eu During the winemaking process potential allergenic proteins are traditionally used as processing aids. Milk caseins and egg white proteins are used during fining to remove phenolic and tannin compounds to yield clear wines. Additionally, lysozyme can be used as a food preservative to inhibit malolactic fermentation. Part of these proteins precipitate and are removed by filtration during the manufacturing process. However, some protein traces may still remain in the wine (EFSA 2004, 2007). Directive 2000/13/EC, which has been amended by Directives 2003/89/EC and 2007/68/EC, regulates food labelling. It lists allergenic foods whose presence has to be declared on food labels. Directive 2007/68/EC is based upon EFSA opinions adopted in 2007, which in turn are based on scientific data supplied by wine producers that showed that allergic reactions can occur with fined wines. The European Commission has agreed to extend the deadline before mandatory labelling of egg and milk proteins used as fining agents in wines until late 2010 in order to facilitate the smooth transition towards new wine labelling requirements. The development of accurate and sensitive methodology for detection of allergenic proteins in wines is crucial to enforce future legislation and to ensure the correct labelling of products. In this work, the suitability of a sample preparation method to analyse wine matrices spiked with fining agents was investigated by LC-MS. Recovery experiments were performed spiking a wine model matrix and white wine with lyzozyme and ovalbumin at a concentration of 0.2 mg protein/kg matrix, a level below which allergic responses tend to be relatively weak (Kirchner et al. 2009). Protein quantification and SDS-PAGE were used to verify the extraction efficiency. The stability of lysozyme and ovalbumin in a wine matrix and a model wine matrix is reported in this work. Keywords: Wine, fining agents, processing aids, sample preparation, lysozyme, ovalbumin. Literature:
Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies on a request from the Commission related to a notification from the Winemakers' Federation of Australia (WFA) on milk products, egg products and fish products used in the manufacture of wine pursuant to Article 6 paragraph 11 of Directive 2000/13/EC. EFSA J. (2004), 134: p.1-6 Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies on a request from the Commission related to a notification from Winemakers' Federation of Australia (WFA) and Australian Wine Research Institute (AWRI) on casein and potassium caseinate used in the manufacture of wine pursuant to Article 6 paragraph 11 of Directive 2000/13/EC for permanent exemption from labelling. EFSA J. (2007), 531: p.1-6. Directive 2000/13/EC. European Parliament and Council, Off. J. Eur. Commun. (2000), L109: p. 29-42. Directive 2003/89/EC. European Parliament and Council.Off J. Eur. Commun. (2003), L308: p. 15-18. Directive 2007/68/EC. European Parliament and Council. Off J. Eur. Commun. (2007), L310: p.11-14. Kirchner, S., Belloni, B., Kugler, C., Ring, J., Brockow. Allergenicity of Wine Containing Processing aids: A double-blind, placebo-controlled food challenge. J. Investig. Allergol Clin, Immunol. (2009), 3: p. 210-217.

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P15 EFFECT OF BAKING CONDITIONS ON CRUST COLORATION AND ACRYLAMIDE FORMATION DURING BAKING OF WHITE BREAD T. Dessev *
( )

A. Le-Bail, J. Keramat(**), C. Prost, V. Lalane and V. Jury ONIRIS, UMR, GEPEA, CNRS 6144, BP 82225, F-44322 NANTES Cedex 3 France *University of Food Technologies, 26 Maritza blvd, Plovdiv 4002 Bulgaria **Department of Food Science and Technology, Isfahan University of Technology, Isfahan, 84156 - I.R.Iran

Abstract The effect of baking temperature (heart and vault) and steam injection on crust coloration and acrylamide formation during the baking of white bread was studied using response surface methodology with a central composite design. Baking was performed in a batch electrical deck oven (MIWE CO 1.1208 Germany) equipped with a stone hearth. The baking sets were done at different vault/hearth temperatures and steam injection levels in accordance with the experimental design matrix under natural convection. The steam injection was applied immediately after the start of baking. Response variables measured were crust coloration and acrylamide concentration. The CIE L*a*b (CIELAB) color model was chosen to describe browning development. The response was expressed as the total color difference. After baking the whole crust layer was carefully separated and dried at room temperature by oven with air circulation over night. Acrylamide was analyzed by gas chromatography mass spectrometry (GC-MC). The mass spectrometer was operated in selected ion 13 monitoring mode (SIM). Two ions were used to characterize C3-acrylamide (m/z 58 and 74) and two ions were used to characterize acrylamide (m/z 55 and 71). The ion m/z 71 was used to quantify acrylamide. Of the three process parameters, vault temperature exhibited the greatest effect (positive correlation) on crust coloration. As the vault temperature increased the crust coloration (total color difference) increased, too. Vault temperature and steaming were found to have greatest effect on acrylamide concentration. As they were increased the acrylamide concentration increased, as well. Additionally, a strong correlation between crust color and acrylamide concentration was observed. As the crust total color difference increased up to 20-25, the acrylamide concentration increased linearly. However for a higher level of crust coloration the acrylamide concentration was found to retain or even to decrease slowly. Keywords: acrylamide, bread, baking, crust
References: Ahrne L., Andersson C., Floberg P., Rosen J., Lingnert H. (2007). Effect of crust temperature and water content on acrylamide formation during baking of white bread: Steam and falling temperature baling. LWT 40, 1708-1715 Bagdonaite, K., & Murkovic, M. (2004). Factors affecting the formation of acrylamide in coffee. Czech Journal of Food Sciences, 22, 2224 Balaji, N. (1991). Modelling of transient temperature distribution during bread baking by finite difference analysis. B.Tech Thesis, IIT, Kharagpur, India Becalski, A., Lau, B. P. Y., Lewis, D., & Seaman, S. W. (2003). Acrylamide in foods: occurrence, sources, and modeling. Journal of Agricultural and Food Chemistry, 51(3), 802 808 Brathen E, Knutsen S, (2005). Effect of temperature and time on the formation of acrylamide in starch-based and cereal model systems, flat breads and bread. Food Chemistry 92, 693-700 Chul Soo Park and Byung-Kee Baik, (2007). Influence of baking and thawing conditions on quality of par-baked French bread. Cereal Chemistry 84(1), 38-43 Demirekler, P., Sumnu, G., Sahin, S. (2004). Optimization of bread baking in a halogen lamp-microwave combination oven by response surface methodology. European Food Research Technology. 219, 341-347 Eliasson, A.C., Larsson, K.(1993). Cereals in breadmaking. A molecular colloidal approach. New York: Marcel Dekker Inc. Grob, K., Biedermann, M., Biedermann-Brem, S., Noti, A., Imhof, D., Amrein, T., Pfefferle, A., & Bazzocco, D. (2003). French fries with less than 100 lg/kg acrylamide. A collaboration between cooks and analysts. European Food Research and Technology, 217(3), 185194

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Haase, N. U., Matthaus, B., & Vosmann, K. (2003). Acrylamide formation in foodstuffs minimising strategies for potato crisps. Deutsche Lebensmittel-Rundschau, 99 (3), 87-90 Jung, M. Y., Choi, D. S., & Ju, J. W. (2003). A novel technique for limitation of acrylamide formation in fried and baked corn chips and in french fries. Journal of Food Science, 68(4), 12871290 Krist-Spit, C.E., and Sluimer, P.(1987) Heat transfer in ovens during the baking of bread. In: Morton, I.D. (Ed), Cereals in a European Context. First European Conference on Food Science and Technology, New York: VCH, 344-354 Mottram, D. S., Wedzicha, B. L., & Dodson, A. T. (2002). Acrylamide is formed in the Mailard reaction. Nature, 419, 448449 Myers, R. H., Montgomery, D.C. (2002). Response surface Methodology (RSM): Process and product optimization using designed experiments. Wiley-Interscience Publication, USA, 1-3 Ozdemir, M., Devres, O. (2000). Analysis of color development during roasting of hazelnuts using response surface methodology. Journal of Food Engineering. 45, 17-24 Pyler, E.J. (1988). Baking science & Technology, vol.II (3 rd ed.). Meriam: Sosland Publishing Company Rydberg, P., Eriksson, S., Tareke, E., Karlsson, P., Ehrenberg, L., &Tornqvist, M. (2003). Investigations of factors that influence the acrylamide content of heated foodstuffs. Journal of Agricultural and Food Chemistry, 51(24), 70127018 Sablani, S.S., Marcotte, M., Baik, O.D., Castaigne, F. (1998). Modeling of simultaneous heat and water transport in the baking process. Lebensmittel-Wissenschaft und-Technologie 31(3), 201-209 Scanlon, M.G., Zghal, M.C. (2001). Bread properties and crumb structure. Food Research International, 34(10), 841-864 Sevimli, K. M., Sumnu, G., Sahin, S. (2005). Optimization of halogen lamp-microwave combination baking of cakes: a response surface methodology study. European Food Research and Technology. Shibukawa S., K. Sugiyama and T. Yano, (1989). Effects of heat transfer by radiation and convection on browning of cookies at baking, Journal of Food Science, 54, pp. 621624 699 Sommier, A., Chiron, H., Colonna, P., Della Valle, G., Rouille, J. (2004). An instrumented pilot scale oven for the study of French bread baking. Journal of Food Engineering, 69, 97-106 Stadler, R. H., Blank, I., Varga, N., Robert, F., Hau, J., Guy, P. A., Robert, M. C., & Riediker, S. (2002). Acrylamide from Maillard reaction products. Nature, 419(6906), 449450 Surdyk, N., Rosen, J., Andersson, R., & Aman, P. (2004). Effects of asparagine, fructose, and baking conditions on acrylamide content in yeast-leavened wheat bread. Journal of Agricultural and Food Chemistry, 52(7), 2047 2051 Tareke, E., Rydberg, P., Karlsson, P., Eriksson, S., & Trnqvist, M.(2002). Analysis of acrylamide, a carcinogen formed in heated foodstuffs. Journal of Agricultural and Food Chemistry, 50(17),4998 5006 Therdthai, N., Zhou, W., Adamczak, T. (2002). Optimisation of the temperature profile in bread baking. Journal of Food Engineering, 55(1), 41-48 Thorvaldsson, K., Janestad, H. (1999). A model for simultaneous heat, water and vapour diffusion. Journal of Food Engineering, 40, 167-172 Wahlby, U., Skjoldebrand C. (2002). Reheating characteristics of crust formed on buns, and crust formation. Journal of Food Engineering 53 (2), 177-184 Weisshaar, R., & Gutsche, B. (2002). Formation of acrylamide in heated potato products model experiments pointing to asparagines as precursor. Deutsche Lebensmittel-Rundschau, 98(11), 397400 Yam, K. L., Papadakis, S. E. (2004). A simple digital imaging method for measuring and analyzing color of food surfaces. Journal of Food Engineering, 61, 137-142 Zanoni, B., Peri, C., Bruno, D. (1995). Modeling of starch gelatinization kinetics of bread crumb during baking. Lebensmittel-Wissenschaft und Technologie, 28, 314-318 Zhang, J., Datta, A.K. (2006). Mathematical modeling of bread baking process. Journal of Food Engineering 75 (1), 78-89 Zhang, L., Lucas, T., Doursat, C., Flick, D., Wagner, M. (2007). Effect of crust constraints on bread expansion and CO2 release. Journal of Food Engineering , 80, 1302-1311 Zyzak, D. V., Sanders, R. A., Stojanovic, M., Tallmadge, D. H., Eberhart, B. L., Ewald, D. K., et al. (2003). Acrylamide formation mechanism in heated foods. Journal of Agricultural and Food Chemistry, 51(16), 4782 4787.

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Poster Presentations

P16 UNIVERSAL BIOLOGICAL BAR-CODE FOR DETERMINING THE GEOGRAPHICAL ORIGIN OF FRUITS BY USING PCR-DGGE A. F. El Sheikha * 1 D. Montet * Corresponding author Tel.: 33 4 67 61 57 28 Fax: 33 4 67 61 44 44 E-mail: elsheikha_aly@yahoo.com 1 Minufiya University, Faculty of Agriculture, Department of Food Science and Technology, 32511 Shibin El Kom, Minufiya Government, Egypt 2 Centre de Coopration Internationale en Recherche Agronomique pour le Dveloppement, CIRAD, UMR Qualisud, TA 95B/16, 34398 Montpellier Cedex 5, France
1, 2,

Introduction: The determination of geographical origin is a demand of the traceability system of import-export foodstuff. One hypothesis of tracing the source of a product is by analyzing in a global way the microbial communities of the food and links statistically this analysis to the geographical origin of the food (El sheikha et al., 2009a). Physalis is included in the priority list of many governments horticulture and fruit export plan. It is exported from several countries including Colombia, Egypt, Zimbabwe and South Africa, but Colombia stands out as one of the largest producers, consumers and exporters. Colombia exports of Physalis in 2004 were worth 14 millions USD (El Sheikha et al., 2008). In Egypt, economical importance of Physalis is rising, due to, achieving a great success in local, Arabic and European markets (El Sheikha, 2004). Physalis as the whole plant has many medicinal properties, including antipyretic, depurative, diuretic, pectoral, and vermifuge. A decoction is used in the treatment of abscesses, cough, fevers or sore throat (Duke and Ayensu 1985). The pulp is nutritious, containing particularly high levels of carotenoids, minerals, essential amino acids and vitamin C (El Sheikha et al., 2009b). Regarding Shea tree fruits, only seven countries have statistics. Nigeria accounts for more than 60% of the production of shea butter in 2005. It is followed by Mali, Ghana and Burkina Faso, which together account for just under a third of world production in 2005. In Europe, Shea butter is used mainly (95%) by the chocolate industry (FAOSTAT, 2007). The quantities exported to Japan, the United States or Switzerland would be mainly used for cosmetic or pharmacological (UNCTAD, 2001). Purpose: We applied a molecular technique employing 28S rDNA profiles generated by PCR-DGGE as a new traceability technique to detect the variation in fungal community structures of Physalis fruits from four countries (Colombia, Uganda, Egypt, Madagascar) and Shea tree from four countries (Cameroon, Mali, Senegal, Ghana). Results: The DGGE gels showed some significant differences in the migration patterns. However, the duplicates for each sampling location gave statistically similar DGGE patterns throughout the study. We demonstrated that there was a link between the fungi populations and the geographical area. When the 28S rDNA profiles were analyzed by multivariate analysis, distinct microbial communities were detected. The band profiles from different countries were different and were specific for each country and could be used as a bar code to discriminate the origin of the fruits. Significance of study: This method is a new traceability tool which provides fruit with a unique bar code and makes it possible to trace back the fruits to their original country. Key words: biological bar-code, fruits, PCR-DGGE

References

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Duke, J.A., and Ayensu, E.S.1985. Medicinal Plants of China Reference Publications, Inc., Algonac, Michigan, USA, 705 pp. El Sheikha, A.F. 2004. Technological, chemical and microbiological studies on some packed foods. MSc Thesis, Faculty of Agriculture, Minufiya University, Egypt, 174 pp. El Sheikha, A.F., Zaki, M., Bakr, A., El Habashy, M., and Montet, D. 2008. Physico-chemical properties and biochemical composition of Physalis ( Physalis pubescens L.) fruits. Global Science Books Ltd., UK, FOOD 2 (2) 124-130. El Sheikha, A.F., Condur, A., Mtayer, I., Le Nguyen, D.D, Loiseau, G., and Montet, D. 2009a. Determination of fruit origin by using 26S rDNA fingerprinting of yeast communities by PCR-DGGE: Preliminary application to Physalis fruits from Egypt. Yeast 26 (10): 567-573. El Sheikha, A.F., Zaki, M., Bakr, A., El Habashy, M., and Montet, D. 2009b. Biochemical and sensory quality of Physalis (Physalis pubescens L.) juice. Willy-Blackwell Publishing, Journal of Food Processing and Preservation (Early View). DOI: 10.1111/j.1745-4549.2009.00382.x. Available at: http://www3.interscience.wiley.com/cgi-bin/fulltext/123203771/PDFSTART Food and Agriculture Organization of the United Nations Statistics FAOSTAT. 2007. Shea Tree Fruits Production and Distribution in all of the World. UNCTAD. 2001. United Nations Conference on Trade and Development. Organ of the UN. UNCTAD secretariat based on the statistics of the United Nations Food and Agriculture. Market information in the field of commodities Shea Tree Fruits, N 2885. Available at: http://unctad.org/infocomm/francais/karite/plan.htm. Accessed February 2001.

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P17 THE DISTRIBUTION OF FUNGI AND MYCOTOXINS IN SOUTH AFRICAN MAIZE MILLS C. Erasmus (b). G.J. Marais (a) a. Centre for Applied Mycological Studies, Forestry and Biotechnology Institute, University of Pretoria, Lunnon Road, Pretoria, 0002, South Africa b. Agroprocessing and Chemical Technologies, CSIR Biosciences, Meiring Naud Road, P.O. Box 385, Pretoria, 0001, South Africa. e-mail: cerasmus@csir.co.za Fungi and their mycotoxins are regularly associated with maize and maize products throughout the world. However, the exact nature and effect of processing conditions and systems on the distribution (quantity) and type (species) of fungi and their associated mycotoxins as distributed through the South African maize milling system is poorly understood. Milled white maize products are the staple food of South Africans, especially amongst the rural poor. Products are consumed as a gruel or porridge or as a fermented drink and are eaten as a major part of all meals. This indicates a critical need to understand the risk associated with potential exposure to mycotoxins because of the high amounts of maize being consumed in relation to other foods. A situational analysis was done by comparing a selection of commercial mills in South Africa in terms of current fungal populations and mycotoxin levels. Samplings were done from all product streams and processing points in mills, and analysed using fungal enumerations (up to species levels), and mycotoxins (using the VICAM system and LC-MS/MS). A statistical approach was designed to determine the presence or absence of both field and storage fungi, and were analyzed separately. The observations were reported as percentages of presence and absence of a specific fungus and in the results these percentages were used as weights. Results indicated that fungi such as Fusarium (Section: Liseola), Cladosporium cladosporioides, Aspergillus flavus, Aspergillus clavatus, Penicillium spp., Mucor spp., and Rhizopus oryzae are the most dominant fungi in the maize milling system in South Africa. In all mills, irrespective of location, a constant level of fumonisins was present. These were identified as originating from the fields as the levels were the same as for whole maize at intake. In all mills, regardless of location, Aspergillus, Penicillium, Rhizopus and Mucor spp. were very low or nonexistent at intake, but increased significantly (P<0.05) after the conditioning steps during processing. These fungi were associated with the bran (chop) fractions and occurred in high numbers in end products derived from the bran fractions while levels in products derived from the endosperm (for example maize grits) were much lower. This indicated the occurrence of fungal germination during the conditioning steps and showed that the milling system can potentially modify the population of fungi found in South African maize by favouring the growth of storage fungi. No aflatoxins were detected in any samples while fumonisins were already present at intake showing that mills may not have a significant influence on mycotoxin levels naturally occurring in these products. KEYWORDS: Maize Mills, staple food, fungal populations, mycotoxins, South Africa
REFERENCES: Erasmus, C & Taylor, JRN, 2004. Optimizing the determination of maize endosperm vitreousness by a rapid nondestructive image analysis technique. Journal of the Science of Food and Agriculture, 84(9), 920 930. Galaverna, G., C. Dall'Asta, M. Mangia, A. Dossena & R. Marchelli (2009). Masked mycotoxins:an emerging issue for food safety. Csech Journal of Food Science 27: S89-S92. Gelderblom, W.C.A., N.P.J. Kriek & W.F.O Marasas. (1991) Toxicity and carcinogenicity of theFusarium moniliforme metabolite, fumonisin B1, in rats. Carcinogenesis 12: 1247-1251. Hoseney, R.C. (1994) Dry milling of cereals. In: Principles of Cereal Science and Technology. 2nded. Ed: Hoseney, R.C., St Paul: American Association of Cereal Chemists, Inc. pp125-145 Purchase, I.F.H., M. Steyn & H.E. Pretorius. (1967) The production of aflatoxin M on various substrates. Mycopathologia et Mycologia Applicata 24 (7): 239-244. Rabie, C.J., A. Lubben, G.J. Marais & H. Jansen van Vuuren. (1997) Enumeration of fungi in barley. International Journal of Food Microbiology 35: 117-127. Rabie, C.J. & G.J. Marais. (2000) Toxigenic fungi and mycotoxins in South African foods and feeds. Report to the Department of Health, 31 March 2000, 113pp.

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P18 VALIDATION OF ANALYSIS METHOD OF Cu METAL ON CANNED FOOD WITH AAS (ATOMIC ABSORPTION SPECTROSCOPY) INSTRUMENT Faridah, D.N 2) D.Herawati PhD Student, Food Science Food Analysis Laboratory, Department Food Science and Technology, Bogor Agricultural University (IPB), Indonesia
1)

The aim of the experiment is to study the validation method of Cu in food using AAS. The validation of method used to canned food different with the standard matrix method (AOAC 975.03 and 965.09 for plant matrix). The parameter used to validate the performance of AAS instrument were done using a determination of linearity of calibration curve, sensitivity and the value of LoD ( Limit of Detection) with Cu standard (7 concentration different with 7 replicate). The validation of analysis method was done using determination of LoQ (Limit of Quantification), precision and accuracy. The linearity of the 2 calibration curve can be seen from the R value with 0.9997 and sensitivity of the calibration curve was slope with value of 0.1085. Slope data and calibration curve intercept can be used as a quality assurance in the next analysis of Cu, the LoD was 5 ppb (10 independent replicate, 3 times of SD/Standard Deviation blank). The LoQ was 10 times SD (10 independent replicate) with value of 50 ppb. Precision can be seen from the RSD (Relative Standard Deviation) value and was done with standard spike of Cu in the sample with concentration of 2, 5 and 20 ppm (7 replicate). The precision was quite good since the RSD value was lower than the RSD Horwitz with recovery 87.31-96.58%. The validation method is important to justify if differences exist using the standard method. Key words: AAS, method validation, RSD REFERENCES AOAC. 2002. Appendix D. Guidelines for Collaborative Study Procedures to Validate Characteristic of A Method of Analysis. Eurachem. 1998. The Fitness for Purpose of Analytical Methods. First English Edition 1.0. Frederick M. Garfield, Eugine Klesta, Jerry Hirsch. 2000. Quality Assurance Principles for Analytical Laboratories. Third Edition. AOAC International. Gaithersburg, MD USA. Nielsen Suzanne. 2003. Food Analysis. Third Edition. Kluwer Academic/Plenum Publishers. New York.

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P19 A NOVEL HIGH-RESOLUTION MASS SPECTROMETRY STRATEGY FOR THE NON-TARGETED DETECTION OF CHEMICALS IN FOOD M. Godula 2 2 A. Charlton , D. Roberts 1 Thermo Fisher Scientific, Slunen 27 Praha 10, 100 00, Czech Republic michal.godula@thermofisher.com 2 Food and Environmental Research Agency, Sand Hutton, York YO41 1LZ, UK
1

There is a great need for analytical methodologies for holistic monitoring of food, an approach often referred to as profiling or screening. Currently, chemical analysis of food is largely limited to the direct determination of compounds that are specified prior to analysis, the so-called target list approach. Whilst this approach is highly relevant to the enforcement of regulatory parameters relating to known food contamination issues (such as the detection of pesticides and veterinary drug metabolites in foods) it does little to address those components in food that are not anticipated. A number of high profile issues have highlighted the need for broader ranging determination of the composition of foodstuffs and most recently the fatalities that arose from the contamination of milk with melamine have underlined this. By obtaining a detailed understanding of what is naturally and normally present in foods it becomes possible to monitor the food chain for abnormalities. A profiling or screening approach thus facilitates the determination of a wide range of issues in the areas of food fraud and food safety. Whilst holistic monitoring of food has been the holy grail for a number of years, it is only in recent times that instrumentation technology has advanced to the point where this has become feasible. The poster presented will show the development and validation of simple analytical approach using high resolution mass spectrometry as a tool for rapid analysis of food samples for a variety of contaminants. The methodology presented was validated using milk powder spiked with a diverse range of contaminants, including melamine to demonstrate the feasibility of simultaneously detecting compounds for which different targeted approaches would normally be used. The main benefits of such an approach are relatively easy sample preparation strategy and short analysis times. Very high resolving power (up to 100.000 FWHM) delivered by the new single stage mass spectrometer TM TM Exactive based on the proved Orbitrap technology allows to analyze variety of contaminants in relatively dirty extracts at very low concentration levels. The high resolution applied allows better separation of interfering compounds from analytes of interests and thus leads to excellent mass accuracy values necessary for identification of analytes. To uncover the presence of contaminants in the matrix the comparative software Sieve was used. Sieve employs specific data processing algorithms enabling identification of differences between data sets (blank and contaminated) and their automated identification via web based databases.

Keywords: non-target screening, high resolution MS, food profiling

Poster Presentations

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P20 EFFECT OF TRANSGLUTAMINASE ENZYME AND MAILLARD REACTION ON THE LOSS OF LYSINE IN CRACKERS Seher Gm 1 1 V. Gkmen , H. Kksel 1 Food Engineering Department, Hacettepe University Ankara, TURKEY
1

Transglutaminase is a cross-linking enzyme catalyzing an acyl-transfer reaction between the carboxyamide group of peptide-bound glutamine residues (acyl donors) and a variety of primary amines, including the -amino group of lysine residues to form an -( -glutamyl) lysine bond. In earlier stages of Maillard reaction, a reducing sugar, like glucose, condenses with a compound possessing a free amino group (e.g. amino group of amino acids, -amino group of lysine and amino group of N-terminal amino acid in proteins). The resulting condensation product, N-substituted glycosylamine, rearranges to form the Amadori product. Furosine is a marker of the degradation of lysine residues in protein and is formed during acid hydrolysis of the Amadori compound generated at the early stage of the Maillard reaction in thermally treated foods. The loss of available lysine is the most significant consequence of this reaction, and it is of great importance in those foods where this amino acid is limiting, such as in cereals as well as other processed foods (Erbersdobler and Hupe, 1991). In this study, the influence of cross-linking by transglutaminase enzyme between glutamine and lysine on the Maillard reaction is investigated. For this purpose, the effect of the different time/ temperature parameters and different levels of enzyme are examined by measuring color values (L*, a*, b*) and furosine contents of the samples. The color change was observed between control samples and the samples supplemented with transglutaminase enzyme. The decreases in color values were obvious in enzyme supplemented samples. There were also the differences between the color values of the samples supplemented with different levels of enzyme. The chemical analyses are being carried out to support the color analysis results. KEYWORDS: Transglutaminase, Maillard reactions, Lysine loss, Furosine REFERENCES: Basman A., Kksel H., Ng P. K. W. 2002. Effects of increasing levels of transglutaminase on the rheological properties and bread quality characteristics of two wheat flours. Euro Food Research Technology. 215:419424 Gan C.-Y., Cheng L.-H., Easa A.M. 2009. Assessment of Cross-Linking in Combined Cross-Linked Soy Protein Isolate Gels by Microbial Transglutaminase and Maillard Reaction. Journal of Food Science. 74(2): C141-6 Gkmen V., Serpen A., Morales F.J. 2009. Determination of Furosine in Thermally Processed Foods by Hydrophilic Interaction Liquid Chromatography. Journal of AOAC International. 92(5):1460-3. Motoki M, Seguro K. 1998. Transglutaminase and its use for food processing. Trends Food Science Techology. 9: 20410. Ramirez-Jimenez A., Guerra-Hernandez E., Garcia-Villanova B. 2003. Evolution of non-enzymatic browning during storage of infant rice cereal. Food Chemistry. 83: 219225

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P21 ALLERGEN MANAGEMENT: TRACKING DOWN HIDDEN ALLERGENS WITH LATERAL FLOW RAPID TESTS M. Richter , S. Haas-Lauterbach , K. Schmitt 1 R-Biopharm AG, An der neuen Bergstrasse 17, 64297 Darmstadt, Germany, info@r-biopharm.de 2 bioavid Diagnostics GmbH & Co. KG, Schlossgasse 17, 64807 Dieburg, Germany, info@bioavid.de Presented by: Christine M Gutschelhofer
1 1 2

Allergen management in food companies is becoming increasingly important. Health risks due to contaminated food must be detected and prevented in time (Taylor & Hefle 2005). A standardized protocol for clinical experiments that allows determination of the threshold dose is needed (Taylor, Hefle et al. 2002) (Taylor, Gendel et al. 2009). HACCP concepts are mandatory in food producing firms and are the best approach to control the risks in food products. The HACCP concept (article 5 EG hygiene directive 852/2004) requires checking harmful residues in food. However, efficiency of measurements has been difficult to monitor due to the lack of reliable methods for allergen detection that are safe and easy to be used on site. It is possible to carry out residue and hygiene controls by means of immunochemical tests. Techniques like ELISA (Enzyme-linked Immunosorbent Assay), lateral flow devices and PCR (Polymerase Chain Reaction) are available on the market. R-Biopharm is offering all three techniques in its product range for allergen management systems for many different parameters. Lateral flow technique in combination with swabbing tests are ideal tools for hygiene monitoring. The bioavid lateral flow rapid tests are designed for HACCP based hygiene monitoring to guarantee production of allergen free and safe food. The tests are presenting a new set of lateral flow devices that are unique in design, performance, and ease of use. The bioavid lateral flow product portfolio comprises analysis of Almond, Brazil Nut, Coconut, Mustard, Hazelnut, Macadamia Nut, Peanut, Egg, Sesame, Cashew Kernel, Pistachio and Milk. These tests can be applied to test food products, rinse water (CIP), and surfaces for contamination. These lateral flow allergen tests can hide allergens rapidly and reliably even while the production process is underway to follow the requested HACCP plan. Cross-contaminations become detectable and preventable. The sensitivity of the specific lateral flow tests is comparable to that of commercial ELISAs for the matrices under investigation. Lateral flow allergen tests contain specific antibodies which are conjugated to tracers and which are delivered in a dried and stable format. After activation and run of the specific antigen-antibody reaction, coloured lines appear on the test strip. The bioavid allergen tests are tools for efficient detection of allergen contamination in production lines, and in food. In particular the ease of use and the speed of analysis make them valuable for the implementation and monitoring of HACCP concepts in the food industry. References: Taylor S.L. & Hefle S.L. (2005) Allergen Control Increased awareness of food allergies is leading food companies to develop comprehensive allergen control programs. Heres whats involved. Food Technology; Vol.59, No. 2 Taylor, S.L., Hefle, S.L. et al. (2002) Factors affectiong the determination of threshold doses for allergenic foods: How much is too much? J Allergy Clin Immunol; Vol. 109, No.1 Taylor, S.L., Gendel, S.M., Houben, G.F. and Julien, E. (2009) Critical Review in Food Science and Nutrition Vol. 49

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P22 ESTABLISHMENT OF A LAB MADE MAGNETIC BEADS AND MULTIPLEX PCR SYSTEM FOR THE SIMULTANEOUS DETECTION OF ESCHERICHIA COLI O157:H7, SALMONELLA AND SHIGELLA Haojiang Zuo 1 1 2 1 Xiaobei Ding , Wei Huang , Xuejun Fan ,Jingyan Yang , Xiaofang Pei* 1 West China School of Public Health, Sichuan University, Chengdu, China; 2 Sichuan Entry-Exit Inspection and Quarantine Bureau
1

[Abstract] Escherichia coli O157:H7, Salmonella and Shigella are the major food borne pathogens. Simultaneous detection three of them will be very important for the prevention. Although multiplex PCR is a good choice, the number of the pathogens and inhabitants in the samples are the main influent factors for the detection limit of PCR. So we attempted to develop a cheap magnetic bead that can effectively capture several bacteria, and identify certain pathogens with the combination of multiplex PCR. Objective To study the adsorption capacity of lab made magnetic beads on the common food-borne pathogenic bacteria and establish a lab made magnetic beads and multiplex PCR system for simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella. Methods Different reaction condition (volume, time, temperature, pH) was conducted to optimize the bacteria adsorption capacity of lab made magnetic beads. The optimized beads were then used to adsorb different bacteria to evaluate their adsorption capacity. The PCR conditions were also optimized. The bacteria were captured non-specifically by magnetic beads from bacterial suspensions, and lysed by heating with a beads suspending enhancer; the supernatant was then assayed by multiplex PCR, using primers specifically targeting the uidA, invA and ipaH genes of Escherichia coli O157:H7, Salmonella and Shigella respectively. Then the optimized lab made magnetic beads and multiplex PCR system was applied to detect Escherichia coli O157:H7, Salmonella and Shigella in samples. Results Volume, time, temperature and pH are influential factors of the adsorption capacity of the beads. The lab made magnetic beads have a high effective adsorption capacity (more than 95%) on common food-borne pathogens and indicator bacteria at 37 in saline. The component of the tested samples could decrease the adsorption capacity of the magnetic beads (58.4%). The detection sensitivity varied among different samples. However, it was still better than that of traditional microbiological analyses. Nonspecific amplification product was not detected in PCR reaction when testing other 19 related species of bacteria strains. The whole procedure could be easily completed within 30 h whereas the traditional method need at least 3 days. Conclusion Magnetic separation technology can rapidly adsorb and condense bacteria. The lab made magnetic beads demonstrate a high adsorption capacity to the bacteria. The combined application of lab made magnetic beads and multiplex PCR offers a potentially convenient and rapid microbiological tool for simultaneous detection of E. coli O157:H7, Salmonella and Shigella in samples. [Key words] E coli O157:H7; Salmonella; Shigella; Magnetic bead; Multiplex PCR. [References] 1. Qiu Jin, Fan Xuejun, Shen Sheng, et al. Study on the adsorption capacity of lab made magnetic particles to the common food- borne pathogenic bacteria. Modern Preventive Medicine. 2006, 33(1): 4-5 2. Y. Li, S. Zhuang and A. Mustapha. Application of a multiplex PCR for the simultaneous detection of Escherichia coli O157:H7, Salmonella and Shigella in raw and ready-to-eat meat products. J Food Prot. 2004, 67(1): 27-33 ----------------------------* Corresponding author. Fax: +86 28 85501275 (fax); E-mail address: xxpeiscu@163.com This work was partially supported by MoniQA (FOOD-CT-2006-036337) and scientific research project of General Administratioin of Quality Supervision, Inspection and Quarantine of the Peoples Republic of China (NO. J2005J0115)

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P23 EXTRACTION OF WHEAT COMMODITIES WITH DIFFERENT EXTRACTION BUFFERS AND THEIR INFLUENCE ON SIGNAL INTENSITY IN TRIPLE-QUADRUPOLE MASS SPECTROMETRY J. Heick 2 3 M. Fischer , B. Ppping 1 Eurofins Analytik GmbH, Neulnder Kamp 1, 21079 Hamburg, Germany 2 Universitt Hamburg, Institut fr Biochemie und Lebensmittelchemie, Abteilung Lebensmittelchemie, Grindelallee 117, 20146 Hamburg, Germany 3 Eurofins Scientific Group, 69a Kilnwick Road, Pocklington, Yorks, YO42 2JY, UK
1, 2

Celiac disease is a chronic food-sensitive enteropathy induced by ingestion of storage proteins (gluten) from wheat, rye and barley. It is the most common food intolerance with approximately one in every 300 affected in Europe. Only treatment available is a strict gluten-free diet. In the European Union products containing less than 100 mg/kg gluten may be labelled with very low gluten, those with less than 20 mg/kg with gluten-free. Enzyme-linked immunosorbant assays (ELISA) are currently the most widespread methods to detect these amounts of gluten. Mass spectrometry has become the leading technology in protein identification and offers new possibilities for gluten detection. Methods based on triple-quadrupole mass spectrometers may also be used for quantitative analysis. Briefly, these methods rely on tryptic digestion of the protein, followed by an identification of the generated peptides. Bottleneck of every gluten detection method is the extraction from food matrices. Therefore the extractability is often enhanced with denaturing additives. However, these may result in reduced signal intensity in the mass spectrometer. In this work, a gluten detection method based on triple-quadrupole mass spectrometry was developed. The method is based on selection of appropriate peptides for multiple reaction mode (MRM). This was used to detect gluten in a variety of wheat commodities. Different extraction buffers and their influence on signal intensity were evaluated. Keywords: gluten, mass spectrometry, multiple reaction mode, extraction buffer E. Garcia, M.LLorente, A. Hernando, R. Kieffer, H. Wieser, E. Mndez, Eur. J Gastroenterol Hepatol, 17 (2005) 529-539. A. Gibert, M. Espadaler, M.A. Canela, A. Snchez, C. Vaqu, M. Rafecas, Eur J Gastroenterol Hepatol, 18 (2006), 1187-1195. H. Wieser, Z Lebensm Unters Forsch A 207 (1998) 128-132.

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P24 INHIBITION OF ESCHERICHIA COLI BY THE LACTIC ACID BACTERIUM LACTOBACILLUS PLANTARUM B33 IN MUSTARD LEAVES FERMENTATION P.-H. Ho S. Chu-Ky, K.T.M. Tran, T.M.H. Pham, T.T.H. Nguyen, and T.M. Le* School of Biotechnology and Food Technology, Hanoi University of Technology 1 Dai Co Viet, Hanoi, Vietnam * Corresponding author: Phone: (+84-4) 3868 0119 Fax: (+84-4) 3868 2470 Email: lethanhmai@mail.hut.edu.vn

Abstract This study is to examine microbiological quality of Vietnamese traditional fermented foods at small scale and to study the ability to use isolated lactic acid bacteria as starter culture to improve product quality. Fermented green mustard leaves are a popular traditional food in Vietnam. This product is normally manufactured by spontaneous fermentation at a small scale, therefore, easily to be contaminated. A microbiological survey on 32 samples of fermented mustard leaves available in markets in Hanoi during the period of November 2008 May 2009 was conducted. Total microbial count, yeast and mold, coliforms and Escherichia coli, Clostridium perfringens, and Bacillus cereus was determined based on Vietnamese standard methods. Number of lactic acid bacteria was also enumerated using plate count method on DeMan-Rogosa-Sharpe agar medium (MRS). Preliminary result showed that none of test samples met the regulation of Vietnamese Minister of Health (46/2007/Q-BYT) on coliforms and E. coli. With the aim to improve the quality and to industrialize production of fermented vegetables, we studied the inhibition activity of Lactobacillus plantarum B33 toward E. coli. The lactic acid bacterium L. plantarum B33 was previously isolated from Vietnamese traditional fermented products and characterized by high capability of producing lactic acid. This bacterium was found to reduce E. coli by 2 -1 2-4 x 10 CFU mL in MRS medium. When the strain B33 was used as a starter culture for mustard leave fermentation, tested E. coli was totally suppressed after 36 h of fermentation. Sensory properties of product were also significantly improved. This study suggested that the strain L. plantarum B33 could be applied in vegetable fermentation as a starter culture to scale up production, stabilize product quality and ensure food safety. Keywords: fermented vegetable, inhibition, Lactobacillus, Escherichia coli, starter culture

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P25 MOLECULARLY IMPRINTED SOLID-PHASE EXTRACTION COMBINED WITH HIGH PERFORMANCE LIQUID CHROMATOGRAPHY FOR DETERMINATION OF FURAN IN INSTANT COFFEE Xue-Song Huang* Ning Peng (Department of Food Science and Engineering, Jinan University, Gunag Zhou 510632, China)

Furan is naturally occurring compounds found at very low levels in many heat treated foods and drinks. Furan, classified to be possibly carcinogenic to humans (Group 2B) by the International Agency for Research on Cancer (IARC), has been highly paid attention to its toxity. It is hard to be determined for its volatility (BP=31.36 ) and trace in the food. By now, the mainly detection methods of furan were solid phase micro-extractiongas chromatography mass spectrometry (SPME/GC/MS) and head space-gas chromatography-mass spectrometry (HP/ SPME /GC/MS). In order to detect the trace furan in foodmolecular imprinted polymer (MIP) has been synthesized and used as a sorbent to enrich the trace furan to a high level at which could be detected by high performance liquid chromatography (HPLC) easily, accurately and efficiently. The MIP was prepared following a non-covalent protocol with 2-methyl furan as dummy template, methyl acrylic acid (MAA) as functional monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker and acetonitrile as porogen at 60 for 24h. Then, the binding behavior of the furan on the MIP was evaluated by HPLC. The MIP was filled into a column and then the procedure was executed following MIP- solid-phase extraction (SPE) protocol. The sample solutions were applied to the MIP-SPE column after the MIP was conditioned completely. Furan bound by the MIP-SPE column was washed and eluted with the different conditions (solvents, pH, etc) and determined by HPLC at 210 nm and 28. At last, the MIP was applied to selectively extract furan from instant coffee (Nestle). The results showed that the MIP has porous structure and highly specific absorption for furan ( Imprinted factor= QMIP/QNIP=1.879). The MIP-SPE protocol was carried out well when the MIP-SPE column conditioned by methanol, washed by acetonitrile: water (1:9 V/V) and eluted by methanol: acetic acid (99:1 V/V). The results also showed the mean recoveries of MIP-SPE were 65.17% and the low limits of detection were 40g/L for furan by HPLC. The furan were determined as 421.7 g/kg (n=3) in the instant coffee by MIP-SPE- combined with HPLC method, which correspond with the results announced by European Food Safety Authority (EFSA). So MIP-SPE-HPLC method could be used for the determination of trace furan in food. The method is easily and efficiently. However, for the accurate results, more research need to be done to optimized the procedure of SPE. Key Words: Furan, 2-Methyl Furan, MIP-SPE, HPLC Reference: IARC. (1995). IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. 63, 3194 3407. Bononi, M., &Tateo, F. (2009). Determination of furan by headspace solid-phase micro extractiongas chromatographymass spectrometry in balsamic vinegars of Modena (Italy). Journal of Food Composition and Analysis , 22, 7982. Bianchi, F., Careri. M., Mangia, A., &Musci, M. (2006). Development and validation of a solid phase micro-extractiongas chromatographymass spectrometry method for the determination of furan in baby-food. Journal of Chromatography A, 1102, 268272. EFSA. (2009). Results on the monitoring of furan levels in food. EFSA Scientific Report, 304, 1-23.

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P26 PRIORITIES AND NEEDS FOR IMPLEMENTATION OF UPDATED HACCP SYSTEMS AND MODERN MONITORING TECHNOLOGIES AT THE BULGARIAN FOOD SECTOR M. Ivanova G. Ivanova, V. Gotcheva, A. Angelov Department of Biotechnology, University of Food Technologies, Plovdiv, Bulgaria

An international survey on the implementation of updated HACCP-based systems and advanced monitoring technologies throughout the food sector was conducted under project MoniQA. Bulgaria took part in the survey as well, reaching different branches of the countrys food production sector. The distributed questionnaire covered the implementation of food safety and quality management systems, analyses of various process- and product parameters and the methods used, the need of rapid analytical methods, and the application of information communication technology (ICT) systems. Altogether 50 Bulgarian food enterprises responded with completed questionnaires, 92% of them being with SME status. The majority of the companies (84%) had certified food safety management systems. Microbiological contaminants comprised the largest group analysed (90%), followed by heavy metals (28%) and foreign bodies (28%). Regarding the needs of respondents for rapid analytical methods, microbiological contaminants also rank highest (62%). Many companies expressed need of rapid methods for food allergens (17%) and mycotoxins (13%) as well. Half of the respondents performed their analyses in external laboratories. The implemented ICT-based management systems were mainly used for storage area management (63%), raw materials- (58%) and product traceability (56%). ICTs were also used to source information about existing suppliers and customers, local legislation and regulations. Analysis of the collected data shows that despite the wide implementation of food quality and safety management systems, analyses performed still do not cover all relevant parameters at appropriate levels, and development of rapid instrumental analyses is of major need for the food companies. Keywords: HACCP survey, ICT systems, rapid methods, food production, food safety References: Ba, M., Ersun, A. ., Kvan, G., 2006, Implementation of HACCP and prerequisite programs in food businesses in Turkey, Food Control, 17:118-123. Griffith, C., 2000, Food safety in catering establishments, In J. M. Farber and E. C. D. Todd (Eds.), Safe handling of foods, 235-256, New York: Marcel Dekker. Jevnik ., Hlebec V, Raspor P., 2008, Food safety knowledge and practices among food handlers in Slovenia, Food Control, 19:1107-1118 McSwane, D., Rue, N., Linton, R., 2003, Essentials of food safety and sanitation (3rd edition), New Jersey: Pearson Education, 169196 Walker, E., Jones, N., 2002, An assessment of the value of documenting food safety in small and less developed catering businesses, Food Control, 13 (45), 307314

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P27 THE ANALYSIS OF STATUS OF THE STANDARDS SYSTEM OF FOOD CONTACT PLASTIC MATERIALS IN CHINA Chen Jin-yao Yao Xiao-fen, Zhang Li-shi Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University, China
Background/objectives: Plastic materials have been commonly served as food packaging materials. There is no denying that plastic material holds certain advantages over paper, metal and glass, but it also poses certain hazards due to migration of low-molecular compounds. Recently, there has been a growing public concern regarding the safety of food contact plastic materials on a global scale. The objective of this study is to get a comprehensive recognition of the standards system of food contact plastic material in China, and to provide some references for food inspection authorities, relevant industries and enterprises. Methods: National standards and industrial standards of food contact plastic materials in China were collected, compared and analyzed, and they were compared with the relevant regulations and standards in U.S and EU. Through this process, some defects and challenges were put forward. Results: The standards system of food contact plastic materials in China was basically composed of four categories of standards, i.e. general standards which are applied to all packaging materials, manufacturing practice for food contact plastic materials, standards for certain products including hygienic standards and quality standards, and the standards for test and analytic methods. Some defects found in the system included that (1) several government agencies were involved in publishing and revising of the standards, and some of them lack of guidelines for risk assessment; (2) contradiction of certain hygienic requirements existed in multiple standards for a certain product; (3) the revising procedure was lagged behind the industrial development; (4) certain gaps concerning hygienic requirements existed between the Chinese standards and those of developed countries(e.g. absence of quality standards for monomers and hygienic standards for both printing ink and adhesives compared to Commission directive 2002/72/EC); (5) there was some inconsistency in the standards for test methods with products standards, especially fast detecting methods. Conclusions/suggestions: The current standards system is not consummated enough to meet the need of food safety inspection in China, and also not enough to international trade. It was suggested that subsequent work should be done to improve the efficacy of the standards, including (1) reviewing and combining the mixed and perplexing standards to develop a single integrated standards system that will entail mapping the food continuum from production to consumption; (2) promoting renew and revising frequency; (3) improving science-based work, especially establishing the guidelines for risk assessment of food contact plastic material, as well as monitoring and rapid alert system; (4) strengthening validation work of test methods; (5) paying more attention to communication with relevant industries and consumers. Key words: food contact materials, plastic, standards References: < the compilation of standards for packaging materialsspecially for food >, Chinese standards publishing house, November, 2009 Wang Jing, Liu Wen. (2009). Contrast and analysis of indexes in standards of packing containers and materials for food both in China and abroad. Food Science and Technology, 34(4), 226-229. Commission directive 2002/72/EC. Relating to Plastic Materials and Articles Intended to Come into Contact with Foodstuffs[S]. Official Journal. 82/711/EEC. Laying Down the Basic Rules Necessary for Testing Migration of the Constituents of Plastic Materials and Articles Intended to Come into Contact with Foodstuffs[S]. Official Journal. 2007/19/EC. Amending Council Directive 2002/72/EC Relating to Plastic Materials and Articles Intended to Come Into Contact with Foodstuffs [S]. Official Journal. 93/8/EEC. Amending Council Directive 82/711/EEC Laying Down the Basic Rules Necessary for Testing Migration of Constituents of Plastic Materials and Articles Intended Come Into Contact with Foodstuffs [S]. Official Journal.

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P28 ACRYLAMIDE CONTENT IN COOKIES ENRICHED BY FRUIT FIBRE K. Kukurov 1 2 2 2 Z. Ciesarov , Z. Kohajdov , J. Karoviov , M. Jursov 1 VP Food Research Institute, Priemyseln 4, 824 75 Bratislava, Slovak Republic 2 Slovak University of Technology, Institute of Biotechnology and Food Science, Department of Food Science and Technology, Radlinskho 9, 812 37 Bratislava, Slovak Republic
1

At the present, the human diet is deficient in fibre, which leads to numerous health complications. Fibre incorporation, in frequently consumed food such as bakery products, could help to overcome the fibre deficit. There are two reasons to add fibre to baked products: the increase of dietary fibre intake and the decrease of the caloric density of baked goods. Traditionally, fibre supplementation into the bakery products is focused on the use of milling by-products. There are many other sources of dietary fibre, such as fruits mainly apple, orange, lemon and grapefruit. The main advantage of dietary fibre derived from fruits comparing to other alternative sources, such as cereals, is its higher proportion of soluble dietary fibre. Dietary fibres are not only desirable for their nutritional properties, but also for their functional and technological properties and because of those they can also be used to upgrade agricultural products and by-products for use as food ingredients. Acrylamide is a chemical compound which is formed during heat processing of the wide range of starchy food products. According to the International Agency for Research on Cancer (IARC) acrylamide is classified as a probable human carcinogen of the group 2A and for that reason FAO/WHO recommends to minimize its content in food products. Although the levels of acrylamide in most of cereal products are relatively low, final dietary exposure from this type of foods is significant. It is known that ingredients used in cereal products such as dries fruits (e.g. prunes, pears) may contribute significantly to the final acrylamide content. In this study the addition of different types of fruit fibre (apple, orange, lemon and grapefruit) to cookies in level of 5, 10, 15 and 35 % (w/w) was tested. A reference sample without fruit fibre addition contained 363 g/kg acrylamide determined by GC-MS/NCI. Lemon fibre resulted in the highest acrylamide increase up to 77718 g/kg at 35 % of this fibre addition. On the other hand, apple fibre did not affected acrylamide content in such extent and increased it noticeably only in higher levels of fibre content up to acrylamide concentration of 586 g/kg at 15 % of apple fibre addition, and 1037 g/kg at 35% of apple fibre addition, respectively. These observations serve as a base for considering a risk/benefit approach to food supplementation. KEY WORDS: acrylamide, cookies, fruit fibre ACKNOWLEDGEMENT. This contribution is the result of the project implementation "The Centre of Excellence for Contaminants and Microorganisms in Foods" supported by the Research & Development Operational Programme funded by the ERDF. This work was also supported by the Slovak Research and Development Agency under the contracts No. LPP 0310-09, SK-PL-0051-09, and VMSP-P-0089-09 and grant agency of the Ministry of Education of the Slovak Republic and of Slovak Academy of Sciences VEGA Grant No. 1/0570/08.

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P29 CHARACTERISATION AND CLASSIFICATION OF SERBIAN HONEY BY PHYSICOCHEMICAL PARAMETERS K. Lazarevi 1 2 2 2 M. Joveti , D. Milojkovi-Opsenica , F. Andri , . Tei 1 Center for Food Analysis, Zmaja od Noaja 11, 11000 Belgrade, Serbia 2 Faculty of Chemistry, University of Belgrade, Studentki trg 12-16, P.O.Box 51, 11158 Belgrade, Serbia
1

The characterization of four different monofloral and multifloral types of Serbian honey (a total of 350 samples collected from various regions of Serbia from 2009 harvesting season) was carried out on the basis of common physico-chemical parameters (moisture, ash, electrical conductivity, acidity, and optical rotation), following The Harmonised Methods of the International Honey Commission The analyzed samples were characterized and distinguished using several pattern recognition chemometrical methods. Cluster Analysis and Principal Component Analysis were used as a tool to highlight the data structure nding relationships between investigated physico-chemical parameters and botanical origin of honey. Key words: honey, physicochemical parameters, botanical origin, chemometrics References: Harmonised Methods of the International Honey Commission (IHC), International Honey Commission (2002). Revised Codex Standard for Honey, Codex STAN 121981, Rev. 1 (1987), Rev. 2 (2001), Codex Alimentarius Commission (2001). Bogdanov, S., Ruo, K., & Oddo, L. P. (2004). Physico-chemical methods for the characterization of unioral honeys: A review. Apidologie, 35,417. G.Saric et al., Characterisation of Croatian Honey, Food Technol. Biotechnol. 46 (4) 355 367 (2008)

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P30 IMPLEMENTATION OF HACCP SYSTEMS AND RAPID METHODS IN THE FOOD INDUSTRIES. D. Lebesi C. Dimakou, V. Oreopoulou & C. Tzia 1 Laboratory of Food Chemistry and Technology, School of Chemical Engineering, National Technical University of Athens, 5 Iroon Polytechniou St., 15780 Athens, Greece

HACCP is a driving force for the development of validated rapid methods. Successful combination of a HACCP system and rapid methods may help the industry to find new ways of obtaining reliable results. Rapid methods are among the more valuable tools available to companies who are considering implementing HACCP or have already done so. But what is the situation in the food industries regarding the use of rapid methods? Are there efficient reliable rapid methods for the analysis of all hazards and if so, are they used in the food industries? For approaching the current status in the food industries, a survey was conducted in the framework of MoniQA project Towards the harmonization of analytical methods for monitoring food quality and safety in the food supply chain, in 17 countries (11 EU members and 6 non EU members) with more than 2600 questionnaires circulated to companies covering the whole food chain (raw material and ingredient suppliers, food processing companies, retailers, catering companies, etc.). From the questionnaires collected (661) the use of rapid, easy-to-apply methods, and online monitoring systems by the raw material manufacturers, food industries and retailers were assessed as well as the need and preparedness to implement such systems. According to the collected data 88% of the participants in the survey were certified to an externally accredited food safety management scheme, while 12% were not certified. Our results showed that raw materials and final products are the samples most routinely analyzed for microbiological contaminants, heavy metals, pesticides, foreign bodies, mycotoxins and allergens. Therefore, rapid methods for their analysis are either used or needed. Development of rapid methods is the major need of food companies, more specifically related to microbiological analysis, while food allergens and mycotoxins related test kits are also of high importance. Finally, regarding the use of automatic on/in line monitoring systems, companies use and need such systems mainly for temperature control, as temperature is a critical parameter for the safety of the product. In addition, automatic on/in line foreign body detection systems are also of high demand by various food companies. Concluding, the majority of the respondents considered that the introduction of rapid methods, including test kits, had contributed to improved food safety management. The results obtained indicate that companies in EU and non EU countries currently use or are well prepared for the implementation of new rapid methods of analysis and advanced monitoring systems. Keywords: HACCP, rapid methods, on/in line monitoring systems Acknowledgments The MoniQA Network of Excellence is funded by the European Commission (contract no. FOOD-CT2006-36337) within the Sixth Framework Programme Topic T5.4.5.1: Quality and safety control strategies for food (NOE). The authors appreciate the contribution of the following MoniQA partners: Hanna-Leena Alakomi, Anton J. Alldrick, Ayse Bakan, Aihnoa Bilbao, Zsuzsanna Bugyi, Cathrine Finne Kure, Velitchka Gotcheva, Sandra Kerbach, Susan Paulin, Xiaofang Pei, Maddalena Ragona, Christelle Robert, Kim Ahn To and Halina Tureskja.

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P31 MICROBIOLOGICAL AND HYGIENIC ASPECTS OF PINK COLORED NON SCABBY KERNELS L.Machikhina L.Lvova, L.Alexeeva Russian Scientific Research Institute for Grain of the Russian Academy of Agricultural Science (VNIIZ) 11, Dmitrovskoe Sh., Moscow, Russia

During some years wide-spread occurrence of pink-colored non scabby kernels is observed in a number of regions of Russia and the volume of damaged grains can reach hundreds thousand tons. Rye is a crop of elevated risk and the content of pink-colored grains in it is limited by the hygienic maximum level. Pink-colored kernels are more rarely found in wheat and barley grains also. But due to the absence of distinct visual differences and the objective methods of pink-colored non scabby kernels determination they may be confused with toxic scabby kernels, which leads to unjustified rejection of such lots. The aim of this research work was the ascertainment of microbiological and chemical nature of rye pink-colored non scabby kernels.Samples were taken during delivery of grain to elevator in NorthWestern, Central and Volga regions of Russia. Mycoflora of grains was analyzed using the plate method on agar media. In accordance with the preliminary data cousal fungi of pink-color may be Mycelia sterilia (wheat, rye) and Drechslera terres (barley).Pigments from grains and fungi culture were separated with acetonitrile and purified by means of column chromatography on silica gel. The final purification was carried out of highly officient preparative TLC. Nine components of pigments which were identified as methylanthraquionehydroxy derivatives were separated. Three prevailing components, by characteristics of mass spectra, absorption spectra in ultraviolet-visible region, 1HNMR spectra and melting points, were identified as chrysophanol, emodin and catenarin which are potentially toxic substances. These substances were also found in grain cultures (rye, barley, durum and soft wheat). The aims of further work are research of the toxicity level of pink-colored non scabby kernels and the development of analytical method determination of such grain. This trend secures both safety of grain and its rational use. Keywords: rye grain, pink-color of non scabby nature, Mycelia sterilia, pigments, emodin, chrysophanol, catenarin. Director of VNIIZ Dr.Lydia

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P32 RESISTANCE TO CIPROFLOXACIN AND TETRACYCLINE OF CAMPYLOBACTER SPP. ISOLATED FROM POULTRY PRODUCTS IN POLAND. E. Makiw D. Korsak, K. Rzewuska, K. Tomczuk, National Food and Nutrition Institute, Warsaw, Poland

Antimicrobial resistance has become a major public health concern in many countries. In the past decade, a rapidly increasing proportion of Campylobacter strains have developed resistance to the fluoroquinolones and tetracycline. The aim of the study was to determine susceptibility to fluoroquinolones (ciprofloxacin) and tetracyclines (tetracycline) among C. jejuni and C. coli strains isolated from poultry products. The food samples were collected from retail market in 2009. The isolation and identification of Campylobacter spp. were performed according to EN ISO 10272-1:2006. Species identification was confirmed by PCR. Campylobacter spp. susceptibility to ciprofloxacin and tetracycline was determined by broth microdilution method VetMIC Camp (SVA Uppsala, Sweden). Additionally all Campylobacter strains were screened for molecular mechanisms of the resistance to tetracycline and ciprofloxacin. The presence of tet(O) gene and Thr-86-Ile mutations in the gyrA gene were detected by PCR (1, 2). A total of 150 samples of poultry meat from retail market was taken for the analysis. Campylobacter spp. were found in 102 samples, what makes 68% of the number of samples. Campylobacter coli was isolated most frequently and its presence in 78 samples was demonstrated (76%), whereas Campylobacter jejuni was found in 24 samples (24%). The results of the research concerning antibiotic resistance revealed 100% C. jejuni and 97% C. coli ciprofloxacin resistant strains. Among tetracycline resistant strains 67% was estimated as C. coli and C. jejuni. Key words: termotolerant Campylobacter, susceptibility, fluoroquinolones, tetracyclines References: 1. Gibreel A., Tracz D.M., Nonaka L, Ngo T.M., Connell S.M., Taylor D.E., 2004, Incidence of antibiotic resistance in Campylobacter jejuni isolated in Alberta, Canada, from 1999 to 2002, with special reference to tet(O)-mediated tetracycline resistance. Antimicrob. Agents Chemother. 48:34423450. 2. Alonso R., Mateo E., Girbau C., Churruca E., Martinez I., Fernandez-Astorga A., 2004, PCRrestriction fragment length polymorphism assay for detection of gyr A mutations associated with fluoroquinolone resistance in Campylobacter coli. Antimicrob. Agents Chemother.. 48: 4886-4888.

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P33 OCCURRENCE OF CAMPYLOBACTER SPP. IN POULTRY PRODUCTS FOR SALE ON THE POLISH RETAIL MARKET E. Makiw D. Korsak, K. Rzewuska, K. Tomczuk National Food and Nutrition Institute, Warsaw, Poland

Thermotolerant Campylobacter species are the most common cause of bacterial gastroenteritis in humans. Consumption and handling of poultry meat are considered to be risk factors in acquired campylobacteriosis. The aim of the study was to identify main categories of the food of poultry origin known as the most significant reservoir of Campylobacter spp. The monitoring study was carried out in 20072008 in Poland. The food samples were collected from retail market. Altogether 912 samples were studied, including 443 samples of raw chicken meat, 146 samples of giblets and 323 ready-to-eat poultry products (spit-roasted chicken - 150, smoked chicken - 56 and pate, cold meats - 117). The studies were performed in accredited laboratories of 16 Provincial Sanitary and Epidemiological Stations and the National Food and Nutrition Institute pursuant to EN ISO 10272-1:2006. In the monitoring study Campylobacter spp. were detected in 51,7% of raw poultry meat. The differences were found between sub-groups consisted of the following chicken elements: wings, legs, corps (65,5 %), fillet (60,0 %) as well as raw ground poultry meat (15,1%).The prevalence of Campylobacter spp. in ground poultry meats was lower than in remaining sup-groups. In the raw poultry giblets Campylobacter spp. bacteria were found in 69 samples, that is in 47,3 %. A total of 323 ready-to-eat poultry products were studied. No contamination with Campylobacter spp. was found in any of the 206 studied samples of spit-roasted and smoked chicken. However, in two samples of pt (1.7 %) Campylobacter spp. were found. To summarize, our study, revealed that among all the poultry products, raw poultry meat and giblets were most frequently contaminated with Campylobacter. In spite of the high degree of contamination of the raw poultry meat, the ready-to-eat product has been recognized as safe. Key words: Poland, monitoring, termotolerant Campylobacter, poultry products References: Suzuki, H., & Yamamoto, S. (2009). Campylobacter contamination in retail poultry meats and byproducts in the world: a literature survey. Journal of Veterinary Medical Science, 71, 255-261. Whyte, P., McGill, K., Coweley, D., Madden, R.H., Morgan, L., Scates, P., Carroll, C., Leary, A. O., Fanning S., Collins, J. D., McNamara, E., Moore, J.E., & Cormican, M. (2004). Occurrence of Campylobacter in retail foods in Ireland. International Journal of Food Microbiology, 95, 111118.

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P34 THE EFFECT OF PREBIOTIC ADDITION AND INCUBATION pH TO SYNBIOTIC SOYGHURT M. Manullang J. R. Wijaya, N. Manthovani Food Technology Department, Faculty of Industrial Technology, Universitas Pelita Harapan, Indonesia (email : julia.wijaya@staff.uph.edu)

The development of soyghurt by the incorporation of probiotic as functional food has grown recently. Probiotic such as Bifidobacterium bifidum and Lactobacillus acidophilus has been proven to exert benefits to human host by modulating intestinal microflora. The aim of this research is to study the effect of prebiotic addition to the survival of probiotic Bifidobacterium bifidum and Lactobacillus acidophilus in different pH as indicated by Prebiotic Index Score. Synbiotic soyghurt was made by inoculating Bifidobacterium bifidum and Lactobacillus acidophilus into prebiotic supplemented soymilk o in anaerobic condition at 37 C for 12 hours. Different incubation pH (3.0, 3.8, 4.6, 5.4, 6.2, 7.0) were applied to simulate gastrointestinal pH. The prebiotic activity of inulin and hi-maize corn starch was measured by using Prebiotic Index. The growth and activity of Bifidobacterium bifidum and Lactobacillus acidophilus were determined by Total Plate Count, viscosity of the soyghurt, final pH, titratable acidity, total solid content and isoflavone content. The data showed that Bifidobacterium bifidum and Lactobacillus acidophilus were survived when incubated in pH 3.0 7.0. The supplementation of the prebiotic, inulin and hi-maize corn starch, into soymilk as synbiotic soyghurt improved the growth of Bifidobacterium bifidum and Lactobacillus acidophilus at low pH. The one with Hi-maize corn starch exhibited better prebiotic activity than inulin. The incubation of Bifidobacterium bifidum and Lactobacillus acidophilus in soyghurt supplemented with hi-maize corn starch at pH 6.2 exhibited the highest prebiotic activity as indicated by the highest Prebiotic Index score. Keywords: synbiotic, soyghurt, Prebiotic Index, probiotic, prebiotic

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P35 DIETARY EXPOSURE OF ACRYLAMIDE IN HIGH SCHOOL STUDENT GROUP L. Markov 1 1 1 K. Kukurov , Z. Ciesarov , P. imko 1 VP Food Research Institute, Bratislava, Slovak Republic 2 VUT University of Technology, Faculty of Chemistry, Brno, Czech Republic
1,2

ABSTRACT: The acrylamide is classified by IARC as a probably human carcinogenic agent. Due to a wide occurrence of acrylamide in many staple cereal and potato based foods the exposure to acrylamide cannot be omitted. This study is focused on the evaluation of the preliminary survey on the acrylamide exposure from foods with a supposed higher acrylamide level in the group of high school students in Czech Republic. This survey is based on the preliminary data from monitoring of acrylamide intake from foods in the Slovak and Czech Republics which revealed that the highest intake of acrylamide was observed in the group below 20 years (Markov et al., 2010). A frequency and a distribution of consumption of food during a day were ascertained by standardized interview using a questionnaire with partly free form questions. The data were evaluated depending on gender and time of day consumption. Moreover, the awareness of respondents about the occurrence of acrylamide in food was polled. KEY WORDS: acrylamide, exposure, thermally treated foods REFERENCES: IARC, 1994. Acrylamide. TA: IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans PG 1994; 60. Markov, L., Kukurov, K., Ciesarov, Z., imko, P., 2010. The acrylamide exposure from foods in the Slovak and Czech Republics. Potravinrstvo: 2010, 4, Special Issue, pp. 317-321. ACKNOWLEDGEMENT: This contribution is the result of the project implementation "Establishment of a HiTech centre for research on formation, elimination and assessment of contaminants in food" supported by the Research & Development Operational Programme funded by the ERDF. This work o was also supported by the Slovak Research and Development Agency under the contracts N . LPP 0310-09, SK-PL-0051-09, and VMSP-P-0089-09.

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P36 EVALUATION OF YEAST CONTAMINATION SOURCES IN TRADITIONAL IRANIAN YOGHURT DRINK (DOOGH) M. Mehraban Sangatash* 1,2 3 1,2 1,4 4 M. Sarabi Jamab , A.S. Vosough , R. Karagian , R. Nourbakhsh , F. Gholasi , 2 M. Mohsenzadeh 1 Food Science and Technology Research Institute, ACECR Mashhad Branch, Mashhad, Iran 2 Ferdowsi University, Mashhad, Iran 3 Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran 4 General Office Institute of Standard and Research Development, Mashhad, Iran *mehraban@acecr.ac.ir
Iranian yoghurt drink (Doogh) is a very popular and highly consumed product in Iran with a considerable increasing demand for its consumption. The annual production of plain Doogh in Iran was 14,400,000 tons in 2007. This study has been conducted to determine the yeast contamination sources during production of Iranian yoghurt drink in three local dairy plants, situated in Iran, mashhad. Samples were collected at Sixteen different control points, which involved every stage of production from the beginning to the end, including raw milk, pasteurized milk, fermented milk, doogh before and after pasteurization, doogh before and after packaging, samples from starter culture, salt, essential oil, potable water, washing water, production room air, swab test of PET, swab test of Nuzzeles, filling air and packaging air. Each plant was visited three times for sampling (occasions I, II and III).. Total count and yeast was determined according to the Institute of Standards and Industrial Research of Iran (ISIRI). We determined starter culture, potable water and washing water, Nuzzeles and packaging material and production room air as the possible contamination sources of yeast. It was shown that factories inevitably have various contamination sources because of the unstandardized design of yoghurt drink factories in Iran. It is necessary to determine critical control points in all factories and to organize autocontrol systems in order to eliminate or at least minimize the risk of contamination. Key words: Traditional Iranian yoghurt drink, yeast Contamination Sources, hygienic quality References: Avsar, Y.K., Karagul-Yuceer, Y., Tamucay, B., Kocak, C. and White, C.H. 2001. A comparative study on the production methods of Ayran, tradicional drinking yogurt of Turku. IFT Annaol Meeting. New Orleans, Louisiana. Corbo, M.R., Lanciotti, R., Albenzio, M. and Sinigaglia, M. 2001. Occurrence and characterization of yeasts isolated from milks and dairy products of Apulia region. International Journal of Food Microbiology. 69: 147-152. Gardner, I.A. 1997. Testing to Fulfill HACCP (Hazard Analysis Critical Control Points) Requirements: Principles and Examples. Journal of Dairy Science. 80:34533457. Gulmez, M., Guven, A., Sezer, C. and Duman, B. 2003. Evaluation of microbiological and chemical quality of Ayran samples marketed in Kars and Ankara cities in Turkey. Kafkas Univ. Vet. Fak. Derg. 9(1):49-52. Hedrick, T.I. and Heldman, D.R. 1969. Air quality in fluid and manufacturing milk products plants. Journal of Milk and Food Technology. 32: 265-269. Hekmat, M. and Lame, H. 1972. Dough, an Iranian milk product. Industries Alimentaires et Agricoles. 89: 17511752. ISO 6611.1992. Milk an milk Products- Enumeration of colony-forming units of yeasts and/or moulds-colonycount technique at 25C. Jakobsena, M and Narvhu, J 1996. Yeasts and their Possible Beneficial and Negative Effects on the Quality of Dairy Products, Review Article. International Dairy Journal. 6: 755-768. Kang, Y.J. and Frank, J.F. 1989. Evaluation of air samplers for recovery of biological aerosols in dairy processing plants. Journal of Food Protection. 52: 655-659. Loureiro, V. and quorol, A. 1999. the prevalence and control of spoilage yeasts in food and beverage. Trind in food science and technology. 10: 356-365 Moreira, S.R., Schwan, R.F., de Carvalho, E.P. and Wheals, A.E. 2001. Isolation and identification of yeasts and filamentous fungi from yoghurts in Brazil. Brazilian Journal of Microbiology. 32: 117-122. Suriyarachchi, V. R. and Fleet, G. H. 1981. Occurrence and growth of yeasts in yoghurts. Applied Environmental Microbiology. 42: 574-579.
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P37 CEREALS AND CEREAL PRODUCTS AUTHENTICITY ISSUES F. Melini M. Carcea National Research Institute on Food and Nutrition (INRAN), Via Ardeatina, 546, I-00178 Rome, Italy, carcea@inran.it

Food frauds are not a new phenomenon. They have been around a long time and already during the XIX century scientists pointed out the existence of food frauds related to food adulteration. Nowadays, food adulterations are more widespread and affect a broader range of food products. The number of known examples is, in fact, increasing more and more: substitution of ingredients, falsified packaging and labels, labelling to avoid import taxes and inspection, up-labelling to draw a higher price and product substitution. This poster aims at giving, first, a general overview of past and more recent frauds regarding adulteration of cereals and cereal products, i.e. bread and pasta. Socio-economic aspects and impacts of these frauds are also addressed, as, generally speaking, food frauds are not only dangerous for human health but are also bad for business. They destroy confidence in the food supply, alter consumer spending patterns, and destroy trust in specific brands and/or protected food products, such as the Italian PDO and PGI products pane di Altamura (Altamura bread), coppia ferrarese (Ferrara bread), farro della Garfagnana (Garfagnana emmer), farro di Monteleone di Spoleto (Monteleone emmer), etc. Subsequently, validated and non-validated methods that are currently used to determine authenticity 1 issues of cereals and cereal products will be presented (PCR, NIR, HPLC, PAGE, Real-Time PCR, H HR/MAS/NMR, capillary electrophoresis, etc.). Although further work will be required to confirm that non-validated methods are able to identify any degree of sample adulteration, they are, nevertheless, potential tools to consider and which control/food authorities may rely on to measure authenticity attributes and to tackle the issue. Gaps/needs to be addressed within the MoniQA Food Authenticity WG will be reported. A section of the poster will also be dedicated to legislation, both EU legislation which currently regulates the food authenticity of cereals and cereal products at the European level and also specific national regulations on this issue, such as the Italian law on cereals and products.

Keywords: food frauds, food authenticity, cereals, cereal products, socio-economic impact, validated methods, non-validated methods.

Poster Presentations

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P38 ANTIMICROBIAL ACTIVITY OF SUPERCRITICAL FLUID-CARBON DIOXIDE HOP EXTRACTS D. Mihaylova S. Bahchevanska, L. Kojuharova, V. Gotcheva, J. Stoikov Department of Biotechnology, University of Food Technologies, Plovdiv, Bulgaria

In the present study, the antimicrobial activity of hop extracts obtained by supercritical fluid extraction (CO2) was explored. Extraction was performed under various temperatures (18-40C) and pressure values (10.13-20.27MPa). The determination of antibacterial activity of the hop extracts was carried out by the agar-diffusion method against gram-positive and gram-negative bacteria. All extracts showed high antimicrobial activity - up to 31 mm zone. All tested concentrations of the hop extracts were inhibitory to Bacillus subtilis and Staphylococcus aureus. No inhibition of Pseudomonas aeruginosa was observed. Some of the extracts showed inhibitory activity towards Esherichia coli. The minimum inhibitory concentrations (MIC) of the extracts against Bacillus subtilis and Staphylococcus aureus were 1 mg% and less than 1mg%, respectively. Results show that the applied method is appropriate for extraction of hops biologically active substances with antimicrobial activity. Key words: hop, supercritical fluid (CO2) extraction, antibacterial activity References: He Guo- Ging, Xiong H., Chen Q., Ruan H., Wang Z., Traor L., 2005, Optimization of conditions for supercritical fluid extraction of flavonoids from hops ( Humulus lupulus L.), J. Zhejiang Univ Sci B.; 6(10): 9991004 Khan I. A., Mirza Z. M., Kumar A., Verma V., Qazi G. N. , 2006, Piperine, a Phytochemical Potentiator of Ciprofloxacin against Staphylococcus aureus, Antimicrobial Agents and Chemotherapy, Vol. 50, No. 2, 810-812 Langezaal C.R., Chandra A., Katsiotis S.T., Scheffer J.J.C., Haan A.B. de., 1990, Analysis of supercritical carbon dioxide extracts from cones and leaves of Humulus lupulus, J. Sci. Food Agric., 53, 455-463 Leal PF, Braga ME, Sato DN, Carvalho JE, Marques MO, Meireles MA., 2003, Functional properties of spice extracts obtained via supercritical fluid extraction, J Agric Food Chem., 23; 51(9): 2520-5. Oiye, S. O., Muroki, N. M., 2002, Use of Spices in Foods, The Journal of Food Technology in Africa, Vol. 7, Apr-Jun, pp. 39-44 Pessini G. L., Filho B. P.D., NakamuraC. V., D. A. G. Cortez, 2003, Antibacterial Activity of Extracts and Neolignans from Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) YunckAntibacterial activity , Mem. Inst. Oswaldo Crus, Rio de Janeiro, vol. 9 (8), 1115-1120

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P39 SIMULTANEOUS DETERMINATION OF DEOXYNIVALENOL, ZEARALENONE, T-2 & HT-2 USING DZT MS-PREP IN CONJUNCTION WITH LC-MS/MS C Milligan D Leeman E. Marley R-Biopharm Rhne Ltd. Block 10 Todd Campus, Acre Road, West of Scotland Science Park, Glasgow, Scotland, G20 0XA, United Kingdom.

Fusarium mycotoxins can be found in a variety of commodities when foods are stored under adverse conditions including temperature and humidity. Legislation for mycotoxins has recently increased to incorporate additional matrices and toxins, therefore resulting in an increased demand for faster and less labour intensive tests. Immunoaffinity columns are rapidly becoming the routine standard method of choice for the sample preparation of regulatory mycotoxin analysis however there is a growing need for multi-mycotoxin analysis using a single extraction method. In response, a multi-toxin immunoaffinity column (DZT MS PREP ) has been manufactured which enables the isolation and concentration of four commonly occurring Fusarium mycotoxins; deoxynivalenol, zearalenone, T-2 and HT-2. The advantages of this new immunoaffinity column are that only one sample preparation method is required for quantifying all four mycotoxins in a single run therefore having greater sample throughput and a reduction in the use of solvents and consumables. Both column repeatability and method repeatability were determined using various cereals including maize, wheat and oats. Extract repeatability data ranged from 92 - 109% (RSD 2.5 5.8%) with spiked maize, 74 103% revocery (RSD 2.1 8.0%) for wheat while recovery data for spiked oats ranged from 82 - 92% (RSD 2.3 4.3%). A range of FAPAS samples were also tested with all results falling within the specified ranges. DZT MS-PREP was shown to meet the European Commission analytical performance criteria for these mycotoxins, whilst providing a rapid and robust method of analysis enabling accurate, low level quantification of all four mycotoxins simultaneously.

Keywords Fusarium toxins ,immunoaffinity colums,mycotoxins

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P40 TRANS FATTY ACIDS LEVEL OF FRYING FATS IN POLAND H. Mojska K. Zukowska, K. Stos National Food and Nutrition Institute, Warsaw, Poland Trans isomers of unsaturated fatty acids are formed from cis unsaturated fatty acids by commercial partial hydrogenation during the processing of vegetable and fish oils and naturally during biohydrogenation by rumen microorganisms in the ruminants. Many studies showed that the consumption of trans isomers of fatty acids has been associated with increase of plasma LDLcholesterol and decrease HDL- cholesterol concentration. In the prevention of diet-related diseases it is recommended that trans fatty acids should provide no more than 1% of energy from the diet. The aim of our study was to determine trans fatty acids content in 64 samples of frying fats randomly taken at small outlets, fast food chain and restaurants all over Poland in 2008. The fatty acid methyl esters (FAME) were analyzed by high-resolution gas chromatography using HP 6890 gas chromatograph equipped with a split injector (split ratio 1:100) and MS detector. Helium was used as carrier gas (flow: 20cm/s) at head pressure of 43.4 psi. Separation was performed on a HP Sil 88 fused silica column (100 m x 0.25 mm i.d., film thickness: 0.20 mm). The oven temperature was programmed from 175 C for 40 minutes, thereafter 5 C per minute until 220 C, and held at this temperature for 16 minutes. Results are expressed as the weight percentage (% wt/wt) of total fatty acids detected with a chain length between 8 and 24 carbon atoms. Among the 64 tested samples of frying fats, 55% (35 samples) constituted oils and liquid frying fats. Other 45% constituted hydrogenated vegetables oils and special frying fats contain palm oil. The average content of trans fatty acids in all tested fats reached 1.10% and varied between below of limit of detection (LOD = 0.02%) to 33.21% w/w. We found significant (p< 0.05) higher trans fatty acids level in hydrogenated fats (1.97 % w/w) in comparision to liquid fats (0.39 % w/w). The highest trans fatty acids content was found in the sample of hydrogenated rape oil. Trans isomers of oleic acid were found in the highest amount in all frying fat samples making up 50% of total trans isomers in liquid fats and above 70% of total trans isomers in hydrogenated fats. Their levels varied from below 0.02 % to 6.25 % in liquid fats and from below 0.02 % to 33.21 % in hydrogenated fats. The average content of trans isomers of linoleic acids in liquid fats reached 0.09% of total fatty acids and was significantly (p<0.01) lower than in hydrogenated fats (0.46% wt/wt). Trans isomers of linoleic acid constituted above 20% of total t rans isomers in both groups. Comparing our results with TRANSFAIR Study results (1998) we found the lower average trans fatty acids content in frying fats, however in individual samples our results were higher. Key words: trans fatty acids, frying fats, GC/MS. References: 1. A. Aro, J. van Amelsvoort, W. Becker, M. A. van Erp Baart, A. Kafatos, T. Leth, G. van Poppel: Trans Fatty Acids in Dietary Fats and Oils from 14 European Countries: The TRANSFAIR Study. Journal of Food Composition and Analysis 1998, 11, s. 137 149. 2. H. Mojska, I. Gieleciska, J. Balas, M. Pawlicka, L. Szponar :Trans fatty acids in foods in Poland: Monitoring study. yw. Czow. Metab. 2006, 33, 2, s. 107 - 122. 3. H. Mojska, I. Gieleciska, L. Szponar, D. Marecka, M. Pawlicka: Izomery trans kwasw tuszczowych w produktach typu fast food. yw. Czow. Metab. 2007, 34, , s. 915 920.

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P41 DETERMINATION OF ACRYLAMIDE IN COFFEE BY LC-MS/MS AND ESTIMATION OF DIETARY EXPOSURE TO ACRYLAMIDE FROM COFFEE IN POLISH POPULATION H. Mojska I. Gieleciska National Food and Nutrition Institute, Warsaw, Poland

Acrylamide is a synthetic monomer, which has been used in production of polyacrylamides widely applied as a flocculants in drinking water and wastewater treatment, in paper processing and electrophoretic separation. In 1994, the International Agency for Research on Cancer (IARC) labelled acrylamide a probable human carcinogen. In food, acrylamide is formed as a result of a reaction between amino acids and reducing sugars during the heating of carbohydrate-rich foods. It occurs in high levels in potato crisps, French fries, cookies, cereals and in coffee. The purpose of our study was to determine the acrylamide level in twenty eight samples of roasted coffee, randomly selected from all over Poland in 2008-2009 and to assess the exposure of the Polish population on the acrylamide from coffee. The analysis of prepared coffee included a clean-up using solid-phase extraction (SPE) and detection by liquid chromatography tandem mass spectrometry (LC-MS/MS) using electrospray in the positive mode. d3-Acrylamide was used as an internal standard. For multiple reaction monitoring (MRM), the following MS/MS transitions were recorded: m/z = 72.1 > 55.2 and m/z = 72.1 > 44.1 for acrylamide with respective collision energies of 14 and 20 V, and m/z = 75.1 > 58.1 and m/z = 75.1 > 47.1 for d3-acrylamide with respective collision energies of 16 and 14 V. Dwell time was 150 msec. To quantify the acrylamide content in coffee we used area of ion peaks m/z 55.2 and m/z 58.1. The average level of acrylamide in roasted coffee was 179 g/kg and varied from 61 to 397 g/kg. Acrylamide content in the coffee infusion made using 5 g of ground coffee and 250 ml of boiled water was 3.6 g/L (range: 1.2 7.9 g/L). The comparison between acrylamide level in different types of roasted coffee was also done. To assess the Polish population exposure to acrylamide in coffee, food consumption data was taken from the Household Food Consumption and Anthropometric Survey in Poland (2000). The survey covered 4134 people from 1 to 96 years. The research method used was a 24-hours dietary recall carried out using Album of photographs of food products and dishes. The daily dietary acrylamide exposure was computed using a probabilistic approach for the total Polish population (1 96 years) and for the following groups: 1-6, 7-18 and 19-96 using a programme using Monte Carlo simulation technique. The estimated exposure of the whole Polish population on the acrylamide from coffee was 0.07 g/kg b.w./d and varied from 0.01 g/kg b.w./d in children and adolescents (7 18 years) to 0.09 g/kg b.w./d in adults (19 96 years). In our survey we found that roasted coffee was a significant source of dietary acrylamide intake, supply the total Polish population (1 96 years) 19% of acrylamide, with the highest amount in the adult population 27% of total dietary acrylamide intake. Key words: acrylamide, roasted coffee, LC-MS/MS, exposure.
References: 1. Castle L. Determination of acrylamide monomer in mushrooms grown on polyacrylamide gel. J. Agric. Food Chem. 41, 1261-1263, 1993. 2. Andrzejewski D., Roach J.A.G., Gay M.L., Musser S.M.: Analysis of coffee for the presence of acrylamide by LC-MS/MS. J. Agric. Food Chem., 52 (7), 2004. 3. Dybing, E. Sanner, T. Forum. Risk Assessment of Acrylamide in Foods. Toxicological Sciences 75, 7-15, 2003. 4. Ono, H., Chuda, Y., Ohnishi-Kameyama, M., Yada, H., Ishizaka, M., Kobayashi, H., Yoshida, M. Analysis of acrylamide by LC-MS/MS and GC-MS in processed Japanese foods. Food Addit. Contam. 20, 215-220, 2003.

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P42 IDENTIFICATION OF PEPTIDE MARKERS FOR CASEINATE BY LC- ESI - HIGH RESOLUTION MS: APPLICATION TO FINED WHITE WINES. L. Monaci I. Losito , F. Palmisano and A. Visconti. CNR, Institute of Sciences of Food Production (ISPA), Via Amendola 122/O Bari, Italy S.M.A.R.T. Inter-department Research Center, Department of Chemistry, University of Bari, Via E. Orabona 4, Bari, Italy.

Keywords: peptide markers, caseinate, fined wine, LC-ESI-MS, Orbitrap-MS Proteins are commonly present in wines at low levels, most of them having a remarkable technological and economical relevance. Interactions between proteins and wine components may also occur. Indeed, the formation of tannin-protein aggregates or protein-polyphenols complexes has been described. As, in general, proteins can affect wine stability and clarity, a variety of procedures have been developed for their removal from wines (Ferrerira et al. 2002). Fining agents typically used for clarification of wine are milk proteins, egg proteins and/or fish gelatine. Among milk proteins, caseins are universally known as suitable agents for binding phenolic compounds and reducing off-flavour ingredients that may affect wine taste and colour. Although it is assumed that fining agents are nearly quantitatively removed during the manufacturing process, to date there is no evidence that the consumer ready product is truly free of residues. In this regard, the analysis of fining agents, like caseins, eventually present as residues in wines is of paramount importance to safeguard the health of allergic individuals and to comply with the recent legislation issued in the field of food allergens. In particular, Directive 2007/68/EC has been issued by the European Commission to fulfil the need of labelling for food allergens. In the directive light is cast on all fining agents containing allergenic proteins and used for the manufacture of alcoholic and non alcoholic beverage; the declaration of their use, in the product label, is mandatory. To date ELISA has been the more widely method used for the detection of milk proteins in wines (Rolland et al, 2008). However, the LC-ESI-MS analysis of the tryptic digests of milk proteins extracted from wine could represent a complementary analytical strategy for their detection in those matrices. The present communication will then focus on the potentials and features offered by a method based on the coupling between LC and Orbitrap-MS in the identification of and casein tryptic peptides as markers of caseinate proteins in fined white wine samples. Thanks to the high resolution, mass accuracy and sensitivity provided by the Orbitrap technology, the method enables the detection and unequivocal identification, based on accurate mass values, of peptide markers for residual caseins arising from wine fined with caseinate in the 50-1000 g/mL range. The described approach appears to be a very promising tool for screening purposes as well as a confirmatory tool for the unequivocal identification of caseins in ELISA positive samples. References Ferreira, RB, MA Piarra-Pereira, S Monteiro, VB Loureiro, AR Teixeira. 2002. Tr Food Sci Technol 12: 230-239. Rolland, JM, E Apostolou, MP de Leon, CS Stockley, RE OHehir. 2008. J Agric Food Chem 56: 349354.

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P43 DYNAMICS AND BIODIVERSITY OF MICROORGANISMS (FUNGI AND YEAST) BY PCR-DGGE, WITH THE OBJECTIVE OF UNDERSTANDING OTA PRODUCTION ON COFFEE BEANS Montet Didier * 1,2 3 3 1 El Sheikha Aly , Suarez-Quiros Mirna-Leonor , Gonzales-Rios Oscar , Durand Nol 1. UMR Qualisud (CIRAD, Universit Montpellier II), 34095 Montpellier Cedex 5, France. 2. Department of Food Science and Technology (Minufiya University, Faculty of Agriculture), 32511 Shibin El Kom, Egypt. 3. Unidad de Investigacin y Desarrollo en Alimentos, Instituto Tecnolgico de Veracruz, 91860 Veracruz, Mexico. * E-mail address: didier.montet@cirad.fr
1

Ochratoxin A is a secondary metabolite produced by various filamentous fungi, is deemed to have nephrotoxic, immunotoxic, teratogenic and carcinogenic effects (1). In tropical zones, OTA is mainly produced in coffee beans by three Aspergillus species: A. carbonarius, A. niger section Nigri and A. ochraceus section Circumdati. In temperate zones Penicillium verrucosum and P. nordicum are known to synthesize OTA in food commodities (2, 3). The OTA content in coffee was shown to be closely link to harvesting conditions, post-harvest processing conditions and especially dry processing, storage and transportation conditions (4, 5, 6). In order to understand the OTA contamination process in foodstuffs, assays using PCR-DGGE (Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis) were carried. PCR-DGGE is a rapid molecular technique that was developed to monitor the dynamics of microbial populations (fungi, yeast and bacteria) by example in coffee mycoflora. In this work, the PCR-DGGE stages were optimized: extraction and amplification, repeatability and sensibility methodology applied to fungi were tested. Methodology Sensibility test was applied to selected Aspergillus species cultivated on Potato Dextrose Agar medium (AES, Combourg, France) for 5 days at 25C. The fungal spores were collected from plates in aseptic conditions using sterile distilled water with 0.1% Tween 80 solution. The suspensions of 7 2 fungal spores were quantified with Thomas cell and dilute in peptone water from 10 to 10 spores/mL DNA extraction method, according to El Sheikha et al. (7) different steps of extraction were used (mechanical/enzymatic/chemical). Additionally, the successful application of a eukaryotic universal primer for PCR permitted to amplify and identify many fungi species in one PCR step. The PCR products were analyzed by DGGE by using a Bio-Rad DcodeTM universal mutation detection system (Bio-Rad Laboratories, USA) using the procedure described by El Sheikha et al. (8). Conclusion The dynamics of fungus populations linked to OTA production, as well as post-harvest phytopathogens, could be studied by PCR-DGGE genetic fingerprinting. The advantages of this method are it efficiency on all microbial species (fungi, yeast and bacteria) and on the possibility of analysing a wide number of samples (30 samples) in a unique batch. References 1. IARC (International Agency Research of Cancer), 1993. Some naturally occuring substances: food items and constituents, heterocyclic aromatic amines and mycotoxins. IARC monographs on the Evaluation of Carcinogenic Risks to humans. IARC Press, Lyon, 56, 489-521.

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2. Pitt J.L., Basilico J.C., Abarca M.L., Lopez C., 2000. Mycotoxins and toxigenic fungi. Medical Mycology 38, 41-46. 3. OCallaghan, J., Caddick, M.X., Dobson, D.W., 2003. A polyketide synthase gene required for Ochratoxin A biosynthesis in Aspergillus ochraceus. Microbiology-SGM 149, 3485-3491. 4. Paulino de Moreas, M.H; Luchese R.H. Ochratoxin A on Green Coffee: Influence of Harvest and Drying Processing procedures. Journal of Agricultural and Food Chemistry, 2003. 51(19), 5824-5828. 5. Suarez-Quiroz M.; Gonzalez-Rios O.; Barel M.; Guyot B.; Schorr-Galindo S.; Guiraud., J-P. Effect of the post-harvest processing procedure on OTA occurrence in artificially contaminated coffee. International Journal of Food Microbiology. 2005. 103, 339 345. 6. Bucheli, P; Meyer, I; Pittet, A; Vuataz, G; Viani, R. Industrial storage of Robusta coffee under tropical conditions and its impact on raw material quality and ochratoxin A content. Journal of Agricultural and Food Chemistry, 1998. 46, 4507-4511. 7. El Sheikha, A., Condur, A., Mtayer, I., Le Nguyen, D. D., Loiseau, G. Montet, D. (2009). "Determination of fruit origin by using 26S rDNA fingerprinting of yeast communities by PCR-DGGE: An application to Physalis fruits from Egypt". Yeast, 26: 567573.. 8. El Sheikha, A., Mtayer, I., Montet, D. (2010). ""Determination of fruit origin by using 26S rDNA fingerprinting of yeast communities by PCR-DGGE: An application to Physalis fruits from Egypt". Food Biotechnology (Submitted).

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P44 EVALUATION OF THE EXTRACTION PROCEDURE FOR THE DETERMINATION OF DEOXYNIVALENOL IN MAIZE E. Numanoglu U. Uygun, V. Gokmen, H. Koksel Food Engineering Department, Hacettepe University, Beytepe, 06800, Ankara, Turkey

The effects of single and multiple stage extraction procedures on the extraction yield of deoxynivalenol (DON) from maize were studied. Naturally contaminated maize samples with different DON levels were used for analyses. In the multiple stage procedure, extraction of the ground samples was sequentially performed up to five times with water as the extraction solvent. The extraction yield of DON was determined for each stage. When the results obtained by single stage extraction were compared with the results from multiple stage extraction, there was a considerable difference between the extraction yields. The results revealed that a single stage procedure underestimated the level of DON in maize by a factor of up to 24% depending on the DON level. The extractability is an exponential function which can be used to optimize the multiple extraction conditions during the analysis of maize flour for DON. In general, two extraction steps were determined to be acceptable for the extraction of approximately 90% of DON. As a conclusion, two sequential extraction steps, each of 5 min, was decided to be suitable for the extraction of DON from maize. Keywords: deoxynivalenol, extraction, stagewise extraction, maize References: European Commission, 2007. Commission Regulation laying down the methods of sampling and analysis for the official control of the levels of mycotoxins in foodstuffs. 23.02.2006, EC No. 401/2006. European Commission, 2007. Commission Regulation amending Regulation (EC) No 1881/2006 setting maximum levels for certain contaminants in foodstuffs as regards Fusarium toxins in maize and maize products. 28.09.2007, EC No. 1126/2007. Gokmen, V., Morales, F.J., Atac, B., Serpen A., Arribas-Lorenzo, G., 2009, Multiple-stage extraction strategy for the determination of acrylamide in foods, Journal of Food Composition and Analysis, 22, 142147. Krska, R., Baumgartner, S., Josephs, R., 2001, The state-of-the-art in the analysis of type-A and -B trichothecene mycotoxins in cereals, Fresenius J Anal Chem, 371, 285 299. MacDonald, S.J., Chan, D., Brereton, P., Damant, A., Wood, R., 2005, Determination of Deoxynivalenol in Cereals and Cereal Products by Immunoaffinity Column Cleanup with Liquid Chromatography: Interlaboratory Study, J. AOAC Int., 88, 1197. Spanjer, M.C., Rensen, P.M., Scholten, J.M., 2008, LCMS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs, Food Additives and Contaminants, 25(4), 472489. Sulyok, M., Berthiller, F., Krska, R., Schuhmacher, R., 2006, Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize, Rapid Communicatons in Mass Spectrometry, 20, 26492659.

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P45 INVESTIGATION OF THE VARIOUS DERIVATIZATION METHODS INCLUDING TRIMETHYLSILYLATION OF MELAMINE, AMMELINE, AMMELIDE AND CYANURIC ACID FOR THE SIMULTANEOUS GC-MS ANALYSIS C.H. Oh* a b c J.E. Yoon, S.H. Kim, S.H. Cheon, H.J. Lee, J.H. Hahm, J.S. Roh , K.S. Kim , J.W. Park , c J.H. Woon Dept. of Oriental Medical Food and Nutrition, Semyung University, 117 Semyung-ro, Jecheon-city, Chungbuk 390-711, South KOREA a Safety Guarantee Institute, HAITAI Confectionery & Foods Co., Ltd., Seoul, South KOREA b Dept. of Food & Nutrition, Chosun University, 375 Seosuk-dong, Gwangju 501-759, South KOREA c National Veterinary Research & Quarnatine Service, 480 Anyang 6-Dong, Manan-Gu, Anyang-Si, Gyungido 430-824, South KOREA *Corresponding Author (och35@semyung.ac.kr)

The scandal, in which melamine was added to raw milk to make it appear higher in protein, led to the deaths of six babies and made some 300,000 ill. Melamine co-exposure with the other analogues such as cyanuric acid can induce acute melamine-analogue crystal nephropathy, leading to renal failure at much lower doses than with either compound given individually. Even though there are various methods currently in use for the screening, confirmation and quantification of melamine and its analogues, they have not been evaluated sufficiently. Liquid chromatograph with tandem mass spectrometer (LC-MSMS) is recommended for the analysis of melamine. However LC-MSMS is still not common to ordinary food analysis laboratory due to the high expense. Therefore a various derivatization methods of melamine, ammeline, ammelide and cyanuric acid (melamines) were investigated before GC-MS analysis that must be quite common instrument could be maintained with low expense. Ethylchloroformation, trifluoroacetylation and trimethylsilylation (TMS) were tested. TMS derivatization was optimized for the TMS reagents, solvents, the ratio of TMS reagent and solvent, heating time, stability of the derivatives etc. for the GC-time of flight mass spectrometry. Among the three derivatization methods tried, only TMS worked for the simultaneous derivatization of the four melamines. TMS derivatives of the four melamine related compounds were reliable for the quantitative analysis if two hours reaction at 90 C (inside of vial reached to 70 C) was applied after ultra-sonication for the through mixing of the whole reagents inside the vial insert. The reasonable LOQ level was 0.05ug/mL (with 2,6-diamino-4chloropyrimidien as I.S.). The derivative was stable up to two days (or further) after the derivatization. The optimized derivatization method was applied to the analysis of some stock farm product for the melamines. Key Words : melamine, cyanuric acid, ammeline, ammelide, derivatization, GC-MS

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P46 POPS (PERSISTENT ORGANIC POLLUTANTS) IN SOME FISH SPECIES FROM THYRRENIAN AND ADRIATIC SEA E. Orban M. Masci National Institute for Food and Nutrition Research, via Ardeatina 546 00178, Rome (Italy) Samples of European hake (Merluccius merluccius), European Cuttlefish (Sepia officinalis), European pilchard (Sardina pilchardus), Red mullet (Mullus barbatus), fish species present throughout the Mediterranean and most common for the Italian fishery, both from Thyrrenian and Adriatic Sea, were analysed for POPs content. The presence of organochlorine pesticides and polychlorinated biphenyls (PCBs) in the aquatic environments represents one of the most debated environmental questions due to their ubiquitous presence, accumulation in the food chain and incidence on the public health .Chemicals named POPs are largely employed in the past. Most of them are now banned for use. The Stockholm Convention (2001), now in force, recognized an initial list of twelve Chlorine-based POPs, or classes of POPs, as causing adverse effects on humans and the ecosystem. They were placed in three categories: Pesticides (such as DDT), industrial chemicals (such as Polychlorobiphenyls) and by-products (such as Dioxins). In the year 2009 the Convention listed nine additional chemicals (Chlorine-, Fluorine- and Bromine-based) as persistent organic pollutants.In our Laboratory 52 different POPs are currently monitored in fish products. With a decades experience in the determination of POPs, much attention is paid by us that these difficult analyses at trace levels are executed in compliance with GLPs (Good Laboratory Practices). Quality controls and Method Validation are achieved by participating in interlaboratory proficiency tests. The use of certified pure standards and the analysis of certified reference materials are also important steps in the method quality assuring. The 2002/657 European Community Decision concerning the performance of analytical methods in monitoring certain substances in animals and animal products is always taken into account. According to most methods currently used two main steps have to be performed in our work: sample preparation and instrumental analysis. Instrumental analyses are achieved by using a double Gas Chromatography determination, the first with ECD detector and the second with MS and MS/MS technique. Results shows low contamination levels in the muscle tissue, well below the currently available tolerance values (the maximum observed concentration for total DDT was 11 ng/g edible portion). Nevertheless we must emphasize that tolerances can be regarded as agreement values being fully safe only the level of zero ng/g and being partially unknown the impact on health of these chemicals, even at trace levels. Moreover it must be noted that does not exist a limit for the sum of total Organochlorine pollutants in a sample (this is a lack in most current regulatory actions). Finally the human bioaccumulation should be considered: consumption of low contaminated foodstuff during many years could lead to a great uptake of these dangerous Xenobiotics. In any case the presence of POPs was evident in all samples we analyzed, so confirming their global presence in the food chain. KEYWORDS: Pesticides, Polychlorobiphenyls, Fish from fishery

Orban E. and Masci M., Pollutant levels in Fish and Fishfeed from Aquaculture in Aquaculture Europe 2002 Conference, Trieste October 16-19 2002, Book of Abstracts 2002, 400-401 Orban Elena and Masci Maurizio, Residue Level of Organochlorine Pesticides in Fish from Italian Fishery and Aquaculture in Progress in Pesticides Research, chapter 5 335 -369 Nova Science Publishers, New York (2008) Stockholm Convention on Persistent Organic Pollutants (http://chm.pops.int/) FAPAS Proficiency Test Report, series 5, Round 32, June-July 2003, Sand Hutton, York (UK)

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P47 VALIDATION PROTOCOLS FOR FOOD ANALYSIS METHODS. MEASUREMENT UNCERTAINTY AND VALIDATION CRITERIA V. Oreopoulou T. Tsironi, M. Tsevdou Laboratory of Food Chemistry and Technology, School of Chemical Engineering, National Technical University of Athens (ftsironi@chemeng.ntua.gr)

It is internationally recognized that validation of methods is necessary in analytical laboratories. The use of validated methods is important for an analytical laboratory to show its qualification and competency. Validation is the tool used to demonstrate that a specific analytical method actually measures what it is indented to measure and thus that it is suitable for its intended purpose. The objective of the present study was the collection, survey and review of method validation protocols generated by standardization organizations, other nationally and regionally significant bodies, organizations and companies. Criteria for acceptance of method performance characteristics were also collected and surveyed and gaps and needs were addressed. The aim was to bring together the different approaches and to make definite recommendations as to the best practice. Food analysis laboratories use mainly the validation protocols published by ISO, IUPAC and Eurachem. ISO and Eurachem provide detailed guidelines about measurement and sampling uncertainty. There are not significant deviations between protocols, while the main performance criteria are accuracy - as expressed by precision under repeatability and reproducibility conditions, trueness and bias, recovery, and robustness - sensitivity, limit of detection, and limit of quantification. The need of estimation of the uncertainty of a measurement is emphasised, including, among other sources, the uncertainty in sampling. Gaps were noticed on validation of qualitative methods and with methods for the determination of food allergens, mainly due to lack of certified reference materials. Additionally, some issues are needed to be harmonised for intra-laboratory validations (agreement on a common protocol for single laboratory validation, use of the same parameters for method validation, use of the same criteria for acceptance or rejection of validation parameters). Specific guidelines have been published for the validation of microbiological methods. The major gap and research need on the application of these guidelines is that they are not referred to the estimation of measurement uncertainty. Gaps also arise in some applications (e.g. PCR methods for the identification of Campylobacter spp.). Keywords: Method validation, food analysis, validation protocols

Acknowledgments The MoniQA Network of Excellence is funded by the European Commission (contract no. FOOD-CT2006-36337) within the Sixth Framework Programme Topic T5.4.5.1: Quality and safety control strategies for food (NOE). The authors appreciate the contribution of the following MoniQA partners: Paula Alvito, Sandra Kerbach, Susan Paulin, John-Erik Haugen, Christoph Von Holst, Andreas Hhl, Marina Carcea, Bert Poepping, Hans van Egmond, Martin Rose, Roy Macarthur.

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P48 APPLICATION OF CELLULAR TOXICITY ANALYSIS METHOD IN VITRO ON RED FRUIT (Pandanus conoideus Lam) EXTRACT Palupi NS 1) 2) Zakaria FR dan Sukirno 1) Deparment of Food Science and Technology, Faculty of Agricultural Engineering and Technology, 2) Bogor Agricultural University; National Food and Drug Control Agency, Indonesia
1)

Plants generally contain antinutritive or toxic compounds that can be dangerous if consumed by humans. These compounds can be eliminated such as through the treatment process. Red Fruit (Pandanus conoideus Lm) is one of the plants that are consumed by people in the eastern region of Indonesia, especially Papua Island. The red fruit oil extracted using traditional technology to get oil by its boiling at a high temperatures and for a long time periods, then the red fruit oil extract is used as a sauce and eaten with a sweet potato. This research was conducted to test the safety of red fruit oils and extracts, using cellular toxicity method in vitro. Red fruit used in this research is a red fruit oil obtained by traditional extraction methods and red fruit extract the results of water, hexane and methanol extraction. Cellular toxicity test conducted using lymphocytes isolated from human blood. Lymphocyte cells were cultured using the red fruit oil, aqueous, hexane and methanol extract with three concentrations, ie 8.3, 33.3 and 66.7 ug / ml. o Inkuubasi performed using a CO2 incubator at 37 C, for 72 hours. Lymphocyte proliferation and cell death were determined using MTT (3-(4,5-di-methylthiasol-2-yl)-2-5-diphenyl-tetrazolium) and stained using blue tryphane, and the EC50 (effective concentration-50) was calculated. The results showed that the cell death of cultured lymphocytes was decreased by aqueous extract of 6 6 red fruit from 1.3x10 cells/ml to 0.2x10 cells/ml. Whereas the number of lymphocyte cells death 6 cultured with red fruits oil, methanol and hexane extract decreased respectively: from 1x10 to 6 6 6 6 6 0.4x10 cells/ml; from 0.8 to10 -0.6x10 cells/ml, and from 0.9 to 10 -0.7x10 cells/ml. While the proliferation of cells lymphocyte cultured with water extract, red fruit oil, methanol and hexane extract at a concentration of 8.3, 33.3 and 66.7 ug/ml were respectively: 50, 120 and 220%, 40, 50 dan140%; 60, 110 and 105%, and 40, 45 and 60%. At all concentrations tested, the red fruit oil and extract showed no toxic effect. Keyword: cellular toxicity, lymphocite, in vitro, pandanus conoideus Lam, EC50

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P49 RISK OF ACCUMULATION OF ATTENDANT PHARMACEUTICAL RESIDUES IN WASTE WATERS FOR FISH MEAT Zs. Gy. Papp, Zs. J. Sndor and I. Csengeri Research Institute for Fisheries, Aquaculture and Irrigation, Szarvas, Hungary

Intensive use of different xenobiotics including pharmaceuticals is increasing concentration of persistent pollutants in the natural and waste waters as well as sediments. Among others, the contaminant effect of medicines and endocrine disruptors has become evident only in the last years (Colborn at. al. 1996). By the reason of their very low and hardly detectable concentration, monitoring of these contaminants were previously neglected and often considered harmless, however most of these compounds have significant effect for the environment. Comprehensive research of these xenobiotics has become possible due to the more up-to-date analytical methods and instruments only in the last years. There are several compounds, like antibiotics, which cannot eliminate entirely by wastewater purification and thus, due to their continuous supply, these compounds become persistent in the environment (Kmmerer, 2008). Interaction and accumulation of these xenobiotics in the different aquatic organisms are mostly unknown (Epel, 1998). Meat of fish might mean risks even if that originated from aquaculture or natural water. According to these observations our aim was to study the accumulation of pharmaceuticals especially some antibiotics (oxitetracycline, flumequine, sulphonamide and nitrofurans) in waste water, sediment and fish tissues. Combination of screen (ELISA) and HPLC technique was applied for determination of different antibiotic compounds, metabolites and residues. Our results have shown that concentrations of different antibiotics and metabolites in different Hungarian waters and sediments represent average values, so currently the risk of the accumulation is likely low. However application of some antibiotics (like oxitetracycline (OTC)) used in aquaculture might have high risk. In our model -1 experiment OTC concentration was found about 10 times over than MRL (100 g kg ) in the meat of common carp some weeks after stopping the administration of the drug. FLU concentrations declined rapidly to the MRL (600 g kg-1). This data suggest that FLU might use securely common carp farming. This research was partially supported by European Union grant (FP6 FOOD-CT-2006-16249 Project AQUAMAX) KEYWORDS: xenobiotics, pharmaceutical residues, antibiotics, waste water, fish REFERENCES Colborn, T. Dumanoski, D. and Peterson, J., 1996. Our stolen future: Are we Threatening our fertility, Inteligence and survival? A scientific detective story. NEW York, NY: Dutton, Penguin Books VSA, 1996. Epel, D. 1998. Use of multidrugtransporters as first lines of defuse against toxin sin aquatic organism. Comp. Biochem and Phys. A. 120(1), Kmmerer, K. 2008. Pharmaceuticals in the Environment. Sources, Effects and Risk, third ed. Springer, Berlin, Heidelberg.

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P50 ICC YOUR CONNECTION TO THE WORLD OF CEREALS Roland Ernest Poms ICC International Association for Cereal Science and Technology, Vienna, Austria, www.icc.or.at

ICC - The International Association for Cereal Science and Technology is the pre-eminent international association in the field of cereal science and technology committed to the international cooperation through disseminating information and developing standard methods for the well-being of all people. ICC was founded in 1955, ICC is an independent, internationally recognized organisation; ICC is a non-political, non-profit-making, neutral forum for food scientists and technologists. ICC is an important contributor and publisher of International Standard Methods for food quality and safety, many of which were adopted by ISO, CEN, and other international organisations. Standardisation work and method validation is coordinated by the ICC Headquarters; the actual research and communication activities of ICC are for the most part carried on by more than 40 Working and Study Groups and several Task Forces (e.g. mycotoxin analysis, sampling, food allergens). ICC is an important organizer of national and international events (e.g. congresses, workshops). ICC is a promoter of international cooperation on a global, regional and national level and a mediator between science and technology in research and practice. At present more than 50 countries are represented in ICC. ICC has consultative status or is affiliated to the following organisations: WHO, FAO, UNIDO, ISO, CEN, AOAC, AACC Intl, AOCS, CCOA, a. o. Examples of ICC publications are Web based information material at www.icc.or.at, ICC Standard Methods, International Journal of Quality Assurance and Safety of Crops & Foods, monthly eNewsletter, Multilingual Dictionary, Who is Who in the World of Cereal Science, Conference Proceedings, Conference Books, etc. ICC is successful co-ordinator (e.g. ECC 2002; MoniQA NoE www.moniqa.org), and is partner in several national and international research projects (e.g., HEALTHGRAIN, www.healthgrain.org; ISEKI Food-2 http://www.esb.ucp.pt/iseki/; PlantLibra FP7). ICCs world-wide activities, covering all continents, are coordinated by its Vienna-based Headquarters (www.icc.or.at).

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P51 QAS QUALITY ASSURANCE AND SAFETY OF CROPS & FOODS Roland Ernest Poms 2 Stanley Cauvain 1. ICC International Association for Cereal Science and Technology, www.icc.or.at, Austria, CoEditor in Chief of QAS 2. Baketran, www.baketran.com, United Kingdom, Co-Editor in Chief of QAS
1

ICC International Association for Cereal Science and Technology (www.icc.or.at) - in cooperation with MoniQA (www.moniqa.org) has launched a new international journal titled QAS - Quality Assurance and Safety of Crops & Foods, www.icc.or.at/journal/qas. QAS is the Official Journal of ICC. It is a peer-reviewed scientific journal published by Wiley-Blackwell with Stanley Cauvain (BakeTran, UK) and Roland Poms (ICC, Austria) as the Joint Editors in Chief. The first issue appeared in March 2009. The journal publishes research papers from around the globe with a major focus on the research and development of new analytical tools for assuring the quality and safety of cereals, crops and derived foods and feeds, as well as refined products and ingredients, to ensure traceability and authenticity and to contribute to food security. The journal further describes the technologies associated with harvesting, storing, processing and manufacturing of crops and foods, feeds, ingredients and other refined products. The scope of publications covers also other disciplines related to food and feed quality and safety, such as legal requirements, regulations, socio-economic impact, globalisation and trade, food security, international organisations and agreements. The aims of the journal are to publish research papers from around the globe to publish peer-reviewed results of validation studies of analytical methods to publish reviews in the food safety and quality assessment areas to promote new technologies in food production, storage, processing and analysis to inform about new regulations, political decisions, and agreements affecting global trade of cereals and other crops, foods and feeds to inform about relevant meetings and training opportunities, educational programmes and association news to report on various meetings and workshops to report on relevant research projects (e.g. MoniQA NoE coordinated by ICC, HealthGrain, ISEKI Food-2, and others, ) to inform about publishing opportunities to promote new analytical methods, new testing devices, analytical assays and appliances to present new publications (books, protocols, standards,) to give a voice to all Country Members and Corporate Members of ICC, partners of the MoniQA NoE and members of a tentative future entity MoniQA2, and the journal subscribers; to publish the work and activities of the ICC technical working groups and MoniQA working groups;

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P52 RAPID METHODS FOR FOOD ADDITIVES: PRESENT STATUS, GAPS AND NEEDS! V. Psimouli C. Zerva, V. Oreopoulou Laboratory of Food Chemistry and Technology, School of Chemical Engineering, National Technical University of Athens, Greece

The effect of additives is not only limited to the quality but also to the safety of the final product, since in most cases the content of the additive should not exceed the respective maximum level. Hence the effectiveness of analytical methods for their determination is of utmost importance. Furthermore there is increasing need for rapid and high-throughput methods for multicomponent analysis which can be used e.g. for screening purposes and are therefore beneficial for food industries as well as for authorities. Rapid methods for the analysis of food additives appear to be limited. Multicomponent methods are available for the analysis of some sweeteners whereas test kits are available for the analysis of nitrates. In particular HPLC, ion chromatography and capillary electrophoresis are suitable tools for the multi-component analysis of artificial sweeteners. Furthermore rapid methods for the determination of one or two sweeteners (cyclamate and saccharin, acusulfame-K and aspartame) have been reported. Flow-injection analysis, electrochemical and spectroscopic methods are less time consuming and present comparable efficiency compared to the multi-component ones in terms of sensitivity and accuracy. The present work provides an overview of available rapid methods for food additives and the identification of gaps and needs for the development of new ones. The collection of these data was accomplished by the contribution of partners with expertise in the field of methods of analysis on certain food additives. Acknowledgement th The funds for this research were provided by the EU 6 Framework Network of Excellence Project MoniQA Keywords: food additives, rapid methods, multicomponent analysis References 1. V. Oreopoulou et al. Assessing food additives: the good, the bad and the ugly. Quality Assurance and Safety of Crops and Foods, 1,2,2009 pages 101-110 2. A. Zygler, A. Wasik, J. Namiesnik. Analytical methodologies for determination of artificial sweeteners in foodstuffs. Trends in Analytical Chemistry, Vol. 28, No.9, 2009

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P53 SURVEY: OCHRATOXIN A IN WINES (VQPRD) FROM THE NORTH WESTERN GREECE (ACHAIA, ILEIA) Y. Sarigiannis 2 1 2 1 1 S. Agriopoulou , A. Koliadima , J. Kapolos , T. Tsegenidis , G. Karaiskakis 1 2 Toxin Analysis Lab ELETOX, University of Patras, TEI Kalamatas, Department of Food Technology
1

Ochratoxin A (OTA) is a mycotoxin produced mainly by several fungal species of the genera Aspergillus and Penicillium. This mycotoxin has been shown to be nephrotoxic, hepatotoxic, teratogenic and carcinogenic to animals and has been classified as a possible carcinogen to humans. OTA contaminates wine all over the world, and its consumption may signi cantly increase human exposure to this toxin. OTA presence in wines is due to the black aspergilli, mainly A. carbonarius, which can grow on grapes in the vineyards and produce the toxin. At grape crushing, the juice can be contaminated with the toxin which is carried over into wine, where it persists due to its stability. Generally, red wines seem to contain a higher amount of OTA than white or ros wines, and some results suggest that, at least for European and North African cultivation areas, southern regions are more affected by the contamination problem. Differences are attributed to climatic factors, winemaking techniques grape cultivation and storage conditions. The climatic and geographic differences affect mould development and OTA contamination of grapes, and consequently, the wine. Generally, in the southern wine-growing region sampled, highest OTA occurrences and levels were observed. Overall, wine is important to the European economy and populations and, therefore, is important to assure that it is free of harmful contaminants. Currently, several countries have specic regulations for OTA in various commodities, 2 g/kg OTA being the maximum level allowed for wine, grape must and grapes in the European Union (Commission Regulation (EC) No.1881/2006) . The aim of this study was to assess OTA occurrence in VQPRD wines from the North Western Greece (Achaia, Ileia). A total of 20 wine samples originating from different regions of North Western Greece were analysed for OTA. An analytical method based on immunoaffinity column (IAC) for clean-up and high performance liquid chromatography with fluorescence detection (HPLC-FD) was used to determine OTA in wines. 65% of the wines sampled contained OTA (13 out of 20) with 92% much lower than 2 g/L OTA. Although, OTA is present at high levels in red wine according to previous authors, 60% (3 out of 5) of the tested red wines werent contaminated and the two others were contaminated at limit of quantification. On the other hand, 64% of the dry white wines (7 out of 11) were contaminated at low levels of OTA, much lower than the European MRL. All the sweet wines (four samples of white muscats and one red wine of mavrodaphne) were contaminated. Only one sample of sweet wine (Muscat Rio Patras) was contaminated above the 2 g/L OTA. KEYWORDS: Food Safety, Mycotoxins, Ochratoxin A, Aspergillus Carbonarius, Red Wine References
Commission Regulation (2006). (EC) No 1881/2006. Brussels: Ocial Journal of the European Union. Bell, N., Marn, S., Duaigues, A., Ramos, A. J ., & Sanchis, V. (2004). Journal of the Science of Food and Agriculture, 84, 591594. Chiodini, A. M., Scherpenisse, P., & Bergwer, A. A. (2006). Journal of Agricultural and Food Chemistry, 54, 73997404. Blesa, J., Soriano, J. M., Molto, J. C., & Manes , J. (2004), Journal of Chromatography A, 1054, 397401. Pietri, A., Bertuzzi, T., Pallaroni, L., & Piva, G. (2001). Food Additives and Contaminants, 18, 647654. Ottender, H., & Majerus, P. (2000). Food Additives and Contaminants, 17, 793798. Visconti, A.; Pascale, M.; Centonze, G. (1999) J. Chromatography A, 864, 89-101. Visconti, A., Pascale, M., Centonze, G., (2001) J. AOAC Int, 84, 1818-1827.

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P54 SURVEY OF PHTHALATE LEVELS IN FOOD ON THE BELGIAN MARKET K. Servaes N. Hufkens, M. Van Holderbeke, L. Geerts, G. Vanermen Flemish Institute for Technological Research, Boeretang 200, 2400 Mol, Belgium

Dialkyl and mono-alkyl phthalate esters, hereafter referred to as phthalates, form a group of chemical compounds that are widely used as plasticizers due to their flexible and weak character. In Western Europe approximately one hundred different phthalates are available on the market, of which di-2-ethylhexyl phthalate (DEHP), diisononyl phthalate (DiNP) and di-isodecyl phthalate (DiDP) are the most dominant. Due to their widespread use, phthalates are ubiquitous in the environment. Human exposure to phthalates occurs predominantly via dietary intake. Phthalates are thought to be responsible for adverse health effects in humans, like disruption of the endocrine system. The presence of phthalates in food is not only because of their movement up the food chain, but also due to their migration from packaging and contact materials mostly into lipid-rich foodstuffs. In this study, the presence and concentration of phthalates in food products on the Belgian market were determined and the various contamination pathways were explored. For this purpose, a sampling and measurement campaign of phthalate levels in food on the Belgian market has been set up. This work is part of a larger study aiming at the quantification of the dietary intake of phthalates for the Belgian population. Four hundred representative samples of widely consumed foods were purchased in Belgian shops. Sample selection is based on 1) consumption data from the Belgian national food consumption survey (2004) and 2) the likelihood that foodstuffs contain phthalates. Brand name, packaging material and properties, fat content, date of production or shelf life, time and place of purchase, picture and - if relevant - product specific properties (e.g. pH, preserving agent) were stored in a database. Due to the omnipresence of phthalates in the laboratory environment, precautionary measures have been taken to control blank concentrations. The measurement campaign involves the determination of ten phthalates in the selected foodstuffs: dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP), di-isobutyl phthalate (DiBP), benzylbutyl phthalate (BBP), di-2-ethylhexyl phthalate (DEHP), dicyclohexyl phthalate (DCHP), di-n-octyl phthalate (DNOP), di-isononyl phthalate (DiNP) and di-isodecyl phthalate (DiDP). Suitable extraction techniques have been developed for the various food matrices. Instrumental analysis is performed by means of gas chromatograph-mass spectrometry with electron impact ionization (GC-EI-MS). The phthalates DiNP and DiDP consist of a cluster of isomers, which hampers an accurate determination of these compounds by GC-MS. As DiNP and DiDP are replacing the more toxic DEHP, their use is still growing. Missing data of DiNP and DiDP will consequently influence the general overview of contamination of foodstuffs with phthalates. By applying LC-MS/MS, however, in combination with the appropriate analytical column, the different isomers of DiNP and DiDP coelute into a single peak. The results of the campaign show a wide variety of phthalate levels in food of the Belgian market, depending e.g. on pH and fat content of the food product. Contamination of food can occur from various sources such as migration from packaging material (including printing inks and glues), contact with plastics during food production and preparation, and transfer via the environment. We explored the possible impact of the different contamination sources by combining the information on product properties with the results of the analytical measurement campaign.

Keywords: phthalates, processing contaminants, food products, contamination pathways, analytical measurement Wormuth, M.; Scheringer, M.; Vollenweider, M.; Hungerbhler, K. Risk Analysis 2006, 26, 803-824. Dickson-Spillmann, M.; Siegrist, M.; Keller, C.; Wormuth, M. Risk Analysis 2009, 29, 1170-1181. Lin, Z.-P., Ikonomou, M.G.; Jing, H.; Mackintosch, C.; Gobas, F. Environ. Sci. Technol. 2003, 37, 2100-2108. Srensen, L.K. Rapid Commun. Mass Spectrom. 2006, 20, 1135-1143.

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P55 OCHRATOXIN-A PRODUCING MOULDS AND OCHRATOXIN A IN FORAGES M. krinjar 2 3 1 D. Jaki-Dimi , D. R. Milievi and N. Nemet 1University of Novi Sad, Faculty of Technology, Novi Sad, 2Institute of Veterinary Medicine of Serbia, Belgrade and 3Institute of Meat Hygiene and Technology, Belgrade, Serbia
1

Mycological and mycotoxicological analyses of some forages (silage 21 samples, dried clover 27 samples, fresh clover 3 samples, hay 10 samples, dried cornstalks 6 samples, pelleted sugar beet pulp 12 samples, fresh sugar beet pulp 1 sample, malt spent grains 6 samples, fresh rape leaf 1 sample) throughout one research year (in all seasons) were done. A special attention was paid to the presence and frequency of ochratoxin A-producing moulds and ochratoxin A (OTA). Mycological examinations were carried out according to the modern mycological methods. Determination of OTA was done using a fluorometric method (OchraTestTM Instruction Manual, 1997, VICAM, USA). About 93% of forage samples were contaminated with moulds at various degree. Isolated fungi were classified even into 28 genera (Absidia, Acremonium, Alternaria, Aspergillus, Aureobasidium, Botrytis, Chaetomium, Cladorrhinium, Cladosporium, Fusarium, Geotrichum, Gilmaniella, Humicola, Monilia, Monodyctis, Mortierella, Mucor, Paecilomyces, Penicillium, Phoma, Scopulariopsis, Stachybotrys, Stemphilium, Trichoderma, Trichothecium, Ulocladium, Verticillium and Wardomyces). OTA-producing moulds Aspergillus niger van Tiegham, A. ochraceus Wilhelm, Penicillium aurantiogriseum Dierckx and P. chrysogenum Thom were found as contaminants of a large number of forage samples (A. niger 22% samples, A. ochraceus 2.5% samples, P. aurantiogriseum 58% samples, P. chrysogenum 11% samples). It was found that about 33% of silage samples were contaminated with OTA (traces to 180.00 g/kg). OTA was occured in all samples (6) tested in winter period. OTA was established in 50% of hay samples at concentrations between 60.00 and 180.00 g/kg with the highest incidence in autumn. Ten of total 21 dried clover samples (47%) were contaminated with OTA (62.40 to 400.00 g/kg). OTA-positive samples were determined in autumn and winter period. Also, 3 samples of pelleted sugar beet pulp contained OTA (82.00 to 112.00 g/kg). Keywords: forage, ochratoxin-A producing moulds, ochratoxin A References: Samson, R.A., E.S. Hoekstra, J.C. Frisvad, 2004, Introduction to food- and airborne fungi, 7th ed., Centraalbureau voor Schimmelcultures Utrecht, The Netherlands OchraTestTM Instruction Manual, 1997, VICAM, L.P., Watertown, MA, USA

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P56 DEVELOPMENT AND VALIDATION OF AN IMMUNOAFFINITY BASED HPLC METHOD FOR FUMONISIN DETERMINATION IN MAIZE-BASED BABY FOOD PRODUCTS M. Solfrizzo 1 2 2 1 3 3 A. De Girolamo , D. Pereboom-de Fauw , E. Sizoo , L. Gambacorta , K. Bouten , J. Stroka , A. 1 2 Visconti and H.P. van Egmond 1Institute of Sciences of Food Production, National Research Council (ISPA-CNR), via Amendola 122/O, 70126 Bari, Italy; 2RIKILT Institute of Food Safety, P.O. Box 230, 6700 AE Wageningen, the Netherlands (previously affiliated with the National Institute for Public Health and the Environment (RIVM), PO Box 1, 3720 BA Bilthoven, The Netherlands); 3Institute for Reference Materials and Measurements (IRMM), Retieseweg 111, B-2440 Geel, Belgium.
1

Fumonisins (FB1 and FB2) are regulated in maize-based foods for infants and young children in the EU. Currently no validated methods are available for this purpose, especially taking into account the target level and the challenging matrix composition of such products. As a result, a method for the determination of FB1 and FB2 in different commercial maize-based products for infants and young children was developed. The method uses extraction at 55C with an acidic mixture of methanolacetonitrile-phosphate/citrate buffer, clean-up through immunoaffinity column and fumonisins determination by high performance liquid chromatography with automated pre-column derivatization with o-phthaldialdehyde reagent. The limit of quantification of the method (signal to noise ratio of 6) was 2.8 g/kg for FB1 and 2.2 g/kg for FB2. A limited validation study involving 3 laboratories showed good results in terms of recovery, repeatability and reproducibility values which prompted us to fully validate the method with an interlaboratory study involving 12 laboratories that analysed 5 blind duplicate pairs of baby food test materials. Mean concentrations in the test materials ranged from 89.1 to 384.4 g/kg for FB1 and from 22.5 to 73.6 g/kg for FB2. Relative standard deviations for within-laboratory repeatability (RSDr) ranged from 6.8 to 23.5% for FB1 and from 7.6 to 22.9% for FB2. Relative standard deviations for between-laboratory reproducibility (RSDR) ranged from 15.4 to 26.2% for FB1 and from 21.6 to 36.3% for FB2. Mean FB1 recoveries from test materials spiked at 100 and 250 g/kg were 89 and 96%, respectively; for FB2 spiked at 25 and 62.5 g/kg recoveries were 90 and 85%, respectively. HorRat values ranged from 0.8 to 1.2 for FB1 and from 0.9 to 1.4 for FB2 when calculated according to the Horwitz equation, and from 0.8 to 1.2 for FB1 and from 1.0 to 1.7 for FB2 when calculated according to the Thompson equation. These results were all within the acceptability criteria reported in the EU Regulation n. 401/2006 for fumonisins determination.

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P57 TYPE AND CONCENTRATION OF COAGULANT EFFECT TEXTURE SENSATION OF COAGULATED-BASED PRODUCT D. Syah 1 3 R. Fahmi , , D. Supriatna 1 Southeast Asian Food and Agriculture Science and Technology (SEAFAST) 2 Department of Food Science and Technology , Bogor Agricultural University - Indonesia 3 Agroindustrial Research Center, Ministry of Industry - Indonesia
1,2

Coagulated based product play important role in many processed food. Famous example of above product type are tofu and cheese made from soybean and milk, respectively. Soycurd is a coagulated soy protein product which is formed through additioning coagulating-agent into soymilk. Formation of curd structure is an important step that affect tofus texture and determines consumers acceptibility of product The important characteristics of tofu-based products is texture sensation raised during chewing process. This research aims to study the contribution of type and concentration of coagulant in creating optimal texture sensation. We expect that texture sensation can be understood through fraction of protein coagulated according to scheme of Osborne. To answer above objective coagulation experiment using different type and concentration of coagulant concentration has been performed. Calcium sulfate and acetic acid at three concentration (0.015, 0.030 and 0.045 N) were used to formed soycurd. Curds protein separated based on its solubility, following Osborne fractionation. Proportion of each fraction was then determined and will be correlated to tecture sensation as well as texture characteristics according TAXT. The result showed that type of coagulant affected significantly (p<0.05) to the portion of fraction that obtained. The use of calcium sulfate as coagulant gave lower proportion of globulin, prolamin and glutelin than acetic acid coagulant. Otherwise, albumin showed higher portion if calcium sulfate was used. Proportion of globulin and prolamin in curd increased significantly (p<0.05) when higher level of coagulant concentration was used, in the other hand albumin proportion occured contrary. Mathematicaly, relationship between hardness and portion of Osbornes protein fraction can be explicited as Y = 401.306 -178.043 X1 +270.204 X2 -298.194 X3 +5.415 X4, where Y, X1, X2, X3, and X4 are curds hardness, albumin portion, globulin portion, prolamin portion, and gluelin portion respectively. Globulin related positively to the hardness of curd, higher portion of globulin resulting in firmer matrix of curd. Keyword : Curd, Osborne fraction, coagulant, hardness

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P58 TRADITIONAL FERMENTED SAUSAGES: MICROBIOLOGICAL RISKS AND POTENTIAL SOLUTIONS K. T. M. Tran a b b a P. J. Coloe , M. T. Le , A. K. To , B. K. May a School of Applied Sciences, RMIT University, 124 La Trobe St, Melbourne Vic 3001 Australia b School of Biotechnology and Food Technology, Hanoi University of Technology, 1 Dai Co Viet St., Hanoi, Vietnam
a,b

Nem chua is a traditional Vietnamese uncooked fermented meat product that has played a very important part in Vietnamese eating culture and tradition for generations. However the current process of manufacture, which is based on spontaneous fermentation and back-sloping, is causing increased safety concerns. This study was aimed to investigate the microbiological quality of retail nem chua in Vietnam and isolate lactic acid bacteria (LAB) from nem chua that exhibit strong acidification and bacteriocin production for potential use as protective starter culture for nem chua fermentation. Ninety nem chua samples that were available for retail sale in Vietnam over a period of thirteen months (June 2007 to July 2008) were analysed for total viable counts (TVC), LAB, Enterobacteriaceae, Escherichia coli and coagulase-positive Staphylococci using Australian Standards methods. For LAB isolation from nem chua, strong acidifying LAB were isolated on De o Man, Rogosa and Sharpe (MRS) agar with added CaCO3 (5g/L) and incubated anaerobically (30 C, o 2-3 days). LAB with potential bacteriocin-like activity were isolated on MRS agar (30 C/24 h), overlayed with Lactobacillus plantarum JCM1149 seeded in MRS semisolid agar. Antimicrobial ability of the isolates was examined by well diffusion assay. The selected LAB were identified using API 50CH kit and 16s rRNA sequencing, and differentiated to strain level using repetitive PCR and pulse field gel electrophoresis. Their in situ antagonistic ability was evaluated towards a composite mixture of E. coli in challenge studies. The outcome of the study on microbiological quality of nem chua revealed that 93% of the samples tested were classified as unacceptable based on the Australia New Zealand Food Standards Code for all comminuted fermented meat which has not been cooked during the production process. A particular concern was the contamination with Enterobacteriaceae including E. coli and coagulasepositive Staphylococci, with 93.3%, 63.3% and 73.3% of nem chua classified as unacceptable respectively. E. coli O157:H7 was also detected in nem chua but was confirmed to be non-toxigenic by multiplex-PCR. From the native LAB population in nem chua, two strains of LAB with the strongest production of acid and antimicrobial agent were isolated and identified. These two starter cultures were successfully used to control E. coli in the challenge studies with both strains capable of reducing E. coli by more than 4 logs. The successful isolation and identification of two new strains of LAB will have enormous benefit as potential starter cultures for commercial production of nem chua with export potential to the Asia Pacific region. Key words: traditional fermented meat, microbiological quality, lactic acid bacteria, protective starter culture, strong acidifying, bacteriocin, challenge study

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P59 QUALITY AND SAFETY OF FOOD IN MASS CATERING IN POLISH HOSPITALS - THE ASSESSMENT OF REALIZATION OF HYGIENIC STANDARDS AND HACCP SYSTEM IMPLEMENTANTION H.Turlejska M. Rams-witoniowska, E. Konecka-Matyjek National Food and Nutrition Institute, Department of Food Safety, Poland

Introduction: The quality and safety of food in mass catering have a big impact on health and safety of patients in hospitals. According to the Polish and UE food law, all food operators, including mass catering in hospitals, have a duty to fulfill all of hygienic and sanitary requirements, realize GHP/GMP principles and implement the HACCP system. The aim of the study: The aim of the study was to assess the hygiene including GHP, GMP and HACCP implementation in mass catering in hospitals. Material and methods: The study was carried out in collaboration with Chief Sanitary Inspectorate, and the Highest Control Chamber. In the study the results from direct control activity of Sanitary Inspection within the years of 2003 2008 and Highest Control Chamber were included. There were 900 hospitals assessed within official control activity as well as directly 12 hospitals and by questionnaires sending 125 hospitals within Highest Control Chamber control conducted in 2008. Results and discussion: The results show that less than 1 % of establishments dont fulfill legal requirements. Almost 95% of hospital canteens realized all GHP/GMP principles and kept sufficient documentation in this area. The results from controls provided in collaboration with Highest Control Chamber show some discrepancies in the area of food safety management, especially in the frame of technical conditions, meals processing and distribution. There were also observed some negligence in knowledge and qualifications of the main canteens` staff. The degree of HACCP system implementation was not satisfactory. It was assessed that only 43% of hospital kitchens had HACCP implemented in 2008. This result was better than in the previous year, where only 27% of establishments declared its implementation. The results show that a lot of hospital canteens are at the first stage of HACCP implementation and their system doesnt fully work at this moment. Conclusions: The analysis of the results from sanitary and hygienic control activity show that after 2006 the sanitary conditions in hospital canteens were improved Hospital canteens are not satisfactory involved in HACCP system implementation .Some of hospital canteens have a lot of difficulties in HACCP implementation system and they need practical expert consultations and support in this subject. Key words: hospitals, mass catering, nutrition, food safety, food quality, GHP/GMP, HACCP system, food law requirements. Basic references: The information on the inspection results on nutrition and cleaning and disinfection in hospitals, Warsaw, Highest Control Chamber, 2009 The results from the official food control inspection from years of 2003 2008. Chief Sanitary Inspectorate unpublished data.

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P60 FOOD ALLERGENS AND HACCP: HAZARDS INDUCED AND ADDITIONAL CONTROLS C. Tzia T. Tsirka, V. Giannou, D. Lebesi, V. Polychniatou, C. Chranioti & P. Sfakianakis Laboratory of Food Chemistry and Technology, School of Chemical Engineering, NTUA, Polytechnioupoli, Zografou, 15780 Athens, Greece Tel: (30210) 7723165, Fax: (30210) 7723163, E-mail: tzia@chemeng.ntua.gr

Food allergens are hazards of high importance for the food industry as they can cause adverse reactions to a special part of consumers (allergic to specific food or substance of food). Food allergy constitutes a category of non toxic adverse reactions to food occurring via antibodies which are considered responsible for a variety of gastrointenstinal, cutaneous and respiratory disorders. IgE and non IgE-mediated food allergies are referred; IgE-mediated food allergies are the most important where the antibody that takes part in this adverse reaction as an intermediate is the immunoglobulin E. Food allergy appears in a respectable percentage of people around the world. Specific foods or food groups are thought to cause the majority of food allergies on a worldwide basis i.e. milk, eggs, fish and crustacean, soybean, peanuts, wheat etc. The components of these foods that elicit food allergies may be naturally occurring proteins. Food allergies can be also caused by proteins of genetically modified food products. A systematic approach to assessment of the allergenic properties of novel food proteins testing is based upon similarities with known protein allergens and consideration of protein stability. Animal models that mimic the antibody response or even the symptoms upon a challenge reaction as observed in humans are also used for the risk assessment of food allergens. FAO/WHO recommends the use of a decision-tree for the assessment of the allergic potential of novel food proteins. In European countries, allergens should be indicated in food labels of pre-packed and drinks according to Reg. 2000/13/EC, 2003/89/EC. A food industry may produce food products that naturally contain allergenic components (A), food products that regularly do not contain allergens but can receive allergic potential inside the industry throughout the processing (B1) or free allergens products (B2) (i.e. gluten -free bread). In this study the food allergens hazards induced in typical cases (A, B1, B2) of food products in a food industry, the hazard analysis and the additionally required controls, analyses and actions for the HACCP system related to the CCPs and PRPs from raw material receipt to food processing, packaging and labelling are presented. KEYWORDS: food allergens, HACCP, food hazard, risk assessment REFERENCES Koppelman, S.J., Hefle, S.L. (2006). Detecting allergens in food, CRC Press, Cambridge, England, Ch.2. Crevel, R.W.R., Briggs, D., Hefle, S.L., Knulst, A.C., Taylor, S.L. (2007). Hazard characterisation in food allergen risk assessment: The application of statistical approaches and the use of clinical data, Food and Chemical Toxicology, 45, 691-701. Van Hengel, A.J. (2007). Declaration of allergens on the label of food products purchased on the European market, Trends in Food Science & Technology, 18, 96-100.

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P61 VERIFICATION OF THE IDENTITY OF FATS AND OILS S. van Ruth W. Akkermans, M. Rozijn, A. Koot, H. van der Kamp, A. Tres, M. Alewijn RIKILT-Institute of Food Safety, Wageningen University and Research Center, Wageningen, the Netherlands

Product authenticity and authentication are emerging topics within the food sector. It is a major concern not only for consumers, but also for producers and distributors. This topic is important from a fair trade perspective, for consumer confidence reasons, but also from a food safety point of view (e.g. melamine). Food adulteration has been practiced forever, but has become more sophisticated in the recent past. Foods or ingredients most likely to be targets for adulteration include those which are of high-value and which undergo a number of processing steps before they appear on the market. Fats and oils are generally high-value products and can therefore be considered targets. Traditional analytical strategies to uncover adulteration and guarantee quality have relied on wet chemistry to determine the amount of a marker compound or compounds in a suspect material and subsequent univariate comparison of the value(s) obtained with those established for equivalent material of known provenance. This approach suffers from a number of disadvantages, namely, the ever-increasing range of analytes which must be included in any test procedure and the limited knowledge of the range of each constituent in normal lots of the substance. Therefore, there is a continuing demand for new, rapid, non-destructive, cost-efficient methods for direct quality measurements in fats and oils in order to verify their identity. The presentation will involve an overview of traditional and novel strategies in the area of fat and oil authentication, with emphasis on the latter. A study on the verification of the identity of fats and oils with application of GC profiling and PTR-MS profiling combined with chemometrics will be presented. Sample material involves animal fats and vegetable oils. Estimated and validated classification models will be shown which allow prediction of the identity of the various fats/oils with reasonable success.

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P62 A WORLD TRADE MARKET DILEMMA OF RAPESEED (SEED-OIL); A COUNTERVAILING RISK OF A SYNERGISTIC INTERACTION BETWEEN AFLATOXIN B1 AND HEPATITIS B/C, LEADS TO A SIGNIFICANT HEALTH RISK AND FOOD-BORNE ILLNESS, IS STUDYING VS. THE ECONOMIC IMPACT IN CHINA AND EU. M. Vered 2 2 P. LI and X. Ding 1.VTAG Israel 2.Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Xudong 2Rd, Wuchang, Wuhan, P.R. China.
1

A synergistic interaction between carcinogenic effects of aflatoxin B1 (AFB1) and chronic hepatitis B virus (HBV) infection, offers a plausible explanation for the very high incidence of the tumor and the young age of the patients in China, or Africa. Both chronic HBV infection and repeated exposure to AFB1 are known to be major risk in the multifactor etiology of human hepatocellular carcinoma HCC (1). The highest incidences of HCC among the youngest patients with this tumor are found mostly in China, as hyper endemic for HBV infection has a high rate of dietary exposure to the fungal toxin, AFB1. Even so, China with high hepatitis prevalence would likely to be the larger export of its best rapeseed commodities and keep worst at home. This choice would increase health risk and public is expected for strengthening the local standard to protect food safety. On the sometime EU (as import region) maintain on harder AFB1 standards, but dose it really to protect the public and provide food safety? Unlike EU, China as export region is very much concerning for future export. The risk is related to the healthy threat from mycotoxin that increasing dramatically the possibility for liver cancer caused by HBV. That "meeting point" between mycotoxin and hepatitis is both an ecological disaster of tropic areas, and economical dilemma to policy makers in EU and China. The presence of mycotoxin in rapeseeds or rapeseed meal is yet within the limit admitted by the acting EU regulation (< 50 ppb) for aflatoxin and 5 ppm for deoxinivalenol and zearlenone (2), nevertheless, rapeseed will be soon a major source of production seed-oil, and it is expected to have strong economical impact in the next decade on seed-oil world marketing. By framing the dilemma as an economic vie health risk this project is attempting to move policy decisions and international regulations toward more reasonable balance that can aid decision makers in creating safe and feasible mycotoxin standards. The combination of AFB1 with HBV, is a synergistic factor, that raising more than ~60-fold the risk of liver cancer and the objective of this study is to gain data on health risk and economic impact of AFB1 (3). More specifically, the goals are to identify the nations those which would suffer from heavily impacted by tighter mycotoxin standards and address riskrisk tradeoffs between health benefits an d economic losses from compliance with those regulations. Risk assessment will help to develop more effective HACCP seed-oil plans in China and else-where. For the future, it will help develop HACCP for rapeseed's seed-oil production. In order to select the best option, a pragmatic economical model that would be taken in consideration to different AFB1 standards, and placing them on a mathematic equation, is Monte Carlo simulation model. Under this uncertainty the model will provide an answer (risks) on level of human illness vis-a-vis the economical impact. Keywords: rapeseed, seed-oil, mycotoxin, hepatitis B virus, risk assessment, Monte Carlo

Reference: (1) M.C. Kew 2003 (2) C. Tabuc and G. Stefan Arc. Zootechnica vol 8, 1-6 2005 (3) Groopman, J.D., Kensler, T.W. and Wild, C.P 2008. Protective Intervention to Prevent aflatoxin induced carcinogenesis in developing countries. Annual Review of Public Health 29:187-203

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P63 THE OCCURRENCE OF FALSE POSITIVE SAMPLES IN DETERMINATION OF OTA IN HERBAL TEA, SPICES AND DIETETIC PRODUCTS G. Vukovi V. Palibrk, M. Stankovi, M Tadi Institute of Public Health, Despota Stefana 54A, Belgrade, Serbia

The occurrence of ochratoxin A in common herbal tea, dietetic products and spices was examined. Composition of mycopopulation and mycotoxins were analysed in 57 samples of dietetic products, herbal tea and spices or their mixtures. The most dominant fungi recorded were Aspergillus flavus, Aspergillus niger, Penicillium sp., Fusarium verticillioides, and Fusarium sp., already well known producers of the mycotoxins. The method used involves the high-performance liquid chromatography (HPLC) with the fluorescence detection (FLD). The ochratoxin A was extracted using a mixture of methanol and water isolated and concentrated by means of immunoaffinity column chromatography. OTA was separated using the RP-18 column and detected by HPLC/FLD. The method permits the detection of 0.25 ppb OTA and recovery of 85% (with relative standard deviation less than 5%). Of the 57 samples, 27 were found to be contaminated with OTA, and, of these, 8 did not comply with EU regulation. All positive samples were also analysed by liquid chromatography/electrospry ionisation tandem mass spectrometry (LC/ESI-MS/MS) method, and 3 samples out of 27 were found to be false positive. It was interesting that these false positive samples showed high amounts of OTA (from 300 to 600 g/kg), when analysed by HPLC/FLD. Determination of OTA by immunoaffinity columns clean up and HPLC/FLD detection was shown to be unspecific for heavy matrix samples such as herbal tea, spices and dietetic products, since there were 3 false out of 27 positive samples. It was proved that use of the LC/MS/MS as confirmatory technique was necessary in analysis of such samples. Key words: mycotoxins, moulds, ochratoxin A, HPLC/FLD, LC/ESI-MS/MS, herbal tea, dietetic products, spices. References: G.Vukovic, S.Pavlovic, M.Ristic, Comparison of two sample preparation procedures for HPLC determination of ochratoxin A, Arch. Biol. Sci., Belgrade, 61(4), 639-644, 2009. C. Juan at al. Determination of ochratoxin A in maize bread samples by LC with fluorescence detection, Talanta 73 (2007) 246250 B. P.-Y. Lau at al. Quantitative determination of ochratoxin A by liquid chromatography/electrospray tandem mass spectrometry, J. Mass Spectrom. 35, 2332 (2000) Method Validation & Uncertainty Report Determination of Ochratoxin A in Cereals and Pasta using Immunoaffinity Column Clean Up and HPLC with Fluorescence Detection; Food Laboratories Division, Ontario and Nunavut Region, Health Products and Food Branch, Food Chemistry Laboratory, 2301 Midland Avenue, Scarborough, Ontario

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P64 NEW RELIABLE METHOD USING MYCOSEP 150 CLEAN UP COLUMN TO DETECT ERGOT ALKALOIDS IN AGRICULTURAL COMMODITIES E. Wanzenbck 2 1 1 1 E. Halbmayr , B. Daxner , M. Schandl , E. Pichler 1 Romer Labs Diagnostic GmbH, Technopark 1, 3430 Tulln, Austria 2 Romer Labs Division Holding GmbH, Technopark 1, 3430 Tulln, Austria
1

Ergot alkaloids are mycotoxins produced by fungi of all members of the Claviceps species, most important in terms of frequency of occurrence C. purpurea. These fungi are able to produce a wintering body, also known as sclerotium. Such structures are mainly found on rye, wheat and triticale. Sclerotia contain different classes of alkaloids, the most prominent being ergometrine, ergotamine, ergosine, ergocristine, ergocryptine and ergocornine. The amount and pattern of each alkaloid vary between fungal strains, depending on the host plant and the geographical region. Ergot alkaloids exert toxic effects in all animal species, and the most prominent symptoms are feed refusal, dizziness, insufficient nursing of suckling animals due to agalactia and even abortions. The interest for ergot alkaloids in food and feed is increasing since there are no formally validated official methods for those analytes. This work shows the results of a new reliable HPLC-FLD method using a one step clean up solid phase extraction column (MycoSep 150) after extracting the sample with acetonitrile/ammonium carbonate. The validation was carried out using Biopure dried down ergot alkaloid standards which where dissolved in acetonitrile and spiked to the samples prior to extraction. Six parallels of blank commodities as well as six replicates of three spiking levels were analyzed by different analysts. Results showed excellent recoveries of 80 % for all analytes and very sensitive limits of detection between 7 and 13 g/kg and limits of quantification between 20 and 40 g/kg for different ergot alkaloids in commodities rye and wheat. The method is reliable and easy to use for routine analysis of ergotamine, ergosine, ergocristine, ergocryptine and ergocornine. Keywords: Ergot alkaloids, clelan up column, HPLC method References: Romer Labs Application Brief Rapid Quantitation of the 6 major Ergot Alkaloids in Grian by HPLC FLD Kromidas S., Validierung in der Analytik, ISBN 3-527-28748-5 Commission Regulation (EC) No 401/2006 Laying down the methods of sampling and analysis for the official control of the levels of mycotoxins in foodstuffs

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P65 DETECTION OF TOXIC FRAGMENTS FROM GLUTEN USING A NEW MONOCLONAL ANTIBODY-BASED TEST E. Wanzenbck 2 3 3 4 E. Halbmayr , R. Fielder , A. Rogers , M. Zheng 1 Romer Labs Diagnostic GmbH, Technopark 1, 3430 Tulln, Austria 2 Romer Labs Division Holding GmbH, Technopark 1, 3430 Tulln, Austria 3 Romer Labs UK ltd, North Wales Business Park, Building Unit 5325 Abergele, LL22 8LJ North Wales, United Kingdom 4 Romer Labs Singapore Pte. Ltd., 3791 Jalan Bukit Merah #08-08, e-Centre@redhill building, Singapore, 159471 Celiac disease (CD) is an immune-mediated enteropathy caused by the ingestion of gluten, a protein fraction found in certain cereals. Immunotoxic gluten peptides that are resistant to degradation of digestive enzymes appear to trigger celiac syndrome. Celiac disease occurs in genetically predisposed persons and leads to the destruction of the microscopic finger-like projections of the small intestine, called villi. The disease is triggered by the ingestion of peptides from wheat, barley, rye, and in some cases oats. It currently affects roughly 1% of the worlds population, primarily adults. This work is showing the results of a new monoclonal antibody that specifically recognises the pathogenic fragment of the gliadin protein present in gluten. This fragment is called 33-mer and triggers the auto-immune reaction in celiac patients. Homologues of this peptide were found in every food grain (except oats) that is toxic to CD patients, but were absent in all nontoxic food grains. The antibody was specially developed to determine the toxic fractions present in gluten. When food and drinks are hydrolysed or heat processed, the gliadin protein is degraded in small fragments which are difficult to detect with classic antibodies because their epitopes may be destroyed. The newly developed monoclonal antibody can reliably detect toxic fragments of gluten in hydrolysed food. The outstanding advantage of this new antibody is the possibility to detect the actual toxic fragment of gluten with a very high sensitivity. Due to current Codex Alimentarius recommendations and Commission Regulation (EC) No 41/2009, food can be labelled gluten free when containing less than 20 mg/kg gluten. A lateral flow test kit using this monoclonal antibody was developed to detect the toxic fractions of gluten from wheat and other cereals such as barley, rye and, with a much lower level of sensitivity, oats. The semi-quantitative immunochromatographic strip test is based on sandwich format. The reagents for the test and control line are immobilized on a nitrocellulose membrane. Toxic gluten fragments in the sample extract react with anti-gliadin 33-mer monoclonal antibody, coupled to coloured microspheres, which is dried on the strip showing a visible line when binding to the anti-gliadin 33-mer monoclonal antibodies on the test line. The mix of conjugate moves through the membrane to the control line where anti-species specific antibodies used for verifying the correct test performance are sprayed. Keywords: gluten, G12 monoclonal antibody, immunochromatographic strip test, 33-mer
References: Shan L, Molbergv , Parrot I, Hausch F, Filiz F, Gray GM, Sollid LM, Khosla C. Structural basis for gluten intolerance in celiac sprue. Science. 2002 Sep 27;297(5590):2275-9. Morn B, Bethune MT, Comino I, Manyani H, Ferragud M, Lpez MC, Cebolla A, Khosla C, Sousa C. Toward the assessment of food toxicity for celiac patients: characterization of monoclonal antibodies to a main immunogenic gluten peptide. PLoS ONE. 2008a May 28;3(5):e2294. Morn B, Cebolla A, Manyani H, Alvarez-Maqueda M, Megas M, Thomas Mdel C, Lpez MC, Sousa C. Sensitive detection of cereal fractions that are toxic to celiac disease patients by using monoclonal antibodies to a main immunogenic wheat peptide. Am J Clin Nutr. 2008b Feb;87(2):405-14.
1

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P66 COMPARATIVE STUDY ON SAFETY ASSESSMENT FOR FLAVOURINGS OF CAC, UNITED STATES, EUROPEAN UNION AND CHINA Xiao-fen Yao Jin-yao Chen, Li-shi Zhang Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University, China

Background/Objectives: Flavourings have been widely applied in food industry. In the recent years, international trade of flavourings has flourished, especially in Asia-Pacific region. Though the current consumption of flavourings per capita in China is relatively low, the demands of flavourings in China might grow at the fastest speed in the next decade. Furthermore, there are abundant natural flavourings resource and great market potential in China. But there exist many deficiencies in safety assessment in China, which causes it is not enough to meet the need of food safety and market development. On the other hand, the safety assessment of flavourings has its own particularity comparing to other categories of food additives. In this study, the safety assessment systems in China, the United States, the European Union and the CAC were compared, and based on it some suggestions to improve safety assessment were put forward. Methods: Current and Valid regulations and methods on safety assessment of flavourings in the CAC, the United States, the European Union and China were collected, compared and analyzed. Results: There are specific agencies for safety assessment of flavourings, methods for evaluating exposure, and procedures for safety assessment in the CAC, the United States, the European Union and China. Considering the specific agencies, in the CAC it is Joint FAO/WHO Expert Committee on Food Additives (JECFA), Flavour Extract Manufacturers Association (FEMA) is in charge in the United States, in the European Union it is the European Scientific Committee for Food (SCF) of European Food Safety Authority (EFSA) .However, China lacks uniform safety assessment agencies, and they are dispersed in provinces, municipalities and autonomous regions. For exposure evaluation, CAC mainly adopts maximized survey-derived intake (MSDI) and single-portion exposure technique (SPET), FEMA mainly uses the possible average daily intake (PDAI) and per capita intake (PCI), the European Union applies MSDI, SPET, theoretical added maximum daily intake (TAMDI) and modify theoretical added maximum daily intake (mTAMDI), while no specific evaluation methods at present in China. SPET is a new method proposed by JECFA in 2007, and PCI 10 is similar to the MSDI. In the assessment procedures, CAC, United States and the European Union mainly adopt Flavouring Groups Evaluations (FGE), referring to the JECFA s decision tree of assessment procedures, and the assessments in China are conducted according to GB15193.1-2003, which provides guidelines about toxicological assessment procedures of flavourings. Additionally, although the assessment procedure of JECFA is a new and advanced method, it also holds a major deficiency, i.e. flavourings are only defined from the chemical point of view in JECFA, and at present only assessment procedures of chemicals were put forward. The future focus of work is to include some natural flavourings, especially to study the new evaluation method for some natural complex mixtures. Conclusions/Suggestions: The defects and suggestions of the system of safety assessment for flavourings in China are included: (1) It is difficult to ensure accuracy of results of safety evaluation with lacking of a systematic and uniform safety assessment agencies. (2) Methods for exposure evaluation have not been determined. Considering our country's specific national conditions, MSDI is more suitable, for it is simple, convenient, rapid, economical, and the evaluation results are more accurate. (3) Lacking of proper safety assessment procedure makes it difficult to meet the needs of food safety and international trade. The effective evaluation system should be established and data should be updated timely to re-evaluation of approved flavorings, in order to ensure their safety. (4) A wide-range, easily accessible toxicity information database for safety assessment of flavourings has not been established in China. (5) In the collection of exposure data, some experiences from developed countries could be taken in, e.g. establishing web site information collecting system to gather data as soon as possible and to ensure the accuracy of evaluation results. Key words: flavourings, safety assessment, exposure evaluation

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References: Chen Jun- Shi. The Role of Science in Codex Standards[J]. Biomedical and Environmental Sciences,2001,14(1- 2):145- 148. Schrankel KR. Safety evaluation of food flavorings [J]. Toxicology, 2004,198. John B. Hallagan, Richard L.Hall. FEMA GRAS-A GRAS Assessment Program for Flavor Ingredients. Regulatory Toxicology and Pharmacology, 21:422-430(1995). EFSA-CEF - Food contact materials, enzymes, flavourings and processing aids http://www.efsa.europa.eu/EFSA/ScientificPanels/efsa_locale-1178620753812_CEF.htm Evaluation of food additives and contaminants [R] . Sixty-eighth report of the joint FAO/WHO Expert Committee on Food Additives, June 2007 19-28. Robert L. Smith, Samuel M. Cohen, John Doull. Criteria for the safety evaluation of flavoring substances The Expert Panel of the Flavor and Extract Manufacturers Association [J]. Food and Chemical Toxicology,2005,43: 1141-1177. Xu YiCao Yi, Jin Qi-Zhang. The safety concern and regulation & standards for flavorings [J].China Food Additives, 2009,2:49-54. IOFI AND CODEX ALIMENTARIUS http://www.iofi.org/Iofi/English/Home/Flavor-Safety/IOFI-andCodex-Alimentarius-Commission/page.aspx/35.

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P67 PRIMARY STUDY ON THE LOOP-MEDIATED ISOTHERMAL AMPLIFICATION METHOD FOR THE RAPID DETECTION OF STAPHYLOCOCCUS AUREUS Zhimei Xie 2 3 4 Heng Chen , Yi Teng , Xuejun Fan , Xiaofang Pei* 1 West China School of Public Health, Sichuan University, Chengdu, China; 2 Chengdu CDC; 3 Long Quan Yi CDC, Chengdu; 4 Sichuan Entry-Exit Inspection and Quarantine Bureau [Abstract] Staphylococcus aureus is responsible for most food poisoning through the production of enterotoxin. A rapid and simple detection method is of great importance for field use especially in developing countries. So Loop-Mediated isothermal amplification (LAMP) technique, which is more sensitive and less time-consuming than PCR and needs no thermal cycler, becomes a potential candidate. Objective To establish the Loop-Mediated Isothermal Amplification Method for the rapid detection of Staphylococcus aureus for field use. Methods Since the enterotoxin positive Staphylococcus aureus is parallel with the production of thermostable nuclease, we selected NUC gene as target gene, and designed specific LAMP primers which can identify several regions of target gene. Through altering the quantity of key reagent (buffer, MgSO4, Betaine, dNTP, Bst DNA, four primers), the temperature and the time of reaction to optimize amplification condition, we primarily established the LAMP method for rapid detection of Staphylococcus aureus. Results NUC gene of Staphylococcus aureus which was selected as the target gene of LAMP can be successfully employed and the four specific primers F3, B3, FIP, BIPdesigned for this target gene had satisfied specificity. And through optimizing the influential factors, such as Mg , dNTP, Betaine, F3 and B3, FIP and BIP, Bst DNA polymerase, the LAMP detection of Staphylococcus aureus we developed can be completed within 70 min at 60in water-bath. Conclusion A Loop-Mediated Isothermal Amplification Method was established for the rapid detection of Staphylococcus aureus. This will be helpful for the further Staphylococcus aureus detection in field workfor the rapid screen of Staphylococcus aureus in food and will be a potential useful food safety supervision tool in developing countries. [Key words] Staphylococcus aureus; Loop-Mediated isothermal amplification (LAMP) technique [References] Notomi T, Okayama H, Masubuchi H, et al. Loop-mediated isothermal amplification of DNA [J]. Nucleic Acids Res, 2000, 28(12):63. Mori Y, Nagamine K, Tomita N, et al. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun, 2001, 289: 150-154. ----------------------------* Corresponding author. Fax: +86 28 85501275 (fax); E-mail address: xxpeiscu@163.com Acknowledge: This work was partially supported by MoniQA (FOOD-CT-2006-036337) and scientific research project of General Administratioin of Quality Supervision, Inspection and Quarantine of the Peoples Republic of China (NO. J2005J0115)
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MoniQA Coordinator:

Dr. Roland Ernest Poms, ICC International Association for Cereal Science and Technology, Vienna, Austria Marcella Gross, ICC International Association for Cereal Science and Technology, Vienna, Austria Daniel Spichtinger, RTD Services, Vienna, Austria Sin Astley, IFR, United Kingdom

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