Chemical Identification of Contaminants in Paper and Board Food Contact Materials
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Downloaded from orbit.dtu.dk on: Dec 21, 2017 Chemical identification of contaminants in paper and board food contact materials Bengtström, Linda; Petersen, Jens Højslev; Granby, Kit; Trier, Xenia Publication date: 2014 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Bengtström, L., Petersen, J. H., Granby, K., & Trier, X. (2014). Chemical identification of contaminants in paper and board food contact materials. Kgs. Lyngby: Technical University of Denmark (DTU). 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Chemical identification of contaminants in paper and board food contact materials Linda Bengtström PhD Thesis 2014 Chemical identification of contaminants in paper and board food contact materials PhD Thesis Linda Bengtström Division of Food Chemistry National Food Institute Technical University of Denmark December 2014 Thesis title Safety of food packaging materials - Development of a bioassay guided strategy for the identification of contaminants in paper and board Author and place of employment Linda Bengtström Division of Division of Food Chemistry National Food Institute Technical University of Denmark Supervisors Senior researcher Jens Højslev Petersen Division of Division of Food Chemistry National Food Institute Technical University of Denmark Deputy Head of Division, Senior researcher Kit Granby Division of Division of Food Chemistry National Food Institute Technical University of Denmark Researcher Xenia Trier Division of Division of Food Chemistry National Food Institute Technical University of Denmark Evaluation committee Team leader and senior scientist, Dr. Emma Bradley, The Food and Environmental Research Agency, UK Ass. Professor Bo Svensmark, Department of Plant and Environmental Sciences, Copenhagen University, DK Senior scientist, Dr. Julie Boberg, National Food Institute, Technical University of Denmark, DK Cover photo Fractions from a recycled pizza box, photographer Linda Bengtström Copyright Reproduction of this publication in whole or in part must include the customary bibliographic citation, including author attribution, report title, etc. Published by Technical University of Denmark, Ledelse og Administration, Anker Engelunds Vej 1, Bygning 101 A, 2800 Kgs. Lyngby, 2014 Funding The Ministry of Food, Agriculture, and Fisheries of Denmark and the Danish Veterinary and Food Administration Chemistry is like cooking, just don’t lick the spoon. -Unknown i Preface The practical work for this PhD has been conducted at The National Food Institute, Technical University of Denmark. In addition, practical work was also performed at the Food and Environmental Research Agency (FERA), UK under the supervision of Dr. Malcolm Driffield. The work presented in this thesis was funded by the Danish Government (Fødevareforlig II). The stay at FERA was generously funded by a grant from the NordFluor network. Acknowledgement There are so many people I would like to thank for their involvement in my PhD project. First of all, I would like to thank my supervisors Jens Højslev Petersen, Kit Granby and Xenia Trier for your guidance and support throughout the project. I want to thank all the colleagues involved in the Cocktail V project for fruitful discussions and for establishing a creative and innovative environment absolutely essential for the success of this interdisciplinary study. Also, thanks to the laboratory technicians Annie Foverskov, Anni Helleskov, Lone Falk Hertz and to laboratory engineer Lisbeth Krüger Jensen. Without your guidance and skills I would have struggled far more in the laboratory. Thanks to all the co-workers at Dept. K (Food Chemistry) for giving me such a happy time during my years at the department as well as being helpful. Thank you to my office mates; Kasper, Anne-Mette and Line for all the good times and for sharing my belief that no question is too silly not to be googled. A shout out for all my PhD peers in the ULF network at the National Food Institute. Thank you for all your cromulent support and for all the laughs. As part of my PhD project I got the opportunity to visit the Food and Environmental Research Agency (FERA) in York, UK. I had the pleasure of being supervised by Dr. Malcolm Driffield, whom together with his research group introduced me to new techniques, instrumentation and strategies for identification. Everyone was very welcoming and helpful throughout my stay in York. Thanks to Michael Dickinson and Antony Lloyd for helping me acquire and interpret data. Likewise, thanks to Lisa Parker Gomm for being such a sweet landlord in the lovely city of York. Finally, I want to thank my friends and la familia. A special thank you goes to my mother Monica Bengtström, my father Hans Bengtström and my little sister Malin Bengtström. Lastly, I want to thank Jill, my Poison. København/Malmö (København M) December 2014 Linda Bengtström PhD student ii Summary Paper and board are used for a variety of food contact materials, such as baking paper, microwave popcorn bags and packaging for cereals as well as fast foods. Despite this extensive use, there are currently large data gaps about the chemical composition of different paper and board food contact materials and the toxicological effects of these compounds. The aim of this study was to develop a rationalised interdisciplinary strategy for the screening and identification of compounds with potential adverse health effects in paper and board materials. The first step in the proposed strategy was to develop a comprehensive extraction process that is compatible with both chemical and toxicological analyses. For this purpose, a purge-and-trap method was developed for the collection of small volatile organic compounds; in addition semi- and non- volatile compounds were extracted by a boiling ethanol reflux system. After an initial in vitro screening of 20 different paper and board samples for endocrine disruptive effects, mutagenicity and effects on metabolism of foreign compounds, five samples with adverse effects were selected for fractionation. The fractions were tested in cell assays in a second screening. The fractionation was used to reduce the number of compounds to be identified as well as the matrix effect. Next, the fractions were analysed by gas chromatography and liquid chromatography coupled to high resolution mass spectrometry. These two techniques were designed to be as complimentary as possible and by them in combination increased the possibility to identify compounds with potential adverse health effects. Several steps in the tentative identification by gas chromatography can be automated, due to the standardisation of this technique that enables searches in vast mass spectral libraries. Such libraries are missing for liquid chromatography, and a large part of the tentative identification must be performed manually. To facilitate the tentative identification by liquid chromatography, an accurate mass database containing approximately 2100 entries of compounds with reported use in paper and board was built. The results from this study indicate that both isotope ratio and hits in the accurate mass database greatly increases the possibility of a correct tentative identification. After lists of tentatively identified compound had been produced for a certain toxicological assay, compound were selected for further testing based on previously reported effects, structural similarities to known ligands, and availability of analytical standards for identified compounds. Any positive annotation through databases should be regarded as tentative, and therefore analytical standards were used to confirm the identification. After confirmation, equivalence factors for the initially observed toxicological effect and from all the confirmed compounds tested in the same toxicological assay were calculated. The initially observed effects on the metabolism of xenobiotics could to a minor extent, though not fully, be attributed to dyes used in printing inks. In addition, it was concluded that the endocrine disruptive effects could largely be explained by monomers and plasticisers present in a recycled fibre sample and by sizing agents in virgin fibres. iii Resume’ (summary in Danish) Papir og pap anvendes til en række forskellige materialer i kontakt med fødevarer, såsom bagepapir, mikrobølge-popcorns poser og emballager til morgenmadsprodukter og fastfood. Trods den omfattende anvendelse, mangler der viden både om den kemiske sammensætning af forskellige fødevarekontaktmaterialer af pap og papir og de toksikologiske effekter af disse forbindelser. Formålet med dette PhD-studie var at udvikle en rationel interdisciplinær strategi til screening og identifikation af kemiske stoffer i materialer af pap og papir med potentielt sundhedsskadelige effekter. Det første skridt