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Analytical Development and Traceability in Food Chemistry

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Analytical Development and Traceability in Food Chemistry Analytical development and traceability in food chemistry. Examples of application to Swiss and foreign Emmental cheese A THESIS Submitted to the UNIVERSITY OF NEUCHATEL For the degree of DOCTOR OF TECHNICAL SCIENCES Presented by LAURENT PILLONEL Dipl. Chem. ETH Born on 19th June, 1975 Citizen of Seiry (FR) Accepted on the recommendation of Prof Dr R. Tabacchi Prof Dr T. Turlings Dr J.O. Bosset Dr J.F. Chamba Dr J.L. Luisier Dr E. Pfammatter Neuchâtel, February 2004 Développement analytique et traçabilité en chimie alimentaire. Exemples d'application à des Emmental suisses et étrangers Une thèse Soumise à l’ UNIVERSITÉ DE NEUCHÂTEL Pour le degré de DOCTEUR EN SCIENCES TECHNIQUES Présenté par LAURENT PILLONEL Dipl. Chem. ETH Né le 19 juin 1975 Originaire de Seiry (FR) Accepté sur la recommandation de Prof Dr R. Tabacchi Prof Dr T. Turlings Dr J.O. Bosset Dr J.F. Chamba Dr J.L. Luisier Dr E. Pfammatter Neuchâtel, février 2004 Acknowledgements My warmest thanks go to Dr Jacques-Olivier Bosset (FAM, Berne) who gave me the idea for the project. His patience and goodwill as well as his huge knowledge of cheese and food chemistry were of great help throughout the thesis. I would like to express my sincere gratitude to Prof Dr Raffaele Tabacchi (Université de Neuchâtel) for giving me the opportunity to carry out this thesis and for the confidence he had in this project. I am grateful to Dr Christian Steffen, director of the FAM, for putting the infrastructure and the knowledge of all colleagues at my disposal. I would like to thank the CTI program, the Swiss Federal Office of Public Health and Emmentaler Switzerland™ for their financial support of the project. Furthermore I would like to thank: Prof Dr Ted Turlings (Université de Neuchâtel), Dr Jean-François Chamba (ITFF, Foron/Roche, France), Dr Elmar Pfammatter (Laboratoire cantonal du Valais, Sion) and Dr Jean-Luc Luisier (HESVs Sion) for agreeing to be co-examiners of the thesis. Jean-François Chamba (ITFF), Eric Dufour (ENITA, Lempdes, France), Giuseppino Fortunato (EMPA, St. Gallen), Pascal Froidevaux (IRA, Lausanne), Alfred Jakob (Labor Spiez, Spiez), Georg Haberhauer (ARC, Seibersdorf, Germany), Stephan Hölzl (Universität München, Germany), Peter Horn (Universität München), Elmar Pfammatter (Laboratoire cantonal du Valais), Umberto Piantini (HEVs), Daniel Picques (INRA, Grignon, France), Andreas Rossmann (Isolab, Schweitenkirchen, Germany), Emmanuelle Schaller (Büchi, Flawil), who kindly offered analyses from their laboratories. A project of this scale would never have been possible without the interest and the willingness of such scientific partners. Thierry Zesiger (LDZ, Marin) for his technical support with electronic noses. Gerda Urbach and Gérard Gremaud for reviewing the manuscripts as well as Robert Sieber for correcting the German summaries. Ueli Bütikofer (FAM) and Werner Luginbühl (FAM) for their great help with the statistical exploitation of the data. All colleagues at the FAM for the interesting discussions and the kindness I found everywhere. My family and friends for their company in the alternative programs. The Swiss chocolate industry for manufacturing the most important food product a Ph.D. student could ever need. Table of contents TABLE OF CONTENTS…………………………………………………………………. I SUMMARY.......................................................................................................................... III ZUSAMMENFASSUNG…………………………………………………………………. IV RÉSUMÉ………………………………………………………………………………….. V RIASSUNTO……………………………………………………………………………… VI PREAMBLE………………………………………………………………………………. VII PART I R&D IN VOLATILE COMPOUNDS AND ELECTRONIC NOSES 1. Rapid preconcentration and enrichment techniques for the analysis of food volatiles: a review………………………………………………………………… 1 2. Long term study of volatile compounds from deep frozen, canned, processed cheeses proposed as control standards……………………………………………. 15 3. Data transferability between two MS-based electronic noses using processed cheeses and evaporated milk as reference materials……………………………… 29 4. Comparison of efficiency and stability of two preconcentration techniques (SPME and INDEx) coupled to an MS-based “Electronic Nose”………………... 32 PART II AUTHENTICATION OF EMMENTAL CHEESE: A FEASIBILITY STUDY 5. Analytical methods for the determination of the geographic origin of Emmental cheese. Main framework of the project; chemical, biochemical, microbiological, colour and sensory analyses……………………………………………………… 46 6. Analytical methods for the determination of the geographic origin of Emmental cheese. Free fatty acids, triglycerides and fatty acid composition of cheese fat … 54 7. Analytical methods for the determination of the geographic origin of Emmental cheese. Parameters of proteolysis and rheology………………………………….. 69 8. Analytical methods for the determination of the geographic origin of Emmental cheese. Mid- and Near-Infrared spectroscopy……………………………………. 83 9. Analytical methods for the determination of the geographic origin of Emmental cheese. Volatile compounds by GC/MS-FID and electronic nose……………….. 88 10. Analytical methods for the determination of the geographic origin of Emmental cheese. Stable isotope ratios, major, trace and radioactive elements…………….…………………………………………………………….. 93 11. Analytical methods for the determination of the geographic origin of Emmental cheese. Summary of a screening study…………………………………………… 102 I PART III AUTHENTICATION OF EMMENTAL CHEESE: FINAL STUDY AND MODELLING 12. Geographic origin of European Emmental. 1. Characterisation and descriptive statistics…………………………………………………………………………... 112 13. Geographic origin of European Emmental. 2. Pattern recognition and prediction. 128 PART IV APPLICATION TO FURTHER CHEESE TYPES AND CONCLUSIONS 14. Authenticity of Raclette Suisse and Fontina PDO: a feasibility study …………... 136 15. Authenticity of provenance of Swiss cheeses: conclusion of the project, recommendation to food control laboratories and perspective for the future..…… 148 PART V ANNEXES A. Essay for the authentication of Emmental cheese using INDEx coupled to an MS-based electronic nose………………………………………………………… 158 B. Analysis of volatile compounds using a thermodesorption system: first experiences………………………………………………………………………... 160 C. List of publications, poster and oral presentations…………...…...……………… 168 D. List of abbreviations……………………………………………………………… 171 II SUMMARY The current Ph.D. work is divided into four main parts. In the first one, the problematic of volatile compound preconcentration is dealt with to extend the application range of electronic noses (ENs). A review article highlights the advantages of portable equipments such as Solid Phase Micro-Extraction (SPME) and Solid Phase Dynamic Extraction (SPDE). Both showed comparable efficiencies when applied to the SMart Nose equipment, though the second one was less subject to ageing effects. Various canned processed cheese types stored deep frozen were tested for several years as control materials for gas chromatography (long term stability of volatile compounds in cheese-like matrices). These processed cheeses were then used to illustrate the data transferability between two ENs of same type, condition necessary to build usable databases for the daily practice. In the second part of the work, a screening of a great number of analytical methods was carried out to evaluate their potential for the authentication of the geographic origin of Emmental cheese. Twenty Emmental samples from six European regions were investigated on chemical, biochemical, microbiological, physical and sensory parameters. The most promising ones retained for the follow-up of the project were, volatile short-chain acids, chloride, pH-value, total nitrogen, 12%-TCA soluble nitrogen, water soluble nitrogen, copper, sodium, magnesium, zinc, enterococci, obligate heterofermentative Lactobacilli, Lb helveticus, L/D-lactate, succinate, pyruvate, L-leucine-aminopeptidase, δ2H, δ13C, δ15N and δ34S isotope ratios. In the third part, the selected parameters were measured in 110 winter and 73 summer Emmental cheese samples collected in seven European regions, i.e. Switzerland, France Savoie, France Bretagne, France East-Central, South Germany, Austria and Finland. The analytical data was then processed using multivariate statistical analysis leading to pattern recognition and classification according to the geographic origin of the cheeses. Discriminant analysis (DA) and artificial neural networks delivered similar results. DA made it possible a reduction of the number of factors thanks to stepwise backward elimination. In a model including only eleven factors, 95% correct classification in the seven regions was achieved in the Jackknifed validation. Five Swiss Emmental samples out of 70 were misclassified. To improve the classification of the latters, a model with a pairwise approach (Switzerland vs a single foreign region at a time) was proposed. This procedure made it possible to recognise all Swiss samples correctly using fifteen parameters. In the fourth and last part, a targeted screening was applied to two other cheese types, i.e. Raclette and Fontina cheeses. Raclette Suisse® and French Raclette cheese could be easily discriminated using the calcium content and the four stable isotope ratios δ2H, δ13C, δ15N and δ34S. Fontina PDO is a raw milk cheese. Consequently its alkaline phosphatase content was much higher than in the Fontal cheeses, which are industrial imitations manufactured with pasteurised milk. The last chapter is a guide line dedicated to the Swiss
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