complete article - elika

www.heatox.org Project no. 506820

HEATOX

Heat-generated food toxicants: identification, characterisation and risk minimisation

Final report Instrument: Specific Targeted Research Project Thematic Priority: 5. Food Quality and Safety Period covered: From 1 November 2003 to 28 February 2007 Date of preparation: 12 April 2007 Start date of project: 1 November 2003 Duration: 40 months Project coordinator name: Kerstin Skog Project coordinator organisation name: Lunds Universitet

FOOD QUALITY AND SAFETY

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Content 1. Summary ....................................................................................................... 3 1.1 Detailed objectives..........................................................................................5

1.2. HEATOX Partners.................................................................................................................6 1.3. PhD’s ....................................................................................................................................7 1.4. External Panel......................................................................................................................8 2. Formation and Processing.............................................................................. 9

2.1. Chemical mechanisms and kinetics of formation of hazardous compounds.......................9 2.2. New and improved processing technologies to minimise the formation of hazardous compounds.....................................................................................................................................13

2.2.1. Potato....................................................................................................... 13 2.2.2. Bread ....................................................................................................... 19

3. Analysis ....................................................................................................... 21 3.1. Method development.............................................................................................................21 3.2. Analytical results on heat-induced toxicants levels in foods .............................................23 3.3. Analytical method for biomarkers ........................................................................................25

3.3.1. Identification of DNA-adducts in vivo............................................................. 25 3.3.2. Acrylamide metabolites as biomarkers of exposure.......................................... 26

4. Hazard characterisation............................................................................... 30 4.1. In-vivo genotoxicity and carcinogenicity .............................................................................30

4.1.1. Data on the relationship between internal dose and estimated intake of acrylamide in blood samples from a biobank............................................................................ 32

4.2. Tumorigenicity .......................................................................................................................34 4.2.1. Reproductive toxicity .................................................................................. 34 4.2.2. Neurotoxicity ............................................................................................. 35

5. Exposure Assessment .................................................................................. 37 5.1. Monte Carlo Risk Assessment...............................................................................................37

5.1.1. Dietary exposure assessment....................................................................... 37 5.1.2. Scenario-analysis of effect of new processing techniques.................................. 37 5.1.3. Brand loyalty ............................................................................................. 38 5.1.4. Home cooking............................................................................................ 38 5.1.5. Intake in British and Dutch infants ................................................................ 39

5.2. Training on probabilistic exposure assessment ..................................................................39 5.3. Estimation of breast cancer risk from human exposure to heat-induced toxicants.........39 5.4. Measurement of internal doses of acrylamide and glycidamide – a tool for exposure assessment and risk assessment.................................................................................................41

5.4.1. Haemoglobin adducts as a measure of internal dose........................................ 41 5.4.2. Bioavailability of acrylamide in food .............................................................. 41 5.4.3. Endogenous formation of acrylamide............................................................. 41 5.4.4. Inter- and intra-variation of internal doses in humans...................................... 41 5.4.5. Relationship between internal dose and estimated acrylamide intake from food ... 42

6. Risk assessment and risk communication .................................................... 43 6.1. Risk assessment ....................................................................................................................43

6.1.1. Models for extrapolation of cancer risk........................................................... 43 6.1.2. Final Risk Characterisation........................................................................... 43

6.2. Risk communication ..............................................................................................................45 6.2.1. Risk Communication Strategy....................................................................... 46

Acknowledgement………………………………………………………………………………….49 Publishable results of the Final plan for using and disseminating the knowledge……..………… 50 Dissemination of Knowledge……………...…………………………………………………........54

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Final report – Contract FOOD-CT-2003-506820

Heat-generated food toxicants: Identification, Characterisation and Risk Minimisation (HEATOX)

1. Summary

Heating food gives many advantages – it adds taste, colour and texture and minimises harmful germs. Flavour and aroma compounds are produced via the Maillard reaction but at the same time various health hazardous compounds may form. The main route for acrylamide formation goes via the Maillard reaction between reducing sugars (glucose and fructose) but also sucrose, and the amino acid asparagine. The HEATOX project involved 24 partners in 14 countries. The focus of the HEATOX project was health risks associated with hazardous compounds, for example acrylamide, in heat-treated carbohydrate-rich foods. The main objectives were to estimate health risks that may be associated with hazardous compounds in heat-treated food and to find cooking/processing methods which minimise the amounts of these compounds, thereby providing safe, nutritious and high-quality foodstuffs. Among the important results are recommendations to consumers, restaurants and the food industry on how to minimise the amounts of heat-generated toxicants in foods, while ensuring product quality from a nutritional and sensory point of view.

The HEATOX project started on the 1st November 2003 and had a duration of 40 months. Project web-site: www.heatox.org Co-ordinator: Kerstin Skog, Lund University, Sweden Department of Food Technology, Engineering and Nutrition e-mail: [email protected] Scientific Officer: Jürgen Lucas, e-mail: [email protected] The HEATOX project has dealt with questions regarding heat-generated food toxicants e.g.

• In which foods are these compounds mainly found? • How are they formed? • Do they constitute a health risk? • How can we measure/control the amounts produced? • How much is consumed? • What are the effects on the human body? • How can they be avoided? • Is there a cooking method to be recommended?

This European Research Project is supported within the European Commission's 6th Framework Programme on Research, Priority 5 on Food Quality and Safety (Contract n° Food-CT-2003-506820 Specific Targeted Project). This publication reflects the author’s views and not necessarily those of the EC. The information in this document is provided as is and no guarantee or warranty is given that the information is fit for any particular purpose.

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Modern science has showed that heating of meat and other protein rich foods can generate various kinds of potentially hazardous compounds, some of which are genotoxic and carcinogenic. The focus of the HEATOX project was health risks associated with hazardous compounds in heat treated carbohydrate-rich foods where substantial amounts of acrylamide and similar compounds can be formed. HEATOX explored their mechanisms of formation, impact of raw material composition, inhibiting factors, cooking and processing methods in industry and households with the to aim control and minimise the formation of hazardous compounds. Acrylamide was given particular emphasis in the HEATOX project. Other compounds representing potential health hazards, such furans, are also formed during heating, thus it is important that efforts in reducing acrylamide exposure at the same time do not increase the formation of other hazardous compounds. Validated methods for food analysis and exposure biomarkers have been developed. Different hazards have been explored and characterised in various toxicological models, for example genotoxicity, carcinogenicity, neuro-developmental and reproductive toxicity. Molecular characterisation by toxicogenomics has been performed. Experimental animal and cell systems and also humans were studied. Particularly the relevance of low dose exposure, bioavailability and extrapolation from experimental systems to humans was addressed using biomarkers of exposure and effect. The exposure assessment and data on hazard characterisation, including data generated outside HEATOX, were combined in a risk characterisation of intake of heat-treated carbohydrate rich foods. To support and strengthen the project profile, a risk communication strategy was developed early in the project. A tailored action plan was implemented successively during the whole project period. The ultimate aim was to ensure that project deliverables were as useful and focused as possible and that the dialogue within HEATOX and between HEATOX and the outside world was effective. The HEATOX project co-operated with other research activities carried out by other groups around the world, e.g. the project was linked with the COST 927 action. Dissemination of the project results was a key issue, which involved for example publications, conferences, and the HEATOX workshops.

Formation Exposure Assessment Hazard characterisation Analysis

Risk Assessment

Management, Communication, Dissemination and Training

Formation

Industrial Processing

Household Cooking

External Exposure

Internal Exposure

DNA damage

Non- genetic damage

Mutation, Cancer

Fertility

Neurological effects

Other Health

Effects

Project overview

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1.1 Detailed objectives: I. To elucidate the chemical reaction mechanisms involved and to develop and validate new/improved production methods and technological minimisation strategies ♦ Effects of variation in the composition of the raw material ♦ Reaction pathways and mechanisms, inhibiting/enhancing compounds, and kinetics ♦ Cooking and processing methods for the food industry, catering facilities and households Expected results: Recommendations on how to limit the formation of heat-induced toxicants in order to minimise the formation of heat-induced toxicants, while ensuring product quality from safety, nutritional and sensory points of view. II. To provide validated analytical methods (i.e. screening and confirmatory) and quality-controlled analytical data needed for the execution of the other tasks • Quantitative analytical methods for biomarkers and for toxicants in foods • Identification methods and data on new toxicants or known toxicants in new food matrices • Quality-controlled data on toxicant levels in foods Expected results: Validated analytical methods for a wide range of heat-induced toxicants and for biomarkers of exposure. III. To perform hazard characterisation ♦ Neurodevelopmental toxicity, mechanisms of tumourgenicity, and mechanisms of reproductive

toxicity ♦ Bioavailability in animals and humans determined by different biomarkers for internal dose and

effect ♦ Dose-response relationships in the low-dose region Expected results: Characterisation of toxic effects including their mechanisms and dose-response relationships. IV. To assess the exposure of heat-induced toxicants ♦ Improved estimation of the intake of heat-induced toxicants based on analysis of cooked foods and

food intake data, especially concerning variability and risk groups ♦ Comparison of dietary intake data regarding heat-induced toxicants with data on the internal dose

(heat-induced toxicants adduct levels, e.g. DNA-adduct levels) ♦ Elucidation of the role of possible endogenous formation of heat-induced toxicants ♦ Assessment of the possibility of using statistics for conclusive cancer epidemiology studies Expected results: Validated data on heat-induced toxicants exposure and intra- and inter-individual variation in exposure. V. To perform risk assessment and communicate the results of the project ♦ Assessment of the health risk associated with the current exposure to known heat-induced

toxicants (risk characterisation) and possibly new compounds, also taking into account existing information on hazards from sources outside the project

♦ Comparative risk/benefit assessments of heat-treated foods by linking results from animal and human studies to human health risk

♦ Targeted recommendations and guidelines to all important user groups ♦ Broad information to the general public, the food industry and its different relevant

associations/federations, national food agencies, consumer groups/associations, EFSA, different Commission services (e.g. DG Research, DG Health and Consumer Protection, DG Agriculture, DG Enterprise, etc.) on the project, its results and their implications including also future aspects.

Expected results: Increased knowledge and harmonised agreement on the possible risks of heat-induced food toxicants and recommendations on how the toxicant levels can be controlled.

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1.2 HEATOX Partners

P 1 Lund University (Sweden) • Kerstin Skog, Co-ordinator P 2 Graz University of Technology (Austria) • Michael Murkovic P 3 The University of Reading (UK) • Don Mottram P 4 Swedish University of Agricultural Sciences (Sweden) • Per Åman P 5 University of Bologna (Italy) • Marco Dalla Rosa P 6 Swedish Institute for Food and Biotechnology (Sweden) • Hans Lingnert P 7 Wageningen University (The Netherlands) • Tiny van Boekel P 8 Central Science Laboratory, York (UK) • Laurence Castle P 9 Swedish National Food Administration (Sweden) • Karl-Erik Hellenäs P10 Institute of Chemical Technology Prague (Czech Republic) • Jana Hajslova P11 Agrotechnology and Food Innovations (The Netherlands) • Charon Zondervan P12 University of Barcelona (Spain) • Maria Teresa Galceran P13 TÜBÝTAK-Marmara Research Center (Turkey) • Hülya Ölmez P14 Stockholm University (Sweden) • Margareta Törnqvist P15 National Food Institute (Denmark) • Henrik Frandsen P16 Norwegian Institute of Public Health (Norway) • Jan Alexander P17 RIKILT Institute of Food Safety (The Netherlands) • Jacob van Klaveren P18 German Institute for Human Nutrition (Germany) • Hansruedi Glatt P19 University of Leeds (UK) • Bronek Wedzicha P20 BEUC, European Consumers' Organisation (Belgium) • Barbara Gallani P21 National Veterinary Institute (Norway) • Helga Odden Reksnes P22 University of Zürich (Switzerland) • Hanspeter Naegeli P23 University of Chile (Chile) • Lilia Masson P24 Queens’ University Belfast (UK) • Chris Elliott

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1.3 PhD’s Exchange of young scientist has been encouraged within the HEATOX project; several young researchers have visited other HEATOX partners for 1-4 weeks to learn new methods, plan collaborative work, discuss results etc. The successful integration of the young researchers with senior researchers and cooperation between the different groups has contributed to interdisciplinary relations and building of networks. Several partners participate in Cost Action 927 “Thermally Processed Foods: Possible Health Implications”, co-ordinated by V Fogliano (University of Naples, Italy), which has provided additional opportunities for student exchange. Special emphasis has been put on communication training for PhDs and young researchers. At the semi annual meetings, seminars have been arranged on the topic of risk communication and dissemination in relation to their HEATOX work, for example short oral presentations, posters and also participation in the HEATOX workshop. PhD students/Young scientists working in the HEATOX project: German Institute of Human Nutrition (Germany) • Yasmine Sommer Graz University of Technology (Austria) • Kristina Bagdonaite Institute of Chemical Technology Prague (Czech Republic) • Lenka Dunovska Leeds University • Jonas Mojica Lazaro Lund University (Sweden) • Peter Viberg, Gunilla Viklund National Food Institute (Denmark) • Pelle T Olesen Norwegian Institute of Public Health (Norway) • Thomas Bjellaas, Siri Helland Hansen, Hege Benedikte Ölstörn RIKILT Institute of Food Safety (The Netherlands) • Anika de Mul Stockholm University (Sweden) • Anna Vikström Swedish National Food Administration (Sweden) • Louise Durling, Erik Petersson Swedish University of Agricultural Sciences (Sweden) • Arwa Mustafa The University of Reading (UK) • Mei Yin Low University of Barcelona (Spain) • Soubhi Altaki, Erika Teixido University of Bologna (Spain) • Alessandro Angioloni, Mara Bacchiocca, Andrea Gasparri, Pietro Rocculi University of Zürich (Switzerland) • Flurina Clement, Ramiro Dip Wageningen University (The Netherlands) • Jeroen Knol Queen’s University Belfast (UK) • Andrew Preston

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1.4 External Panel The External Panel constituted the core risk communication network and provided valuable discussion with HEATOX’ stakeholders. In charge: Karl-Erik Hellenäs. Members: Consumer • Barbara Gallani (from June 2005)/Beate Kettlitz (until March 2005), BEUC, BE

Industry • Beate Kettlitz (from March 2005)/Domenique Teaymans (until March 2005), CIAA, BE • Sam Lalljie, ILSI, BE • Julia Gelbert, BLL/ZUTECH, GE

Research • David Lineback, JIFSAN, USA • Erland Bråthen (until 2005), MATFORSK, NO • Vincenzo Fogliano, COST-927, IT • Elke Anklam (until July 2006), JRC irmm, BE

Authorities • Almut Bitterhof (from spring 2006)/Martin Slayne (until spring 2006), DG Sanco, BE • Claudia Heppner, EFSA, Contam panel, IT • Lutz Dehne, BfR, DE • Wendy Matthews (from 2004)/Karen Goonan (until 2004), FSA, UK • Sara Henry, FDA, USA • Lauren Jackson, FDA, USA Management & Information Flow

Management Formation, Processing

Analysis Hazard characterisation

Exposure Risk assessment, communication

Scientific Committee

Coordinator

EC- DG Research

External consulting panel

(One representative per partner incl. Coordinator)

(Theme leaders and coordinator)

Technical Project Assistant

WP 1 WP 2.1, 2.2 WP 3.1, 3.2, 3.3

WP 4.1, 4.2, 4.3

WP 5.1, 5.2, 5.3

WP 6.1, 6.2

Board of Managers

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2. Formation and Processing Wageningen University (The Netherlands), University of Leeds (United Kingdom),

The University of Reading (United Kingdom), Graz University of Technology (Austria), Lund University (Sweden), Agrotechnology & Food Innovations B.V. (The Netherlands), University of Chile, University of Bologna (Italy),

Swedish Institute for Food and Biotechnology (Sweden), Swedish University of Agricultural Sciences (Sweden)

2.1. Chemical mechanisms and kinetics of formation of hazardous compounds The Maillard reaction between amino acids and sugars is essential for the development of desirable colour and flavour in baked and fried foods. However, the Maillard reaction has been shown to generate acrylamide at the same time, through the reaction of the amino acid asparagine with reducing sugars. Thus, acrylamide is formed by the thermal reaction of natural food components, when moisture levels become low. A proposed mechanism for acrylamide formation (see Figure 1) involves the formation of a Schiff base from the reaction of a carbonyl compound with asparagine. Decarboxylation of the Schiff base, in a Strecker-type reaction, gives an unstable intermediate that can hydrolyse to 3-aminopropamide, which on elimination of ammonia yields acrylamide. Alternatively the decarboxylated Schiff base could form acrylamide via elimination of an imine.

Figure 1. Proposed mechanism for the formation of acrylamide in heat-treated foods.

O NH2

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NH2

O O

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ASPARAGINE REACTIVE CARBONYL FROM

MAILLARD REACTIONSCHIFF BASE

STRECKERALDEHYDE3-AMINOPROPANAMIDE ACRYLAMIDE

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The formation of acrylamide in food is under kinetic control; this means that the amount which is formed depends on the rate of reaction and the time for which it has been allowed to proceed. Reaction kinetics relates to the rate limiting steps in chemical reactions; these are the control points. For this reason, an understanding of the factors which control the rate limiting steps in acrylamide formation offers opportunity to control the extent of its production. Chemical mechanisms and kinetics of formation were studied, based on reactions between sugars and amino acids via the Maillard reaction. Equimolar solutions of glucose and asparagine (0.1 and 0.2 M) were prepared in phosphate buffer (0.1 M) with different pH (4.5, 5.5, 6.0 and 6.8). Samples were heated at 120-200°C for 0-128 min. A mechanistic kinetic model has been derived for the aqueous model systems from the proposed reaction network (Figure 2). For each reaction step a differential equation was set up, and translated into a mathematical model.

Figure 2. Proposed reaction network for the formation of acrylamide from asparagine and glucose/ fructose through early Maillard reaction. Product X: product(s) formed from acrylamide breakdown.

Multiresponse modelling using non-linear regression with the determinant criterion was used to estimate model parameters, i.e. the rate constants and activation energies based on the experimental data obtained from the experiments with the aqueous model systems (Table 1).

Table 1. Estimates of rate constants (k) at 120, 140, 160, 180 and 200 ºC and activation energies (Ea) ± 95% Highest Posterior Density (HPD) interval as found by kinetic modelling for the proposed kinetic model for pH 6.8.

k (10-3 min-1 or·10-3 mol·min-1) 120 ºC 140 ºC 160 ºC 180 ºC 200 ºC Ea (kJ/mol)

k1 0.131 ± 0.025 0.308 ± 0.049 0.668 ± 0.13 1.35 ± 0.36 2.58 ± 0.89 57.6 ± 8.0

k2 4.98 ± 1.2 16.7 ± 3.0 50.1 ± 9.6 136 ± 35 341 ± 114 81.7 ± 8.6

k3 0.0819 ± 0.045 0.369 ± 0.14 1.45 ± 0.44 5.04 ± 1.6 15.8 ± 6.4 102 ± 14

k4 0.176 ± 0.081 0.712 ± 0.23 2.53 ± 0.51 8.05 ± 1.2 23.2 ± 4.3 94.4 ± 11

k5 15.7 ± 3.5 28.4 ± 4.6 48.7 ± 5.9 79.6 ± 8.9 125 ± 16 40.1 ± 5.0

k6 7.96 ± 5.1 28.1 ± 13 88.1 ± 25 250 ± 45 650 ± 136 85.1 ± 14

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Figure 3 shows how the model fits the experimental data for the formation of acrylamide in the aqueous reaction system at pH 6.8. The proposed model gives a reasonable estimation for the formation of acrylamide in an aqueous model system. The mechanistic kinetic model derived in this study is an important step into the realization of a tool that can be used to predict how acrylamide reduction in foods containing asparagine and reducing sugars can be accomplished.

Figure 3. Model fit (lines) to experimental data (symbols) of acrylamide concentration for glucose/ asparagine (0.2 mol/l, pH 6.8) aqueous reaction system heated at 120 (blue), 140 (red), 160 (purple), 180 (green) and 200 °C (light blue).

Another kinetic model for the formation of acrylamide in foods (potato, wheat and rye products) involves steps which lead to the destruction, binding or physical ‘loss’ of acrylamide. Our ability to formulate correctly the kinetic model for acrylamide levels in foods thus depends critically on us being able to formulate the kinetics of these subsequent processes. This is particularly important with regard to the control (i.e., reduction) of acrylamide levels because such ‘losses’ could be maximised through adjustment of the reaction conditions. Furthermore, there are views that perhaps much more acrylamide is actually formed in foods than is measured, and an understanding of the fate of acrylamide is important so that we might be able to identify the chemical nature of any ‘bound’ acrylamide and the stability of any reaction products which are formed. If acrylamide is being formed in much greater yield than so far observed, there is also a danger that a change in the reaction conditions could alter the balance between the rates of formation and of loss, so that much higher yields are obtained. We believe, therefore, that consideration of the kinetics of acrylamide loss is fundamental to the derivation of the kinetic model. The kinetics of the reaction of acrylamide with glycine and proline were determined in aqueous systems as a function of pH and temperature, and the relative reactivities of all 20 amino acids measured at pH 5.5. Maltodextrin gels were prepared containing acrylamide and the amino acids, and were freeze-dried before being heated to 180 °C. The experiments in aqueous systems have shown the relevance of Michael addition type reactions of amino acids with acrylamide. Acrylamide reacted more readily with glycine than with Maillard precursors in solution and this reaction presented second order kinetics, i.e.,

]lyk[acryl][gdt

d[acryl]=−

However, glycine was not the most reactive amino acid towards acrylamide; proline and cysteine showed much higher rates of reaction in comparison with other amino acids. pH was a significant factor in the reaction; glycine and proline react by a common mechanism in which there exists an acid-base catalytic effect by a species with pKa=5.2-5.5.

])glyNH[(][A 21-

0−+= kkk

where k is the apparent first order rate constant and A- is the suggested catalyst.

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The maltodextrin dry solid system demonstrated reactivity of amino acids towards acrylamide when heated in the absence of water at 180 ºC. The rate of loss of acrylamide in this system was very similar to that in starchy foods. In this model, most of the amino acids showed similar reactivity towards acrylamide, with the exception of lysine, which was higher. Lysine is also one of the most reactive amino acids in the Maillard reaction. The effect of temperature on the amino acid-acrylamide reaction followed the Arrhenius relationship, showing the highest Ea (83 kJ mol-1) for the reaction of acrylamide alone in the matrix. The reaction of glycine and proline with acrylamide showed lower but similar Ea of approximately 70 kJ mol-1. Our data indicates that acrylamide is lost from food by reaction with amino acids. This augments the already established effect of amino acids (e.g., glycine) on acrylamide formation by competing with asparagine for a key precursor of acrylamide, and explains published reports describing the effects of amino-containing food ingredients on acrylamide residues. Maltodextrin proved to be a good model system in which to study reactions of acrylamide with amino acids in the dry state, as evidenced by rates of reaction of acrylamide which are similar to those observed in foods. The relationships between the formation (and loss) of flavour compounds and acrylamide were measured in potato and rye. Simple cakes were prepared from flour and water and were heated at 180 C, from 10 to 60 min. The flavour compounds studied were Strecker aldehydes, which are formed relatively early in the Maillard reaction and may react further, and alkylpyrazines, which may be considered as end-products. The data obtained was used to create a kinetic model, which was then used to predict the effect of glycine on flavour and acrylamide formation in potato cakes. The relationships between the formation (and loss) of flavour compounds and acrylamide were different in potato and rye. In both potato and rye, Strecker aldehydes formed and decomposed in a similar way. However, in rye the rate of formation of the Strecker aldehydes was similar to that for acrylamide, whereas, in potato, acrylamide formed at a slower rate than in rye. Pyrazine formation in rye and potato mirrored acrylamide formation. However, acrylamide levels decreased on prolonged heating whilst pyrazine levels were maintained in both rye and potato. It appeared that moisture levels below 5% were needed for pyrazines and acrylamide to form in both foods. Potato cakes contained higher initial moisture levels and hence formation of both acrylamide and pyrazines occurred later in the cooking process for potato than for rye. The Maillard reaction could be responsible for potential toxicants other than acrylamide. Two databases, each containing chemical and toxicity data, have been compiled in Microsoft Access for Maillard compounds and for lipid-derived compounds. The Maillard database lists approximately 570 volatile compounds, while approximately 200 volatile compounds are listed as products of heated lipid systems. A molecular modelling approach has been used to assess the potential toxicity of each compound. Both databases are fully searchable and contain the following information: • Chemical name, Chemical structure, CAS number • Other names • Foods in which the compound is present • Foods in which the compound is present at levels above 1 ppm • Reaction mixtures in which the compound is present • EU Approved Flavourings Register FL No. • Predicted carcinogenicity probability • Predicted mutagenicity probability • Predicted rat oral LD50 (mg/kg) • Predicted chronic lowest observed adverse effect level (LOAEL) (mg/kg) • Predicted skin sensitisation probability The databases provide a useful source of information for chemists and toxicologists. They are an important starting point for future research into toxic compounds formed during cooking. The data bases can be found at www.heatox.org, Achievements.

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The formation of acrylamide during the roasting process of coffee was investigated in detail in lab scale using a roasting device that employs convection roasting. The model system was a mixture of asparagine with glucose or sucrose using a heating device that allowed the constant heating of small samples (max 300 mg) up to 300 °C with an efficient heat transfer. Studies using 13C-labelled asparagine showed that this amino acid is directly converted to acrylamide. Intermediates were synthesised (N-glucosyl-asparagine and the asparagine Amadori compound) that were converted to acrylamide by heating. At high temperatures – which are used during roasting of coffee – these two compounds could be identified as intermediate reaction products. A statistical analysis suggests that asparagine is the limiting compound in reactions leading to acrylamide in coffee. Sucrose is present in green coffee in concentrations up to 9%. A statistical analysis suggests that asparagine is the limiting compound in reactions leading to acrylamide in coffee. In coffee shorter roasting times and lower temperatures are leading to higher acrylamide concentrations. This means that darker roasted coffee – higher degree of roasting – has lower acrylamide concentrations. HMF (hydroxymethylfurfural) is formed in coffee very quickly which means that within 2 min a maximum concentration is achieved. After this time a quick degradation occurs to levels significantly lower (ca 100 µg/g). This means that HMF is not heat stable and reacts further during the roasting process to substances that are not known now. HMF is formed from sucrose with and without the participation of amino acids.

2.2. New and improved processing technologies to minimise the formation of hazardous compounds 2.2.1. Potato Potato has been a staple food for many decades and mainly consumed as boiled or fried potatoes. Our dietary habits have changed and today we are used to many different potato dishes and there are a number of potato products on the market. Potatoes are rich in starch, and the aim of heating potato is to make them palatable and to gelatinise the starch to make it available for digestion. At the beginning of the project a mindmap was made, se Figure 4 below. Such a mindmap serves to find relationships between the numerous variables that – in this case – potentially influence various quality parameters of French fries. With a special focus on acrylamide, the mindmap gave clusters of variables that were somehow related. These clusters of variables formed the basis of our three-year research programme.

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2.2.1.1. Heating model: Fry Simulator – French fries A computational model was made to estimate the impact of various processing parameters on the formation of acrylamide in French fries. The model uses a cross section of a French fry of infinite length and is made up from a 40*40 grid. The model was validated by measuring temperature and water profiles in time and at various positions inside the French fries. At a later stage also acrylamide measurement were performed on parts of the fries. The latter results showed that although the trends are correctly predicted in the Fry Simulator, its value in absolute sense is limited. Moreover, since the model only calculates acrylamide in the middle, it leaves out the acrylamide that is formed on both ends of the fries. From experimental data it is well known that the highest levels are found at both ends. 2.2.1.2. French fries Experiments were performed with potatoes grown in the Netherlands. Batches of three varieties of 1000 kg each (Asterix, Bintje, Lady Olympia) were supplied by a potato processor, shortly after the harvest and were stored at 8°C until the end of June. In the varieties Asterix and Bintje the sugar concentrations increased during a 6 month storage period. The sugar contents of the Lady Olympia potatoes decreased during the storage. We also found that the acrylamide levels were positively correlated to the levels of reducing sugars. We found that acidic blanching prior to industrial frying is an efficient way to reduce acrylamide during final product preparation at home or in restaurants and caterers. When using 0.5% of citric or lactic acid, acrylamide levels are reduced some 80% while expert panels do not detect any acidic aftertaste. Other methods of introducing small amounts of acid onto the surface have been considered, such as spraying acid solutions on frozen fries. Such methods were expected to be no more efficient than acidic blanching but much more difficult to implement. 2.2.1.3. Potato crisps Panda Potatoes variety, crops 2004, 2005, 2006 were used for experiments and were pre-treated before frying at 120, 150 and 180 ºC: washing with distilled water, blanching at 90 ºC per 1 minute and immersion in dilute citric acid solution 1-0.25% for 1 hour. Experiments were performed in vacuum and at atmospheric pressure. Vacuum frying with and without pre-treatments was shown to produce low level acrylamide crisps, see Table 2. It is possible to improve their quality characteristics. The pre-treatments represent a practical industrial alternative to reduce acrylamide level in potatoes crisps.

Table 2: Effect of pre-treatments, frying methods and frying temperatures on acrylamide content and acceptability score (1-7) of crisps.

Pre-treatment Frying method

Blanching

Immersion citric acid

0,25% Atmospheric Vacuum

(50 mm Hg) Acrylamide

Acceptability

90ºC/1 min 1h Temp.ºC Temp.ºC ppb Score*

+ + 150 - 6 4,3 + + 160 - 5 4,3 + + 170 - 30 4,4 - - 150 - 87 2,1 - - 160 - 233 2,9 - - 170 - 760 2,9 + + - 150 ND 3,3 + + - 160 ND 3,5 + + - 170 4 3,5 - - - 150 80 4,6 - - - 160 218 5,3 - - - 170 267 5,1

*Score 1-7, with 7 as the best ND = Not Detected

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Blanching and change of pH by immersion of sliced raw potatoes in citric acid solution (0.25%), had a strong influence for reducing the acrylamide content in crisps fried at atmospheric pressure and in vacuum. Acrylamide content of potatoes crisps with pre-treatments fried in vacuum were very low between non detected levels up to 4 ppb. At atmospheric pressure with pre-treatments, the acrylamide content was between 6 up to 30 ppb. Without pretreatments, the acrylamide content of the crisps varied between 80 – 267 ppb for vacuum frying and at between 87 – 760 ppb for frying at atmospheric pressure. The sensory evaluation (trained and consumer panel) obtained as acceptability score ( 1 – 7), was better for the crisps with pretreatments fried at atmospheric pressure than in vacuum, 4.3 compared with 3.5. A commercial crisp sample sensory evaluated at the same time scored 4.8 with an acrylamide content of 536 ppb. The sensory panel concluded that the best crisps were obtained in the samples corresponding to control assays, submitted to vacuum frying processing at 160ºC and 170 C. Considering the assays with pre-treatments, the vacuum fried crisps got a better evaluation than the crisps fried at atmospheric pressure. Conclusion: Vacuum frying is a processing method not commonly used in the industry, however, it can be a very good alternative in the near future (acrylamide free crisps)

Raw Potatoes, Var Panda Blanching Citric Acid Immersion

Slices potatoes

Vacuum Fryer

Potato Crisps

Photos: Lilia Masson

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2.2.1.4. Raw materials selection

Potato field in the south of Sweden. Photo: K. Olsson, Svalöf Weibull

Well characterised Swedish-grown potatoes, including two breeding clones that are resistant to low-temperature sweetening were studied and batches were obtained from three consecutive years. The content of free amino acids and sugars in potato were studied in potatoes from three growing seasons. In some potatoes, the levels of reducing sugars were practically the same the first two years, but almost doubled the third year, while in another variety; the level of reducing sugars decreased to half the second year and increased the third year. The concentrations of asparagine were generally not so much affected by the growing season. These observations make it difficult to draw conclusions on which variety processing conditions that are best for crisp production because this may vary with year. Frying conditions were discussed with crisp producers and producers of frying equipment to establish laboratory conditions that would resemble industrial conditions. For industrial crisp production, potato slices are usually passed on a belt through a continuous fryer with a starting temperature around 180-190 C and an end temperature of around 150 – 175°C. Quality parameters such as crispiness and colour have to be taken in account. A water content below 2% is considered to give a good crispiness and shelf-life. Using genotypes with a wide range of precursor concentrations, we found that the asparagine levels affect the formation of acrylamide to a higher degree than earlier studies have shown. Crisps made from potatoes with the lowest asparagine content, generally had the lowest concentration of acrylamide regardless of storage temperature. In agreement with work by others, we found that storage at 4°C enhanced the concentration of sugars in he tubers and thus the acrylamide content compared with storage at 8°C. Blanching before frying was shown to efficiently reduce the acrylamide content in crisps. For potatoes stored at 4 C, the acrylamide content was generally reduced by at least 50%. The mass transport of precursors during heating may be important for acrylamide formation, but this requires further investigation.

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2.2.1.5. Home cooking Much work has been devoted to the mitigation of acrylamide in potato crisps and French fries, but literature data on acrylamide in home-cooked potato dishes is scare. Several home-cooking experiments were performed. Roasted potato wedges and pan fried boiled potato dices (a common Swedish dish) had an appetising appearance; the wedges were soft with a golden yellow surface. Acrylamide was found in all cooked samples. Blanching before oven roasting made it possible to reduce the heating time in the oven and yet obtain potato wedges with similar eating quality. Blanching combined with a shorter roasting time is an easy and efficient way of reducing the acrylamide content in oven roasted potato wedges.

The effect of frying in a commercial available domestic fryer was studied in relation to acrylamide formation in French fries. It was found that the frying temperature drops drastically and slowly reached its original temperature. The drop in temperature depends of the product/oil ratio and makes it difficult to define an initial oil temperature as a measure to reduce acrylamide.

The results showed that higher initial oil temperature causes a faster recover of oil temperature during frying, as well as a greater acrylamide formation rate. The choice of a lower initial frying temperature lowered the acrylamide content of the product. At the optimal cooking time in terms of culinary quality, the lower the potato quantity the lower acrylamide content was found. Our results underline that the kind of frying equipment (heating power), frying time, oil temperature, amount of potato immersed in the hot oil (product/oil ratio) have to be taken into account for optimisation of product quality (colour, texture, water and oil content) and acrylamide content in French fries. Since par-fried French fries do not contain acrylamide when leaving the factory, it is important to instruct users (both consumers and the out-of-home channel) to cook the products in a proper way. Time and temperature instructions should be clearly stated on the package, preferably at somewhat lower temperatures. Instructions should also consider the volume of product to be cooked. For frying the ratio of oil-to-product should be clear. For oven cooking, the electrical power-to-product ratio should be indicated. Secondly, we found that power of household equipment (ovens and deep fryers) varies and should be taken into account as well. Producers of consumer appliances could improve their product by using better time and temperature controls. Most important perhaps is that many consumers overcook French fries, thereby increasing acrylamide levels substantially, simply because of specific liking (darker and dryer end product) or routine behaviour or sometimes negligence.

Domestic fryer. (Photo: University of Bologna)

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2.2.2. Bread Studies have shown that asparagine is the limiting factor for formation of acrylamide in bread, and that asparagine is mainly concentrated in cereal bran. This means that high fibre flour will result in higher acrylamide content than sifted flour. New fermentation techniques could help in break down of asparagine and thus lowering acrylamide. The effects of asparagine and fructose, added to the dough, were studied in an experiment with a full factorial design. More than 99% of the acrylamide was found in the crust. Added asparagine dramatically increased the content of acrylamide in crusts dry matter (from about 80 µg/Kg to between 600 and 6000 µg/Kg) while added fructose did not influence this content. Both time and temperature increased the acrylamide content in crust dry matter, and a significant interaction was found between these two factors. When baked at different conditions with the same ingredients, a highly significant relationship between colour and acrylamide content was found. Added asparagine increased acrylamide content in yeast-leavened breads while added glycine decreased the content. The more asparagine in the dough, the stronger was the reducing effect of glycine. When glycine was applied on the surface of the fermented dough, also a significant reduction of acrylamide content in the bread was found. Addition of glycine but not of asparagine caused an increased browning reaction during baking. Fermentation of doughs made with different milling fractions showed that most of the asparagine was used up after 2 hr of fermentation with bakers yeast. Sourdough, on the other hand, did not reduce the content of free asparagine as efficiently but had a strong negative impact on asparagine utilization by the yeast. This indicates that this type of fermentation may result in bread with higher acrylamide content than breads fermented with yeast only. Short fermentation time compared to longer fermentation reduced acrylamide content in bread made with whole grain wheat by 87%. For bread made with rye bran, the corresponding reduction was 77%. Baking time and temperature increased acrylamide content in rye crisp bread. A significant interaction between time and temperature of baking was found. Added asparagine had a significant effect on the formation of acrylamide but fructose had not. There was a correlation between acrylamide content and colour of the milled bread in the time-temperature experiment but this correlation was not observed in the experiment with added precursors. Added oat-bran concentrate with high content of mixed-linkage β-glucan did not influence the acrylamide content in the breads. 2.2.2.1. New baking techniques New baking techniques such as air jet impingement and infrared radiation baking has focussed on wheat based bread. Various baking technologies have been studied: Traditional baking; Steam baking; Jet air impingement baking, and Infrared radiation (IR) baking.

Baking by infrared radiation. (Photo: Swedish Institute for Food and Biotechnology)

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First the influence of baking parameters on acrylamide formation and bread characteristics at conventional baking was evaluated. We could show that the highest acrylamide levels normally were found in the outer part of the crust and that the levels increased with increased baking temperature (200-260 C) and prolonged baking time (10-25 min). However, at the highest temperature-time combination (260°C; 20 min) the acrylamide level decreased and was lower in the outer crust than in the inner crust. From this knowledge base, we worked along two lines: the application of steam to traditional baking and alternative heating techniques, such as infrared radiation and air jet impingement. Steam baking We studied the effects of using steam during the final part of baking. The reference baking parameters were 200 C baking temperature for 20 minutes, and this was compared to baking when steam was introduced after 5, 10, or 15 minutes of baking. The steam also lowered the temperature of the bread, so for comparison baking experiments were performed where similar temperature profiles were obtained by temperature adjustments in the oven. Both the steam baking and the “falling temperature” baking substantially lowered the acrylamide levels in the final bread crust. However the crust colour was more influenced in the falling temperature breads than in the steam baked breads. With steam baking it was possible to find conditions that gave bread of the same colour level as that achieved by traditional baking, but with considerably lower acrylamide levels. With falling temperature baking, however, though acrylamide levels were lowered, the bread colour was lighter. Using sensory analysis, we could show that it was possible to bake bread (using steam the final 5 minutes) that was not significantly different from the control regarding odour, appearance, texture, and flavour, but having 40% lower acrylamide level. Alternative baking techniques The effect of new baking techniques, such as air jet impingement and infrared radiation baking, on bread crust formation and characteristics was initially studied. From the knowledge of such experiments baking conditions were defined to reach similar crust colour with the various baking techniques. It was obvious that the acrylamide content in the bread crust could be reduced by using alternative baking technologies. Reduced levels were obtained with infrared radiation baking as well as with air jet impingement baking. Preliminary experiments were also performed to study the flavour formation in bread baked by these new technologies. It turned out to be a close relationship between a large number of volatiles and the formation of acrylamide for the baking methods studied, especially those volatile and odorous compounds that are known to be formed via browning reactions in the bread crust, e.g. Strecker aldehydes and alkyl pyrazines. Obviously, a baking method that resulted in low acrylamide content in these experiments also tended to result in low contents of important flavour substances. We then further focussed on IR baking to evaluate how the IR baking can be optimized with regard to acrylamide minimisation and sensory product quality. Two different IR wavelength regions (one in the shorter range, IRS; and one in the medium range, IRM) and two effect levels (100% and 50%) were compared, using traditional baking as a reference. Flat bread cakes were used for the study. We followed the development of crust colour, acrylamide, flavour compounds and sensory characteristics with baking time at the various baking conditions. One important finding from these experiments was that with IR baking it is possible to obtain a sensory profile almost identical to that of the conventionally baked bread, but with considerably lower acrylamide content. A reduction of the acrylamide content by 60% could be shown. The main conclusion is that it is possible to reduce the acrylamide content while retaining the sensory quality both by introducing steam in traditional baking and by using new alternative baking techniques. In this work it was possible to reduce the acrylamide content by 40% in white bread, by applying steam during the final five minutes of baking, with no significant changes of the sensory quality. Using infrared radiation heating it was possible to reduce the acrylamide level in flat bread cakes by 60% with retained sensory properties.

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2. Analysis

Graz University of Technology (Austria), Central Science Laboratory, York (United Kingdom), Queens’ University Belfast (United Kingdom), TÜBÝTAK-Marmara Research Center (Turkey),

Swedish Food Administration (Sweden), Universitat de Barcelona (Spain), Institute of Chemical Technology Prague (Czech Republic), Universitat de Barcelona (Spain), National Food Institute (Denmark),

Norwegian Institute of Public Health (Norway), Stockholm University (Sweden), Lund University (Sweden)

3.1. Method development Several experiments have been performed with the aim to optimize and validate analytical methods for heat-induced toxicants in different types of foods, of high accuracy and precision and suitable for use by producers and enforcement laboratories. Methodology for acrylamide analysis is now mature. The available LC-MS and GC-MS methods are capable of testing all the food types of interest down to the concentrations of interest. Accuracy is acceptable although precision could be improved. Method development and validation has been assisted by an understanding of the potential interferences in particular food types, and how to avoid them. To provide analytical data for intake estimates, the detection limit of the bromination-GC-MS method was improved to give a low (single-figure) µg/kg limit of detection. The improved method was applied to the analysis of UK total diet samples. The estimated dietary exposure to acrylamide was within, and at the low end of, estimates made in other countries. This Br-GC-MS method was found to be more variable than wanted in between-lab trials and an LC-MS/MS method was developed and validated. The development work included systematic studies on extraction conditions, optimised extract clean-up by SPE, and chromatographic column separation. The method was subjected to inter-laboratory validation under the auspice of JRC-irmm. Repeatability (r) figures were 4-9% with potato crisp samples and 3-8% with bakery products. The reproducibility (R) values were 9-13% and 5-12%, respectively. For evaluation of trueness, the study included a spiked potato powder (500 µg/kg) and a candidate reference material from crisp bread (980±90 µg/kg). The mean results for these were 485 µg/kg (RSDR=9%) and 950 µg/kg (RSDR=5%), respectively, indicating that the method bias is low or nonexistent. A variation of the method was developed for testing low acrylamide levels in baby food. The multimode step was omitted and proteins were precipitated by adding KAl(SO4)2 prior to SPE on ENV+. An LOQ of 0.5 µg/L was obtained. A new GC/HRTOF-MS analytical procedure enabling direct determination of acrylamide (no derivatisation) in various food matrices was validated. Also, a new LC-MS/MS method employing a novel sample preparation strategy was implemented. For quantification, either D3-acrylamide (GC/HRTOF-MS) or 13C3-acrylamide (LC-MS/MS) are employed. The accuracy of these methods is checked regularly via proficiency test scheme FAPAS®. Satisfactory z-scores were obtained: -0.3 (Baby Rusk Test Material, series 30 Round 9); 0.2 (Potato Crisps; 30:11), 0.4 (Breakfast Cereals; 30:14) and 1.5 (Crisp bread; 30:15). Performance characteristics are as follows:

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GC/HRTOF-MS LC-MS/MS LoD = 10-30 µg/kg, depending on matrix repeatability (RSD) = 2% (chips, ca 500 µg/kg)

LoD = 10-25 µg/kg depending on matrix repeatability (RSD) = 3% (chips, ca. 700 µg/kg)

Figure 5: SPME-GC-ITD/MS and SPME-GCxGC-TOF/MS methods for determination of heat-generated volatile compounds.

A method for the analysis of acrylamide by LC-MS/MS using an ion trap system with atmospheric pressure chemical ionization (APCI) was developed. A micro-emulsion electro-kinetic chromatography (MEEKC) method has been established suitable for acrylamide analysis at high levels in certain food types. Besides the direct analysis of acrylamide using LC-MS, a derivatisation method using 2-mercaptobenzoic acid was adopted that results in a substance with a higher molecular weight, which is more selective than the low molecular weight of acrylamide itself at 72. This method was used for the analysis of acrylamide in coffee and in model systems. The same derivatisation with mercaptobenzoic acid was used prior to a CZE analysis using FASI (field-amplified electrokinetic sample injection) as a pre-concentration on-line technique to increase sensitivity. Additionally a method for an intermediate compound was developed namely 3-amino-propionamide. This substance could be identified in a model system as an intermediate but it was not found during the roasting of coffee. A method for the determination of HMF (hydroxymethylfurfural) by GC-ITMS using N,O-bis(trimethylsylil) trifluoroacetamide as derivatising reagent has been developed and also an LC-MS method using an ion trap system with ESI as ionisation source. A method for the analysis of a metabolite HMFA (5-hydroxymethyl-furanoic acid) was developed. This metabolite is excreted via the kidney to the urine. The method was applied to the analysis of urine to estimate the exposure. The results suggest that the exposure is higher than calculated from the exposure data (food analysis). Therefore, it is necessary to analyse other foods and identify any additional source(s) of exposure. As part of method development for various heat-induced toxicants, different extraction techniques including solid phase extraction (SPE), solid phase micro-extraction (SPME) and pressurized solvent extraction have been explored to good effect. In response to the report on "masked" acrylamide, the pilot study of its formation in model system was initiated and possible precursors of “masked” acrylamide were identified.

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

Akrylamidm/z 71,036Hmotnostní okno: 0,02 Da

D3-m/z 74,056

í okno: 0,02 Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

m/z 71,036Hmotnostní okno: 0,02 Da

D3-Am/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

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8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

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7.43

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7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-m/z 74,056

í Da

TIC

Acrylamide

m/z 71.037

Mass window: 0.02 Da

-

m/z 74.056


6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

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7.43

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7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

/

D3-/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

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7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

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6.87

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7.43

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7.50 7.66

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7.72 8.027.857.79 7.98

8.21

8.448.37

D -/z 74,056

TIC

Acrylamide

m/z: 71.036

Mass winow: 0.02 Da

D3-Acrylamide

m/z: 74.056


6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-m/z 74,056

í okno: 0,02 Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-Am/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-m/z 74,056

í Da

TIC

Acrylamide

m/z 71.037


-

m/z 74.056


6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

/

D3-/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

D -/z 74,056

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-m/z 74,056

í okno: 0,02 Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

% 8.21

8.448.37


D3-m/z 74,056

í okno: 0,02 Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05

7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-Am/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time

6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-Am/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.49

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

6.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time

7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-m/z 74,056

í Da

TIC

Acrylamide

m/z 71.037


-

m/z 74.056


7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37


D3-m/z 74,056

í Da

TIC

Acrylamide

m/z 71.037


-

m/z 74.056


6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

/

D3-/z 74,056

í Da

TIC

7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

/

D3-/z 74,056

í Da

TIC

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

6.20 6.40 6.60 6.80 7.00 7.20 7.40 7.60 7.80 8.00 8.20 8.40Time0

100

%

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e3

0

100

%

0

100

%

2: TOF MS EI+ 71.036 0.02Da

9.91e37.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e3

7.12

6.20 6.496.26 6.667.28 7.687.73 8.04 8.338.29

2: TOF MS EI+ 74.056 0.02Da

5.25e37.12

2: TOF MS EI+ TIC

2.05e5

7.12

2: TOF MS EI+ TIC

2.05e57.33

6.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

D -/z 74,056

TIC

7.336.65

6.23

6.05 6.16

6.50

6.32

6.546.83

6.726.78

6.87

7.257.00

6.97 7.12

7.43

7.37

7.50 7.66

7.547.60

8.168.09

7.72 8.027.857.79 7.98

8.21

8.448.37

D -/z 74,056

TIC

Acrylamide

m/z: 71.036

Mass winow: 0.02 Da

D3-Acrylamide

m/z: 74.056


0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00Time0

100

%

0

100

%

crisps MRM of 4 Channels ES+ 75.05 > 58.35

2.41e52.66


1.27e52.66

0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00Time0

100

%

0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00Time0

100

%

0

100

%


2.41e5

0

100

%


2.41e52.66


1.27e5

2.66


1.27e52.66

13C3 – Acrylamidetransition m/z 75 → 58level: 1000 µg/kg

Acrylamidetransition m/z 72 → 55level: 650 µg/kg

13C3 – Acrylamidetransition m/z 75 → 58level: 1000 µg/kg

Acrylamidetransition m/z 72 → 55level: 650 µg/kg

GC/HRTOF-MS LC/MS-MS

of 77

3.2. Analytical results on heat-induced toxicants levels in foods The acrylamide levels in breast milk and Swedish baby food products were tested. The average levels for gruel, porridge and canned baby food, all ready to eat, were 1.4, 26, and 7.8 µg kg-1 respectively. For all breast milk samples except one, acrylamide was below the limit of quantification (0.5 µg kg-1). The mean intake between seven and twelve months of age was estimated to be about 0.5 µg/kg body weight/day. The acrylamide level in toasted bread was investigated in order to aid the estimation of intake from home-prepared foods. The amounts of acrylamide produced in the different breads by toasting were strongly related to the concentrations before toasting. There was hardly any acrylamide increase in the light toasted breads. The levels for medium toasting were about 2 to 5-times higher than the levels before toasting.

Table 3. Acrylamide concentrations in toasted bread (corrected for weight loss).

Weight loss Acrylamide Weight loss Acrylamide Weight loss Acrylamide Weight loss Acrylamide (%) (µg/kg) (%) (µg/kg) (%) (µg/kg) (%) (µg/kg)

no toasting 0 3,0 0 8,4 0 28,8 0 41,6

1 11 3,9 10 8,7 11 29,7 11 43,9

2 14 4,2 12 9,6 12 28,3 13 60,5

3 17 15,9 15 21,6 14 57,7 15 84,6

4 20 31,0 19 57,8 18 113,5 17 118,0

wheat Toasting

time

wheat wheat + rye wheat + rye

Analysis of acrylamide in foodstuffs (n=98) from Spanish markets and homemade processed foods has been performed. Similarly, a total of 227 data were collected for acrylamide contents in foods from the Turkish market, and 40 data for furan content. The survey for acrylamide included traditional Turkish foods and home-cooked foods. Moreover, French fries collected from fast food restaurants and French fries that were prepared by using frozen potato were also analysed. The French fries from fast food restaurants had the higher acrylamide content. Compared to other food groups, the traditional Turkish foods and bakery products contained lower levels of acrylamide. The survey was performed over two years. The same brands of products were monitored during this period. Although there is not enough data for proving it statistically, there were indications that acrylamide levels in some specific products, including baby biscuits, toasted bread and potato chips, decreased during this period. Typical results are given in Table 4.

Table 4. Data for acrylamide content in some food products sold in Turkey. Examples of Foods Acrylamide

(µg/kg) Examples of Foods Acrylamide

(µg/kg) Roasted almond 207-313 Fried vegetables (eggplant, pepper,

squash) <10

Pop corn 170 Pickled olives <10-139 Baby biscuits 36-613 Corn bread <10 Corn chips 198-835 Turkish coffee 227-300 Potato chips 59-2336 Instant coffee* 336 Bread (wheat) <10-79 Fried potato – home made 60-66 Toasted bread 58-205 Fried potato – frozen potato 72-76 Pilav (ready to eat rice) <10 Fried potato – fast food 375-428

of 77

The carcinogenic substance furan ’emerged’ as a heat-induced toxicant in foods during the planning and conduct of the HEATOX project. The issue originated from work by the US-FDA. Our studies made an important contribution in helping to understand aspects of the analysis of foods for furan – crucially the need to avoid strong heating during headspace analysis to avoid misleadingly high results. This provided acceptance criteria and so gave confidence in the analytical data on furan occurrence and concentrations that is being complied by EFSA. The headspace GC-MS method of testing for furan is straightforward and accurate. It can be used both by producers and official laboratories. Studies were then conducted to understand the chemical precursors, the mechanisms of formation, the nature of the foods affected by formation of furan, and the effect of normal consumer practice of heating and cooking before consumption. This work confirmed that there are different precursors and mechanisms of formation. On other furan compounds, HMF (hydroxymethylfurfural) was found at high levels in foods containing high amounts of carbohydrates that are stored for a long time or heated strongly. These included dried fruits, coffee, Madeira wine, caramel, balsamic vinegar, non alcoholic and alcoholic beverages. Comprehensive databases on (i) the reaction products from Maillard browning, and (ii) lipid oxidation products, were set-up and populated with occurrence data and with hazard indications derived from QSAR (quantitative structure-activity relationship) considerations. Similarly, a database for specified heat-induced toxicants (acrylamide, ethylcarbamate, furan, heterocyclic amines, hydroxymethylfurfural, polycyclic aromatic hydrocarbons and nitrosoamines) has been designed. Concentration levels of heat-induced toxicants in cooked food samples have been introduced in the database. Sample food and cooking data together with analytical information (sample treatment, identification and determination details) have been included in the database. As an example, in Table 5, there is the information about acrylamide in baby food products Table 5. Baby foods

Nº Samples

Nº NDSamples MIN MEDIAN MAX Units Determination

TechniqueExtraction and

clea-up procedures Ref.

Breast milk 4 0 0,25 0 0,65 µg/kg LC-MS/MS Precipitation of macromolecules (KAl (SO4)2)

49

Cereals 12 4 1 10 24 µg/kg LC-MS/MS S-L (H2O), L-L (AcOEt), SPE 49,80,125Infant formulas 67 64 - 10 - µg/kg LC-MS/MS S-L (H2O:MeOH 95:5), SPE 123,125Purées (Fruits, potatoes, vegetables, meat)

22 3 1 22 121 µg/kg LC-MS/MS S-L (H2O), SPE 49,125

Gruels and porridges 10 0 0,25 2 34 µg/kg LC-MS/MS S-L (H2O) 49Others (Biscuits, cakes, crackers, toasts)

10 0 19,9 118 1568,9 µg/kg LC-MS, LC-MS/MS S-L (H2O), L-L (AcOEt), Carrez, SPE

27,70,80,125

of 77

3.3. Analytical method for biomarkers

Figure 6. Biomarkers of AA for determination of short-term and long-term exposure

(Thomas Bjellaas - Doctoral Thesis)…

3.3.1. Identification of DNA-adducts in vivo An analytical method including the use of LC/MS has been developed for the detection of genotoxic compounds in a rodent bioassay. 2-methylfuran was detected as a possible genotoxic Maillard reaction product in this bioassay. More unknown genotoxins may be identified using this procedure. Mixtures of single amino acids and carbohydrates were suspended in phosphate buffer and heated to various temperatures for the production of Maillard reaction products as visualized by the colour formation. The Maillard reaction has been upscaled in order to produce sufficient material for rat feeding studies and a method for a fairly homogeneous incorporation of the Maillard reaction product into rat feed pellets has been developed. A study of the palatability in rats showed that two grams of Maillard product in 15 grams of rat feed was well tolerated after a short adaptation period.

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Rats were given feed with reaction products from different amino acids and after 5 days DNA was isolated from different organs. The DNA was hydrolysed and analysed by HPLC/MS. Chromatographic data of DNA from dosed animals were compared with data from control animals and differences, suggesting the presence of DNA-adducts, were detected by use of ”metabolite detect” computer software.

Figure 7. Extracted ion chromatograms m/z=232 of hydrolyzed intestinal DNA (upper) and a synthesised standard (lower).

At least two possible DNA adducts with mass 232 were detected by HPLC/MS analysis of hydrolysed DNA extracted from rodents following feeding with Maillard reaction products (Figure 7, upper panel). Analysis of the adducts by high resolution TOF-MS gave exact masses of 232.08345 for both adducts which is in accordance with the molecular formula C10H10N5O2. The molecular formula suggested a guanine adduct to which C5H6O2 had been added followed by elimination of water. A possible candidate to one of the adducts 4-oxo-2-pentenal was synthesised by oxidation of the known Maillard reaction product, 2-methylfuran with m-chloro-perbenzoic acid. Reaction of 4-oxo-2-pentenal with deoxyguanosine gave a product which after acid hydrolysis had identical retention time and mass as the first eluting adduct found in rodent DNA (Figure 7, lower panel). 3.3.2. Acrylamide metabolites as biomarkers of exposure Exposure to acrylamide from food has been estimated using records of food consumption combined with knowledge on acrylamide content of various foods. However, these intake estimates needs to be validated against biological markers of exposure. The concentration of haemoglobin adducts of acrylamide reflects the internal dose of acrylamide during the last 3-4 months, while we had no markers for short term intake of acrylamide. The bioavailability of acrylamide from food is an important issue which was addressed in experiments on mice. In one study, 53 persons answered a food frequency questionnaire about the daily food intake and also had answered a 24 h food recall. Blood and 24h urine samples were collected. In another study, mice were kept in metabolic cages and were exposed to crisp bread with different concentrations of acrylamide, subcutaneous injections of corresponding doses of acrylamide, including 14C-labeled acrylamide.

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Acrylamide and its oxidation product glycidamide are conjugated to glutathione and these conjugates are stepwise converted into N-mercapturic acids which are excreted into urine fairly rapidly. The mercapturic acid metabolites including 2H-labelled standards as well as an external standard were synthesised and characterised (Figure 8).

Figure 8. Structures of standards used in the study.

An analytical method of determination was developed based on solid-phase extraction of urine followed by LC-MS/MS separation and quantification (Figure 9). The method of analysis was applied in a pilot study of six persons. Fasting caused a 50 percent reduction in excretion of mercapturic acid metabolites clearly showing that food is an important source. Smoking was associated with a high excretion of mercapturic acids.

Figure 9. HPLC-MS/MS chromatogram of (A) a spiked urine sample at 40 µg/L for MA-AA and MA-GA3, (B) urinary metabolites from a non-smoker and (C) urinary metabolites from a smoker. Peak intensities are set relative to the spiked urinary sample. The grey shaded peak at 8.2 min might possibly be the sulfoxide analogue of MA-AA.

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It was shown that excretion of urinary mercapturic acid metabolites was related to food intake, the biomarker responded quickly to food with a high content of acrylamide, e.g. potato crisps (Figure 10).

Figure 10. The total amount of AA found as urinary metabolites in consecutive urine samples in a participant who reported intake of potato crisps during the evening.

On the assumption that 55% of the acrylamide dose is excreted as mercapturic acids, the average dose in the 53 persons was 30 µg/day. Excretion correlated with estimated recent intake, but not with the total estimated amount during the last 24 hrs. Multiple regression analysis showed that the intake of aspartic acid (including also asparagine), starch and coffee were significant predictors of AA metabolite excretion. The method of urinary excretion could also be used for the identification of particular foods contributing to acrylamide exposure. Acrylamide haemoglobin (Hb) adducts were determined in the same study subjects. The level decreased with age, possibly explained by a lower intake of foods with high acrylamide content, such as potato chips and snacks, in middle aged and older persons. A 3.5-fold higher adduct concentration was found in smokers in comparison with non-smokers. No correlation was found between Hb adduct levels and total dietary acrylamide intake estimated from reported food consumptions and levels of acrylamide in foods. After adjusting for age significant predictors for Hb adduct formation were intake of chips, snacks and crisp bread. Haemoglobin adduct concentrations did not correlate with urinary excretion of mercapturic acid derivatives in the same subjects.

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In mice exposed systemically to 14C-acrylamide, about 92% of the radioactivity was excreted into urine during the following 48 hrs. There were linear dose response curves both for acrylamide from crisp bread and systemically given (11). The bioavailability of AA from crisp bread was about 100%.

Figure 11. A) The linear relationship between the amount of acrylamide (AA) ingested from crisp bread and the amount of AA recovered as urinary metabolites. B) The percentage of dose recovered in urine for total and the different urinary biomarkers MA-AA, MA-GA3, MA-GA2 and GA

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3. Hazard characterisation Swedish Food Administration (Sweden), Lund University (Sweden), Stockholm University (Sweden), German Institute for Human Nutrition (Germany), Norwegian Institute of Public Health (Norway),

University of Zürich (Switzerland)

4.1. In-vivo genotoxicity and carcinogenicity In absence of data on the effects of a chemical on humans it is often necessary to extrapolate the effect from high doses given to animals to lower dose regions for exposed humans. Therefore when the effect of very low experimental doses can be determined the extrapolation is more reliable and makes the risk quantification more reliable. The work has been concentrated around the uptake and the effect in the low dose region of acrylamide and other chemicals produced when heating foodstuffs. To clarify the shape of the dose response curve in the low dose region the biomarkers for DNA adducts, haemoglobin adducts for acrylamide and glycidamide, and the micronucleus assay (two-lazer-beam flow cytometer) were used. The mice (35) were given by gavages 14 different doses (control included) of acrylamide. The doses were ranged from 0 to 60.0 mg/kg body weight. Data from the measurement of haemoglobin adducts, Figure 12, and from the micronucleus (MN) assay, Figure 13, are given below. It is clearly shown that there is a dose related increase of the frequency of MN (fMNPCE). The fMNPCE levels for the different mice are about the same as after intraperitoneal injection. The dose response is linear. Also the analysis of haemoglobin adducts showed a linear dose related increase. However, the lowest level of the quantification of haemoglobin adducts was about 1 mg/kg.

Figure 12. Measurement of hemoglobine adducts Figure 13. Micronucleus assay

Results from studies using accelerated mass spectrometry (AMS) show that glycidamide binds to DNA in mice at a dose of about 1.5 µg/kg body weight. In a human study where twenty-four healthy persons eating either a) boiled (max 100 ºC) and fresh food or b) strongly heated (fried) and fresh food, there was a significant difference between the groups regarding the frequency of micronucleated young erythrocytes. From each of the subjects blood samples were drawn twice, before and after an “eating period” of four days. Blood samples were analysed for both the frequency of micronucleus (MN) in young erythrocytes and the haemoglobin adduct levels of acrylamide and glycidamide.

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Figure 14. frequency of micronucleated young erythrocytes in twenty-four healthy persons eating either a) boiled (max 100ºC) and fresh food or b) strongly heated (fried) and fresh food …

There are different possibilities to increase the sensitivity of the micronucleus assay in human erythrocytes; one way is to e.g. discriminate between clastogenic and aneugenic effects. This can be done by a fluorescent in situ hybridisation (FISH) of the DNA in the micronuclei. Another way (without FISH) is to optimize the fixation method, in order to better define the cell population (very young erythrocytes) when analysing the cells in the flow cytometer. This latter option was used as a new refined method when the human study was done. Also other chemicals than acrylamide have been studied upon the potency to increase the frequency of micronuclei, for example furan, metylfuran, HMF, its metabolite SMF, phytosterol oxides, metylacrylamide and also extracts of potato chips. Among these chemicals only SMF showed a positive effect in the in vivo micronucleus assay in mice. A comparison of the genotoxicity of acrylamide and three heterocyclic amines (PhIP, IQ, and MeIQx) was made. In the case of PhIP, we found a significant dose-response relationship, while MeIQx and IQ did not give an increased fMPCE level. A comparison between the heterocyclic amines and acrylamide revealed that the slope of the dose-response curve is about ten times steeper for PhIP than acrylamide. In spite of this, acrylamide probably constitutes a higher human risk than heterocyclic amines since the intake of acrylamide is about a hundred to a thousand fold higher. In conclusion, the results points to an increased risk already at the lowest doses. In the human study the acrylamide dose was about a thousand times lower than the lowest effective doses in earlier studies in mice. It is likely that the explanation of the increase of chromosome instability (micronuclei) is something else than acrylamide and it is possible that the high heating temperature of foodstuffs is responsible for the production of chemicals other than acrylamide inducing a higher frequency of micronuclei.

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4.1.1. Data on the relationship between internal dose and estimated intake of acrylamide in blood samples from a biobank Associations between estimated acrylamide-intake, based on self-reported data on usual food consumption combined with data on published acrylamide content in foods, and Hb adducts were examined with linear regression and correlation analysis. A sample from the Malmö Diet and Cancer cohort was selected to obtain a large variation in acrylamide-intake. The estimated acrylamide intakes ranged from 3.5 to 190 µg/day, with a median of 45 µg/day. In randomly selected individuals the estimated intake was 28 µg/day. The estimated intake thus varied with a factor of ca 50. In contrast to this, the internal doses as measured by acrylamide adduct levels in Hb varied with a factor of 5 in the non-smoking group (see 15). Hb adduct levels are expected to give a more exact estimate of the intake than the estimates from questionnaires. Positive partial correlations between dietary acrylamide-estimates and Hb adducts were seen in non-smoking men but not in non-smoking women. In conclusions, two results from this study cast doubt on the validity of dietary intake estimates used in cancer epidemiology: a) The lack of correlations between adduct levels and intake estimates in non-smoking women, b) The much lower variation in internal doses compared to estimated intakes from food frequency questionnaires.

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We studied some non-acrylamide compounds that form when carbohydrates are heated, in particular 5-hydroxymethylfurfural (HMF) and related compounds. These compounds are ingested at extremely high levels (1000 times higher than acrylamide) and it has been reported that rodent sulfotransferases (SULTs) can covert HMF in vitro to a mutagen, 5-sulphooxymethylfurfural (SMF). We wanted to find out whether human SULTs can mediate this reaction in vitro and in vivo and to study resulting adverse effects . The objectives included the development of biomarkers for this activation pathway in animals and humans. Our results may be summarised as follows: A major human SULT form (SULT1A1) was very efficient in converting HMF to SMF. In rat and mouse, only minor SULT forms showed high sulfation activity towards HMF. Therefore, mice and rats may be poor models for humans when studying SULT-mediated toxicity of HMF and congeners. SMF was identified as a urinary HMF metabolite in a mouse line engineered for expression of human SULT1A1, but not in wild-type mice, corroborating that human SULT1A1 may be a risk factor in exposures to HMF.

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SMF, unlike HMF, formed DNA adducts in cell-free systems, was mutagenic in conventional bacterial and mammalian cell test systems, and was strongly nephrotoxic in mice (Figure 16). HMF and other hydroxymethylfuran congeners present in foods were mutagenic in bacteria and mammalian cells engineered for expression of human SULT1A1. Whereas HMF was only mutagenic when SULT1A1 was expressed, some analogues were also mutagenic in control V79 cells (but not in control bacteria) suggesting the existence of additional bioactivation mechanisms in mammalian cells (e.g. mediated by unidentified kinases). Externally added furfuryl sulphate (unlike SMF) was only cytotoxic, but not mutagenic, to bacterial and mammalian cells. However, when furfuryl sulphate was generated by SULT from its alcohol within target cells, it was strongly mutagenic. This difference in biological activity between internal and external furfuryl sulphate might be due to its charge and short half-life time preventing permeation of the cell membranes. Adenine was identified as the major nucleobase forming DNA adducts with SMF in cell-free systems. This and other adducts were not detected in target cells and tissues of SMF despite increased mutant frequencies and massive histopathological alterations. Unfortunately we did not succeed developing sufficiently powerful enrichment methods for adducts, as required for their demonstration in cells and tissues. Moreover, the presence of the carbonyl group in 2-postion of HMF/SMF, expected to be still present in the adducts, complicated the analysis, as it might form cross-links with DNA and/or other cellular constituents. We have demonstrated that major human SULTs, unlike major rat and mouse SULTs, efficiently convert HMF to SMF, and that SMF forms DNA adducts in cell-free systems, is mutagenic in bacterial and mammalian cells in vitro, and is strongly nephrotoxic in mice. The negative result of HMF in conventional in vitro systems is due to a inappropriate metabolic situation, as HMF is positive in the same tests after expression of human SULT1A1 at levels occurring in human tissues. Likewise, the low biological activity of HMF in laboratory animals should be regarded with caution, as these models do not truly reflect the human sulfation capacity. A similar situation was found with other hydroxymethylfuran congeners present at high levels in food, such as furfuryl alcohol.

Figure 16. Kidney of an SMF-treated mouse. Note that the proximal tubules were completely destroyed, whereas the glomeruli were damaged much less. The right panel shows abundant proteinaceous casts.

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4.2. Tumorigenicity Two experiments on intestinal tumourigenesis of acrylamide and glycidamide in Min/+ mice and their wt litter mates have been completed. In the first experiment the animals were exposed to acrylamide or glycidamide (10 or 50 mg/kg bw) week 1 and 2 after birth, and in the second experiment they were exposed to acrylamide or glycidamide one week before birth to the dam, alone or in combination with exposure of the pups at week 1 and 2 after birth (50 mg/kg bw). Min/+ mice and wild type mice have been treated postnatal with subcutaneous injections of 5-hydroxy-methyl-furfural (HMF) or its reactive metabolite 5-sulfoxy-methyl-furfural (SMF). Eight weeks old female Min/+ and wt mice were exposed to glycidamide at the dose 50 mg/kg dissolved in 0.9% NaCl or to NaCl-solution alone for 6 and 24 hours. RNA was isolated from the epithelium and cDNA was made by in vitro transcription of total RNA, and Affymetrix GeneChipswere used for the expression analysis. A weak tumourigenic activity in the intestine of Min/+ mice and their wt litter mates was observed after exposure to acrylamide or glycidamide. The weak tumourigenicity observed was probably related to the perinatal exposure since such effects never previously have been observed in rodents exposed later in life. SMF significantly increased the number of adenomas in the small intestine and the number ACF in the colon of Min/+ mice. HMF did also show a tendency to increase the number of lesions. Gene expression profiles showed a high degree of variability and only one gene was significantly up-regulated by glycidamide in wt mice exposed for 6 h (metallothionein-2, 3.27-fold, adjusted p<0.05). No other genes were significantly regulated in each of the treatment. We investigated differences in gene expression between 6 and 24 hours, expecting different expression of genes involved in circadian exercises. We had from 75 to 1790 significantly regulated genes (adjusted p < 0.05) in each of the four data sets, and indeed, genes involved in circadian rhythms came out as a major functional cluster. 4.2.1. Reproductive toxicity Isolated mouse testicular cells and lymphocytes (primary cells) from BigBlueTM C57BL/6 male mice, either mOgg1+/+ or mOgg1-/- (a transgenic mouse strain deficient in oxidated lesion repair) were used. Peripherial blood lymphocytes from humans were also used. The alkaline single cell gel electrophoresis (comet assay) has been used to measure levels of DNA lesions as single strand breaks and alkali labile sites (baseless sites) after in vitro exposure of acrylamide (AA) and glycidamide (GA) in mouse and human cells (see above). To further study and characterise the DNA lesions formed, a modified comet assay was used. The modified assay includes a treatment step in which cellular DNA is subjected to an endogenous DNA repair enzyme that recognises and cleaves certain DNA lesions thereby converting them to single strand breaks, so that they are measurable in the comet assay. The CYP2E1 AA metabolite glycidamide was shown to be genotoxic in mouse testicular cells and lymphocytes in vitro, at concentrations similar to those producing genotoxicity in human lymphocytes. The parent compound AA was as expected very inactive. The DNA lesions induced by GA exposure are recognised by repair enzyme which dramatically increases the lesion levels, implicating that human testicular cells might be particularly susceptible to the active metabolite of acrylamide.

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4.2.2. Neurotoxicity Neonatal mice were exposed to different low doses of acrylamide at an age of 10 days. This study indicates that neonatal exposure to acrylamide causes developmental neurotoxic effects that are persistent and also get worse with age. These disturbances are both dose-response and time-response related. The spontaneous motor behavior data showed a disruption of habituation in animals exposed to acrylamide. Habituation, here defined as a decrease in the locomotion, rearing, and total activity variables in response to the diminishing novelty of the test chamber over a 60 min. period, was displayed in the control animals, but the animals exposed to acrylamide were clearly hypoactive during the beginning of the 60 min. period, while towards the end of the test period they were hyperactive. Furthermore, the lowest dose not causing any developmental neurotoxic effects (NOEL) on spontaneous behaviour is 0.1 mg acrylamide/kg body weight. Acrylamide is tumorigenic at high doses in rodents and has been classified as a probable human carcinogen. However, cancer risk projections in the general population remain problematic because the molecular pathogenesis of acrylamide at the low level of dietary uptake is not understood. In particular, the question of how non-genotoxic (epigenetic) responses such as changes of gene expression may modulate the known genotoxicity of acrylamide has never been examined. To close this gap of knowledge, we used high-density DNA microarrays and real-time PCR validations to determine genome-wide transcriptional profiles induced by acrylamide and glycidamide, the oxidative metabolite of acrylamide. Human MCF7 cells (derived from the mammary epithelium) and human CaCo-2 cells (derived from the intestinal epithelium) were exposed to acrylamide or glycidamide. After 24-h exposures, the cells were collected, RNA was extracted and processed for genome-wide expression profiling on Affymetrix microarrays that display the sequences of 47’000 human transcripts. Glycidamide induced gene expression changes of much higher amplitude than acrylamide itself. We identified > 100 human transcripts, coding for proteins with known biological functions that are significantly up- or down-regulated by exposure to glycidamide. The expression profiles resulting from glycidamide treatment are characterized by the dose-dependent up-regulation of cytoprotective factors including the glutathione system, enzymes that reduce toxic carbonyl compounds as well as diverse antioxidants. Low-dose experiments indicate that the transcript coding for epoxide hydrolase-1 represents the most sensitive biomarker for glycidamide exposure. At higher concentrations, glycidamide induces typical markers of tumour progression such as steroid hormone activators, positive regulators of nuclear factor-kB, growth stimulators and apoptosis inhibitors. Concomitantly, transcripts coding for growth suppressors and cell adhesion molecules are down-regulated. Many of these transcriptional changes have been validated by quantitative real-time reverse-transcriptase PCR assays in both MCF7 and CaCo-2 cells. In the case of epoxide hydrolase-1, a significant transcriptional induction was observed at a glycidamide concentration of as low as 0.1 µM. This is a glycidamide level that may be reached transiently in the blood or tissue of humans after dietary uptake of acrylamide. The micromolar doses of glycidamide that induce many other transcriptional responses might seem too high to be relevant for human exposure. However, it cannot be excluded that the sensitivity of target cells depends on the capacity of their detoxification systems and, considering that the uptake of acrylamide is normally accompanied by other toxic food constituents that may saturate these cytoprotective systems, adverse effects may occur at lower concentrations than those expected from threshold experiments performed with a single compound. This functional genomic study revealed that acrylamide and, in particular, glycidamide exert a plethora of transcriptional effects that should be included in risk assessment studies. We propose to implement a threshold level that prevents the induction of potentially hazardous transcriptional responses mediated by acrylamide or glycidamide in target tissues such as the mammary gland or the intestinal epithelium.

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The results of the in vitro study suggest that low-dose responses, primarily the induction of epoxide hydrolase-1, exert beneficial effects by promoting inactivation of the toxicant. Other detoxification processes observed at the low-dose glycidamide level of 0.1 µM include its inactivation through conjugation with glutathione (see Figure 17). Adverse effects, such as for example the down-regulation of cell-adhesion molecules (CDH12, ANXA1, ITGB6), the up-regulation of factors that lead to dysregulation of steroid hormone synthesis (AKR1C2, AKR1C3), or factors that promote cell survival and cell cycle progression (NF-kB, RRM2, Cyclin-1, MAPK12) are detected at concentrations above the range that appears to be relevant for human populations. An induction of the DNA repair enzyme EME1, indicative of the formation of DNA damage, is also observed in the upper level of the tested dose range. Thus, the main implication of these findings for risk assessment is that transcriptional signatures associated with DNA damage or tumour cell progression may be expected only at doses that exceed the range of ordinary dietary exposure.

Figure 17. Summary of the main findings from the transcriptomic profiling of glycidamide effects in human cells. Beneficial responses are observed at the low-dose level, potentially hazardous effects are detected at high glycidamide concentrations. Abbreviations: EPXH1, epoxide hydrolase-1; GSH, glutathione; FHO3, formin homology-3; CDH12, cadherin-12; NF-kB, nuclear factor-kB; RRM2, ribonucleotide reductase polypeptide-M2; AKR1C2, aldo-keto reductase-C2; AKR1C3, aldo-keto reductase-C3; EME1, essential meiotic endonuclease-1; ANXA1, annexin-A1; ITGB6, integrin beta-6; MAPK12, mitogen-activated protein kinase-12.

Consequences for risk assessment: these findings indicate that the standard procedure of extrapolating from high-dose effects in rodents or other experimental systems to the very low-dose level of human exposure would lead to an overestimation of the toxicological risk, thus exaggerating the health hazards resulting from the presence of acrylamide in food and its metabolic conversion to glycidamide in the human body.

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4. Exposure Assessment RIKILT Institute of Food Safety (The Netherlands), Swedish Food Administration (Sweden),

Stockholm University (Sweden), National Food Institute (Denmark)

5.1. Monte Carlo Risk Assessment For the work on the exposure assessment the Monte Carlo Risk Assessment (MCRA) software has been used. MCRA was developed in other projects, but adapted slightly for HEATOX purposes e.g. an additional option has been developed to study the effect of brand loyalty on exposure. 5.1.1. Dietary exposure assessment The long-term exposure of the Dutch population has been calculated, using the EU database on acrylamide concentrations. Exposure of the total population ranged from 0.4-1.8 µg/kg bw/day (P50-P99), exposure of children aged 1-6 years ranged from 1.0-2.4 µg.kg bw/day. Also a preliminary calculation of the exposure to furan has been made. With the data available from US FDA in 2004, the exposure to furan of the Dutch population was estimated at 0.25-0.87 µg/kg bw/day (P50-P97.5). With the purpose to validate the MCRA software duplicate diets of infants have been collected and analysed. Since only one of the diets contained a quantifiable amount of acrylamide it was not possible to use this study for validation.

Figure 18. Contribution of food groups to acrylamide exposure in different countries for different age groups.

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5.1.2. Scenario-analysis of effect of new processing techniques Within the HEATOX-project research is performed to develop new or adapted processing techniques to reduce acrylamide (AA) levels in the following foods; bread, crispbread, coffee, crisps and French fries. These reduced acrylamide levels were applied in scenario analyses with the aim to give insight in the theoretical reduction in AA exposure. The scenario-analysis showed that the reduction of the AA levels in single food groups had small effects on the exposure. When reductions of all food groups are applied in one scenario ‘total’ this resulted in a reduction of the exposure of the total Dutch population by approximately 40%, to 0.3 µg/kg bw/ day (P50) and to 1.2 µg/kg bw/ day (P99.9 ).For children the exposure decreased to 0.6 µg/kg bw/ day. Figure 19 shows the exposure expressed as Margin of Exposure per scenario for the upper 50% of the population. It should be noted that the new processing techniques which were applied in the scenarios were developed on lab scale and are not yet applied on industrial scale; thus the shown reduction is only potential. 5.1.3. Brand loyalty With the developed brand loyalty model the issue of brand loyalty in the scenario-analysis of acrylamide exposure assessment. The model was applied on the case of potato crisps, for which data on levels of different brands was used from a German data source (www.foodwatch.de), and market shares and brand loyalty were simulated. These calculations showed that brand loyalty has influence on the higher exposure levels and that brands with high acrylamide levels can increase exposure levels depending on the market share.

Figure 19. Distribution of Margin of Exposure (MoE) of the acrylamide reduction scenarios over the percentiles.

5.1.4. Home cooking Only limited data was available on the levels in home-cooked foods and no information was available on the amount of home-cooked foods in the diet. The levels reported for home-cooked foods are mostly within the range of levels in the EU-database, thus the intake from home-cooked foods is at least partly covered by intake calculated with the EU-database. However, from the first estimates we may conclude that home-cooking can have (a probably small) influence on the acrylamide exposure, both in increasing and decreasing exposure, therefore it is important that consumers follow the advice not to overcook their food and follow the available cooking guidelines. From the experiments known today it appears that by following the cooking guidelines acrylamide intake form home-cooked foods can be kept as low as possible.

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5.1.5. Intake in British and Dutch infants A comparison has been made between the acrylamide exposure of Dutch and UK infants using the same exposure assessments methodology. It appears that UK infants have a two times higher intake based on the difference in their diet of acrylamide containing food items. A standardized international methodology for exposure assessment is recommendable.

5.2. Training on probabilistic exposure assessment A training on probabilistic modelling of exposure to acrylamide was organised for European National Food Safety Authorities, including EFSA. The training was aimed at those working at the authorities and who are involved in risk assessment of acrylamide or other heat-toxicant research. In the training participants learned about the probabilistic approach and learned to work with the Monte Carlo Risk Assessment software in relation to acrylamide. Specific acrylamide issues were discussed and studied through exercises, like the grouping of foods, variability in the acrylamide levels, effect of mitigation measures and the possibility to study and how to quantify these aspects with MCRA. The training was very well received, and all participants were able to use the Monte Carlo Risk Assessment (MCRA) software for probabilistic exposure calculations. The calculation performed on acrylamide exposure gave a good estimate of the exposure of the Dutch population. Work on brand loyalty and home-processing showed that these issues may influence exposure, especially the exposure of the high-end consumers may be increased. The effectiveness of different mitigation possibilities to reduce acrylamide exposure levels has been studied using probabilistic models. Some of the mitigation measures showed a significant reduction on a particular food item, but overall the effect on exposure levels was limited. A significant reduction in exposure can only be received when mitigation measures are applied in a variety of products. Comparing acrylamide levels of different countries has been done, and the methodology of how to perform probabilistic intake calculations and scenario studies regarding the effectiveness of different mitigation options has been trained to different European National Food Authorities. This was received very well, and has resulted in new proposals for future cooperation.

5.3. Estimation of breast cancer risk from exposure to heat-induced toxicants An LC/MS/MS method for the analysis of haemoglobin adducts of acrylamide (AA) and glycidamide (GA) was established. The procedure involves isolation of reed blood cells, lysis and isolation of haemoglobin. Normal and adducted N-terminal valine amino acids are liberated by a modified Edman degradation followed by column purification. Normal and adducted valine residues are separated by gradient HPLC with MS/MS detection. The LC/MS/MS method has been optimised and the use of deutorated internal standards (Figure 20) has been included in the analytical procedure. Standard curves for AA-Val-PTH are linear from 0.3 pmol/ml to 28 pmol/ml and for GA-Val-PTH (two diasteriomers) standard curves are linear from 0.8 pmol/ml to 48 pmol/ml.

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Different sample purification methods were evaluated, including solid phase extraction (SPE) with different columns, columns with diatomaceous earth and liquid/liquid extractions. A useful and reproducible method using SPE was established.

Figure 20. Chromatograms showing GA-Val-PTH and AA-Val-PTH (upper) as well as the deutorated internal standards (lower)

From 1993 to 1997 blood samples were collected from approx 30.000 women in Denmark. Since then about 430 postmenopausal women had been diagnosed with breast cancer. Globin was purified from the 430 blood samples from the cancer patients as well from the same number of controls. The globin samples were analysed for content of AA-ValPTH and GA-Val-PTH by the LC/MS/MS method. Results of the unadjusted statistical analysis showed that neither acrylamide [IRR (95% CI) 1.01 (0.65-1.57)] nor glycidamide [IRR (95% CI) 0.79 (0.45-1.38)] concentrations were found to be associated with breast cancer incidence. Inclusion of potential confounding factors in the model did not have clear effects on the association between glycidamide and breast cancer, whereas the association between acrylamide and breast cancer was strengthened after inclusion of information on smoking at blood sampling [IRR (95% CI) 1.43 (0.75-2.71)], further inclusion of former smoking, duration of smoking and amount of tobacco currently smoked [IRR (95% CI) 1.70 (0.85-3.38)], as well as additional inclusion of baseline risk factors for breast cancer [IRR (95% CI) 2.04 (1.00-4.18)]. When the concentrations of glycidamide and acrylamide were evaluated with estrogen receptor (ER) specific breast cancer as the outcome, all associations was found to be strongest with regard to ER+ breast cancer. In the fully adjusted model, women with the highest concentrations of acrylamide adducts were found to be at almost 3-times increased risk of ER+ breast cancer when compared to women with the lowest concentrations (p=0.01).

5 6 7 8 9 10 11 12 13 14 Time [min]0.0

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5.4. Measurement of internal doses of acrylamide and glycidamide – a tool for exposure assessment and risk assessment 5.4.1. Haemoglobin adducts as a measure of internal dose Doses in blood of reactive compounds, like acrylamide and its metabolite glycidamide, could be inferred from levels of their respective adducts to haemoglobin (Hb). Hb adduct levels from acrylamide and glycidamide were quantified as a measure of internal doses in samples from humans and animals in different studies. Preparation of samples and analysis of adducts were performed according to an established method (the N-alkyl Edman procedure). 5.4.2. Bioavailability of acrylamide in food The bioavailability of acrylamide from two different kinds of diets, one containing potato and one containing fibre-rich bread, has been studied in mice. No difference in the bioavailability of acrylamide between the different types of diets was observed. Furthermore, it was shown that the “extra acrylamide”, obtained during alkaline extraction of the fibre-rich diet, does not correspond to bioavailable acrylamide. 5.4.3. Endogenous formation of acrylamide Internal dose of acrylamide in blood were measured in free-living animals with assumed no dietary intake of acrylamide; common hare, red deer, roe deer, red fox and wild boar. In all these species the adduct levels were shown to be very low (below or close to the limit of quantification). The conclusion is that there is no or very low endogenous production of acrylamide in mammals, and that possible endogenous formation gives insignificant contribution in humans compared to intake through diet. 5.4.4. Inter- and intra-variation of internal doses in humans Inter-individual variation in acrylamide- and glycidamide doses was studied in blood samples from 142 smoking and non-smoking individuals in the general population. The acrylamide adduct levels varied between 0.02 to 0.1 nmol/g globin (a factor of 5) in non-smokers. In smokers the range of the acrylamide-adduct level was 0.03 - 0.43 nmol/g globin. The variation in glycidamide doses was slightly higher. The results indicate a non-linearity in the internal doses of glycidamide at low exposure doses of acrylamide. This observation has to be more carefully investigated.

Non-smokers

0

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0,04

0,06

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Figure 21. Variation in acrylamide-adduct levels in Hb between non-smoking individuals (n=70).

The intra-individual variation of adduct levels from acrylamide was studied in blood samples collected from 15 non-smokers every second month during 1.5 years. There was an intra-individual variation in the acrylamide adduct levels in Hb over time, assumed to reflect a variation in background exposure. This variation was, however, lower than the inter-individual variation.

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5.4.5. Relationship between internal dose and estimated acrylamide intake from food The average acrylamide-intake, estimated from food frequency questionnaires (FFQ), varied nearly with a factor of 50 between the donors of the blood bank samples mentioned above. This means nearly 10 times larger individual variation compared to the measured internal doses (vary with a factor of 5). This illustrates the difficulties with estimation of intakes from FFQ. In two human studies volunteers were given controlled intake of food with known acrylamide content. Levels of Hb-adducts from acrylamide were compared with estimated intakes. In each study, about 20 individuals, smokers and non-smokers were having a 28- respectively 5-days period of controlled intake of acrylamide-rich diet (in average 3 and 11 µg acrylamide/kg and day, respectively). In blood samples, collected before and after the exposure periods, Hb-adducts from acrylamide was measured. Both studies clearly showed increased Hb-adduct levels corresponding to the acrylamide intake in both non-smokers and smokers.

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6 Risk assessment and risk communication Swedish Food Administration (Sweden), Norwegian Institute of Public Health (Norway), Stockholm University (Sweden),

National Food Institute (Denmark), RIKILT Institute of Food Safety (The Netherlands), BEUC - European Consumers' Organisation (Belgium), National Veterinary Institute (Norway), Lund University (Sweden)

6.1. Risk assessment 6.1.1. Models for extrapolation of cancer risk According to a relative cancer risk model (applied/developed by the partner) the species extrapolation from cancer tests in rodents should be based on internal doses of the ultimate genotoxic agent in the test species and in humans. Internal doses are obtained from Hb-adduct levels.

Figure 22. Important corner-stones in the approach for cancer risk estimation.

Rats were exposed to acrylamide in drinking water in an experiment simulating the conditions used in published cancer tests, and after one week blood samples were collected. In other studies blood samples from humans with background exposure to acrylamide (samples from blood banks), or from humans with increased exposure to acrylamide (experiments with dietary exposure), were studied. Hb-adduct levels from acrylamide and glycidamide were measured. Complementary experiments have been done to allow calculations of internal doses from the measured adduct levels in rodents and humans. Results with non-linearity of the internal doses of glycidamide at low exposure doses of acrylamide were strongly indicated in the human experimental studies. Taken together, the results indicate a relatively lower dose of glycidamide at low acrylamide doses. However, as these low exposure doses are associated with a larger measurement error, the indicated results of non-linearity has to be further strengthened. The result, if true, considerably complicates the cancer risk estimation. Therefore, further verifying measurements are required. 6.1.2. Final Risk Characterisation The work performed in HEATOX together with results obtained outside the project are summarised and commented below. For non-carcinogenic effects of acrylamide, the most sensitive endpoint is neurotoxicity with a NOAEL (No Observable Adverse Effect Level) in experimental animals of 0.2 mg/kg bw/day. The margins of safety relative the NOAEL for the average and high exposure were 200 and 50, respectively. The safety margins for other effects were larger. Comparing estimated NOAEL for neurotoxicity in humans of 10-40 µg/kg bw/day with exposure from food resulted in a safety margin in the range of 2.5 to 40. Given these margins any risks of neurotoxic and other non-carcinogenic effects following dietary acrylamide exposure are likely to be very small.

Adduct level in blood

Internal dose and uptake

Cancer risk

ReactivityPharmaco-kinetics

Genotoxic potencyCancer risk model

Adduct level in blood

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There are obvious problems with epidemiological studies, particularly those addressing low level exposure through food in the general population using Food Frequency Questionnaires (FFQ). First of all none of the studies published so far were designed for investigating the carcinogenic risk of acrylamide. There is no validation of the exposure assessments, and in some of them all dietary sources of acrylamide had not been taken into account. A comparison of intake estimates based on FFQ with that of acrylamide haemoglobin adducts showed large overlaps; a major problem is the correct classification according to exposure. Misclassifications would strongly weaken a hypothetical association between acrylamide exposure and cancer. The next limitation is the ability of the dietary studies, and even the occupational cohort study, to detect the small increases in risk expected from the lowest to the highest exposure groups. Indeed, neither the epidemiologic study nor the studies on dietary exposure to acrylamide did have the power to detect small increases in cancer risk and should be regarded as non-positive and not negative studies. Another issue is the mechanism of cancer induction by acrylamide. There are examples of chemicals that can induce heritable epigenetic modifications to DNA and thereby change gene expression without altering DNA sequence. So far there are no data to prove that epigenetic changes are important in acrylamide carcinogenesis although, for example, tumour location in animals might indicate a hormonal component in the aetiology. All data reported support a genotoxic mechanism, mediated by glycidamide, for acrylamide carcinogenesis. However, this needs further investigations. JECFA compared exposure estimates of 1 and 4 µg/kg bw/day with the BMDL10 of 30 mg/kg bw/day (BMDL10=Bench Mark Dose Lower confidence limit for 10% increased incidence) and obtained margins of exposure (MOEs) of 300 and 75. JECFA considered these exposure margins to be low for a compound being both carcinogenic and genotoxic and that they indicate a human health concern (WHO, 2005). The Scientific Committee of EFSA (EFSA, 2005) also endorsing a MOE approach for compounds that are both carcinogenic and genotoxic is of the view that a MOE of 10,000 or larger would be of low concern from a public health point of view and might be considered as a low priority for risk management actions. The MOEs for acrylamide are clearly below 10,000, but the Scientific Committee of EFSA gives no guidance for risk managers how to interpret a MOE less than 10,000. However, if a MOE lower than 10,000 had no meaning, the entire exercise would be worthless. A reasonable conclusion is that the MOE for acrylamide gives rise for concern and merits risk management action. Although open for criticism, using the life time risk estimates based on extrapolation and inclusion of the scaling factor the average and high exposure estimates is associated with life time risks of 1.4 to 6.4•10-3, a risk that would be considered a public health concern. The theoretical risk estimates given above are compatible with the non-positive outcome for cancer risk of acrylamide in the epidemiological studies based on FFQ. The cancer risk associated with food borne acrylamide exposure is probably low, and can at most only explain a small fraction of cancers associated with diet. However, for other chemicals a risk level to consumers comparable with that of acrylamide would not be ignored. Actions to reduce the risk by reducing acrylamide in foods and thereby food borne exposure are therefore warranted and generally agreed upon. Lastly, as it is known that a great number of proven or suspected carcinogens, e.g. other Maillard reaction products, such as heterocyclic amines, furfural and furan derivatives together with lipid peroxidation products can be formed during heat treatment of food, it is important to further study the health risk associated with heat-generated food toxicants foods. Acrylamide is not the sole problem.

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Acrylamide is not the sole problem.

To summarize, the main conclusions are: • Acrylamide has been classified by WHO, World Health organisation. This conclusion is

strengthened by the project. Following exposure to acrylamide via food the primary concern is the possible risk of cancer.

• Compared with many regulated food carcinogens, the exposure of acrylamide poses a higher estimated risk to European consumers.

• Risk assessments and recommendations to minimize exposure to acrylamide made by WHO are still valid.

• Other compounds formed during cooking of food, for example HMF, Furan, and a variety of Maillard reactants and lipid oxidation products may also constitute an increased cancer risk for consumers. Approximately 50 substances that would require risk assessment have been theoretically identified within the project.

• Current knowledge does not allow for a risk-benefit assessment of cooking with respect to acrylamide or other heat-induced toxicants.

Acrylamide Furan HMF

52 “suspect” compounds

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6.2. Risk communication Risk communication strategy and actions have been integrated with research activities in the project. Different kinds of strategic communication tools and channels have been applied.

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Figure 23. The HEATOX ID was created early in the project to illustrate the main strategic grips and to help researchers focus on the communication tasks.

6.2.1. Risk Communication Strategy To support and strengthen the project profile a Risk Communication Strategy was developed. The strategy was based on the overall HEATOX objectives and an initial analysis of communication challenges based on interviews with key persons within the project and representatives from interested parties. The strategy framework was set up in 2004 and offered as a guideline for risk communication within the project and between the project and the outside world. Main channels and a visual profile for the project was outlined and developed early in the project. Based on own observations and comments in Midterm Review Report, efforts were taken to strengthen risk communication and dissemination activities within HEATOX. During summer 2005, a dialogue was initiated with HEATOX External Panel. The aim being to ensure that end deliverables were as useful and focused as possible and that dissemination from HEATOX to the outside world was as effective as possible. Special risk communication training sessions have been organised targeting the PhDs and young researchers in the project. Considerable attention was given to the designing and organising of the main risk communication event of HEATOX, the Workshop in June 2006. The conclusions from the HEATOX workshop have given clear indications as to how some of the end deliverables should be constructed to meet end user’s needs and expectations.

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Did risk communication play the intended role in the HEATOX project? What lessons can be learned? Can HEATOX experiences contribute to making Guidelines for good risk communication practice related to heat-induced toxicants? At the end of the project, evaluations have been organised to try to answer these questions and give further insight into the nature of practical risk communication. To assess the risk communication efforts within HEATOX, two limited web-based surveys were conducted during January 2007. To assess the risk communication efforts between HEATOX and the outside world, short personal interviews with members of the External Panel were conducted in February 2007. The interviews focused on possible beneficiaries from HEATOX work, to what extent HEATOX has succeeded in networking with the outside world. The Panel members were also asked to give their personal reflections related to being members of the Panel. 6.2.1.1. HEATOX Advice to Industry and Catering Industry and catering represent important food chain levels where effective risk minimisation initiatives could contribute to reducing the intake of acrylamide. HEATOX research covers many aspects related to the three main food sectors; • Potato products; from growing conditions to crisp or French fry processing • Cereal products; mainly focused on bread and baking • Coffee; roasting process and influence of raw material parameters. There has been active and close contact between CIAA (the Confederation of the Food and Drink Industries of the EU) and HEATOX during the entire project period. The first version of the CIAA Acrylamide Toolbox was evaluated internally within HEATOX by relevant research groups prior to the decision to build further on this basis rather than developing an alternative HEATOX system. Following advice from the HEATOX External Panel and the outcome of the HEATOX Workshop in 2006, HEATOX results relevant to industry and catering sector have been fed into the CIAA Toolbox. 6.2.1.2. HEATOX Advice to Authorities and Consumer Organisations on Home Cooking and Consumption Acrylamide formation and minimisation strategies have been quite extensively studied in the laboratory and in industrial environments where the processing/heating conditions are better controlled than in home cooking. HEATOX researchers have conducted a number of experiments related to deep-fried French fries, crisps, oven-roasted potato wedges and toasted bread. The general intake of acrylamide for adults is quite similar across Europe. Due to the fact that a large number of foods contributing to acrylamide intake are industrially produced the contribution from home cooking is probably quite small in the general population. The HEATOX Workshop in Graz 2006 identified significant differences in home cooking methods and availability of ingredients within countries and certainly across the different European regions and member states. The national differences in dietary habits and cooking practices, as well as the different availability of ingredients, need to be taken into account by national authorities and consumer organisations when developing material like brochures, web pages, and presentations for consumers. The HEATOX advice is aimed as a tool for National Authorities and Consumer organisations for handling the acrylamide issue in relation to home-cooking. This might include giving cooking advice directly to consumers as well as influencing providers of raw materials, pre-fried products and frying equipment for domestic use.

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Home cooking National authorities should highlight the following: Potatoes low in sugar • Low sugar potato varieties • Maintenance of suitable storage temperature during the supply chain • Low sugar levels in prefabricated potato products for domestic frying Best frying temperature • Frying temperature in the range 145 to 170°C for deep frying potatoes • Clear and accurate cooking instruction on the package of pre-fried products • Clear and accurate instruction for fryers for domestic use Golden, not brown! • French fries and roast potatoes cooked to a golden-yellow rather than golden-brown colour • Bread toasted to the lightest colour acceptable Consumption Balance the diet as proposed in national dietary recommendations and integrate acrylamide considerations into the “normal” dietary recommendations. 6.2.1.3. Guidelines for Good Risk Communication Practice related to heat-induced toxicants Objectives related to integrating risk communication activities into HEATOX form the basis for one deliverable: Guidelines to Good Risk Communication Practice related to heat-induced toxicants. Risk communication within HEATOX has had positive effects on the outcome of the project. On a short term basis, the results from the project will be more clearly disseminated towards the targeted audiences. On a long term basis, the HEATOX experience has probably motivated most of the younger researchers and many of the senior researchers to engage in risk communication activities and thus bridging the gap between science and the end users of scientific results. The multidisciplinary and holistic approach has been recognised as important and this will probably influence the final project results. Both senior scientists in capacity of theme and workpackage leaders and the PhDs have taken actively part in risk communication within HEATOX. The planned actions taken towards different stakeholders throughout the project at specific occasions have probably influenced the outcome in a positive way. Many HEATOX scientists have taken actively part in numerous risk communication activities directed towards other stakeholders. The HEATOX network, with the External Panel and Scientific Committee as core has probably had some, but limited, effects on the outcome of the project. Several of the members have been engaged in the scientific dialogue at the HEATOX meetings. The Panel played an important role in recruiting stakeholders to the HEATOX workshop and several of them also played important roles during the actual workshop. The External Panel has contributed to bringing outside knowledge and experiences to the HEATOX table and to some extent also disseminated results on behalf of HEATOX. Some have definitely functioned as HEATOX ambassadors while others have played a less significant role in the HEATOX network. The Guidelines to Good Risk Communication Practice related to heat-induced toxicants have been constructed on the basis of experiences from earlier and ongoing risk communication projects and practices, updated risk communication research and experiences from the HEATOX project. If communicated properly, it is anticipated that these guidelines could have crossover effect on other similar project, enhancing the integration of risk communication into risk-related research even further.

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Acknowledgement

The HEATOX project has been carried out as a wide international collaboration with integration of different and complementary research fields, creating a big, interesting and useful network. The success of the HEATOX project has not only been the completion of the tasks and deliverables, but the wide dissemination of results for the benefit of industry, authority and consumers. Dissemination has been achieved (and will continue) as scientific papers for refereed journals; totally around 60 papers so far, or by attendance at national and international meetings and workshops with posters and paper presentations; papers in other journals and presentations in radio and TV. Totally around 20 PhD students or young researchers have been involved in the project, and contributed with enthusiasm, curiosity and thus helped in creating a warm and stimulating atmosphere. Some have already got their Doctoral degree and several Thesis will be presented during the coming years, and I am very proud of the thought that the HEATOX project has been a part of this research. I wish to express my sincere gratitude to all HEATOX partners and members of the External panel for all work and discussions, and to our scientific officers, Achim Boenke who was in charge at the start of the HEATOX project, and Jürgen Lucas who has been our guide throughout most of the project time. It has been a pleasure to work with you and I am looking forward to seeing you again. Lund, April 2007 Life’s truest happiness lies in the friendship we make on the way…

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proc

essi

ng

cont

amin

ants

Publ

icat

ion

Food

Ind

ustr

ies

Food

Age

ncie

s 20

07-

2008

P12

Tab

ulat

ed in

form

atio

n.

Kno

wle

dge

of

met

abol

ism

and

in

tern

al d

oses

of

acry

lam

ide

(and

its

met

abol

ite

glyc

idam

ide)

Impr

oved

ca

ncer

ris

k as

sess

men

t fo

r ac

ryla

mid

e

1. A

utho

riti

es

(exp

osur

e lim

its,

e.

g. A

DI)

2.

Can

cer

rese

arch

(e

pide

mio

logy

)

P14

A

mai

n co

mpo

nent

in a

n im

prov

ed c

ance

r ri

sk

asse

ssm

ent o

f acr

ylam

ide

is c

ompa

riso

n of

inte

rnal

do

ses

of g

lyci

dam

ide

- th

e ge

noto

xic

met

abol

ite

of

acry

lam

ide

- in

hum

ans

and

anim

als

that

hav

e be

en

used

in c

ance

r te

sts.

Oth

er p

aram

eter

s th

at a

re

nece

ssar

y (i

nter

med

iate

goa

ls)

for

esti

mat

ion

of th

e ra

nge

of r

isk:

inte

r- a

nd in

tra-

indi

vidu

al v

aria

tion

in

inte

rnal

dos

es, b

ioav

aila

bilit

y of

acr

ylam

ide

in

diff

eren

t foo

dstu

ffs,

pos

sibl

e en

doge

nous

form

atio

n of

ac

ryla

mid

e.

Prob

abili

stic

ex

posu

re c

alcu

lati

on

Mon

te C

arlo

R

isk

Ass

essm

ent

prog

ram

me

(MC

RA

) fo

r sc

enar

io

anal

ysis

of

the

effe

ct o

n ex

posu

re o

f ac

ryla

mid

e re

duct

ion.

1. P

olic

y 2.

Ind

ustr

y 20

07

2008

- -

P17

M

CR

A h

as b

een

deve

lope

d in

oth

er p

roje

cts

and

is in

th

e co

nsor

tium

agr

eem

ent

defin

ed a

s pr

e-ex

isti

ng

know

ledg

e. T

he u

se o

f MC

RA

in a

cryl

amid

e an

d ot

her

heat

-tre

ated

toxi

cant

s ha

s be

en e

xplo

ited

and

w

ill b

e fu

rthe

r ex

ploi

ted

for

rese

arch

que

stio

ns o

f Fo

od I

ndus

try

(in

coop

erat

ion

wit

h IL

SI)

rega

rdin

g th

e ef

fect

iven

ess

of s

ever

al m

itig

atio

n st

eps

as

prop

osed

in th

e T

OO

LBO

X o

f the

CIA

A. F

urth

er

appl

icat

ion

of th

e M

CR

A s

oftw

are

also

in r

elat

ion

to

quan

tita

tive

ris

k-be

nefit

eva

luat

ions

are

fore

seen

. T

here

is a

lso

an in

tere

st o

f nat

iona

l Foo

d St

anda

rd

Age

ncie

s to

use

the

MC

RA

sof

twar

e in

coo

pera

tion

w

ith

the

RIK

ILT

in th

e ne

arby

futu

re.

of

77

Exp

loit

able

K

now

ledg

e (d

escr

ipti

on)

Exp

loit

able

pr

oduc

t(s)

or

m

easu

re(s

)

Sect

or(s

) of

ap

plic

atio

n

Tim

etab

le

for

com

mer

cial

us

e

Pat

ents

or

othe

r IP

R

prot

ecti

on

Ow

ner

&

Oth

er

Par

tner

(s)

invo

lved

Com

men

ts

Iden

tific

atio

n of

SU

LT1A

1 as

the

prin

cipa

l hum

an

enzy

me

invo

lved

in

the

toxi

ficat

ion

of

HM

F

Mou

se li

nes

hum

aniz

ed

for

SULT

1A1

1.

Phar

mac

euti

cal

Indu

stry

2.

Tox

icol

ogic

al

cont

ract

rse

arch

2008

or

2009

A

pplic

atio

n pa

tent

en

visa

ged

for

2008

P18

Tox

icok

inet

ics

rela

ted

to

acry

lam

ide

or

glyc

idam

ide

P22

Exp

ress

ion

prof

iling

of h

uman

MC

F7 c

ells

exp

osed

to

diff

eren

t con

cent

rati

ons

of a

cryl

amid

e or

gly

cida

mid

e R

eal-

tim

e re

vers

e-tr

ansc

ript

ase

PCR

val

idat

ion

of 2

5 tr

ansc

ript

s in

MC

F7 c

ells

and

hum

an C

aCo-

2 ce

lls

Mea

sure

men

t of

intr

acel

lula

r gl

utat

hion

e le

vels

aft

er

trea

tmen

t wit

h ac

ryla

mid

e or

gly

cida

mid

e D

eter

min

atio

n of

nuc

lear

fact

or-•

B a

ctiv

ity

usin

g a

repo

rter

gen

e as

say

Stan

dard

toxi

city

and

cel

l via

bilit

y as

says

.

Kno

wle

dge

on

vacu

um fr

ying

to

redu

ce a

cryl

amid

e le

vel i

n po

tato

es

cris

ps

In

dust

ry

P23

Vac

uum

fryi

ng w

ith

and

wit

hout

pre

-tre

atm

ents

has

sh

own

to p

rodu

ce lo

w le

vel a

cryl

amid

e cr

isps

. It i

s po

ssib

le to

impr

ove

thei

r qu

alit

y ch

arac

teri

stic

s. T

he

pre-

trea

tmen

ts r

epre

sent

a p

ract

ical

indu

stri

al

alte

rnat

ive

to r

educ

e ac

ryla

mid

e le

vel i

n po

tato

es

cris

ps

of

77

Dis

sem

inat

ion

of K

now

ledg

e SC

IEN

TIF

IC J

OU

RN

AL

Yea

r 20

04

Par

tner

A

utho

r

Jour

nal

Tit

le

See

list,

pa

ge 6

. M

urko

vic,

M.

J. B

ioch

em. B

ioph

ys. M

eth.

61,

161

-16

7, 2

004

Acr

ylam

ide

in A

ustr

ian

Food

s P2

Surd

yk, N

., R

osen

, J.,

And

erss

on, R

. and

Åm

an, P

J.

Agr

ic. F

ood

Che

m.

Eff

ects

of a

spar

agin

es, f

ruct

ose,

and

bak

ing

cond

itio

ns o

n ac

ryla

mid

e co

nten

t in

yeas

t-le

aven

ed w

heat

bre

ad

P4

Fred

riks

son,

H.,

Tal

lvin

g, J

, R

osen

, J. a

nd Å

man

, P.

Cer

eal C

hem

. Fe

rmen

tati

on r

educ

es fr

ee a

spar

agin

es in

dou

gh a

nd a

cryl

amid

e co

nten

t in

brea

d P4

Hus

øy, T

., A

bram

sson

-Z

ette

rber

g, L

., Ø

lstø

rn, H

., Pa

ulse

n J

E.,

and

Ale

xand

er J

.

Mut

atio

n R

esea

rch,

580

, 103

-110

. A

deno

mat

ous

poly

posi

s co

li in

fluen

ces

the

effe

ct o

f the

hea

t ind

uced

m

utag

ens,

PhI

P an

d ac

ryla

mid

e, o

n th

e pr

oduc

tion

of m

icro

nucl

ei in

m

ice

eryt

hroc

ytes

.

P9

Dur

ling,

L. a

nd L

ilian

ne

Abr

amss

on-Z

ette

rber

g, L

.*

Mut

atio

n R

esea

rch,

580

, 103

-110

.

A c

ompa

riso

n of

gen

otox

icit

y be

twee

n th

ree

com

mon

het

eroc

yclic

am

ines

and

acr

ylam

ide.

P9

Dun

ovsk

á L.

, Haj

šlov

á J.

, aj

ka

T.,

Hol

adov

á K

. and

Háj

ková

K.

Cze

ch J

ourn

al o

f Foo

d Sc

ienc

e, V

ol.

22, 2

83-2

86

Cha

nges

of A

cryl

amid

e Le

vels

in F

ood

Prod

ucts

dur

ing

Tec

hnol

ogic

al

Proc

essi

ng

P10

E. B

erm

udo,

V. R

uiz-

Cal

ero,

L.

Puig

nou,

M.T

. Gal

cera

n E

lect

roph

ores

is

Mic

roem

ulsi

on

elec

trok

inet

ic

chro

mat

ogra

phy

for

the

anal

ysis

of

ac

ryla

mid

e in

food

. P1

2

Year

200

5

Hel

lenä

s K

.-E

., A

bram

sson

-Z

ette

rber

g L.

, and

Sko

g K

. Jo

urna

l of A

OA

C I

nter

nati

onal

, Ja

nuar

y/Fe

brua

ry V

ol. 8

8, N

o. 1

, 242

-24

5.

The

HE

AT

OX

pro

ject

,

P1

P9

Bag

dona

ite,

K.,

Mur

kovi

c, M

. C

zech

J. F

ood

Sci.

22, 2

2-24

, 200

5 Fa

ctor

s af

fect

ing

the

form

atio

n of

acr

ylam

ide

in c

offe

e P2

Mus

tafa

, A.,

Ade

rsso

n, R

., R

osen

, J.,

Kam

al-E

ldin

A. a

nd

Åm

an P

.

J. A

gric

. Foo

d C

hem

. 200

5, 5

3, 5

985-

5989

Fa

ctor

s in

fluen

cing

acr

ylam

ide

cont

ent a

nd c

olor

in r

ye c

risp

bre

ad

P4

Ols

son,

E.E

.M.,

Trä

gård

h, A

.C.

& A

hrné

, L.M

. J.

Foo

d Sc

i. 70

(8),

E48

4-49

1 (2

005)

E

ffec

t of N

ear-

infr

ared

Rad

iati

on a

nd J

et I

mpi

ngem

ent H

eat T

rans

fer

on C

rust

For

mat

ion

of B

read

P6

of

77

Kno

l,J.J

; Van

Loo

n,W

.A.M

. van

; Li

nsse

n,J.

P.H

.; R

uck,

A.L

.; V

an

Boe

kel,M

.A.J

.S.;

Vor

agen

,A.G

J.

J. o

f Agr

ic. F

ood

Che

m. 2

005,

53,

61

33-6

139.

T

owar

d a

Kin

etic

Mod

el fo

r A

cryl

amid

e Fo

rmat

ion

in a

Glu

cose

-A

spar

agin

e R

eact

ion

Syst

em.

P7

L. C

astle

and

S. E

riks

son

Jour

nal o

f AO

AC

Int

erna

tiona

l, 20

05,

88, 2

74-2

84.

Ana

lyti

cal m

etho

ds u

sed

to m

easu

re a

cryl

amid

e co

ncen

trat

ions

in

food

s.

P8

L. M

. Ow

en, L

. Cas

tle, J

. Kel

ly,

L. W

ilson

and

A. S

. Llo

yd

Jour

nal o

f AO

AC

Int

erna

tiona

l, 20

05,

88, 2

85-2

91.

Acr

ylam

ide

anal

ysis

: Ass

essm

ent o

f res

ults

from

six

rou

nds

of F

ood

Ana

lysi

s Pe

rfor

man

ce A

sses

smen

t Sch

eme

(FA

PAS)

pro

ficie

ncy

test

ing.

P8

P. F

ohge

lber

g, J

. Ros

én,

K.-

E.

Hel

lenä

s, L

. Abr

amss

on-

Zet

terb

erg*

Food

and

Che

mic

al T

oxic

olog

y, 4

3: 9

51-

959.

T

he a

cryl

amid

e in

take

via

som

e co

mm

on b

aby

food

for

child

ren

in

Swed

en d

urin

g th

eir

first

yea

r of

life

- (

an im

prov

ed m

etho

d fo

r an

alys

is o

f acr

ylam

ide)

.

P9

Pete

rsso

n E

.V.,

Ros

én J

., T

urne

r C

., D

anie

lsso

n R

., H

elle

näs

KE

. A

naly

tica

Chi

mic

a A

cta

Cri

tica

l fac

tors

and

pit

falls

aff

ecti

ng th

e ex

trac

tion

of a

cryl

amid

e fr

om

food

s: A

n op

tim

isat

ion

stud

y

P9

Dun

ovsk

á L.

, aj

ka T

. and

H

ajšl

ová

J.

Che

mic

ké li

sty

Jour

nal,

99 2

79-2

80,

2005

, Spe

cial

Iss

ue 1

4 A

cryl

amid

e Le

vels

in F

oods

tuff

s fr

om C

zech

mar

ket

P10

E. B

erm

udo,

O. N

úñez

, L.

Puig

nou,

M.T

. Gal

cera

n Jo

urna

l of C

hrom

atog

raph

y A

A

naly

sis

of a

cryl

amid

e in

food

sam

ples

by

capi

llary

zon

e el

ectr

opho

resi

sP1

2

E. B

arce

ló-B

arra

chin

a, F

.J.

Sant

os, L

. Pui

gnou

, M.T

. G

alce

ran

Ana

lyti

ca C

him

ica

Act

a D

eter

min

atio

n of

het

eroc

yclic

am

ines

in a

mea

t ext

ract

as

dim

ethy

lform

amid

e di

alky

lace

tal d

eriv

ativ

es b

y ga

s ch

rom

atog

raph

y m

ass

spec

trom

etry

.

P12

Hag

mar

, L.,

Wir

fält

E.,

Paul

sson

, B.,

Tör

nqvi

st, M

. M

utat

ion

Res

earc

h 58

0, 1

57.1

65

Diff

eren

ces

in h

emog

lobi

n ad

duct

leve

ls o

f acr

ylam

ide

in th

e ge

nera

l po

pula

tion

wit

h re

spec

t to

diet

ary

inta

ke, s

mok

ing

habi

ts a

nd g

ende

r P1

4

Tho

mas

Bje

llaas

, Kar

el J

anak

, E

lsa

Lund

anes

, Geo

rg B

eche

r

Xen

obio

tica

D

eter

min

atio

n an

d qu

anti

ficat

ion

of u

rina

ry m

etab

olit

es a

fter

die

tary

ex

posu

re to

acr

ylam

ide

P16

Boo

n PE

, Mul

A d

e, V

oet H

van

de

r, D

onke

rsgo

ed G

van

, Bre

tte

M, K

lave

ren

JD v

an

Mut

atio

n R

esea

rch

580,

143

-155

; 20

05

Cal

cula

tion

s of

die

tary

exp

osur

e to

acr

ylam

ide

P17

H. R

. Gla

tt, H

. Sch

neid

er, Y

.-G

. Li

u

Mut

atio

n R

esea

rch

580

(200

5) 4

1-52

V

79-h

CY

P2E

1-hS

ULT

1A1,

a c

ell l

ine

for

the

sens

itiv

e de

tect

ion

of

geno

toxi

c ef

fect

s in

duce

d by

car

bohy

drat

e py

roly

sis

prod

ucts

and

oth

er

food

-bor

ne c

hem

ical

s

P18

H. R

. Gla

tt, W

. Mei

nl

Met

hods

in E

nzym

olog

y 40

0 (2

005)

: 23

0-24

9 Su

lfotr

ansf

eras

es a

nd a

cety

ltran

sfer

ases

in m

utag

enic

ity

test

ing:

te

chni

cal a

spec

ts

P18

of

77

Yea

r 20

06

Bag

dona

ite,

K.,

Vik

lund

, G.,

Skog

, K.,

Mur

kovi

c, M

. J.

Bio

chem

. Bio

phys

. Met

hods

, 69,

21

5-22

1, 2

006

Ana

lysi

s of

3-a

min

opro

pion

amid

e: A

pot

enti

al p

recu

rsor

of a

cryl

amid

e P2

, P1

Mur

kovi

c, M

., Pi

chle

r, N

. M

olec

ular

Nut

riti

on &

Foo

d R

esea

rch,

50

, 842

-846

(20

06)

Ana

lysi

s of

5-h

ydro

xym

ethy

lfurf

ural

in c

offe

e, d

ried

frui

ts a

nd u

rine

. P2

Mur

kovi

c, M

., D

erle

r, K

. J.

Bio

chem

. Bio

phys

. Met

hods

, 69,

25-

32, 2

006

Ana

lysi

s of

am

ino

acid

s an

d ca

rboh

ydra

tes

in g

reen

cof

fee.

P2

Low

, M. Y

.; K

outs

idis

, G.;

Park

er, J

. K.;

Elm

ore,

J. S

.; D

odso

n, A

. T.;

Mot

tram

, D. S

.

Jour

nal o

f Agr

icul

tura

l and

Foo

d C

hem

istr

y E

ffec

t of c

itri

c ac

id a

nd g

lyci

ne a

ddit

ion

on a

cryl

amid

e an

d fla

vor

in a

po

tato

mod

el s

yste

m.

NB

Wor

k pa

rtly

supp

orte

d by

Hea

tox

P3

Mar

tin

Fink

, Rog

er A

nder

sson

, Jo

han

Ros

én, a

nd P

er Å

man

C

erea

l Che

mis

try

Eff

ect o

f Add

ed A

spar

agin

e an

d G

lyci

ne o

n A

cryl

amid

e C

onte

nt in

Y

east

-Lea

vene

d B

read

P4

S. R

oman

i*, A

. Gas

parr

i*, M

. B

acch

iocc

a*, E

. Coc

ci*,

P. R

occu

li*,

M. D

alla

Ros

a*

Indu

stri

e al

imen

tari

(c

hiri

otti

ed.

, pin

erol

o –

to –

ital

y)

Con

tenu

to in

acr

ilam

mid

e e

cara

tter

isti

che

qual

itat

ive

di p

atat

e fr

itte

in

diff

eren

ti a

ppar

ati t

ecno

logi

ci –

Acr

ylam

ide

cont

ent a

nd c

hara

cter

siti

cs

of p

otat

o fr

ied

in d

iffer

ent f

ryer

s

P5

S. R

oman

i, M

. Bac

chio

cca,

P.

Roc

culi,

M. D

alla

Ros

a A

ccep

ted

to b

e pu

blis

hed

on E

urop

ean

Food

Res

earc

h A

nd T

echn

olog

y E

ffec

t of f

ryin

g ti

me

on a

cryl

amid

e co

nten

t and

qua

lity

aspe

cts

of

Fren

ch fr

ies

P5

Wen

zl, T

., K

aras

ek, L

., R

osen

, J.

, Hel

lena

es, K

-E.,

Cre

ws,

C.,

Cas

tle, L

. and

Ank

lam

, E.

Jour

nal

of

Chr

omat

ogra

phy

A.

2006

11

32,

211-

21.

Col

labo

rati

ve tr

ial v

alid

atio

n st

udy

of tw

o m

etho

ds, o

ne b

ased

on

high

pe

rfor

man

ce li

quid

chr

omat

ogra

phy–

tand

em m

ass

spec

trom

etry

and

on

gas

chr

omat

ogra

phy–

mas

s sp

ectr

omet

ry fo

r th

e de

term

inat

ion

of

acry

lam

ide

in b

aker

y an

d po

tato

pro

duct

s.

P8

Has

nip,

S.,

Cre

ws,

C. a

nd

Cas

tle, L

. Fo

od A

ddit

ives

and

Con

tam

inan

ts,

2006

, 23

, 219

-227

. So

me

fact

ors

affe

ctin

g th

e fo

rmat

ion

of fu

ran

in h

eate

d fo

ods.

P8

Gol

man

n, T

.,Per

isse

t, A

., B

erth

olet

, M.-

C.,

Stad

ler,

R. H

., Pe

ters

son,

E. V

., H

elle

näs,

K.-

E.

Food

Add

it. C

onta

m.,

23(5

), 4

37-4

45

Impa

ct o

f ext

ract

ion

cond

itio

ns o

n th

e co

nten

t of a

cryl

amid

e in

mod

el

syst

ems

and

food

. P9

Wen

zl, T

., K

aras

ek, L

., R

osén

, J.

, Hel

lena

es, K

.-E

., C

rew

s, C

., C

astle

, L.,

and

Ank

lam

, E.

J. C

hrom

. A.,

doi:1

0.10

16/j

.chr

oma.

2006

.07.

007

Col

labo

rati

ve tr

ial v

alid

atio

n st

udy

of tw

o m

etho

ds, o

ne b

ased

on

high

pe

rfor

man

ce li

quid

chr

omat

ogra

phy–

tand

em m

ass

spec

trom

etry

and

on

gas

chr

omat

ogra

phy–

mas

s sp

ectr

omet

ry fo

r th

e de

term

inat

ion

of

acry

lam

ide

in b

aker

y an

d po

tato

pro

duct

s.

P9

Dun

ovsk

á L.

, aj

ka T

., H

ajšl

ová

J., H

olad

ová

K.

Ana

lyti

ca C

him

ica

Act

a, V

olum

e 57

8,

Issu

e 2,

25

Sept

embe

r 20

06, 2

34-2

40

Dir

ect d

eter

min

atio

n of

acr

ylam

ide

in fo

od b

y ga

s ch

rom

atog

raph

y-hi

gh-r

esol

utio

n ti

me-

of-f

light

mas

s sp

ectr

omet

ry

P10

of

77

E. B

erm

udo,

O. N

úñez

, L.

Puig

nou,

M.T

. Gal

cera

n J.

Chr

omat

ogr.

A, 1

120

(200

6) 1

99-

204

A

naly

sis

of a

cryl

amid

e in

food

sam

ples

by

capi

llary

zon

e el

ectr

opho

resi

sP1

2

E. B

erm

udo,

O. N

úñez

, L.

Puig

nou,

M.T

. Gal

cera

n J.

Chr

omat

ogr.

A, 1

129

(200

6) 1

29-

134

Ana

lysi

s of

acr

ylam

ide

in fo

od p

rodu

cts

by in

-lin

e pr

econ

cent

rati

on

capi

llary

zon

e el

ectr

opho

resi

s P1

2

E. T

eixi

dó, F

.J. S

anto

s, L

. Pu

igno

u, M

.T. G

alce

ran

J. C

hrom

atog

r. A

, 113

5 (2

006)

85-

90

Ana

lysi

s of

5-h

ydro

xym

ethy

lfurf

ural

in fo

ods

by g

as c

hrom

atog

raph

y-m

ass

spec

trom

etry

P1

2

T B

jella

as, L

H S

tøle

n, M

H

auge

n, J

E P

auls

en, J

A

lexa

nder

, E L

unda

nes

and

G

Bec

her

Food

and

Che

mic

al T

oxic

olog

y

2006

Dec

19;

[E

pub

ahea

d of

pri

nt]

Uri

nary

acr

ylam

ide

met

abol

ites

as

biom

arke

rs fo

r sh

ort-

term

die

tary

ex

posu

re to

acr

ylam

ide.

P1

6

Year

200

7

G. V

iklu

nd, F

. Men

doza

, I.

Sjö

holm

and

K. S

kog

LW

T -

Foo

d Sc

ienc

e an

d T

echn

olog

y,

40, i

ssue

6, 1

066-

1071

A

n ex

peri

men

tal s

et-u

p fo

r st

udyi

ng a

cryl

amid

e fo

rmat

ion

in p

otat

o cr

isps

P1

G. V

iklu

nd,.

Ols

son,

K.,

Sjöh

olm

I. a

nd K

. Sko

g su

bmit

ted

Eff

ect o

f pot

ato

vari

ety,

yea

r an

d st

orag

e co

ndit

ions

for

the

form

atio

n of

acr

ylam

ide

in c

risp

s P1

G. V

iklu

nd,.

Ols

son,

K.,

Sjöh

olm

I. a

nd K

. Sko

g su

bmit

ted

Bla

nchi

ng r

educ

es p

recu

rsor

con

tent

and

acr

ylam

ide

form

atio

n in

fo

rmat

ion

in c

risp

s P1

Mus

tafa

, A.,

Åm

an, P

., A

nder

sson

, R. K

amal

-Eld

in A

. Fo

od C

hem

istr

y,

subm

itte

d A

naly

sis

of fr

ee a

min

o ac

ids

in c

erea

l pro

duct

s P4

Ahr

ne, L

., A

nder

sson

, C.-

G.,

Flob

erg,

P.,

Ros

én, J

. &

Ling

nert

, H.

LWT

– F

ood

Scie

nce

and

Tec

hnol

ogy,

In

pre

ss

Eff

ect o

f cru

st te

mpe

ratu

re a

nd w

ater

con

tent

on

acry

lam

ide

form

atio

n du

ring

bak

ing

of w

hite

bre

ad: s

team

and

falli

ng te

mpe

ratu

re b

akin

g P6

J.J.

Kno

l, G

. Vik

lund

, J.P

.H.

Lins

sen,

I. S

jöho

lm, K

. Sko

g,

M.A

.J.S

. van

Boe

kel

Jour

nal o

f Mol

ecul

ar N

utri

tion

and

Fo

od R

esea

rch

Kin

etic

Mod

elin

g: A

Too

l to

Und

erst

and

the

Form

atio

n of

Acr

ylam

ide

in F

ood

In p

repa

ratio

n

P7, P

1

C. C

rew

s an

d L.

Cas

tle

Tre

nds

in F

ood

Scie

nce

and

Tec

hnol

ogy,

in p

ress

R

evie

w o

f the

occ

urre

nce,

form

atio

n an

d an

alys

is o

f fur

an in

hea

t-pr

oces

sed

food

s P8

C. C

rew

s, S

Has

nip,

D. P

. T.

Rob

erts

and

L. C

astle

Fo

od A

ddit

ives

& C

onta

min

ants

,

in p

ress

. Fa

ctor

s af

fect

ing

the

anal

ysis

of f

uran

in h

eate

d fo

ods.

P8

C. C

rew

s an

d L.

Cas

tle.

LC-G

C E

urop

e, in

pre

ss

The

det

erm

inat

ion

of fu

ran

in fo

ods-

cha

lleng

es a

nd s

olut

ions

. P8

D. P

. T. R

ober

ts, C

. Cre

ws

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H. G

rund

y M

anus

crip

t in

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arat

ion.

E

ffec

t of c

onsu

mer

coo

king

on

fura

n in

con

veni

ence

food

s

P8

of

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Vol

lebr

egt,

H.M

.; Z

onde

rvan

, C

. J.

Hea

t Tra

nsfe

r A

kin

etic

and

ther

mod

ynam

ic m

odel

to p

redi

ct le

vels

of a

cryl

amid

e in

he

ated

food

pro

duct

s (t

enta

tive

) P1

1

M.S

. Alta

ki, F

.J. S

anto

s, M

.T.

Gal

cera

n J.

Chr

omat

ogr.

A

Ana

lysi

s of

fura

n in

food

s by

hea

dspa

ce s

olid

-pha

se m

icro

extr

acti

on-g

as

chro

mat

ogra

phy-

ion

trap

mas

s sp

ectr

omet

ry

P12

E. B

erm

udo,

O. N

úñez

, E.

Moy

ano,

L. P

uign

ou, M

.T.

Gal

cera

n

J. C

hrom

atog

r. A

Fi

eld

Am

plifi

ed S

impl

e In

ject

ion-

Cap

illar

y E

lect

roph

ores

is-T

ande

m

Mas

s Sp

ectr

omet

ry fo

r th

e an

ális

is o

f acr

ylam

ide

in fo

odst

uffs

P12

A V

ikst

röm

, S E

riks

son,

B

Paul

sson

, P K

arls

son

and

M

Tör

nqvi

st

subm

itte

d C

ompa

riso

n of

bio

avai

labi

lity

of a

cryl

amid

e in

diff

eren

t foo

ds-a

stu

dy

in m

ice

P14

E W

irfä

lt, B

Pau

lsso

n, M

T

örnq

vist

, A A

xmon

and

L

Hag

mar

Eur

opea

n Jo

urna

l of C

linic

al N

utri

tion

(i

n pr

ess,

doi

10.

1038

/sj.e

jcn.

1602

704)

Ass

ocia

tion

s be

twee

n es

tim

ated

acr

ylam

ide

(AA

) in

take

s, a

nd H

b A

A-

addu

cts

in a

sam

ple

from

the

Mal

mö

Die

t and

Can

cer

coho

rt

P14

T B

jella

as*,

† , P T

honn

ing

Ole

sen‡ , H

Fra

ndse

n‡ , M

Hau

gen* , L

L-H

Stø

len* , J

E

Paul

sen* , J

Ale

xand

er* , E

Lu

ndan

es† a

nd G

Bec

her*,

†

Tox

icol

ogic

al S

cien

ces,

sub

mit

ted

Com

pari

son

of e

stim

ated

die

tary

inta

ke o

f acr

ylam

ide

wit

h ha

emog

lobi

n ad

duct

s of

acr

ylam

ide

and

glyc

idam

ide

P15

R.V

. Hed

egaa

rd, H

. Fra

ndse

n,

K. G

ranb

y, A

. Apo

stol

opou

la

and

L. S

kibs

ted

Jour

nal o

f foo

d an

d A

gric

ultu

ral

Che

mis

try,

sub

mit

ted

Influ

ence

of w

ater

act

ivit

y on

acr

ylam

ide

form

atio

n fr

om g

luco

se a

nd

aspa

ragi

nes

in a

queo

us g

lyce

rol

P15

Cle

men

t F, D

ip R

, Nae

geli

H

Can

cer

Epi

dem

iolo

gy B

iom

arke

rs a

nd

Prev

enti

on (

Subm

itte

d)

Exp

ress

ion

Sign

atur

e of

Gly

cida

mid

e, th

e R

eact

ive

Met

abol

ite

of th

e W

ides

prea

d Fo

od C

arci

noge

n A

cryl

amid

e P2

2

of

77

Boo

k C

hapt

ers

Y

ear

2006

Aut

hor

Boo

k T

itle

P

artn

ner

Mot

tram

, D. S

; Low

, M. Y

.; E

lmor

e, J

. S

In: A

cryl

amid

e an

d ot

her

Haz

ardo

us

Com

poun

ds in

Hea

t-T

reat

ed F

oods

. K

Sko

g; J

Ale

xand

er (

Eds

), W

oodh

ead

Publ

ishi

ng

The

Mai

llard

rea

ctio

n an

d it

s ro

le in

the

form

atio

n of

acr

ylam

ide

and

othe

r po

tent

ially

haz

ardo

us c

ompo

unds

in fo

ods

P3

Q. C

haud

hry,

J. C

otte

rill,

R.

Wat

kins

and

N. P

rice

In

‘Acr

ylam

ide

and

othe

r he

alth

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zard

ous c

ompo

unds

in h

eat-

trea

ted

food

s’. p

p. 1

32-1

59.

K

. Sko

g an

d J.

Ale

xand

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Eds

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Woo

dhea

d Pu

blis

hing

, 200

6. I

SBN

97

8-1-

8456

9-01

1-3.

A m

olec

ular

mod

ellin

g ap

proa

ch to

pre

dict

the

toxi

city

of c

ompo

unds

ge

nera

ted

duri

ng h

eat t

reat

men

t of f

ood.

P8

L. C

astle

In

‘Acr

ylam

ide

and

othe

r he

alth

ha

zard

ous c

ompo

unds

in h

eat-

trea

ted

food

s’. p

p. 1

17-1

31.

K

. Sko

g an

d J.

Ale

xand

er (

Eds

),

Woo

dhea

d Pu

blis

hing

, 200

6. I

SBN

-13

: 978

-1-8

4569

-011

-3.

Ana

lysi

s fo

r ac

ryla

mid

e in

food

s.

P8

Oth

er P

ubli

cati

ons

Y

ear

2004

Hül

ya E

rol G

ezgi

n D

ünya

GID

A-M

ayıs

200

4 Is

ıl i

lem

Gör

en G

ıdal

arda

Kan

ser

Ris

ki (

Can

cer

Ris

k in

Hea

t Tre

ated

Fo

ods)

P13

K G

ranb

y, H

Fra

ndse

n, K

Kac

k,

BB

B J

ense

n, J

A N

iels

en

Alim

enta

A

cryl

amid

i fø

deva

rer

P15

Yea

r 20

05

Per

Åm

an

Livs

med

el I

Fok

us, 2

005,

7 p

p 57

A

kryl

amid

i br

öd

P4

J.J.

Kno

l, J.

P.H

. Lin

ssen

and

M

.A.J

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an B

oeke

l B

i-/t

rien

nial

per

iodi

cal o

n cu

rren

t re

sear

ch o

f the

gra

duat

e sc

hool

VLA

G

Kin

etic

Mod

elin

g of

Acr

ylam

ide

Form

atio

n in

Mod

el S

yste

ms

P7

of

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L C

astle

FS

A F

ood

Surv

ey I

nfor

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ion

Shee

t 71

/05.

Jan

uary

200

5.

http

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ww

.food

.gov

.uk/

mul

tim

edia

/pdf

s/fs

is71

2005

.pdf

#pag

e=5

Ana

lysi

s of

Tot

al D

iet S

tudy

sam

ples

for

acry

lam

ide.

P8

Hül

ya Ö

lmez

-Son

gün

Dem

irel

D

ünya

GID

A-O

cak

2005

T

ahıl

Baz

lı G

ıdak

lard

a A

krila

mid

(A

cryl

amid

e in

cer

eal b

ased

pr

oduc

ts)

P13

Jaco

b D

. van

Kla

vere

n, P

olly

E.

Boo

n an

d A

nika

de

Mul

W

oodh

ead

publ

ishi

ng. (

subm

itte

d)

Mod

ellin

g of

die

tary

exp

osur

e to

acr

ylam

ide,

Acr

ylam

ide

and

othe

r he

alth

haz

ardo

us c

ompo

unds

in h

eat-

trea

ted

food

s.

P17

H. R

. Gla

tt

In: H

uman

Sul

phot

rans

fera

ses (

G. M

. Pa

cific

i, M

. W. H

. Cou

ghtr

ie, e

ds.)

T

aylo

r &

Fra

ncis

, Lon

don,

p. 2

81-3

06

Act

ivat

ion

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inac

tiva

tion

of c

arci

noge

ns b

y hu

man

sul

fotr

ansf

eras

es

P18

H. R

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tt, W

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bner

, W.

Mei

nl

In: 1

5th

Inte

rnat

iona

l Sym

posiu

m o

n M

icro

som

es a

nd D

rug

Oxi

datio

ns, h

eld

in M

ainz

(G

erm

any)

(F.

Oes

ch, e

d.),

M

edim

ond/

Mon

duzz

i, B

olog

na, p

. 9-

15

Sulfo

tran

sfer

ases

exp

ress

ed in

the

hum

an g

astr

o-in

test

inal

trac

t and

th

eir

abili

ty to

act

ivat

e pr

omut

agen

s/ca

rcin

ogen

s P1

8

Yea

r 20

06

Skog

K. a

nd A

lexa

nder

J. (

Eds

) A

cryl

amid

e an

d ot

her

haza

rdou

s co

mpo

unds

in h

eat-

trea

ted

food

s,

Woo

dhea

d Pu

blis

hing

Ltd

BO

OK

wit

h co

ntri

buti

ons

of m

any

HE

AT

OX

par

tner

s P1

, P16

Skog

K, H

elle

näs

K-E

. and

Li

ngne

rt H

. Li

vsm

edel

i fo

kus

CIA

A-w

orks

hop

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g ak

ryla

mid

P1

, P6,

P9

Sk

og K

. Fo

od S

cien

ce a

nd T

echn

olog

y, in

pre

ss

The

HE

AT

OX

pro

ject

: Hea

t-ge

nera

ted

food

toxi

cant

s: I

dent

ifica

tion

, C

hara

cter

isat

ion

and

Ris

k M

inim

isat

ion,

P1

Mur

kovi

c M

Acr

ylam

ide

and

othe

r ha

zard

ous

com

poun

ds in

hea

t tre

ated

food

s, S

kog

K, A

lexa

nder

J (

Eds

.)

Woo

dhea

d Pu

blis

hing

Ltd

, Cam

brid

ge

Mec

hani

sm fo

r th

e fo

rmat

ion

of P

hIP

P2

Mur

kovi

c M

Scha

dsto

ffe

in L

eben

smit

teln

und

Fu

tter

mit

teln

, Cic

hna-

Mar

kl M

, So

ntag

G, S

tidl

R (

Eds

.) G

esel

lsch

aft

Öst

erre

ichi

sche

r C

hem

iker

, pp.

2-1

0,

Wie

n, 2

006

Bild

ung

von

hete

rozy

klis

chen

aro

mat

isch

en A

min

en b

eim

Erh

itze

n vo

n Fl

eisc

h

P2

of

77

Bag

dona

ite

K, M

urko

vic

M

Sc

hads

toff

e in

Leb

ensm

itte

ln u

nd

Futt

erm

itte

ln, C

ichn

a-M

arkl

M,

Sont

ag G

, Sti

dl R

(E

ds.)

Ges

ells

chaf

t Ö

ster

reic

hisc

her

Che

mik

er, p

p. 2

09-

214,

Wie

n, 2

006

Form

atio

n of

acr

ylam

ide

in c

offe

e

P2

Low

, M. Y

.; E

lmor

e, J

. S.;

Mot

tram

, D. S

. In

: Fla

vour

Sci

ence

- R

ecen

t Adv

ance

s an

d T

rend

s. W

. Bre

die;

M. P

eter

sen

(Eds

), E

lsev

ier.

Rel

atio

nshi

p be

twee

n ac

ryla

mid

e fo

rmat

ion

and

the

gene

rati

on o

f fla

vour

in h

eate

d fo

ods

NB

Wor

k pa

rtly

supp

orte

d by

Hea

tox

P3

Pehr

sson

, S

NyT

ekni

k, 4

8(20

06),

5

Så b

akar

du

akry

lam

idfr

itt

P6

M T

örnq

vist

, B P

auls

son

and

S O

ster

man

-Gol

kar

Acr

ylam

ide

and

othe

r ha

zard

ous

com

poun

ds in

hea

t-tr

eate

d fo

ods,

K

erst

in S

kog

and

Jan

Ale

xand

er

Bio

-mon

itor

ing

of a

cryl

amid

e P1

4

SH H

anse

n, E

J Sø

derl

und,

D

And

erso

n, A

Bau

mga

rtne

r, R

G

opal

an a

nd G

Bru

nbor

g

14th

Eur

opea

n T

esti

s W

orks

hop,

20

06. P

rogr

amm

e an

d m

inip

oste

rs

Abs

trac

t: D

NA

dam

agin

g ef

fect

s of

the

food

con

tam

inan

t acr

ylam

ide

in m

ale

germ

cel

ls

P16

Jaco

b D

. van

Kla

vere

n, P

olly

E.

Boo

n an

d A

nika

de

Mul

Boo

k: A

cryl

amid

e an

d ot

her

heal

th

haza

rdou

s co

mpo

unds

in h

eat-

trea

ted

food

s (W

oodh

ead

publ

ishi

ng)

Mod

ellin

g of

die

tary

exp

osur

e to

acr

ylam

ide,

Acr

ylam

ide

and

othe

r he

alth

haz

ardo

us c

ompo

unds

in h

eat-

trea

ted

food

s

P17

H. R

. Gla

tt, Y

. Som

mer

In

: Acr

ylam

ide

and

Oth

er H

ealth

H

azar

dous

Com

poun

ds in

Hea

t-tr

eate

d Fo

ods (

K. S

kog,

J. A

lexa

nder

, eds

.),

Woo

dhea

d Pu

blis

hing

, Cam

brid

ge, p

. 32

8-35

7

Hea

lth r

isks

by

5-hy

drox

ymet

hylfu

rfur

al (

HM

F) a

nd r

elat

ed

com

poun

ds

P18

Flur

ina

Cle

men

t, Su

sann

e H

alle

r-B

rem

U

NIM

AG

AZ

IN, U

nive

rsit

y of

Zür

ich

Car

cino

geni

c br

eakf

ast c

erea

ls?

P22

Yea

r 20

07

Skog

K

Food

Sci

ence

and

Tec

hnol

ogy,

21,

25-

27

The

HE

AT

OX

pro

ject

: Hea

t-ge

nera

ted

food

toxi

cant

s: I

dent

ifica

tion

, C

hara

cter

isat

ion

and

Ris

k M

inim

isat

ion

P1

Mur

kovi

c, M

., C

onsu

mer

Pro

tect

ion

thro

ugh

Food

Pr

oces

s Im

prov

emen

t a&

Inn

ovat

ion

in th

e R

eal W

orld

, Ed.

E. L

azos

; Vol

3,

2007

, pp.

502

-510

Tox

ic s

ubst

ance

s in

hea

ted

food

s

P2

of

77

J.S.

Elm

ore

& Q

.Cha

udhr

y D

atab

ase

in M

icro

soft

Acc

ess

The

Hea

tox

data

base

of p

oten

tial

toxi

cant

s re

sulti

ng fr

om li

pid

oxid

atio

n P3

J.S.

Elm

ore

& Q

.Cha

udhr

y D

atab

ase

in M

icro

soft

Acc

ess

The

Hea

tox

data

base

of p

oten

tial

toxi

cant

s re

sulti

ng fr

om th

e M

ailla

rd

reac

tion

P3

J.J.

Kno

l, J.

P.H

. Lin

ssen

and

M

.A.J

.S. v

an B

oeke

l B

i-/t

rien

nial

per

iodi

cal o

n cu

rren

t re

sear

ch o

f the

gra

duat

e sc

hool

VLA

G

Kin

etic

Mod

elin

g of

Acr

ylam

ide

Form

atio

n in

Mod

el S

yste

ms

P7

J.J.

Kno

l Ph

D T

hesi

s, W

agen

inge

n U

nive

rsit

y,

The

Net

herl

ands

K

inet

ic M

odel

ing

of A

cryl

amid

e Fo

rmat

ion

(In

prep

arat

ion)

P7

C. Z

onde

rvan

V

oedi

ngsm

idde

len

Tec

hnol

ogie

(2

007,

in p

repa

rati

on)

Acr

ylam

ide

in N

eder

land

se v

oeds

elpr

oduc

ten

P11

Zon

derv

an, C

.; D

e M

ul, A

. V

oedi

ngsm

idde

lent

echn

olog

ie

Acr

ylam

ide

in v

erhi

tte v

oeds

elpr

oduc

ten:

inna

me

en v

oork

omen

van

in

nam

e (f

inal

title

to b

e co

nfir

med

by

publ

ishe

r). [

Acr

ylam

ide

in h

eate

d fo

od p

rodu

cts:

inta

ke a

nd r

educ

tion

of in

take

]

P11

T B

jella

as

The

sis,

Dis

erta

tion

for

the

degr

ee o

f Ph

iloso

phia

e D

octo

r, U

nive

rsit

y of

O

slo,

200

7

Bio

mar

kers

of d

ieta

ry e

xpos

ure

to a

cryl

amid

e P1

6

Wag

enin

gen

Uni

vers

ity,

The

N

ethe

rlan

ds

Publ

icat

ion:

Bi-

/tri

enni

al p

erio

dica

l on

curr

ent r

esea

rch

of th

e gr

adua

te s

choo

l V

LAG

Hig

her

educ

atio

n In

dust

ry

Res

earc

h

P7

Wag

enin

gen

Uni

vers

ity,

The

N

ethe

rlan

ds

Publ

icat

ion

in p

repa

rati

on: J

ourn

al o

f M

olec

ular

Nut

riti

on a

nd F

ood

Res

earc

h

Hig

her

educ

atio

n R

esea

rch

P7/P

1

Wag

enin

gen

Uni

vers

ity,

The

N

ethe

rlan

ds

Subm

itte

d A

bstr

act:

9th I

nter

nati

onal

Sy

mpo

sium

on

the

Mai

llard

Rea

ctio

n H

ighe

r ed

ucat

ion

Indu

stry

R

esea

rch

P7

of

77

PO

STE

RS

/ OR

AL

P

RE

SEN

TA

TIO

NS

Yea

r 20

03

Aut

hor

Eve

nt

Tit

le

Par

tner

K

erst

in S

kog

E

xper

ts o

n C

onta

min

ants

In

Food

”,

Wor

ksho

p -

Way

s to

Red

uce

Leve

ls o

f A

cryl

amid

e in

Foo

d, B

russ

els,

20-

21

Oct

ober

200

3

“The

HE

AT

OX

pro

ject

” P1

Ker

stin

Sko

g, I

ngeg

erd

Sjöh

olm

W

orks

hop

on A

cryl

amid

e Fo

rmat

ion

in F

ood,

(E

FSA

), 1

7 N

ovem

ber

2003

, B

russ

els.

“The

HE

AT

OX

pro

ject

”

P1

Lilia

Mas

son

Con

fere

ncia

Soc

ieda

d C

hile

na d

e T

ecno

logí

a de

Alim

ento

s Sa

ntia

go, C

hile

“Acr

ilam

ida

En

Alim

ento

s, ¿

Un

Can

ceri

geno

Pot

enci

al”

y “

The

H

EA

TO

X p

roje

ct”

P23

Year

200

4

Ker

stin

Sko

g M

eeti

ng b

efor

e th

e st

art o

f CO

ST 9

27

acti

on, 2

8 Fe

brua

ry 2

004,

Vie

nna.

“T

he H

EA

TO

X p

roje

ct”

P1

Ker

stin

Sko

g N

etw

ork

mee

ting

“A

kryl

amid

i m

aten

”, L

I (S

wed

ish

Food

Ind

ustr

ies)

“T

he H

EA

TO

X p

roje

ct”

P1

Ker

stin

Sko

g ”L

ivsm

edel

sfor

skar

daga

rna”

, G

othe

nbur

g, 1

7 N

ovem

ber

2004

“T

he H

EA

TO

X p

roje

ct”,

P1

Gun

illa

Vik

lund

Li

vsm

edel

sfor

skar

daga

rna,

Göt

ebor

g,

Swed

en

Acr

ylam

ide

in p

otat

o cr

isps

P1

Bag

dona

ite

Öst

erre

ichi

sche

Leb

ensm

itte

lche

mik

er

Tag

e, 1

1.5.

– 1

2.5.

2004

Fo

rmat

ion

of a

cryl

amid

e in

roa

sted

cof

fee

P2

Ahr

né, A

nder

sson

, Flo

berg

, R

osén

, Lin

gner

t Sw

edis

h Fo

od r

esea

rch

Day

s,

Got

henb

urg,

Sw

eden

E

ffec

t of t

empe

ratu

re a

nd w

ater

con

tent

on

acry

lam

ide

form

atio

n du

ring

bak

ing

of w

hite

bre

ad (

Post

er)

P6

Ling

nert

, Han

s JI

FSA

N A

cryl

amid

e U

pdat

e, C

hica

go,

USA

A

cryl

amid

e in

food

– M

itig

atio

n op

tion

s P6

Ling

nert

, Han

s Le

athe

rhea

d K

ey I

ssue

Day

–

Acr

ylam

ide

Up-

date

, Lea

ther

head

, UK

A

cryl

amid

e in

food

– M

itig

atio

n op

tion

s P6

Ling

nert

, Han

s N

ordi

c St

arch

Net

wor

k M

eeti

ng,

Got

henb

urg,

Sw

eden

A

cryl

amid

e an

d st

arch

– S

tate

of t

he a

rt

P6

of

77

Ling

nert

, Han

s C

IAA

acr

ylam

ide

Tec

hnic

al E

xper

t W

orks

hop,

Bru

ssel

s H

EA

TO

X U

pdat

e P6

Ling

nert

, Han

s Sw

edis

h In

dust

ry A

cryl

amid

e N

etw

ork,

M

ay 2

004,

Got

henb

urg

HE

AT

OX

Upd

ate

P6

Ling

nert

, Han

s Sw

edis

h In

dust

ry A

cryl

amid

e N

etw

ork,

O

ctob

er 2

004,

Got

henb

urg

HE

AT

OX

Upd

ate

P6

Ling

nert

, Han

s N

OR

DA

CR

YL

– N

ordi

c N

etw

ork

Proj

ect M

eeti

ng, O

slo,

Nor

way

H

EA

TO

X U

pdat

e P6

Hel

lenä

s, K

.-E

. JR

C-i

rmm

wor

k-sh

op, G

eel,1

7 D

ecem

ber.

T

he H

EA

TO

X p

roje

ct

P9

Dun

ovsk

á L.

, Haj

šlov

á J.

, aj

ka

T.,

Hol

adov

á K

. and

Háj

ková

K.

Che

mic

al R

eact

ions

in F

oods

V.

Prag

ue, C

zech

Rep

ublic

, 29.

thSe

pt -

1.

stO

ct.,

2004

Cha

nges

of A

cryl

amid

e Le

vels

in F

ood

Prod

ucts

dur

ing

Tec

hnol

ogic

al

Proc

essi

ng

P10

J. v

an G

ijsse

l

Pota

to c

onve

ntio

n, J

une

2004

A

mst

erda

m

EU

HE

AT

OX

and

Pot

ato

Proc

essi

ng

P11

E. T

eixi

dó, F

.J. S

anto

s, M

.T.

Gal

cera

n SE

CyT

A (

Mad

rid

2004

) A

naly

sis

of h

ydro

xym

ethy

lfurf

ural

in h

oney

by

gas

chro

mat

ogra

phy

mas

s sp

ectr

omet

ry

P12

Elis

abet

Ber

mud

o, E

ncar

na

Moy

ano,

Lui

s Pu

igno

u, M

aria

T

eres

a G

alce

ran

Reu

nión

Nac

iona

l de

Esp

ectr

omet

ría

de M

asas

LC

-APC

I-M

S/M

S (L

TQ

) fo

r th

e de

term

inat

ion

of a

cryl

amid

e in

fo

odst

uffs

. P1

2

Ele

na B

arce

ló-B

arra

chin

a,

Enc

arna

Moy

ano,

MT

Gal

cera

n

Reu

nión

Nac

iona

l de

Esp

ectr

omet

ría

de M

asas

A

ccur

ate

mas

s m

easu

rem

ent u

sing

a Q

-TO

F fo

r ch

arac

teri

sati

on a

nd

dete

rmin

atio

n of

het

eroc

yclic

am

ines

in fo

od.

P12

H F

rand

sen,

P O

lese

n M

eeti

ng w

ith

Dan

ish

Food

Ind

ustr

y A

cryl

amid

e, r

isk

asse

ssm

ent

P15

Siri

Hel

land

Han

sen

and

Gun

nar

Bru

nbor

g R

epro

duct

ive

Tox

icol

ogy

in th

e A

dult,

R

epro

Safe

Sum

mer

Sch

ool,

Swed

en.

Shor

t pre

sent

atio

n of

the

PhD

pro

ject

- ”

Acr

ylam

ide

in fo

od -

effe

cts o

n re

prod

uctio

n”

P16

H. R

. Gla

tt, W

. Mei

nl, G

. D

obbe

rnac

k, R

. Kol

lock

, U.

Pabe

l, Y

. Som

mer

, et a

l

15th

Int

erna

tiona

l Sym

posiu

m o

n M

icro

som

es a

nd D

rug

Oxi

datio

ns,

Mai

nz, J

uly

2004

Sulp

hotr

ansf

eras

es. (

invi

ted

oral

pre

sent

atio

n)

P18

Y. S

omm

er, H

. Sch

neid

er, H

. R.

Gla

tt

Firs

t Eur

opea

n N

utri

geno

mic

s C

onfe

renc

e, W

agen

inge

n, S

epte

mbe

r 20

04

Haz

ard

iden

tific

atio

n of

the

mut

agen

5-s

ulfo

xym

ethy

l-2-

furf

ural

(S

MF)

form

ed fr

om th

e co

mm

on M

ailla

rd p

rodu

ct 5

-hyd

roxy

met

hyl-

2-fu

rfur

al (

HM

F) b

y in

divi

dual

hum

an s

ulfo

tran

sfer

ases

. (po

ster

)

P18

H. R

. Gla

tt

Sem

inar

s of

Ins

titu

te o

f Pha

rmac

olog

y an

d T

oxic

olog

y, F

acul

ty o

f Vet

erin

ary

Med

icin

e, U

nive

rsit

y of

Gie

ssen

, G

erm

any,

Sep

tem

ber

2004

Sulfo

tran

sfer

asen

als

toxi

fizie

rend

e E

nzym

e [S

ulfo

tran

sfer

ases

as

toxi

fyin

g en

zym

es]

(sem

inar

talk

) P1

8

of

77

H. R

. Gla

tt

Cou

rse

on D

rug

Met

abol

ism

, O

rgan

ized

by

the

Ger

man

Soc

iety

of

Tox

icol

ogy,

in D

ortm

und,

Ger

man

y,

Sept

embe

r 20

04

Sulfo

tran

sfer

asen

und

Hyd

rola

sen

(lec

ture

) P1

8

H. R

. Gla

tt

19th

Eur

opea

n W

orks

hop

on D

rug

Met

abol

ism, A

ntal

ya, T

urke

y, O

ctob

er

2004

Act

ivat

ion

of g

enot

oxic

ants

by

cDN

A-e

xpre

ssed

cyt

ochr

omes

P45

0,

alco

hol d

ehyd

roge

nase

s, a

cety

ltran

sfer

ases

and

sul

fotr

ansf

eras

es:

diff

eren

ces

betw

een

rode

nt a

nd h

uman

enz

yme

form

s (l

ectu

re)

P18

H. R

. Gla

tt

Post

grad

uate

stu

dy c

ours

e on

T

oxic

olog

y, U

nive

rsit

y of

Lei

pzig

, G

erm

any,

Dec

embe

r 20

04

Sulfo

tran

sfer

asen

(le

ctur

e)

P18

H. R

. Gla

tt

Sem

inar

s of

Ins

titu

te fo

r Fo

od

Che

mis

try

and

Env

iron

men

tal

Tox

icol

ogy,

Tec

hnic

al U

nive

rsit

y of

K

aise

rsla

uter

n, G

erm

any,

Dec

embe

r 20

04

Hum

anis

iert

e M

odel

lsys

tem

e zu

m E

rfas

sen

gent

oxis

cher

Wir

kung

en

von

Pyro

lyse

prod

ukte

n un

d N

atur

stof

fen

in L

eben

smit

teln

[H

uman

ized

mod

el s

yste

ms

for

the

dete

ctio

n of

gen

otox

ic e

ffec

ts o

f he

at-i

nduc

ed a

nd n

atur

al fo

od c

onst

itue

nts]

(se

min

ar ta

lk)

P18

Lilia

Mas

son

Con

gres

o N

acio

nal d

e T

ecno

logí

a de

A

limen

tos,

Mar

del

Pla

ta, A

rgen

tina

C

urso

de

Frit

ura

en P

rofu

ndid

ad, M

odul

o 4:

For

mac

ión

de A

crila

mid

a y

“The

HE

AT

OX

pro

ject

” P2

3

Lilia

Mas

son

XII

I Se

min

ario

Lat

inoa

mer

ican

o Y

Del

C

arib

e C

ienc

ia Y

Tec

nolo

gía

De

Alim

ento

s, M

onte

vide

o, U

rugu

ay

“Acr

ilam

ida

En

Alim

ento

s, ¿

Un

Can

ceri

geno

Pot

enci

al”

y “

The

H

EA

TO

X p

roje

ct”

P23

Yea

r 20

05

K S

kog

ILSI

Eur

ope,

wor

ksho

p A

cryl

amid

e ta

sk fo

rce

mee

ting

bra

inst

orm

ing

sess

ion

on r

isk

posi

tion

ing

P1

K S

kog

Nor

dic

Min

istr

y C

ounc

il, M

ölle

(S

wed

en),

8 S

epte

mbe

r, 2

005

“The

HE

AT

OX

pro

ject

”

P1

G V

iklu

nd, K

Ols

son,

I

Sjöh

olm

, K S

kog

16th T

rien

nial

Con

fere

nce

of th

e E

APR

E

ffec

t of s

tora

ge o

n ac

ryla

mid

e in

pot

ato

cris

ps fr

om fi

ve S

wed

ish

grow

n po

tato

var

ieti

es

P1

Skog

K

Frui

t, ve

geta

ble

and

pota

to p

roce

ssin

g,

Bru

gge,

Fo

rmat

ion

of a

cryl

amid

e in

cri

sps

P1

Mur

kovi

c M

iniw

orks

hop

at U

nive

rsit

y of

H

elsi

nki

Form

atio

n of

HM

F P2

of

77

M. M

urko

vic

1st I

nter

nati

onal

Sym

posi

um o

f the

H

uman

Nut

riti

on &

Met

abol

ism

R

esea

rch

and

Tra

inin

gs C

ente

r, G

raz

Aus

tria

Eff

ects

of h

eati

ng o

n fo

od

P2

Pich

ler,

Mur

kovi

c In

vin

o an

alyt

ica

scie

ncia

200

5, 7

.7. –

9.

7.20

05, M

onte

pelli

er, F

ranc

e D

eter

min

atio

n of

HM

F in

win

es a

nd b

alsa

mic

vin

egar

s by

RP-

HPL

C

P2

Der

ler

K.,

Mur

kovi

c M

.

XV

IIth I

nter

nati

onal

Bot

anic

al

Con

gres

s, 1

7.7.

– 2

3.7.

2005

, Wie

n,

Aus

tria

Det

erm

inat

ion

of m

ono

and

disa

ccha

ride

s in

gre

en c

offe

e an

d th

eir

nutr

itio

nal s

igni

fican

ce

P2

Bag

dona

ite,

Mur

kovi

c 8th

Sym

posi

um o

n In

stru

men

tal

Ana

lysi

s, 2

59. –

28.

9.20

05

3-A

PA a

pos

sibl

e in

term

edia

te in

the

path

way

lead

ing

to a

cryl

amid

e P2

Der

ler,

Mur

kovi

c 8th

Sym

posi

um o

n In

stru

men

tal

Ana

lysi

s, 2

59. –

28.

9.20

05

Ana

lysi

s of

am

ino

acid

s an

d ca

rboh

ydra

tes

in g

reen

cof

fee

P2

Mei

Yin

Low

, Don

ald

S.

Mot

tram

and

J. S

teph

en E

lmor

e 11

th W

eurm

an F

lavo

ur R

esea

rch

Sym

posi

um, R

oski

lde,

Den

mar

k

Rel

atio

nshi

p be

twee

n A

cryl

amid

e Fo

rmat

ion

and

the

Gen

erat

ion

of

Flav

our

in H

eate

d Fo

ods

P3

Åm

an P

. N

I m

eeti

ng in

Ice

land

A

cryl

amid

e in

Bre

ad

P4

Mus

tafa

A.

Food

Sci

ence

Day

in U

ppsa

la

Stra

tegi

es to

Red

uce

Acr

ylam

ide

in B

read

P4

M. D

alla

Ros

a W

orks

hop

on b

read

and

bak

ery

prod

ucts

– V

eron

a (I

taly

) –

Ital

ian

Ass

ocia

tion

in F

ood

Tec

hnol

ogy

(AIT

A)

UE

res

earc

h ac

tivi

ty o

n ac

ryla

mid

e an

d to

xica

nts

in h

eat t

reat

ed fo

ods

P5

S. R

oman

i 7th

Ita

lian

Con

gres

s in

Foo

d Sc

ienc

e an

d T

echn

olog

y (C

ISE

TA

) –

Cer

nobb

io (

Ital

y)

Com

pari

son

amon

g di

ffer

ent f

ryer

s on

som

e qu

alit

y ch

arac

teri

stic

s an

d ac

ryla

mid

e co

nten

t of f

ried

pot

atoe

s P5

Ling

nert

Sw

edis

h In

dust

ry A

cryl

amid

e N

etw

ork,

M

arch

200

5, U

ppsa

la

HE

AT

OX

Upd

ate

P6

Ling

nert

Sa

fe F

ood

for

the

Futu

re; Ö

resu

nd

Food

Net

wor

k, L

und,

Sw

eden

A

cryl

amid

e P6

Ling

nert

N

OR

DA

CR

YL

– N

ordi

c N

etw

ork

Proj

ect M

eeti

ng, R

eykj

avik

, Ice

land

H

EA

TO

X U

pdat

e P6

Ling

nert

Sw

edis

h In

dust

ry A

cryl

amid

e N

etw

ork,

O

ctob

er 2

005,

Got

henb

urg

HE

AT

OX

Upd

ate

P6

of

77

Ling

nert

K

SLA

Sem

inar

, Sto

ckho

lm, S

wed

en

Eve

nts

in fo

ods

duri

ng h

eati

ng –

Mai

llard

rea

ctio

n pr

oduc

ts, i

nclu

ding

ac

ryla

mid

e P6

Wag

enin

gen

Uni

vers

ity

Bi-

/tri

enni

al p

erio

dica

l on

curr

ent

rese

arch

of t

he g

radu

ate

scho

ol V

LAG

O

ne-p

age

sum

mar

y of

pro

ject

of p

artn

er 7

P7

J.J.

Kno

l, J.

P.H

. Lin

ssen

and

M

.A.J

.S. v

an B

oeke

l So

crat

es I

P ‘F

ood

& H

ealth

’, In

tern

atio

nal A

dvan

ced

Cou

rse

Tow

ard

a K

inet

ic M

odel

for

Acr

ylam

ide

Form

atio

n in

Mod

el S

yste

ms

(Pos

ter

pres

enta

tion

) P7

J.J.

Kno

l, J.

P.H

. Lin

ssen

and

M

.A.J

.S. v

an B

oeke

l M

eeti

ng w

ith

mem

bers

of D

utch

Po

tato

Pro

cess

ors’

Ass

ocia

tion

(D

PPA

/VA

VI)

Tow

ard

a K

inet

ic M

odel

for

Acr

ylam

ide

Form

atio

n in

Mod

el S

yste

ms

(Pos

ter

pres

enta

tion

) P7

Pete

rsso

n E

.V.,

Ros

én J

., T

urne

r C

., D

anie

lsso

n R

., H

elle

näs

K.-

E.

1st I

nter

nati

onal

Foo

d an

d N

utri

tion

C

ongr

ess,

Ist

anbu

l, T

urke

y, 1

5-18

Ju

ne, 2

005

Cri

tica

l fac

tors

and

pit

falls

aff

ecti

ng th

e ex

trac

tion

of a

cryl

amid

e fr

om

food

s (a

n op

tim

isat

ion

stud

y)

P9

Wag

enin

gen

Uni

vers

ity

Indu

stry

Po

ster

: Mee

ting

wit

h m

embe

rs o

f Dut

ch P

otat

o Pr

oces

sors

’ Ass

ocia

tion

(D

PPA

/VA

VI)

P7

Pete

rsso

n E

.V.,

Ros

én J

., T

urne

r C

., D

anie

lsso

n R

., H

elle

näs

K.E

.

2nd

Inte

rnat

iona

l Sym

posi

um o

n R

ecen

t Adv

ance

s in

Foo

d A

naly

sis,

Pr

ague

, Cze

ch R

epub

lic, 2

-4

Nov

embe

r, 2

005

Iden

tific

atio

n of

cri

tica

l fac

tors

for

extr

acti

on o

f acr

ylam

ide

from

food

s (a

n op

tim

isat

ion

stud

y us

ing

anal

ysis

by

LC-M

S/M

S)

P9

Hel

lenä

s, K

.-E

. SA

NC

O E

xper

t com

mit

tee

on

envi

ronm

enta

l and

indu

stri

al

cont

amin

ants

, Bru

ssel

s, 1

4 Ja

nuar

y.

The

HE

AT

OX

pro

ject

upd

ate.

P9

Hel

lenä

s, K

.-E

. SA

NC

O E

xper

t com

mit

tee

on

envi

ronm

enta

l and

indu

stri

al

cont

amin

ants

, Bru

ssel

s, 9

Jun

e.

Upd

ate

on r

ecen

t HE

AT

OX

act

ivit

ies

on a

cryl

amid

e an

d Fu

ran.

P9

Hel

lenä

s, K

.-E

. B

EU

C m

eeti

ng, B

russ

els,

13

Nov

embe

r.

The

HE

AT

OX

pro

ject

. P9

Hel

lenä

s, K

.-E

. Fo

od s

afet

y in

a E

urop

ean

pers

pect

ive,

Se

min

ar a

nd w

ork-

shop

, Osl

o, 7

-8

Dec

embe

r

Acr

ylam

ide

– St

ill a

mat

ter

of c

once

rn?

P9

Gib

ala

P., N

ovot

ný O

., Pe

ters

son

E.V

., D

unov

ská

L.,

Vác

lavi

k L.

Haj

slov

á J.

, Hel

lenä

s K

.-E

., R

osén

J.

2nd

Inte

rnat

iona

l Sym

posi

um o

n R

ecen

t Adv

ance

s in

Foo

d A

naly

sis,

Pr

ague

, Cze

ch R

epub

lic, 2

-4

Nov

embe

r, 2

005

Can

alk

alin

e ex

trac

tion

rel

ease

mas

ked

acry

lam

ide?

P9

P1

0

of

77

Gib

ala

P., P

eter

sson

E.,

Dun

ovsk

á L.

, Vác

laví

k L.

, N

ovot

ný O

., H

ajšl

ová

J.,

Hel

lenä

s K

. E.,

Ros

én J

.

2nd I

nter

nati

onal

sym

posi

um o

n re

cent

ad

vanc

es in

food

ana

lysi

s, P

ragu

e,

Cze

ch R

epub

lic, 2

-4 N

ovem

ber,

200

5

Can

alk

alin

e ex

trac

tion

rel

ease

mas

ked

acry

lam

ide?

P1

0

Dun

ovsk

á L.

, aj

ka T

. and

H

ajšl

ová

J.

3rd M

eeti

ng o

n C

hem

istr

y &

Life

Brn

o,

Cze

ch R

epub

lic, 2

0-22

Sep

tem

ber,

20

05

Acr

ylam

ide

Leve

ls in

Foo

dstu

ffs

from

Cze

ch m

arke

t P1

0

Dun

ovsk

á L.

, Haj

šlov

á J.

, H

olad

ová

K.,

Gib

ala

P., S

chre

k J.

2nd I

nter

nati

onal

sym

posi

um o

n re

cent

ad

vanc

es in

food

ana

lysi

s, P

ragu

e,

Cze

ch R

epub

lic, 2

-4 N

ovem

ber,

200

5

Ana

lyti

cal s

trat

egie

s fo

r ex

amin

atio

n of

roa

sted

cof

fee:

vol

atile

mar

kers

of

acr

ylam

ide

form

atio

n P1

0

J. v

an G

ijsse

l

Acr

ylam

ide

stak

ehol

ders

mee

ting

, 14

Janu

ary

2005

, Bru

ssel

H

EA

TO

X W

P 2.

For

mat

ion

and

Proc

essi

ng, P

rogr

ess

in th

e fir

st y

ear.

P1

1

E. B

erm

udo,

O. N

úñez

, L.

Puig

nou,

M.T

. Gal

cera

n X

I Jo

rnad

as d

e A

nális

is I

nstr

umen

tal

(Bar

celo

na 2

005)

D

evel

opm

ent o

f new

cap

illar

y el

ectr

opho

resi

s m

etho

dolo

gies

for

the

anal

ysis

of a

cryl

amid

e in

food

stuf

fs

P12

E. T

eixi

dó, F

.J. S

anto

s, M

.T.

Gal

cera

n X

I Jo

rnad

as d

e A

nális

is I

nstr

umen

tal

(Bar

celo

na 2

005)

G

as c

hrom

atog

raph

y co

uple

d to

mas

s sp

ectr

omet

ry fo

r th

e an

alys

is o

f hy

drox

ymet

hylfu

rfur

al in

food

s P1

2

M. S

. Alta

ki, F

.J. S

anto

s, M

.T.

Gal

cera

n X

I Jo

rnad

as d

e A

nális

is I

nstr

umen

tal

(Bar

celo

na 2

005)

A

naly

sis

of fu

ran

in fo

od b

y he

adsp

ace

solid

-pha

se m

icro

extr

acti

on

coup

led

wit

h ga

s ch

rom

atog

raph

y-m

ass

spec

trom

etry

. P1

2

Elis

abet

Ber

mud

o, O

scar

Núñ

ez,

Luis

Pui

gnou

, Mar

ia T

eres

a G

alce

ran

HPL

C 2

005

Cap

illar

y Z

one

Ele

ctro

phor

esis

for

the

dete

rmin

atio

n of

acr

ylam

ide

in

food

stuf

fs

P12

Ele

na B

arce

ló-B

arra

chin

a,

Enc

arna

ción

Moy

ano,

Llu

ís

Puig

nou

and

Mar

ia T

eres

a G

alce

ran

HPL

C 2

005

Dev

elop

men

t of m

etha

cryl

ate

mon

olit

hs fo

r th

e an

alys

is o

f het

eroc

yclic

am

ines

by

capi

llary

ele

ctro

chro

mat

ogra

phy

P12

Mar

ia T

eres

a G

alce

ran

1st

Int

erna

tion

al W

orks

hop

of M

S/M

S sp

ectr

omet

ry

Tan

dem

Mas

s Sp

ectr

omet

ry a

nd A

ccur

ate

Mas

s M

easu

rem

ent f

or th

e an

alys

is o

f gen

otox

ic h

eat-

indu

ced

com

poun

ds in

food

. P1

2

Hül

ya Ö

lmez

T

ÜB

TA

K M

RC

1st

Int

erna

tion

al

Food

and

Nut

riti

on C

ongr

ess-

15-1

8 Ju

ne 2

005,

Ist

anbu

l / O

ral

pres

enta

tion

Gıd

alar

da A

krila

mid

(A

cryl

amid

e in

food

s)

P13

Hül

ya Ö

lmez

T

V P

rogr

amm

e -

AT

V C

hann

el /

sp

eake

r A

cryl

amid

e ri

sk in

food

s P1

3

of

77

Hül

ya Ö

lmez

T

V P

rogr

amm

e –

CN

BC

-e C

hann

el /

sp

eake

r A

cryl

amid

e ri

sk in

food

s P1

3

P O

lese

n O

ral p

rese

ntat

ion

inte

rnal

ly in

DFV

F A

kryl

amid

– a

naly

se a

f glo

bina

dduk

ter

I hu

man

e pr

øver

(pr

ojec

t H

EA

TO

X)

P15

Tho

mas

Bje

llaas

N

orw

egia

n So

ciet

y fo

r M

ass

Spec

trom

etry

, Haf

jell

Janu

ary

2005

Po

ster

– “

Det

erm

inat

ion

of a

cryl

amid

e m

etab

olit

es in

hum

an u

rine

us

ing

solid

-pha

se e

xtra

ctio

n an

d LC

-MS/

MS

dete

ctio

n”

P16

Tho

mas

Bje

llaas

, Heg

e B

. Ø

lstø

rn, J

an E

rik

Paul

sen

Nor

dic

mee

ting

for

NO

RD

AC

RY

L –

june

, Ice

land

Pr

esen

tati

on o

f bio

avai

labi

lity

of a

cryl

amid

e fr

om fo

ods

stud

y in

mic

e us

ing

met

abol

ic c

ages

P1

6

Tho

mas

Bje

llaas

E

urop

ean

mee

ting

for

HE

AT

OX

–

(Net

herl

ands

), M

ay 2

005

Post

er –

“D

eter

min

atio

n of

acr

ylam

ide

met

abol

ites

in h

uman

uri

ne

usin

g so

lid-p

hase

ext

ract

ion

and

LC-M

S/M

S de

tect

ion”

P1

6

Tho

mas

Bje

llaas

N

orw

egia

n In

stit

ute

of P

ublic

Hea

lth

Pres

enta

tion

– A

pilo

t stu

dy o

n ur

inar

y ex

cret

ion

acry

lam

ide

met

abol

ites

in h

uman

s P1

6

Heg

e B

. A. Ø

lstø

rn, J

an E

rik

Paul

sen,

Jan

Ale

xand

er

Nor

weg

ian

Inst

itut

e of

Pub

lic H

ealth

Pr

esen

tati

on o

f pre

limin

ary

resu

lts –

Acr

ylam

ide

and

inte

stin

al c

ance

r P1

6

Siri

Hel

land

Han

sen,

Eri

k Sø

derl

und,

Ado

lf B

aum

gart

ner,

D

And

erso

n an

d G

Bru

nbor

g

3rd C

open

hage

n W

orks

hop

on

Env

iron

men

t, R

epro

duct

ive

Hea

lth a

nd

Fert

ility

, 15-

18 J

anua

ry 2

005

Post

er a

nd a

bstr

act –

“T

he p

ater

nal r

epro

duct

ive

toxi

city

of t

he fo

od

cont

amin

ant a

cryl

amid

e P1

6

Siri

Hel

land

Han

sen

and

Gun

nar

Bru

nbor

g N

orw

egia

n In

stit

ute

of P

ublic

Hea

lth,

Div

isio

n of

Env

iron

men

tal M

edic

ine

Pres

enta

tion

(in

Nor

weg

ian)

– “

Kan

akr

ylam

id p

åvir

ke d

en m

annl

ige

repr

oduk

sjon

en?”

P1

6

Jan

Ale

xand

er

9th I

nter

nati

onal

Con

fere

nce

on

Env

iron

men

tal M

utag

ens,

San

Fr

anci

sco,

Sep

t. 20

05

Ris

k A

sses

smen

t of A

cryl

amid

e in

Foo

d P1

6

Jaco

b D

. van

Kla

vere

n H

eato

x 18

th m

onth

mee

ting

, Sy

mpo

sium

wit

h D

utch

pot

ato

proc

esso

rs A

ssoc

iati

on

Exp

osur

e as

sess

men

t of H

EA

TO

X c

ompo

unds

; P1

7

Jaco

b D

. van

Kla

vere

n, A

nika

de

Mul

D

utch

Foo

d A

utho

rity

(V

WA

) w

ork-

mee

ting

E

xpos

ure

asse

ssm

ent o

f hea

t-tr

eate

d to

xica

nts

P17

Ani

ka d

e M

ul

Food

Sta

ndar

d A

genc

y (U

K)

, wor

k-m

eeti

ng

Prob

abili

stic

acr

ylam

ide

inta

ke c

alcu

lati

ons

wit

h M

CR

A a

nd

Com

pari

son

of a

cryl

amid

e in

take

by

Bri

tish

and

Dut

ch to

ddle

rs

P17

H. R

. Gla

tt

Sym

posi

um o

f Sem

inar

s of

B

unde

sins

titu

t für

Ris

ikob

ewer

tung

([

Ger

man

] Fe

dera

l Ins

titu

te o

f Ris

k A

sses

smen

t), F

ebru

ary

2005

Gen

toxi

zitä

ts-U

nter

such

unge

n in

vitr

o m

it v

ersc

hied

enen

K

ohle

nhyd

rat-

Pyro

lyse

prod

ukte

n [G

enot

oxic

ity

stud

ies

in v

itro

on

vari

ous

carb

ohyd

rate

pyr

olys

is p

rodu

cts]

(ta

lk)

P18

of

77

H. R

. Gla

tt

Ann

ual M

eeti

ng o

f the

“A

rbei

tskr

eis

Ges

undh

eit

und

Lebe

nsm

itte

lsic

herh

eit”

[w

ork

grou

p H

ealth

and

Foo

d Sa

fety

] of

the

“Bun

desv

erba

ndes

der

Deu

tsch

en

Süßw

aren

indu

stri

e e.

V.”

[Fe

dera

l A

ssoc

iati

on o

f the

Ger

man

Ind

ustr

y on

Sw

eet P

rodu

cts]

, Ber

ghol

z-R

ehbr

ücke

, G

erm

any,

Jun

e 20

05

Hea

tox:

G

ento

xizi

täts

-Unt

ersu

chun

gen

mit

ve

rsch

iede

nen

Koh

lenh

ydra

t-Py

roly

sepr

oduk

ten

[Hea

tox:

G

enot

oxic

ity

stud

ies

on

vari

ous

carb

ohyd

rate

pyr

olys

is p

rodu

cts]

(ta

lk)

P18

H. R

. Gla

tt

Sem

inar

s of

the

Inst

itut

e fo

r C

ance

r R

esea

rch,

Uni

vers

ity

of V

ienn

a,

Aus

tria

, Jun

e 20

05

Gen

toxi

sche

und

kan

zero

gene

Wir

kung

en v

on L

eben

smit

tel-

Inha

ltsst

offe

n in

hum

anis

iert

en M

odel

lsys

tem

en u

nd d

eren

Pr

även

tion

/Pot

enzi

erun

g du

rch

wei

tere

Ern

ähru

ngsf

akto

ren

[Gen

otox

ic a

nd c

arci

noge

nic

effe

cts

of fo

od-b

orne

sub

stan

ces

in

hum

aniz

ed m

odel

sys

tem

s, a

nd th

eir

prev

enti

on/p

oten

tiat

ion

by o

ther

fo

od fa

ctor

s] (

sem

inar

talk

)

P18

H. R

. Gla

tt

Post

grad

uate

stu

dy c

ours

e on

T

oxic

olog

y, U

nive

rsit

y of

Vie

nna,

A

ustr

ia, J

une

2005

Gen

etis

ch v

erän

dert

e Z

elle

n un

d T

iere

zur

Unt

ersu

chun

g de

s Fr

emds

toff

-Met

abol

ism

us [

Gen

etic

ally

eng

inee

red

cells

and

ani

mal

s fo

r in

vest

igat

ions

on

xeno

biot

ic m

etab

olis

m]

(lec

ture

)

P18

H. R

. Gla

tt

Con

tinu

ous

Edu

cati

on in

Tox

icol

ogy,

Sy

mpo

sium

„R

ücks

tand

stox

ikol

ogie

, ne

ue A

spek

te u

nd E

ntw

ickl

unge

n 20

05“

Uni

vers

ity

of Z

uric

h,

Swit

zerl

and,

Nov

embe

r 20

05

Gen

otox

isch

e Py

roly

sepr

oduk

te u

nd N

atur

stof

fe in

Leb

ensm

itte

ln

[Gen

otox

c py

roly

sis

prod

ucts

and

nat

ural

com

poun

ds in

food

s]

(lec

ture

)

P18

Bar

bara

Gal

lani

B

EU

C F

ood

offic

ers’

mee

ting

29

-30

Nov

embe

r 20

05

Acr

ylam

ide:

reg

ulat

ory

appr

oach

and

com

mun

icat

ion

to c

onsu

mer

s P2

0

Hel

ga O

dden

Rek

snes

Fo

od p

ract

ices

at h

ome;

wha

t are

the

risk

s?

Wor

ksho

ps I

rela

nd a

nd N

orth

ern

Irel

and

17 a

nd 1

8 M

ay 2

005

The

rol

e of

ris

k co

mm

unic

atio

n po

licy

in E

urop

e in

ass

urin

g fo

od

safe

ty to

con

sum

ers

(man

uscr

ipt a

nd p

pt-p

rese

ntat

ion)

P21

Hel

ga O

dden

Rek

snes

D

G S

anco

, CC

AB

mee

ting

H

EA

TO

X –

Del

iver

able

s –

Rsi

k C

omm

unic

atio

n an

d di

ssem

inat

ion

(ppt

-pre

sent

atio

n)

P21

Han

spet

er N

aege

li C

ours

e in

Tox

icol

ogy

Swis

s Fe

dera

l Ins

titu

te o

f Tec

hnol

ogy

The

app

licat

ion

of “

-om

ic”

tech

nolo

gies

in fo

od to

xico

logy

P2

2

Flur

ina

Cle

men

t In

tern

atio

nal S

ympo

sium

of

Phar

mac

olog

y an

d T

oxic

olog

y. V

ienn

a G

ene

expr

essi

on p

rofil

es in

duce

d by

gly

cida

mid

e in

hum

an c

ells

P2

2

of

77

Flur

ina

Cle

men

t A

nnua

l Mee

ting

of t

he S

wis

s T

oxic

olog

y N

etw

ork

(XE

RR

) G

ene

expr

essi

on p

rofil

es in

duce

d by

gly

cida

mid

e in

hum

an c

ells

P2

2

Flur

ina

Cle

men

t A

nnua

l Mee

ting

of t

he X

enob

ioti

c an

d E

nvir

onm

enta

l Ris

k R

esea

rch

Cen

ter

Zür

ich,

Sw

itze

rlan

d

Tra

nscr

ipto

mic

s of

Acr

ylam

ide

and

Gly

cida

mid

e P2

2

Lilia

Mas

son

VI

Sem

inar

io L

atin

oam

eric

ano

de

Cie

ncia

de

Alim

ento

s, C

ampi

nas,

B

rasi

l

“Acr

ilam

ida

En

Alim

ento

s, ¿

Un

Can

ceri

geno

Pot

enci

al”

y “

The

H

EA

TO

X p

roje

ct”

P23

Lilia

Mas

son

Con

gres

o Fe

dera

ción

Far

mac

éuti

ca

Suda

mer

ican

a, S

anti

ago,

Chi

le

“Acr

ilam

ida

En

Alim

ento

s, ¿

Un

Can

ceri

geno

Pot

enci

al”

y “

The

H

EA

TO

X p

roje

ct”

P23

Year

200

6

Skog

K

Env

iron

men

tal a

nd I

ndus

tria

l C

onta

min

ants

wor

ksho

p- a

cryl

amid

e,

(CIA

A)

Bru

ssel

s

The

HE

AT

OX

pro

ject

- H

eat –

gen

erat

ed F

ood

Tox

ican

ts.

Iden

tific

atio

n, C

hara

cter

izat

ion

and

Ris

k M

inim

isat

ion

Skog

K

HE

AT

OX

wor

ksho

p, G

raz

Hom

e-co

okin

g an

d ac

ryla

mid

e

P1

Skog

K

Wor

ksho

p at

Nes

tlé, L

ausa

nne

The

HE

AT

OX

pro

ject

P1

Skog

K

Form

as v

isit

to L

U

Bild

ning

av

toxi

ska

ämne

n vi

d liv

smed

elsp

roce

ssni

ng

Bild

ning

smek

anis

mer

och

häl

soef

fekt

er

P1

G V

iklu

nd, K

Ols

son,

I

Sjöh

olm

, K S

kog

Livs

med

elsf

orsk

arda

garn

a, N

ovem

ber

21-

22, 2

006,

Upp

sala

, Sw

eden

A

cryl

amid

e in

cri

sps

from

Sw

edis

h gr

own

Satu

rna

pota

toes

P1

G V

iklu

nd, K

Ols

son,

I

Sjöh

olm

, K S

kog

Mee

ting

NO

RD

AC

RY

L E

ffec

t of s

tora

ge o

n ac

ryla

mid

e in

pot

ato

cris

ps fr

om fi

ve S

wed

ish

grow

n po

tato

var

ieti

es

P1

Bag

dona

ite

K, M

urko

vic

M

Ö

ster

reic

hisc

he L

eben

smit

telc

hem

iker

T

age,

12.

9. –

14.

9.20

06, W

ien

Acr

ylam

ide

form

atio

n in

cof

fee

P2

Bor

nik

MA

, Jöb

stl D

, Der

ler

K,

Mur

kovi

c M

Öst

erre

ichi

sche

Leb

ensm

itte

lche

mik

er

Tag

e, 1

2.9.

– 1

4.9.

2006

, Wie

n K

inet

ik d

er H

MF-

und

HM

FS-B

ildun

g im

Lau

fe d

er K

affe

erös

tung

, in

Mod

ells

yste

men

P2

Per

Åm

an

Arr

ange

d a

mee

ting

in c

olla

bora

tion

w

ith

Nor

dic

Inno

vati

on C

entr

e (N

ICe

Mee

ting

NO

RD

AC

RY

L)

Acr

ylam

ide

P4

of

77

Arw

a M

usta

fa

NIC

e M

eeti

ng N

OR

DA

CR

YL

Ana

lysi

s of

Fre

e A

min

o A

cids

P4

Arw

a M

usta

fa

Lect

ure

at S

udan

Uni

vers

ity,

K

hart

oum

, Sud

an

Acr

ylam

ide

in F

ood

Prod

ucts

P4

Arw

a M

usta

fa

Lect

ure

at A

hfad

Uni

vers

ity,

O

mdu

rman

, Sud

an

Acr

ylam

ide

in F

ood

Prod

ucts

P4

M. D

alla

Ros

a C

IAA

Bru

ssel

les

Mee

ting

H

eat –

gen

erat

ed fo

od to

xica

nts.

Ide

ntifi

cati

on, c

hara

cter

izat

ion

and

risk

min

imis

atio

n. H

ome

cook

ing

P5

S. R

oman

i, M

. Bac

chio

cca,

P.

Roc

culi,

A. A

ngio

loni

, M. D

alla

R

osa

CE

Food

Con

gres

s: 3

th C

entr

al

Eur

opea

n C

ongr

ess

on F

ood.

(So

fia -

B

ulga

ria)

Eff

ect o

f fry

ing

tim

e on

som

e qu

alit

y as

pect

s an

d ac

ryla

mid

e co

nten

t of

Fren

ch f

ries

P5

S. R

oman

i, M

. Bac

chio

cca,

P.

Roc

culi,

M. D

alla

Ros

a IU

FOST

, 13th

Wor

ld C

ongr

ess

of

Food

Sci

ence

and

Tec

hnol

ogy

(Nan

tes

– Fr

ance

)

Eff

ect o

f som

e fr

ying

con

diti

ons

on a

cryl

amid

e co

nten

t in

fren

ch fr

ies

P5

Ling

nert

, Han

s Sw

edis

h In

dust

ry A

cryl

amid

e N

etw

ork,

M

arch

200

6, U

ppsa

la

HE

AT

OX

Upd

ate

P6

Ling

nert

, Han

s H

EA

TO

X W

orks

hop,

Jun

e 20

06,

Gra

z, A

ustr

ia

Min

imis

atio

n O

ptio

ns -

Ind

ustr

y P6

Ling

nert

, Han

s

CIA

A A

cryl

amid

e G

roup

, 5 J

uly

2006

, B

russ

els,

Bel

gum

H

EA

TO

X d

eliv

erab

les

in r

elat

ion

to th

e C

IAA

Too

lbox

P6

Ling

nert

, Han

s Sw

edis

h In

dust

ry A

cryl

amid

e N

etw

ork,

O

ctob

er 2

006,

Got

henb

urg

HE

AT

OX

Upd

ate

P6

Hel

lenä

s, K

.-E

. SA

NC

O/C

IAA

Wor

ksho

p, B

russ

els,

16

Mar

ch.

Acr

ylam

ide

extr

acti

on a

t hig

h pH

. P9

Ling

nert

, Han

s Sw

edis

h Fo

od R

esea

rch

Day

s, U

ppsa

la,

21-2

2 N

ovem

ber

2006

A

kryl

amid

– p

å go

tt o

ch o

nt

Vác

laví

k L.

, Bar

táko

vá V

., H

olad

ová

K.,

Haj

šlov

á J.

JA

KO

ST O

BIL

OV

IN -

200

6 (Q

ualit

y of

cer

eals

-200

6),

Kro

míž

, C

zech

Rep

ublic

, 9.1

1.20

06

Acr

ylam

ide

– ne

w fo

od c

arci

noge

n P1

0

Haj

šlov

á J.

, Nov

otný

O.,

Vác

laví

k L,

Dun

ovsk

á L.

C

OST

IM

AR

S Jo

int W

orks

hop,

N

apol

i, It

aly,

24.

- 2

7. 5

. 200

6 St

udy

of "

mas

ked"

acr

ylam

ide

orig

in

P10

Vác

laví

k L,

aj

ka T

., D

unov

ská

L., H

ajšl

ová

J.

CO

ST I

MA

RS

Join

t Wor

ksho

p,

Nap

oli,

Ital

y, 2

4. -

27.

5. 2

006

LC-M

S/M

S an

d G

C/H

RT

OF-

MS

met

hods

for

anal

ysis

of a

cryl

amid

e in

food

stuf

fs

P10

Vác

laví

k L.

, Bar

táko

vá V

., H

olad

ová

K.,

Haj

šlov

á J.

JA

KO

ST O

BIL

OV

IN -

200

6 (Q

ualit

y of

cer

eals

-200

6),

A

cryl

amid

e –

new

food

car

cino

gen

P10

of

77

Zon

derv

an, C

. W

orks

hop

wit

h D

PPA

N

ov 2

8, 2

006

Inst

ruct

ions

on

usin

g th

e fr

ench

fry

mod

ellin

g to

ol: F

ry S

imul

ator

v.2

P1

1

E. T

eixi

dó, E

. Moy

ano,

F.J

. Sa

ntos

, M.T

. Gal

cera

n II

I R

euni

ón d

e la

Soc

ieda

d E

spañ

ola

de

Esp

ectr

omet

ría

de M

asas

(O

vied

o 20

06)

Ana

lysi

s of

hyd

roxy

met

hylfu

rfur

al in

food

stuf

fs b

y liq

uid

chro

mat

ogra

phy

coup

led

to ta

ndem

mas

s sp

ectr

omet

ry

P12

L. P

uign

ou

Jorn

ada

de G

enot

òxic

s en

Alim

ents

(R

eus

2006

) T

óxic

os a

limen

tari

os e

ndog

énic

os e

n al

imen

tos

proc

esad

os

térm

icam

ente

P1

2

E. B

erm

udo

Jorn

ada

de G

enot

òxic

s en

Alim

ents

(R

eus

2006

) A

nális

is d

e ac

rila

mid

a en

alim

ento

s P1

2

E. T

eixi

dó

Jorn

ada

de G

enot

òxic

s en

Alim

ents

(R

eus

2006

) A

nális

is d

e hi

drox

imet

ilfur

fura

l med

iant

e té

cnic

as c

rom

atog

ráfic

as y

es

pect

rom

etrí

a de

mas

as

P12

M. S

. Alta

ki

Jorn

ada

de G

enot

òxic

s en

Alim

ents

(R

eus

2006

) Fu

ran:

For

mat

ion

and

anal

ysis

P1

2

E. B

erm

udo,

O. N

úñez

, E.

Moy

ano,

L. P

uign

ou, M

.T.

Gal

cera

n

SEC

yTA

(V

igo

2006

) A

pplic

atio

n of

ele

ctro

phor

etic

tech

niqu

es fo

r th

e de

term

inat

ion

of

acry

lam

ide

in fo

ods

P1

2

M. S

. Alta

ki, F

.J. S

anto

s, M

.T.

Gal

cera

n SE

CyT

A (

Vig

o 20

06)

Ana

lysi

s of

fura

n in

food

by

on-l

ine

head

spac

e so

lid-p

hase

m

icro

extr

acti

on a

nd g

as c

hrom

atog

raph

y io

n tr

ap m

ass

spec

trom

etry

. P1

2

A V

ikst

röm

, S E

riks

son,

B

Paul

sson

, P K

arls

son

and

M

Tör

nqvi

st

Pres

enta

tion

at Å

bo A

cade

my

Stud

y of

rel

ativ

e bi

oava

ilabi

lity

of a

cryl

amid

e in

diff

eren

t foo

dstu

ffs

P14

A V

ikst

röm

Se

min

ar a

t Sto

ckho

lm U

nive

rsit

y B

ackg

roun

d ex

posu

re to

acr

ylam

ide

and

its

geno

toxi

c m

etab

olit

e gl

ycid

amid

e P1

4

A V

ikst

röm

, B P

auls

son,

& M

T

örnq

vist

C

ours

e at

Kar

olin

ska

Inst

itut

et,

Soln

a,Sw

eden

(E

CN

IS),

pos

ter

pres

enta

tion

Poss

ible

influ

ence

of p

olym

orph

ism

on

inte

rnal

dos

e of

acr

ylam

ide

and

glyc

idam

ide

P14

M T

örnq

vist

H

EA

TO

X w

orks

hop

13-1

4 Ju

ne

The

acr

ylam

ide

stor

y P1

4 A

nika

de

Mul

IL

SI, E

xper

t gro

up m

eeti

ng ‘R

isk

Ben

efit

Ana

lysi

s of

Mit

igat

ion

Mea

sure

s’

Acr

ylam

ide;

Exp

osur

e an

d R

educ

tion

sce

nari

os

P17

Jaco

b D

. van

Kla

vere

n N

orw

egai

n Fo

od A

utho

rity

(V

KM

) Pr

obab

ilist

ic m

odel

ing

in p

ersp

ecti

ve o

f fut

ure

risk

ass

essm

ent

P17

Jaco

b D

. van

Kla

vere

n N

aam

van

het

sym

posi

um. T

he

HE

AT

OX

Wor

ksho

p

Exp

osur

e an

d R

educ

tion

Sce

nari

os

P17

of

77

Jaco

b D

. van

Kla

vere

n T

elev

isio

n pr

ogra

mm

e fo

r co

nsum

ers

(Kas

sa)

Acr

ylam

ide

leve

ls in

food

and

its

risk

s P1

7

H. R

. Gla

tt

Con

tinu

ous

Edu

cati

on fo

r M

edic

al

Doc

tors

, Cou

rse

“Ern

ähru

ngs-

med

izin

”, P

otsd

am-R

ehbr

ücke

, G

erm

any,

Mar

ch 2

006

Kre

bser

zeug

ende

Sto

ffe

in L

eben

smit

teln

[Fo

od-b

orne

car

cino

gens

] (l

ectu

re)

P18

H. R

. Gla

tt

Cou

rse

on D

rug

Met

abol

ism

, O

rgan

ized

by

the

Ger

man

Soc

iety

of

Tox

icol

ogy,

in D

ortm

und,

Ger

man

y,

Sept

embe

r 20

06

Sulfo

tran

sfer

asen

und

Hyd

rola

sen

(lec

ture

) P1

8

H. R

. Gla

tt, Y

. Som

mer

, G.

Dob

bern

ack,

W. M

einl

43

rd C

ongr

ess o

f the

Eur

opea

n So

ciet

ies

of T

oxic

olog

y an

d 6t

h C

ongr

ess o

f T

oxic

olog

y in

Dev

elop

ing

Cou

ntri

es,

Cav

tat/

Dub

rovn

ik, C

roat

ia, S

epte

mbe

r

Het

erol

ogou

s an

d tr

ansg

enic

mod

els

for

stud

ying

gen

otox

ic e

ffec

ts o

f co

ntam

inan

ts p

rodu

ced

by h

eat-

trea

tmen

t of f

ood

(inv

ited

talk

),

Abs

trac

t pub

lishe

d in

: Tox

icol

. Let

t. 16

4 S

(200

6) 6

2-63

.

P18

Jaco

b D

. van

Kla

vere

n T

rain

ing

for

Nor

weg

ian

Food

A

utho

rity

(V

KM

) Pr

obab

ilist

ic d

ieta

ry in

take

cal

cula

tion

s fo

r ac

ryla

mid

e P1

7

H. R

. Gla

tt, F

. Tau

gner

, S.

Flor

ian,

Y. S

omm

er

CO

ST 9

26/9

27 C

onfe

renc

e:

Inte

rnat

iona

l Con

fere

nce

on M

olec

ular

an

d Ph

ysio

logi

cal E

ffect

s of B

ioac

tive

Food

Com

pone

nts,

Vie

nna,

Aus

tria

, O

ctob

er 2

006

Hea

lth r

isks

by

5-hy

drox

ymet

hylfu

rfur

al (

HM

F) a

nd r

elat

ed

com

poun

ds (

invi

ted

talk

), S

ome

data

acc

essi

ble

at C

OST

926

web

pa

ge: h

ttp:

//w

ww

.uoc

hb.c

as.c

z/Z

prav

y/C

OST

_926

/

P18

H. R

. Gla

tt, G

. Dob

bern

ack,

W.

Mei

nl

CO

ST 9

26/9

27 C

onfe

renc

e:

Inte

rnat

iona

l Con

fere

nce

on M

olec

ular

an

d Ph

ysio

logi

cal E

ffect

s of B

ioac

tive

Food

Com

pone

nts,

Vie

nna,

Aus

tria

, O

ctob

er 2

006

Exp

ress

ion

of h

uman

pha

se-I

I en

zym

es in

mod

el s

yste

ms

for

dete

ctin

g ge

noto

xica

nts

and

prot

ecti

ve a

gent

s (i

nvit

ed ta

lk),

Som

e da

ta a

cces

sibl

e at

CO

ST92

6 w

eb p

age:

htt

p://

ww

w.u

ochb

.cas

.cz/

Zpr

avy/

CO

ST_9

26/

P18

Bar

bara

Gal

lani

C

IAA

/EC

Wor

ksho

p 16

-17

Mar

ch 2

006

Acr

ylam

ide:

info

rmat

ion

and

com

mun

icat

ion

to c

onsu

mer

s P2

0

Hel

ga O

dden

Rek

snes

A

cryl

amid

e W

orks

hop

EU

/CIA

A

Min

utes

from

wor

king

gro

up o

n ho

me

cook

ing

P21

Hel

ga O

dden

Rek

snes

C

CFA

C W

orks

hop

on R

isk

com

mun

icat

ion

as a

man

agem

ent t

ool

for

cont

amin

ants

Con

sum

ptio

n A

dvic

e on

Con

tam

inan

ts

- a

brie

f com

mun

icat

ion

case

stu

dy o

n A

cryl

amid

e

P21

Wor

king

Gro

up, H

EA

TO

X

Wor

ksho

p

The

HE

AT

OX

Wor

ksho

p R

epor

t P2

1

of

77

Hel

ga O

dden

Rek

snes

T

he H

EA

TO

X W

orks

hop

The

HE

AT

OX

Aqu

ariu

m

P21

Hel

ga O

dden

Rek

snes

T

he H

EA

TO

X 3

0 M

mee

ting

Gra

z Su

mm

ary

Wor

ksho

p G

raz

P21

Flur

ina

Cle

men

t M

eeti

ng o

f the

DN

A R

epai

r-R

eplic

atio

n-R

ecom

bina

tion

Net

wor

k Z

üric

h, S

wit

zerl

and

Tra

nscr

ipto

mic

s of

Gly

cida

mid

e, th

e G

enot

oxic

Met

abol

ite

of

Acr

ylam

ide

P22

Year

200

7

G V

iklu

nd, K

Ols

son,

I

Sjöh

olm

, K S

kog

4th E

APR

/FN

K/U

EIT

P Po

tato

Pr

oces

sing

Con

fere

nce

and

EA

PR

Eng

inee

ring

& U

tilis

atio

n Se

ctio

n M

eeti

ng, J

anua

ry 1

7, 2

007,

Gra

ntha

m,

UK

Acr

ylam

ide

in c

risp

s fr

om S

wed

ish

grow

n Sa

turn

a po

tato

es

P1

G V

iklu

nd, K

Ols

son,

I

Sjöh

olm

, K S

kog

Firs

t Eur

opea

n W

orks

hop

on F

ood

Eng

inee

ring

and

Tec

hnol

ogy,

EFF

oST

E

FChE

Foo

d W

orki

ng P

arty

Con

gres

s,

May

200

7, B

erlin

, Ger

man

y

Acr

ylam

ide

in p

otat

o cr

isps

from

Sw

edis

h gr

own

pota

toes

P1

G V

iklu

nd, K

Ols

son,

I

Sjöh

olm

, K S

kog

CO

ST 9

27 A

ctio

n W

orks

hop,

Sof

ia,

Bul

gari

a A

cryl

amid

e in

roa

sted

pot

ato

wed

ges

P1

Skog

K

Sym

posi

um o

n C

hem

istr

y an

d T

oxic

olog

y of

Acr

ylam

ide,

Bos

ton,

US,

A

ugus

t, 20

07.

Hea

t der

ived

toxi

cant

s in

food

– s

ome

findi

ngs

of a

col

labo

rati

ve

Eur

opea

n re

sear

ch p

roje

ct

P1

Mur

kovi

c, M

. 5th

Int

erna

tion

al C

ongr

ess

on F

ood

Tec

hnol

ogy,

9.3

.-11

.3.2

007

The

ssal

onik

i/G

reec

e

Form

atio

n of

pot

enti

ally

toxi

c su

bsta

nces

dur

ing

heat

ing

of fo

ods

P2

And

erss

on, C

-G &

Lin

gner

t, H

Fä

rsk

Fors

knin

g, S

IK, G

öteb

org,

7

Febr

uary

200

7 A

tt u

ndvi

ka a

kryl

amid

när

man

bak

ar (

To

avoi

d ac

ryla

mid

e w

hen

baki

ng)

P6

J.J.

Kno

l, J.

P.H

. Lin

ssen

and

M

.A.J

.S. v

an B

oeke

l Ph

D s

tudy

trip

USA

, pre

sent

atio

ns a

t di

ffer

ent u

nive

rsit

ies

in th

e U

SA

Kin

etic

Mod

elin

g: A

Too

l to

Und

erst

and

the

Form

atio

n of

Acr

ylam

ide

(Ora

l pre

sent

atio

ns)

P7

J.J.

Kno

l, G

. Vik

lund

, J.P

.H.

Lins

sen,

I. S

jöho

lm, K

. Sko

g,

M.A

.J.S

. van

Boe

kel

CO

ST 9

27 A

ctio

n W

orks

hop,

Sof

ia,

Bul

gari

a K

inet

ic M

odel

ing:

A T

ool t

o U

nder

stan

d th

e Fo

rmat

ion

of A

cryl

amid

e in

Foo

d (O

ral p

rese

ntat

ion)

P7

Vác

laví

k L.

, Bar

táko

vá V

., H

olad

ová

K.,

Haj

šlov

á J.

H

eato

x Pr

ojec

t Mee

ting

Pra

gue,

Cze

ch

Rep

ublic

, 21.

-23.

1.20

07

Fast

and

sim

ple

met

hod

for

acry

lam

ide

anal

ysis

and

con

tent

of

acry

lam

ide

in C

zech

bre

ads

P10

of

77

Siri

Hel

land

Han

sen

and

Gun

nar

Bru

nbor

g In

tern

al s

emin

ar a

t the

Div

isio

n of

E

nvir

onm

enta

l Med

icin

e, N

orw

egia

n In

stit

ute

of P

ublic

Hea

lth

Inta

ke o

f cri

sp b

read

and

cof

fee

whi

le m

akin

g ch

ildre

n –

mut

ually

ex

clus

ive?

(in

Nor

weg

ian)

P1

6

Siri

Hel

land

Han

sen,

Min

h H

oang

, Eri

k Sø

derl

und

and

Gun

nar

Bru

nbor

g

HE

AT

OX

36

mon

th m

eeti

ng, P

ragu

e,

The

Cze

ch R

epub

lic, 2

1st-2

3rd o

f Jan

. 20

07

Gen

otox

ic e

ffec

ts o

f gly

cida

mid

e on

mou

se te

stic

ular

cel

ls

P16

J.J.

Kno

l, J.

P.H

. Lin

ssen

and

M

.A.J

.S. v

an B

oeke

l 9t

h In

tern

atio

nal S

ympo

sium

on

the

Mai

llard

Rea

ctio

n Fa

te o

f acr

ylam

ide,

mel

anoi

dins

, sug

ars,

asp

arag

ine,

org

anic

aci

ds a

nd

pH in

an

aque

ous

fruc

tose

-asp

arag

ine

reac

tion

sys

tem

at p

H 5

.5: a

ki

neti

c an

alys

is (

Subm

itte

d fo

r O

ral p

rese

ntat

ion)

P17

Oth

er w

ays o

f di

ssem

inat

ion

Ani

ka d

e M

ul, P

olly

E. B

oon,

Ja

cob

D. v

an K

lave

ren

Tra

inin

g fo

r E

FSA

and

Nat

iona

l Foo

d A

utho

riti

es

Tra

inin

g pr

ogra

m o

n pr

obab

ilist

ic e

xpos

ure

asse

ssm

ent f

or

inte

rnat

iona

l and

nat

iona

l foo

d sa

fety

aut

hori

ties

P17

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