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2 Food and Chemical Toxicology 50 (2012) S724 S735 Contents lists available at SciVerse ScienceDirect Food and Chemical Toxicology journal homepage: Application of the BRAFO tiered approach for benefit risk assessment to case studies on heat processing contaminants Katrin Schütte a, Heiner Boeing b, Andy Hart c, Walther Heeschen d, Ernst H. Reimerdes e, Dace Santare f, Kerstin Skog g, Alessandro Chiodini h, a Procter & Gamble Eurocor, Temselaan 100 Box 43, 1853 Strombeek-Bever, Belgium b Institute of Human Nutrition (DIFE), Department of Epidemiology Potsdam, Rehbrücke, Arthur-Scheunert-Allee , Nuthetal, y c The Food and Environment Research Agency (FERA), Risk Analysis Team YO41 1LZ York, UK d Federal Dairy Research Centre, Dielsweg 9, Kiel, Gemrany e FoodInfoTec (FIT), 1096 Bourg-en-Lavaux, Switzerland f Food and Veterinary Service of Latvia 30 Pelds Str., 1050 Riga, Latvia g University of Lund, Centre for Chemistry and Chemical, Engineering Box 124, Lund, Sweden h ILSI Europe, Avenue E. Mounier 83, Box 6, 1200 Brussels, Belgium article info abstract Article history: Available online 7 February 2012 Keywords: Benefit risk assessment Tiered approach Heat processing Acrylamide PAH Milk The aim of the European Funded Project BRAFO (benefit risk analysis of foods) project was to develop a framework that allows quantitative comparison of human health risks and benefits of foods based on a common scale of measurement. This publication describes the application of the BRAFO methodology to three different case studies: the formation of acrylamide in potato and cereal based products, the formation of benzo(a)pyrene through smoking and grilling of meat and fish and the heat-treatment of milk. Reference, alternative scenario and target represented the basic structure to test the tiers of the framework. Various intervention methods intended to reduce acrylamide in potato and cereal products were evaluated against the historical production methods. In conclusion the benefits of the acrylamide-reducing measures were considered prevailing. For benzo(a)pyrene, three illustrated alternative scenarios were evaluated against the most common smoking practice. The alternative scenarios were assessed as delivering benefits, introducing only minimal potential risks. Similar considerations were made for heat treatment of milk where the comparison of the microbiological effects of heat treatment, physico-chemical changes of milk constituents with positive and negative health effects was assessed. In general, based on data available, benefits of the heat treatment were outweighing any risks. Ó 2012 ILSI Europe. Published by Elsevier Ltd. All rights reserved. 1. Introduction and methodology Food matrices are highly complex systems of proteins, carbohydrates, lipids, minerals and other nutrients. Due to the components individual reactivity substantial interactions and changes occur during heat processing. That processing can induce desired and positive changes to a food product, but on the other side also Abbreviations: AA, acrylamide; BRAFO, benefit risk assessment of foods; BaP, benzo(a)pyrene; BMDL, benchmark dose (lower confidence limit); GA, glycidamide; IARC, International Agency for Research on Cancer; MOE, margin of exposure; NIEHS, National Institute of Environmental Health Sciences; PAH, polycyclic aromatic hydrocarbon; PTDI, provisional tolerable daily intake; UHT, ultra high treatment; USL, upper safe level. Corresponding author. Address: ILSI Europe a.i.s.b.l., Avenue E. Mounier 83, Box Brussels, Belgium. Tel.: +32 (0) ; fax: +32 (0) address: publications@ilsieurope.be. leads to formation of heat-formed contaminants that are potentially hazardous to human health. Hence, the risk benefit assessment methodology proposed by Hoekstra in context of the European Funded Project BRAFO (benefit risk analysis of foods) (Hoekstra et al., 2010) appears to be worthwhile to test on different heat-processing situations for different foods. Benefit risk analysis is supposed to give a clear picture of quality profiles of food systems and allow their optimization via positive balancing of the benefit risk ratio for suitable nutrition (Fig. 1). 2. Case study: Acrylamide One example of a heat-processing contaminant evaluated in this BRAFO work package is acrylamide formed in potato and /$ - see front matter Ó 2012 ILSI Europe. Published by Elsevier Ltd. All rights reserved. doi: /j.fct

3 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S735 S725 Pre-assessment and problem formulation Tier 1 Individual assessment of risks and benefits no benefit no risk Reference scenario Alternative scenario Stop: advise reference Stop: advise alternative both risks and benefits Tier 2 Qualitativeintegration of risks and benefits no clear dominance risks clearly dominates benefits benefits clearly dominates risks Stop: advise reference Stop: advise alternative Tier 3 Deterministic computation of common health metric worst/bad case analysis Sensitivity analysis Increasingly assessing more and more parameters probabilistically Tier 4 Probabilistic computation relatively small uncertainties Net benefit < 0 advise reference Net benefit > 0 advise alternative large uncertainties Health units Fig. 1. A flow chart of the BRAFO tiered approach for health risk benefit assessment of different dietary scenarios (reference and alternative). The formulation of the risk benefit question may be iteratively refined in consultation with the risk manager/policymaker as the assessment progresses, as indicated by the dashed arrows at the left side of the figure. (from Hoekstra, J., et al. BRAFO tiered approach for benefit risk assessment of foods. Food Chem. Toxicol. (2010)) cereal based products. Acrylamide has been selected as a case study because it is one of the best characterized heat-processing contaminants known in food. It is an animal carcinogen and while human epidemiological data is not fully conclusive, results from animal studies suggest this contaminant could impair health (Shipp et al., 2006; Klaunig, 2008). Since 2002, industry efforts have far advanced to identify potential mitigation options to reduce the level of acrylamide formation in the concerned foods. However, little investigations have been done so far on whether these mitigating interventions come with any undesired side effects. Other parameters of the food might be changed which might impact the benefit risk equation of consuming potato-based and cereal-based products together with a certain intake of acrylamide. This work looked at the problem of acrylamide formation in foods and potential changes in the benefit risk equation if food processing is changed such as to reduce acrylamide formation. The working group did not look at the beneficial effects of heat processing of foods generally this is dealt with in a comprehensive ILSI review on the same topic (van Boekel et al., 2010). The main purpose of this case study is to evaluate how well the BRAFO tiered methodology can be applied to the evaluation of reduced intake of heat-processing contaminants like acrylamide. Acrylamide (AA) is an industrial chemical and a food contaminant which may be formed in foods, particularly carbohydrate-rich and protein-low plant commodities, during cooking, frying, baking or roasting at temperatures of 120 C or higher. Most foods made from cereals, potatoes and also coffee, and some other foods have been shown to contain AA. The health effects of acrylamide have been extensively reviewed (Exon, 2006; Klaunig, 2008; Shipp et al., 2006; LoPachin et al., 2008; JECFA, 2010). The critical effects of AA are neurotoxicity (mostly relevant in occupational industrial exposure only) and carcinogenicity. The compound has been recognized as being genotoxic and carcinogenic in laboratory animals and is categorized as probable human carcinogen (IARC, 1995). The margin of exposure (MOE) calculated for acrylamide in food is low and this may indicate a human health concern (EFSA, 2005). Animal studies showed that AA is metabolized to glycidamide by CYP2E1. Glycidamide (GA) forms DNA-adducts, which are assumed being responsible for the carcinogenic effects. The latest bioassays conducted in rats and mice by the US National Toxicology Program (NTP) program have confirmed acrylamide as a carcinogen acting via its metabolite glycidamide and a genotoxic mechanism (NTP, 2011). In the human body, hemoglobin adducts of AA and GA have been established as biomarkers of exposure to these compounds (Dybing et al., 2005; Ogawa et al., 2006). A recent bio-monitoring survey of a representative subset of the US conducted by National Institute of Environmental Health Sciences (NIEHS) showed a large variability in AA and GA adduct levels between individuals. This variability was considered to be mostly due to different individual intakes of AA from food rather than from other exposure sources like smoke or industrial contact. Due to different consumption patterns of the high risk foods for AA in the different European countries (e.g. high coffee consumption in the Nordic countries/high bread consumption in -speaking countries/high consumption of fried potato products in the UK and Benelux) exposure may vary between a mean intake of mg/kg body weight (bw) per day and a high intake of mg/kg bw per day (JECFA, 2010). Mean acrylamide exposure in Europe is estimated to range between 0.31 and 1.1 lg/kg bw per day for adults (>18 years), between 0.43 and 1.4 lg/kg bw per day for adolescents (11 17 years), between 0.70 and 2.05 lg/kg bw per day for children (3 10 years) and between 1.2 and 2.4 lg/kg bw per day for toddlers (1 3 years) (EFSA, 2011). Formation of AA during food processing can be controlled to some extent, but it will never be possible to completely eliminate AA from our diets. All AA-reduction interventions that have been tried and are for a good part being commercially applied by industry today are summarized in a guidance document called the European Food and Drink Manufacturers Association (CIAA) Toolbox, published by the members of the (CIAA). ( brochures_form.asp?doc_id=65, last update February 2009, next update September 2011).

4 S726 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S735 Depending on the consumption pattern of AA-containing foods the outcome of the benefit risk assessment will likely vary Pre assessment and problem formulation The BRAFO methodology, describes in its first step the benefits and risks potentially involved in a particular exposure situation. In this case study, focus has been on the comparison of the situation regarding AA intake as it was prior to 2003 where little to no AAcontrol was applied (reference scenario) with an alternative scenario describing an AA intake via the same foods to which AAreduction measures have been applied in production (alternative scenarios). For a complete set of information all scenarios are listed below: is that AA is indeed a human carcinogen and that the most relevant cancer types are renal cell, endometrial and ovarian cancer based on the two epidemiological studies known thus far which have shown a positive correlation (Hogervorst et al., 2007, 2008). However, the totality of the epidemiological evidence does not allow clear-cut conclusions on these or any other cancer types yet. An epidemiology expert group concluded at the EFSA Scientific Colloquium in 2008 that the epidemiological evidence to date probably rules out a very strong effect on risk of most cancers from dietary intake, but advised that also the low relative risk ratios observed in some studies could be of importance in public health terms given the widespread exposure through the diet. The potential risks from use of different acrylamide reduction tools are: Reference scenario: Alternative scenarios: Cooking practices before AA-mitigation measures were introduced (AA exposure estimated: mg/kg bw/ day) AA-mitigation methods (AA exposure estimated: 30% reduction = mg/kg bw/day) Lower frying temp NaCl addition Ca-salt addition Na-bicarbonate use instead of ammoniumbicarbonate as raising agent ph-reduction Use of asparaginase Increased Ca-intake (Ca-salt addition in potato products) Increased Na-intake (raising agent replacement in bakery ware, NaCl addition in potato products) Increased 3-MCPD intake (ph reduction in bakery ware) Increased fat intake (lowered frying temperature in potato products) Some loss of asparagine (use of enzyme asparaginase in potato and cereal products) The different alternative scenarios all constitute food preparation methods that lead to a lower AA content of the respective foods. However, other factors of the diet are altered to a varying extent due to the AA mitigation effort and these side effects are being looked at with the BRAFO methodology to analyse if they represent a risk or a benefit in comparison to the benefit of reduced AA uptake. The alternative scenarios do not cover all possible mitigation scenarios, but the main ones. Acrylamide exposure occurs mainly through the diet and the margin of exposure (MOE) is smaller than desirable. Comparing typical intake levels for the average adult from all dietary sources (prior to mitigation options being applied) reported by competent international bodies of lg/day or 1 lg/kg bw/day with the Benchmark Dose (BMDL) 10 for mammary tumours in rats, the MOE values are 310 for mean exposure and 78 for high exposure, much smaller than 10,000 which has been described as of no human health concern and risk management actions. Compared with the BMDL10 for Harderian gland tumours in mice, the MOE values are 180 and 45 for mean and high exposures, respectively (JECFA, 2010). For details on the general approach how these MOE values were derived also in the JECFA assessment please see Benford et al. (2010) and Bolger et al. (2010). While there is some question mark around the relevance of Harderian gland tumours to humans (who do not have this organ), the occurrence of Harderian gland tumours in mice are almost exclusively associated with genotoxic carcinogens that are active at multiple sites. In addition to the limited relevance of Harderian gland tumours for humans, also the considerably higher metabolic activation rate of the mouse, as compared to humans, deserves mentioning (Sumner et al., 1999). Hence regulators, risk assessors and industry view strategies that reduce the AA content in foods as a health benefit. However, effects brought about by these AA-reduction interventions could lead to undesired effects or potential health risks, which are attempted to be evaluated here. The postulated benefit of acrylamide reduction in foods is: Reduction of acrylamide exposure = human health benefit due to reduced potential cancer risk. This assumption makes a simplification for the purpose of testing this BRAFO benefit risk assessment methodology. The assumption Lower frying temperature (which mostly means longer frying time) have been shown to limit AA formation but also to lead to an increased fat content of chips and fries or other fried potato and/or cereal based products. ph reduction in bread and bakery ware has been shown to limit AA formation but can bring about two side effects: the increase in Na-content of the baked good if ph changes is achieved through exchange of ammonium bicarbonate with sodium bicarbonate and/or the formation of another process contaminant, 3-MCPD, which is described to form in doughs at a higher rate if the ph is lowered. 3-MCPD is described as a non-genotoxic carcinogen (EC-SCF, 2001). The addition of salt (NaCl) has been shown to limit AA formation in bread or other dough-based products. However, this reduction tool would lead to an increased Na-intake. Also the addition of Calcium salts was shown effective in reducing AA levels in snacks products. Consequently, the Ca-intake might be increased. The enzyme asparaginase has been described as a tool to eliminate the amino acid asparagine, one of the starting materials in AA formation. The effect of this reaction is a decreased asparagine-content of the foods where the enzyme was used during production Acrylamide intake considerations Mean and high acrylamide intake values are taken from the assessments of JECFA (2010) which had also been considered relevant by EFSA (2008a,b). Monitoring data collected by the EU member states and summarized by EFSA show that while not for all product types mitigation tools were developed successfully, significant reductions have been achieved in some critical product groups:

5 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S735 S727 French fries and fried potato products for home cooking, soft bread, bread not specified, infant biscuit, biscuit not specified, muesli and porridge and other products not specified showed statistically significantly lower levels of acrylamide in 2008 data compared to 2007 data (Table 1). No statistical difference was shown for the remaining (50%) of the food sub-groups in the mean acrylamide content between the sampling years 2007 and Based on the above, a maximum intake reduction of 30% for acrylamide through the total diet is assumed for the alternative scenario in this evaluation BRAFO tier 1 level (a) Acrylamide reduction through single or combined mitigation tools/reduction of potential cancer risk All alternative scenarios describe a technological intervention to reduce AA levels of certain foods. Based on currently known data (which reports successful AA reduction in some foods but less so in others) the overall dietary AA intake could be decreased by an estimated 30%, potentially also more in future. Although difficult to quantify, the reduced exposure to AA must be considered as a decrease in potential incidence of cancers, hence an overall benefit at tier 1 (Hoekstra et al., 2010). (b) Increased Calcium intake and adverse effects (e.g. kidney effects/hypercalciuria) The addition of certain divalent metal ions like calcium has been reported (CIAA toolbox 2009) to reduce acrylamide formation in several food categories. It is assumed that these ions interact with the asparagine in the foods and make it less available for the reaction with reducing sugars to form acrylamide. Addition of up to 0.3% CaCl 2 or other Ca-salt to a dough-based product (e.g. bread, bakery products, formed potato crisps, other dough-based snacks or pommes croquettes) can reduce acrylamide levels by ca. 30%, with limited negative impact on product characteristics and organoleptic properties. In dough-based snacks up to 1% CaCl 2 can be used which can reduce AA by 20 80% (CIAA toolbox 2009). The 0.3% addition is equivalent to an addition of ca mg Ca per kg of product. When other calcium salts are used (e.g. calcium lactate) the amount of added calcium is reduced, in the case of calcium lactate to ca. 600 mg Ca/kg of product, with a similar acrylamide reduction seen. The levels of addition considered here are comparable to the calcium levels naturally found in a wide range of foods: the most important dietary calcium source, milk, contains typically 1200 mg Ca/L. The following evaluation assesses the potential increase in exposure to Calcium under estimated maximum addition and intake conditions. Exposure assessment increased Ca-intake: Consumption of potato Median: 116 g/day products (boys years, DGE, 2008) Mean + 2 SD: 302 g/day Median calcium intake 1518 mg/day (baseline) (Boys years, DGE, 2008) (P10/P90: mg/ day) Added calcium in those 1100 mg/kg products All consumption figures are taken from the Nutrition Report (Max Rubner Institute, 2008), which provides the most recent food consumption data currently available; the data are consistent with consumption data published earlier for other countries. The group of year old boys is the with the highest consumption of fried potato products; based on the Table 1 Adaptation from scientific report of EFSA: results on acrylamide levels in food from monitoring year 2008 (EFSA 2010). Mean values in lg/kg % Reduction Crackers Infant biscuits Soft bread Bread not specified French fries Home-cook potato product (oven) Home-cook potato product (deep-fry) published data, it is assumed that using their intake figures for potato products provides for a worst-case exposure assessment in this alternative scenario g 1100 mg/1000 g = mg/day g 1100 mg/1000 g = mg/day Added exposure through consumption of potato products with 0.3% CaCl 2 (1100 mg/kg): - Mean potato products consumer: 128 mg/day = 1.8 mg/kg bw/ day - High potato products consumer: 332 mg/day = 4.7 mg/kg bw/ day The addition of calcium to the products would lead to an additional intake of 128 mg of Ca/day; for the high end consumption the intake would be 332 mg additional Ca/day. The mean calcium intake reported for this group is 1518 mg/ day; the figures for the 10th and 90th percentile are 871 and 2603 mg/day. The Upper Safe Level for calcium has been proposed at 2500 mg/day. It is recommended in the general medicinal literature not to exceed this value including foods, drinks and supplements. This Upper Safe Level (USL) for daily calcium intake in adults is the highest level that likely will not pose risks of unwanted side effects in the general. This means that even for individuals with a high end intake of Ca from treated products, the additional 332 mg would raise the overall intake for this group only to 1850 mg/day, a level still below the USL. The combination of both high end consumption of treated potato products and of high end background intake of calcium would be inappropriate, since boys eating large amounts of fried potato products are usually low consumers of milk/dairy products and calcium-rich mineral waters, which today provide more than 77% of the dietary calcium for teenage boys (Mensink et al., 2007). Thus, even for the group with the highest dietary calcium intake, there is no indication that the USL for calcium would be exceeded due to the addition of calcium salts to the selected food groups Calcium intake and kidney stones First adverse effects like hypercalciuria and stone formation are observed in healthy people only at much higher intake levels (>5000 mg/day) and usually only when some medical conditions occur, which are very rare for young people (Nordin, 1988). It should be noted that for elderly men, where side effects are more likely to occur, both the actual intakes (median: 634 mg/day) and the likelihood of high intake from treated foods are much lower than those for teenage boys (Max Rubner Institute, 2008). The same applies for elderly women. At intakes not exceeding the USL of 2500 mg/day, no risk is to be expected. The largest prospective epidemiological study published on calcium and kidney stones concluded that high calcium

6 S728 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S735 intake is associated with a decreased risk of symptomatic kidney stones (Curhan et al., 1993). Perhaps just as importantly, the study, conducted among over 45,000 men, found that those individuals who consumed less than 850 mg of Ca/day were at an increased risk for a higher incidence of kidney stones. The authors concluded that calcium might actually have a protective effect by binding to oxalate in the gut and preventing its absorption in a form that leads to kidney stones. This conclusion was supported by a subsequent study on long-term calcium supplementation in pre-menopausal women, which found no increase in stone formation (Sakhaee et al., 1994). Calcium supplementation lowered both urinary oxalate and urinary phosphorous (also thought to contribute to the formation of stones) by binding both agents in the intestine. It seems demonstrated that the addition of calcium for the benefit of AA mitigation is not likely to lead to exceeding the Upper Safe Level, even under worst case assumptions. Thus, the potential health impact of this alternative scenario b is assessed as none and evaluation of this scenario hence stopped at tier 1. (c) Increased calcium intake and bone health For evaluation of the effects on bone health, the same exposure assumptions as in scenario (b) are being used as maximum intake changes. The consideration of calcium intake for bone health is most relevant in women where osteoporosis is more frequent than in men. The background calcium intake in teenage girls (12 18 years) is not favorable. The median intake is around the DACH-recommendations (DGE, 2000), thus nearly half the girls (similar to the adult women) would actually benefit from an increased calcium intake. The intake for the 10th percentile was as low as 665 mg/day. An appropriate calcium intake would decrease their risk to develop osteoporosis later in life: in a 14-year prospective study by Holbrook et al. (1988), below the group cut-off intake of 765 mg Ca/ day the risk for osteoporotic fractures had increased by 60%. Other more recent studies have confirmed this benefit although the risk reduction has been less pronounced. It is important to note that it is calcium intake over the whole lifetime, especially also during young age, which determines the risk for osteoporosis later in life. The actual dietary calcium intake is even lower in adult and especially in elderly women (65 95 years) with a median intake at only around 60% of the Division of Adult and Community Health (DACH) recommendations (DGE, 2008). For them, next to potato products, bread and other bakery products would be the main sources of acrylamide, thus categories where addition of calcium could also be a potential mitigation step. If one includes the potential benefit of an increased calcium intake on bone health, i.e. bone formation at young age and reduced bone loss later in life, there seems to be a potentially significant reduction in the risk of osteoporotic fractures. Based on the Ca-intake data available for women and the positive effects described on bone health through an adequate or increased Ca-intake, the overall health impact of this alternative scenario (c) is judged as beneficial. (d) NaCl addition for AA mitigation/increased Na-intake Two mitigation options achieving a reduction of AA levels associated with an increase in sodium content of the respective products have been identified, i.e. a change in raising agents from ammonium to sodium bicarbonate in some fine bakery products, and the addition of table salt to the dough in a much wider range of foods, including bread and dough based potato products. While considered an effective tool initially, the addition of salt does not seem to be widely applied in industry today, however is considered in this risk benefit assessment exercise as an illustration of the methodology. The replacement of the raising agent ammonium bicarbonate by sodium bicarbonate in ginger bread and certain biscuits has been recently assessed in detail for its impact on dietary sodium intake and related risks (Seal et al., 2008). As expected, the increase in sodium intake turned out to be limited due to the low consumption figures for the small number of foods involved. Only 1.3% of the would be expected to shift from a Na intake below 40 mg/kg/day to above this figure, and the net increase through AA mitigation in fine bakery ware remains small Exposure estimation for replacing NH 4 CO 3 with NaHCO 3 In biscuits and gingerbread, the replacement of NH4 with Na leads to an increase in sodium content of the products, from 140 mg/kg product to 500 mg/kg product. Intake data cookies: National Food Consumption Survey 2008 (Max Rubner institute, 2008) Men 46 g/day, women 33 g/day, average both sexes = 40 g/day (likely a high estimate, as intake is for bakery wares in total) Additional Na-intake through this reference scenario: 40 g/day at Na-level 140 mg/kg = 5.6 mg/day 40 g/day at Na-level 500 mg/kg = 20 mg/day Additional Na-intake of 15 mg/day through replacement of the raising agent. Starting from the findings by Seal et al. (2008), the direct addition of sodium as sodium chloride to a wider range of foods to reduce acrylamide may lead to a significantly higher net increase in Na intake and thus warrants a more detailed evaluation. Kolek et al. observed in simple food model systems that the addition of NaCl favours the further polymerization of acrylamide during heating and can hence lead to a decrease in AA content (Kolek and Simon, 2006, 2007). Early observations had shown that the addition of an additional ca. 1% of NaCl to dough based products like bread or some potato products or the soaking of French fries in salt solution prior to frying can be associated with a reduction of acrylamide formation by 20 30% (Seal et al. 2008). Especially in the case of bread, such an additional intake in salt levels would run counter to current EU attempts to limit the total salt addition as a step against the rising incidence of hypertension in Europe. The ingoing assumption, based on the technical information available from industry, is that an addition of 1% salt to dough based products, especially bread and some other non-sweet bakery products can reduce acrylamide levels by 30%. The following assumptions are used: Intake data based on latest figures (DGE, 2008). Actual consumption of bread in young men: Mean: 234 g High end (Mean + 2 SD): 494 g 1% Extra salt = 1 g extra salt/100 g bread = 0.4 g extra sodium/100 g bread (salt = 40% sodium by weight) 0.4 g 2.34 = g extra sodium (mean consumer) 0.4 g 4.94 = g extra sodium (high consumer) Current background Na intake in young men: Mean: 4.1 g/day 10th centile: 2.5 g/day 90th centile: 6.2 g/day The use of consumption figures appears right not only because they present a recent set of data (published in December 2008) but consumption of bread and sodium intake seem to be at the high end of Europe. Figures for Denmark are comparable to y, data for The Netherlands (Zo eet Nederland, 1998), Austria (Elmadfa, 1998) and UK show lower consumption. For

7 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S735 S729 countries like Poland or Italy, where also high bread consumption has been reported by Elmadfa (1998), no specific Na-content data are currently available. Based on these assumptions, we find an incremental Na intake from mean bread consumption of 0.94 g/day and even 1.98 g/day for individuals with high-end bread consumption. It can be assumed that an increase in Na intake is closely mirrored in urinary Na excretion. In this context, a meta-analysis of 40 intervention studies on sodium intake and blood pressure (Geleijnse et al., 2003) can be useful. In this study, a significant reduction of systolic blood pressure was observed between a decrease in urinary Na excretion above and below 77 mmol (1.8 g) sodium per day. Based on the figures presented above on incremental Na intake from the added salt, one can assume that the intake/excretion of the average of young males and likely parts of other subs would be pushed above the excretion limit determined in this review. For high end bread consumption, the incremental Na intake of 1.98 g/day alone would be comparable to the cut-off level in the Geleijnse-Study. It is important to note that already during childhood and adolescence an increased salt/sodium intake is associated with an increased blood pressure (He et al., 2008). In addition, it is being criticized that early consumption of high salt foods conditions people towards an overall high salt consumption over their lifetime (DGE, 2008). The review by Geleijnse et al. (2003) also indicates that with increasing age the sensitivity of blood pressure to increased Na intake increases. Based on data from Max Rubner Institute 2008 (adjusted from their energy intake), elderly men (mean age 70 years) in y are consuming 2.7 g of Na per day. Their bread consumption is comparable to the mean of young adults, since the overall lower energy intake is not reflected in a reduction of bread consumption: bread remains a key part of their traditional diet. With this background, the use of salt/sodium to reduce acrylamide formation would lead to an increase of their daily Na intake of nearly 1 g, i.e. by about one third of their current average. The data suggest that such a significant increase in the salt content of bread must be assumed to have a noticeable undesired effect on the blood pressure of consumers in general. Clearly, this probable influence on blood pressure changes must be considered together with the so far not fully conclusive evidence of a causative effect of increased blood pressure on cardiovascular diseases like stroke or infarction. Overall, for this alternative scenario an increased incidence of the health impact has to be assumed which leads to describing the health impact at tier 1 level as adverse. (e) Lowered dough ph/increased potential for 3-MCPD formation Increased 3-monochloropropane-1,2-diol (3-MCPD) formation at lowered ph values in bread and bakery ware was shown (Seal et al., 2008). 3-MCPD was recently reviewed by the International Agency For Research on Cancer (IARC) (Grosse et al., 2011). In rats, it produced benign renal tumours in both sexes and Leydig-cell and mammary tumours in males. 3-MCPD is genotoxic in vitro, but there is no evidence of genotoxicity in vivo. The potential health effect of increased intake of an animal carcinogen is considered relevant. However, 3-MCPD is currently regulated by a maximum level in products where its formation occurs the most and a provisional maximum tolerable intake (PMTDI) (2 lg/kg bw/day) has been published (WHO, 2007). For this assessment, the group concluded that a health effect should be excluded unless the exposure would be estimated to exceed the PMTDI. Data on 3-MCPD formation in bread and bakery dough at lowered ph is still limited. Based on the analytical values available, the following exposure estimation was conducted: Exposure: 3-MCPD levels in continental breads (normal ph): [lg/kg] 400% increase observed as a maximum from addition of acids: 50 4 = 200 = lg/kg bread Bread consumption (DGE, 2008): Mean: High end (Mean + 2 SD): 234 g 200 lg/kg/1000 g = 47 lg/day 494 g 200 lg/kg/1000 g = 98 lg/day Exposure at 400% increased 3-MCPD production: Mean bread consumer: 47 lg/d = 0.7 lg/kg bw/ day High bread consumer: 98 lg/d = 1.4 lg/kg bw/ day 234 g/day 494 g/day The exposure to 3-MCPD through increased formation in low ph dough is even under worst-case assumption (400% 3-MCPD formation increase and high bread consumption) still below the PTDI. Consequently, the health impact of this alternative scenario d is assessed as none and evaluation of this scenario hence stopped at tier 1. The authors would like to point out that only 3-MCPD exposure from bread consumption was considered for this assessment, as these were the only data available of a scenario where acrylamide mitigation has impacted 3-MCPD levels in foods. There are not sufficient data available currently to allow conclusions on whether acrylamide mitigation tools applied to other food categories like e.g. cereals or coffee could also increase the uptake of 3-MCPD in the. (f) Lower frying temperature/increased fat content Company-owned unpublished data from various snacks and French fries producers from the years show that a frying temperature reduction from e.g. 200 to 175 C leads to 30% decrease in AA and can lead to a 10% increase of fat content of the fried product. A potential health effect of increased fat uptake would be an increased risk of hyperlipoproteinaemia and cardiovascular diseases. Today, based on increased industry learning, these estimated 10% increased fat content are not considered to be occurring in practice to the total as products that are commercially fried can undergo other formulation adaptations to avoid such an increased fat content. The increased fat uptake through reduced frying temperature would then remain relevant only for homecooked freshly fried goods, which is likely to be a negligible change in overall fat uptake through the diet. The health impact of this alternative scenario c is assessed as none and evaluation of this scenario hence stopped at tier 1. (g) Enzyme Asparaginase/loss of asparagine For completeness it is mentioned here that also the use of asparaginase is a possible AA reduction tool, effective in some food products. The enzyme is commercially available as product from two different production organisms, and both have received safety approval (US FDA GRAS status for their intended use). The enzyme reduces asparagine (transforming it to aspartate) and hence will reduce the level of asparagine in the foods on which the enzyme is used during production. However, given that asparagine is a non-essential amino acid and given the positive safety evaluation of the enzyme for use in food production there is no relevant health impact to be expected. This alternative scenario is stopped for evaluation at tier 1.

8 S730 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S BRAFO tier 2 level In tier 2, both benefits and risks should be characterized based on the severity of the effects and on the potential number of individuals affected (Hoekstra et al., 2010). For the AA case study, health effects of high severity included cancer risk (reduced here) and hypertension and associated CVDs (increased risk), while a medium to low rank was attributed to the effect improved bone health/reduced fractures (Table 3). However, the authors found the precise health significance of an acrylamide reduction of 30% in some food products difficult to assess. The same holds true for the actual effect of the increased Caintake for bone health. Dose response curves for dietary acrylamide exposure and adverse effects in humans are not available. Some data exist in experimental animals (at exposures different to those through the human diet). However, no standard methodology is currently developed to extrapolate human dose response from animal data in the context of the BRAFO benefit risk assessment methodology Conclusion In tier 1, as the overall evaluation in Table 2 outlines, a resolution of the benefit risk assessment for acrylamide and mitigation efforts could not be found at that level as a potential beneficial and a potential adverse effect contrast each other. However, some risks or benefits could be ruled out from occurring based on available realistic data; based on low quality of evidence or small change in incidence based on practical industry experience: increased fat uptake, increased 3-MCPD intake, Ca-intake above USL, and enzyme influence on asparagine intake have been ruled out. In tier 2, as the overall evaluation in Table 3 outlines, a resolution of the benefit risk assessment for acrylamide and mitigation efforts could not be found at tier 2 yet either, as a potential decrease in mortality (through reduced AA exposure, and through additional Ca-intake considered beneficial for bone health) is to be evaluated against a potential increase in mortality from another effect (potentially increased Na-uptake). Hence the next steps according to the methodology would be to estimate the corresponding DALY changes of both effects. This could theoretically be tried out using the QALIBRA tool, however, the authors concluded that not sufficient data of satisfactory quality necessary to undertake this modelling were available. Overall, it can be concluded though that the beneficial effect of reducing acrylamide in processed foods through various intervention methods in the food production is desirable. These acrylamide-reducing actions should hence be applied as long as the adverse side effects are recognized and minimized to the extent possible. 3. Case study: Benzo(a)pyrene 3.1. Background Benzo(a)pyrene (BaP) is a chemical contaminant found in food. BaP belongs to the group of polycyclic aromatic hydrocarbons (PAHs), which generally occur in complex mixtures. The contamination of food with PAHs can occur both by environmental pathway and during heat treatment applications in food processing, e.g. roasting, grilling, smoking, drying. PAHs are primarily formed by incomplete combustion or pyrolysis of organic matter, during various heat processing. There are several food categories that can be affected by PAHs, such as cereals, vegetable oils, coffee, home-cooked foods, usually when smoking, heating and drying processes are involved, or seafood from polluted waters. Home cooking, such as grilling, roasting and smoking, particularly charcoal grilled/barbecued foods, can lead to high concentrations of PAHs. One the most popular and oldest food processing technology relevant to the issue is smoking. The technology is widely used not only for special organoleptic properties of smoked products, but also for the inactivating effect of smoke and heat on enzymes and microorganisms (Stolyhwo and Sikorski, 2005). It has to be stated that, the toxicity of PAH is not well characterized. The toxic equivalency factors (TEF) approach to the risk characterization for PAHs in food is not considered to be scientifically valid because of the lack of data from oral carcinogenicity studies on different PAHs, their different modes of action and the evidence of poor predictability of the carcinogenic potency of PAH mixtures based on the currently proposed TEF values (EFSA, 2008a). PAHs differ in carcinogenic potency or may function in living organisms as synergists increasing carcinogenic activity of other PAHs. The health effects of PAHs have been extensively reviewed (EFSA, 2007, 2008a). In the EFSA Scientific Opinion on Contaminants in the Food Chain (EFSA, 2008a) it is also concluded that BaP may be used as a marker of occurrence and effect of the carcinogenic PAHs in food. However, it is stressed the fact that data collection on the whole PAH profile should continue in order to be able to evaluate the contamination of food commodities and any future change in the PAH profile Exposure to BaP and PAHs via the diet According to EFSA (2007, 2008b)) the median dietary exposure across European countries, calculated both for mean and high dietary consumers and varied between 235 ng/day (3.9 ng/kg bw per day) and 389 ng/day (6.5 ng/kg bw per day) respectively for BaP alone, 641 ng/day (10.7 ng/kg bw per day) and 1077 ng/day (18.0 ng/kg bw per day) respectively for PAH2 (BaP and chrysene), 1168 ng/day (19.5 ng/kg bw per day) and 2068 ng/day (34.5 ng/kg bw per day) respectively for PAH4 (BaP, chrysene, benzo[a]anthracene & benzo[b]fluoranthene) and 1729 ng/day (28.8 ng/kg bw per day) and 3078 ng/day (51.3 ng/kg bw per day) respectively for PAH8. The two highest contributors to the dietary exposure were cereals and cereal products, and sea food and sea food products (EFSA, 2007, 2008a). The indicated margins of exposure (MOE) for average consumers were 17,900 for BaP, 15,900 for PAH2, 17,500 for PAH4 and 17,000 for PAH8. For high level consumers, the respective MOEs were 10,800, 9500, 9900 and These MOEs indicate a low concern for consumer health at the average estimated dietary exposures. This applies to the full range of estimates of average exposures across EU Member States ( ng/kg bw per day, MOEs: 16,300 22,600 for benzo[a]pyrene alone and ng/kg bw per day, MOEs: 13,800 20,800 for PAH8). However, for high level consumers the MOEs are close to or less than 10,000, which as proposed by the EFSA Scientific Committee indicates a potential concern for consumer health and a possible need for risk management action (EFSA, 2008a,b) Pre-assessment and problem formulation BaP is a genotoxic carcinogen and human exposure to this compound via the diet should be kept to as low as can be reasonably achieved (Scientific Committee of Foods, 2002). Whilst an MOE approach indicates that for average consumers exposure is of low concern, for high consumers the MOE is not sufficient. In formulating the reference scenario consideration of BaP present in food due to smoking and grilling of fish and meat is considered. Potential environmental contamination is excluded.

9 K. Schütte et al. / Food and Chemical Toxicology 50 (2012) S724 S735 S731 Table 2 Case study on acrylamide benefit risk assessment for a change from the reference scenario to different alternative scenarios at tier 1. References Magnitude of the effect Population affected Health impact (benefit/ adverse/none) Health effect Change Quality of evidence based on WHO criteria B CIAA (2009), Exon (2006), Hogervorst (2007), Hogervorst (2008), Hubert et al. (2009); Klaunig (2008), IARC (1995) Medium Decrease Total (mainly 50 + years) Acrylamide reduction by 30 50% Tier 1 Reduced probable cancer risks (renal, ovarian, endometrial) Total N DACH (2000), Curhan et al. (1993), Sakhaee et al. (1994) Kidney issues Increased Calcium intake Low Negligible (intake low relative to background) Total B Holbrook et al. (1988), Kersting et al. (2004) Increased Calcium intake High Low (intake low relative to background) Improved bone health/ reduced fractures A DGE (2008), Geleijnse (2003), He et al. (1999), He et al. (2008), Seal et al. (2008) Increased Sodium intake Medium High Increase Total (mainly 50 + years) Increased risk of hypertension and associated CVDs Total N Hamlet and Sadd (2005), Seal et al. (2008) Increased 3-MCPD intake Medium (animal data only) Negligible (increase will not exceed current TDI) Increased probable cancer risk Total N Internal industry data Increased Fat intake High Negligible (fat increase can be avoided) Increased risk of Hyperlipoproteinaemia & CVDs Loss of asparagine N.A. Negligible Total N None (non-essential amino acid) Overall change? In order to reduce exposure to BaP from smoking or grilling of foodstuffs several alternative approaches could be used. These are presented below, taking into consideration the European as target. Reference scenario: Alternative scenarios: The potential benefits are: Smoking and grilling of fish and meat as per current practice in home, which may give rise to product contamination with PAHs Addition of artificial smoke flavours (from EFSA positive list) instead of smoking Industrial controlled smoking Consumer advice: to discourage homesmoking, to recommend using of aluminium foil for grilling, or vertical grilling The reduction of BaP and PAHs uptake is stressed as single, most important benefit due to potential impact on human health from a potential reduction in cancer risk. The potential risks are: As the alternative scenarios proposed lead to slightly changed food processing methods with the aim to achieve reduction of PAHs in the food matrices, other possible effects relevant to particular methods, substances or process may be introduced, and appear as benefit risk neutral or may raise cause to another risk or an additional benefit. For example incomplete cooking due to reducing grilling time (alternative scenario 3) BRAFO tier 1 level Alternative scenario 1: addition of artificial smoke flavours instead of smoking. Smoked fish and meat forms a significant part of the human diet, important because of their specific, desirable organoleptic profile, and the high nutritional value and abundance, in fatty species, of lipids rich in unsaturated fatty acids. One alternative food processing method to avoid formation of PAHs completely is the heat processing of meat or fish, avoiding any smoking or grilling. In order to give products the expected taste profile, smoke flavours would be added. Smoke flavours which will in future be allowed for use, based on the new FIAP regulation have all undergone a recent evaluation by EFSA and are considered safe for consumption at a defined level (EFSA positive list of smoke flavours). Based on the fact that smoke flavours have been evaluated and are considered as safe this alternative scenario of increasing their intake will not deliver any potential benefit or risk Alternative scenario 2: industrial controlled smoking. Smoking is an ancient and popular food heat processing method traditionally used by industry and in local communities. Formation of PAHs depends on numerous processing parameters like, time, temperature, source of heat, type of heat treatment and technological solutions used. Next to the very traditional, uncontrolled processing methods home grilling or home smoking, the fish and meat industry has established very well controlled smoking processes which guarantee an adequate level of preservation of the food but at the same time limit the formation of PAHs. Maximum tolerated levels of benzo[a]pyrene have been set in Regulation (EC) 1881/2006 and industrially controlled processes are supposed to monitor and respect these. We hence assume that exposure to the contaminants is reduced in this alternative scenario and are not aware of any data suggesting that other risks (e.g. through other heat-formed