Exploring Maillard reaction markers and melanoidins to investigate toxicological and antioxidant profiles of optimized expanded snacks from corn/common bean mixtures.

BACKGROUND: Extrusion cooking of cereal-legume flour mixture is an innovative strategy to introduce nutrient-enriched ready-to-eat snacks to the market. However, this thermal process triggers the formation of compounds that could impact safety aspects of these products. Maillard reaction markers and the end products known as melanoidins were evaluated to assess the toxicological and bioactive profiles of extruded snacks from corn-plus-common-bean-flour combinations. Different molecular weight fractions were isolated and purified to analyze their antioxidant activity and to investigate the role of melanoidins. RESULTS: The snack formulated with an 84:16 ratio of corn:common bean flours exhibited an enhanced toxicological profile. It displayed the lowest levels of acrylamide and furanic compounds, along with reduced blockage of lysine residues in the protein. Extrusion increased the antioxidant activity of uncooked flours (30 to 64%) and total phenolic compounds (26 to 50%), and decreased the available lysine (-72.7 to -79.5%). During the fractionation process, it was established that compounds within the range of 3-10 kDa made the greatest contribution to antioxidant activity. The fraction greater than 10 kDa, which included melanoidins, displayed 7 to 33% lower antioxidant activity. The purification of the fraction greater than 10 kDa revealed that pure melanoidins represented approximately one-third of the antioxidant activity in that fraction. Non-covalent adducts linked to the melanoidin core therefore had a relevant role in the antioxidant action of formulated snacks. CONCLUSION: This investigation illustrates the importance of considering both potential risks and associated benefits of compounds formed during the Maillard reaction while developing new extruded snacks. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Chickpea Seed Flours Improve the Nutritional and the Antioxidant Profiles of Traditional Shortbread Biscuits: Effects of In Vitro Gastrointestinal Digestion.

Functional gluten-free biscuits enriched with commercial and landrace non-commercial chickpea flours were designed and compared with a traditional shortbread biscuit. They were analyzed in sensory attributes, amino acid profile, and antioxidant properties. Subsequently, the biscuits were digested in vitro to evaluate protein hydrolysis, amino acid bioaccessibility, phenolic compounds release, and antioxidant markers. The presence of chickpea flours provided golden color and heightened biscuit hardness and fracturability (especially in non-commercial), increasing crispness and reducing brittleness. The protein hydrolysis was similar among samples (≈15%), except for one of the non-commercial (≈20%). Amino acids such as arginine, phenylalanine, leucine, tyrosine, and lysine exhibited the highest bioaccessibilities. Incorporating chickpea flour improved the antioxidant activity and polyphenol content in undigested samples and bioaccesible fractions, with higher levels of p-coumaric and ferulic acids after digestion, regardless of the chickpea seed. Non-commercial flours increased the presence of resveratrol and/or catechin in the bioaccessible fraction. Antioxidant action assessed in the Caco-2 cell line showed that the protective effect against reactive oxygen species (ROS) generation did not always correlate with the in vitro antioxidant capacity. Our data support that the inclusion of chickpea flours in the formulation of functional biscuits provides the consumer with products of added nutritional value with attractive organoleptic features.

Assessment of dietary acrylamide exposure in children attending Spanish school canteens using the duplicate diet method.

Acrylamide is a carcinogenic chemical contaminant formed in heat-treated foods. In this study, a duplicate diet method was used to evaluate the acrylamide content of foods/meals served at breakfast and lunch in two Spanish school canteens. The dietary acrylamide intake in students was estimated within lower bound (LB) and upper bound (UB) scenarios. Biscuits exhibited the highest acrylamide values, exceeding the benchmark level established by the European Regulation 2017/2158 (350 µg/kg), followed by main courses such as stews, side dishes and bread. In the LB scenario, breakfasts accounted for the major contributors to the daily acrylamide intake (73.3%). However, lunches were the main responsible in the UB scenario (65.4%). Acrylamide exposure was estimated for three age ranges: 3-5y pre-school children (0.59 and 0.92 µg/kg body weight/day), 6-9y children (0.43 and 0.67 µg/kg body weight/day) and 10-12y early adolescents (0.28 and 0.44 µg/kg body weight/day). Margins of exposure for neoplastic effects ranged between 144 and 1026, which is below the reference of 10,000, indicating a health concern. The findings reveal that any diet, even one that is balanced, varied and contains foods low in acrylamide, involves an additive exposure to the contaminant that should be considered when conducting acrylamide exposure risk assessments.

An Updated Review of Soy-Derived Beverages: Nutrition, Processing, and Bioactivity.

The global market for plant-based drinks is experiencing rapid growth driven by consumer demand for more sustainable diets, including vegetarian and vegan options. Soy beverages in particular are gaining popularity among individuals with lactose intolerance and milk protein allergies. They are considered an excellent source of high-quality protein, vitamin B, unsaturated fatty acids, and beneficial phytochemicals such as phytosterols, soy lecithins, and isoflavones. This review presents a comprehensive market survey of fifty-two soy beverages available in Spain and other European countries. The predominant category among those evaluated was calcium and vitamin-fortified drinks, accounting for 60% of the market. This reflects the need to address the nutritional gap compared to cow's milk and meet essential dietary requirements. The review covers the technological aspects of industrial soy milk production, including both traditional methods and innovative processing techniques. Additionally, it analyzes multiple studies and meta-analyses, presenting compelling evidence for the positive effects of soy beverages on various aspects of health. The review specifically examines the contributions of different components found in soy beverages, such as isoflavones, proteins, fiber, and oligosaccharides. Moreover, it explores controversial aspects of soy consumption, including its potential implications for growth, puberty, fertility, feminization, and the thyroid gland.

Dietary Exposure to Acrylamide in Spanish University Canteens by the Duplicate Diet Method.

During the university period, many students adopt new dietary patterns, sometimes including the excessive consumption of highly processed foods, which can expose them to process contaminants such as acrylamide. This research aimed to evaluate the dietary exposure to acrylamide of Spanish university students in their campus canteens using the duplicate diet method, and to estimate the associated health risks based on their food consumption habits. Apart from potato-based foods, the analysed food/meals contained non-detectable or low levels of acrylamide (<63 µg/kg). Two exposure scenarios were considered, depending on whether students selected salad (lowest exposure) or processed potatoes (highest exposure) as side dishes. The frequent consumption of processed potatoes could increase acrylamide intake from 6.5 to 26.4 µg/day. Due to their lower body weight, women had a higher risk of acrylamide exposure in both scenarios. The margin of exposure (MOE) values for neoplastic effects indicated high levels of health concern, even only considering the main meal of the day. Furthermore, the risk of acrylamide exposure could increase by more than four times depending on the side dish selected by the students. This research highlights the need to promote healthy dietary habits among young people, encouraging the selection of safer food options in terms of food processing contaminants.

Acrylamide Exposure from Common Culinary Preparations in Spain, in Household, Catering and Industrial Settings.

In 2019, the European Commission recommended monitoring the presence of acrylamide in certain foods not included in Regulation 2158/2017, to consider other sources of exposure to the contaminant. In the present study, eleven groups of processed foods commonly consumed in Spain were classified, according to their food matrix, into potato-based food, cereal-based food and food based on cereal mixed with meat, fish or vegetables. Samples were collected from three different settings: household, catering services and industrial origin, to evaluate the influence of the food preparation site on acrylamide formation. The highest concentrations of acrylamide were observed in chips (French fries), especially those prepared at home. Although at lower levels, all the other foods also contained significant concentrations of acrylamide, confirming the need to control its content in foods not included in the EU regulation. Industrially processed foods made a lower contribution to acrylamide exposure, probably due to the more stringent controls exercised on culinary processes in this context. The higher levels recorded for households and catering services highlight the need for greater awareness of culinary processes and for measures to be adopted in these settings to limit the formation of acrylamide in food preparation.

Effect of consumer's decisions on acrylamide exposure during the preparation of French fries. Part 2: Color analysis.

Two observational tests were designed to investigate the influence of consumer decisions during the preparation of French fries from fresh potatoes on acrylamide formation. In both tests, color was of paramount importance to volunteers with regards to the decision to stop the frying process. CIE-Lab* color parameters and visual color assessments of French fries were associated with general acrylamide content and its classification according to the EU benchmark levels (BL, 500 µg/kg). Binary logistic regression analysis revealed that color parameter a* was the main factor in determining the probability of acrylamide content being above BL, regardless of the culinary practices applied. A cut-off of 0.855 for a* produced the greatest sensitivity (81.1%) and specificity (99.9%) for classifying samples with acceptable BL of acrylamide content. An increase of one unit of a* more than quadruple the risk of acrylamide content not falling within BL. When no external color reference was provided, volunteers misclassified 33.6% of fries as having a golden color. Harmonized criteria based on a reference color chart that indicates a golden color are necessary for producing fries with reduced acrylamide contents and, in this way, limit the risk of exposure to acrylamide in domestic settings.

Effect of consumer's decisions on acrylamide exposure during the preparation of French fries. part 1: Frying conditions.

Variability in home-cooking practices affects dietary exposure to acrylamide, limiting risk evaluation. Two observational tests (controlled and randomized) involving 127 volunteers was designed to investigate the influence of consumer decisions on acrylamide formation during the preparation of French fries. Consumer practices were recorded during pre-frying, frying and post-frying stages. Reducing sugar content, asparagine, moisture, acrylamide, moisture, weight loss and color were evaluated. In the controlled assay, mean acrylamide content was 508 µg/kg and volunteers took a mean frying time of 91.9 s (87.5-96.4, C.I. 95%). The narrow confidence interval of 8.9 s represented an increase of 120 µg/kg in the acrylamide content of the fried potato. Average acrylamide content in the randomized assay (215 µg/kg, 150-280 µg/kg, C.I. 95%) was significantly lower than in the controlled assay. Amongst the culinary practices applied for the potato variety evaluated, washing of the potato strips significantly reduced acrylamide content in French fries. The percentage of samples with acrylamide lower than the benchmark value (500 µg/kg) was 54% and 93% for the controlled and randomized assays, respectively. Estimated average dietary exposure to acrylamide from French fries in adults was 5.65 µg/day, which corresponds to a margin of exposure of 2105 for carcinogenicity.

Introduction to the Special Issue: New Frontiers in Acrylamide Study in Foods-Formation, Analysis and Exposure Assessment.

Acrylamide is a chemical contaminant that naturally originates during the thermal processing of many foods. Since 2002, worldwide institutions with competencies in food safety have promoted activities aimed at updating knowledge for a revaluation of the risk assessment of this process contaminant. The European Food Safety Authority (EFSA) ruled in 2015 that the presence of acrylamide in foods increases the risk of developing cancer in any age group of the population. Commission Regulation (EU) 2017/2158 establishes recommended mitigation measures for the food industry and reference levels to reduce the presence of acrylamide in foods and, consequently, its harmful effects on the population. This Special Issue explores recent advances on acrylamide in foods, including a novel insight on its chemistry of formation and elimination, effective mitigation strategies, conventional and innovative monitoring techniques, risk/benefit approaches and exposure assessment, in order to enhance our understanding for this process contaminant and its dietary exposure.

Prebiotic Properties of Non-Fructosylated α-Galactooligosaccharides from PEA (Pisum sativum L.) Using Infant Fecal Slurries.

The interest for naturally-occurring oligosaccharides from plant origin having prebiotic properties is growing, with special focus being paid to supplemented products for infants. Currently, non-fructosylated α-galactooligosaccharides (α-GOS) from peas have peaked interest as a result of their prebiotic activity in adults and their mitigated side-effects on gas production from colonic bacterial fermentation. In this study, commercially available non-fructosylated α-GOS from peas and ß-galactooligosaccharides (ß-GOS) derived from lactose were fermented using fecal slurries from children aged 11 to 24 months old during 6 and 24 h. The modulatory effect of both GOS on different bacterial groups and bifidobacteria species was assessed; non-fructosylated α-GOS consumption was monitored throughout the fermentation process and the amounts of lactic acid and short-chain fatty acids (SCFA) generated were analyzed. Non-fructosylated α-GOS, composed mainly of manninotriose and verbascotetraose and small amounts of melibiose, were fully metabolized and presented remarkable bifidogenic activity, similar to that obtained with ß-GOS. Furthermore, non-fructosylated α-GOS selectively caused an increase on the population of Bifidobacterium longum subsp. longum and Bifidobacterium catenulatum/pseudo-catenulatum. In conclusion, non-fructosylated α-GOS could be used as potential ingredient in infant formula supplemented with prebiotic oligosaccharides.

Are Household Potato Frying Habits Suitable for Preventing Acrylamide Exposure?

A survey was conducted of 730 Spanish households to identify culinary practices which might influence acrylamide formation during the domestic preparation of french fries and their compliance with the acrylamide mitigation strategies described in the 2017/2158 Regulation. Spanish household practices conformed with the majority of recommendations for the selection, storing and handling of potatoes, with the exception of soaking potato strips. Olive oil was the preferred frying oil (78.7%) and frying pans were the most common kitchen utensils used for frying (79.0%), leading to a higher oil replacement rate than with a deep-fryer. Although frying temperature was usually controlled (81.0%), participants were unaware of the maximum temperature recommended for preventing acrylamide formation. For french fries, color was the main criteria when deciding the end-point of frying (85.3%). Although a golden color was preferred by respondents (87.3%), color guidelines are recommended in order to unify the definition of "golden." The results conclude that habits of the Spanish population are in line with recommendations to mitigate acrylamide during french fry preparation. Furthermore, these habits do not include practices that risk increasing acrylamide formation. Nevertheless, educational initiatives tailored towards consumers would reduce the formation of this contaminant and, consequently, exposure to it in a domestic setting.

Association between Heat-Induced Chemical Markers and Ultra-Processed Foods: A Case Study on Breakfast Cereals.

Nutritional composition and neo-formed contaminant content in ultra-processed foods, amongst other factors, may contribute to increasing overall risk of non-communicable diseases and cancer. Commercial breakfast cereals (n = 53) were classified according to the NOVA approach as un-/minimally processed (NOVA-1, 11%), processed (NOVA-3, 30%), and ultra-processed (NOVA-4, 59%) foods. Acrylamide and hydroxymethylfurfural (HMF) content as heat-induced chemical markers was taken from our research team database. The NutriScore was used as the nutritional profiling system. Samples were distributed between groups A (19%), B (13%), C (38%), and D (30%). No statistically significant differences in acrylamide and HMF were found across the NutriScore groups. Sugar content was the only nutritional descriptor found to be significantly different between processed (11.6 g/100 g) and ultra-processed (23.1 g/100 g) breakfast cereal groups. Sugar content correlated with acrylamide (p < 0.001) and HMF (p < 0.0001). Acrylamide and HMF contents were not significantly higher in the NOVA-4 group when compared with the NOVA-3 group. However, trends towards higher acrylamide and HMF content are observed, amounting to a change of 75 µg/kg and 13.3 mg/kg in processed breakfast cereals, and 142 µg/kg and 32.1 mg/kg in ultra-processed breakfast cereals, respectively. Thus, the NOVA classification may not reflect the extent of the thermal treatment applied to the breakfast cereal but the type and amount of ingredients incorporated. Ultra-processed breakfast cereal does not predict significantly higher toxicological concern based on acrylamide content than processed breakfast cereals; a clear trend is seen whose contributing factors should be further studied.

Formation of Acrylamide and other Heat-Induced Compounds during Panela Production.

Non-centrifugal cane sugar (panela) is an unrefined sugar obtained through intense dehydration of sugarcane juice. Browning, antioxidant capacity (measured by ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) assay and total phenolic content) and the formation of acrylamide and other heat-induced compounds such as hydroxymethylfurfural (HMF) and furfural, were evaluated at different stages during the production of block panela. Values ranged between below the limit of quantitation (LOQ)-890 µg/kg, < LOQ-2.37 mg/kg, < LOQ-4.5 mg/kg, 0.51-3.6 Abs 420 nm/g, 0.89-4.18 mg gallic acid equivalents (GAE)/g and 5.08-29.70 µmol TE/g, for acrylamide, HMF, furfural, browning, total phenolic content and ABTS (all data in fresh weight), respectively. Acrylamide significantly increased as soluble solid content increased throughout the process. The critical stages for the formation of acrylamide, HMF and furfural were the concentration of the clarified juice in the concentration stage to get the panela honey and the final stage. Similar trends were observed for the other parameters. This research concludes that acrylamide, HMF and furfural form at a high rate during panela processing at the stage of juice concentration by intense evaporation. Therefore, the juice concentration stage is revealed as the critical step in the process to settle mitigation strategies.

How Far is the Spanish Snack Sector from Meeting the Acrylamide Regulation 2017/2158?

In 2017, the European Commission published Regulation 2017/2158 establishing mitigation measures and benchmark levels to reduce acrylamide in foods. Acrylamide was determined in seventy potato crisp samples commercialized in Spain. The aim was to update knowledge about the global situation in the snack sector and evaluate the effectiveness of mitigation strategies applied, especially since the publication of the Regulation. Results were compared with data previously published in 2004, 2008, and 2014, assessing the evolution over recent years. Average acrylamide content in 2019 (664 µg/kg, range 89-1930 µg/kg) was 55.3% lower compared to 2004, 10.3% lower compared to 2008 and practically similar to results from 2014. Results support the effectiveness of mitigation measures implemented by Spanish potato crisp manufacturers. However, 27% of samples exhibited concentrations above the benchmark level established in the Regulation (750 µg/kg), which suggests that efforts to reduce acrylamide formation in this sector must be continued. Besides the variability seen between samples, acrylamide significantly correlated with the color parameter a*, which enables discrimination of whether potato crisps contain above or below benchmark content. The calculated margin of exposure for carcinogenicity was below the safety limit, which should be considered from a public health point of view.

Acrylamide in French fries prepared at primary school canteens.

Children are one of the most exposed groups to dietary acrylamide with 'potato fried products' that account for up to half the total exposure to this contaminant. Acrylamide was measured in French fries prepared in 31 primary school canteens randomly recruited from different Spanish regions. The mean content was 329 µg kg-1 (from <20 to 4000 µg kg-1). French fries prepared from frozen par-fried potatoes reported a lower acrylamide content than those from fresh potatoes, 229 and 460 µg kg-1, respectively. Only 15.7% of samples were above the benchmark levels established by the EU Regulation 2017/2158 (500 µg kg-1). Significant differences were found according to the color of toasted (2274 µg kg-1), dark-golden (463 µg kg-1), golden (134 µg kg-1) and light-golden (52 µg kg-1) French fries. All the samples that were golden and light-golden showed acrylamide content below the threshold. The chromatic parameter a* was used as a rough classification of the French fries according to the benchmark level. Both educational initiatives intended for food operators and managerial-technical criteria that include the code of frying practices at public food service establishments should consider the golden color as the target for the end-point of frying. Acrylamide exposure will be reduced and, therefore, the risk linked to French fries consumption. This fact is especially relevant in establishments that prepare food for schoolchildren and would help to provide healthier diets, not only from a nutritional point of view but also from the reduction of chemical contaminants.

Skeletal muscle proteome of piglets is affected in a muscle-dependent manner by a limiting total sulfur amino acid supply.

PURPOSE: A total sulfur amino acid (TSAA) deficient diet can affect the amino acid composition of skeletal muscles. However, it is unknown how the different muscle proteins are affected by the TSAA deficiency. METHODS: The proteomic profiles of the fast-twitch glycolytic longissimus (LM) and the slow-twitch oxidative rhomboideus (RM) muscles were compared in 42-day-old piglets fed either a 28% deficient (TSAA-) or a sufficient (TSAA+) diet in TSAA for 10 days. Differentially regulated proteins were identified and submitted to Gene Ontology Pathways Analysis to identify biological processes affected by TSAA deficiency. RESULTS: A total of 36 proteins in LM and 24 proteins in RM differed in abundance between the two dietary treatments. In both muscles, an increased oxidative energy metabolism was observed in TSAA- piglets. However, a greater mitochondrial oxidation of pyruvate generated from glycolysis was observed in LM of TSAA- piglets, whereas fatty acid ß-oxidation and glycogen sparing were favored in RM. This suggests a muscle-specific reorientation of energy metabolism in response to a TSAA- deficiency. In both muscles, the protein abundance and enzyme activity of superoxide dismutase were increased in TSAA- piglets. Other enzymes involved in antioxidant defense, heat shock proteins coping with cellular stress, and annexins involved in the regulation of apoptosis were generally found to be more expressed in the LM of TSAA- piglets, with no or minor changes in RM. CONCLUSIONS: Skeletal muscle proteome in young growing piglets was modulated in a muscle-dependent manner by a deficient TSAA supply, with accentuated changes in fast-twitch glycolytic muscle.

Acrylamide in biscuits commercialised in Spain: a view of the Spanish market from 2007 to 2019.

The biscuit industry has evolved in the last decades with the inclusion of alternative cereals, pseudo-cereals and other ingredients in traditional recipes, which allows current consumers' needs to be met. The dough composition, thermal treatment applied during baking, low moisture, and high rate of consumption make biscuits important contributors to daily exposure to acrylamide in Western diets. Acrylamide was determined by LC-ESI-MS/MS in 80 commercial biscuits marketed in Spain. The mean acrylamide content was 343 µg kg-1 (from <20 to 2144 µg kg-1). Rye, teff and oat-based biscuits exhibited the highest content (2144, 1559 and 1424 µg kg-1, respectively). 70% of the samples were below the benchmark level established by the EU Regulation 2017/2158. Compared to our previous prospective study in 2007 on the Spanish market, acrylamide has experienced an important decline (45%) in wheat-based formulations, demonstrating the effectiveness of the mitigation strategies applied in the sector. However, when all samples were considered, a slight non-significant decrease (18%) was observed. This fact suggests that the reformulation of traditional recipes with innovative cereals and ingredients could increase the acrylamide content in biscuits despite the mitigation strategies applied. Therefore, forthcoming reviews of the regulation should consider establishing categories in biscuits according to the main cereal, as has already been established in breakfast cereals.

Glycation affects differently the main soybean Bowman-Birk isoinhibitors, IBB1 and IBBD2, altering their antiproliferative properties against HT29 colon cancer cells.

Naturally-occurring serine protease inhibitors of the Bowman-Birk family, particularly abundant in legume seeds, exert their potential chemopreventive and/or therapeutic properties via protease inhibition. Processing of legume seeds, including soybeans, has been proposed as a major cause for their loss of bioactivity due to glycation. In order to assess how glycation affected the protease inhibitory activities of major soybean Bowman-Birk isoinhibitors (BBI) and their antiproliferative properties, IBB1 and IBBD2 were purified and subjected to glycation under controlled conditions using glucose at high temperature. Both soybean isoinhibitors showed remarkable heat stability. In the presence of glucose, IBBD2 lost most of its trypsin inhibitory activity while IBB1 maintains similar trypsin and chymotrypsin inhibitory activities as in the absence of sugar. Glycation patterns of both BBI proteins were assessed by MALDI-TOF spectrometry. Our results show that the glycation process affects IBBD2, losing partially its antiproliferative activity against HT29 colon cancer cells, while glycated-IBB1 was unaffected.

Occurrence of acrylamide and other heat-induced compounds in panela: Relationship with physicochemical and antioxidant parameters.

Panela is a natural, unrefined non-centrifugal sugar obtained by intense dehydration of sugarcane juice. Acrylamide, hydroxymethylfurfural (HMF), and furfural were determined in 40 panela samples distributed as granulated and block according to the technological process. Colour, browning, moisture, water activity, pH and antioxidant capacity were also evaluated. Acrylamide ranged between 60 and 3058 µg/kg; granulated panela reporting the highest concentration (812 µg/kg) compared with block panela (540 µg/kg). The lower content in HMF and furfural, the intense dehydration, and the extensive darkening of granulated panela suggested the browning reactions were boosted due to the application of more severe thermal treatments. Principal component analysis showed a significant relationship between the panela presentation and the concentration of the analysed compounds. Benchmark values considering both types of processes would help to establish mitigation initiatives in panela products. The chromatic parameter a* could be used as an indirect index of the acrylamide content in panela.

Occurrence of Furosine and Hydroxymethylfurfural in Breakfast Cereals. Evolution of the Spanish Market from 2006 to 2018.

The demand for healthier products has led the breakfast cereal sector to develop new formulations to improve the nutritional profile of breakfast cereals; however, the increase in chemical risks should also be evaluated. Amadori compounds and 5-hydroxymethylfurfural (HMF) are Maillard reaction products applied as heat damage indices in breakfast cereals. Furosine (a synthetic amino acid formed by acid hydrolysis of Amadori compounds) is linked to the loss of protein quality, while HMF has exhibited toxicological effects in cells and animals. Furosine and HMF content was evaluated in Spanish breakfast cereals whereas the effect of protein, fibre, and sugar content, the type of grain, the presence of honey, and the manufacturing process were discussed, as well as compared with a previous prospective study. The average furosine and HMF contents were 182 mg/kg and 21.7 mg/kg, respectively. Protein and fibre content were directly related to the furosine content, whereas sugar level, honey addition, and the manufacturing process affected the content of HMF. Occurrence of furosine and HMF decreased nearly 40% in a decade (2006-2018). These findings are relevant in terms of nutritional score, since lysine availability is preserved, but also from a toxicological point of view, due to the decreased daily exposure to both compounds, which dropped 30%.