Nutritional profiling, fiber content and in vitro bioactivities of wheat-based biscuits formulated with novel ingredients.

This study evaluated the nutritional profile and fiber content of innovative formulations of wheat-based biscuits enriched with chia seeds, carob flour and coconut sugar. The in vitro antioxidant, cytotoxic, anti-inflammatory and antimicrobial activities were also investigated to understand the potential health advantages of the incorporation of these new ingredients. The novel biscuits demonstrated significant improvements in protein and mineral content, with increases of 50% and 100% in chia biscuits, and up to 20% and 40% in carob biscuits, respectively. Fiber also notably increased, particularly in samples containing 10% carob flour, which increased four times as compared to wheat-based samples. The new ingredients exhibited antibacterial and antifungal activity, particularly against Yersinia enterocolitica (minimum inhibitory concentration 1.25 mg mL-1 in coconut sugar) and Aspergillus fumigatus (minimum inhibitory concentration/minimum fungicidal concentrations 2.5/5 mg mL-1 in chia seeds). However, the final biscuits only displayed antifungal properties. Carob flour and chia seeds had a remarkably high capacity to inhibit the formation of TBARS and promoted greater antioxidant activity in biscuit formulations, with EC50 values decreasing from 23.25 mg mL-1 (control) to 4.54 mg mL-1 (15% defatted ground chia seeds) and 1.19 mg mL-1 (10% carob flour). Only chia seeds exhibited cellular antioxidant, anti-inflammatory and cytotoxic activity, attributes that were lost when seeds were added into the biscuits. These findings highlight the potential health benefits of these ingredients, particularly when incorporated in new wheat-based formulations.

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.

Risk/Benefit Evaluation of Chia Seeds as a New Ingredient in Cereal-Based Foods.

Chia seed (Salvia hispanica L.) is a food rich in protein, fiber, polyunsaturated fatty acids and antioxidants. Consequently, its incorporation in food formulations may be desirable from a nutritional and healthy point of view. However, there is concern regarding the formation of process contaminants when they are subjected to thermal processing. The objective of this study was to incorporate different amounts of ground chia seeds in a biscuit model to evaluate the effect on the antioxidant capacity and formation of acrylamide and furfurals. Seven standard "Maria-type" biscuit formulations were prepared, replacing wheat flour with different amounts of ground chia seeds (defatted and non-defatted), from 0% (control biscuit) to 15% (respect to total solids in the recipe). Samples were baked at 180 °C for 22 min. Compared with the control biscuit, chia formulations increased the content of nutrients, antioxidant capacity (ABTS) and phenolic compounds (Folin-Ciocalteau method) but also doubled acrylamide levels and even raised more than 10 times furanic compound concentrations. Results indicate that the use of chia seeds as ingredients in new cereal-based formulations would improve the nutritional profile but also increase the occurrence of chemical process contaminants. This paradox should be carefully considered in the context of risk/benefit analysis.

Formation of process contaminants in commercial and homemade deep-fried breadcrumbs.

Breading is a culinary technique widespread throughout the world. Deep-frying breaded foods forms a palatable crust but also promotes the formation of compounds of toxicological relevance. The influence of the composition of breadcrumbs on the risk associated with the formation of acrylamide, hydroxymethylfurfural (HMF), and furfural was investigated in a deep-fried breadcrumb coating model. Commercial (CBC) and homemade (HBC) wheat-based breadcrumbs were characterized by the reducing sugars and the asparagine content among other parameters (moisture, pH, and CIELab color). The formation of process contaminants in fried breadcrumbs were not influenced by their initial content, but they were affected by the precursors level. The HMF content was significantly higher (1.4 times) in fried HBC (172 mg/kg) than in CBC (120 mg/kg). By contrast, the acrylamide content was 3 times higher in fried CBC (332 µg/kg) than in HBC (111 µg/kg). Multivariate analysis shows that asparagine is the limiting factor for acrylamide formation, and the reducing sugar content is the main determinant for the formation of furanic compounds. A signal value of 463 µg/kg is proposed for the acrylamide content in the coating of deep-fried breaded foods. The reducing sugars and asparagine content in breadcrumb coatings should be considered when designing breaded foods, thereby reducing the formation and consequently the dietary exposure to these potentially harmful compounds.

Formation of acrylamide during the roasting of chia seeds (Salvia hispanica L.).

Chia is a novel food rich in health-promoting constituents. There is concern regarding the formation of process contaminants when subjected to thermal processing. Acrylamide formation in chia seeds under different roasting conditions (160-200 °C/5-30 min) and formats (whole seeds and ground seeds) was investigated. Acrylamide increased with the intensity of the thermal treatment until a maximum at 180 °C/15 min where levels of precursors were significantly reduced. The fate of acrylamide formation also depended on the physical integrity of the chia seed. Acrylamide formed rapidly on the coat of the seed but represents only 20-25 % of the total. Acrylamide was generated progressively in the inner part of the cell and efficiently retained by the pericarp. The particle size and integrity of chia seeds should be considered in food applications subjected to thermal treatment and grinding or the addition of ground seeds should be revised to reduce the exposure of acrylamide to consumers.

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.

Evaluation of the Olfactory Quality of Roasted Coffee Beans Using a Digital Nose.

The roasting process is one of the critical points to obtain a product of the highest quality with certain sensorial properties including the aroma of coffee. Samples of coffee beans were roasted at different thermal treatment intensities with the aim of obtaining aromatic compounds detected with an electronic device. Sensory analysis, volatile compound profiling, and electronic nose analysis were carried out. Through principal component analysis (95.8% of the total variance of the data was explained by PC1 and PC2) and partial least squares discriminant analysis (the sum of the diagonal elements gave a hit rate of 94%), it could be demonstrated that the E-nose is able to discriminate roasted coffee beans subjected to different thermal treatments. Aromatic profiling was carried out by a testing panel and volatile compounds (VOCs) for the discrimination of roasted coffee samples. Alcohols, aromatics, esters, ketones and furanone were found in higher proportions in samples at the lowest thermal treatment. The VOCs with positive attributes were 1-(4-nitrophenyl)-3-phenylamino-propenone, carboxylic acids, 2-methoxy-4-vinylphenol, and 2-phenylethyl alcohol, while the compounds with negative ones were 2-methyl-furan, 2,5-dimethyl-pyridine, 2-methyl-butanal, and 2-furfurylthiol. The PLS model allows for the quantification of the positive and negative aromas (RCV2 = 0.92) of roasted coffee by using the E-nose. Therefore, the E-nose, that is, an inexpensive and nondestructive instrument, could be a chemometric tool able to discriminate between different qualities of coffee during processing.

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.

Industrial Strategies to Reduce Acrylamide Formation in Californian-Style Green Ripe Olives.

Acrylamide, a compound identified as a probable carcinogen, is generated during the sterilization phase employed during the processing of Californian-style green ripe olives. It is possible to reduce the content of this toxic compound by applying different strategies during the processing of green ripe olives. The influence of different processing conditions on acrylamide content was studied in three olives varieties ("Manzanilla de Sevilla", "Hojiblanca", and "Manzanilla Cacereña"). Olives harvested during the yellow-green stage presented higher acrylamide concentrations than green olives. A significant reduction in acrylamide content was observed when olives were washed with water at 25 °C for 45 min (25% reduction) and for 2 h (45% reduction) prior to lye treatment. Stone olives had 21-26% higher acrylamide levels than pitted olives and 42-50% higher levels than sliced olives in the three studied varieties. When calcium chloride (CaCl2) was added to the brine and brine sodium chloride (NaCl) increased from 2% to 4%, olives presented higher concentrations of this contaminant. The addition of additives did not affect acrylamide levels when olives were canned without brine. Results from this study are very useful for the table olive industry to identify critical points in the production of Californian-style green ripe olives, thus, helping to control acrylamide formation in this foodstuff.

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.

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.

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.