Behavior of diclofenac from contaminated fish after cooking and in vitro digestion.

BACKGROUND: Seafood consumers are widely exposed to diclofenac due to the high contamination levels often present in aquatic organisms. It is a potential risk to public health due its endocrine disruptor properties. Limited information is available about diclofenac behavior after food digestion to enable a more realistic scenario of consumer exposure. This study aimed to evaluate cooking effects on diclofenac levels, and determine diclofenac bioaccessibility by an in vitro digestion assay, using commercial fish species (seabass and white mullet) as models. The production of the main metabolite 4'-hydroxydiclofenac was also investigated. Fish hamburgers were spiked at two levels (150 and 1000 ng g-1) and submitted to three culinary treatments (roasting, steaming and grilling). RESULTS: The loss of water seems to increase the diclofenac levels after cooking, except in seabass with higher levels. The high bioaccessibility of diclofenac (59.1-98.3%) observed in both fish species indicates that consumers' intestines are more susceptible to absorption, which can be worrisome depending on the level of contamination. Contamination levels did not affect the diclofenac bioaccessibility in both species. Seabass, the fattest species, exhibited a higher bioaccessibility of diclofenac compared to white mullet. Overall, cooking decreased diclofenac bioaccessibility by up to 40% in seabass and 25% in white mullet. The main metabolite 4'-hydroxydiclofenac was not detected after cooking or digestion. CONCLUSION: Thus, consumption of cooked fish, preferentially grilled seabass and steamed or baked white mullet are more advisable. This study highlights the importance to consider bioaccessibility and cooking in hazard characterization studies. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Emerging mycotoxins in infant and children foods: A review.

A proper nutrition is crucial for children's healthy development. Regardless of the usual recommendations to follow a varied diet, some foods can be a source of toxic natural contaminants such as mycotoxins, potent secondary metabolites produced by filamentous fungi. In addition to the most well-known mycotoxins, many of which are subject to tight regulation regarding the maximum levels allowed in different types of food, there is a large group of mycotoxins, the so-called emerging mycotoxins, about which less knowledge has already been acquired, which have gradually been the target of interest from the scientific community due to their prevalence in most foodstuffs, particularly in cereals and cereal-based products. Alternariol and his metabolite alternariol mono-methyl ether, beauvericin, citrinin, culmorin, enniatins, ergot alkaloids, fusaproliferin, kojic acid, moniliformin, sterigmatocystin, tentoxin and tenuazonic acid are the most representative of them. The current review gathered the information of the last ten years that have been published on the levels of emerging mycotoxins in food products dedicated for infants and children. European Union countries are responsible for most of the reported studies, which showed levels that can reach hundreds of mg/kg.

Pesticide residues in herbs and their transfer for infusions.

In this work, a cost-effective gas chromatography-mass spectrometry method was validated for the analysis of twenty-five pesticide residues in herbs and infusions using a quick, easy, cheap, effective, rugged, and safe procedure or a dispersive liquid-liquid microextraction method, respectively. Figures of merit of the method showed good accordance with current guidelines. From the 58 herb samples studied (pertaining to 20 different species), 80% presented at least one detectable pesticide, with 62% of them above the maximum residual level. Results showed that pesticide residues from naturally contaminated herbs were not transferred at a significant rate to the herbal infusions. When a control assay was conducted by spiking a blank herb sample with a large amount of each pesticide (7 mg/L) 15 analytes were detected below the limit of quantification in the infusion.

Survey on endocrine-disrupting chemicals in seafood: Occurrence and distribution.

Currently, the presence of endocrine disrupting chemicals (EDCs) in the marine environment pose а potential risk to both wildlife and human health. The occurrence of EDCs in seafood depends of several factors such as source and amounts of EDCs that reach the aquatic environment, physicochemical features of EDCs, and its accumulation in trophic chain. This review highlights the occurrence and distribution of EDCs along the seafood in the last 6 years. The following EDCs were included in this review: brominated flame retardants (PBDEs, PBBs, HBCDDs, TBBPA, and novel flame retardants); pharmaceuticals (paracetamol, ibuprofen, diclofenac, carbamazepine), bisphenols, hormones, personal care products (Musk and UV Filters), and pesticides (organochlorides, organophosphates, and pyrethroids). Some of them were found above the threshold that may cause negative effects on human, animal, and environmental health. More control in some countries, as well as new legislation and inspection over the purchase, sale, use, and production of these compounds, are urgently needed. This review provides data to support risk assessment and raises critical gaps to stimulate and improve future research.

Effects of Green Tea Marinade in the Bioaccessibility of Tonalide and Benzophenone 3 in Cooked European Seabass.

Fish consumption is an indicator of human exposure to personal care products (PCP) such as tonalide (AHTN) and benzophenone 3 (BP3). Although most fish consumed is cooked, the impact of cooking procedures on PCP levels is difficult to evaluate. Hence, the aim of this work was to provide thorough information on the stability and bioaccessibility of AHTN and BP3 upon cooking and in vitro digestion. A green tea (Camellia sinensis) marinade, rich in polyphenol, was used as mitigating strategy to reduce these contaminants. Roasting and frying reduced AHTN and BP3 levels in European seabass (Dicentrarchus labrax) spiked samples. Additionally, the green tea marinade promoted a reduction of up to 47% AHTN and 35% BP3. Bioaccessibility of AHTN was higher (up to 45%), and increased with the use of green tea marinades. BP3 showed a bioaccessibility below 19% in all cooked samples. Overall, a decrease in PCP levels was observed after cooking; this decrease was even more pronounced when marination was previously used. However, this decrease is cancelled out by the fact that the bioaccessible fraction of the contaminants increases in an inverse way; therefore, none of these processes can be considered a mitigating alternative.

Phthalic acid esters and adipates in herbal-based soft drinks: an eco-friendly method.

Phthalic acid esters (PAEs) and adipates are plasticizers with high applicability in several products and building materials (e.g. cosmetics, packing) very persistent in the environment, features which render them ubiquitous pollutants. These substances can contaminate food through the environment (water, air, and soil) and/or migration from packaging materials, which creates a health concern due to their toxicity. This paper describes an eco-friendly dispersive liquid-liquid microextraction (DLLME) procedure to extract five phthalates and bis(2-ethylhexyl) adipate (DEHA) from bottled herbal-based beverages followed by GC-MS/MS quantification. The method showed low limits of detection (5.0-13 µg L-1) and quantification (20-35 µg L-1), good inter- and intraday precision (RSD < 19%), and recoveries ranging from 82 to 111%. It was applied to 16 real samples, of which 13 showed the presence of at least one of the analytes under study. Additionally, an exposure assessment was performed, and resulted in a hazard quotient less than 1 (HQ < 1) for all analytes. Therefore, PAEs and DEHA found in samples do not pose a health issue.

Urinary bisphenol levels in plastic industry workers.

Bisphenol A (BPA) is a known endocrine disruptor compound that is widely applied as a monomer base in polycarbonate plastics and as a binding agent in several epoxy resins. Plastic industry workers have usually heavier and prolonged exposures to BPA. Hence, the present work aims to assess the levels of BPA and their analogs (S, F, B, AF, Z, E, and AP) in 170 urine samples from a cross-sectional study of workers from a plastic industry located in north Constantine (Algeria). This work was complemented with a questionnaire about sexual functions and evaluation of sexual hormone levels. The results showed a stable presence of BPA (average of 3.24 µg/L), accounting for more than 90% of the total BPs. Of the remaining BP analogs, only trace amounts of BPB were detected in three samples (average of 2.73 µg/L). Significant associations with BPA urinary levels were noted with age (p = 0.006), occupational level of exposure (p = 0.023), and years of experience (p = 0.001).

A novel GC-MS methodology to evaluate aromatase activity in human placental microsomes: a comparative study with the standard radiometric assay.

Estrogens are key factors in the development of the estrogen receptor-positive (ER+) breast cancer. Estrogens, estrone (E1), and estradiol (E2) production is achieved by aromatase, a cytochrome P450 enzyme that has androgens, androstenedione (AD), and testosterone (T) as substrates. Nowadays, third-generation aromatase inhibitors (AIs) are considered the gold-standard treatment for ER+ breast cancer in postmenopausal women as well as in premenopausal women with ovary ablation. Aromatase activity assessment still relies on radiometric assays that are expensive, hazardous, and non-environmentally friendly. Thus, in order to overcome these disadvantages, a new methodology was developed to evaluate aromatase activity, based on dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-mass spectrometry (GC-MS). The enzymatic reaction was carried out in human placental microsomes, using AD as substrate, and the anti-aromatase activity was measured by determining the conversion percentage of AD into E1 (ratio E1/AD) using isotopic analogues as internal standards. The method showed good linearity (r2 = 0.9908 for AD and 0.9944 for E1), high accuracy (more than 74% for AD and more than 66% for E1), high extraction efficiency, and good intra-day and inter-day precision (below 14%, 4 levels). In this work, the IC50 values of the third-generation AIs, anastrozole, letrozole, and exemestane, obtained from the radiometric assay are also compared, and similar IC50 values are described. This method is a good alternative to the current radiometric assay, being fast and sensitive with a good extraction efficiency, accuracy, and recovery. In addition, it may be applied for the evaluation of the anti-aromatase activity of new potential AIs. Graphical abstract.

Quantification of eight bisphenol analogues in blood and urine samples of workers in a hazardous waste incinerator.

Bisphenol A (BPA) has been widely used in the manufacture of polycarbonate plastic and epoxy resins. In recent years, producers have started replacing BPA by other chemical analogues, such as bisphenol -S (BPS) and -F (BPF), all of them under the label "BPA-free". However, despite bisphenol (BP) analogues have a very similar structure, their endocrine-disrupting properties could differ from those of BPA. Unfortunately, information regarding human exposure to BP analogues is very limited, not only as single substances, but also as chemical mixtures. The aim of this study was to determine the levels of 8 BP analogues (A, S, F, B, AF, Z, E, and AP) in biological samples from a controlled cohort of workers in a hazardous waste incinerator (HWI) located in Constantí (Catalonia, Spain). Firstly, a chemical method to analyze a mixture of those 8 analogues in total blood and urine was optimized, being samples quantified by means of gas chromatography coupled to mass spectrometry (GC-MS). Furthermore, a biomonitoring study was performed by collecting samples of total blood and urine of 29 people working in the HWI. Among the 8 BP analogues assessed, BPA presented the highest levels in both biological samples, with mean total (free + conjugated) BPA concentrations of 0.58 and 0.86 µg/L in blood and urine, respectively. Free vs. total BPA levels presented a mean percentage of 79% in blood and 19% in urine. Beyond BPA, traces of BPB were also found in a single sample of blood. Furthermore, none of the remaining BP analogues was detected in blood or urine. Despite BPA has been regulated, it is still very present in the environment, being human exposure to this chemical still an issue of concern for the public health.

Impact of potatoes deep-frying on common monounsaturated-rich vegetable oils: a comparative study.

Aiming to distinguish the nutritional and safety impacts on consumer's health of prolonged frying with vegetable oils rich in monounsaturated fatty acids (MUFA), namely peanut oil (PO), canola oil (CO) and extra virgin olive oil (EVOO), a domestic deep-frying assay using fresh potatoes was implemented (175 °C, 8 h per day, up to 28 h). Based on a total polar compounds (TPC) degradation limit of 25%, PO and CO enabled 18-20 h of frying, while EVOO allowed significantly higher frying hours (> 28 h). Despite the non-significant variations in oxidized triglycerides contents observed through time, and loss of all major antioxidants during the first 8 to 12 h of frying, PO showed statistically higher amounts of conjugated dienes (27 at 20 h; against 19 in CO and 17 in EVOO) and CO of anisidine value (252 at 20 h; against 209 in PO and 100 in EVOO), indicative of different oxidation patters. This was corroborated with the analysis of major volatiles, with PO and CO being statistically richer in alkenals and alkadienals, respectively. Therefore, despite the MUFA predominance, differences in their unsaturation profile impact on the type and amount of degradations products formed under prolonged frying and consequently on consumer's health. As to EVOO use for prolonged frying, despite its increased resistance to oxidation and lower risk of formation of unhealthy volatiles, it loses its pool of natural bioactive compounds in the first hours of frying.

Assessing the effects of seawater temperature and pH on the bioaccumulation of emerging chemical contaminants in marine bivalves.

Emerging chemical contaminants [e.g. toxic metals speciation, flame retardants (FRs) and perfluorinated compounds (PFCs), among others], that have not been historically recognized as pollutants nor their toxicological hazards, are increasingly more present in the marine environment. Furthermore, the effects of environmental conditions (e.g. temperature and pH) on bioaccumulation and elimination mechanisms of these emerging contaminants in marine biota have been poorly studied until now. In this context, the aim of this study was to assess, for the first time, the effect of warmer seawater temperatures (Δ = + 4°C) and lower pH levels (Δ = - 0.4 pH units), acting alone or combined, on the bioaccumulation and elimination of emerging FRs (dechloranes 602, 603 and 604, and TBBPA), inorganic arsenic (iAs), and PFCs (PFOA and PFOS) in two estuarine bivalve species (Mytilus galloprovincialis and Ruditapes philippinarum). Overall, results showed that warming alone or combined with acidification promoted the bioaccumulation of some compounds (i.e. dechloranes 602, 604, TBBPA), but also facilitated the elimination of others (i.e. iAs, TBBPA). Similarly, lower pH also resulted in higher levels of dechloranes, as well as enhanced iAs, PFOA and PFOS elimination. Data also suggests that, when both abiotic stressors are combined, bivalves' capacity to accumulate contaminants may be time-dependent, considering significantly drastic increase observed with Dec 602 and TBBPA, during the last 10 days of exposure, when compared to reference conditions. Such changes in contaminants' bioaccumulation/elimination patterns also suggest a potential increase of human health risks of some compounds, if the climate continues changing as forecasted. Therefore, this first study pointed out the urgent need for further research on the effects of abiotic conditions on emerging contaminants kinetics, to adequately estimate the potential toxicological hazards associated to these compounds and develop recommendations/regulations for their presence in seafood, considering the prevailing environmental conditions expected in tomorrow's ocean.

Domestic Cooking of Muscle Foods: Impact on Composition of Nutrients and Contaminants.

Meat and fish are muscle foods rich in valuable nutrients, such as high-quality proteins, vitamins, and minerals, and, in the case of fish, also unsaturated fatty acids. The escalation of meat and fish production has increased the occurrence of pesticide and antibiotic residues, as result of pest control on feed crops, and antibiotics used to fight infections in animals. Meat and fish are usually cooked to enrich taste, soften texture, increase safety, and improve nutrient digestibility. However, the impact of cooking on nutritional properties and formation of deleterious compounds must be understood. This review summarizes studies, published in the last decade, that have focused on how domestic cooking affects: (i) composition of nutrients (protein, fatty acids, vitamins, and minerals); (ii) antibiotic and pesticide residue contents; and (iii) the formation of cooking-induced contaminants (heterocyclic aromatic amines, polycyclic aromatic hydrocarbons, and thermal degradation products of antibiotics and pesticides). Cooking affects the nutritional composition of meat and fish; frying is the cooking method that causes the greatest impact. Cooking may reduce the pesticide and antibiotic residues present in contaminated raw meat and fish; however, it may result in the formation of degradation products of unknown identity and toxicity. Control of cooking time and temperature, use of antioxidant-rich marinades, and avoiding the dripping of fat during charcoal grilling can reduce the formation of cooking-induced contaminants.

Development of QuEChERS-based extraction and liquid chromatography-tandem mass spectrometry method for simultaneous quantification of bisphenol A and tetrabromobisphenol A in seafood: fish, bivalves, and seaweeds.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of bisphenol A (BPA) and tetrabromobisphenol A (TBBPA) in different seafood samples was developed and validated. Sample preparation was based on a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based procedure through an extraction of target analytes with acidified acetonitrile (MeCN) added with inorganic salts (MgSO4, NaCl) followed by a liquid-liquid extraction (LLE) using hexane-tertbutylmethyl ether/hexane-benzene to eliminate matrix co-extracts. The developed method promotes a better removal of interferences than that achieved with the classic QuEChERS procedure. The method was validated following the guidelines of the European Union (EU) for relevant seafood matrices such as fish, mussel, and seaweed. Accuracy (81 % average of recovery), reproducibility (12 % average relative standard deviation for both intra-day and inter-day repeatability), and sensitivity for the target analytes (method detection limits of 0.07 ng/g wet weight (ww) and 0.06 ng/g ww for BPA and TBBPA, respectively) were evaluated for all the matrices studied. Graphical Abstract Simultaneous analysis of TBBPA and BPA in seaweeds, mussels and fish.

Advances in isoflavone profile characterisation using matrix solid-phase dispersion coupled to HPLC/DAD in Medicago species.

INTRODUCTION: Analytical methods used in phytochemistry analysis are limited by the sample preparation step, which should ideally be fast, accurate, ecofriendly and achievable using low quantities of the sample. Matrix solid-phase dispersion (MSPD) may be a good alternative for combining extraction and purification procedures, thereby reducing the indicated limitations. OBJECTIVE: Applying an MSPD extraction procedure coupled to high-performance liquid chromatography diode-array detection (HPLC/DAD) as an alternative methodology to evaluate isoflavone profiles. METHODS: Isoflavone profiles were determined for the leaves of nine species of Medicago in the late flower phenological stage (one or more nodes with 50% open flowers, no seed pods). Extraction was performed following MSPD, and isoflavone profiles were characterised using HPLC/DAD. The quantified amounts were compared with previous results in different species commonly recognised as good sources of isoflavones. RESULTS: Formononetin was the major isoflavone in most species, except M. polymorpha and M. truncatula. The isoflavone amounts were significantly different among the assayed species, with M. orbicularis and M. arabica as the major isoflavone sources, while M. rigidula presented the lowest contents. Furthermore, the detected differences allow electing the best species as a primary source of a specific isoflavone. CONCLUSION: The MSPD allowed good extraction efficiency, reproducibility and recovery. Some of the species showed relevant isoflavone contents, even when compared with acknowledged plant sources such as soy or red clover. To the best of our knowledge the results presented are reported for the first time in these species.

Investigating the individual and mixture cytotoxicity of co-occurring aflatoxin B1, enniatin B, and sterigmatocystin on gastric, intestinal, hepatic, and renal cellular models.

This study investigates the individual and combined effects of the mycotoxins, Aflatoxin B1 (AFB1), Enniatin B (ENNB) and Sterigmatocystin (STG), on the cellular viability of gastric (NCI-N87), intestinal (Caco-2), hepatic (Hep-G2) and renal (Hek-293) cells, shedding light on synergistic or antagonistic effects using a constant ratio combination design proposed by Chou-Talalay. These toxins are prevalent in cereal-based foods, frequently consumed by children which raises concerns about their exposure to these mycotoxins. This population is particularly vulnerable to the effects of these toxins due to their underdeveloped organs and incompletely structured physiological processes. Results showed that ENB was the most toxic of the three mycotoxins across all cell lines, while STG and AFB1 showed lower toxicity. The combination of ENNB + STG was found to be the most potent in terms of binary mixtures. In regard to ternary combinations, Caco-2 cells are more sensitive to the tested mycotoxins, whereas NCI-N87 cells show lower levels of cell damage. Worrying dose reduction values (>10-fold) were found for ENNB in binary and ternary combinations at low exposure levels. These findings are significant for establishing initial reference values, which play a pivotal role in estimating reference doses that are subsequently incorporated into the broader risk assessment process.

In Vitro Digestion and Intestinal Absorption of Mycotoxins Due to Exposure from Breakfast Cereals: Implications for Children's Health.

Breakfast cereals play a crucial role in children's diets, providing essential nutrients that are vital for their growth and development. Children are known to be more susceptible than adults to the harmful effects of food contaminants, with mycotoxins being a common concern in cereals. This study specifically investigated aflatoxin B1 (AFB1), enniatin B (ENNB), and sterigmatocystin (STG), three well-characterized mycotoxins found in cereals. The research aimed to address existing knowledge gaps by comprehensively evaluating the bioaccessibility and intestinal absorption of these three mycotoxins, both individually and in combination, when consumed with breakfast cereals and milk. The in vitro gastrointestinal method revealed patterns in the bioaccessibility of AFB1, ENNB, and STG. Overall, bioaccessibility increased as the food progressed from the stomach to the intestinal compartment, with the exception of ENNB, whose behavior differed depending on the type of milk. The ranking of overall bioaccessibility in different matrices was as follows: digested cereal > cereal with semi-skimmed milk > cereal with lactose-free milk > cereal with soy beverage. Bioaccessibility percentages varied considerably, ranging from 3.1% to 86.2% for AFB1, 1.5% to 59.3% for STG, and 0.6% to 98.2% for ENNB. Overall, the inclusion of milk in the ingested mixture had a greater impact on bioaccessibility compared to consuming the mycotoxins as a single compound or in combination. During intestinal transport, ENNB and STG exhibited the highest absorption rates when ingested together. This study highlights the importance of investigating the combined ingestion and transport of these mycotoxins to comprehensively assess their absorption and potential toxicity in humans, considering their frequent co-occurrence and the possibility of simultaneous exposure.

Volatilome, Microbial, and Sensory Profiles of Coffee Leaf and Coffee Leaf-Toasted Maté Kombuchas.

Kombucha is a fermented beverage traditionally made from the leaves of Camelia sinensis. The market has drastically expanded recently, and the beverage has become more elaborated with new, healthy food materials and flavors. Pruning and harvesting during coffee production may generate tons of coffee leaves that are discarded although they contain substantial amounts of bioactive compounds, including those found in maté tea and coffee seeds. This study characterized the changes in volatilome, microbial, and sensory profiles of pure and blended arabica coffee leaf tea kombuchas between 3-9 days of fermentation. Acceptance was also evaluated by consumers from Rio de Janeiro (n = 103). Kombuchas (K) were prepared using black tea kombucha starter (BTKS) (10%), sucrose (10%), a symbiotic culture of Bacteria and Yeasts (SCOBY) (2.5%), and a pure coffee leaf infusion (CL) or a 50:50 blend with toasted maté infusion (CL-TM) at 2.5%. The RATA test was chosen for sensory profile characterization. One hundred volatile organic compounds were identified when all infusions and kombucha samples were considered. The potential impact compounds identified in CL K and CL-TM K were: methyl salicylate, benzaldehyde, hexanal, nonanal, pentadecanal, phenylethyl-alcohol, cedrol, 3,5-octadien-2-one, ß-damascenone, α-ionone, ß-ionone, acetic acid, caproic acid, octanoic acid, nonanoic acid, decanoic acid, isovaleric acid, linalool, (S)-dihydroactinidiolide, isoamyl alcohol, ethyl hexanoate, and geranyl acetone. Aroma and flavor descriptors with higher intensities in CL K included fruity, peach, sweet, and herbal, while CL-TM K included additional toasted mate notes. The highest mean acceptance score was given to CL-TM K and CL K on day 3 (6.6 and 6.4, respectively, on a nine-point scale). Arabica coffee leaf can be a co-product with similar fingerprinting to maté and black tea, which can be explored for the elaboration of potentially healthy fermented beverages in food industries.

Dynamics and effects of plastic contaminants' assimilation in gulls.

Polybrominated diphenyl ethers are persistent disrupters assimilated by organisms, yet little is known about their link to plastic ingestion and health effects. In an experiment, two groups of yellow-legged/lesser black-backed gulls (Larus michahellis/Larus fuscus) were fed plastics with BDE99 to assess leaching into brain, preen oil, liver and fat tissues and evaluate effects on health and stress parameters. Although most plastic was regurgitated, we observed a clear relation between plastic ingestion and chemical leaching. BDE99 exhibited higher levels in brain tissue of gulls from the plastic groups. Also, only values of cholinesterases measured in plasma were significantly reduced in the 'plastic' groups. Cholinesterase activity in the brain also tended to decrease, suggesting a negative effect in gulls' neurofunction. Results indicate that chemical leaching occurs, even when plastics stay in the stomach for a short period of time and showed that this can affect gulls' health.

Evaluating the human neurotoxicity and toxicological interactions impact of co-occurring regulated and emerging mycotoxins.

Mycotoxins can inflict harmful effects on diverse organs, and mounting evidence indicates their potential involvement in human neurodegenerative diseases. Given the common occurrence of these toxins in food, there is an increasing demand for a comprehensive assessment of their combined toxicity to enhance our understanding of their potential hazards. This research investigates mycotoxin exposure from widely consumed cereal-based products, including enniatin B (ENNB), sterigmatocystin (STG), aflatoxin B1 (AFB1), cyclopiazonic acid (CPZ), citrinin (CIT), and ochratoxin A (OTA). Employing the median-effect equation based on Chou and Talalay's mass-action law, we assessed their cytotoxicity in human SH-SY5Y neuronal cells. Notably, ENNB displayed the highest neurotoxicity (IC50 = 3.72 µM), followed by OTA (9.10 µM) and STG (9.99 µM). The combination of OTA + STG exhibited the highest toxicity (IC50 = 3.77 µM), while CPZ + CIT showed the least detrimental effect. Approximately 70 % of tested binary combinations displayed synergistic or additive effects, except for ENNB + STG, ENNB + AFB1, and CPZ + CIT, which showed antagonistic interactions. Intriguingly, the senary combination displayed moderate antagonism at the lowest exposure and moderate synergism at higher doses. OTA exhibited predominantly synergistic interactions, comprising approximately 90 %, a noteworthy finding considering its prevalence in food. Conversely, ENNB interactions tended to be antagonistic. The most remarkable synergy occurred in the STG and CIT combination, enabling a 50-fold reduction in CIT dosage for an equivalent toxic effect. These findings highlight the biological relevance of robust synergistic interactions, emphasizing the need to assess human exposure hazards accurately, particularly considering frequent mycotoxin co-occurrence in environmental and food settings.

Chemical composition and sensory profiling of coffees treated with asparaginase to decrease acrylamide formation during roasting.

Acrylamide is an amide formed in the Maillard reaction, with asparagine as the primary amino acid precursor. The intake of large amounts of acrylamide has induced genotoxic and carcinogenic effects in hormone-sensitive tissues of animals. The enzime asparaginase is one of the most effective methods for lowering the formation of acrylamide in foods such as potatoes. However, the reported sensory outcomes for coffee have been unsatisfactory so far. This study aimed to produce coffees with reduced levels of acrylamide by treating them with asparaginase while retaining their original sensory and bioactive profiles. Three raw samples of Coffea arabica, including two specialty coffees, and one of Coffea canephora were treated with 1000, 2000, and 3000 ASNU of the enzyme. Asparagine and bioactive compounds (chlorogenic acids-CGA, caffeine, and trigonelline) were quantified in raw and roasted beans by HPLC and LC-MS, while the determination of acrylamide and volatile organic compounds was performed in roasted beans by CG-MS. Soluble solids, titratable acidity, and pH were also determined. Professional cupping by Q-graders and consumer sensory tests were also conducted. Results were analyzed by ANOVA-Fisher, MFA, PCA and Cluster analyses, with significance levels set at p ≤ 0.05. Steam treatment alone decreased acrylamide content by 18.4%, on average, and 6.1% in medium roasted arabica and canefora coffees. Average reductions of 32.5-56.0% in acrylamide formation were observed in medium roasted arabica beans when 1000-3000 ASNU were applied. In the canefora sample, 59.4-60.7% reductions were observed. However, steam treatment primarily caused 17.1-26.7% reduction of total CGA and lactones in medium roasted arabica samples and 13.9-22.0% in canefora sample, while changes in trigonelline, caffeine, and other evaluated chemical parameters, including the volatile profiles were minimal. Increasing enzyme loads slightly elevated acidity. The only sensory changes observed by Q-graders and or consumers in treated samples were a modest increase in acidity when 3000 ASNU was used in the sample with lower acidity, loss of mild off-notes in control samples, and increased perception of sensory descriptors. The former was selected given the similarity in chemical outcomes among beans treated with 2000 and 3000 ASNU loads.