Effect of Acrylamide and Mycotoxins in SH-SY5Y Cells: A Review.

Thermal processes induce the formation of undesired toxic components, such as acrylamide (AA), which has been shown to induce brain toxicity in humans and classified as Group 2A by the International Agency of Research in Cancer (IARC), as well as some mycotoxins. AA and mycotoxins' toxicity is studied in several in vitro models, including the neuroblastoma cell line model SH-SY5Y cells. Both AA and mycotoxins occur together in the same food matrix cereal base (bread, pasta, potatoes, coffee roasting, etc.). Therefore, the goal of this review is to deepen the knowledge about the neurological effects that AA and mycotoxins can induce on the in vitro model SH-SY5Y and its mechanism of action (MoA) focusing on the experimental assays reported in publications of the last 10 years. The analysis of the latest publications shows that most of them are focused on cytotoxicity, apoptosis, and alteration in protein expression, while others are interested in oxidative stress, axonopathy, and the disruption of neurite outgrowth. While both AA and mycotoxins have been studied in SH-SY5Y cells separately, the mixture of them is starting to draw the interest of the scientific community. This highlights a new and interesting field to explore due to the findings reported in several publications that can be compared and the implications in human health that both could cause. In relation to the assays used, the most employed were the MTT, axonopathy, and qPCR assays. The concentration dose range studied was 0.1-10 mM for AA and 2 fM to 200 µM depending on the toxicity and time of exposure for mycotoxins. A healthy and varied diet allows the incorporation of a large family of bioactive compounds that can mitigate the toxic effects associated with contaminants present in food. Although this has been reported in some publications for mycotoxins, there is still a big gap for AA which evidences that more investigations are needed to better explore the risks for human health when exposed to AA and mycotoxins.

Individual and combined effect of acrylamide, fumitremorgin C and penitrem A on human neuroblastoma SH-SY5Y cells.

Acrylamide (AA) is a chemical compound that can be formed in certain foods during high-temperature cooking processes such as frying, baking, and roasting. Exposure to AA has been linked to several neurological effects, including peripheral neuropathy, ataxia, and impaired cognitive function. Penitrem A (PEN A) and Fumitremorgin C (FTC) are toxic mycotoxins produced by certain species of fungi, such as Penicillium Crustosum, Aspergillus Fumigatus and Neosartorya Fischeri. Both mycotoxins are commonly found in contaminated foods and animal feeds and have been linked to several adverse health effects in humans and animals, including the ability to disrupt normal functioning of the nervous system, tremors, seizures, muscle spasms, and convulsions. AA, PEN A, and FTC are all chemical contaminants. Understanding their toxicity and how they may affect human cells can help food safety authorities to establish safe exposure levels for these compounds through food and develop strategies to reduce their presence. The aim of this study was to explore the combined in vitro toxicological effects of AA, PEN A and FTC in SH-SY5Y cells. For this purpose, cells were treated with AA, FTC, and PEN A as an individual and combined treatment. The types of interactions were assessed by the isobologram analysis. The cell cycle was performed by flow cytometry. Additive effect in binary and tertiary combinations was the major effect according to isobologram graphics. Our results demonstrate that PEN A possessed the highest potential in disturbing cell cycle progression by disrupting cell density in G0/G1 phase.

The Occurrence and Health Risk Assessment of Aflatoxin M1 in Raw Cow Milk Collected from Tunisia during a Hot Lactating Season.

Milk is a staple food that is essential for human nutrition because of its high nutrient content and health benefits. However, it is susceptible to being contaminated by Aflatoxin M1 (AFM1), which is a toxic metabolite of Aflatoxin B1 (AFB1) presented in cow feeds. This research investigated AFM1 in Tunisian raw cow milk samples. A total of 122 samples were collected at random from two different regions in 2022 (Beja and Mahdia). AFM1 was extracted from milk using the QuEChERS method, and contamination amounts were determined using liquid chromatography (HPLC) coupled with fluorescence detection (FD). Good recoveries were shown with intra-day and inter-day precisions of 97 and 103%, respectively, and detection and quantification levels of 0.003 and 0.01 µg/L, respectively. AFM1 was found in 97.54% of the samples, with amounts varying from values below the LOQ to 197.37 µg/L. Lower AFM1 was observed in Mahdia (mean: 39.37 µg/L), respectively. In positive samples, all AFM1 concentrations exceeded the EU maximum permitted level (0.050 µg/L) for AFM1 in milk. In Tunisia, a maximum permitted level for AFM1 in milk and milk products has not been established. The risk assessment of AFM1 was also determined. Briefly, the estimated intake amount of AFM1 by Tunisian adults through raw cow milk consumption was 0.032 µg/kg body weight/day. The Margin of Exposure (MOE) values obtained were lower than 10,000. According to the findings, controls as well as the establishment of regulations for AFM1 in milk are required in Tunisia.

Mineral Bioaccessibility and Antioxidant Capacity of Protein Hydrolysates from Salmon (Salmo salar) and Mackerel (Scomber scombrus) Backbones and Heads.

Information on the bioaccessibility of minerals is essential to consider a food ingredient as a potential mineral fortifier. In this study, the mineral bioaccessibility of protein hydrolysates from salmon (Salmo salar) and mackerel (Scomber scombrus) backbones and heads was evaluated. For this purpose, the hydrolysates were submitted to simulated gastrointestinal digestion (INFOGEST method), and the mineral content was analyzed before and after the digestive process. Ca, Mg, P, Fe, Zn, and Se were then determined using an inductively coupled plasma spectrometer mass detector (ICP-MS). The highest bioaccessibility of minerals was found in salmon and mackerel head hydrolysates for Fe (≥100%), followed by Se in salmon backbone hydrolysates (95%). The antioxidant capacity of all protein hydrolysate samples, which was measured by Trolox Equivalent Antioxidant Capacity (TEAC), increased (10-46%) after in vitro digestion. The heavy metals As, Hg, Cd, and Pb were determined (ICP-MS) in the raw hydrolysates to confirm the harmlessness of these products. Except for Cd in mackerel hydrolysates, all toxic elements were below the legislation levels for fish commodities. These results suggest the possibility of using protein hydrolysates from salmon and mackerel backbones and heads for food mineral fortification, as well as the need to verify their safety.

Impact of Combined Processes Involving Ultrasound and Pulsed Electric Fields on ENNs, and OTA Mitigation of an Orange Juice-Milk Based Beverage.

In recent years, several innovative food processing technologies such as ultrasound (USN) and pulsed electric fields (PEF) have emerged in the market, showing a great potential both alone and in combination for the preservation of fresh and processed products. Recently, these technologies have also shown promising applications to reduce mycotoxin levels in food products. Therefore, the objective of this study is to investigate the potential of the combined treatments USN + PEF and PEF + USN on the reduction of Ochratoxin A (OTA) and Enniatins (ENNs) of an orange juice mixed with milk beverage. For this purpose, the beverages were elaborated in the laboratory and individually spiked with mycotoxins at a concentration of 100 µg/L. They were then treated by PEF (30 kV, 500 kJ/Kg) and USN (20 kHz, 100 W, at a maximum power for 30 min). Finally, mycotoxins were extracted using dispersive liquid-liquid microextraction (DLLME), and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS-IT) was employed to determine them. The results showed promising applications, with reductions up to 50% for OTA and up to 47% for Enniatin B (ENNB) after the PEF + USN treatment combination. Lower reduction rates, up to 37%, were obtained with the USN + PEF combination. In conclusion, the combination of USN and PEF technologies could be a useful tool to reduce mycotoxins in fruit juices mixed with milk.

Pulsed electric field (PEF) recovery of biomolecules from Chlorella: Extract efficiency, nutrient relative value, and algae morphology analysis.

This study investigated the effects of pulsed electric field (PEF) (3 kV/cm, 44 pulses, 99 kJ/kg), solvent (H2O or 50 % DMSO) and time (0, 10, 20, 30, 60, 90, 120 and 180 min) on the extraction of Chlorella antioxidant biomolecules and minerals. The results showed that PEF treatment increased the biomolecules recovery. For the extraction time of 120 min, more proteins and polyphenols were obtained using water, while more chlorophyll a and b, and carotenoids were obtained using 50 % DMSO as the extraction solvent. The extracts mineral concentration (PEF vs control) were analysed including Mg, P, Ca, Fe and Zn, and the Relative Nutrient Values results indicated that Chlorella H2O-extracts could be used as a mineral source for different populations. Finally, the fluorescence and scanning electron microscopy revealed the electroporation effect of PEF.

Mycotoxins occurrence in medicinal herbs dietary supplements and exposure assessment.

The multimycotoxin analysis of aflatoxins (AFs), zearalenone (ZEA), ochratoxin A (OTA), enniatins (ENNs) and beauvericin (BEA) was performed in 85 samples of medicinal herbs dietary supplements. The samples were classified in 64 samples of one herbal ingredient and 21 mixed samples. The extraction was performed by QuEChERS method and the determination by liquid chromatography coupled to ion-trap tandem mass spectrometry (LC-MS/MS-IT). Then, the risk characterization to mycotoxins through the consumption of medicinal herbs dietary supplements was assessed. The results showed that ZEA, OTA, ENNs and BEA showed in the samples with incidences between 1 and 34%, being ENNB the most detected mycotoxin. Mycotoxins contents ranged from LOQ to 3850.5 µg/kg while the mean of positives samples were 65.5 µg/kg (ENNA), 82.7 µg/kg (ENNA1), 88.7 µg/kg (ENNB), 324.9 µg/kg (ENNB1), 137.9 µg/kg (BEA) and 1340.11 µg/kg (ZEA), respectively. OTA was detected in one herbal mix tablet for insomnia at concentration of 799 µg/kg. In herbal drugs the European Pharmacopoeia Commission has implemented limits of 2 µg/kg for AFB1 and 4 µg/kg for total AFs. In the present study AFs have not been detected in the analyzed medicinal herbs dietary supplements. The Estimated Daily Intakes (EDIs) values were calculated using a deterministic method, considering two exposure scenarios (lower bound (LB) and upper bound (UB)). The values obtained were in general far below the Tolerable Daily Intakes (TDIs) established. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05306-y.

In Vitro and In Vivo Regulation of SRD5A mRNA Expression of Supercritical Carbon Dioxide Extract from Asparagus racemosus Willd. Root as Anti-Sebum and Pore-Minimizing Active Ingredients.

Oily skin from overactive sebaceous glands affects self-confidence and personality. There is report of an association between steroid 5-alpha reductase gene (SRD5A) expression and facial sebum production. There is no study of the effect of Asparagus racemosus Willd. root extract on the regulation of SRD5A mRNA expression and anti-sebum efficacy. This study extracted A. racemosus using the supercritical carbon dioxide fluid technique with ethanol and investigated its biological compounds and activities. The A. racemosus root extract had a high content of polyphenolic compounds, including quercetin, naringenin, and p-coumaric acid, and DPPH scavenging activity comparable to that of the standard L-ascorbic acid. A. racemosus root extract showed not only a significant reduction in SRD5A1 and SRD5A2 mRNA expression by about 45.45% and 90.86%, respectively, but also a reduction in the in vivo anti-sebum efficacy in male volunteers, with significantly superior percentage changes in facial sebum production and a reduction in the percentages of pore area after 15 and 30 days of treatment. It can be concluded that A. racemosus root extract with a high content of polyphenol compounds, great antioxidant effects, promising downregulation of SRD5A1 and SRD5A2, and predominant facial sebum reduction and pore-minimizing efficacy could be a candidate for an anti-sebum and pore-minimizing active ingredient to serve in functional cosmetic applications.

Nutritional and bioactive oils from salmon (Salmo salar) side streams obtained by Soxhlet and optimized microwave-assisted extraction.

The efficiency of the microwave-assisted extraction (MAE) technique on recovering nutritional and bioactive oils from salmon (Salmo salar) side streams was evaluated and compared to Soxhlet extraction. The response surface methodology (RSM) coupled with a central composite rotatable design was used to optimize time, microwave power, and solid/liquid ratio of the MAE process in terms of oil yield. The optimal MAE conditions were 14.6 min, 291.9 W, 80.1 g/L for backbones, 10.8 min, 50.0 W, 80.0 g/L for heads, and 14.3 min, 960.6 W, 99.5 g/L for viscera, which resulted in a recovery of 69% of the total lipid content for backbones and heads and 92% for viscera. The oils obtained under optimal MAE conditions showed a healthy lipid profile as well as cytotoxic, antioxidant, anti-inflammatory, or antimicrobial properties. These results highlight that oils from underutilized salmon by-products could be exploited by different industrial sectors under the circular economy approach.

Antioxidation, Anti-Inflammation, and Regulation of SRD5A Gene Expression of Oryza sativa cv. Bue Bang 3 CMU Husk and Bran Extracts as Androgenetic Alopecia Molecular Treatment Substances.

Androgenetic alopecia (AGA), a hair loss disorder, is a genetic predisposition to sensitive androgens, inflammation, and oxidative stress. Unfortunately, current treatments with synthetic medicines contain a restricted mechanism along with side effects, whereas the bioactive constituents of plant extracts are multifunctional, with fewer side effects. The massive amounts of rice husk and bran are agricultural wastes that may cause pollution and environmental problems. Owing to these rationales, the local rice variety, Bue Bang 3 CMU (BB3CMU), which is grown in northern Thailand, was evaluated for the valuable utilization of rice by-products, husk (BB3CMU-H) and bran (BB3CMU-RB) extracts, for AGA treatment regarding antioxidant, anti-inflammatory, anti-androgenic activities, and the characterization of bioactive compounds. Our study verified that BB3CMU-H had the highest level of polyphenols, contributing to its greater antioxidant activity. Conversely, BB3CMU-RB was the predominant source of tocopherols, resulting in better anti-androgenic activities regarding the downregulation of steroid 5α-reductase genes (SRD5A). Notably, anti-inflammation via the attenuation of nitric oxide productions was observed in BB3CMU-H (0.06 ± 0.13 µM) and BB3CMU-RB (0.13 ± 0.01 µM), which were significantly comparable to diclofenac sodium salt (0.13 ± 0.19 µM). Therefore, the combination of BB3CMU-H and BB3CMU-RB could be utilized in cosmeceutical and pharmaceutical applications for AGA patients.

High-Throughput Determination of Major Mycotoxins with Human Health Concerns in Urine by LC-Q TOF MS and Its Application to an Exposure Study.

Human biomonitoring constitutes a suitable tool to assess exposure to toxins overcoming the disadvantages of traditional methods. Urine constitutes an accessible biological matrix in biomonitoring studies. Mycotoxins are secondary metabolites produced naturally by filamentous fungi that produce a wide range of adverse health effects. Thus, the determination of urinary mycotoxin levels is a useful tool for assessing the individual exposure to these food contaminants. In this study, a suitable methodology has been developed to evaluate the presence of aflatoxin B2 (AFB2), aflatoxin (AFG2), ochratoxin A (OTA), ochratoxin B (OTB), zearalenone (ZEA), and α-zearalenol (α-ZOL) in urine samples as exposure biomarkers. For this purpose, different extraction procedures, namely, the Solid Phase Extraction (SPE); Dispersive Liquid-Liquid Microextraction (DLLME); and Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) methods were assessed, followed by Liquid Chromatography coupled to Quadrupole Time of Flight Mass Spectrometry with Electrospray Ionization (LC-ESI-QTOF-MS) determination. Then, the proposed methodology was applied to determine mycotoxin concentrations in 56 human urine samples from volunteers and to estimate the potential risk of exposure. The results obtained revealed that 55% of human urine samples analyzed resulted positive for at least one mycotoxin. Among all studied mycotoxins, only AFB2, AFG2, and OTB were detected with incidences of 32, 41, and 9%, respectively, and levels in the range from

High Pressure Processing Impact on Emerging Mycotoxins (ENNA, ENNA1, ENNB, ENNB1) Mitigation in Different Juice and Juice-Milk Matrices.

The aim of the present study was to investigate the potential of high-pressure processing (HPP) (600 MPa during 5 min) on emerging mycotoxins, enniatin A (ENNA), enniatin A1 (ENNA1), enniatin B (ENNB), enniatin B1 (ENNB1) reduction in different juice/milk models, and to compare it with the effect of a traditional thermal treatment (HT) (90 °C during 21 s). For this purpose, different juice models (orange juice, orange juice/milk beverage, strawberry juice, strawberry juice/milk beverage, grape juice and grape juice/milk beverage) were prepared and spiked individually with ENNA, ENNA1, ENNB and ENNB1 at a concentration of 100 µg/L. After HPP and HT treatments, ENNs were extracted from treated samples and controls employing dispersive liquid-liquid microextraction methodology (DLLME) and determined by liquid chromatography coupled to ion-trap tandem mass spectrometry (HPLC-MS/MS-IT). The results obtained revealed higher reduction percentages (11% to 75.4%) when the samples were treated under HPP technology. Thermal treatment allowed reduction percentages varying from 2.6% to 24.3%, at best, being ENNA1 the only enniatin that was reduced in all juice models. In general, no significant differences (p > 0.05) were observed when the reductions obtained for each enniatin were evaluated according to the kind of juice model, so no matrix effects were observed for most cases. HPP technology can constitute an effective tool in mycotoxins removal from juices.

High Efficiency In Vitro Wound Healing of Dictyophora indusiata Extracts via Anti-Inflammatory and Collagen Stimulating (MMP-2 Inhibition) Mechanisms.

Dictyophora indusiata or Phallus indusiatus is widely used as not only traditional medicine, functional foods, but also, skin care agents. Biological activities of the fruiting body from D. indusiata were widely reported, while the studies on the application of immature bamboo mushroom extracts were limited especially in the wound healing effect. Wound healing process composed of 4 stages including hemostasis, inflammation, proliferation, and remodelling. This study divided the egg stage of bamboo mushroom into 3 parts: peel and green mixture (PGW), core (CW), and whole mushroom (WW). Then, aqueous extracts were investigated for their nucleotide sequencing, biological compound contents, and wound healing effect. The anti-inflammatory determination via the levels of cytokine releasing from macrophages, and the collagen stimulation activity on fibroblasts by matrix metalloproteinase-2 (MMP-2) inhibitory activity were determined to serve for the wound healing process promotion in the stage 2-4 (wound inflammation, proliferation, and remodelling of the skin). All D. indusiata extracts showed good antioxidant potential, significantly anti-inflammatory activity in the decreasing of the nitric oxide (NO), interleukin-1 (IL-1), interleukin-1 (IL-6), and tumour necrosis factor-α (TNF-α) secretion from macrophage cells (p < 0.05), and the effective collagen stimulation via MMP-2 inhibition. In particular, CW extract containing high content of catechin (68.761 ± 0.010 mg/g extract) which could significantly suppress NO secretion (0.06 ± 0.02 µmol/L) better than the standard anti-inflammatory drug diclofenac (0.12 ± 0.02 µmol/L) and their MMP-2 inhibition (41.33 ± 9.44%) was comparable to L-ascorbic acid (50.65 ± 2.53%). These findings support that CW of D. indusiata could be an essential natural active ingredient for skin wound healing pharmaceutical products.

Impact of Pressurized Liquid Extraction and pH on Protein Yield, Changes in Molecular Size Distribution and Antioxidant Compounds Recovery from Spirulina.

The research aims to extract nutrients and bioactive compounds from spirulina using a non-toxic, environmentally friendly and efficient method-Pressurized Liquid Extraction (PLE). In this work, Response Surface Methodology (RSM)-Central Composite Design (CCD) was used to evaluate and optimize the extraction time (5-15 min), temperature (20-60 °C) and pH (4-10) during PLE extraction (103.4 bars). The multi-factor optimization results of the RSM-CCD showed that under the pressure of 103.4 bars, the optimal conditions to recover the highest content of bioactive compounds were 10 min, 40 °C and pH 4. Furthermore, the compounds and antioxidant capacity of PLE and non-pressurized extraction extracts were compared. The results showed that under the optimal extraction conditions (10 min, 40 °C and pH 4), PLE significantly improved the antioxidant capacity (2870.5 ± 153.6 µM TE), protein yield (46.8 ± 3.1%), chlorophyll a (1.46 ± 0.04 mg/g), carotenoids (0.12 ± 0.01 mg/g), total polyphenols (11.49 ± 0.04 mg/g) and carbohydrates content (78.42 ± 1.40 mg/g) of the extracts compared with non-pressurized extraction (p < 0.05). The protein molecular distribution of the extracts was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the results showed that there were more small-molecule proteins in PLE extracts. Moreover, Liquid Chromatography Triple Time of Flight Mass Spectrometry (TOF-LC-MS-MS) was used to analyze the phenolic profile of the extracts, and the results showed the extracts were rich on phenolic compounds, such as p-coumaric acid and cinnamic acid being the predominant phenolic compounds in the PLE extract. This indicates that PLE can promote the extraction of bioactive compounds from Spirulina, which is of great significance for the application of PLE technology to obtain active substances from marine algae resources.

Assessment of Human Exposure to Deoxynivalenol, Ochratoxin A, Zearalenone and Their Metabolites Biomarker in Urine Samples Using LC-ESI-qTOF.

Human are exposed to a wide range of mycotoxins through dietary food intake, including processed food. Even most of the mycotoxin exposure assessment studies are based on analysis of foodstuffs, and evaluation of dietary intake through food consumption patterns and human biomonitoring methods are rising as a reliable alternative to approach the individual exposures, overcoming the limitations of the indirect dietary assessment. In this study, human urine samples were analyzed, seeking the presence of deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZEA), and their metabolites. For this purpose, 40 urine samples from female and male adult residents in the city of Valencia (Spain) were evaluated by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-ESI-qTOF) after salting-out liquid-liquid extraction. Analytical data showed that 72.5% of analyzed samples were contaminated by at least one mycotoxin at variable levels. The most prevalent mycotoxins were de-epoxy DON (DOM-1) (53%), ZEA (40%), and α-zearalenol (αZOL) (43%), while OTA was only detected in one sample. The mean concentrations in positive samples were DON (9.07 ng/mL), DOM-1 (20.28 ng/mL), ZEA (6.70 ng/mL), ZEA-14 glucoside (ZEA-14-Glc) (12.43 ng/mL), αZOL (27.44 ng/mL), αZOL-14 glucoside (αZOL-14-Glc) (12.84 ng/mL), and OTA (11.73 ng/mL). Finally, probable daily intakes (PDIs) were calculated and compared with the established tolerable daily intakes (TDIs) to estimate the potential risk of exposure to the studied mycotoxins. The calculated PDI was below the TDI value established for DON in both female and male adults, reaching a percentage up to 30%; however, this percentage increased up to 92% considering total DON (DON + DOM-1). On the other hand, the PDI obtained for ZEA and its metabolites were higher than the TDI value fixed, but the low urine excretion rate (10%) considered should be highlighted. Finally, the PDI calculated in the detected positive sample for OTA exceeded the TDI value. The findings of the present study confirm the presence of the studied mycotoxins and their metabolites as some of the most prevalent in urine.

Salmon (Salmo salar) Side Streams as a Bioresource to Obtain Potential Antioxidant Peptides after Applying Pressurized Liquid Extraction (PLE).

The pressurized liquid extraction (PLE) technique was used to obtain protein extracts with antioxidant capacity from salmon muscle remains, heads, viscera, skin, and tailfins. A protein recovery percentage ≈28% was obtained for all samples except for viscera, which was ≈92%. These values represented an increase of 1.5-4.8-fold compared to stirring extraction (control). Different SDS-PAGE profiles in control and PLE extracts revealed that extraction conditions affected the protein molecular weight distribution of the obtained extracts. Both TEAC (Trolox equivalent antioxidant capacity) and ORAC (oxygen radical antioxidant capacity) assays showed an outstanding antioxidant activity for viscera PLE extract. Through liquid chromatography coupled with electrospray ionization triple time-of-flight (nanoESI qQTOF) mass spectrometry, 137 and 67 peptides were identified in control and PLE extracts from salmon viscera, respectively None of these peptides was found among the antioxidant peptides inputted in the BIOPEP-UMP database. However, bioinformatics analysis showed several antioxidant small peptides encrypted in amino acid sequences of viscera extracts, especially GPP (glycine-proline-proline) and GAA (glycine-alanine-alanine) for PLE extracts. Further research on the relationship between antioxidant activity and specific peptides from salmon viscera PLE extracts is required. In addition, the salmon side streams studied presented non-toxic levels of As, Hg, Cd, and Pb, as well as the absence of mycotoxins or related metabolites. Overall, these results confirm the feasible use of farmed salmon processing side streams as alternative sources of protein and bioactive compounds for human consumption.

Dietary Exposure to Mycotoxins through Alcoholic and Non-Alcoholic Beverages in Valencia, Spain.

The present study investigated the presence of 30 mycotoxins in 110 beverage samples of beer, wine, cava, and cider purchased in Valencia (Spain). A validated method based on dispersive liquid-liquid microextraction and chromatographic methods coupled with tandem mass spectrometry was applied. The method showed satisfactory recoveries ranging from 61 to 116% for the different beverages studied. The detection and quantification limits ranged from 0.03 to 2.34 µg/L and 0.1 to 7.81 µg/L, respectively. The results showed that beer samples were the most contaminated, even with concentrations ranging from 0.24 to 54.76 µg/L. A significant presence of alternariol was found in wine, which reached concentrations up to 26.86 µg/L. Patulin and ochratoxin A were the most frequently detected mycotoxins in cava and cider samples, with incidences of 40% and 26%, respectively. Ochratoxin A exceeded the maximum level set by the EU in one wine sample. The results obtained were statistically validated. The combined exposure was assessed by the sum of mycotoxin concentrations contaminating the same samples to provide information on the extent of dietary exposure to mycotoxins. No significant health risk to consumers was associated with the mycotoxin levels detected in the beverages tested.

High Pressure Processing Impact on Alternariol and Aflatoxins of Grape Juice and Fruit Juice-Milk Based Beverages.

High-pressure processing (HPP) has emerged over the last 2 decades as a good alternative to traditional thermal treatment for food safety and shelf-life extension, supplying foods with similar characteristics to those of fresh products. Currently, HPP has also been proposed as a useful tool to reduce food contaminants, such as pesticides and mycotoxins. The aim of the present study is to explore the effect of HPP technology at 600 MPa during 5 min at room temperature on alternariol (AOH) and aflatoxin B1 (AFB1) mycotoxins reduction in different juice models. The effect of HPP has also been compared with a thermal treatment performed at 90 °C during 21 s. For this, different juice models, orange juice/milk beverage, strawberry juice/milk beverage and grape juice, were prepared and spiked individually with AOH and AFB1 at a concentration of 100 µg/L. After HPP and thermal treatments, mycotoxins were extracted from treated samples and controls by dispersive liquid-liquid microextraction (DLLME) and determined by HPLC-MS/MS-IT. The results obtained revealed reduction percentages up to 24% for AFB1 and 37% for AOH. Comparing between different juice models, significant differences were observed for AFB1 residues in orange juice/milk versus strawberry juice/milk beverages after HPP treatment. Moreover, HPP resulted as more effective than thermal treatment, being an effective tool to incorporate to food industry in order to reach mycotoxins reductions.

Biomonitoring of Multiple Mycotoxins in Urine by GC-MS/MS: A Pilot Study on Patients with Esophageal Cancer in Golestan Province, Northeastern Iran.

A pilot study to investigate the occurrence of 10 mycotoxins (deoxynivalenol, DON; 3-acetyldeoxynivalenol, 3-ADON; 15-acetyldeoxynivalenol, 15-ADON; fusarenon-X, FUS-X; diacetoxyscirpenol, DAS; nivalenol, NIV; neosolaniol, NEO; zearalenone, ZON; zearalanone, ZAN; T-2 toxin, T-2; and HT-2 toxin, HT-2) in esophageal cancer patients was performed with the urinary biomarkers approach in Golestan, Iran. Urine multimycotoxin analysis was performed by dispersive liquid-liquid microextraction and gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis, and values were normalized with urinary creatinine (µg/g). Four mycotoxins, namely NEO (40%), HT-2 (17.6%), DON (10%), and HT-2 (5.8%), were detected in the analyzed urine samples. DON was only detected in the control group (5.09 µg/g creatinine), while T-2 (44.70 µg/g creatinine) was only present in the esophageal cancer group. NEO and HT-2 were quantified in both control and case groups, showing average of positive samples of 9.09 and 10.45 µg/g creatinine for NEO and 16.81 and 29.09 µg/g creatinine for HT-2, respectively. Mycotoxin co-occurrence was observed in three samples as binary (NEO/HT-2 and T-2/HT-2) and ternary (DON/NEO/HT-2) combinations, reaching total concentrations of 44.58, 79.13, and 30.04 µg/g creatinine, respectively. Further investigations are needed to explore a causal association between mycotoxin contamination and esophageal cancer. For this pilot study in Golestan, the low sample size was a very limiting factor.

Development of Antioxidant Protein Extracts from Gilthead Sea Bream (Sparus aurata) Side Streams Assisted by Pressurized Liquid Extraction (PLE).

The pressurized liquid extraction (PLE) technique was used, for the first time, to obtain protein extracts with antioxidant activity from side streams (muscle, heads, viscera, skin, and tailfins) of gilthead sea bream (Sparus aurata) in order to give added value to these underutilized matrices. Extraction conditions previously optimized for sea bass (Dicentrarchus labrax) side streams were applied. Protein recovery percentages were 22% (muscle), 33% (heads), 78% (viscera), 24% (skin), and 26% (tailfins), which represented an increase of 1.2-4.5-fold compared to control samples (extraction by stirring). The SDS-PAGE profiles revealed that PLE-assisted extraction influenced protein molecular weight distribution of the obtained extracts. PLE conditions also allowed increasing the antioxidant capacity measured by both Trolox equivalent antioxidant capacity (TEAC; 1.3-2.4 fold) and oxygen radical absorbance capacity (ORAC; 1.9-6.4) assays for all fish extracts. Inductively coupled plasma mass spectrometry (ICP-MS) and high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-qTOF-MS) were used to investigate the presence of toxic metals and mycotoxins in sea bream side streams. The levels of As, Hg, Cd, and Pb were below those established by authorities for fish muscle for human consumption (except for Cd in viscera samples). Through a nontargeted screening approach, no mycotoxins or related metabolites were detected for all sea bream side streams. This study contributes to the research on the valorization of fish processing side streams using environmentally friendly technology.