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.

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.

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.

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.

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 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.

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.

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.

Aquaculture and its by-products as a source of nutrients and bioactive compounds.

Underutilized marine resources (e.g., algae, fish, and shellfish processing by-products), as sustainable alternatives to livestock protein and interesting sources of bioactive compounds, have attracted the attention of the researchers. Aquatic products processing industries are growing globally and producing huge amounts of by-products that often discarded as waste. However, recent studies pointed out that marine waste contains several valuable components including high-quality proteins, lipids, minerals, vitamins, enzymes, and bioactive compounds that can be used against cancer and some cardiovascular disorders. Besides, previously conducted studies on algae have shown the presence of some unique biologically active compounds and valuable proteins. Hence, this chapter points out recent advances in this area of research and discusses the importance of aquaculture and fish processing by-products as alternative sources of proteins and bioactive compounds.

Scaling-up processes: Patents and commercial applications.

There is currently a great demand for fish and seafood products. However, their high consumption produces large quantities of by-products that can be an ecological problem. That is why it is necessary to look for alternatives to revalue these products and give them a second life, thus reducing their environmental impact. In this sense, several investigations have been carried out in laboratories around the world to extract compounds from marine processing industry for the final high added-value products. Some of these compounds are collagen, omega 3 fatty acids, protein concentrates or chitin/chitosan, among others. Nevertheless, one of the critical steps for obtaining these compounds at the industrial level is the scale-up. Much of this research does not progress at the industrial level due to the complications of its large-scale research and use. However, the advances in technology entail that more research studies achieve to reach the industrial development phase. Once a product or process has been developed, it should be patented to protect its intellectual property. This chapter gives an overview of this entire process, showing some examples of patents or products from seafood by-products already marketed and providing some details of the corresponding legislation.

Development and Validation of a LC-ESI-MS/MS Method for the Determination of Alternaria Toxins Alternariol, Alternariol Methyl-Ether and Tentoxin in Tomato and Tomato-Based Products.

Alternaria species are capable of producing several secondary toxic metabolites in infected plants and in agricultural commodities, which play important roles in food safety. Alternaria alternata turn out to be the most frequent fungal species invading tomatoes. Alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN) are some of the main Alternaria mycotoxins that can be found as contaminants in food. In this work, an analytical method based on liquid chromatography (LC) tandem mass spectrometry (MS/MS) detection for the simultaneous quantification of AOH, AME, and TEN in tomato and tomato-based products was developed. Mycotoxin analysis was performed by dispersive liquid-liquid microextraction (DLLME) combined with LC-ESI-MS/MS. Careful optimization of the MS/MS parameters was performed with an LC/MS system with the ESI interface in the positive ion mode. Mycotoxins were efficiently extracted from sample extract into a droplet of chloroform (100 µL) by DLLME technique using acetonitrile as a disperser solvent. Method validation following the Commission Decision No. 2002/657/EC was carried out by using tomato juice as a blank matrix. Limits of detection and quantitation were, respectively, in the range 0.7 and 3.5 ng/g. Recovery rates were above 80%. Relative standard deviations of repeatability (RSDr) and intermediate reproducibility (RSDR) were ≤ 9% and ≤ 15%, respectively, at levels of 25 and 50 ng/g. Five out of 30 analyzed samples resulted positive to at least one Alternaria toxin investigated. AOH was the most common Alternaria toxin found, but at levels close to LOQ (average content: 3.75 ng/g).

Analysis of trichothecenes in laboratory rat feed by gas chromatography-tandem mass spectrometry.

A method for the determination of seven trichothecenes, neosolaniol (NEO), diacetoxyscirpenol (DAS), deoxynivalenol (DON), nivalenol (NIV), fusarenon-X (FUS-X), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON), in laboratory rat feed by GC-MS/MS was developed. Sample extraction and purification was performed by an acidified mixture of acetonitrile/water (80-20% v/v). Limits of quantitation (LOQs) were between 1 and 10 µg kg(-1) for all studied trichothecenes. Eight concentration levels between the LOQ and 100 × LOQ were used for the calibration curves. Matrix-matched calibration was used for quantitation purposes to compensate the detector signal enhancement obtained for all the analytes. The method accuracy was evaluated by recovery assays at three concentration levels, 25, 50 and 100 µg kg(-1) (n = 9). Recoveries ranged from 62% to 97% and precision, expressed as intra- and inter-day relative standard deviations, was evaluated for all compounds. The validated method was successfully applied to the analysis of 35 laboratory rat feed samples showing mycotoxin contamination in 66% of the samples. DON was the most prevalent trichothecene followed by 15-ADON, NIV and 3-ADON. The maximum DON concentration reached in real samples was 2156 ± 4.3 µg kg(-1), while NEO, DAS and FUS-X were not detected in any sample. Multi-contamination by at least two mycotoxins was observed in 17% of the analysed feed samples.

Quantitative determination of trichothecenes in breadsticks by gas chromatography-triple quadrupole tandem mass spectrometry.

Breadsticks are pencil-sized sticks of dry bread widely consumed as a pre-meal appetiser. They are basically wheat-based snacks, which makes them a good matrix to evaluate mycotoxin contamination, since wheat is very susceptible to fungal attack. In this sense, the fast, selective and sensitive gas chromatography-triple quadrupole tandem mass spectrometry (GC-QqQ-MS/MS) method proposed here allows simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol, fusarenon-X, diacetoxyscirpenol, nivalenol, neosolaniol, HT-2 and T-2 toxin in breadsticks after QuEChERS extraction and clean-up. The performance of the method was assessed with respect to European Commission Regulations by studying the selectivity and specificity, limit of detection (LOD), limit of quantitation (LOQ), linearity, matrix effect, accuracy, precision and trueness. Satisfactory results in terms of validation parameters were obtained for all selected mycotoxins (recovery range of 70-110%, RSD < 10%, LOQ <40 µg kg(-1)). The trueness of the method was supported by using certified reference material (DON 1062 ± 110 µg kg(-1)). The method was successfully used to evaluate the occurrence of the studied Fusarium toxins in 61 breadstick samples. A total of 64% of the samples showed mycotoxin contamination, DON being the most frequently detected toxin. Nonetheless, mean levels obtained were far below the maximum levels permitted by European Union legislation. An additional goal was to carry out a risk-characterisation approach to DON by comparing probable daily intake and provisional maximum tolerable daily intake (PMTDI).

Determination of mycotoxins in bee pollen by gas chromatography-tandem mass spectrometry.

Bee pollen, promoted as a natural food supplement, is consumed increasingly by people to maintain a healthy diet. Depending on environmental conditions, pollen can also be an optimum medium for growth of molds such as Fusarium and Penicillium . A quick, easy, cheap, rapid, and safe (QuEChERS) extraction procedure followed by a gas chromatography-tandem mass spectrometry (GC-MS/MS) determination of eight selected Fusarium toxins in bee pollen was developed and optimized. Recovery studies at 20, 80, and 1000 µg/kg showed values between 73 and 95% with relative standard deviations (RSDs) of <15% for all studied mycotoxins. Limits of quantitation (LOQ) ranged from 1 to 4 µg/kg. The proposed method was applied to the analysis of 15 commercial samples. Two of 15 samples showed quantifiable values for neosolaniol and nivalenol.

Multi-mycotoxin analysis in wheat semolina using an acetonitrile-based extraction procedure and gas chromatography-tandem mass spectrometry.

A new analytical method for the rapid and simultaneous determination of ten mycotoxins including patulin, zearalenone and eight trichothecenes (nivalenol, fusarenon-X, diacetoxyscirpenol, 3-acetyl-deoxynivalenol, neosolaniol, deoxynivalenol, T-2 and HT-2) in wheat semolina has been developed and optimized. Sample extraction and purification were performed with a modified QuEChERS-based (acronym of Quick, Easy, Cheap, Effective, Rugged and Safe) procedure and determined by gas chromatography (GC) coupled to triple quadrupole instrument (QqQ). This is the first paper on the application of GC-QqQ-MS/MS to analysis of mycotoxins. Careful optimization of the gas chromatography-tandem mass spectrometry parameters was achieved in order to attain a fast separation with the best sensitivity allowing a total run time of 16 min. The validation was performed by analyzing recovery samples at three different spiked concentrations, 20, 40 and 80 µg kg(-1), with four replicates (n=4) at each concentration. Recoveries ranged from 74% to 124% and the intra-day precision and inter-day precision, expressed as relative standard deviation, were lower than 13% and 17%, respectively for all studied compounds, except for zearalenone. Limits of quantification (LOQ) were lower than 10 µg kg(-1) for all studied mycotoxins. Eight concentration levels were used for constructing the calibration curves which showed good linearity between LOQ and 100 times LOQ concentration levels (linear range). Matrix-matched calibration for applying the method in routine analysis is recommended for reliable quantitative results. The method validated was successfully applied to fifteen wheat semolina samples detecting occurrence of mycotoxins at concentrations below the maximum permissible level.