A.B. Serrano

Co-occurrence and risk assessment of mycotoxins in food and diet from Mediterranean area.

The contents of 14 mycotoxins were studied in samples of different cereals and cereal products from four countries of the Mediterranean region. Two hundred and sixty-five samples from Spain, Italy, Morocco and Tunisia were analysed. Samples were extracted with matrix solid-phase dispersion (MSPD) and determined by liquid chromatography-tandem mass spectrometry with a triple quadrupole mass analyser. The percentage of total samples contaminated was 53%. The frequency of contaminated samples from Spain, Italy, Tunisia and Morocco was 33%, 52%, 96% and 50%, respectively. Nivalenol and beauvericin were the most predominant mycotoxins. This is the first international report to study the presence of several mycotoxins in different types of cereal (rice, wheat, maize, rye, barley, oat, spelt and sorghum) and cereal products (snacks, pasta, soup, biscuits and flour) from the Mediterranean area, estimate the intake of mycotoxins and evaluate the risk assessment.

Emerging Fusarium mycotoxins in organic and conventional pasta collected in Spain.

One of the main sources of emerging Fusarium mycotoxins in human nutrition is the cereals and cereal products. In this study, an analytical method to determine enniatins A, A1, B and B1 (ENs), beauvericin (BEA) and fusaproliferin (FUS) based on Ultra-Turrax extraction followed by liquid chromatography coupled to triple quadrupole mass spectrometer detector (MS/MS QqQ), was applied for the analysis of pasta. For this purpose, 114 commercial samples of pasta were acquired from supermarkets located in Valencia. The results showed higher frequencies of contamination in organic pasta than in conventional pasta, while the concentration levels were variable for both types of pasta. In positive samples, BEA levels varied from 0.10 to 20.96μg/kg and FUS levels varied from 0.05 to 8.02μg/kg. ENs levels ranged from 0.25 to 979.56μg/kg, though the majority of the values were below 25μg/kg. Besides, it was observed the simultaneous presence of two or more mycotoxins in a high percentage of the samples. Finally, an evaluation of the dietary exposure of the emerging Fusarium mycotoxins was performed in the Spanish population. The prevalence of ENs, BEA and FUS in cereal products suggests that the toxins may pose a health risk to Spanish population.

A preliminary study in Wistar rats with enniatin A contaminated feed

Abstract A 28-day repeated dose preliminary assay, using enniatin A naturally contaminated feed through microbial fermentation by a Fusarium tricinctum strain, was carried out employing 2-month-old female Wistar rats as in vivo experimental model. In order to simulate a physiological test of a toxic compound naturally produced by fungi, five treated animals were fed during 28 days with fermented feed. As control group, five rats were fed with standard feed. At the 28th day, blood samples were collected for biochemical analysis and the gastrointestinal tract, liver and kidneys were removed from each rat for enniatin A detection and quantitation. Digesta were collected from stomach, duodenum, jejunum, ileum and colon. Enniatin A present in organs and in biological fluids was analyzed by liquid chromatography-diode array detector (LC-DAD) and confirmed by LC-mass spectrometry linear ion trap (MS-LIT); also several serum biochemical parameters and a histological analysis of the duodenal tract were performed. No adverse effects were found in any treated rat at the enniatin A concentration (20.91 mg/kg bw/day) tested during the 28-day experiment. Enniatin A quantitation in biological fluids ranged from 1.50 to 9.00 mg/kg, whereas in the gastrointestinal organs the enniatin A concentration ranged from 2.50 to 23.00 mg/kg. The high enniatin A concentration found in jejunum liquid and tissue points to them as an absorption area. Finally, two enniatin A degradation products were identified in duodenum, jejunum and colon content, probably produced by gut microflora.

Development of a Rapid LC-MS/MS Method for the Determination of Emerging Fusarium mycotoxins Enniatins and Beauvericin in Human Biological Fluids

A novel method for the simultaneous determination of enniatins A, A1, B and B1 and beauvericin, both in human urine and plasma samples, was developed and validated. The method consisted of a simple and easy pretreatment, specific for each matrix, followed by solid phase extraction (SPE) and detection by high performance liquid chromatography-tandem mass spectrometry with an electrospray ion source. The optimized SPE method was performed on graphitized carbon black cartridges after suitable dilution of the extracts, which allowed high mycotoxin absolute recoveries (76%–103%) and the removal of the major interferences from the matrix. The method was extensively evaluated for plasma and urine samples separately, providing satisfactory results in terms of linearity (R2 of 0.991–0.999), process efficiency (>81%), trueness (recoveries between 85% and 120%), intra-day precision (relative standard deviation, RSD < 18%), inter-day precision (RSD < 21%) and method quantification limits (ranging between 20 ng·L−1 and 40 ng·L−1 in plasma and between 5 ng·L−1 and 20 ng·L−1 in urine). Finally, the highly sensitive validated method was applied to some urine and plasma samples from different donors.

Degradation study of enniatins by liquid chromatography-triple quadrupole linear ion trap mass spectrometry.

Enniatins A, A1, B and B1 (ENs) are mycotoxins produced by Fusarium spp. and are normal contaminants of cereals and derivate products. In this study, the stability of ENs was evaluated during food processing by simulation of pasta cooking. Thermal treatments at different incubation times (5, 10 and 15 min) and different pH (4, 7 and 10) were applied in an aqueous system and pasta resembling system (PRS). The concentrations of the targeted mycotoxins were determined using liquid chromatography coupled to tandem mass spectrometry. High percentages of ENs reduction (81-100%) were evidenced in the PRS after the treatments at 5, 10 and 15 min of incubation. In contrast to the PRS, an important reduction of the ENs was obtained in the aqueous system after 15 min of incubation (82-100%). In general, no significant differences were observed between acid, neutral and basic solutions. Finally, several ENs degradation products were identified using the technique of liquid chromatography-triple quadrupole linear ion trap mass spectrometry.

Effects of technological processes on enniatin levels in pasta.

BACKGROUND Potential human health risks posed by enniatins (ENs) require their control primarily from cereal products, creating a demand for harvesting, food processing and storage techniques capable to prevent, reduce and/or eliminate the contamination. In this study, different methodologies to pasta processing simulating traditional and industrial processes were developed in order to know the fate of the mycotoxin ENs. The levels of ENs were studied at different steps of pasta processing. The effect of the temperature during processing was evaluated in two types of pasta (white and whole-grain pasta). Mycotoxin analysis was performed by LC-MS/MS. RESULTS High reductions (up to 50% and 80%) were achieved during drying pasta at 45-55°C and 70-90°C, respectively. The treatments at low temperature (25°C) did not change EN levels. The effect of pasta composition did not cause a significant effect on the stability of ENs. The effect of the temperature allowed a marked mycotoxin reduction during pasta processing. Generally, ENA1 and ENB showed higher thermal stability than did ENA and ENB1 . CONCLUSIONS The findings from the present study suggested that pasta processing at medium-high temperatures is a potential tool to remove an important fraction of ENs from the initial durum wheat semolina.