Comparative assessment of three extraction procedures for determination of emerging Fusarium mycotoxins in pasta by LC–MS/MS

Abstract A new rapid, sensitive, reproducible and reliable method was developed for the quantitative determination of enniatins A, A1, B and B1, beauvericin and fusaproliferin in dry and fresh pasta by liquid chromatography-triple quadrupole-tandem mass spectrometry. A comparative study of different rapid and economical extraction procedures was performed for the extraction of these mycotoxins in pasta. For this purpose, three different approaches were studied during the extraction step (Ultra-Turrax, ultrasonic bath and microwave). Optimal extraction conditions were reached using Ultra-Turrax with acetonitrile for 3 min without purification step. The chromatographic separation of the six m…

Evolution of emerging Fusarium mycotoxins contents throughout the shelf-life period of food

which is cooked by steaming. It has been historically eaten in North African countries but nowadays its consumption is widely extended all around the world. As a cereal-based food, semolina is highly susceptible to contamination by mycotoxin-producing fungi. The presented procedure involves a modified QuEChERSbased extraction of 24 mycotoxins (15-ADON, 3-ADON, AFLAB1, AFLAB2, AFLAG1, AFLAG2, BEA, DON, DAS, ENA, ENA1, ENB, ENB1, FB1, FB2, FB3, FUSX, HT-2, NEO, NIV, OTA, STG, T2, ZEA) produced by Aspergillus, Penicillum and Fusarium fungi. The validation was performed by analyzing recovery samples at three different spiked concentrations with four replicates (n=4) at each concentration. Recov…

Risk assessment of beauvericin, enniatins and fusaproliferin present in follow-up infant formula by in vitro evaluation of the duodenal and colonic bioaccessibility

Abstract In this study, 72 samples of follow-up infant formula of Spanish origin were analyzed for the presence of the mycotoxins beauvericin (BEA), enniatins (ENs) (A, A1, B, B1), and fusaproliferin (FUS). The samples analyzed were extracted three times with ethyl acetate and then the mycotoxins were identified and quantified using a liquid chromatography (LC) coupled to a diode array detector (DAD). The positive samples analyzed in this study were digested through a simulated gastrointestinal digestion model, which permit to simulate the physiological condition of the human gastrointestinal tract (duodenal and colonic compartments) in order to assess the bioaccessibility of the bioactive …

First report on the presence of emerging Fusarium mycotoxins enniatins (A, A1, B, B1), beauvericin and fusaproliferin in rice on the Moroccan retail markets

Seventy samples of rice purchased from local markets in six cities from Morocco (Rabat, Casablanca, Kenitra, Mohammadia, Tanger and Errachidia) were analyzed for the presence of six emerging mycotoxins: four enniatins ENs (ENA, ENA1, ENB and ENB1), beauvericin (BEA) and fusaproliferin (FUS). Samples were extracted with a mixture of acetonitrile/water (85/15, v/v) by using an ultra-turrax homogenizer. Mycotoxins were then identified and quantified with liquid chromatography (LC) coupled to diode array detector (DAD). Positive samples were confirmed with an LC-MS/MS. Analytical results showed that BEA was present in 75.7% of total analyzed samples. BEA levels varied between 3.8 and 26.3 mg/kg…

A preliminary study in Wistar rats with enniatin : A contaminated feed

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 two months-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 twenty-eight 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, duode…

Emerging Fusarium mycotoxins in organic and conventional pasta collected in Spain

Abstract 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 pos…

Influence of the making and cooking pasta on enniatins contents

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 …

Development a mitigation strategy of enniatins in pasta under home-cooking conditions

The effect of pasta cooking on the enniatins (ENs) contents was evaluated in white and whole-grain pasta. The cooking of pasta was performed for 10 min in boiling water. ENs were extracted from raw and cooked pasta by Solid-Liquid extraction with acetonitrile, and from cooking water by Dispersive Liquid–Liquid Micro-Extraction (DLLME) with a mixture of acetonitrile and carbone tetrachloride. Mycotoxin analysis was performed by Liquid Chromatography coupled to Tandem Mass Spectrometry. The effect of pasta cooking has a marked influence on the reduction of ENs levels. Reductions of 98–100% for ENA, 94–95% for ENA1, 14–49% for ENB and 53–65% for ENB1 were achieved after the process. ENs were d…

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…

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%…