Application of hybrid linear ion trap-high resolution mass spectrometry to the analysis of mycotoxins in beer

This paper reports the application of liquid chromatography electrospray ionization ion trap-orbitrap mass spectrometry for the determination of 18 mycotoxins (aflatoxins, fumonisins, trichothecenes, ochratoxin A, sterogmatocystin, beauvaricin, zearalenone and zearalenol) in beer. The extraction procedure was carried out by solid phase extraction (SPE): SPE columns were conditioned with acetonitrile/methanol and water. Beer was loaded onto the column which was washed with water. In these conditions, the recoveries were more than 65% and the relative standard deviation (RSD) were below 18%. The lowest limits of quantification (LLOQ) ranged from 9 to 155 ng ml(-1). Matrix-matched calibration …

Occurrence of fourteen mycotoxins in tiger-nuts

Abstract A previous developed matrix solid-phase dispersion (MSPD) extraction method was applied for the routine analysis of aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FB 1 and FB 2 ), beauvericin (BEA), nivalenol (NIV), deoxynivalenol (DON), the toxin T-2 (T-2), toxin HT-2 (HT-2), diacetoxyscirpenol (DAS) and zearalenone (ZEN) in tiger-nuts by liquid chromatography–triple-quadrupole linear ion trap (HPLC–QTRAP ® ). The extraction solid support used was C 18 , while the elution solvent was acetonitrile/methanol (50/50, v/v) 1 mM ammonium formate. Using matrix-matched calibration, recoveries and repeatabilities were in the range 67–89% and 2–11% relative standard deviation (RSD), resp…

Applicability of hybrid linear ion trap-high resolution mass spectrometry and quadrupole-linear ion trap-mass spectrometry for mycotoxin analysis in baby food.

Abstract Recent developments in mass spectrometers have created a paradoxical situation; different mass spectrometers are available, each of them with their specific strengths and drawbacks. Hybrid instruments try to unify several advantages in one instrument. In this study two of wide-used hybrid instruments were compared: hybrid quadrupole-linear ion trap-mass spectrometry (QTRAP ® ) and the hybrid linear ion trap-high resolution mass spectrometry (LTQ-Orbitrap ® ). Both instruments were applied to detect the presence of 18 selected mycotoxins in baby food. Analytical parameters were validated according to 2002/657/CE. Limits of quantification (LOQs) obtained by QTRAP ® instrument ranged …

Determination of mycotoxins in multicereal flour by matrix solid phase dispersion and LC–MS/MS

Survey of microbial quality of plant-based foods served in restaurants

This study was carried out to evaluate the microbiological quality of plant-based foods obtained from foodservice establishments. The samples included cereals, legumes, fruits and vegetables. According to the European Commission Regulation (No. 2073/2005 and No. 1441/2007) and Spanish microbiological criteria (No. 3484/2000), vegetables were the plant-based dishes where more samples exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae. Furthermore, Staphylococcus aureus and Escherichia coli were also found in several vegetable dishes. E. coli and Salmonella spp. were detected in 6.6% and 0.7% of lettuce samples, respectively. However, all the samples were negative f…

Incidence of microorganisms from fresh orange juice processed by squeezing machines

Abstract This study was carried out to evaluate the microbiological quality of orange juice obtained from squeezing machines in foodservice establishments. The samples included fresh squeezed orange juice and juice which is maintained in metal jugs until consumption. According to the European Commission Regulation (No. 2073/2005 and No. 1441/2007) and Spanish microbiological criteria (No. 3484/2000), 12% and 43% of the total examined lots exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae, respectively. Possibly, this contamination is caused by incorrect handling of oranges and juices and also by inadequate cleaning and sanitization of squeezing machine and metal …

Metabolomic Changes after Coffee Consumption: New Paths on the Block

Scope Several studies suggest that regular coffee consumption may help preventing chronic diseases, but the impact of daily intake and the contribution of coffee metabolites in disease prevention are still unclear. The present study aimed at evaluating whether and how different patterns of coffee intake (one cup of espresso coffee/day, three cups of espresso coffee/day, one cup of espresso coffee/day and two cocoa-based products containing coffee two times per day) might impact endogenous molecular pathways. Methods and results A three-arm, randomized, cross-over trial was performed in 21 healthy volunteers who consumed each treatment for one month. Urine samples were collected to perform u…

Application of an HPLC-MS/MS method for mycotoxin analysis in commercial baby foods

Abstract This article describes the validation of an analytical method for the detection of 21 mycotoxins in baby food. The analytical method is based on the simultaneous extraction of selected mycotoxins by matrix solid-phase dispersion (MSPD) followed by liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) using a hybrid triple quadrupole-linear ion trap mass spectrometer (QTRAP®). Information Dependent Acquisition (IDA), an extra confirmation tool for samples that contain the selected mycotoxins, was used. The matrix effects were evaluated, and the corrections for the matrix effects were performed using two calibration approaches: external matrix-matched calibration and…

Glucose influence on the production of T-2 toxin by Fusarium sporotrichioides

Toxigenic isolate of Fusarium sporotrichioides was tested for the T-2 toxin production on PDA plates during 10 days under various glucose concentrations. T-2 toxin was determined by LC-MS and confirmed with LC-MS/MS. This analytical method has been applied, for the first time, to an extensive study of T-2 accumulation. Results showed that the production of this mycotoxin is directly correlated to the concentration of glucose present in the medium. Concentrations of T-2 toxin produced by the strain of F. sporotrichioides ranged from 0 to 1.45 mg/kg. The better T-2 production was evidenced in the fermentation operated with 20% of glucose.

Occurrence of fumonisins in organic and conventional cereal-based products commercialized in France, Germany and Spain

A fast and effective method for quantitative determination of fumonisins B1 and B2 (FB1 and FB2) in different cereal-based products was developed. Fumonisins were extracted combining a solid liquid extraction (SLE) pre-treatment and immunoaffinity columns (IACs), which were applied for an effective clean-up. Consecutively, high pressure liquid chromatography (HPLC) coupled to hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap®) was used for the separation, detection and quantification of targeted mycotoxins. The analytical method showed acceptable recoveries ranged from 89.7% to 99.1%, as well as a relative standard deviation (%, RSD) lower than 12%. A total of 1250 samples f…

Microbial contamination of milk and dairy products from restaurants in Spain

This study was carried out to evaluate the microbiological quality of milk and some dairy products, including 95 lots of warm milk, 95 lots of milk conserved at room temperature, and 75 lots of dairy products collected from restaurants in Spain. According to the European microbiological criteria (92/46/EEC, 93/43/EEC, and Commission Regulation No. 2073/2005), 31% and 35% of the total examined lots exceed the adopted limits of mesophilic aerobic counts and Enterobacteriaceae, respectively. This may be due to incorrect handling of milk and dairy products and inadequate cleaning and sanitization of milk container. Further, 2% of all the lots examined were positive for the presence of Escherich…

The organoid era permits the development of new applications to study glioblastoma

Simple Summary Glioblastoma is the most lethal primary adult brain tumor. The great number of mutations involved and the aggressiveness of glioblastoma render this type of cancer especially difficult to investigate. To address this problem, cerebral organoids have emerged as promising tools to investigate brain biology and to recapitulates the major steps involved in glioblastoma tumorigenesis. This review focuses on methods of cerebral organoid development, describes the protocols used for inducing glioblastoma, the approach used to derive glioblastoma organoids directly from patients’ biopsies and discusses their limitations and potential future direction. Abstract Glioblastoma (GB) is th…

A survey of mycotoxins in random street-vended snacks from Lagos, Nigeria, using QuEChERS-HPLC-MS/MS

Abstract A survey in African snacks was carried out in order to evaluate the intake of 23 mycotoxins. The African snack samples were purchased from street vendors within Lagos metropolis (Nigeria) and evaluated for the presence of 23 mycotoxins using a modified QuEChERS procedure coupled with liquid chromatography-triple quadrupole linear ion trap mass spectrometer. The snacks included akara, baked coconut, coconut candy, donkwa, groundnut cake (kulikuli), lafun, milk curd (wara), fresh and dried tiger-nuts, and yam flour. Only three mycotoxins were detected in 23.8% of the studied snacks, and at concentrations ranging from 6 to 54 μg kg−1. The concentrations of aflatoxin B1 (AFB1) and AFB2…

Natural co-occurrence of mycotoxins in wheat grains from Italy and Syria

This article describes the application of an analytical method for the detection of 25 mycotoxins in wheat grain based on simultaneous extraction using matrix solid-phase dispersion (MSPD) followed by liquid chromatography coupled to tandem mass spectrometry, a hybrid triple quadrupole-linear ion trap mass spectrometer (QTrapÒ). Information Dependent Acquisition (IDA), an extra confirmation tool for samples that contain the target mycotoxins, was used. The analysis of 40 Syrian and 46 Italian wheat grain sam- ples interestingly showed that Syrian samples were mainly contaminated with ochratoxin A and aflatox- ins, whereas Italian samples with deoxynivalenol and 15-acetyldeoxynivalenol. Emer…

Mass spectrometry applications

The history of proteomics dates back to the discovery of two-dimensional gels in the 1970s, which provided the first feasible way of displaying hundreds or thousands of proteins on a single gel. Despite mass spectrometry being restricted for a long time to small and thermostable compounds, the development in the late 1980s of two techniques for the routine and general formation of molecular ions of intact biomolecules changed this situation and mass spectrometry has become an indispensable tool for proteomics research. The aim of this chapter is to review the major types of MS instruments used in proteomics analysis and to discuss strategies for the analysis of whole proteins and peptides o…

One-year monitoring of aflatoxins and ochratoxin A in tiger-nuts and their beverages.

A sensitive and selective liquid chromatography-triple quadrupole-tandem mass spectrometry (LC-ESI-MS-MS) method was developed for the routine analysis of aflatoxins (AFB(1), AFB(2), AFG(1) and AFG(2)) and ochratoxin A (OTA) in tiger nuts and tiger-nut beverage (horchata). A matrix solid phase dispersion was adapted to eliminate lipidic interferences. The solid support was C(18), while the elution solvent was acetonitrile. Mean recoveries obtained at two fortification levels were 72-83% and 71-81% for horchata and tiger nut respectively with relative standard deviations (RSDs) <13% and 15% respectively. The LC-MS-MS method allowed quantification and identification at low levels in two matri…

Evaluation of matrix solid-phase dispersion (MSPD) extraction for multi-mycotoxin determination in different flours using LC-MS/MS

An existing matrix solid-phase dispersion (MSPD) method for aflatoxins (AFs) and ochratoxin A (OTA) extraction was extended by further 14 mycotoxins. After it careful optimization, this method was applied to determine the occurrence of these mycotoxins on commercial flour samples (with different cereals composition) collected from local markets. In a total of 49 samples investigated, 9 mycotoxins were identified. Nivalenol (NIV) and Beauvericin (BEA) were the mycotoxins found most frequently. The samples that presented major contamination were wheat flours and bakery preparations. Despite of the great number of positives finding, only one wheat flour sample exceeded the maximum limits (ML) …

Rapid mycotoxin analysis in human urine: A pilot study

A simple and rapid method effective for quantitative determination of deoxynivalenol (DON), T-2 toxin (T-2), HT-2 toxin (HT-2), zearalenone (ZEN), ochratoxin A (OTA), aflatoxins (AFs) B(1), B(2), G(1) and G(2) and fumonisins FB(1) and FB(2) in urine was developed. The urine was diluted with phosphate buffer solution (PBS) and thoroughly mixed. For clean-up and extraction, the mixture was loaded on a MYCO 6in1 IAC. Hybrid triple quadrupole-linear ion trap mass spectrometer (QTrap) was used for the detection. Extra tools for confirmation of selected mycotoxins in positive samples, Information Dependent Acquisition (IDA) experiments, were also developed. The use of immunoaffinity columns follo…

Prevalence of bacteria and absence of anisakid parasites in raw and prepared fish and seafood dishes in Spanish restaurants

This study evaluated the presence of bacteria and anisakid parasites in 45 samples of raw anchovies in vinegar, a dish widely eaten in Spain, and in 227 samples of cooked fish and cephalopods served in Spanish food service establishments. Our analysis showed that, according to European and Spanish regulation, 14 to 30% of the prepared fish and cephalopod dishes exceeded the maximum allowable level for mesophilic aerobic counts, and 10 to 40% of these samples exceeded the allowable levels for Enterobacteriaceae. None of the studied samples showed evidence of anisakid parasites, Escherichia coli, Staphylococcus aureus, Salmonella, or Listeria monocyto genes. These results indicate that applic…

Analysis of mycotoxins in barley using ultra high liquid chromatography high resolution mass spectrometry: comparison of efficiency and efficacy of different extraction procedures.

The effectiveness of four extraction methods (modified QuEChERS, matrix solid-phase dispersion (MSPD), solid-liquid extraction (SLE) and solid-phase extraction (SPE) clean-up) were evaluated for simultaneous determination of 32 mycotoxins produced by the genus Fusarium, Claviceps, Aspergillus, Penicillium and Alternaria in barley by ultra high pressure liquid chromatography coupled to ultra-high resolution mass spectrometry (UHPLC-Orbitrap(®) MS). The efficiency and efficacy of extraction methods were evaluated and compared in number of extracted mycotoxins and obtained recoveries. From the one point of view, QuEChERS procedure was fast and easy, as well as it was able to successfully extra…

Risk-benefit in food safety and nutrition - outcome of the 2019 Parma Summer School

Risk-benefit assessment is the comparison of the risk of a situation to its related benefits, i.e. a comparison of scenarios estimating the overall health impact. The risk–benefit analysis paradigm mirrors the classical risk analysis one: risk–benefit assessment goes hand-in-hand with risk–benefit management and risk–benefit communication. The various health effects associated with food consumption, together with the increasing demand for advice on healthy and safe diets, have led to the development of different research disciplines in food safety and nutrition. In this sense, there is a clear need for a holistic approach, including and comparing all of the relevant health risks and benefit…

Untargeted metabolomics of fresh and heat treatment Tiger nut ( Cyperus esculentus L. ) milks reveals further insight into food quality and nutrition

Tiger nut (Cyperus esculentus L.) is a crop traditionally grown in Valencia Region (Spain) and other temperate and tropical regions in the world, where its tubers are commonly consumed as tiger nut milk (horchata). Because of their nutritive potential and original taste, these products are beginning to spread internationally and, as consequence, analytical procedures to assess nutritional profiles, quality control issues are acquiring increasing relevance. The main objective of this study was to use an advance analytical method and chemometrics tools to determine if the ultra-high temperature (UHT) treatment necessary to extend the shelf life of tiger nut milk would affect the profile of nu…

Study of mycotoxin calibration approaches on the example of trichothecenes analysis from flour

The aim of this study was to evaluate the applicability of different calibration approaches in trichothecenes analysis from wheat flour. Concretely eight trichothecenes (five type B trichothecenes and three type A trichothecenes) were analyzed by matrix solid-phase dispersion (MSPD) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In the first set of experiments the presence of matrix effects was evaluated; values ranged between 59% and 79%. In the second set of experiments, solutions to compensate these signal suppressions were examined. Different calibration methods showed to tackle matrix effects obtaining values between 69% and 85% for external matrix matched calibration a…

Optimization of Matrix Solid-Phase Dispersion method for simultaneous extraction of aflatoxins and OTA in cereals and its application to commercial samples

Abstract A method based on Matrix Solid-Phase Dispersion (MSPD) has been developed for the determination of 5 mycotoxins (ochratoxin A and aflatoxins B and G) in different cereals. Several dispersants, eluents and ratios were tested during the optimization of the process in order to obtain the best results. Finally, samples were blended with C 18 and the mycotoxins were extracted with acetonitrile. Regarding to matrix effects, the results clearly demonstrated the necessity to use a matrix-matched calibration to validate the method. Analyses were performed by liquid chromatography–triple quadrupole-tandem mass spectrometry (LC–QqQ-MS/MS). The recoveries of the extraction process ranged from …

Evaluation of mycotoxins and their metabolites in human breast milk using liquid chromatography coupled to high resolution mass spectrometry

Humans can be exposed to mycotoxins through the food chain. Mycotoxins are mainly found as contaminants in food and could be subsequently excreted via biological fluids such as urine or human breast milk in native or metabolised form. Since breast milk is usually supposed as the only food for new-borns, the occurrence of mycotoxins in thirty-five human milk samples was evaluated by a newly developed method based on QuEChERS extraction and UHPLC-HRMS detection. The method described here allows the detection of target mycotoxins in order to determine the quality of this initial feeding. The method has been fully validated, with recoveries ranging from 64% to 93% and relative standard deviatio…

Mass spectrometry strategies for mycotoxins analysis in European beers

In this work, an existent solid-phase extraction (SPE) procedure was used to study the occurrence of mycotoxins in different European beers. HPLC-QqQ-MS/MS and ultra high resolution mass analyser have been optimized for the analysis of 18 mycotoxins: the methods were validated according to the EU Commission Decision 2002/657/EC guidelines. In this sense, matrix-matched calibration was performed for each type of beer, obtaining an effective quantification. The recoveries ranged from 63 to 91% and repeatability and reproducibility expressed as relative standard deviation (RSD%) were lower than 17%. On one hand, HPLC-LTQ-Orbitrap ® was used for unambiguous identification of target mycotoxins, …