21 results on '"Berthiller F"'
Search Results
2. Characterization of (13C24) T-2 toxin and its use as an internal standard for the quantification of T-2 toxin in cereals with HPLC–MS/MS
- Author
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Häubl, G., Berthiller, F., Hametner, C., Rechthaler, J., Jaunecker, G., Freudenschuss, M., Krska, R., and Schuhmacher, R.
- Published
- 2007
- Full Text
- View/download PDF
3. Suitability of a fully 13C isotope labeled internal standard for the determination of the mycotoxin deoxynivalenol by LC-MS/MS without clean up
- Author
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Häubl, G., Berthiller, F., Krska, R., and Schuhmacher, R.
- Published
- 2006
- Full Text
- View/download PDF
4. Processing and purity assessment of standards for the analysis of type-B trichothecene mycotoxins
- Author
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Krska, R., Schothorst, R. C., van Egmond, H. P., Josephs, R. D., Lepschy, J., Pettersson, H., Chan, D., Berthiller, F., Schuhmacher, R., Kandler, W., Parich, A., and Welzig, E.
- Published
- 2005
- Full Text
- View/download PDF
5. Suitability of a fully 13C isotope labeled internal standard for the determination of the mycotoxin deoxynivalenol by LC-MS/MS without clean up
- Author
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Häubl, G., primary, Berthiller, F., additional, Krska, R., additional, and Schuhmacher, R., additional
- Published
- 2005
- Full Text
- View/download PDF
6. Comparison of LC-MS-based methods for the determination of carboxylic acids in animal matrices.
- Author
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Schwartz-Zimmermann HE, Hündler M, Reiterer N, Ricci S, Rivera-Chacon R, Castillo-Lopez E, Zebeli Q, and Berthiller F
- Subjects
- Humans, Female, Animals, Cattle, Chromatography, Liquid methods, Liquid Chromatography-Mass Spectrometry, Chromatography, High Pressure Liquid methods, Aniline Compounds, Carboxylic Acids chemistry, Tandem Mass Spectrometry methods
- Abstract
Carboxylic acids (CAs) are key players in human and animal metabolism. As they are hardly retained under reversed-phase liquid chromatography (RP-LC) conditions in their native form, derivatization is an option to make them accessible to RP-LC and simultaneously increase their response for mass spectrometric detection. In this work, two RP-LC tandem mass spectrometry-based methods using aniline or 3-nitrophenylhydrazine (3-NPH) as derivatization agents were compared with respect to several factors including completeness of derivatization, apparent recoveries (R
A s) in both cow feces and ruminal fluid, and concentrations obtained in feces and ruminal fluid of cows. Anion exchange chromatography coupled to high-resolution mass spectrometry (AIC-HR-MS) served as reference method. Derivatization efficiencies were close to 100% for 3-NPH derivatization but variable (20-100%) and different in solvent solutions and matrix extracts for aniline derivatization. Likewise, average RA s of13 C-labeled short-chain fatty acids as internal standards were around 100% for 3-NPH derivatization but only 45% for aniline derivatization. Quantification of CAs in feces and ruminal fluid of cows initially fed a forage-only diet and then transitioned to a 65% high-grain diet which yielded similar concentrations for 3-NPH derivatization and AIC-HR-MS, but concentrations determined by aniline derivatization were on average five times lower. For these reasons, derivatization with aniline is not recommended for the quantitative analysis of CAs in animal samples., (© 2024. The Author(s).)- Published
- 2024
- Full Text
- View/download PDF
7. Novel analytical methods to study the fate of mycotoxins during thermal food processing.
- Author
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Stadler D, Berthiller F, Suman M, Schuhmacher R, and Krska R
- Subjects
- Food Contamination analysis, Food Handling methods, Mycotoxins analysis
- Abstract
Food processing can lead to a reduction of contaminants, such as mycotoxins. However, for food processing operations where thermal energy is employed, it is often not clear whether a reduction of mycotoxins also results in a mitigation of the toxicological impact. This is often due to the reason that the formed degradation products are not characterized and data on their toxicity is scarce. From the perspective of an analytical chemist, the elucidation of the fate of a contaminant in a complex food matrix is extremely challenging. An overview of the analytical approaches is given here, and the application and limitations are exemplified based on cases that can be found in recent literature. As most studies rely on targeted analysis, it is not clear whether the predetermined set of compounds differs from the degradation products that are actually formed during food processing. Although untargeted analysis allows for the elucidation of the complete spectrum of degradation products, only one such study is available so far. Further pitfalls include insufficient precision, natural contamination with masked forms of mycotoxins and interferences that are caused by the food matrix. One topic that is of paramount importance for both targeted and untargeted approaches is the availability of reference standards to identity and quantity the formed degradation products. Our vision is that more studies need to be published that characterize the formed degradation products, collect data on their toxicity and thereby complete the knowledge about the mycotoxin mitigating effect during food processing.
- Published
- 2020
- Full Text
- View/download PDF
8. The contribution of lot-to-lot variation to the measurement uncertainty of an LC-MS-based multi-mycotoxin assay.
- Author
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Stadler D, Sulyok M, Schuhmacher R, Berthiller F, and Krska R
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- Calibration, Ficus chemistry, Reference Standards, Reproducibility of Results, Zea mays chemistry, Chromatography, Liquid methods, Mycotoxins analysis, Spectrometry, Mass, Electrospray Ionization methods, Tandem Mass Spectrometry methods, Uncertainty
- Abstract
Multi-mycotoxin determination by LC-MS is commonly based on external solvent-based or matrix-matched calibration and, if necessary, the correction for the method bias. In everyday practice, the method bias (expressed as apparent recovery RA), which may be caused by losses during the recovery process and/or signal/suppression enhancement, is evaluated by replicate analysis of a single spiked lot of a matrix. However, RA may vary for different lots of the same matrix, i.e., lot-to-lot variation, which can result in a higher relative expanded measurement uncertainty (U
r ). We applied a straightforward procedure for the calculation of Ur from the within-laboratory reproducibility, which is also called intermediate precision, and the uncertainty of RA (ur,RA ). To estimate the contribution of the lot-to-lot variation to Ur , the measurement results of one replicate of seven different lots of figs and maize and seven replicates of a single lot of these matrices, respectively, were used to calculate Ur . The lot-to-lot variation was contributing to ur,RA and thus to Ur for the majority of the 66 evaluated analytes in both figs and maize. The major contributions of the lot-to-lot variation to ur,RA were differences in analyte recovery in figs and relative matrix effects in maize. Ur was estimated from long-term participation in proficiency test schemes with 58%. Provided proper validation, a fit-for-purpose Ur of 50% was proposed for measurement results obtained by an LC-MS-based multi-mycotoxin assay, independent of the concentration of the analytes.- Published
- 2018
- Full Text
- View/download PDF
9. Tracing the metabolism of HT-2 toxin and T-2 toxin in barley by isotope-assisted untargeted screening and quantitative LC-HRMS analysis.
- Author
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Meng-Reiterer J, Varga E, Nathanail AV, Bueschl C, Rechthaler J, McCormick SP, Michlmayr H, Malachová A, Fruhmann P, Adam G, Berthiller F, Lemmens M, and Schuhmacher R
- Subjects
- Chromatography, Liquid methods, Tandem Mass Spectrometry methods, Fusarium metabolism, Hordeum metabolism, Hordeum microbiology, T-2 Toxin analogs & derivatives, T-2 Toxin metabolism
- Abstract
An extensive study of the metabolism of the type A trichothecene mycotoxins HT-2 toxin and T-2 toxin in barley using liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) is reported. A recently developed untargeted approach based on stable isotopic labelling, LC-Orbitrap-MS analysis with fast polarity switching and data processing by MetExtract software was combined with targeted LC-Q-TOF-MS(/MS) analysis for metabolite structure elucidation and quantification. In total, 9 HT-2 toxin and 13 T-2 toxin metabolites plus tentative isomers were detected, which were successfully annotated by calculation of elemental formulas and further LC-HRMS/MS measurements as well as partly identified with authentic standards. As a result, glucosylated forms of the toxins, malonylglucosides, and acetyl and feruloyl conjugates were elucidated. Additionally, time courses of metabolite formation and mass balances were established. For absolute quantification of those compounds for which standards were available, the method was validated by determining apparent recovery, signal suppression, or enhancement and extraction recovery. Most importantly, T-2 toxin was rapidly metabolised to HT-2 toxin and for both parent toxins HT-2 toxin-3-O-β-glucoside was identified (confirmed by authentic standard) as the main metabolite, which reached its maximum already 1 day after toxin treatment. Graphical Abstract Isotope-assisted untargeted screening of HT-2 toxin and T-2 toxin metabolites in barley.
- Published
- 2015
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10. Critical evaluation of indirect methods for the determination of deoxynivalenol and its conjugated forms in cereals.
- Author
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Malachová A, Štočková L, Wakker A, Varga E, Krska R, Michlmayr H, Adam G, and Berthiller F
- Subjects
- Chromatography, Liquid methods, Edible Grain microbiology, Hordeum chemistry, Hordeum microbiology, Hydrolysis, Tandem Mass Spectrometry methods, Triticum chemistry, Triticum microbiology, Zea mays chemistry, Zea mays microbiology, Edible Grain chemistry, Glucosides analysis, Mycotoxins analysis, Trichothecenes analysis
- Abstract
A critical assessment of three previously published indirect methods based on acidic hydrolysis using superacids for the determination of "free" and "total" deoxynivalenol (DON) was carried out. The modified mycotoxins DON-3-glucoside (D3G), 3-acetyl-DON (3ADON), and 15-acetyl-DON (15ADON) were chosen as model analytes. The initial experiments focused on the stability/degradation of DON under hydrolytic conditions and the ability to release DON from the modified forms. Acidic conditions that were capable of cleaving D3G, 3ADON, and 15ADON to DON were not found, raising doubts over the efficacy of previously published indirect methods for total DON determination. Validation of these indirect methods for wheat, maize, and barley using UHPLC-MS/MS was performed in order to test the accuracy of the generated results. Validation data for DON, D3G, 3ADON, and 15ADON in nonhydrolyzed and hydrolyzed matrices were obtained. Under the tested conditions, DON was not released from D3G, 3ADON, or 15ADON after hydrolysis and thus none of the published methods were able to cleave the modified forms of DON. In addition to acids, alkaline hydrolysis with KOH for an extended time and at elevated temperatures was also tested. 3ADON and 15ADON were cleaved under the alkaline pH caused by the addition of KOH or aqueous K2CO3 to "neutralize" the acidic sample extracts in the published studies. The published additional DON increase after hydrolysis may have been caused by huge differences in matrix effects and the recovery of DON in nonhydrolyzed and hydrolyzed matrices as well as by the alkaline cleavage of 3ADON or 15ADON after the neutralization of hydrolyzed extracts.
- Published
- 2015
- Full Text
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11. Simultaneous determination of major type A and B trichothecenes, zearalenone and certain modified metabolites in Finnish cereal grains with a novel liquid chromatography-tandem mass spectrometric method.
- Author
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Nathanail AV, Syvähuoko J, Malachová A, Jestoi M, Varga E, Michlmayr H, Adam G, Sieviläinen E, Berthiller F, and Peltonen K
- Subjects
- Finland, Chromatography, Liquid methods, Edible Grain chemistry, Tandem Mass Spectrometry methods, Trichothecenes analysis, Zearalenone analysis
- Abstract
A reliable and sensitive liquid chromatography-tandem mass spectrometric method was developed for the simultaneous quantitative determination in cereals of the Fusarium mycotoxins HT-2 toxin, T-2 toxin, deoxynivalenol, nivalenol and zearalenone, as well as the modified metabolites 3-acetyl-deoxynivalenol, α-zearalenol, β-zearalenol, deoxynivalenol-3-glucoside, HT-2-3-glucoside, nivalenol-3-glucoside, zearalenone-14-glucoside, zearalenone-14-sulphate, zearalenone-16-glucoside, α-zearalenol-14-glucoside and β-zearalenol-14-glucoside. The 'dilute and shoot' approach was used for sample preparation after extraction with acetonitrile:water:acetic acid (79:20:1, v/v/v). Separation was carried out using reversed-phase liquid chromatography, and detection was performed using tandem mass spectrometry in the selected reaction monitoring mode. The method was in-house validated according to performance characteristics, established in Commission Regulation EC No 401/2006 and Commission Decision EC No 657/2002, prior to its application in a nationwide survey for the analysis of barley, oat and wheat samples (n = 95) harvested in Finland during 2013. Deoxynivalenol and its glucosylated form were the most abundant of the analytes, being detected in 93 and 81 % of the samples, respectively. Concentrations of deoxynivalenol were unusually high in 2013, especially in oats, with some cases exceeding the maximum legislative limits for unprocessed oats placed on the market for first-stage processing. All modified mycotoxins analysed were detected, and the natural occurrence of some of these compounds (e.g. zearalenone-16-glucoside and nivalenol-3-glucoside) in barley, oats and/or wheat was documented for the first time.
- Published
- 2015
- Full Text
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12. Deoxynivalenol (DON) sulfonates as major DON metabolites in rats: from identification to biomarker method development, validation and application.
- Author
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Schwartz-Zimmermann HE, Hametner C, Nagl V, Slavik V, Moll WD, and Berthiller F
- Subjects
- Animals, Chromatography, Liquid methods, Feces chemistry, Feces microbiology, Limit of Detection, Male, Mycotoxins metabolism, Mycotoxins urine, Rats, Rats, Sprague-Dawley, Sulfonic Acids metabolism, Sulfonic Acids urine, Trichothecenes metabolism, Trichothecenes urine, Mycotoxins analysis, Sulfonic Acids analysis, Tandem Mass Spectrometry methods, Trichothecenes analysis
- Abstract
Deoxynivalenol (DON) is a trichothecene mycotoxin regularly occurring in cereals. Rats are often used to study toxicokinetics of DON and related compounds, yet only about 30 % of the administered dose is typically recovered. Recently, it was reported that DON is partly metabolised to previously undetected DON- and deepoxy-DON (DOM) sulfonate in rats and tentative structures were proposed. The present work describes the production and characterisation of DON-, DOM- and DON-3-glucoside (D3G) sulfonates of three different series; the development and validation of liquid chromatography tandem mass spectrometry (LC-MS/MS)-based methods for determination of DON, DOM, D3G and their sulfonates in rat faeces and urine; and application of the methods to samples from a DON and D3G feeding trial with rats. In addition to previously produced DON sulfonates (DONS) 1, 2 and 3, D3G sulfonates 1, 2 and 3; and DOM sulfonates (DOMS) 2 and 3 were synthesised, purified and characterised. The developed methods showed apparent recoveries of all investigated compounds between 68 and 151 % in faeces and between 48 and 113 % in urine. The recovery of DON, D3G and their metabolites from faeces and urine of rats (n = 6) administered in a single dose of 2.0 mg/kg b.w. DON or the equimolar amount of D3G was 75 ± 9 % for the DON group and 68 ± 8 % for the D3G group. DON-, DOM- and D3G sulfonates excreted in faeces accounted for 48 and 47 % of the total amount of administered DON and D3G. Urinary excretion of sulfonates was <1 %. In both treatment groups, DONS 2 was the major metabolite 0-24 h after treatment, whereas DOMS 2 was predominant thereafter. The developed methods can also be used for investigation of DON (conjugate) sulfonate formation in other animal species.
- Published
- 2014
- Full Text
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13. Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed.
- Author
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Peters J, Thomas D, Boers E, de Rijk T, Berthiller F, Haasnoot W, and Nielen MW
- Subjects
- Chromatography, Liquid, Color, Fumonisins chemistry, Microspheres, Ochratoxins chemistry, Reproducibility of Results, Tandem Mass Spectrometry, Zearalenone chemistry, Animal Feed analysis, Edible Grain chemistry, Flow Cytometry, Food Contamination analysis, Food Safety methods, Fumonisins analysis, Immunoassay, Ochratoxins analysis, Zearalenone analysis
- Abstract
A combined (triplex) immunoassay for the simultaneous detection of three mycotoxins in grains was developed with superparamagnetic colour-encoded microbeads, in combination with two bead-dedicated flow cytometers. Monoclonal antibodies were coupled to the beads, and the amounts of bound mycotoxins were inversely related to the amounts of bound fluorescent labelled mycotoxins (inhibition immunoassay format). The selected monoclonal antibodies were tested for their target mycotoxins and for cross-reactivity with relevant metabolites and masked mycotoxins. In the triplex format, low levels of cross-interactions between the assays occurred at irrelevant high levels only. All three assays were influenced by the sample matrix of cereal extracts to some extent, and matrix-matched calibrations are recommended for quantitative screening purposes. In a preliminary in-house validation, the triplex assay was found to be reproducible, sensitive and sufficiently accurate for the quantitative screening at ML level. The triplex assay was critically compared to liquid chromatography-tandem mass spectrometry using reference materials and fortified blank material. Results for the quantification of ochratoxin A and zearalenone were in good agreement. However, the fumonisin assay was, due to overestimation, only suitable for qualitative judgements. Both flow cytometer platforms (Luminex 100 and FLEXMAP 3D) performed similar with respect to sensitivity with the advantages of a higher sample throughput and response range of the FLEXMAP 3D and lower cost of the Luminex 100.
- Published
- 2013
- Full Text
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14. Development and validation of a (semi-)quantitative UHPLC-MS/MS method for the determination of 191 mycotoxins and other fungal metabolites in almonds, hazelnuts, peanuts and pistachios.
- Author
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Varga E, Glauner T, Berthiller F, Krska R, Schuhmacher R, and Sulyok M
- Subjects
- Arachis chemistry, Corylus chemistry, Food Contamination, Pistacia chemistry, Prunus chemistry, Reproducibility of Results, Chromatography, High Pressure Liquid methods, Food Analysis methods, Mycotoxins chemistry, Nuts chemistry, Tandem Mass Spectrometry methods
- Abstract
A multi-target method for the determination of 191 fungal metabolites in almonds, hazelnuts, peanuts and pistachios was developed. The method includes all mycotoxins regulated in the European Union and mycotoxins regularly found in food. After extraction with an acidified acetonitrile water mixture, the raw extract was diluted and injected directly into the UHPLC-MS/MS system. In two chromatographic runs, analysis was performed in positive and in negative ionisation mode. The method was in-house validated for the most important 65 analytes in these four commodities. Apparent recoveries between 80 and 120% were obtained for about half of the analyte-matrix combinations. Good repeatabilities (standard deviations < 10%) were achieved for the vast majority (83%) of all cases. Only in 6% of all combinations did the standard deviations exceed 15%. Matrix effects, arising during electrospray ionisation, significantly influenced the determination. For instance, signal suppression was observed for several early-eluting analytes and also signal enhancement up to 295% for physcion in peanuts was determined. Concerning extraction recovery, 94% of the analyte-matrix combinations showed values higher than 50%. Lower limits of quantification ranged between 0.04 μg kg(-1) for enniatin B3 in peanuts and 500 μg kg(-1) for HC toxin in hazelnuts. Additionally, the applicability of the developed method was demonstrated through the analysis of 53 naturally contaminated nut samples from Austria and Turkey. Overall, 40 toxins were quantified; the most frequently found mycotoxins were beauvericin (79%), enniatin B (62%) and macrosporin (57%). In the most contaminated hazelnut sample, 26 different fungal metabolites were detected.
- Published
- 2013
- Full Text
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15. Stable isotopic labelling-assisted untargeted metabolic profiling reveals novel conjugates of the mycotoxin deoxynivalenol in wheat.
- Author
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Kluger B, Bueschl C, Lemmens M, Berthiller F, Häubl G, Jaunecker G, Adam G, Krska R, and Schuhmacher R
- Subjects
- Chromatography, Liquid methods, Fusarium metabolism, Molecular Structure, Tandem Mass Spectrometry methods, Triticum chemistry, Isotope Labeling methods, Metabolomics methods, Trichothecenes chemistry, Trichothecenes metabolism, Triticum microbiology
- Abstract
An untargeted screening strategy for the detection of biotransformation products of xenobiotics using stable isotopic labelling (SIL) and liquid chromatography-high resolution mass spectrometry (LC-HRMS) is reported. The organism of interest is treated with a mixture of labelled and non-labelled precursor and samples are analysed by LC-HRMS. Raw data are processed with the recently developed MetExtract software for the automated extraction of corresponding peak pairs. The SIL-assisted approach is exemplified by the metabolisation of the Fusarium mycotoxin deoxynivalenol (DON) in planta. Flowering ears were inoculated with 100 μg of a 1 + 1 (v/v) mixture of non-labelled and fully labelled DON. Subsequent sample preparation, LC-HRMS measurements and data processing revealed a total of 57 corresponding peak pairs, which originated from ten metabolites. Besides the known DON and DON-3-glucoside, which were confirmed by measurement of authentic standards, eight further DON-biotransformation products were found by the untargeted screening approach. Based on a mass deviation of less than ±5 ppm and MS/MS measurements, one of these products was annotated as DON-glutathione (GSH) conjugate, which is described here for the first time for wheat. Our data further suggest that two DON-GSH-related metabolites, the processing products DON-S-cysteine and DON-S-cysteinyl-glycine and five unknown DON conjugates were formed in planta. Future MS/MS measurements shall reveal the molecular structures of the detected conjugates in more detail.
- Published
- 2013
- Full Text
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16. Stable isotope dilution assay for the accurate determination of mycotoxins in maize by UHPLC-MS/MS.
- Author
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Varga E, Glauner T, Köppen R, Mayer K, Sulyok M, Schuhmacher R, Krska R, and Berthiller F
- Subjects
- Tandem Mass Spectrometry methods, Carbon Isotopes analysis, Chromatography, High Pressure Liquid methods, Food Contamination analysis, Indicator Dilution Techniques, Mycotoxins analysis, Spectrometry, Mass, Electrospray Ionization methods, Zea mays chemistry
- Abstract
A fast, easy-to-handle and cost-effective analytical method for 11 mycotoxins currently regulated in maize and other cereal-based food products in Europe was developed and validated for maize. The method is based on two extraction steps using different acidified acetonitrile–water mixtures. Separation is achieved using ultrahigh-performance liquid chromatography (UHPLC) by a linear water–methanol gradient. After electrospray ionisation, tandem mass spectrometric detection is performed in dynamic multiple reaction monitoring mode. Since accurate mass spectrometric quantification is hampered by matrix effects, uniformly [(13)C]-labelled mycotoxins for each of the 11 compounds were added to the sample extracts prior to UHPLC-MS/MS analysis. Method performance parameters were obtained by spiking blank maize samples with mycotoxins before as well as after extraction on six levels in triplicates. The twofold extraction led to total recoveries of the extraction steps between 97% and 111% for all target analytes, including fumonisins. The [(13)C]-labelled internal standards efficiently compensated all matrix effects in electrospray ionisation, leading to apparent recoveries between 88% and 105% with reasonable additional costs. The relative standard deviations of the whole method were between 4% and 11% for all analytes. The trueness of the method was verified by the measurement of several maize test materials with well-characterized concentrations. In conclusion, the developed method is capable of determining all regulated mycotoxins in maize and presuming similar matrix effects and extraction recovery also in other cereal-based foods.
- Published
- 2012
- Full Text
- View/download PDF
17. Direct quantification of deoxynivalenol glucuronide in human urine as biomarker of exposure to the Fusarium mycotoxin deoxynivalenol.
- Author
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Warth B, Sulyok M, Berthiller F, Schuhmacher R, Fruhmann P, Hametner C, Adam G, Fröhlich J, and Krska R
- Subjects
- Chromatography, Liquid economics, Chromatography, Liquid methods, Humans, Limit of Detection, Tandem Mass Spectrometry economics, Fusarium metabolism, Glucuronides urine, Mycotoxins urine, Tandem Mass Spectrometry methods, Trichothecenes urine, Urine microbiology
- Abstract
The direct quantification of deoxynivalenol glucuronide (DON-GlcA) by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and its application as a biomarker of exposure to the Fusarium mycotoxin deoxynivalenol (DON) is reported. Usually, DON exposure is estimated from dietary average intakes or by measurement of the native toxin in urine after enzymatic hydrolysis with β-glucuronidase. These methods are time-consuming, expensive, and fail to determine the ratio of DON to DON-GlcA in a simple one-step procedure. One of the main reasons for the use of indirect methods is the unavailability of DON-GlcA standards. Consequently, DON-3-O-glucuronide (D3GlcA) was synthesized and used to develop a method allowing quantification of both DON and D3GlcA by a simple "dilute and shoot" approach without the need for any cleanup. Limit of detection and apparent recovery of D3GlcA was 3 μg l(-1) and 88%, respectively. The identity of D3GlcA in human urine was confirmed by comparison with LC-MS/MS measurements of the synthetically produced D3GlcA standard which was also used for external calibration. The applicability of the method was demonstrated through the analysis of urine samples obtained from a volunteer during regular and cereal-restricted diet, respectively. In regular-diet urine samples, D3GlcA was quantified in concentrations >30 μg l(-1) by this approach.
- Published
- 2011
- Full Text
- View/download PDF
18. Update on analytical methods for toxic pyrrolizidine alkaloids.
- Author
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Crews C, Berthiller F, and Krska R
- Subjects
- Chromatography, High Pressure Liquid trends, Mass Spectrometry trends, Molecular Structure, Plants chemistry, Chromatography, High Pressure Liquid methods, Food Analysis methods, Mass Spectrometry methods, Pyrrolizidine Alkaloids chemistry, Toxins, Biological chemistry
- Abstract
Methods for the determination of toxic pyrrolizidine alkaloids in plants and foods are described with emphasis on the important aspects of sample extraction and clean-up and the now preferred determination by liquid chromatography-mass spectrometry. The efficiencies of different extraction solvents and methods are described, as are the methods of reduction of N-oxides. Appropriate liquid chromatography-mass spectrometry conditions are tabulated. This concise review is intended to guide analysts towards adopting a more unified and reliable approach to the analysis of these important toxins.
- Published
- 2010
- Full Text
- View/download PDF
19. A reference-gene-based quantitative PCR method as a tool to determine Fusarium resistance in wheat.
- Author
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Brunner K, Kovalsky Paris MP, Paolino G, Bürstmayr H, Lemmens M, Berthiller F, Schuhmacher R, Krska R, and Mach RL
- Subjects
- Biomass, DNA, Fungal genetics, DNA, Plant genetics, Plant Diseases microbiology, Triticum microbiology, DNA, Fungal analysis, DNA, Plant analysis, Fusarium genetics, Plant Diseases genetics, Polymerase Chain Reaction methods, Triticum genetics
- Abstract
In recent years, plant breeders made great progress in breeding Fusarium-tolerant wheat lines. However, total resistance to this genus of plant pathogenic fungi has not yet been achieved as the resistance genes are located on several distinct genetic regions. Visual scoring of disease symptoms in combination with the analysis of mycotoxins is commonly applied to assess the tolerance of new lines. Both approaches are indirect methods and do not mandatorily determine the accumulated fungal biomass. Quantitative PCR is a useful tool to assess fungal biomass based on the abundance of organism-specific DNA. The aim of this study was the development of a quantitative PCR assay for trichothecene-producing Fusarium species and to adapt this method for resistance assessment of wheat lines artificially infected with Fusarium graminearum and Fusarium culmorum. Several DNA-extraction methods for wheat samples were evaluated and optimized for downstream real-time PCR analysis and furthermore, a new reference-gene-based approach for more accurate quantification of Fusarium biomass in cereals is presented. The co-determination of a plant gene was used to compensate for unequal DNA-extraction efficiencies.
- Published
- 2009
- Full Text
- View/download PDF
20. Formation, determination and significance of masked and other conjugated mycotoxins.
- Author
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Berthiller F, Schuhmacher R, Adam G, and Krska R
- Subjects
- Animals, Chromatography, Liquid methods, Enzyme-Linked Immunosorbent Assay methods, Humans, Mass Screening methods, Mycotoxins chemistry, Animal Feed analysis, Food Contamination analysis, Mycotoxins analysis
- Abstract
Mycotoxins are secondary metabolites of fungi poisonous for humans or animals which can be found on a great variety of food and feed commodities. Food is not necessarily safe just because the presence of well-known mycotoxins has been ruled out, as they might still be there in disguise. Mycotoxins may also occur in conjugated form, either soluble (masked mycotoxins) or incorporated into/associated with/attached to macromolecules (bound mycotoxins). These conjugated mycotoxins can emerge after metabolization by living plants, fungi and mammals or after food processing. Awareness of such altered forms of mycotoxins is increasing, but reliable analytical methods, measurement standards and occurrence and toxicity data are still lacking. In this paper currently known conjugated mycotoxins, their formation and determination are reviewed. For the latter, liquid chromatography-(tandem) mass spectrometry or ELISA methods are employed with or without conversion to the parent mycotoxins. Sample preparation to transform the bound forms into soluble forms can involve enzymatic or acidic/alkaline treatment. Especially mycotoxins which are in contact with living plants in the field are prone to be metabolized. This transformation process is not only important regarding food safety but also for the resistance of plants towards fungal-induced diseases, such as Fusarium head blight of wheat.
- Published
- 2009
- Full Text
- View/download PDF
21. Difficulties in fumonisin determination: the issue of hidden fumonisins.
- Author
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Dall'Asta C, Mangia M, Berthiller F, Molinelli A, Sulyok M, Schuhmacher R, Krska R, Galaverna G, Dossena A, and Marchelli R
- Subjects
- Chemistry Techniques, Analytical, Fumonisins chemistry, Molecular Structure, Fumonisins analysis, Zea mays chemistry
- Abstract
In this paper, the results obtained by five independent methods for the quantification of fumonisins B(1), B(2), and B(3) in raw maize are reported. Five naturally contaminated maize samples and a reference material were analyzed in three different laboratories. Although each method was validated and common calibrants were used, a poor agreement about fumonisin contamination levels was obtained. In order to investigate the interactions among analyte and matrix leading to this lack of consistency, the occurrence of fumonisin derivatives was checked. Significant amounts of hidden fumonisins were detected for all the considered samples. Furthermore, the application of an in vitro digestion protocol to raw maize allowed for a higher recovery of native fumonisins, suggesting that the interaction occurring among analytes and matrix macromolecules is associative rather than covalent. Depending on the analytical method as well as the maize sample, only 37-68% of the total fumonisin concentrations were found to be extractable from the samples. These results are particularly impressive and significant in the case of the certified reference material, underlying the actual difficulties in ascertaining the trueness of a method for fumonisin determination, opening thus an important issue for risk assessment.
- Published
- 2009
- Full Text
- View/download PDF
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