A Dilute-and-Shoot UHPLC-MS/MS Isotope Dilution Method for Simultaneous Determination and Confirmation of Eleven Mycotoxins in Dried Distillers Grains with Solubles

2021 ◽  
Author(s):  
Cristina B Nochetto ◽  
Li Hui
Keyword(s):  
Ochratoxin A ◽  
High Performance ◽  
Animal Feed ◽  
Animal Health ◽  
Fumonisin B1 ◽  
Isotope Dilution Method ◽  
Dilution Method ◽  
Distillers Grains With Solubles ◽  
Distiller's Grains ◽  
Relative Standard

Abstract Background Natural contamination of mycotoxins in dried distiller’s grains with solubles (DDGS) as a mainstream animal feed ingredient poses risk to animal health. Objective A regulatory method was needed for the agency to simultaneously detect eleven mycotoxins of high regulatory priority in DDGS. Methods Ten grams of DDGS sample were extracted twice with acetonitrile/water under mildly acidic condition. Two aliquots from the combined crude extract were taken and processed separately: (1) diluted 400-fold with solvent for analysis of deoxynivalenol and fumonisins B1 and B2; (2) pH adjusted to 7.5, then diluted 15.7-fold for analysis of aflatoxins B1, B2, G1, G2, ochratoxin A, zearalenone, and T-2 and HT-2 toxins. Uniformly-labelled 13C-isotopologues of these mycotoxins were added as internal standards to the diluted extracts for quantitative analysis by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC–MS/MS). Results. The linear quantitation ranges (µg/kg) were: aflatoxin B1, B2, G1, and G2, 1.57 to 105; zearalenone, 16.3 to 1090; T-2 toxin, 3.14 to 208; HT-2 toxin, 48.2 to 3220; ochratoxin A, 0.47 to 31.4; deoxynivalenol, 240 to 16000; fumonisin B1 and B2, 320 to 21200. Accuracies for these analytes at each of three fortification levels range from 70.7% to 100%, with corresponding relative standard deviations between 1.4% to 10.5%. True recoveries were all higher than 83%. Conclusions This method was successfully validated to meet the agency’s performance guidelines for regulatory methods. Highlights This method is easy, quick and robust to simultaneously quantify and confirm presence of eleven regulated mycotoxins in DDGS.

scholarly journals Multiresidue Mycotoxin Analysis in Corn Grain by Column High-Performance Liquid Chromatography with Postcolumn Photochemical and Chemical Derivatization: Single-Laboratory Validation

2009 ◽  
Vol 92 (1) ◽  
pp. 15-25 ◽  
Author(s):  
Maria Ofitserova ◽  
Sareeta Nerkar ◽  
Michael Pickering ◽  
Laszlo Torma ◽  
Nancy Thiex
Keyword(s):  
Ochratoxin A ◽  
High Performance ◽  
Solid Phase ◽  
Fumonisin B1 ◽  
Feed Additives ◽  
Performance Criteria ◽  
Method Performance ◽  
Relative Standard ◽  
Agricultural Materials ◽  
Corn Grain

Abstract A multiresidue method was developed and validated for simultaneous analysis of 5 families of mycotoxins in corn grain. Deoxynivalenol (DON); aflatoxins B1, B2, G1, G2; ochratoxin A; zearalenone; and fumonisins FB1 and FB2 are extracted from corn grain samples with watermethanol, and extracts are cleaned up using immunoaffinity and solid-phase extraction columns. The column high-performance liquid chromatographic method uses postcolumn photochemical derivatization for detection of aflatoxins and derivatization with o-phthalaldehyde for detection of fumonisins. Mean recoveries and relative standard deviation values () over studied fortification levels for the chosen matrixes were: DON 89.9, 8.7; aflatoxin B1 85, 9.4; aflatoxin B2 82.4, 9.7; aflatoxin G1 74.8, 13.5; aflatoxin G2 79.2, 10.0; fumonisin B1 96.2, 8.0; fumonisin B2 84.5, 6.4; zearalenone 91.7, 11.5; and ochratoxin A 87.4, 15.8. The method performance criteria, including specificity, accuracy, repeatability, operational range, and detection limits, were found to be within specifications set by the Feed Additives and Contaminants Group of the AOAC Agricultural Materials Community.

scholarly journals Simultaneous Rapid Detection of Aflatoxin B1 and Ochratoxin A in Spices Using Lateral Flow Immuno-Chromatographic Assay

Foods ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2738
Author(s):  
Xue Zhao ◽  
Xindi Jin ◽  
Zhang Lin ◽  
Qi Guo ◽  
Bin Liu ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Aflatoxin B1 ◽  
High Performance ◽  
Simultaneous Detection ◽  
High Sensitivity ◽  
Strong Stability ◽  
Test Strip ◽  
Chromatography Analysis ◽  
Relative Standard ◽  
Strip Test

Spices are susceptible to contamination by aflatoxin B1 (AFB1) and ochratoxin A (OTA), which are both mycotoxins with high toxicity and carcinogenicity. In this study, we aimed to develop an immuno-chromatographic strip test for the simultaneous quantification of AFB1 and OTA in spices by spraying the coupled antigens AFB1–ovalbumin (AFB1–OVA) and OTA–ovalbumin (OTA–OVA) on a nitrocellulose membrane. The test strip had high sensitivity, good specificity, and strong stability. The detection limits of these two mycotoxins in Chinese prickly ash, pepper, chili, cinnamon, and aniseed were 5 μg/kg. The false positivity rate was 2%, and the false negativity rate was 0%. The maximum coefficient of variation was 4.28% between batches and 5.72% within batches. The average recovery rates of AFB1 and OTA in spices were 81.2–113.7% and 82.2–118.6%, respectively, and the relative standard deviation (RSD) was <10%. The actual sample detection was consistent with high performance liquid chromatography analysis results. Therefore, the immuno-chromatographic test strips developed in this study can be used for the on-site simultaneous detection of AFB1 and OTA in spices. This method would allow the relevant regulatory agencies to strengthen supervision in an effort to reduce the possible human health hazards of such contaminated spices.

scholarly journals Liquid Chromatographic Determination of Deoxynivalenol in Baby Food and Animal Feed: Interlaboratory Study

2006 ◽  
Vol 89 (4) ◽  
pp. 1012-1020 ◽  
Author(s):  
Joerg Stroka ◽  
Michelle Derbyshire ◽  
Carsten Mischke ◽  
Massimo Ambrosio ◽  
Katy Kroeger ◽  
...  
Keyword(s):  
Standard Deviation ◽  
Relative Standard Deviation ◽  
High Performance ◽  
Animal Feed ◽  
Chromatographic Determination ◽  
Baby Food ◽  
Interlaboratory Study ◽  
Relative Standard ◽  
Liquid Chromatographic

Abstract An interlaboratory study was conducted for the determination of deoxynivalenol in baby food and animal feed by high-performance liquid chromatography with UV detection. The study included 14 participants representing a cross section of industry, official food control, and research facilities. Mean recoveries reported ranged from 89% (at 120 g/kg) to 85% (at 240 g/kg) for baby food and from 100% (at 200 g/kg) to 93% (at 400 g/kg) for animal feed. On the basis of the results for spiked samples (blind duplicates at 2 levels), as well as those for naturally contaminated samples (blind duplicates at 3 levels), the relative standard deviation for repeatability (RSDr) in analyses of baby food ranged from 6.4 to 14.0% and in analyses of animal feed, from 6.1 to 16.5%. The relative standard deviation for reproducibility (RSDR) in analyses of baby food ranged from 9.4 to 19.5% and in analyses of animal feed, from 10.5 to 25.2%. The HorRat values ranged from 0.4 to 1.0 and from 0.7 to 1.3, for baby food and animal feed, respectively. The method showed acceptable performance for within-laboratory and between-laboratory precision for each matrix, as required by European legislation.

scholarly journals Multiresidue method for the simultaneous analysis of antibiotics and mycotoxins in feeds by ultra-high performance liquid chromatography coupled to tandem mass spectrometry

2021 ◽  
Author(s):  
Ü.İ. Konak ◽  
H.A. Yatmaz ◽  
Ş. Nilüfer ◽  
T. Erkaymaz ◽  
M. Certel
Keyword(s):  
High Performance ◽  
Safety Issue ◽  
Animal Health ◽  
Animal Origin ◽  
Limit Of Quantification ◽  
Multiresidue Method ◽  
Animal Feeds ◽  
Relative Standard ◽  
Feed Samples

AbstractResidues in animal feeds and foods of animal origin have been important safety issue concerning both human and animal health. A multiresidue method for determination of eight mycotoxins and ten antibiotics was developed and validated in animal feeds by using QuEChERS (quick, easy, cheap, effective, rugged, and safe) extraction followed by UHPLC-MS/MS. Optimisation of UHPLC-MS/MS parameters was performed to achieve good separation and resolution. The method was validated according to the European Commission Decision 2002/657/EC. Matrix matched calibration curves showed good r2 (≥0.995) values, and limit of quantification (LOQ) values varied between 1.2 and 5.2 μg kg−1. Average recoveries ranged from 60 to 102% with relative standard deviations of 2.2 and 15.6% for all type of feed samples except for tetracyclines, lincomycin, tylosin, ochratoxin A, and fumonisin (B1 and B2).

scholarly journals Fusarium Species and Mycotoxins Contaminating Veterinary Diets for Dogs and Cats

2019 ◽  
Vol 7 (1) ◽  
pp. 26 ◽  
Author(s):  
Natalia Witaszak ◽  
Łukasz Stępień ◽  
Jan Bocianowski ◽  
Agnieszka Waśkiewicz
Keyword(s):  
Animal Feed ◽  
Fusarium Species ◽  
Fusarium Verticillioides ◽  
Animal Health ◽  
Companion Animals ◽  
Fumonisin B1 ◽  
Additional Risk ◽  
Two Samples ◽  
The Mean ◽  
The Eu

Veterinary diets are intended for diseased animals and may contain cereal grains, mainly maize and/or wheat. These, in turn, are often infected with pathogens of the Fusarium genus, which are able to produce numerous harmful mycotoxins. Forty-two samples of veterinary diets for dogs and cats were analyzed for the presence of Fusarium species and mycotoxins. Species were identified using molecular methods and the ergosterol and mycotoxins (fumonisin B1, deoxynivalenol, nivalenol and zearalenone) were quantified using HPLC methods. Two Fusarium species were identified: Fusarium proliferatum and Fusarium verticillioides. The highest concentrations of fumonisin B1, deoxynivalenol, nivalenol and zearalenone were 74.83, 2318.05, 190.90, and 45.84 ng/g, respectively. Only 9.5% of the samples were free from Fusarium mycotoxins. The acceptable limits of mycotoxin content in animal feed, specified by the EU regulations, were not exceeded in any of the samples tested. The mean mycotoxin content in veterinary diets for cats was lower than for dogs. Thus, it is recommended that veterinary diets are examined, since the mycotoxin contamination pose additional risk to animal health. The knowledge on Fusarium occurrence in veterinary diets is scarce and as far as we are aware this is the first report concerning the occurrence of Fusarium spp. and their important secondary metabolites—mycotoxins—in different types of veterinary diets for companion animals in Poland.

scholarly journals Comprehensive Evaluation of the Efficiency of Yeast Cell Wall Extract to Adsorb Ochratoxin A and Mitigate Accumulation of the Toxin in Broiler Chickens

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 37 ◽  
Author(s):  
Suvi Vartiainen ◽  
Alexandros Yiannikouris ◽  
Juha Apajalahti ◽  
Colm A. Moran
Keyword(s):  
Cell Wall ◽  
Yeast Cell ◽  
Ochratoxin A ◽  
Broiler Chickens ◽  
Animal Feed ◽  
Animal Health ◽  
Yeast Cell Wall ◽  
In Vivo Model

Ochratoxin A (OTA) is a common mycotoxin contaminant in animal feed. When absorbed from the gastrointestinal tract, OTA has a propensity for pathological effects on animal health and deposition in animal tissues. In this study, the potential of yeast cell wall extracts (YCWE) to adsorb OTA was evaluated using an in vitro method in which consecutive animal digestion events were simulated. Low pH markedly increased OTA binding to YCWE, which was reversed with a pH increased to 6.5. Overall, in vitro analysis revealed that 30% of OTA was adsorbed to YCWE. Additional computational molecular modelling revealed that change in pH alters the OTA charge and modulates the interaction with the YCWE β-d-glucans. The effectiveness of YCWE was tested in a 14-day broiler chicken trial. Birds were subjected to five dietary treatments; with and without OTA, and OTA combined with YCWE at three dosages. At the end of the trial, liver OTA deposition was evaluated. Data showed a decrease of up to 30% in OTA deposits in the liver of broilers fed both OTA and YCWE. In the case of OTA, a tight correlation between the mitigation efficacy of YCWE between in vitro and in vivo model could be observed.

scholarly journals Occurrence of ochratoxin A in animal tissues and feeds in Poland in 2014–2016

2017 ◽  
Vol 61 (4) ◽  
pp. 483-487 ◽  
Author(s):  
Katarzyna Pietruszka ◽  
Marta Piątkowska ◽  
Piotr Jedziniak
Keyword(s):  
Food Safety ◽  
Ochratoxin A ◽  
Animal Feed ◽  
Animal Health ◽  
Toxic Metabolite ◽  
Animal Tissues ◽  
Limit Of Quantification ◽  
Feed Analysis ◽  
Feed Samples

AbstractIntroduction: Ochratoxin A (OTA) is a toxic metabolite mainly produced by Aspergillus spp. and Penicillum spp. fungi. Research on the contamination of cereals, complete feeds, and tissues with this mycotoxin has indicated that it can be a toxicological problem impacting animal health and food safety in temperate climes. OTA contamination mainly besets the global pig industry, necessitating the monitoring of feeds and animal tissues. The aim of the study was to present the results of the official monitoring of OTA in animal tissues and feeds in Poland in 2014–2016 and determine the possible correlation between the presence of OTA in different types of samples.Material and Methods: The presence of ochratoxin A was determined using accepted procedures based on liquid chromatography with fluorescence detection after immunoaffinity column clean-up. Determination of OTA was afforded in the range of 0.3 μg/kg to 300 μg/kg in complete feeds and from 0.2 μg/kg to 150 μg/kg in the kidneys, liver, and muscles.Results: Over the three year span, about 23.5% of the animal tissues samples were contaminated by ochratoxin A. In the 2014 survey, 10% of the sample tissues contained 5–10 μg/kg (only one sample above 10 μg/kg), and in 2015 and 2016, 24% of samples showed levels above the limit of quantification 0.2 μg/kg, while none of the samples exceeded the established provisional action level of 5 μg/kg for animal tissues. The animal feed analysis showed that 9% was contaminated with ochratoxin A above the limit of quantification of 0.3μg/kg. In 2% of feed samples the OTA concentration was greater than 50 μg/kg.Conclusion: The results confirm the appropriacy of OTA contamination monitoring and help to increase food safety.

A quantitative UHPLC-MS/MS method for citrinin and ochratoxin A detection in food, feed and red yeast rice food supplements

2016 ◽  
Vol 9 (3) ◽  
pp. 343-352 ◽  
Author(s):  
J.A.L. Kiebooms ◽  
B. Huybrechts ◽  
C. Thiry ◽  
E.K. Tangni ◽  
A. Callebaut
Keyword(s):  
Risk Assessment ◽  
Ochratoxin A ◽  
High Performance ◽  
Food Supplements ◽  
Red Yeast Rice ◽  
Red Yeast ◽  
Relative Standard ◽  
Future Data ◽  
Food And Feed ◽  
Feed Samples

Mycotoxins may cause deleterious effects (among others nephrogenic, hepatogenic, carcinogenic, teratogenic, neurogenic) in animals and humans, therefore they have been intensely studied and monitored over the years. For citrinin (CIT), a nephrotoxic mycotoxin, however, this has not yet been the case. According to the latest European Food Safety Authority report, a correct risk assessment of CIT was not possible due to the lack of occurrence data. Besides, traces of CIT or its metabolite, dehydrocitrinone are widely (in up to 90% of samples) present in human urine according to recent Belgian and German scientific reports, which might imply chronic exposure. Only recently, a European maximum limit has been set for CIT in cholesterol reducing food supplements including red yeast fermented rice (RYR). During production of RYR through fungal (among others Monascus purpureus) fermentation of rice other components, like CIT, as well as nephrotoxic ochratoxin A (OTA) may form. Consequently, the present work attempted develop to a robust and routinely applicable ultra-high performance liquid chromatographytandem mass spectrometry (UHPLC-MS/MS) method for the analysis of CIT and OTA in food, feed and in RYR food supplements. The method was successfully validated based on EU/657/2002 and EU/519/2014 in RYR food supplements and wheat flour, achieving respective limits of quantification (LOQ) for CIT of 0.4 μg/kg and 0.1 μg/kg and for OTA of 15 μg/kg and 0.4 μg/kg. The average between-day recoveries varied from 72 to 110% with relative standard deviations ≤16%. Single-day validation in rice, curry and apple matrices showed LOQs ranging from 0.3-1.0 μg/kg. Next, the occurrence of CIT/OTA was surveyed in 138 RYR, food and feed samples, proving the potential of this method for future data acquisition within a risk assessment framework specifically for CIT, while also gaining information about the (co-)occurrence of OTA in edible matrices.

scholarly journals Standardization of an analytical method to quantify ochratoxin A in green coffee beans by high performance liquid chromatography

2020 ◽  
Vol 9 (8) ◽  
pp. e39985070
Author(s):  
Wilder Douglas Santiago ◽  
Alexandre Rezende Teixeira ◽  
Juliana de Andrade Santiago ◽  
Ana Cláudia Alencar Lopes ◽  
Rafaela Magalhães Brandão ◽  
...  
Keyword(s):  
Food Safety ◽  
Ochratoxin A ◽  
High Performance ◽  
Animal Health ◽  
Green Coffee ◽  
Fungal Contamination ◽  
High Performance Liquid Chromatograph ◽  
Liquid Chromatograph ◽  
Green Coffee Beans ◽  
Coffee Beans

Nowadays, Brazil is the largest producer and exporter of coffee, also the second largest consumer of the beverage. The importance of ensuring food safety for consumers has influenced research to improve and monitor the final product quality. Coffee is a product that presents a high risk of fungal contamination, which can result in the presence of mycotoxins and poses risks to human and animal health. Therefore, this study aimed to standardize a chromatographic method to test and quantify ochratoxin A in 13 samples of green coffee beans. The green coffee beans were stored in sheds without temperature or humidity control. Samples were ground, and the analyte was extracted by a 3% methanol:sodium bicarbonate (1:2 v/v) solution. Ochratoxin A was quantified in a high performance liquid chromatograph. The method was validated by testing the selectivity, linearity, accuracy, limits of detection and quantification. The method presented robustness to the tested parameters and among the analyzed samples. Ochratoxin A was detected above the limit established by the legislation (75.19 µg kg-1) only in one sample. Overall, the storage of green coffee beans in these sheds was adequate, since 12 samples had a low content of ochratoxin A and they were within the limit established by legislation. Therefore, food safety was guaranteed without any severe mycotoxin contamination.

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