Levels of T-2 toxin and its metabolites, and the occurrence of Fusarium fungi in spring barley in the Czech Republic.

Mycotoxins have been widely studied by many research groups but further multidisciplinary research is needed to better understand and clarify many issues. This study describes the use of high-performance liquid chromatography coupled with ion trap mass spectrometry (HPLC-MS) to measure T-2 toxin and its metabolites, such as HT-2 toxin, neosolaniol (NEO) and diacetoxyscirpenol (DAS), as well as masked glucosylated mycotoxins in Fusarium-infected Czech spring barley. In total, 152 spring barley samples from the 2018 harvest were analyzed by the ELISA screening method for the presence of T-2 toxin. The most contaminated samples (15), which exceeded the recommended maximum level set by the EU for the sum of T-2 and HT-2 toxin in unprocessed cereals (200 µg/kg), were analyzed by HPLC-MS/MS and microbiological testing. Isolated fungi were evaluated microscopically and identified by polymerase chain reaction (PCR) assays. The prevalence of Fusarium species in spring barley across the Czech Republic in 2018 showed a predominance of F. poae (12 barley samples) and F. tricinctum (9 barley samples). Other strains (F. sporotrichioides and F. langsethiae) were present at a lower frequency, in 1 and 2 samples, respectively. The average concentration of T-2 plus HT-2 toxin was 107.7 µg/kg, while NEO and DAS were found in a few samples at values close to their limit of quantification. HT-2 glucoside was identified in all samples.

Determination of mycotoxins and their dietary exposure assessment in pale lager beers using immunoaffinity columns and UPLC-MS/MS.

The use of contaminated raw materials can lead to the transfer of mycotoxins into the final product, including beer. This study describes the use of the commercially available immunoaffinity column 11+Myco MS-PREP® and UPLC-MS/MS for the determination of mycotoxins in pale lager-type beers brewed in Czech Republic and other European countries. The additional aim of the work was to develop, optimize and validate this analytical method. Validation parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), precision and accuracy were tested. The calibration curves were linear with correlation coefficients (R2 > 0.99) for all mycotoxins under investigation. The LOD ranged from 0.1 to 50 ng/L and LOQ from 0.4 to 167 ng/L. Recoveries of the selected analytes ranged from 72.2 to 101.1%, and the relative standard deviation under conditions repeatability (RSDr) did not exceed 16.3% for any mycotoxin. The validated procedure was successfully applied for the analysis of mycotoxins in a total of 89 beers from the retail network. The results were also processed using advanced chemometric techniques and compared with similar published studies. The toxicological impact was taken into account.

Determination of T-2 and HT-2 Toxins in Seed of Milk Thistle [Silybum marianum (L.) Gaertn.] Using Immunoaffinity Column by UPLC-MS/MS.

Milk thistle [Silybum marianum (L.) Gaertn.] achieved a significant increase in interest over the past few years from local and foreign pharmaceutical corporations. The silymarin complex of constituents extracted from milk thistle achenes provides compelling health benefits primarily thanks to antioxidant activities and hepatoprotective effects. However, consuming mycotoxin-contaminated plant material can cause immunosuppression and hepatotoxic problems. The aim of this study was to develop and validate a method for the determination of mycotoxin content in milk thistle. Fusarium toxins as T-2 and HT-2 toxins in grown milk thistle harvested from a breeding station in the Czech Republic during 2020-2021 were studied. The analysis of T-2 and HT-2 toxins was performed by UPLC-MS/MS after immunoaffinity columns EASI-EXTRACT® T-2 & HT-2 clean up. All analysed samples of milk thistle were contaminated with T-2 toxin and HT-2 toxin. The content of T-2 toxin in the samples from 2020 was in the range of 122.7-290.2 µg/kg and HT-2 toxin 157.0-319.0 µg/kg. In 2021, the content of T-2 toxin was in the range of 28.8-69.9 µg/kg and HT-2 toxin was 24.2-75.4 µg/kg. The results show that the climatic conditions of the year of harvesting have a highly statistically significant effect on the content of T-2 and HT-2 toxins in milk thistle.