Immunoaffinity columns Dotaz Zobrazit nápovědu
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.
Blood sausages consisting of groats, pork, porcine offal, fat, blood, and spices are very popular in the Czech Republic. All these ingredients are potential sources of dietary exposure to ochratoxin A (OTA). OTA has a strong affinity to serum proteins in porcine blood. Thus, the contamination of blood sausages with OTA can be expected. This study aims to evaluate OTA in 200 samples of porcine blood sausages purchased at the Czech market during 2020-2021. The analytical method high-performance liquid chromatography coupled with fluorescence detection with pre-treatment using immunoaffinity columns was employed to determine OTA. The limit of detection was 0.03 ng/g and the limit of quantification 0.10 ng/g. Recovery was 71.6 %. All samples were positive at contents ranging from 0.15 to 5.68 ng/g with a mean of 1.47 ng/g, and a median of 1.26 ng/g. A total of 66% of these samples contained OTA content exceeding the maximum limit of 1 ng/g set in Italy. This study demonstrates that the Czech population is exposed to OTA from blood sausages. The proposed preliminary action limit for OTA in blood sausages should be set at 1 ng/g. No regulatory limits for OTA in blood sausages have been established yet in the European Union legislation. To protect human health, further monitoring of OTA in these products is necessary.
- MeSH
- kontaminace potravin analýza MeSH
- masné výrobky * analýza MeSH
- ochratoxiny * analýza MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Specific allergen immunotherapy is frequently associated with adverse reactions. Several strategies are being developed to reduce the allergenicity while maintaining the therapeutic benefits. Peptide immunotherapy is one such approach. Methods for the simple and rapid identification of immunogenic epitopes of allergens (i.e. allergenic epitopes) are ongoing and could potentially lead to peptide-based vaccines. An epitope extraction technique, based on biofunctionalized magnetic microspheres self-organized under a magnetic field in a channel of a simple microfluidic device fabricated from polydimethylsiloxane, was applied in the isolation and identification of prospective allergenic epitopes. Similarly to chromatographic column separations, the easily replaceable plug of self-organized beads in the channel benefits especially from an even larger surface-to-volume ratio and an enhanced interaction of the surfaces with passing samples. Ovalbumin, the major protein of egg white and a typical representative of food allergens, was selected as the model molecule. Highly resistant ovalbumin was at first efficiently digested by a magnetic proteolytic reactor with trypsin treated with l-1-tosylamido-2-phenylethyl chloromethyl ketone and the second step, i.e. capture of allergenic epitopes from the mixture of peptides, was performed by a magnetic immunoaffinity carrier with orientedly immobilized rabbit anti-ovalbumin IgG molecules. Captured peptides were released with 0.05% trifluoroacetic acid. The elution fractions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The peptide fragment of ovalbumin HIATNAVLFFGR (m/z: 1345.75, position: 371-382) was identified as a relevant allergenic epitope in this way. Such a microfluidic magnetic force-based epitope extraction technique applied in the epitope mapping of ovalbumin has the potential to be a significant step towards developing safe and cost-effective epitope-based vaccines.
- MeSH
- alergeny chemie imunologie MeSH
- epitopy analýza MeSH
- financování organizované MeSH
- hmotnostní spektrometrie MeSH
- imunomagnetická separace metody MeSH
- mapování epitopu metody MeSH
- mikrofluidní analytické techniky metody MeSH
- mikrosféry MeSH
- ovalbumin chemie imunologie MeSH
- potravinová alergie MeSH
- vakcíny MeSH
