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.

• Klíčová slova
• 11+Myco MS-PREP®, Beer, Dietary exposure, Immunoaffinity columns, Mycotoxins, UPLC-MS/MS,
• MeSH
• chromatografie kapalinová metody   MeSH
• dietární expozice analýza   MeSH
• mykotoxiny * analýza   MeSH
• pivo analýza   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• mykotoxiny * MeSH

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.

• Klíčová slova
• HT-2 toxin, T-2 toxin, UPLC-MS/MS, immunoaffinity column, milk thistle, validation method,
• MeSH
• antioxidancia MeSH
• biologické přípravky * MeSH
• chromatografie kapalinová MeSH
• flavonoidy MeSH
• mykotoxiny * MeSH
• ostropestřec mariánský MeSH
• semena rostlinná MeSH
• silymarin * MeSH
• šlechtění rostlin MeSH
• T-2 toxin * analogy a deriváty   MeSH
• tandemová hmotnostní spektrometrie MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antioxidancia MeSH
• biologické přípravky * MeSH
• flavonoidy MeSH
• HT-2 toxin MeSH Prohlížeč
• mykotoxiny * MeSH
• silymarin * MeSH
• T-2 toxin * MeSH

Spices are a popular ingredient in cuisine worldwide but can pose a health risk as they are prone to fungal infestation and mycotoxin contamination. The purpose of this study was to evaluate ochratoxin A (OTA) in 54 single-kind traditional and less traditional spices, each of which was purchased in six samples of different batches (324 samples in total) at the Czech market during 2019-2020. The HPLC-FLD method with pre-treatment by immunoaffinity columns was employed to determine OTA. The limits of detection and quantification were 0.03 ng g-1 and 0.10 ng g-1, respectively. A total of 101 (31%) samples of 19 spice kinds were positive at concentrations ranging from 0.11-38.46 ng g-1. Only turmeric was contaminated with an OTA level exceeding the European Union limits. However, most spices have no regulation, thus further extensive monitoring of various mycotoxins in various kinds of spices is necessary. Chilli and black pepper are the most studied spices for OTA contamination, however, many other kinds of spice can also be highly contaminated, but studies on them are less common, rare, or have not yet been performed. The uniqueness of this study lies in the wide range of spice types studied for the presence of OTA on the Czech market.

• Klíčová slova
• HPLC-FLD, immunoaffinity columns, ochratoxin A, spices,
• Publikační typ
• časopisecké články MeSH

The European Union legislation regarding ochratoxin A (OTA) in various foodstuffs has changed relatively recently. Nevertheless, the legislation does not regulate OTA in any meat and meat-derived products. In this legislation update, the European Commission requested new studies, including, besides others, the presence of OTA in hams, which raises the concern that its consumption may pose a potential risk of exposure to OTA. This study aims to investigate OTA in a total of 195 samples of air-dry-cured hams acquired at the Czech market from January to June 2023. The analytical technique of high-performance liquid chromatography in combination with a fluorescence detector with pre-treatment employing immunoaffinity columns was used to determine OTA. OTA was found in 93 (48%) samples of air-dry-cured ham, with the OTA concentration reaching up to 14.58 ng/g. Due to the current absence of regulation limits, the results of this study were compared with the Italian maximum limit of 1 ng/g regulating OTA in porcine meat and byproducts. The Italian OTA maximum limit was exceeded in 22 (11%) samples. This study shows that the population of the Czech Republic is exposed to OTA from this pork byproduct. It is essential to set an OTA regulatory limit for meat and food produced from it to protect human health.

• Klíčová slova
• Food safety, HPLC-FD, Immunoaffinity columns, Mycotoxins, Nephrotoxicity,
• MeSH
• kontaminace potravin * analýza   MeSH
• masné výrobky * analýza   MeSH
• ochratoxiny * analýza   MeSH
• prasata MeSH
• vepřové maso analýza   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• ochratoxin A MeSH Prohlížeč
• ochratoxiny * MeSH

Carcinoembryonic cell adhesion molecule 1 (CEACAM1) is a human membrane glycoprotein belonging to the carcinoembryonic antigen (CEA) family and to the immunoglobulin superfamily. It is expressed in apical membranes of many epithelial cells in gastrointestinal and urogenital tract and also in granulocytes and lymphocytes, and its biological effect in human tissues has recently been discussed in literature. The purpose of this study was to isolate CEACAM1 glycoprotein from bile and characterize its purity and recovery which has not been described before. Affinity chromatography of CEACAM1 on hydrazide-activated cellulose with immobilized monoclonal anti-CEA F34-187 antibody is described. The immunoglobulin carbohydrate moiety was oxidized by periodate and then bound to hydrazide-activated matrix. Crude protein fraction from bile was applied on the affinity column and after extensive washing of non-bound proteins CEACAM1 was eluted with 6 M guanidine-HCl. A single immunopositive 85 kDa band was detected on Western blots with anti-CEA antibody after SDS-PAGE. We found out that CEACAM1 was not stainable with any common method of protein staining and the only non-specific method which could detect the 85 kDa band was a lectin staining.

• MeSH
• CD antigeny izolace a purifikace   MeSH
• celulosa chemie   MeSH
• chromatografie afinitní metody   MeSH
• diferenciační antigeny izolace a purifikace   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• hydraziny chemie   MeSH
• karcinoembryonální antigen imunologie   MeSH
• molekuly buněčné adheze MeSH
• monoklonální protilátky imunologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CD antigeny MeSH
• CD66 antigens MeSH Prohlížeč
• celulosa MeSH
• diferenciační antigeny MeSH
• hydraziny MeSH
• karcinoembryonální antigen MeSH
• molekuly buněčné adheze MeSH
• monoklonální protilátky MeSH

Mycotoxins are secondary metabolites of fungi and represent a serious problem for human health. Due to growing interest, various aspects have been widely studied by scientific groups. One of these aspects relates to the food industry and associated beer production. Mycotoxins can be present in the basic raw materials for beer production as well as in brewed beer. Problematic mycotoxins that pose a serious risk of toxicity are aflatoxins especially aflatoxin B1 (AFB1), fumonisins (FBs), and zearalenone (ZEN) and its metabolites, deoxynivalenol (DON) including its acetylated forms and also the modified form deoxynivalenol-3-glucoside (DON-3G), T-2 toxin, HT-2 toxin, and ochratoxin A. The Research Institute of Brewing and Malting has been dealing with the issue of mycotoxins since 2008. This study describes the analysis of the above mycotoxins during 2020-2024 in barley (n = 775), malt (n = 751), and commercially available beers (n = 522) using QuEChERS, immunoaffinity columns, and UPLC-MS/MS. The results showed positive samples of mycotoxins in brewing and malting matrices at the level of micrograms per kilogram (barley, malt) and nanograms per liter for beer. Therefore, it is a residual concentration and the accurate quantitative determination of mycotoxins, correct interpretation of the results in connection with toxicological values, and the maximum permissible levels of mycotoxins play a key role in global food safety and consumer protection.

• Klíčová slova
• Brewing, Immunoaffinity columns, Malting, Mycotoxins, QuEChERS, UPLC-MS/MS,
• MeSH
• ječmen (rod) chemie   mikrobiologie   MeSH
• jedlá semena * chemie   MeSH
• kontaminace potravin * analýza   MeSH
• lidé MeSH
• mykotoxiny * analýza   MeSH
• pivo * analýza   MeSH
• tandemová hmotnostní spektrometrie MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zearalenon analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• mykotoxiny * MeSH
• zearalenon MeSH

The co-occurrence of deoxynivalenol (DON) and deoxynivalenol-3-glucoside (DON-3-Glc) has been recently reported in malt and beer. In this study, the concentration changes were monitored within the brewing process of four beer brands: light, dark tap and two lagers, produced from ground malt mixtures differing in composition, and also mycotoxins content. A simple and rapid method employing DON-dedicated immunoaffinity columns (IAC) for the selective pre-concentration, followed by ultra-performance liquid chromatography coupled to a time-of-flight mass spectrometer (UPLC-TOFMS) system for the reliable quantification at (ultra)trace levels, was validated for all experimental matrices. The results document the key role of the malt contamination nature. While in the first monitoring period a significant increase of both DON and DON-3-Glc occurred (up to 250% and 450%, respectively), fairly different trends were observed when new malts were used for identical technological processing (in some beers a decrease of DON and only a small increase of DON-3-Glc occurred). Worth noticing, that the outcome of the brewing process was surprisingly reproducible for a particular malt mixture. In the final phase, a small monitoring study comparing Czech and Austrian alcohol-free and conventional beers was carried out.

• Klíčová slova
• Beer, DON-3-Glucoside, Immunoaffinity clean-up, Masked mycotoxins, Mycotoxins, UPLC–TOFMS, deoxynivalenol (DON),
• Publikační typ
• časopisecké články MeSH

An HPTLC method for the quantification of zearalenone (ZEA) in cereals and cereal products (wheat flour and malt) has been developed. ZEA was extracted with 50 ml of acetonitrile-purified water (9+1) with addition of 2 g NaCl. The extracts were further purified on VICAM ZearalaTest((tm)) immunoaffinity columns, then analysed by instrumental high-performance thin-layer chromatography (HPTLC) on silica gel plates with fluorescence detection. Ethyl acetate - n-hexane (1+1) was used as the mobile phase. The chromatogram was scanned in fluorescence mode after excitation at λ=254 nm with λ=400 nm measuring filter: SENS and SPAN parameters were 195 and 20, respectively. TheR F of ZEA under these conditions was 0.43.The recovery was 95% in the range 15-65 µg/kg cereal products; the mean relative standard deviation of repeatability (RSDr) was 7.6%. The limit of quantification (LoQ) of ZEA was 10 µg/kg. Validation of the method was performed according to the principles of the ICH for pharmaceutical analysis.

• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Medicinal plants are extensively utilized as dietary supplements to encourage disease prevention and to support the treatment of various health disorders. Unfortunately, several plants are known for mycotoxin contamination, which may overwhelm any beneficial effects the plants might have. OBJECTIVE: The purpose of the study was to determine the presence of ochratoxin A (OTA) and citrinin (CIT) in medicinal herbal products (MHP). METHODS: Sixty samples of different MHP types were purchased on the Czech market during 2020-2021. Both mycotoxins were determined using high-performance liquid chromatography with a fluorescence detector with immunoaffinity columns employed as a pretreatment. RESULTS: In total, 40% and 27% of samples were above the limit of quantification with the concentrations ranging up to 826.62 ng/g and 472.79 ng/g for OTA and CIT, respectively. The co-occurrence was confirmed in six MHP types. CONCLUSIONS: MHP could be a significant source of OTA and CIT. To protect the health of MHP users, it is desirable to continue monitoring the presence of mycotoxins in MHP. During this study, new OTA regulations for herbs came into force in the EU.

• Publikační typ
• časopisecké články MeSH

A new acrosin inhibitor was isolated to apparent homogeneity from the fluid of boar seminal vesicles. The inhibitor is immunologically related to the polyvalent trypsin-kallikrein inhibitor from bovine lung known as aprotinin. A crude preparation of the acrosin inhibitor was prepared by immunoaffinity chromatography on anti-aprotinin antibodies bound to Sepharose 4B column. The inhibitor was further purified by affinity chromatography on trypsin immobilized on a Sepharose 4B column, by ion-exchange chromatography on CM-Sephadex C-25, and by reversed-phase high-performance liquid chromatography on a C18 column. The relative molecular mass (Mr) of the inhibitor is about 7,000 as estimated from dodecyl sulfate-polyacrylamide gel electrophoresis. Its amino-acid composition was determined, the sequence of the first 8 amino-acid residues from the N-terminus is Thr-Arg-Asp-Phe-Pro-Pro-Asp-Gly-...

• MeSH
• akrosin antagonisté a inhibitory   imunologie   MeSH
• aminokyseliny analýza   MeSH
• chromatografie afinitní MeSH
• chromatografie iontoměničová MeSH
• inhibitory serinových proteinas * MeSH
• inhibitory trypsinu analýza   MeSH
• Kunitzův inhibitor trypsinu ze sójových bobů analýza   imunologie   MeSH
• molekulová hmotnost MeSH
• prasata MeSH
• semenné váčky analýza   MeSH
• skot MeSH
• tělesné tekutiny analýza   MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrosin MeSH
• aminokyseliny MeSH
• inhibitory serinových proteinas * MeSH
• inhibitory trypsinu MeSH
• Kunitzův inhibitor trypsinu ze sójových bobů MeSH