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The determination of mycotoxins in commercial beers
Martiník, Jan ; Benešová,, Karolína (referee) ; Mgr. Marek Pernica, Ph.D (advisor)
Mycotoxins are secondary metabolites of microscopic fibrous micromycetes that are able to infect cereals. The use of contaminated materials can lead to transfer of mycotoxins into the final product, such as beer. The master’s thesis deals with the determination of mycotoxins in beers. The theoretical part of this thesis describes selected mycotoxins, their occurrence, toxic properties and legislative limits of the European Union. The theoretical part also deals with the description of beer production and the possibilities of mycotoxin determination. The theoretical part also describes the statistical methods used for data processing. The experimental part of this work describes the validation of the method for determination of mycotoxins in beers. This section also describes the optimization of mycotoxin extraction using a commercially available 11+Myco MS-PREP® immunoaffinity column. The conditions for the determination of mycotoxins on UPLC-MS/MS are given in this thesis. The validation parameters such as linearity, accuracy, precision, LOD and LOQ were determined. This section contains a description of beer samples used for the determination of mycotoxins. The goal of the thesis was to optimize and validate the method for determination of mycotoxins in beers. From the validation parameters, it was found that this method is suitable for its intended purpose, namely for mycotoxins aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, fumonisin B1, fumonisin B2, fumonisin B3, ochratoxin A, ochratoxin B, zearalenone, -zearalenol, -zearalenol, -zearalanol, -zearalanol, deoxynivalenol, 3-acetyldeoxynivalenol, T-2 toxin and HT-2 toxin. The recoveries of this method ranged from 72,2 % to 100,0 %. The validated method was used for determination of above-mentioned mycotoxins in 89 beers. Of the total number of beers, 37 were produced in the Czech Republic and 52 in other European countries. Mycotoxins deoxynivalenol, T-2 toxin and HT-2 toxin were found in all beer samples. Common mycotoxins included fumonisin B1, ochratoxin A, -zearalenol and 3-acetyldeoxynivalenol. The mycotoxins aflatoxin B1, aflatoxin B2, aflatoxin G1, fumonisin B2, fumonisin B3, zearalenone and ochratoxin B were identified in less than 50 % of the samples. Mycotoxins aflatoxin G2, -zearalenol, -zearalanol and -zearalanol were not determined in any tested samples. The results of the analysis were subjected to statistical processing where the concentrations determined in Czech and European beers were compared. Principal component analysis and correlation analysis were created for aflatoxin B1, fumonisins, ochratoxin A, -zearalenol, deoxynivalenol, 3-acetlydeoxynivalenol, T-2 toxin and HT-2 toxin. The results of the analysis were compared with published studies.
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The determination of mycotoxins in commercial beers
Martiník, Jan ; Benešová,, Karolína (referee) ; Mgr. Marek Pernica, Ph.D (advisor)
Mycotoxins are secondary metabolites of microscopic fibrous micromycetes that are able to infect cereals. The use of contaminated materials can lead to transfer of mycotoxins into the final product, such as beer. The master’s thesis deals with the determination of mycotoxins in beers. The theoretical part of this thesis describes selected mycotoxins, their occurrence, toxic properties and legislative limits of the European Union. The theoretical part also deals with the description of beer production and the possibilities of mycotoxin determination. The theoretical part also describes the statistical methods used for data processing. The experimental part of this work describes the validation of the method for determination of mycotoxins in beers. This section also describes the optimization of mycotoxin extraction using a commercially available 11+Myco MS-PREP® immunoaffinity column. The conditions for the determination of mycotoxins on UPLC-MS/MS are given in this thesis. The validation parameters such as linearity, accuracy, precision, LOD and LOQ were determined. This section contains a description of beer samples used for the determination of mycotoxins. The goal of the thesis was to optimize and validate the method for determination of mycotoxins in beers. From the validation parameters, it was found that this method is suitable for its intended purpose, namely for mycotoxins aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, fumonisin B1, fumonisin B2, fumonisin B3, ochratoxin A, ochratoxin B, zearalenone, -zearalenol, -zearalenol, -zearalanol, -zearalanol, deoxynivalenol, 3-acetyldeoxynivalenol, T-2 toxin and HT-2 toxin. The recoveries of this method ranged from 72,2 % to 100,0 %. The validated method was used for determination of above-mentioned mycotoxins in 89 beers. Of the total number of beers, 37 were produced in the Czech Republic and 52 in other European countries. Mycotoxins deoxynivalenol, T-2 toxin and HT-2 toxin were found in all beer samples. Common mycotoxins included fumonisin B1, ochratoxin A, -zearalenol and 3-acetyldeoxynivalenol. The mycotoxins aflatoxin B1, aflatoxin B2, aflatoxin G1, fumonisin B2, fumonisin B3, zearalenone and ochratoxin B were identified in less than 50 % of the samples. Mycotoxins aflatoxin G2, -zearalenol, -zearalanol and -zearalanol were not determined in any tested samples. The results of the analysis were subjected to statistical processing where the concentrations determined in Czech and European beers were compared. Principal component analysis and correlation analysis were created for aflatoxin B1, fumonisins, ochratoxin A, -zearalenol, deoxynivalenol, 3-acetlydeoxynivalenol, T-2 toxin and HT-2 toxin. The results of the analysis were compared with published studies.
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