Stable isotope dilution analysis of the Fusarium mycotoxin zearalenone

Zearalenone is a secondary metabolite produced by molds of the Fusarium genus. Beside its nonsteroidal molecular structure, zearalenone has estrogenic activity and can disrupt the function of the endogenous hormone 17 ss-estradiol in animals and possibly in humans. It can frequently be found in all major cereal grains as well as in processed food. Because of the estrogenic properties of zearalenone and its metabolites, legal regulations are installed in the European Union setting maximum levels in cereals and cereal products. Routine analysis of zearalenone in various commodities is carried out by HPLC with fluorescence detection, but due to the development of multi-mycotoxin methods and the reduced sample cleanup, HPLC-MS/MS has become a fast and efficient alternative. However, to achieve a reliable quantitation with this technique suitable internal standards are required. This paper reports the synthesis of stable isotope labeled 3,5-d(2)-zearalenone (ZON) as internal standard for stable isotope dilution analysis. Furthermore, a method for the analysis of zearalenone by HPLC-MS/MS using 3,5-d2-zearalenone as IS has been developed. Fifteen cereal products from the German retail markets were analyzed, of which seven contained ZON in levels from 4.9 to 45.0 mu g/kg.