Zearalenone is a mycotoxin produced by some Fusarium species in food and feed. Toxicity of ZEA and its metabolites is related to the chemical structure of the mycotoxins, similar to naturally occurring estrogens. Currently, there is increasing awareness of the significance of fungi and their toxic metabolites in water. Thus it was considered essential to determine the concentration of estrogenic toxin in surface waters, groundwater and wastewater in Poland. The application of ZearalaTest immunoaffinity columns for zearalenone determination by HPLC method in water samples characterized by different natural organic matter (NOM) content gave the recovery rate ranging from 74 to 86%. Obtained results showed that the natural organic micropollutants of water with low molecular masses have an effect on zearalenone recovery. Zearalenone was present in water samples in the range from 0 to 43.7 ngL(-1).