A specific analytical method has been developed for the simultaneous determination of some mycotoxins in vegetable oils: Aflatoxins (AF), alpha-zearalenol (alpha-ZEA), zearalenone (Zea), ochratoxin A (OTA) were extracted and purified in a single step by SPE on polymeric cartridge (PPL) and quantified by RP-HPLC with fluorescence detection. The proposed method, validated for accuracy and precision parameters, shows reliable results matching criteria suggested by the European Union for Mycotoxins Analysis.