A reliable method for the simultaneous determination of aflatoxins B1, G1, B2, G2, zearalenone and its metabolites α-zearalanol and β-zearalanol was developed. Mycotoxins have been extracted from eggs using a QuEChERS-based extraction procedure (quick, easy, cheap, effective, rugged and safe) followed by applying C18 and a PSA further clean-up step, then detection by ultra high performance liquid chromatography (UPLC) coupled to tandem mass spectrometry (MS/MS) using an electrospray–ionization interface (ESI) in both positive and negative ion modes. Matrix-matched calibration was used for quantification in order to reduce the matrix effect. Validation of the method was carried out in eggs by recovery experiments. Recoveries of the spiked samples were in the range between 84.8 and 114.6% with an intra-day relative standard deviation lower than 20.7% and an inter-day relative standard deviation lower than 18.2%. Limits of quantification (LOQ) ranged from 3.0–15.0 μg kg−1. The developed method was successfully applied to the analysis of eggs and a positive sample which was contaminated by AFB1 was detected.
