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Three methods for determination of patulin in apple juice have been studied comparatively. The selected procedure for liquid samples implies liquid-liquid extraction with ethyl acetate/hexane (96:4, v/v). The collected patulin was then analyzed by liquid chromatography with UV-Visible detection. The obtained recovery was 82.9±3.2 % and the limit of detection was 2.0 µg/kg of juice, which are according to the valid guidelines established by the European Union. The developed analytical method has been used for the determination of patulin in various Spanish apple juices, where this mycotoxin has not been detected.