The biogenic amine content of “Provola dei Nebrodi”, a typical “pasta filata” Sicilian cheese, was studied at different ripening stages. Reversed-phase high performance liquid chromatography (RP-HPLC) with fluorimetric detection was employed for the identification and quantification of eight different amines: ethanolamine, histamine, serotonine, tyramine, tryptamine, 2-phenylethylamine, putrescine and cadaverine. Histamine was the most representative amine in all the analysed cheese samples, followed by serotonin and tyramine. The total amount of biogenic amines was lower than 0.1 ppm in fresh samples, whereas it reached 33.57 ppm in samples with the longest ripening time. The biogenic amine contents were correlated with the proteolytic maturation coefficient, defined as the water-soluble nitrogen/total nitrogen (WSN/TN %) percent ratio. The results showed amine contents lower than the toxic level even for samples with the longest ripening time.