The volatile concentrate obtained from Grana Padano cheese by vacuum distillation was fractionated by continuous liquid–liquid extraction into neutral and acid fractions. Both were analysed by high resolution gas chromatography (HRGC), HRGC–mass spectrometry, and HRGC–olfactometry. A total of 67 components were identified in the neutral extract (22 esters, 13 alcohols, 12 ketones, 6 aldehydes, 5 nitrogen-containing compounds, 3 lactones and 6 miscellaneous compounds) and 16 in the acid extract. Esters were the predominant constituents of the neutral fraction, whose major components were ethyl butanoate and ethyl hexanoate. HRGC–olfactometry of the neutral compounds demonstrated that 23 were odour-active: ethyl butanoate, 2-heptanol, 3-methylthiopropanal, 1-octen-3-ol, ethyl hexanoate and nonanal being the most potent odorants. n-Butanoic and n-hexanoic acids were the main volatile free fatty acids identified in the acid extract as having an important odour with a high olfactometric index. The backbone of Grana Padano cheese aroma seemed to consist of these acids and 14 potent neutral odorants imparting fruity, green, nutty and coconut notes. The concentration of volatile components responsible for the fruity and green notes was inversely proportional to the length of ripening, whereas the concentration of volatile agents with spicy, nutty and earthy notes tended to increase during maturation. In a comparison of the olfactometric profile, the Grana Padano cheese aroma was found to be more complex than an imitation Grana Padano cheese produced with similar technology but outside the area of the genuine cheese. Some of the main metabolic pathways for the biosynthesis of cheese aroma are reviewed briefly to indicate the possible origin of the compounds identified.