The study was carried out in an alpine area of North-Eastern Italy to assess the reliability of proton nuclear magnetic resonance 1H NMR to fingerprint and discriminate Asiago PDO cheeses processed in the same dairy plant from upland pasture-based milk or from upland hay-based milk. Six experimental types of Asiago cheese were made from raw milk considering 2 cows’ feeding systems (pasture- vs. hay-based milk) and 3 ripening times (2 months, Pressato vs. 4 months, Allevo_4 vs. 6 months, Allevo_6). Samples (n = 55) were submitted to chemical analysis and to 1HNMRcoupled with multivariate canonical discriminant analysis. Choline, 2,3-butanediol, lysine, tyrosine, and some signals of sugar-like compounds were suggested as the main water-soluble metabolites useful to discriminate cheese according to cows’ feeding system. A wider pool of polar biomarkers explained the variation due to ripening time. The validation procedure based on a predictive set suggested that 1H NMR based metabolomics was an effective fingerprinting tool to identify pasture-based cheese samples with the shortest ripening period (Pressato). The classification to the actual feeding system of more aged cheese samples was less accurate likely due to their chemical and biochemical changes induced by a prolonged maturation process.
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