Metabolomic insights for the authentication and traceability of antibiotic-free pork: a pilot study.

BACKGROUND: Antibiotic-free production within the meat supply chain is a source of concern since new voluntary label claims need to be authenticated to prevent fraud. Therefore, an untargeted 1H NMR-based metabolomics approach coupled to unsupervised (principal component analysis) and supervised (orthogonal partial least squares discriminant analysis) multivariate analyses was applied to kidney (n = 47) and diaphragm (n = 54) samples of pigs from four Italian fattening farms to investigate changes in polar and nonpolar metabolite levels and differentiate between pigs treated with antibiotics (ABT) and pigs never exposed to antibiotic administration (ABF). RESULTS: The findings revealed a full discrimination between ABF and ABT pig samples, particularly when considering the aqueous extract of the investigated tissues. All discriminating metabolites differently accumulated in the polar extracts were annotated and grouped according to their molecular structure under the different biochemical categories (carbohydrates, amino acids and derivatives, organic acids, alcohols, choline and derivatives, and nucleosides) to discuss the putative biological function of the metabolism involved in response to antibiotic administration. CONCLUSIONS: Among the biochemical categories listed, amino acids and derivatives represented most (60%) of the affected metabolites in relation to antibiotic administration. © 2026 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.