Application of metabolomics and chemometrics to detect buffalo meat adulteration in beef meatball emulsion products

Species substitution, most notably substituting buffalo for beef, makes it difficult to ensure that meat quality is authentic and halal. In this study, 600 MHz ¹H NMR metabolomics and chemometrics were employed for the detection of buffalo meat adulteration in a meatball containing beef. The composition of beef and buffalo meat was mixed with emulsion to prepare meatballs at a 99:1, 95:5, 90:10, and 80:20 (w/w) ratio. Metabolomic profiling of the untargeted metabolites was carried out, which revealed 45 metabolites including significant changes in amino acid, purine catabolism, and glycolysis metabolic pathway, with an increase in buffalo content leading to a significant decrease in most of free amino acid and energy related metabolites; taurine and ATP levels increased as a function of buffalo content. Supervised orthogonal partial least square-discriminant analysis models were created to achieve high discrimination rates at substitution of beef with buffalo content of more than 10% with Q² > 0.876, while ATP, inosine, and one unidentified but high discriminatory factor were found as the most discriminating markers. The results revealed that the NMR-based metabolomics approach is a non-destructive approach for verifying the meat authenticity in the heat-treated food and a potential tool for the halal monitoring industry in the world.