Analysis of dog meat adulteration in beef meatballs using non-targeted UHPLC-Orbitrap HRMS metabolomics and chemometrics for halal authentication study

Due to the different price and high quality, halal meat such as beef can be adulterated with non-halal meat with low price to get an economical price. The objective of this research was to develop an analytical method for halal authentication testing of beef meatballs (BM) from dog meat (DM) using a non-targeted metabolomics approach employing liquid chromatography-high-resolution mass spectrometry (LC-HRMS) and chemometrics. The differentiation of authentic BM from that adulterated with DM was successfully performed using partial least square-discriminant analysis (PLS-DA) with high accuracy ((RX)-X-2 = 0.980, and (RY)-Y-2 = 0.980) and good predictivity (Q(2) = 0.517). In addition, partial least square (PLS) and orthogonal PLS (OPLS) were successfully used to predict the DM added (% w/w) in BM with high accuracy (R-2 > 0.990). A number of metabolites, potential for biomarker candidates, were identified to differentiate BM and that adulterated with DM. It showed that the combination of a non-targeted LC-HRMS Orbitrap metabolomics and chemometrics could detect up to 0.1% w/w of DM adulteration. The developed method was successfully applied for analysis of commercial meatball samples (n = 28). Moreover, pathway analysis revealed that beta-alanine, histidine, and ether lipid metabolism were significantly affected by dog meat adulteration. In summary, this developed method has great potential to be developed and used as an alternative method for analysis of non-halal meats in halal meat products.

Automated summary

This research developed an analytical method using liquid chromatography-high-resolution mass spectrometry and chemometrics to successfully differentiate beef meatballs from those adulterated with dog meat. The combination of non-targeted metabolomics and chemometrics can detect up to 0.1% of dog meat adulteration. The study also identified the significant impact of dog meat adulteration on beta-alanine, histidine, and ether lipid metabolism. This method has the potential to serve as an alternative method for analyzing non-halal meats in halal meat products.