# Untargeted lipidomics profiling for halal authentication of meatball products from mixed meat sources

## TL;DR

This study uses lipidomics and advanced data analysis to detect halal and non-halal meat in meatballs, even when meats are mixed.

## Contribution

The study introduces untargeted lipidomics as a novel method for halal authentication in processed meat products.

## Key findings

- Lipid profiles from beef, goat, and pork were clearly differentiated using UHPLC–HRMS and chemometrics.
- PCA and PLS-DA models showed high accuracy in separating halal and non-halal meatball samples.
- Five and twelve lipid markers were identified for single-source and mixed-source meat products, respectively.

## Abstract

Food adulteration with non-halal components, particularly in thermally processed meat products like meatballs, presents significant challenges for conventional DNA and protein-based detection methods. This study applies untargeted lipidomics using ultra-high-performance liquid chromatography–high-resolution mass spectrometry (UHPLC–HRMS) combined with multivariate analysis (principal component analysis [PCA] and partial least squares-discriminant analysis [PLS-DA]) to authenticate halal status in meatball products. Lipid profiles were obtained from beef, goat, pork, and their mixtures, and analyzed under both electrospray ionization in positive mode (ESI+) and electrospray ionization in negative mode (ESI−) modes. Glycerophospholipids emerged as dominant lipid species. PCA and PLS-DA revealed clear separations between halal and non-halal samples, with high model performance (R2 = 0.994, Q2 = 0.952), demonstrating strong predictive capability. Five key lipid ions—lysophosphatidylethanolamine (LPE)(16:0p) + H, LPE(16:1e) + H, LPE(18:1p) + H, PC(10:0p/23:0) + H, and phosphatidylcholine (PC)(8:0/24:1) + H—were identified as markers for single-source differentiation, while twelve lipids were significant for mixed-meat products. These findings confirm the efficacy and specificity of untargeted lipidomics as a robust method for halal authentication in processed meat products.

•Lipidomics and chemometrics were used to authenticate halal meatball products.•UHPLC–HRMS successfully detected distinct lipid profiles in single and mixed meat sources (beef, goat, pork).•PCA and PLS-DA identified specific lipid biomarkers to discriminate halal and non-halal meatball compositions.•Five lipid markers identified for single-source and twelve for mixed-source products.

Lipidomics and chemometrics were used to authenticate halal meatball products.

UHPLC–HRMS successfully detected distinct lipid profiles in single and mixed meat sources (beef, goat, pork).

PCA and PLS-DA identified specific lipid biomarkers to discriminate halal and non-halal meatball compositions.

Five lipid markers identified for single-source and twelve for mixed-source products.