Chemometrics-Assisted Shotgun Proteomics for Establishment of Potential Peptide Markers of Non-Halal Pork (Sus scrofa) among Halal Beef and Chicken

Abstract

Authentication of meat is a global concern owing to meat adulteration with pork; however, no study, to date, has established a method to identify species-specific peptide markers for ascertaining adulterated meat. The present study aimed to describe chemometrics-assisted shotgun proteomics, which has recently been reported to establish potential peptide markers for non-Halal pork among Halal beef and chicken meat. To identify the potential peptide markers through peptide mass fingerprinting (PMF) analysis, chemometric analysis comprised principal component analysis and orthogonal partial least square-discriminant analysis, both of which proved statistically useful. Subsequently, through targeted tandem liquid chromatography-mass spectrometry (LC-MS) analysis, the primary structure of the identified peptide markers was revealed using the de novo identification approach. A decoy, randomised and concatenated database search program comprising MS-Fit and MS-Tag was used for PMF and targeted tandem LC-MS analysis, respectively. Results obtained using both PMF and targeted tandem LC-MS analysis complemented one another, indicating that these methods can yield consistent results to identify peptide markers together with chemometric analysis.