Detection of meat species adulteration using high-resolution mass spectrometry and a proteogenomics strategy

Due to the internationalisation of food production and distribution, there has been a significant increase of food fraud in recent years. Food fraud can have serious health implications, and it occurs when food manufacturers implement unethical practices such as making false label claims as well as using additives and fillers within their products to increase profitability. This has been a serious concern. Meat adulteration was examined using a well-defined proteogenomic annotation, carefully selected surrogate tryptic peptides and high-resolution mass spectrometry. Selected mammalian meat samples were homogenised and the proteins extracted and digested with trypsin. Chromatography was achieved using a 30-min linear gradient along with a BioBasic C8 100 × 1 mm column at a flow rate of 75 µl min^-1. The mass spectrometer was operated in full-scan high-resolution and accurate mass using resolving powers of 140,000 and 17,500 (FWHM) in full-scan MS and MS/MS respectively. Data independent acquisition (DIA) mode was used including 12 DIA MS/MS scans to cover the mass range 600-1200 m/z. Methodically in silico analyses of myoglobin, myosin-1, myosin-2 and β-haemoglobin sequences allow for the identification of a species-specific tryptic peptide mass lists and theoretical MS/MS spectra. Following comprehensive MS, MS/MS or DIA analyses, the method was capable of the detection and identification of very specific tryptic peptides for all four targeted proteins for each animal species tested with observed m/z below 3 ppm compared with the theoretical m/z. The analyses were successfully performed with raw and cooked meat. Specifically, the method was capable of detecting 1% (w/w) of pork or horse meat in a mixture before and after cooking (71°C internal temperature).