title: Assessment of meat authenticity using bioinformatics, targeted peptide biomarkers and high-resolution mass spectrometry.
abstract: In recent years a significant increase of food fraud has been observed, ranging from false label claims to the use of additives and fillers to increase profitability. Recently in 2013 horse and pig DNAs were detected in beef products sold from several retailers. Mass spectrometry (MS) has become the workhorse in protein research, and the detection of marker proteins could serve for both animal species and tissue authentication. Meat species authenticity is performed in this paper using a well-defined proteogenomic annotation, carefully chosen surrogate tryptic peptides and analysis using a hybrid quadrupole-Orbitrap MS. Selected mammalian meat samples were homogenised and proteins were extracted and digested with trypsin. The samples were analysed using a high-resolution MS. 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 MS was operated in full-scan high resolution and accurate mass. MS/MS spectra were collected for selected proteotypic peptides. Muscular proteins were methodically analysed in silico in order to generate tryptic peptide mass lists and theoretical MS/MS spectra. Following a comprehensive bottom-up proteomic analysis, we detected and identified a proteotypic myoglobin tryptic peptide (120-134) for each species with observed m/z below 1.3 ppm compared with theoretical values. Moreover, proteotypic peptides from myosin-1, myosin-2 and β-haemoglobin were also identified. This targeted method allowed comprehensive meat speciation down to 1% (w/w) of undesired product.
authors.affiliation: a Département de biomédecine vétérinaire, Faculté de médecine vétérinaire , Université de Montréal , Saint-Hyacinthe , QC , Canada.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ruiz Orduna
authors.affiliation: b Department of Environment & Food Safety , Thermo Fisher Scientific , San Jose , CA , USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Husby
authors.affiliation: b Department of Environment & Food Safety , Thermo Fisher Scientific , San Jose , CA , USA.
authors.forename: Charles T
authors.initials: CT
authors.lastname: Yang
authors.affiliation: b Department of Environment & Food Safety , Thermo Fisher Scientific , San Jose , CA , USA.
authors.forename: Dipankar
authors.initials: D
authors.lastname: Ghosh
authors.affiliation: a Département de biomédecine vétérinaire, Faculté de médecine vétérinaire , Université de Montréal , Saint-Hyacinthe , QC , Canada.
authors.forename: Francis
authors.initials: F
authors.lastname: Beaudry
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D005504:Food Analysis / Q000295:instrumentation / Q000379:methods*; D005506:Food Contamination / Q000032:analysis* / Q000517:prevention & control; D059022:Food Safety; D006736:Horses; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D008460:Meat / Q000032:analysis*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis*; D010449:Peptide Mapping; D059748:Proteolysis; D016415:Sequence Alignment; D013552:Swine; D014357:Trypsin / Q000737:chemistry
chemical_list: D015415:Biomarkers; D009124:Muscle Proteins; D010446:Peptide Fragments; D014357:Trypsin
keywords: HPLC; authenticity; biomarkers; food; high-resolution mass spectrometry; meat; proteomics