Heat-stable peptide markers specific to rabbit and chicken liver tissue for meat product authentication testing

A three-step analysis was used to detect and identify heat-stable peptide markers specific to liver tissue from rabbit and chicken. It involved peptide discovery by liquid chromatography coupled to high resolution mass spectrometer (LC-HRMS), followed by protein identification using Spectrum Mill software and multiple reaction monitoring (MRM) based confirmation of the discovered peptides using a liquid chromatography coupled to triple quadrupole mass spectrometer (LC-TQ). We identified 50 and 91 heat-stable peptide markers unique to chicken and rabbit liver, respectively. The markers were validated in commercial food samples with declared liver tissue contents ranging from 5% to 30%. The best candidate peptides for distinguishing liver tissue from skeletal muscle were selected and then confirmed using MRM-based approach. Limit of detection of liver was found to be in the range of 0.13 to 2.13% (w/w) for chicken liver-specific peptide markers, and from 0.04 to 0.6% (w/w) for rabbit liver-specific peptide markers.