Determination of Meat Authenticity Using a Comprehensive Targeted Proteomic Strategy and High-Resolution Mass Spectrometry

Importance of the Topic

Protein based methods for verifying meat authenticity are crucial due to rising food fraud and its implications for public health and regulatory compliance.

Objectives and Study Overview

• Develop a targeted proteomic workflow to confirm meat species identity
• Use high resolution Orbitrap mass spectrometry to detect species specific peptides
• Assess sensitivity of method using meat mixtures containing adulterant levels down to 1 percent

Methodology

• Prepare raw beef horse lamb and pork mixtures at defined weight ratios from 100 down to 1 percent
• Extract proteins and perform tryptic digestion for 24 hours at 40C
• Separate peptides by reversed phase liquid chromatography on a C8 column with formic acid and acetonitrile mobile phases
• Acquire full scan MS data over m z 500 to 2000 at 140000 resolution
• Perform targeted MS MS at 17500 resolution with collision energy set to 25 and 1.5 Da isolation
• Process data using Thermo Scientific Proteome Discoverer software for peptide identification

Instrumentation Used

• Thermo Scientific Q Exactive quadrupole Orbitrap mass spectrometer
• Thermo Scientific Dionex UltiMate 3000 RSLC system
• Thermo Scientific BioBasic C8 column 100 x 1 mm 5 micron

Main Results and Discussion

• Four proteotypic myoglobin peptides spanning positions 120 to 134 provided clear species discrimination with mass accuracy below 1.3 ppm
• Additional peptides from myosin 1 myosin 2 and beta haemoglobin reinforced species specificity
• Extracted ion chromatograms demonstrated reliable detection of 1 percent adulterant in complex mixtures
• Both precursor and fragment ion monitoring delivered consistent performance for meat speciation

Advantages and Practical Applications

• High sensitivity allows detection of undeclared species at low levels
• Robust targeted assay suitable for routine quality control in food testing laboratories
• Comprehensive peptide panel reduces false positives and enhances confidence in results

Future Trends and Potential Applications

• Incorporate sample enrichment techniques to push detection limits below 1 percent
• Extend the approach to additional species and processed food matrices
• Adopt automated data independent acquisition for broader peptide monitoring
• Integrate rapid sample preparation for high throughput screening

Conclusion

Targeted high resolution proteomics using Orbitrap mass spectrometry offers a reliable strategy to confirm meat species identity and detect minor adulteration down to 1 percent with potential for further sensitivity improvements.

References

1. Alberto Ruiz Orduna Erik Husby Charles T Yang Dipankar Ghosh Francis Beaudry Assessment of meat authenticity using bioinformatics targeted peptide biomarkers and high resolution mass spectrometry Food Additives Contaminants Part A 2015 DOI 10.1080/19440049.2015.1064173