Comparison of heat-stable peptides using a multiple-reaction monitoring method to identify beef muscle tissue

Daniil Khvostov,Irina Chernukha,Natalya Vostrikova

doi:10.5219/1317

Abstract

Nowadays, proteomics is widely used as an analytical control method. A new method for determining animal tissue species-specificity based on a combination of two effective methods of food analysis, liquid chromatography (LC) and mass spectrometry (MS), was used in this work. Using this approach, it became possible to detect peptides. This work presents a comparison of species-specific, heat-stable peptides for the identification of beef. The objects of the study were native and boiled model mixtures containing beef with concentrations of 8% (w/w) and 16% (w/w). Pork was also added to the recipe to control for false-positive results. A high-performance liquid chromatography technique with mass spectrometric detection (LC-MS/MS) was used. Analysis of finished samples takes 25 minutes and is adapted to detect marker peptides. From the processing of the obtained data, three beef marker peptides were identified that were accepted as the best candidates. Two peptide prototypes, NDMAAQYK and YLEFISDAIIHVLHAK from the myoglobin protein and SNVSDAVAQSAR from the triosephosphate isomerase protein, were selected as potential biomarkers. For all samples, the signal-to-noise ratio (S/N) was set above 10. Temperature was not found to affect the structure and detection of marker peptides in samples with a muscle tissue concentration of 8% (w/w) at p <0.05. This approach is universally applicable for comparing biomarkers of other types of meat and to identify the most suitable candidates.

Highlights

Over the past 15 years, extensive research has been conducted around the world on the study of protein substances in raw meat and meat products, both native and those formed in the process of various technological treatments.A classic quantification method in proteomics is the use of an isotopic tag, the modification of which has more than 40 species (Kopylov and Zgoda, 2007)
A new method for determining animal tissue species-specificity based on a combination of two effective methods of food analysis, liquid chromatography (LC) and mass spectrometry (MS), was used in this work
In our work with the search for parameters for biomarkers on a mass spectrometer, the Skyline program proved to be the best. This is the best choice in the presence of a previously studied peptide sequence for develop of MRM methods

Summary

Introduction

A classic quantification method in proteomics is the use of an isotopic tag, the modification of which has more than 40 species (Kopylov and Zgoda, 2007). Various post-translational modifications of proteins with high sensitivity and specificity are studied by the Selected Reaction Monitoring (SRM) method (Zav'yalova, et al, 2014). A method of identifying species-specific molecular markers in the field of food analysis has gained strength, based on a combination of two methods, high-performance liquid chromatography (HPLC) and mass spectrometry (MS), used to detect peptides. Using this method, up to 0.5% (w/w) chicken meat was found in meat mixtures (Sentandreu et al, 2010). The good thermal stability of the peptides was demonstrated by the authors to identify horse and pork markers a lower limit of 0.24% (Von Bargen, Brockmeyer and Humpf, 2014)

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Conclusion