New Sensitive High-Performance Liquid Chromatography–Tandem Mass Spectrometry Method for the Detection of Horse and Pork in Halal Beef

Abstract

The accidental or fraudulent blending of meat from different species is a highly relevant aspect for food product quality control, especially for consumers with ethical concerns against species, such as horse or pork. In this study, we present a sensitive mass spectrometrical approach for the detection of trace contaminations of horse meat and pork and demonstrate the specificity of the identified biomarker peptides against chicken, lamb, and beef. Biomarker peptides were identified by a shotgun proteomic approach using tryptic digests of protein extracts and were verified by the analysis of 21 different meat samples from the 5 species included in this study. For the most sensitive peptides, a multiple reaction monitoring (MRM) method was developed that allows for the detection of 0.55% horse or pork in a beef matrix. To enhance sensitivity, we applied MRM(3) experiments and were able to detect down to 0.13% pork contamination in beef. To the best of our knowledge, we present here the first rapid and sensitive mass spectrometrical method for the detection of horse and pork by use of MRM and MRM(3).

Proteomics in Meat Science — Current Status and Future Perspective

“…The possi-время изучается возможность применения протеом-ного подхода для обеспечения идентификации видов мяса, но данные все еще ограниченные. Von Bargen et al [20] разработал чувствительный масс-спектроме-трический метод с мониторингом множественных ре-акций (MRM), который позволил определить низкие концентрации свинины и конины в образцах халяль-ной говядины.…”

“…Позже Marco-Ramell et al [26] идентифицировали глутатион пероксидазу, гли-копротеин α2 Heremans Schmid, холестерин и фекаль-ный кортикостерон как потенциальные биомаркеры bility to apply a proteomic approach to enable identification of meat species is under investigations, but data is still limited. Von Bargen et al [20] developed sensitive multiple reaction monitoring (MRM) mass spectrometry method which allowed to detect low concentrations of pork and horse meat in halal beef samples.…”

Advances in Authenticity Testing for Meat Speciation

“…The use of LC-MS/MS with a multiple reaction monitoring (MRM) method for the identification of specific peptides was also proposed for the identification of horse and pork in halal beef (von Bargen et al, 2013) and in highly processed foods (von Bargen et al, 2014). The method allowed detection of 0.55% (w/w) of horse or pork in beef, or down to 0.13% pork in beef when MRM 3 and a micro-LC system were applied (von Bargen et al, 2013).…”

“…The method allowed detection of 0.55% (w/w) of horse or pork in beef, or down to 0.13% pork in beef when MRM 3 and a micro-LC system were applied (von Bargen et al, 2013). Since the selected marker peptides were proven to be sufficiently stable to heat processing, the same methodology was further used for the identification of pork and horse meat in highly processed foods (von Bargen et al, 2014).…”

Assessment of meat authenticity using bioinformatics, targeted peptide biomarkers and high-resolution mass spectrometry

“…Recently a study was published using a shotgun bottom-up proteomic approach using peptide mass fingerprinting (PMF) and MS to determine meat authenticity (Sentandreu & Sentandreu 2011;von Bargen et al 2013). There are limitations with the PMF approach proposed, including data analysis and pattern complexity that frequently reduce assay sensitivity to determine meat authenticity accurately.…”