Meat Authentication: A New HPLC–MS/MS Based Method for the Fast and Sensitive Detection of Horse and Pork in Highly Processed Food

Abstract

Fraudulent blending of food products with meat from undeclared species is a problem on a global scale, as exemplified by the European horse meat scandal in 2013. Routinely used methods such as ELISA and PCR can suffer from limited sensitivity or specificity when processed food samples are analyzed. In this study, we have developed an optimized method for the detection of horse and pork in different processed food matrices using MRM and MRM(3) detection of species-specific tryptic marker peptides. Identified marker peptides were sufficiently stable to resist thermal processing of different meat products and thus allow the sensitive and specific detection of pork or horse in processed food down to 0.24% in a beef matrix system. In addition, we were able to establish a rapid 2-min extraction protocol for the efficient protein extraction from processed food using high molar urea and thiourea buffers. Together, we present here the specific and sensitive detection of horse and pork meat in different processed food matrices using MRM-based detection of marker peptides. Notably, prefractionation of proteins using 2D-PAGE or off-gel fractionation is not necessary. The presented method is therefore easily applicable in analytical routine laboratories without dedicated proteomics background.

Authentication of processed meat products by peptidomic analysis using rapid ambient mass spectrometry

“…At present, sensitivity rises when chromatographic separation is involved, for example pork or horse meat can be detected down to 0.24% in processed products using HPLC-MS/MS method coupled with multiple reaction monitoring (von Bargen et al, 2014). In the aforementioned article, five peptide markers for processed pork and horse meat have been identified.…”

“…Recently, considerable improvement in mass spectrometry (MS) instrumentation has enabled the detection of peptide markers by liquid chromatography-MS techniques (LC-MS) and this has enabled identification of specific proteins from soybean (Leitner, Castro-Rubio, Marina, & Lindner, 2006), fish (Carrera et al, 2011) and meat species (Buckley et al, 2009;Montowska & Pospiech, 2013;Sentandreu, Fraser, Halket, Patel, & Bramley, 2010;von Bargen, Brockmeyer, & Humpf, 2014). In our previous work, we evaluated ambient MS techniques for standard protein identification in mixtures and for the analysis of meat digests to discriminate between five meat species (Montowska, Rao, Alexander, Tucker, & Barrett, 2014).…”

Authentication of pork in meat mixtures using PRM mass spectrometry of myosin peptides

“…For example, in the European "horse meat scandal" in 2013, at least 50 000 tons of beef meat contained horse meat and 5-7.5% of samples analyzed in the European Union (EU) contained undeclared horse meat. 3 Reliable analytical strategies for meat authentication are urgently needed to detect trace amounts of meat species. ELISA and PCR are two common methods used for species authentication.…”

Effect of animal species on muscle colour characteristics

“…Meat purchasing decisions are influenced more by product appearance than any other quality factor (Mancini, Hunt, 2005; Ngapo et al, 2007; Wu, Sun, 2013; Gracia, de Magistris, 2013). Consumers use colour as an intrinsic cue to predict experienced sensory quality (Banović et al, 2012) Although nowadays there are genetic methods for meat identification (Bargen et al, 2014;Fajardo et al, 2014; Santos et al, 2012), colour is one of the most important attributes that determine animal species identification and consumer acceptance of meat. The perception of colour is a very complex phenomenon that depends on the composition of the object in its illumination environment, the characteristics of the perceiving eye and brain, and the angles of illumination and viewing (Wu, Sun, 2013 (Tapp et al, 2011).…”