An Integrative Glycomic Approach for Quantitative Meat Species Profiling

Abstract

It is estimated that food fraud, where meat from different species is deceitfully labelled or contaminated, has cost the global food industry around USD 6.2 to USD 40 billion annually. To overcome this problem, novel and robust quantitative methods are needed to accurately characterise and profile meat samples. In this study, we use a glycomic approach for the profiling of meat from different species. This involves an O-glycan analysis using LC-MS qTOF, and an N-glycan analysis using a high-resolution non-targeted ultra-performance liquid chromatography-fluorescence-mass spectrometry (UPLC-FLR-MS) on chicken, pork, and beef meat samples. Our integrated glycomic approach reveals the distinct glycan profile of chicken, pork, and beef samples; glycosylation attributes such as fucosylation, sialylation, galactosylation, high mannose, α-galactose, Neu5Gc, and Neu5Ac are significantly different between meat from different species. The multi-attribute data consisting of the abundance of each O-glycan and N-glycan structure allows a clear separation between meat from different species through principal component analysis. Altogether, we have successfully demonstrated the use of a glycomics-based workflow to extract multi-attribute data from O-glycan and N-glycan analysis for meat profiling. This established glycoanalytical methodology could be extended to other high-value biotechnology industries for product authentication.

Analysis of Immunogenic Galactose-α-1,3-galactose-Containing N-Glycans in Beef, Mutton, and Pork Tenderloin by Combining Matrix-Assisted Laser Desorption/Ionization-Mass Spectroscopy and Capillary Electrophoresis Hyphenated with Mass Spectrometry via Electrospray Ionization

“…Of note, our off-tissue MALDI-TOF-MS data are supported by the MSI data which revealed relative abundances of α-Gal-containing N -glycans of approximately 50% in beef and 33% in pork. The possible explanation for the difference between our study and the report by Chia et al is the use of various analytical platforms (this study used MALDI-TOF-MS, whereas the reported data by Chia et al used LC–MS analysis) or/and construction of samples (tenderloin in this study versus shank cuts in the study by Chia et al).…”

“…Meat allergies induced by α-Gal following tick bites have only recently been appreciated as a world-wide problem . In-depth, meat glycomics are still limited and mainly based on liquid chromatography–mass spectroscopy (LC–MS) technologies. , Here, we present a comprehensive overview by analyzing the meat N -glycome using MALDI-TOF-MS and CE-ESI-MS, as exemplified by the identification of 53 and 55 N -glycans in beef and pork, respectively, versus the identified 18 and 17 N -glycans in a recent study by Chia et al . The similar points with the study by Chia et al are that we both present a similar level of Neu5Ac-containing N -glycans in beef and pork and a much higher level of Neu5Ac-containing N -glycans than Neu5Gc-containing N -glycans in pork (Supporting Information, Table S5).…”

“…While the role of α-Gal sensitization in mammalian meat allergy has thoroughly been investigated and demonstrated as illustrated above, the α-Gal moiety, as the terminal modification linked to N -glycans, has been rarely investigated in meat samples until now. Furthermore, protein glycosylation in meat samples is so far only studied using homogenized tissues. , Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) allows one to address this by generating two-dimensional illustrations of selected analytes by scanning the surface of biological samples (Figure b). − This methodology is generally used to detect analytes such as proteins, peptides, lipids, and metabolites, but various studies also report on its usage to explore the distribution of the N -glycome. − While MALDI-MSI has already been applied in food studies, , this technique has, to the best of our knowledge, not yet been exploited to investigate the N -glycome distribution of various meats. Therefore, the identity and spatial distribution of α-Gal - containing N -glycans in meat species is of keen interest from the nutritional perspective and consequently can provide scientific guidance for meat manufacturing, especially for processed meat products.…”

Detection of Authentication of Meat Products by Low Cost Closed-tube Molecular Methods

“…Meat authentication techniques are based on the identification of targeted analytes, namely biomarkers, or the identification of a chemical analyte pattern unique for the tested meat species. Classification of these techniques based on the group of the analytes to be detected includes DNA-based (classical techniques and genomics) (Dirong et al, 2021;Azad, 2023), RNA-based (transcriptomics) (Vishnuraj, Devatkal, Vaithiyanathan, Uday Kumar & Mendiratta, 2021), proteins-based (proteomics) (Stachniuk, Sumara, Montowska & Fornal, 2021;Suratno, Windarsih, Warmiko, Khasanah, Indrianingsih & Rohman, 2023), metabolites-based (metabolomics) (Zhang, Chen, Xie, Wang & Pan, 2021;Harlina, Maritha, Musfiroh, Huda, Sukri & Muchtaridi, 2022;Suratno, 2023), lipids-based (lipidomics) (Harlina et al, 2022;Jia, Di & Shi, 2023) and glycans-based (glycomics) based techniques (Shi et al, 2019;Chia et al, 2022).…”

Comprehensive Comparison of Bioactive N-Glycans among Seven Species of Livestock and Poultry Plasma Using a Relative Quantification Strategy

“…With rapid emergence of food glycomics as a new research area in recent years, the structure and functions of a series of bioactive glycans in many food materials have been revealed. , For instance, human milk is rich in diverse free oligosaccharides and glycoproteins, which not only have detoxification and anti-infection activities but also can be metabolized and utilized as natural prebiotics by intestinal flora, as well as the milk of many other mammals. − Specially, sialylated glycans in milk have proven to be essential to the development of infants’ brain learning functions. ,, Many other common foods, such as various eggs, many kinds of meats and some plant seeds, also contain a variety of glycoproteins, the glycans of which have important effects on human health. − Moreover, a few species of human milk bioactive glycans, such as 2′-fucosyllactose (2′-FL), have been commercially produced in industry and used as important functional additives for infant formula milk powder in recent years . However, the use of more kinds of healthful bioactive glycans in the food industry is hindered by the lack of large-quantity raw material resources for their production.…”