ABSTRACT: Advanced Glycation End-Products (AGEs) are a heterogeneous group of compounds resulting from the non-enzymatic condensation between reducing sugars and amino groups, primarily lysine and arginine moieties (Perrone et al., 2020). The in vivo glycation of proteins is part of normal metabolism, but pathological implications, as well as molecular aging (e.g. protein conformation changes, inflammaging), have been associated with its occurrence (Briceno Noriega et al., 2022). In addition to the important deleterious effects of the endogenous formation of AGEs, the amount acquired from the diet is the subject of concern. In industrial or household cooking processes, the development of appealing aromas, flavors, and colors goes hand-in-hand with the formation of dietary AGEs (dAGEs) (Lund & Ray, 2017). It is well documented that thermally processed foods are a key dietary source of these neo-formed compounds, and high exposure to dAGEs has been associated with the pathogenesis of chronic non-communicable diseases (e.g. diabetes, obesity, and heart failure) (Clarke et al., 2016). Endogenous and dAGEs are thought to exert their pro-inflammatory and pro-oxidative effects at least in part through binding with the Receptor for Advanced Glycation End-products (RAGE) (Nogueira Silva Lima et al., 2021). Our goal here was to compare the efficiency of different reducing agents and carbonyl compounds for dCML fortification of bovine serum albumin (BSA) using a novel combination of quantitative and qualitative methods. Isotope-dilution, high-performance liquid chromatography with tandem mass spectrometric detection (HPLC-MS/MS), western-blot, fluorescence, and proteomics analyses using matrix-assisted laser desorption ionization with time-of-flight MS (MALDI-TOF) and LC with high-resolution MS (LC-HRMS) were all employed to provide an extensive characterization of the possible modifications of BSA promoted by different carbonyl compounds. We further studied the efficiency of the incorporation of protein-bound dCML into mouse feed pellets, their stability over different storage periods, as well as clearance of dCML in mouse feces.
BRAY FABRICE 
