Fate of the Amino Acid in Glucose−Glycine Melanoidins Investigated by Solid-State Nuclear Magnetic Resonance (NMR)

Abstract

The fate of the amino acid in the model Maillard reaction between glucose and glycine in a 1:1 molar ratio has been investigated by applying advanced 13C and 15N solid-state nuclear magnetic resonance (NMR) techniques to 13C- and 15N-labeled melanoidins formed in dry and solution reactions. Quantitative 13C NMR shows that approximately 23% of carbon is from glycine; the approximately 2% loss compared to the 25% glycine C in the reactants is due to the COO moiety being liberated as CO2 (Strecker degradation). 13C J-modulation experiments on melanoidins made from doubly 13C-labeled glycine show that the C-C backbone bond of about two-thirds of the incorporated amino acid stays intact, and about half of all glycine is incorporated as N-CH2-COO without fragmentation. Degradation processes without CO2 loss affect about one-eighth of glycine in dry reaction and about one-fourth in solution. These results indicate that Strecker degradation affects about one-fourth (dry reaction) to one-third (in solution) of all glycine but is not the main pathway of glycine incorporation. Spectra of Strecker degradation products show that C2 of glycine reacts to form N-CH3, C-CHn-C, or aromatic units, but not pyrazines or pyridines. The gycine-C1 carbon incorporated into the melanoidins remains>or=90% part of COO moieties; approximately 5% of amides have also been detected. The C2-N bond stays intact for approximately 70% of the incorporated glycine. The 15N spectra show many peaks, over a 200 ppm range, documenting a multitude of different chemical environments of nitrogen, but no enamines or imines. The majority (>78%) of nitrogen, in particular most pyrrolic N, is not protonated. Because N-H predominates in amino acids and proteins, nonprotonated nitrogen may be a characteristic marker of Maillard reaction products.