Determination of the molecular weight distribution of non-enzymatic browning products formed by roasting of glucose and glycine and studies on their effects on NADPH-cytochrome c-reductase and glutathione-S-transferase in Caco-2 cells.

Abstract

After thermal treatment of a mixture of glucose and glycine for 2 h at 125 degrees C, about 60% of the starting material was converted into non-soluble, black pigments, whereas 40% of the mixture was still water-soluble. Dialysis of the latter fraction revealed 30.4% of low molecular weight compounds (LMWs; MW < 10,000 Da) and 10.0% high-molecular weight products (HMWs; MW > or = 10,000 Da). The water-soluble Maillard reaction products (MRPs) were separated by gel permeation chromatography and ultrafiltration, revealing that 60% of the water-soluble products of the total carbohydrate/amino acid mixture had MWs < 1,000 Da and consisted mainly of non-coloured reaction products. MRPs with MWs between 1,000 and 30,000 Da were found in comparatively low yields (about 1.3%). In contrast, about 31.1% of the MRPs exhibited MWs > 30,000 Da, amongst which 14.5% showed MWs > 100,000 Da, thus indicating an oligomerisation of LMWs to melanoidins under roasting conditions. To investigate the physiological effects of these MRPs, xenobiotic enzyme activities were analysed in intestinal Caco-2 cells. For Phase-I NADPH-cytochrome c-reductase, the activity in the presence of the LMW and HMW fraction was decreased by 13% and 22%, respectively. Phase-II glutathione-S-transferase activity decreased by 15% and 18%, respectively, after incubation with the LMW and the HMW fractions. Considering the different yields, 30% and 10%, respectively, of the LMW and the HMW fractions, the total amount of the LMW fraction present in the glucose-glycine mixture is more active in modulating these enzyme activities than that of the HMW fraction.