Advanced glycation end products (AGEs) as pathology biomarkers: microwave assisted synthesis and biological quantitative analysis by LC-FTMS.

摘要

The Maillard reaction is a complex set of reactions, initially between carbonyl groups of reducing sugars and amino groups. Reactions of sugars with aminoacids or proteins are of great interest in food chemistry and in pathology. In fact AGEs accumulate in long lived tissue proteins in correlation with the development of complications in aging, diabetes, atherosclerosis and multiple sclerosis. Our research strategy has therefore focused on the quantification of specific AGEs biomarkers (pentosidine, glyceraldehyde derived pyridinium: GLAP) which could be associated with early pathology syndromes on rats fed high fat and fructose. In this work we synthesized pentosidine and GLAP by microwave assisted reactions in order to obtain standard compounds at improved yields and optimize the LC-FTMS quantitative analysis method. The analytical sensitivity of different LC detection methods was compared: HRMS~(2) (LTQ-Orbitrap), MS~(2) (triple quad), fluorescence detector (FD). The effect of different sample preparation methods (including soft and hard protein hydrolysis) was evaluated. The bioanalytical determination was finally applied to an animal study: glycation of proteins has been implicated in disease mechanisms. AGEs have a wide range of stabilities and varied biological effects. Here we described the effects of a high sugar diet given up 6 months to mice.