Advanced glycation and oxidation products

Advanced glycation end products (AGEs) are formed during nonenzymatic reactions of reducing sugars and other reactive carbonyls with proteins.

Together with glycation, proteins in vivo are also modified by reactive oxygen species (ROS). These reactions are also accelerated in diabetes and end stage renal disease.

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Protein
modifications by
Vitamin C

Viatmin C oxidation products play a role in the browning process of lens crystallins.

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Instrumentation for AGEs quantification

 

Absolute quantification of advanced glycation end products (AGEs) in proteins requires either total protein hydrolysis with hydrochloric acid at 100 ºC for 16 h for acid stable AGEs (CML, CEL, argpyrimidine, pentosidine), or extensive enzymatic digestion by a combination of different peptidases for all AGEs.
After chromatographic separation fluorescent compounds such as argpyrimidine and pentosidine can be detected by fluorescence detector connected to HPLC, however, most AGEs are not fluorescent and application of mass spectrometry for their quantification by isotope dilution with selected ion monitoring gas chromatography/mass spectrometry (SIM-GC-MS) or multiple reaction monitoring liquid chromatography tandem mass spectrometry (MRM-LC-MS/MS) is required.
SIM-GC-MS requires amino acids and AGEs derivatization into their N,O-trifluoroacetyl methyl ester (TFAME) derivatives.
Our laboratory provides analytical techniques to measure AGEs via LC-MS/MS and/or GC/MS in biological samples. Request a quote.