NON-ENZYMIC GLYCATION OF PROTEINS: ANALYSIS OF N-(1-DEOXYHEXITOL-1-YL)AMINO ACIDS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Article abstract of DOI:10.1016/S0008-6215(00)90270-2

A method for determining the extent of non-enzymic glycation (originally called "glycosylation") of both lysyl and N-terminal residues of a protein is described.The glycated protein is treated with sodium borohydride, and is then subjected to acid-catalysed hydrolysis.The resulting N-(1-deoxy-D-hexitol-1-yl)amino acids are separated by cation-exchange high-performance liquid chromatography (l.c.), and detected by a post-column reaction with periodate.The method has been applied successfully to samples of human hemoglobin and human serum albumin, for measurement of numbers of valine-attached and of lysine-attached N-(1-deoxy-D-fructos-1-yl) groups per protein molecule.