Analysis of glycated amino acids by high-performance liquid chromatography of phenylthiocarbamyl derivatives.
A method has been developed for the analysis of hexitolamino acids formed by acid-catalyzed hydrolysis of nonenzymatically glycated proteins that have been treated with sodium borohydride. The hexitolamino acids are converted into phenylthiocarbamyl (PTC) derivatives which are analyzed by reverse-phase HPLC. The PTC derivatives of N alpha-hexitolamino acids behave like lactones, migrating on the column more slowly than the corresponding PTC-amino acids. The PTC derivatives of N epsilon-glucitol- and N epsilon-mannitol-lysine are probably free acids, since they migrate faster than PTC-lysine. The method, which can be used to determine the degree of glycation of N-terminal and lysyl residues, has been applied successfully to human hemoglobin, serum albumin, and ocular lens proteins.
Walton
D J
DJ
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
McPherson
J D
JD
D000596:Amino Acids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D017879:Isothiocyanates; D011506:Proteins / Q000032:analysis; D013861:Thiocyanates
D000596:Amino Acids; D017879:Isothiocyanates; D011506:Proteins; D013861:Thiocyanates; C005441:phenylisothiocyanate