Chromatographic analysis of hydroxylysine glycosides and acid hydrolyzates

A single-column chromatographic technique for the analysis of hydroxylysine glycosides and acid hydrolyzates is described. This technique employs a Durrum D-500 amino acid analyzer equipped with a standard 48 cm X 1.8 mm column packed with DC-4A resin. Resolution was achieved with four sodium citrate buffers and four column temperatures. Products of glycoprotein hydrolysis including cysteic acid, methionine sulfoxide, 4-hydroxyproline, alpha-aminobutyric acid, glucosamine, galactosamine, hydroxylysine, tryptophan and the internal standard, norleucine, are resolved. The completely automated procedure takes 185 min per run and can measure amino acid residues in the nanomole range.