A method for the selective release and, hence, assay of the carbohydrate moieties of collagen

The carbohydrate residues of collagen were selectively released in high yield by nitrosation of the hydroxylysines of the intact collagen or peptides derived from collagen. The carbohydrate residues (Glc-Gal and Gal) released were separated from the modified protein or peptide by gel chromatography and were assayed by gas-liquid chromatography of their trimethylsilyl derivatives. The results agreed closely with those obtained from methanolysis-gas chromatography or from alkaline hydrolysis followed by amino acid analysis of the hydroxylysyl glycosides. With a more sensitive perbenzoylation-high-performance liquid chromatography method and uv detection at 230 nm, the carbohydrates released by nitrosation of submilligram quantities of collagen or peptide could be assayed accurately.