A method has been obtained for the analysis of hexosamines and neutral sugars by gas-liquid chromatography. Hexuronic acids were found not to be resolved by the standard method and needed to be reduced under acid conditions. As this procedure was shown to be unreliable, polysaccharides containing both hexuronic acids and hexosamines were subjected to such modification reactions as methylation, but with no results. Hydrolytic procedures were applied to glycosaminoglycans to obtain the maximal yields of hexosamine and hexuronic acid. The same procedures were applied to samples of costal cartilage powder and, in addition, conditions for the maximum release of hydroxyproline, sialic acid, calcium and magnesium were determined. The normal range for each of the cartilage constituents was determined and the hydrolysis conditions applied to known cases of metabolic bone disease. In each case an abnormality in at least two constituents was observed. These abnormalities suggest areas for more intensive research and this simple approach to cartilage chemistry is regarded as a 'screening' procedure. Colourimetric hexuronic acid analysis of cartilage was non-reproducible and a new method involving modification of the acid moieties in glycosaminoglycans is presented.
|Date of Award||1976|