A microdetermination method for assaying glycosaminoglycans and proteoglycans.

A simple and reliable method is described for the determination of glycosaminoglycans and proteoglycans in tissue extracts as well as during preparative and analytical procedures for these molecules. It is particularly useful because it requires much less starting material for the identification of glycosaminoglycans or proteoglycans and, in addition, is several fold more sensitive than the currently used uronic acid assays. The procedure allows separation of macromolecules by ion-exchange chromatography, density gradient centrifugation, or molecular sieve chromatography and involves spotting onto cellulose acetate membrane, reaction with Alcian blue, and quantitation of color in a spectrophotometer. This method is particularly appropriate to use for the analysis of proteoglycans and glycosaminoglycans in tissues which are available in limited amounts or have low levels of these macromolecules.