Quantitation and interaction of glycosaminoglycans with Alcian blue in dimethyl sulfoxide solutions. |
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Authors: | L Hronowski T P Anastassiades |
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Affiliation: | 2. Department of Medicine and Biochemistry, Rheumatic Diseases Unit,Queen''s University, Kingston, Ontario K7L 3J7 Canada |
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Abstract: | An improved method is described for the quantitation of glycosaminoglycans separatedon cellulose acetate, stained with Alcian blue, and dissolved in a dimethyl sulfoxide solution. Standard curves are shown for all eight glycosaminoglycans. It is shown that absorption at the Alcian blue orthochromatic Emax is depressed under conditions which favor formation of dye-glycosaminoglycan complexes. The interaction between Alcian blue and the eight glycosaminoglycans was studied in dimethyl sulfoxide solutions of varying composition. It was shown that the extent of complex formation depends both on the glycosaminoglycan and the composition of the dimethyl sulfoxide solution. A dimethyl sulfoxide solution which contains 0.094 m H2SO4 is described which maximizes dye-glycosaminoglycan dissociation and thus the absorbance. Also, an improved staining method is described which improves dye uptake by the glycosaminoglycans and consequently increases the sensitivity of glycosaminoglycan quantitation. |
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Keywords: | Requests for reprints should be addressed to Dr. T. Anastassiades Department of Medicine Queen's University 26 Barrie Street Kingston Ontario K7L 3J7 Canada. |
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