ARS | Publication request: ESTIMATION OF THE SULFATED GLYCOSAMINOGLYCAN CONTENT OF BOVINE SKIN WITH ALCIAN BLUE

Submitted to: Journal of American Leather Chemists Association
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 30, 2002
Publication Date: September 20, 2003
Citation: MOZERSKY, S.M., WILDERMUTH, R.J., MARMER, W.N. ESTIMATION OF THE SULFATED GLYCOSAMINOGLYCAN CONTENT OF BOVINE SKIN WITH ALCIAN BLUE. JOURNAL OF AMERICAN LEATHER CHEMISTS ASSOCIATION. 2003. V. 98. P. 337-343.

Interpretive Summary: Hides and skins consist of complex mixtures of interacting molecules. some of which undergo changes or removal during conversion into leather. It has been difficult to track some of these molecules to determine the effect of their removal or modification on the ultimate properties of leather. One target molecule is called decorin. Past attempts have not yielded conclusive evidence of its fate. Now, an analytical procedure has been perfected that uses a blue dye that coordinates with decorin by virtue of a sulfate handle on the target. By applying several steps that include enzyme digestion, dye application, centrifugation, and redissolving, the content of decorin may be correlated with the resultant blue color, which may be measured instrumentally. This new approach simplifies the assay substantially, and provides the leather researcher with a tool to determine the effect of the presence or absence of the target material on the quality of leather as well as to determine the adequacy of certain steps in the tanning process.

Technical Abstract: Procedures selected from the work of Bjornsson and Karlsson for the quantitation of sulfated glycosaminoglycans (SGAGs) with Alcian Blue were modified and adapted to the measurement of this type of polysaccharide in bovine skin. Modifications include: (1) pulverization of the skin under liquid nitrogen; (2) brief, gentle digestion of the skin with collagenase; (3) the use of a variety of Alcian Blue widely used in histologic work; (4) the recipe for the preparation of the Alcian Blue reagent solution; and (5) determination of suitable sample size, volumes of reagent solutions, incubation times, duration of centrifugation, etc. The procedure developed yields linear results for 50-300 mg samples of skin; the relative standard deviation for the procedure is ±15%.