The Measurement of Tissue Acid Mucopolysaccharides1

Abstract

Dried, defatted tissue was extracted in 0.5N NaOH. The extract was neutralized with acetic acid and then perchloric acid was added to precipitate protein. After dialysis against water, aliquots were precipitated with protamine, and uronic acid determined by carbazole and orcinol methods. Good reproducibility and satisfactory recovery of chondroitin sulfate was found. Human dermis, subcutaneous tissue, aorta and thyroid tissue were analyzed; aorta had the highest concentration of acid mucopolysaccharide, thyroid the second highest. A portion of the extract of each tissue was digested by testicular hyaluronidase. Chromatography of extracts of each tissue revealed three components, with mobilities resembling hyaluronic acid, chondroitin sulfate A and heparin. The ratio of carbazole to orcinol values for dermis was 0.76, falling to 0.52 after hyaluronidase incubation, suggesting the presence of chondroitin sulfate B. The ratio of carbazole to orcinol values for aorta averaged 1.5, suggesting the presence of heparitin sulfate.