Extraction, Isolation and Analysis of Chondroitin Sulfate Glycosaminoglycans

Abstract

Glycosaminoglycans (GAGs) including chondroitin sulfate (CS) and chondroitin sulfate/dermatan sulfate (CS/DS) copolymers are anionic straight chain polysaccharides. They are galactosamine containing GAGs (galactosaminoglycans) having wide range of applications in pharmaceutical, cosmetic and food industries. This article reviews techniques to isolate and characterize these galactosaminoglycans from animal and poultry tissues. Patent based information is also discussed. Cartilaginous tissues are the major source of CS consisting entirely of D-glucuronosyl-N-acetylgalactosamine repeating disaccharide units, in which the galactosamine is sulfated at C4 or C6. In contrast, most galactosaminoglycans in non-cartilaginous connective tissues (e.g. skin and tendon) are CS/DS copolymers comprised of varying proportions of D-glucuronosyl-N-acetylgalactosamine and L-iduronosyl-N-acetylgalactosamine. Tissues are digested with proteinase (e.g. papain) to liberate GAGs, which are fractionated to isolate and purify galactosaminoglycans. Common techniques used for fractionation of GAGs include: precipitation with different concentrations of ethanol; solubilization of GAG precipitated as GAG-quarternary ammonium compound complexes with different concentrations of NaCl; anion exchange chromatography and gel filtration chromatography. Purified galactosaminoglycans are examined by various methods including chondroitinase digestion, high performance liquid chromatography and electrophoresis. Histological methods are used to localize galactosaminoglycans in tissues. The patent information on the CS hydrolase and ultraviolet irradiation may be useful for the preparation of CS oligosaccharide.

Established method of chondroitin sulphate extraction from buffalo (Bubalus bubalis) cartilages and its identification by FTIR

“…However, in the present study after electrophoresis the gel was visualised by staining with coomassie brilliant blue. Nakano et al ( 2010 ) studied the extraction, isolation and analysis of CS glycosaminoglycan and determined CS molecular mass by polyacrylamide gel electrophoresis and also reported that the mobility of CS was depending on its charge density.…”

“…There was no significant difference in CS electrophoresis pattern extracted from different sources. However, Nakano et al ( 2010 ) reported that PAGE has been used to estimate molecular mass of glucosaminoglycans but, it is not much sensitive to detect it’s presence in small variation in the molecular mass, and the results of electrophoresis of CS samples had shown similar migration pattern of standard CS. This indicates that non CS proteins are effectively removed during the extraction process.…”

Chondroitin sulphate extracted from antler cartilage using high hydrostatic pressure and enzymatic hydrolysis

“…The aim of this work was to determine the effect of high pressure, temperature and incubation time on the catalytic activity of papain for extracting CS as a potential antioxidant agent. GAGs can be directly extracted from tissues by hydrolysis with exogenous enzymes like papain or pronase [20]. Further separation of the CS in the crude extract was obtained by column chromatography, and the hyaluronic acid binding ability of CS was also examined.…”