S. Maeda et al., Quantitative analysis of chondroitin sulfate isomers in intervertebral disk chondrocyte culture using capillary electrophoresis, J VET MED S, 63(9), 2001, pp. 1039-1043
Chondroitin sulfate (CS) isomers, 6-sulfate (CS6) and 4-sulfate CS4), chang
e their ratio to each other in cartilaginous tissues with aging. In this st
udy, a quantitative measurement method of CS6 and CS4 was developed, using
capillary electrophoresis (CE). Various buffer solutions, pH, and digestion
times were studied, and the use of 0.1 M Tris-HCl at pH of 8.0 allowed the
isolation of CS6 and CS4 from CS most efficiently when combined with chond
rotinase ABC at a concentration of 1 mU/mug of the substrate during a 3 hr
digestion period. Amounts of newly synthesized CS6 and CS4 in the intervert
ebral disk chondrocyte three-dimensional culture were quantified by this me
thod after the proteoglycans were extracted by equilibrium density centrifu
gation.