Quantitative analysis of chondroitin sulfate isomers in intervertebral disk chondrocyte culture using capillary electrophoresis

Citation
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
Citations number
23
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Veterinary Medicine/Animal Health
Journal title
JOURNAL OF VETERINARY MEDICAL SCIENCE
ISSN journal
0916-7250 → ACNP
Volume
63
Issue
9
Year of publication
2001
Pages
1039 - 1043
Database
ISI
SICI code
0916-7250(200109)63:9<1039:QAOCSI>2.0.ZU;2-9
Abstract
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.