Chemiluminescence high-performance liquid chromatography for the determination of hyaluronic acid, chondroitin sulphate and dermatan sulphate.
A sensitive chemiluminescence high-performance liquid chromatographic method has been developed for the determination of hyaluronic acid, chondroitin sulphate and dermatan sulphate as their unsaturated disaccharide-dansylhydrazine derivatives involving an effective sample clean-up system. The dansylhydrazones of the unsaturated disaccharides derived from the hyaluronic acid, chondroitin sulphate and dermatan sulphate by chondroitinase ABC and/or chondroitinase ACII, were separated by reversed-phase chromatography using a mixture of 0.1 M sodium acetate buffer (pH 6.0) and 80% acetonitrile on a column (250 mm x 4.0 mm I.D.) packed with amide-80 silica beads (5 microns diameter). For post-column elution in the chemiluminescence system, 1 mM bis[2-(3,6,9-trioxadecanyloxycarbonyl)-4-nitrophenyl]oxalate and 3mM hydrogen peroxide in acetonitrile were used. The detection limit of each glycosaminoglycan was 100 fmol. The method was applicable to the determination of the levels of hyaluronic acid, chondroitin sulphate and dermatan sulphate in rat peritoneal mast cells.
Akiyama
H
H
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Shidawara
S
S
Mada
A
A
Toyoda
H
H
Toida
T
T
Imanari
T
T
D000818:Animals; D002809:Chondroitin Sulfates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003871:Dermatan Sulfate / Q000032:analysis*; D006820:Hyaluronic Acid / Q000032:analysis*; D008163:Luminescent Measurements; D008407:Mast Cells / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar
D003871:Dermatan Sulfate; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates