title: Simultaneous determination of uronates found in polysaccharides from natural products by HPLC with fluorometric detection.
abstract: A sensitive high-performance liquid chromatographic (HPLC) method for the determination of uronates isolated from polysaccharides found in natural products such as glycosaminoglycans and alginate is described. Preparation of iduronate, guluronate, and mannuronate as analytical standards for high performance liquid chromatography was achieved by depolymerization of dermatan sulfate and alginate in 2.5 mol L(-1) trifluoroacetic acid at 100°C for 6h. Structures of resulting products (iduronate, guluronate, and mannuronate) were characterized by 600 MHz (1)H NMR. Five uronates (glucuronate, iduronate, mannuronate, guluronate, and galacturonate) were separated on a Dionex CarboPac PA1 column using an isocratic elution with 8 mmol L(-1) acetate buffer (pH 4.84) and were monitored by fluorescence detection using 1.5% 2-cyanoacetamide as a post-column fluorogenic reagent. As little as 50 pmol of each uronate could be detected with excitation at 331 nm and emission at 383 nm.
authors: Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan.
authors: Akihisa
authors: A
authors: Matsumoto
authors: Saori
authors: S
authors: Hosoyama
authors: Kyohei
authors: K
authors: Higashi
authors: Toshihiko
authors: T
authors: Toida
mesh_terms: D000464:Alginates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003871:Dermatan Sulfate / Q000737:chemistry; D005470:Fluorometry / Q000379:methods*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D007067:Iduronic Acid / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D013696:Temperature; D014269:Trifluoroacetic Acid / Q000737:chemistry; D014574:Uronic Acids / Q000737:chemistry*
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D011134:Polysaccharides; D014574:Uronic Acids; C007896:guluronic acid; D003871:Dermatan Sulfate; D007067:Iduronic Acid; D020723:Glucuronic Acid; C008324:mannuronic acid; D014269:Trifluoroacetic Acid