Simultaneous determination of uronates found in polysaccharides from natural products by HPLC with fluorometric detection
| Autor: | Kyohei Higashi, Toshihiko Toida, Akihisa Matsumoto, Saori Hosoyama |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Glucuronate
Alginates Iduronic Acid Dermatan Sulfate Polysaccharide Biochemistry High-performance liquid chromatography Dermatan sulfate Analytical Chemistry chemistry.chemical_compound Glucuronic Acid Polysaccharides Trifluoroacetic acid Trifluoroacetic Acid Fluorometry Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography Depolymerization Hexuronic Acids Organic Chemistry Temperature General Medicine Uronic Acids chemistry Reagent Proton NMR |
| Zdroj: | Carbohydrate research. 358 |
| ISSN: | 1873-426X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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