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Original
Determination of glucuronic acid containing fucoidan from liquid chromatography with pre-column fluorescence derivatization |
| Itsuko Kimura-Takagi, Masato Nagaoka*, Hideyuki Shibata, Shusuke Hashimoto, Sadao Ueyama, Ritsuo Aiyama and Teruo Yokokura |
Yakult Central Institute for Microbiological Research
1796 Yaho, Kunitachi-shi, Tokyo 186-8650, Japan |
A simple and highly sensitive high-performance liquid chromatographic method for the direct determination of glucuronic acid containing fucoidan from Cladosiphon okamuranus Tokida and related polysaccharides is described. The method is based on the direct derivatization of the uronic acid residues in polysaccharides with 6,7-dimethoxy-1-methyl-2 (1 H) -quinoxalinone-3-propionylcarboxylic acid hydrazide. The derivatization reaction proceeds in aqueous solution in the presence of pyridine and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide at 20 ℃. The resulting fluorescent derivatives are separated on reversed phase polymer column and are detected spectrofluorimetorically at 445 nm with excitation at 367 nm. The method is useful for the detection and quantitive analysis of uronic acid containing polysaccahrides.
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| Keywords: | fucoidan, uronic acid, polysaccharide, HPLC, DMEQ-hydrazide |
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