Preliminary report on fractionation of fucans by ion-exchange displacement centrifugal partition chromatography |
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Affiliation: | 1. Research Institute, JCR Pharmaceuticals Co., Ltd., Kobe 651-2241, Japan;2. R&D Platform Technology and Science, GlaxoSmithKline, King of Prussia, Pennsylvania 19406;3. R&D Platform Technology and Science, GlaxoSmithKline, Stevenage, UK |
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Abstract: | A new method combining ion-exchange displacement chromatography with centrifugal partition chromatography (CPC) was used for the fractionation of partially depolymerized fucans (polysulphated polysaccharides). The ion-exchanger was Amberlite LA2, a high-molecular-mass liquid secondary amine miscible with most common organic solvents and immiscible with aqueous solutions. Ion-exchange displacement centrifugal partition chromatography was performed with LA2 in methyl isobutyl ketone (MiBK) as the stationary phase, water as the mobile phase, Cl− as the carrier and OH− as the displacer. A complex mixture of partially depolymerized fucans was resolved into adjacent families characterized by their peak molecular mass and polydispersity. The Dubois test (sugar) and the azur A test (SO3−) confirmed the displacement mode of the process, and size-exclusion chromatographic controls confirmed its efficiency. |
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