COUNTERCURRENT CHROMATOGRAPHIC SEPARATION OF SUGARS AND THEIR <i>p</i>-NITROPHENYL DERIVATIVES BY CROSS-AXIS COIL PLANET CENTRIFUGE

Abstract

Abstract Sugars and their p-nitrophenyl (PNP) derivatives were separated by high-speed countercurrent chromatography using a cross-axis coil planet centrifuge (cross-axis CPC) equipped with a pair of eccentric coil assemblies. A polar two-phase solvent system composed of 1-butanol/acetic acid/water (4:1:5) was used for the separation of sucrose and fucose, and glucuronic acid (lactoid form) and galacturonic acid, while 1-butanol/ethanol/water (4:1:4) was used for the separation of free and lactoid forms of glucuronic acid. PNP-sugar derivatives such as neutral sugars, uronic acids, and amino sugars were separated with a less polar solvent system composed of n-hexane/ethyl acetate/1-butanol/methanol/water at various volume ratios. PNP-glucose derivatives were further separated according to the number of sugar chains, and five PNP-neutral sugars were resolved by adding 0.1 M sodium tetraborate in the two-phase solvent system. Overall results of experiments revealed that the cross-axis CPC is useful for the separation of polar compounds such as sugars. ACKNOWLEDGMENT This work was supported in part by a grant from Nihon University, Tokyo, Japan.